Author Archives: author

A Pilot Study: Can Using QR Codes Increase Student Participation in Large Classroom Settings?

DOI: 10.31038/IJNM.2023432

Abstract

In large classes, students report feeling a lack of personalization and ownership for learning course material. Students can feel disengaged from the learning process, thus a greater risk of failing to seek clarification of misunderstood concepts. Given that 35-50% of persons have an introverted temperament, it is important to provide an equitable learning environment to support all students’ learning. The purpose of this pilot project was to determine if the use of a QR code to ask questions would improve student engagement both during class and after class time for a first and third semester prelicensure nursing students. Post QR code survey using a 1-10 Like rt scale showed a mean satisfaction score of 8.63 for 1st semester and 7.79 for 3rd semester students. 90.7% of first semester and 76.9% of third semester students felt having access to the QR code enabled them to ask more questions.

Keywords

QR code, Large class size, Student participation, Prelicensure nursing program, Engagement

Introduction

Nursing school faculty face many challenges supporting student participation in the classroom. One challenge is identifying appropriate technologies which will enhance student participation without burdening faculty and being perceived as useless by students. This can be particularly difficult in large classes. Students in large classes have reported feeling a lack of personalization and ownership for learning course material, disengagement from the learning process, and greater risk of failing to seek clarification of misunderstood concepts [1,2]. It has been noted the use of electronic devices can be distracting and discouraged [3]. However, the use of technology can promote critical thinking and knowledge retention [4]. Quick response (QR) codes were developed over 20 years ago and are widely used in business [5]. QR code readers are readily available on portable devices and have been incorporated in patient and medical education programs. Today, QR code readers are readily available on portable devices such cell phones and tablets. QR codes have been incorporated in patient and medical education programs, but there have been few applications of QR codes in prelicensure nursing programs [5-7]. The purpose of this paper is to describe the outcome of a pilot study at an accelerated baccalaureate nursing program (ABSN) in the Southeastern United States that applied QR code technology to improve student participation in a large classroom.

Background

Created in 1994, a QR code is a bar code which when scanned, links to a predetermined digital destination. Originally designed to improve inventory efficacy, QR codes are now part of everyday life. The use of QR codes has been studied, however, only relative to enhancing student experiences in low fidelity simulations4. By using QR codes, students were able to access video and audio assessment findings. Students felt the use of a QR code enabled them to pull together assessment findings and apply to nursing care. Focusing on specific breath sounds was one innovative example where QR codes have been useful in nursing education. By using a QR code, students were able to hear and document findings in an appropriate manner [8,9]. Finally, QR codes have been used to promote Just-in-time (JIT) learning for nursing skills and understanding sexually transmitted infections (STIs) [3]. After an extensive literature search, no research was found highlighting the use of a QR to ask questions either in class or outside of class.

Ensuring all students have a voice in the classroom is essential, particularly in large class sizes. This can be especially difficult for a student who is introverted. Introversion has been defined as persons who prefer quieter, less stimulating environments. The percentage of people who fall somewhere on an introverted temperament continuum is estimated to be 35% to 50%10. This can have a significant impact on student learning and stress. Calling on an introvert student to answer questions can add to the already high stress level. By recognizing the introversion-extroversion continuum, educators can create an equitable learning environment which support all students’ learning [10].

Method

This project received expedited Institutional Review Board approval. A comprehensive literature review searching for QR code usage in nursing education found no articles discussing the use of QR codes for large classroom participants. Based on this search, a pilot project was developed to implement QR codes in first and third semester courses for undergraduate students. To determine if there was a difference between beginning students and more advanced students. The authors developed a Redcap survey to develop baseline data. Redcap pre/post surveys were kept separate though both had the same questions. The four question pre-QR code survey asked students about experience in using a QR code, comfort in raising one’s hand in class to ask a question and if uncomfortable raising one’s hand, how were questions answered. To determine if there was a generational difference, an age range question was added. At semester’s end, a post-survey asked if students used the QR code to ask a question in class, used it outside of class, whether there was comfort in using the QR code, what was the satisfaction in using the QR code and did it lead to asking more questions. To implement the project, a QR code linked to a Redcap survey was developed. When the QR code was scanned, the student was taken to a Redcap survey, allowing the student to choose which faculty they wanted to respond to their question and then type a question. The question was immediately sent via email to the requested faculty so the faculty could respond in “real-time” during a class lecture. Students had the option to provide one’s name so faculty could respond personally or remain anonymous. The QR code was also imbedded in each power point/Panopto recording so students could ask a question outside of the classroom. The QR code was printed and laminated so each student could have it available during class time.

Results

All students from both cohorts completed the pre-survey and greater than ninety-eight percent had experience using QR codes with no statistical difference between the groups. What was surprising was the percentage of students who stated they were uncomfortable raising their hand in class which was approximately 28% from both groups. Comments for first semester students were: “Sometimes I am shy.” and “I feel comfortable in an online format asking questions but not in person classes.” Comments from third semester students included: “Yes, social anxiety prohibits me from asking questions routinely in class, especially when we’re all together again in one room!” and I will ask a question out loud, if need be, but I’d rather not.” An interesting finding was how students got questions answered. First semester students had a significantly higher dependence on course faculty either meeting after class or emails compared with third semester students who relied more on classmates for assistance. Over the course of the semester, first semester students used the QR code forty-one times while third semester students used the QR code eighteen times. While the project was designed to enable students to ask questions during class, greater than ninety-five percent of questions received were after class. For both cohorts, nearly all students chose to be anonymous. Responses to anonymous QR code questions were posted in the course announcement section of the learning management system (LMS) under the assumption if one student had a question, so would other students. Post QR code survey results showed a mean satisfaction score of 8.63 for first semester students on a 0 -10 scale with 10 being the highest and 7.79 for third semester students. Students were asked if they felt they could ask more questions due to the use of a QR code. For both semester groups, the results were very positive at 90.7% and 76.9% for first and third semester students respectively.

Discussion

This project demonstrated the feasibility of using QR codes to improve student participation in large classroom settings. The greatest benefit appeared to be with first semester students. This finding was not unexpected as first semester students may lack confidence or have anxiety about asking questions in a large classroom9. However, using the QR code enabled all students to ask questions throughout the semester. This approach provided immediate feedback to faculty on areas where students struggled. Several lessons were learned during the pilot study. One was remembering to place the QR code on student desks. This was adjusted for the next semester by creating a QR code which attached to the student’s badge. Since the QR code survey was anonymous, faculty could not provide a personal response. This was corrected by adding a statement to the QR code Redcap survey asking the student to provide a name if they wanted a personal email. Since the QR question went instantly to the faculty email, there was some difficulty in how frequently faculty checked emails during class. In one instance, there were several similar questions, but class time did not allow for an in-depth explanation. However, a detailed recorded explanation was posted after class.

Conclusion

Providing students with QR codes to anonymously ask questions both during and after class has the potential to promote student learning and participation by fostering an inclusive and responsive in a large classroom setting. In addition, it may decrease first semester anxiety as students adjust to a new learning environment as well as help instructors pinpoint where students are struggling with new concepts.

Conflict of Interest

The authors declare no conflict of interest

References

  1. Kinyon K, D’Alton S, Poston K, Anderson B (2021) Changing student outcomes without sacrificing student satisfaction. Journal of Nursing Education 60: 1-3. [crossref]
  2. Robb MK (2012) Managing a large class environment: Simple teaching strategies for new nurse educators. Teaching and Learning in Nursing 7: 47-50.
  3. Sommerall WE, Roche CC (2020) The ABC’s of STIs: Promoting student learning using QR codes. Journal of Nursing Education 59. [crossref]
  4. Marcus J, Adeboyo M, Cranwell-Bruce L, Faulkhner MS (2021) Evaluating nursing students’ perceptions of using quick response codes to enhance learning during nursing health assessment. Nursing Education Perspectives 42: E133-134. [crossref]
  5. Billing DM, Kowalski K, Bradley K (2020) Just-in-time learning and QR codes: A must-have tool for nursing professional development specialists. J Contin Educ in Nurs 51: 302-303. [crossref]
  6. Bellot J, Shaffer k, Wang M (2015) Use of quick response coding to create interactive patient and provider resources. Journal of Nurs Education. 54: 224-227. [crossref]
  7. Jacoby S (2012) QR-codes-Quick response. Radiologic Technology 83: 301-302.
  8. Marcus J, Adeboyo M, Cranwell-Bruce L, Faulkhner MS (2021) Evaluating nursing students’ perceptions of using quick response codes to enhance learning during nursing. Nursing Education Perspectives 42: E133-E134.
  9. Willbanks C (2021) Take out your phones! Using QR codes in a health assessment course to overcome social-distancing obstacles. Tennessee Nurse 19.
  10. Colley SL (2018) Equitable education for introverts: providing learning opportunities that prepare nursing students to be leaders. Nursing Educator 45: 244-226.

Comparison Analysis of Metabolites by Exercise in Thoroughbred and Korean Native Jeju Pony

DOI: 10.31038/EDMJ.2023723

Abstract

Objective: Among experimental animal models, horses are the most adaptable to exercise and this ability has been extensively studied. Research on equine exercise physiology is mostly focused on genetics, and few integrated studies have focused on equine metabolomics. This study were conducted to analyze metabolites in plasma, urine, and sweat samples collected from Jeju pony and thoroughbred horses before and after exercise. In this study, we analyze the various equine samples using NMR (nuclear magnetic resonance) spectroscopy.

Methods: 1H NMR spectroscopy analysis were conducted with equine plasma, urine, and sweat samples collected from Jeju pony and thoroughbred horses before and after exercise. Relative metabolite levels between three types of were compared under exercise stimuli and by breeds.

Results: A total 26, 39, and 36 metabolites were identified in each of plasma, sweat, and urine samples, respectively, of both thoroughbred and Jeju pony. A total 3, 12, 15 metabolites were exclusively detected in plasma, sweat, and urine samples, respectively, and 15 metabolites were detected in all samples at the same time. In addition, total 8 and 5 metabolites were detected after exercise in plasma and urine samples. Additionally, we obtained 16, 6, and 30 metabolites in plasma, urine, and sweat by breeds.

Keywords

Horse, Thoroughbred, Korean native horse, Jeju pony, Metabolites, Nuclear magnetic resonance spectroscopy

Introduction

Horses are the most adaptable experimental animal models to exercise and, as such, are the most suitable for studying its effects. Moreover, studies focused on exercise physiology in horses can provide valuable basic information for understanding underlying mechanisms associated with exercise in humans. For this reason, further research on exercise physiology is necessary [1]. However, although studies focused on improving the athletic performance of horses have not had much success, economic trait -related genes have recently received greater attention [2-4]. In addition, equine tissue derived cells are being used in studies on the functional validation of these genes [5,6].

In recent years, multivariate analyses, so called multi-omics (genomics, epigenomics, transcriptomics, metabolomics, and proteomics) have been used to explain the biological mechanisms in numerous animals. Metabolites are the final biological products of cellular processes in cells, tissues, organs, or organisms [7]. Quantification of metabolomes can explain several biological phenomena along with other omics studies.

Exercise has a powerful effect on the body metabolism [8]. Repetitive and unilateral contraction of muscles associated with frequent exercise training is a suitably strong stimulus of physiologic function. During exercise, blood-borne glucose, creatine phosphate, glycogen, free fatty acids, and lactate which is known as external molecular substrates were used to produce ATP in muscle. The importance of these external molecular substrates in exercise metabolism is mostly affected by exercise rate and duration, but can also be affected by the type of exercise itself, as well as diet and environmental factors [9]. In addition, other energy mechanisms may be needed depending on the degree and duration of exercise [10].

In a previous study, we investigated a metabolic mechanism activated by physical activity using 1H nuclear magnetic resonance (NMR) spectroscopy in thoroughbred horses [11]. Specifically, we profiled exercise specific metabolome in muscles and plasma. However, the metabolic mechanism during exercise has not yet been analyzed in urine and sweat, which are much easier to collect than plasma and muscle.

In this study, the metabolite profiling of the sweat, plasma, and urine in various equine breeds under exercise stimulus was analyzed by 1H NMR spectroscopy. Based on the results, commonly or specifically released metabolites were identified from various equine biopsied specimen. Subsequently, metabolic pathways associated with obtained metabolites were investigated. The present study could contribute to a better understanding of metabolic fluctuations caused by exercise in thoroughbred and Jeju pony.

Materials and Methods

Animals

In this study, samples were gathered from five Thoroughbred and five Jeju pony. The study design was approved by the Pusan National University-Institutional Animal Care and Use Committee (Approval Number: PNU-2015-0864).

Horse Sampling

Jeju pony and Thoroughbred horse samples of sweat, plasma, and urine were collected in a stable setting and following exercise (30 min). A 15 mL syringe was used to obtain blood samples, which were then transferred to heparin tubes and centrifuged at 5,000 rpm for 15 min to extract the plasma. Sweat samples were obtained only after exercise. In case of urine, obtained sample was centrifuged to remove solids. Supernatant of centrifuged urine samples was added to a 1.5 ml tube which, is containing D2O (deuterium oxide) solution, DSS( dextran sulphate sodium), and 10 mM imidazole. In addition, 0.42% sodium azide was added. Obtained plasma samples and sweat samples were stored at -20°C, and urine samples were stored at -70°C until conducting NMR spectroscopy.

Nuclear Magnetic Resonance Spectroscopy

Plasma, urine, and sweat samples were 1H NMR spectroscopy analyzed. Briefly, plasma, urine, and sweat samples were used with D2O containing the reference material TSP (trimethylsilylpropionate) before NMR measurement. We conducted high-resolution magic angle spinning NMR for plasma samples, with a spinning rate of 2,050 Hz. Water peak and macromolecular peak signals were removed using the Carr-Purcell-Meiboom-Gill pulse sequence for analysis of plasma, sweat, and urine samples. Used to eliminate signals from water peaks and macromolecular peaks. Measured spectrum data were optimized by Chenomx NMR Suite 7.1 (Chenomx Inc., Edmonton, AB, Canada), and statistical analysis were conducted by SIMCAp+12.0 (Umetrics, Umea, Sweden) software. In this study, we measured the absolute concentrations of the metabolites in various equine samples. Relative concentrations were determine, and amount of metabolites present in the samples were calculated by multivariate statistical analysis method.

Statistical Analysis

A T-test and One-way ANOVA analysis of variance was conducted to determine significance levels. Data were shown as mean ± standard deviation of mean. One-way ANOVA  analysis of variance followed by Duncan multiple test was used to compare before and after exercise training results and used for each sample of thoroughbreds and Jeju pony.

Results

Comparison of Metabolic Patterns in Thoroughbred and Jeju Pony Before and After Exercise

In our previous study, we conducted 1H NMR spectroscopy analysis with various equine tissue samples (plasma, muscle, and urine) following exercise [11]. In this study, we obtained plasma, urine, sweat samples from both thoroughbred and Jeju pony following exercise, as well as before exercise, and conducted 1H NMR spectroscopy analysis (Figure 1A). We obtained a very large quantity of metabolomics data. A total of 26, 39, and 36 metabolites were identified in plasma (Figure 1B), sweat (Figure 1C), and urine samples (Figure 1D), respectively. To assess which metabolites were significantly released after exercise, we compared samples obtained before and after exercise in thoroughbred and Jeju pony. Glutamate, glutamine, glutathione, lactate, and pyruvate were detected in the Jeju pony plasma samples and betaine, citrate, glucose, glutamate, glutamine, glutathione, histidine, isoleucine, leucine, phenylalanine, proline, and valine were significantly released in plasma of thoroughbred horses (Supplementary Table 1). In urine samples, trimethylamine were identified in Jeju pony and 2-oxovalerate, 3-aminoisobutyrate, alanine, citrulline, glucose, glutamine, glutarate, methylsuccinate, N-isovaleroylglycine, N-phenylacetylglycine, proline, pyruvate, taurine, threonine, tryptophan, and urea were significantly released in thoroughbred horses (Supplementary Table 2). Notably, sweat samples were difficult to collect before exercise; as such, only those collected after exercise were used (Supplementary Table 3). In addition, we analyzed metabolites that were specifically released in each tissue (Table 1). A total of 3, 12, and 15 metabolites were identified in plasma, sweat, and urine, respectively.

FIG 1

Figure 1: Venn diagram showing shared and unique metabolites (A), and heatmap analysis of the differentially expressed metabolites (B-D) in the plasma, sweat, and urine. Red and green shadings represent higher and lower relative expression levels, respectively.

Table 1: Tissue specific metabolites in both of Thoroughbred and jeju pony

Clustering

Total

Metabolites

Plasma Only 3 Glutathione, Malonate, Ornithine
Sweat Only 12 2-Hydroxybutyrate, Acetoin, Choline, Formate, Fumarate, Glycerate, Homoserine, Mannose, N-Methylhydantoin, Phenylacetate, Pyroglutamate, Urocanate
Urine Only 15 2-Oxovalerate, 3-Aminoisobutyrate, 3-Hydroxyisovalerate, Acetoacetate, Citrulline, Dimethylamine, Glutarate, Hippurate, Methylsuccinate, N-Isovaleroylglycine, N-Phenylacetylglycine, Succinate, Trimethylamine, Trimethylamine N-oxide, Tryptophan
Plasma and Sweat 22 Acetate, Alanine, Betaine, Citrate, Creatine, Glucose, Glutamate, Glycerol, Glycine, Histidine, Isoleucine, Lactate, Leucine, Lysine, Proline, Pyruvate, Serine, Threonine, Tyrosine, Valine, myo-Inositol
Sweat and Urine 20 Acetate, Alanine, Arginine, Benzoate, Creatine, Creatinine, Glucose, Glycine, Isoleucine, Lactate, Leucine, Phenylalanine, Proline, Pyruvate, Taurine, Threonine, Tyrosine, Urea, Valine, myo-Inositol
Plasma and Urine 16 Acetate, Alanine, Creatine, Glucose, Glutamine, Glycine, Isoleucine, Lactate, Leucine, Phenylalanine, Proline, Pyruvate, Threonine, Tyrosine, Valine, myo-Inositol
Plasma, Sweat, and Urine 15 Acetate, Alanine, Creatine, Glucose, Glycine, Isoleucine, Lactate, Leucine, Phenylalanine, Proline, Pyruvate, Threonine, Tyrosine, Valine, Myo-Inositol

Metabolite Set Enrichment Analyses Based on Exercise Status

Enrichment analyses of the overlapped metabolites among plasma, urine, and sweat were conducted by MetaboAnalyst 5.0 [12], and total 41 pathways were identified (Table 2). Among various pathways, the glucose-alanine cycle, glycine and serine metabolism, and alanine metabolism were the most significantly expressed after exercise.

Table 2: Enriched metabolite pathway among plasma, urine and sweat

Total

Expected

Hits

Raw p

Holm p

FDR

Glucose-Alanine Cycle

13

0.19 3 0.000667 0.0654 0.05
Glycine and Serine Metabolism

59

0.864 5 0.00103 0.1 0.05
Alanine Metabolism

17

0.249 3 0.00153 0.147 0.05
Gluconeogenesis

35

0.513 3 0.0126 1 0.308
Pyruvate Metabolism

48

0.703 3 0.0296 1 0.426
Glutamate Metabolism

49

0.718 3 0.0312 1 0.426
Glutathione Metabolism

21

0.308 2 0.0358 1 0.426
Arginine and Proline Metabolism

53

0.776 3 0.0383 1 0.426
Transfer of Acetyl Groups into Mitochondria

22

0.322 2 0.0391 1 0.426
Warburg Effect

58

0.85 3 0.0483 1 0.429
Glycolysis

25

0.366 2 0.0495 1 0.429
Valine, Leucine and Isoleucine Degradation

60

0.879 3 0.0526 1 0.429
Phenylalanine and Tyrosine Metabolism

28

0.41 2 0.0608 1 0.453
Urea Cycle

29

0.425 2 0.0647 1 0.453
Ammonia Recycling

32

0.469 2 0.0771 1 0.499
Amino Sugar Metabolism

33

0.483 2 0.0814 1 0.499
Galactose Metabolism

38

0.557 2 0.104 1 0.599
Lactose Degradation

9

0.132 1 0.125 1 0.68
Pyruvaldehyde Degradation

10

0.146 1 0.138 1 0.711
Thyroid hormone synthesis

13

0.19 1 0.176 1 0.86
Phosphatidylinositol Phosphate Metabolism

17

0.249 1 0.223 1 1
Ethanol Degradation

19

0.278 1 0.246 1 1
Catecholamine Biosynthesis

20

0.293 1 0.258 1 1
Lactose Synthesis

20

0.293 1 0.258 1 1
Threonine and 2-Oxobutanoate Degradation

20

0.293 1 0.258 1 1
Carnitine Synthesis

22

0.322 1 0.28 1 1
Cysteine Metabolism

26

0.381 1 0.322 1 1
Inositol Phosphate Metabolism

26

0.381 1 0.322 1 1
Selenoamino Acid Metabolism

28

0.41 1 0.342 1 1
Citric Acid Cycle

32

0.469 1 0.381 1 1
Inositol Metabolism

33

0.483 1 0.39 1 1
Aspartate Metabolism

35

0.513 1 0.409 1 1
Fatty Acid Biosynthesis

35

0.513 1 0.409 1 1
Porphyrin Metabolism

40

0.586 1 0.452 1 1
Sphingolipid Metabolism

40

0.586 1 0.452 1 1
Propanoate Metabolism

42

0.615 1 0.469 1 1
Methionine Metabolism

43

0.63 1 0.477 1 1
Tryptophan Metabolism

60

0.879 1 0.598 1 1
Bile Acid Biosynthesis

65

0.952 1 0.629 1 1
Tyrosine Metabolism

72

1.05 1 0.668 1 1
Purine Metabolism

74

1.08 1 0.678 1 1

Differentially Released Metabolites that Responded to Exercise in Plasma and Urine

A total of 15 metabolites, including acetate, alanine, and creatine, were observed in all sample types (plasma, urine, and sweat) (Table 1). For these metabolites, release pattern analysis after exercise was conducted in plasma and urine (Table 3). Lactate and pyruvate were significantly identified in the plasma of Jeju pony (Figure 2A) and six metabolites (glucose, isoleucine, leucine, phenylalanine, proline, and valine) were significantly identified in the thoroughbreds plasma samples (Figure 2B). In thoroughbred horses, most metabolites doubled after exercise, with glucose showing the biggest increase. Interestingly, metabolites that significantly increased after exercise in Jeju pony were showed a decreasing trend in thoroughbred horses after exercise. In addition, metabolic analysis was conducted in urine samples after exercise (Figure 3). In contrast with the plasma results, significant changes in the release of metabolites in urine were only found in the samples from thoroughbred horses. Alanine, glucose, proline, pyruvate, and threonine were significantly identified after exercise.

Table 3: Expression pattern of plasma metabolites overlapped among plasma, urine and sweat

Metabolites Jeju Horse Thoroughbreds

 

Before (Mean ±SE) mM After (Mean ±SE) mM p value Before (Mean ±SE) mM

 

After (Mean ±SE) mM

 

p value

 

Acetate 13.30 ±1.77 16.44 ±1.49 0.259 17.10±3.49 15.20±1.38 0.633
Alanine 15.26 ±1.35 17.09 ±1.82 0.49 16.39±2.46 10.78±3.18 0.248
Creatine 3.53 ±0.32 3.52 ±0.22 0.979 2.75 ± 0.41 1.79 ± 0.43 0.186
Glucose 118.24 ± 9.10 98.81 ± 5.52 0.141 104.00 ± 15.69 51.79 ± 9.07 0.0352**
Glycine 26.46 ± 2.95 23.95 ± 2.58 0.583 31.44 ± 7.79 14.75 ± 2.62 0.107
Isoleucine 2.30 ± 0.19 2.00 ± 0.24 0.4 2.46 ± 0.40 1.18 ± 0.30 0.0512*
Lactate 20.31 ± 2.14 32.04 ± 1.57 0.00418*** 18.22 ± 3.18 15.94 ± 4.46 0.719
Leucine 7.33 ± 0.22 6.62 ± 0.46 0.243 7.53 ± 1.09 3.79 ± 0.78 0.0372**
Phenylalanine 2.02 ± 0.10 2.06 ± 0.20 0.895 2.30 ± 0.30 1.23 ± 0.27 0.0436**
Proline 8.38 ± 0.73 8.76 ± 0.47 0.704 11.48 ± 1.95 5.41 ± 1.42 0.0549*
Pyruvate 0.87 ± 0.12 1.40 ± 0.15 0.0388** 0.94 ± 0.16 0.64 ± 0.11 0.21
Threonine 11.06 ± 1.94 10.60 ± 1.84 0.88 16.51 ± 2.96 9.29 ± 3.14 0.172
Tyrosine 2.75 ± 0.27 2.51 ± 0.34 0.635 3.54 ± 0.46 2.25 ± 0.57 0.133
Valine 8.68 ± 0.75 8.27 ± 1.05 0.781 11.32 ± 2.17 5.45 ± 0.79 0.0527*
Myo-Inositol 2.73 ± 0.29 2.45 ± 0.15 0.475 3.38 ± 0.53 2.35 ± 0.94 0.418

FIG 2

Figure 2: Significant difference of metabolites in plasma by exercise in Jeju pony (A) and Thoroughbreds (B). *p<0. 1, **p<0.05, ***p<0.01, ****p<0.001. All values expressed in mM as mean ± SD.

FIG 3

Figure 3: Significant difference of metabolites in urine by exercise in Thoroughbreds. *p<0. 1, **p<0.05, ***p<0.01, ****p<0.001. All values expressed in mM as mean ± SD.

Comparison of Metabolites between Equine Breeds (Thoroughbred and Jeju Pony)

In addition, we compared the metabolites between thoroughbred and Jeju pony under exercise stimuli (Tables 4-7). A greater difference was found between the metabolites released by the two breeds after exercise than before exercise in all sample types. Citrate and histidine were significantly released before exercise (Figure 4A), and 16 metabolites, including betaine and citrate, were significantly released after exercise in plasma in both breeds (Figure 4B). Among them, citrate values tripled in samples collected after exercise in both breeds and betaine and pyruvate showed largest difference between species (Figure 4B). In urine samples, six metabolites, including creatine and creatinine, showed significant differences between breeds (Figure 5). The release of taurine and myo-inositol was significantly different by more than 3.5-fold between breeds before exercise (Figure 5A) and five metabolites (creatine, creatinine, trimethylamine N-oxide, urea, and myo-inositol) were significantly different after exercise (Figure 5B). Creatine, urea, and myo-inositol more than doubled their values in urine samples after exercise (Figure 5). Although sweat samples were difficult to collect before exercise, we still analyzed sweat metabolite patterns after exercise. Among 39 metabolites, 30, including 2-hydroxybutyrate, showed significant differences between species (Table 6). Interestingly, most detected metabolites had a higher value in Jeju pony than in thoroughbred horses.

Table 4: Expression pattern of urine metabolites overlapped among plasma, urine and sweat

Metabolites

Jeju Horse

 

Thoroughbreds

 

Before (Mean ± SE) mM

 

After (Mean ± SE) mM

 

p value

 

Before (Mean ± SE) mM

 

After (Mean ± SE) mM

 

p value

 

Acetate 0.56 ± 0.14 0.41 ± 0.10 0.366 0.38 ± 0.12 0.43 ± 0.09 0.778
Alanine 0.04 ± 0.01 0.06 ± 0.02 0.573 0.03 ± 0.01 0.05 ± 0.01 0.0707*
Creatine 0.43 ± 0.32 0.09 ± 0.02 0.331 0.11 ± 0.04 0.17 ± 0.03 0.285
Glucose 0.43 ± 0.08 0.61 ± 0.10 0.285 0.27 ± 0.05 0.64 ± 0.09 0.00719***
Glycine 6.45 ± 4.42 0.39 ± 0.10 0.207 0.15 ± 0.04 0.23 ± 0.08 0.388
Isoleucine 0.06 ± 0.01 0.08 ± 0.01 0.165 0.05 ± 0.01 0.11 ± 0.03 0.107
Lactate 0.12 ± 0.04 0.13 ± 0.02 0.92 0.07 ± 0.02 0.13 ± 0.03 0.108
Leucine 0.08 ± 0.01 0.12 ± 0.02 0.308 0.10 ± 0.03 0.16 ± 0.03 0.16
Phenylalanine 0.48 ± 0.10 0.43 ± 0.08 0.793 0.31 ± 0.10 0.54 ± 0.11 0.158
Proline 0.49 ± 0.08 0.68 ± 0.17 0.2 0.32 ± 0.08 0.60 ± 0.10 0.076*
Pyruvate 0.11 ± 0.02 0.14 ± 0.05 0.71 0.08 ± 0.02 0.15 ± 0.02 0.0676*
Threonine 0.34 ± 0.10 0.19 ± 0.04 0.244 0.16 ± 0.04 0.29 ± 0.05 0.0783*
Tyrosine 0.48 ± 0.13 0.50 ± 0.11 0.924 0.36 ± 0.10 0.63 ± 0.11 0.114
Valine 0.06 ± 0.01 0.08 ± 0.01 0.349 0.07 ± 0.02 0.12 ± 0.03 0.177
Myo-Inositol 1.00 ± 0.19 1.15 ± 0.16 0.288 0.27 ± 0.06 0.41 ± 0.06 0.166

Table 5: Metabolite comparison between Thoroughbreds and jeju pony in plasma

Metabolites

Before (Mean ± SE) mM After (Mean ± SE) mM

 

TH

 

JH

 

p value

 

TH

 

JH

 

p value

 

Acetate 17.10 ± 3.49 13.30 ± 1.77 0.409 15.02 ± 1.38 16.44 ± 1.49 0.55
Alanine 16.39 ± 2.46 15.26 ± 1.35 0.729 10.78 ± 3.18 17.09 ± 1.82 0.162
Betaine 3.73 ± 1.06 3.59 ± 0.17 0.908 1.41 ± 0.34 3.17 ± 0.16 0.003***
Citrate 2.99 ± 0.36 4.09 ± 0.33 0.081** 1.48 ± 0.33 4.51 ± 0.37 0.0006****
Creatine 2.75 ± 0.41 3.53 ± 0.32 0.213 1.79 ± 0.43 3.52 ± 0.22 0.012**
Glucose 104.00 ± 15.69 118.24 ± 9.10 0.502 51.79 ± 9.70 98.81 ± 5.52 0.005***
Glutamate 9.08 ± 1.46 11.01 ± 0.67 0.315 4.07 ± 0.67 7.18 ± 0.71 0.022**
Glutamine 10.07 ± 1.89 9.98 ± 0.98 0.971 3.88 ± 0.50 7.61 ± 0.49 0.001***
Glutathione 24.70 ± 4.59 21.68 ± 1.19 0.584 12.90 ± 3.15 17.66 ± 1.52 0.259
Glycerol 3.28 ± 0.50 4.39 ± 0.27 0.118 2.64 ± 0.94 3.79 ± 0.15 0.311
Glycine 31.44 ± 7.79 26.46 ± 2.95 0.607 14.75 ± 2.62 23.95 ± 2.58 0.06*
Histidine 13.92 ± 2.01 8.56 ± 0.92 0.0621** 6.57 ± 1.55 8.70 ± 0.97 0.329
Isoleucine 2.46 ± 0.40 2.30 ± 0.19 0.759 1.18 ± 0.30 2.00 ± 0.24 0.089*
Lactate 18.22 ± 3.18 20.31 ± 2.14 0.639 15.94 ± 4.46 32.04 ± 1.57 0.016**
Leucine 7.53 ± 1.09 7.33 ± 0.22 0.877 3.79 ± 0.78 6.62 ± 0.46 0.024**
Lysine 59.40 ± 17.18 19.49 ± 9.91 0.11 39.78 ± 16.49 16.19 ± 8.54 0.289
Malonate 3.22 ± 0.42 3.98 ± 0.45 0.302 1.82 ± 0.58 3.19 ± 0.28 0.094*
Ornithine 6.46 ± 2.44 10.88 ± 3.31 0.364 3.35 ± 0.82 9.22 ± 3.22 0.152
Phenylalanine 2.30 ± 0.30 2.02 ± 0.10 0.455 1.23 ± 0.27 2.06 ± 0.20 0.056*
Proline 11.48 ± 1.95 8.38 ± 0.73 0.219 5.41 ± 1.42 8.76 ± 0.47 0.081*
Pyruvate 0.94 ± 0.16 0.87 ± 0.12 0.775 0.64 ± 0.11 1.40 ± 0.15 0.00625***
Serine 18.34 ± 3.74 15.52 ± 0.96 0.532 10.33 ± 3.30 14.46 ± 1.86 0.357
Threonine 16.51 ± 2.96 11.06 ± 1.94 0.205 9.29 ± 3.14 10.60 ± 1.84 0.756
Tyrosine 3.54 ± 0.46 2.75 ± 0.27 0.222 2.25 ± 0.51 2.51 ± 0.34 0.718
Valine 11.32 ± 2.17 8.68 ± 0.75 0.335 5.45 ± 0.79 8.27 ± 1.05 0.0917*
myo-Inositol 3.38 ± 0.53 2.73 ± 0.29 0.362 2.35 ± 0.94 2.45 ± 0.15 0.927

Table 6: Metabolite comparison between Thoroughbreds and jeju pony in Urine

Metabolites

Before (Mean ± SE) mM

 

After (Mean ± SE) mM

 

TH

 

JH

 

p value

 

TH

 

JH

 

p value

 

2-Oxovalerate 0.09 ± 0.02 0.12 ± 0.03 0.378 0.22 ± 0.05 0.14 ± 0.03 0.338
3-Aminoisobutyrate 0.28 ± 0.07 0.29 ± 0.07 0.882 0.48 ± 0.08 0.41 ± 0.12 0.851
3-Hydroxyisovalerate 0.06 ± 0.01 0.04 ± 0.00 0.34 0.10 ± 0.02 0.08 ± 0.04 0.971
Acetate 0.38 ± 0.12 0.56 ± 0.14 0.362 0.43 ± 0.09 0.41 ± 0.1 0.946
Acetoacetate 0.17 ± 0.05 0.18 ± 0.04 0.949 0.47 ± 0.20 0.21 ± 0.04 0.231
Alanine 0.03 ± 0.01 0.04 ± 0.01 0.103 0.05 ± 0.01 0.06 ± 0.02 0.628
Arginine 0.29 ± 0.10 0.43 ± 0.12 0.397 0.32 ± 0.08 0.60 ± 0.2 0.246
Benzoate 0.07 ± 0.02 5.73 ± 3.49 0.143 0.05 ± 0.01 0.06 ± 0.01 0.937
Citrulline 0.30 ± 0.07 0.41 ± 0.07 0.298 0.58 ± 0.10 0.61 ± 0.15 0.735
Creatine 0.11 ± 0.04 0.43 ± 0.32 0.342 0.17 ± 0.03 0.09 ± 0.02 0.0719*
Creatinine 11.73 ± 3.30 11.90 ± 2.37 0.968 17.00 ± 2.32 8.84 ± 2.61 0.0733*
Dimethylamine 0.13 ± 0.03 0.18 ± 0.03 0.272 0.16 ± 0.03 0.20 ± 0.03 0.34
Glucose 0.27 ± 0.05 0.43 ± 0.08 0.146 0.64 ± 0.09 0.61 ± 0.1  0.928
Glutamine 0.37 ± 0.09 0.54 ± 0.08 0.217 0.70 ± 0.13 0.53 ± 0.1 0.502
Glutarate 0.08 ± 0.02 0.12 ± 0.02 0.299 0.19 ± 0.05 0.13 ± 0.03 0.585
Glycine 0.15 ± 0.04 6.45 ± 4.42 0.191 0.23 ± 0.08 0.39 ± 0.1 0.205
Hippurate 26.02 ± 8.77 19.32 ± 3.47 0.498 53.58 ± 15.38 35.04 ± 7.85 0.381
Isoleucine 0.05 ± 0.01 0.06 ± 0.01 0.814 0.11 ± 0.03 0.08 ± 0.01 0.313
Lactate 0.07 ± 0.02 0.12 ± 0.04 0.266 0.13 ± 0.03 0.13 ± 0.02 0.976
Leucine 0.10 ± 0.03 0.08 ± 0.01 0.553 0.16 ± 0.03 0.12 ± 0.02 0.405
Methylsuccinate 0.14 ± 0.04 0.15 ± 0.02 0.814 0.30 ± 0.07 0.21 ± 0.05 0.474
N-Isovaleroylglycine 0.10 ± 0.02 0.13 ± 0.03 0.502 0.16 ± 0.01 0.14 ± 0.03 0.831
N-Phenylacetylglycine 5.94 ± 1.38 7.40 ± 1.50 0.494 10.96 ± 1.83 8.63 ± 1.34 0.524
Phenylalanine 0.31 ± 0.10 0.48 ± 0.10 0.264 0.54 ± 0.11 0.43 ± 0.08 0.687
Proline 0.32 ± 0.08 0.49 ± 0.08 0.198 0.60 ± 0.10 0.68 ± 0.17 0.466
Pyruvate 0.08 ± 0.02 0.11 ± 0.02 0.44 0.15 ± 0.02 0.14 ± 0.05 0.998
Succinate 0.03 ± 0.01 0.04 ± 0.01 0.197 0.05 ± 0.01 0.03 ± 0.01 0.419
Taurine 0.23 ± 0.06 0.86 ± 0.23 0.0312** 0.78 ± 0.17 1.16 ± 0.41 0.738
Threonine 0.16 ± 0.04 0.34 ± 0.10 0.127 0.29 ± 0.05 0.19 ± 0.04 0.3
Trimethylamine 0.03 ± 0.01 0.04 ± 0.01 0.138 0.04 ± 0.00 0.02 ± 0 0.198
Trimethylamine N-oxide 0.15 ± 0.05 0.20 ± 0.06 0.553 0.11 ± 0.02 0.22 ± 0.03 0.025**
Tryptophan 0.26 ± 0.07 0.29 ± 0.03 0.69 0.50 ± 0.09 0.47 ± 0.08 0.973
Tyrosine 0.36 ± 0.10 0.48 ± 0.13 0.512 0.63 ± 0.11 0.50 ± 0.11 0.636
Urea 84.58 ± 16.17 79.19 ± 9.54 0.782 173.25 ± 11.44 103.46 ± 9.89 0.00177***
Valine 0.07 ± 0.02 0.06 ± 0.01 0.842 0.12 ± 0.03 0.08 ± 0.01 0.322
Myo-Inositol 0.27 ± 0.06 1.00 ± 0.19 0.0062*** 0.41 ± 0.06 1.15 ± 0.16 0.00639***

Table 7: Metabolite comparison between Thoroughbreds and jeju pony in Sweat

Metabolites

Before (Mean ± SE) mM

 

TH

 

JH

 

p value

 

2-Hydroxybutyrate 0.05 ± 0.01 0.28 ± 0.08 0.0505*
Acetate 1.06 ± 0.42 2.15 ± 0.29 0.331
Acetoin 0.00 ± 0.00 0.05 ± 0.00 1.07e-05****
Alanine 0.14 ± 0.05 1.40 ± 0.37 0.0281**
Arginine 0.06 ± 0.02 1.06 ± 0.48 0.108
Benzoate 0.03 ± 0.01 0.26 ± 0.10 0.095*
Betaine 0.01 ± 0.00 0.06 ± 0.02 0.0602*
Choline 0.00 ± 0.00 0.01 ± 0.00 0.00326***
Citrate 0.49 ± 0.06 8.16 ± 3.08 0.0693*
Creatine 0.03 ± 0.01 0.18 ± 0.01 0.000221****
Creatinine 0.07 ± 0.03 0.12 ± 0.04 0.532
Formate 0.18 ± 0.04 0.49 ± 0.02 0.00646***
Fumarate 0.01 ± 0.00 0.05 ± 0.01 0.0561*
Glucose 0.17 ± 0.06 1.55 ± 0.36 0.0223**
Glutamate 0.08 ± 0.01 0.45 ± 0.08 0.00996***
Glycerate 0.07 ± 0.02 0.31 ± 0.02 0.00222***
Glycerol 0.10 ± 0.02 1.18 ± 0.20 0.00578***
Glycine 0.14 ± 0.05 1.70 ± 0.47 0.0299**
Histidine 0.02 ± 0.00 0.63 ± 0.41 0.215
Homoserine 0.06 ± 0.01 0.33 ± 0.12 0.114
Isoleucine 0.03 ± 0.01 0.20 ± 0.06 0.0457**
Lactate 0.43 ± 0.14 5.90 ± 1.92 0.0519*
Leucine 0.04 ± 0.01 0.23 ± 0.06 0.0324**
Lysine 0.02 ± 0.01 0.19 ± 0.06 0.057*
Mannose 0.10 ± 0.03 0.16 ± 0.01 0.0367**
N-Methylhydantoin 0.00 ± 0.00 0.03 ± 0.01 0.0339**
Phenylacetate 0.02 ± 0.00 0.07 ± 0.03 0.129
Phenylalanine 0.02 ± 0.00 0.21 ± 0.06 0.0362**
Proline 0.06 ± 0.01 0.19 ± 0.04 0.0648*
Pyroglutamate 0.15 ± 0.05 1.61 ± 0.58 0.0639*
Pyruvate 0.10 ± 0.03 0.84 ± 0.01 1.44e-05****
Serine 0.18 ± 0.06 2.77 ± 1.41 0.138
Taurine 0.02 ± 0.00 0.08 ± 0.01 0.000513****
Threonine 0.04 ± 0.01 0.53 ± 0.27 0.15
Tyrosine 0.02 ± 0.00 0.13 ± 0.04 0.0503*
Urea 10.05 ± 1.98 7.93 ± 3.51 0.894
Urocanate 0.04 ± 0.01 0.24 ± 0.06 0.0283**
Valine 0.04 ± 0.01 0.26 ± 0.08 0.0506*
Myo-Inositol 0.06 ± 0.01 0.25 ± 0.04 0.0134**

FIG 4

Figure 4: Significant difference of metabolites in plasma between breeds (Jeju pony and Thoroughbreds) before (A) and After exercise (B). *p<0. 1, **p<0.05, ***p<0.01, ****p<0.001. All values expressed in mM as mean ± SD.

FIG 5

Figure 5: Significant difference of metabolites in urine between breeds (Jeju pony and Thoroughbreds) before (A) and after exercise (B). *p<0. 1, **p<0.05, ***p<0.01, ****p<0.001. All values expressed in mM as mean ± SD.

Discussion

Almost 60 million horses currently exist on the planet. In addition to providing important services such as transport, meat, leather, and ploughing force and in the majority of developing countries, horses are mainly used for sports and leisure activities in most developed countries [13]. Therefore, as one of their most important economic traits, most research conducted in horses focuses on improving their athletic abilities [14,15]. However, although their physical and physiological adaptations receive much attention [16], targeted genes and metabolites or underlying mechanisms associated with exercise are still understudied.

The advances in metabolic analysis technology that have been carried out allow the assessment of the physiological state of individuals [17] and prediction of their condition [18]. Therefore, metabolomics demonstrates various biological responses to environmental influences, genetic, transcriptomic, and proteomic, [19-21]. Because of these advantages, metabolic analysis is widely used to explore metabolic patterns [22] or to discover new biomarkers through physical changes associated with diseases or environmental changes [21,23].

Although previous studies have investigated the metabolic changes caused by exercise, most only analyzed skeletal muscle [24] and were further limited by their small sample size and little expansive metabolite platform [25]. Previous metabolic studies on exercise mainly focus on the effect of exercise in various tissues [26,27], and studies on the discovery of biomarkers, which are affected by the athletic ability of individuals, are relatively poorly performed. Jang et al., 2017, the basis of this study, conducted a metabolic analysis in skeletal muscle, plasma, and urine samples after exercise [11]. In this study, we performed a metabolic analysis in plasma, urine, and sweat samples of thoroughbred and Jeju pony by exercise. In addition, we demonstrated the influence of exercise and breed in metabolite levels. We obtained a large amount of metabolite data that were released after exercise. Among 15 metabolites that were commonly detected in plasma, urine, and sweat, the levels of lactate, pyruvate, glucose, isoleucine, leucine, phenylalanine, proline, and valine showed significantly changes after exercise in plasma samples (Figure 2), and the levels of alanine, glucose, proline, pyruvate, and threonine had significantly changed after exercise in urine samples (Figure 3). These results are in line with those of previous studies [11]. The metabolites observed in samples collected after exercise were all associated with the tricarboxylic acid (TCA) cycle, with some being intermediate products. Alanine, aspartate, and glutamate metabolism and aminoacyl-tRNA and arginine biosynthesis related metabolic pathways are activated by acute exercise [28]. These results suggest that several metabolic pathways that utilize skeletal muscle substrate are regulated after exercise, and previous studies reported that this occurs in various tissues [29,30].

During exercise, muscle glycogen, its main source of energy, is altered to glucose and subsequently to pyruvate via glycolysis [31]. The pyruvate converted by glycolysis can enter TCA and glucose-alanine cycles or be converted to lactate [32]. During aerobic exercise, muscle glycogen can be used to produce ATP through glycolysis; however, when anaerobic exercise like a sprint is conducted, the muscles cannot use oxygen for glycolysis [33]. Therefore, muscle glycogen (glucose) is altered to lactate through anaerobic glycolysis [33]. Then, the lactate is released to the bloodstream and transferred to the kidneys and liver [34]. In the liver, lactate is altered to pyruvate through gluconeogenesis [35]. In addition, when amino acids are used for energy in extrahepatic tissues, pyruvate derived from the glycolysis is used as an amino group receptor to form alanine, a non-essential amino acid [36]. The produced alanine is transferred to the liver through the bloodstream and converted to either pyruvate for gluconeogenesis via the glycose–alanine cycle or to glutamate, which then goes through the urea cycle. Collectively, the detected metabolites in equine plasma and urine including glucose, alanine, and lactate were altered to pyruvate and used for energy production. Therefore, the metabolites discovered in this study can be used as a reasonable indicator to measure athletic ability and exercise fatigue.

In conclusion, we compared metabolite presence between thoroughbreds and Jeju pony after exercise and analyzed enriched metabolic pathways of commonly detected metabolites in all samples (plasma, urine, and sweat). Our results could help improve our understanding of exercise fatigue and find regulation markers for fatigue reduction. Further research is necessary to combine these results with other omics data and reveal the function of metabolic markers.

Declarations

Ethics Approval and Consent to Participate

All animal procedures used in the study were conducted in compliance with international standards and were approved by the Institutional Animal Care and Use Committee of Pusan National University (Approval Number: PNU-2015-0864).

Competing Interests

The authors declare that they have no competing interests.

Acknowledgments

This work was supported by a 2-Year Research Grant from the Pusan National University.

Author’s Contribution

The research was conceptualized by Park JW, Cho BW and further edition was done by all the authors.  Data was curated by Park JW, Kim KH, and analyzed by Park JW, Lee SI, Sang SS. All authors have participated on data interpretation. The draft of the manuscript was written by Park JW and Kim KH, and the final form was edited by Lee SI, Sang SS, and Cho BW. All authors have contributed by interpretation, analysis, critical discussion.

References

  1. Kim H, Lee T, Park W, Lee JW, Kim J, et al. (2013) Peeling back the evolutionary layers of molecular mechanisms responsive to exercise-stress in the skeletal muscle of the racing horse. DNA Res 20: 287-298. [crossref]
  2. PARK Jeong-Woong, Choi JY, Hong SA, Kim NY, Do KT, et al. (2017) Exercise induced upregulation of glutamate-cysteine ligase catalytic subunit and glutamate-cysteine ligase modifier subunit gene expression in Thoroughbred horses. Asian-Australasian Journal of animal sciences 30: 728-735. [crossref]
  3. CHO Hyun-Woo, Shin S, Park JW, Choi JY, Kim NY, et al. (2015) Molecular characterization and expression analysis of the peroxisome proliferator activated receptor delta (PPARδ) gene before and after exercise in horse. Asian-Australasian Journal of Animal Sciences 28: 697. [crossref]
  4. KHUMMUANG Saichit, Lee HG, Joo SS, Park JW, Choi JY, et al. (2020) Comparison for immunophysiological responses of Jeju and Thoroughbred horses after exercise. Asian-Australasian Journal of Animal Sciences 33: 424-435. [crossref]
  5. PARK Jeong-Woong, Kim KH, Choi JK, Park TS, Song KD, et al. (2021) Regulation of toll-like receptors expression in muscle cells by exercise-induced stress. Animal Bioscience 34: 1590. [crossref]
  6. LEE Hyo Gun, Choi JY, Park JW, Park TS, Song KD, et al. (2019) Effects of exercise on myokine gene expression in horse skeletal muscles. Asian-Australasian Journal of Animal Sciences 32: 350-356. [crossref]
  7. Jordan Kate W, Nordenstam J, Lauwers GY, Rothenberger DA, Alavi K, et al. (2009) “Metabolomic characterization of human rectal adenocarcinoma with intact tissue magnetic resonance spectroscopy.” Diseases of the colon and rectum 52: 520. [crossref]
  8. Hargreaves Mark (2000) “Skeletal muscle metabolism during exercise in humans.” Clinical and Experimental Pharmacology and Physiology 27: 225-228. [crossref]
  9. Moghetti Paolo, Bacchi E, Brangani C, Donà S, Negri C (2016) “Metabolic effects of exercise.” Sports Endocrinology 47: 44-57. [crossref]
  10. Jang Hyun-Jun, Kim DM, Kim KB, Park JW, Choi JY, et al. (2017) “Analysis of metabolomic patterns in thoroughbreds before and after exercise.” Asian-Australasian journal of animal sciences 30: 1633-1642. [crossref]
  11. PANG Zhiqiang, Zhou G, Ewald J, Chang L, Hacariz O, et al. (2022) Using MetaboAnalyst 5.0 for LC–HRMS spectra processing, multi-omics integration and covariate adjustment of global metabolomics data. Nature Protocols 17: 1735-1761. [crossref]
  12. ORLANDO Ludovic (2020) The evolutionary and historical foundation of the modern horse: Lessons from ancient genomics. Annual Review of Genetics 54: 563-581. [crossref]
  13. Wilkin Tessa, Anna Baoutina, Natasha Hamilton (2017) “Equine performance genes and the future of doping in horseracing.” Drug testing and analysis 9: 1456-1471. [crossref]
  14. Arfuso Francesca, Assenza A, Fazio F, Rizzo M, Giannetto C, et al. (2019) “Dynamic change of serum levels of some branched-chain amino acids and tryptophan in athletic horses after different physical exercises.” Journal of equine veterinary science 77: 12-16. [crossref]
  15. Arfuso Francesca, Giudice E, Panzera M, Rizzo M, Fazio F, et al. (2022) “Interleukin-1Ra (Il-1Ra) and serum cortisol level relationship in horse as dynamic adaptive response during physical exercise.” Veterinary Immunology and Immunopathology 243: 110368. [crossref]
  16. SABATINE Marc S, Liu E, Morrow DA, Heller E, McCarroll R, et al. (2005) Metabolomic identification of novel biomarkers of myocardial ischemia. Circulation 112: 3868-3875. [crossref]
  17. SHAH Svati H, Bain JR, Muehlbauer MJ, Stevens RD, Crosslin DR, et al. (2010) Association of a peripheral blood metabolic profile with coronary artery disease and risk of subsequent cardiovascular events. Circulation: Cardiovascular Genetics 3: 207-214. [crossref]
  18. GERMAN J Bruce, HAMMOCK Bruce D, WATKINS Steven M (2005) Metabolomics: building on a century of biochemistry to guide human health. Metabolomics 1: 3-9. [crossref]
  19. OREŠIČ Matej, VIDAL-PUIG Antonio, HÄNNINEN Virve (2006) Metabolomic approaches to phenotype characterization and applications to complex diseases. Expert review of molecular diagnostics 6: 575-585. [crossref]
  20. MOORE Rowan E, Kirwan J, Doherty MK, Whitfield PD (2007) Biomarker discovery in animal health and disease: the application of post-genomic technologies. Biomarker insights 2: 185-196. [crossref]
  21. KELL Douglas B (2004) Metabolomics and systems biology: making sense of the soup. Current opinion in microbiology 7: 296-307. [crossref]
  22. WHITFIELD, Phillip David, et al. (2005) Metabolomics as a diagnostic tool for hepatology: validation in a naturally occurring canine model. Metabolomics 1: 215-225.
  23. HUFFMAN Kim M, Koves TR, Hubal MJ, Abouassi H, Beri N, et al. (2014) Metabolite signatures of exercise training in human skeletal muscle relate to mitochondrial remodelling and cardiometabolic fitness. Diabetologia 57: 2282-2295. [crossref]
  24. HUFFMAN Kim M, Slentz CA, Bateman LA, Thompson D, Muehlbauer MJ, et al. (2011) Exercise-induced changes in metabolic intermediates, hormones, and inflammatory markers associated with improvements in insulin sensitivity. Diabetes Care 34: 174-176. [crossref]
  25. BRENNAN Andrea M, Benson M, Morningstar J, Herzig M, Robbins J, et al. (2018) Plasma metabolite profiles in response to chronic exercise. Medicine and science in sports and exercise 50: 1480-1486. [crossref]
  26. BRENNAN Andrea M, Tchernof A, Gerszten RE, Cowan TE, Ross R (2018) Depot-specific adipose tissue metabolite profiles and corresponding changes following aerobic exercise. Frontiers in Endocrinology 9: 759. [crossref]
  27. TABONE Mariangela, et al. (2021) The effect of acute moderate-intensity exercise on the serum and fecal metabolomes and the gut microbiota of cross-country endurance athletes. Scientific reports 11: 1-12.
  28. MANAF Faizal A, Lawler NG, Peiffer JJ, Maker GL, Boyce MC, et al. (2018) Characterizing the plasma metabolome during and following a maximal exercise cycling test. Journal of Applied Physiology 125: 1193-1203. [crossref]
  29. HEINONEN Ilkka, Kalliokoski KK, Hannukainen JC, Duncker DJ, Nuutila P, et al. (2014) Organ-specific physiological responses to acute physical exercise and long-term training in humans. Physiology 29: 421-436. [crossref]
  30. SPRIET Lawrence L (2002) Regulation of skeletal muscle fat oxidation during exercise in humans. Medicine and science in sports and exercise9: 1477-1484. [crossref]
  31. ISHIKURA, Keisuke; RA, Song-Gyu, OHMORI, Hajime (2013) Exercise-induced changes in amino acid levels in skeletal muscle and plasma. The Journal of Physical Fitness and Sports Medicine3: 301-310.
  32. HIGGINS, Chris. Lactate and lactic acidosis. 2007. [crossref]
  33. YANG Woo-Hwi, Park H, Grau M, Heine O (2020) Decreased blood glucose and lactate: is a useful indicator of recovery ability in athletes? International Journal of Environmental Research and Public Health15: 5470. [crossref]
  34. CHIANG J (2014) Liver physiology: Metabolism and detoxification.
  35. EXTON JH, Mallette LE, Jefferson LS, Wong EH, Friedmann N, et al. (1970) The hormonal control of hepatic gluconeogenesis. In: Proceedings of the 1969 Laurentian Hormone Conference. Academic Press 411-461. [crossref]
  36. JOSE Caroline, Melser S, Benard G, Rossignol R (2013) Mitoplasticity: adaptation biology of the mitochondrion to the cellular redox state in physiology and carcinogenesis. Antioxidants & Redox Signaling7: 808-849. [crossref]

Errors during the Night Shift: An Age Management Policy is Needed for Nurses Over 55

DOI: 10.31038/IJNM.2023431

 
 

In the study “Relation between sleep deprivation and nursing errors during the night shift,” conducted on a sample of 3358 nurses, 94.7% of the respondents stated that they perform regular activities during the night shift. 16.8% of the respondents made errors in the last two night shifts, with 59.5% of the cases involving a single error. Sleep deprivation (3-5 hours of sleep within the 24 hours preceding the interview) appears to contribute to a higher frequency of errors. This tendency is particularly pronounced among nurses working in critical care units (23.9%). The number of nights worked also influences the likelihood of making errors, with an error frequency of 20.5%. The propensity to make errors is relatively high among the younger age group (17.2%), decreases in the middle age group (15.8%), and then increases again in the older age group (17.6%). We have concluded that night-time care is exhausting and demanding and can have negative effects on the quality of care provided. Therefore, healthcare organizations should support nurses in the organization of night-time care by investing in strategies for safe and quality night-time care that minimizes the negative impact on nurses’ quality of life and psycho-physical well-being. This includes increasing the number of nursing staff during the night shift. Furthermore, the authors conclude that precise age management strategies supported at the management level are needed to minimize the negative effects of the current situation and to enhance the skills that increase in quantity and quality with work experience, utilizing them as valuable resources within the system.

A significant portion of the global and Italian nursing workforce is represented by those over 55. According to the “Health Profile 2019” for Italy published by the OECD and the European Commission in 2020, available on the FNOPI website, the majority of nurses are concentrated in the age group between 36 and 55 years: 268,914. There are also 15,552 nurses between 20 and 25 years old and 13,259 nurses over 65. The “younger” nurses (up to 58 years old with over 30 years of professional experience) number over 30,000. Nurses over 60 (with over 30 years of professional experience) amount to just over 13,000, and nurses up to 28 years old, at risk of underemployment/unemployment, are 39,000. Finally, nurses over 60 without more than 30 years of professional experience amount to approximately 25,000. Nurses over 55 are an important segment of the nursing profession who, along with advancing age and years of service, face a range of psychophysical issues that make it difficult to ensure efficient work performance. Despite this, healthcare companies do not pay attention to this age group of nurses. What would nurses want for their careers in the years leading up to retirement?

A survey conducted at the Modena Local Health Authority investigated the organizational health status of workers over 50, their perceptions, motivations, health issues, and proposed measures to raise awareness of the phenomenon of aging in the profession and suggest improvement actions to be tested within the company. Nurses who participated in the survey listed some self-strategies to mitigate the impact of aging and strategies to suggest to the organizational system.

Self-strategies:

  • Invest in personal professional development and participate in individual training courses.
  • Maintain a part-time employment contract.
  • Have more hours of rest.
  • Achieve a balanced use of absences from work for psycho-physical recovery by reviewing the threshold for illness and the use of contractually defined leaves.
  • Request authorization for extended ordinary leave of at least 4-5 consecutive days for rest and better vacation planning.
  • Strategies to suggest to the organizational system:
  • Value the experience gained by older nurses by combining the energy and vitality of younger nurses with the knowledge and experience of older ones. The value of older nurses should be measured by recognizing their wealth of skills and field experience, different culture, and being the key to the company’s historical memory, as older nurses are loyal to their work due to cultural and generational factors.
  • Identify older nurses who can facilitate the integration of newly hired nurses.
  • Mentoring, the process of knowledge transfer through the support of newly hired or nursing students, can play a significant role in work organization.
  • Motivate older nurses by bridging their technological training gaps. Older nurses today face greater difficulty in adapting to new technologies.
  • Employ older nurses in less stressful services with lower workloads, services that require experience and good relational skills more than physical endurance.
  • Exempt employees over 55, upon request, from night shifts and on-call shifts, and ensure a regular schedule to allow for psycho-physical recovery.

Grandparents’ Family Functions in Grandchildrearing in Japan, and Its Effect Factors

DOI: 10.31038/IJNM.2023424

Abstract

Midwives have the role of helping grandparents enhance their family functions as they welcome new family members. However, their specific functions and effect factors remain unclear. Therefore, we conducted a survey with the aim of clarifying the family function of grandparents in raising grandchildren. We conducted an online survey of 2,000 Japanese grandparents and analyzed the data using factor analysis and multiple regression.

Four functions of the grandparents were revealed: daily care and healthcare, emotional support for parents, grandchildren’s character development, and own well-being. Raising grandchildren exerts a great influence on family dynamics. Grandparents who did not feel burdened reported that grandchild-rearing was one of their lives’ passions and helped them maintain their strength and youth. To enhance their family functions grandparents, it is necessary to assess the four functions of grandparents, support them in maintaining an appropriate distance from their grandchildren and parents, and set up a forum for discussion between grandparents and parents.

Keywords

Childcare, Family function, Grandchildren, Grandparents

Contribution to the Field

  • The family function of grandparents in raising Japanese grandchildren was clarified. Therefore, in the future, the viewpoints of the four family functions mentioned above will be used as an assessment tool, which may lead to the implementation of midwives’ care that enhances the family functions of grandparents.
  • To improve the quality of life of grandparents, the need to maintain a moderate distance from grandchildren was revealed. New grandparents of both sexes can be educated in advance about the role of grandparents.

Introduction

Against the backdrop of declining family functions, problems such as depression, suicide among pregnant women [1], and infant abuse continue to persist [2]. The birth of a new child represents a developmental crisis because the family roles of both parents and grandparents change considerably. The main purpose of family support is to enable families welcoming new children to smoothly fulfill their roles and functions and enhance family functions. In Japan, for grandparents, the birth of a grandchild is the achievement of family succession, and the involvement of grandparents in childcare improves the grandparents’ own quality of life [3,4]. For mothers, the influence of grandparents has the merits of physical recovery after childbirth, elimination of anxiety about childcare, and passing on childcare skills [5]. In recent years, the employment rate of Japanese mothers has increased [6]. Mothers need access to formal childcare schemes [7] and informal family support to balance childcare and work. The Japanese tend to choose informal support from grandparents when they need childcare support [8,9]. Furthermore, 53% of Japanese couples receive childcare support from their mothers until their children are three years old. If wives are employed, this number rises to 58% [6]. In addition, the role of grandparents as caretakers of grandchildren is associated with an increase in the number of births [10]. All of this indicates a growing expectation that grandparents will take on a role in providing childcare for their grandchildren. However, involvement in grandchild-rearing adds a physical and emotional burden on grandparents [11,12]. Differences in childcare policies between mothers and grandparents have created issues such as burden and stress on mothers [5,13]. In the future, as the average life expectancy increases further [14] it is conceivable that as grandparents get older, the physical burden will increase, making it more difficult for them to be involved in the regular rearing of their grandchildren [15]. Midwives have a role to play in helping families, including grandparents, adapt to the birth of a new child [16]. Therefore, it is necessary to assess the family functions of grandparents and provide appropriate support. In recent years, grandparents who welcome new grandchildren have been educated about childcare and the roles of grandparents so that they can understand the role and take charge of raising their grandchildren [17]. However, these supports are not sufficient to meet the needs of grandparents because they are care plans based on an assessment of the family functions of grandparents raising grandchildren and remain ambiguous. Previous studies have cited the role of grandparents as providing basic care to their grandchildren, ensuring their safety and health [18] and identifying themselves as role models and educators [18-20]. In addition, the extent and impact of grandparents’ involvement in parenting have been clarified, but the exact family functions of grandparents remain unknown. Grandparents often refer to the ambiguity of their role within the family and the concomitant challenge of adjusting the parents’ roles, which can lead to the drawing of boundaries and severing of family ties [21]. Therefore, it is necessary to clarify the role function of grandparents responsible for raising grandchildren to support the family so that they can perform individual functions in the family without crossing boundaries. Accordingly, this study aimed to determine grandparents’ family functions in grandchild-rearing in Japan and factors that influence them.

Methods and Materials

Operational Definitions and Conceptual Framework

Grandparents

Men and women with grandchildren between 0 and 5 years of age, with the infants’ parents being over the age of 40.

Grandparents’ Family Functions

Grandparents’ family function was defined as the state in which grandparents maintain and improve their own well-being in harmony with their work, hobbies, and community activities while building relationships with their own children and their children’s families [22].

Research Design

This cross-sectional observational study used a questionnaire survey.

Participants

The sample comprised 2,000 individuals (50% women), 40–89 years of age, with preschool-aged grandchildren with whom they interacted at least once a year.

Survey Period and Methodology

In July 2021, we commissioned a Japanese internet research firm to conduct an online survey, ensuring participants’ anonymity. The sample was selected from a list of people registered with this firm. The internet research company’s monitors are registered in accordance with the rules and have 1.12 million monitors. Participants were selected from among them.

Survey Contents

Participants’ Attributes and the Burden of Raising a Grandchild

The participant attributes surveyed included gender, age, cohabitation, and employment. The respondents who indicated being “very” (scored as 4) or “somewhat” involved (scored as 3) in their grandchildren’s development, supporting the parents, and general housework, were defined as “involved.” Those who indicated being “not very involved” (scored as 2) and having “almost no involvement” (scored as 1) were defined as “not involved.” Regarding the burden of child-rearing (e.g., caring for their grandchildren, disciplining them, supporting their parents, and doing general housework), those who indicated feeling “very” (scored as 4) and “somewhat” (scored as 3) burdened, were assigned to the “burden” group, whereas those who indicated feeling “not very burdened” (scored as 2) and “almost no burden” (scored as 1) were assigned to the “no burden” group.

Grandparents’ Family Functions

In order to clarify the family function of grandparents, 15 items that assessed grandparents’ family functions were extracted from previous studies [11,23-25]. Experts examined the constructs of the extracted items. Items were scored on a five-point Likert scale, ranging from 1 (“not at all applicable”) to 5 (“very applicable”).

Analysis Method

Basic statistics were calculated for each variable, and an exploratory factor analysis was conducted for the grandparents’ family function items. After confirming multicollinearity using the correlation coefficient variance inflation factor (VIF), a linear regression analysis (i.e., forced entry method) was conducted with the grandparents’ family functions as the dependent variables and basic attributes as the independent variables. Multiple regression analysis was conducted between the grandparents’ family functions subscale and basic attributes. Gender was coded as 0=male and 1=female. Employment and cohabitation status were both coded as 0=no and 1=yes. The dummy variables for involvement in grandchild-rearing and the degree of grandchild burden were set as 0=none and 1=yes. The significance level was set at 5%. Statistical analyses were performed using IBM’s SPSS Statistics software, Ver. 27.

Ethical Considerations

A statement clearly stating the ethical considerations (e.g., guarantee of voluntary research cooperation and anonymity, confidential handling of data after completion of the research, existence of conflicts of interest, and monitoring) was displayed on the screen before the beginning of the survey. The study was approved by the Ethical Review Committee for Nursing Research of Dokkyo Medical University (approval number: Nursing 03008). Because this was an online survey, the study participants provided informed consent by checking a box to indicate that they agree with the ethical statements that appeared on the screen.

Results

Overview of the Participants

A summary of the participants’ characteristics is presented in Table 1. The participants’ age averaged 63.4 years (6.9 SD); 67 of them (3.4%) were in their 40s, 451 (22.6%) were in their 50s, 1,092 (54.6%) were in their 60s, and 390 (19.5%) were in their 70s or older. Further, 715 participants (35.8%) lived with their grandchildren, whereas 1,285 (64.3%) did not. Additionally, 1,088 (54.4%) were involved in grandchild care, 1,085 (54.3%) were involved in supporting their own children, and 1,102 (55.1%) were not involved in general housework. Of the respondents, 1,248 (62.3%) did not feel burdened by grandchild care, 1,211 (60.6%) did not feel burdened by disciplining grandchildren, 1,624 (81.2%) did not feel burdened by supporting their own children, and 1,102 (55.1%) did not feel burdened by general housework.

Table 1: Outline of eligible persons (N=2,000)

TAB 1

Notes: 1Involvement in child-rearing: Yes (very involved, somewhat involved); No (not very involved, almost no involvement).
2Burden of child-rearing: Yes (very burdened, somewhat burdened); No (not very burdened, no burden).

Grandparents’ Family Functions and Effect Factors

The 15 items of grandparents’ family functions were confirmed by calculating basic statistics; no ceiling or floor effects were found. These items were subsequently subjected to exploratory factor analysis, based on the maximum likelihood method. Judging from the possibility of interpretation, a factor analysis of Promax rotation was performed with four factors. As a result, two items, “I am troubled by the gap between my own child-rearing experience and that of my own children” and “I try to ease the strained relationship between my grandchildren and their parents,” were deleted because their factor loadings were lower than 0.4, and factor analysis was conducted again using maximum likelihood method and Promax rotation. Table 2 presents the factor loadings.

Table 2: Grandparents’ family function: exploratory factor analysis (N=2,000)

TAB 2

Notes: Maximum likelihood method: Promax method with Kaiser normalization.
Kaiser-Meyer-Olkin .93; Bartlett spherical test of: p<0.001.

The following four dimensions of grandparents’ family function were identified: Function 1 was named daily care and health care because of its high loadings in “contributing to grandchildren’s health and growth by helping them eat and bathe,” “contributing to grandchildren’s health recovery by taking care of them when they are sick,” “taking time away from work and hobbies to take care of them,” “contributing to the stability of family life for the parental couple,” and “providing relaxation time for the parental couple by taking care of the grandchildren”. Function 2 was named emotional support for parents due to the high loadings of “I respect the parenting policies of my own children and am involved in my grandchildren’s lives,” “I watch the human development of my own children through child-rearing,” and “I ask myself whether the degree to which I help my children and their spouses with child-rearing is excessive”. Function 3 was named grandchildren’s character development owing to its high loadings for “I help my grandchildren develop lifestyle habits and learn social rules”. “I help my grandchildren learn compassion and patience,” and “I am a good role model for my grandchildren”. Function 4 was named own well-being because of its high loadings for the items “raising grandchildren is one of the things that makes life worth living,” and “being involved in my grandchildren’s lives helps me maintain my own physical strength and youthfulness”. Cronbach’s α coefficients for dimensions 1–4 were 0.91, 0.86, 0.78, 0.81, and 0.86, respectively. The inter-factor correlations are depicted in Table 3.

Table 3: Grandparents’ family function: correlation between factors

TAB 3

Note: Maximum likelihood method: Promax method with Kaiser normalization.

Linear regression analysis of grandparents’ family functions and participants’ characteristics is presented in Table 4. Multiple regression analysis was performed, with grandparents’ family functions as dependent variables and basic attributes as independent variables. All variables were included because there were no variables with r>0.8. The VIFs were all lower than 10.0; therefore, there were no problems with multicollinearity.

Table 4: Comparison of grandparents’ family function and grandparents’ attributes

TAB 4

Notes: SCs: Standardized Coefficients; NSCs: Non-Standardized Coefficients.

Function 1 was significantly affected by grandchildren’s involvement in childcare: (β=0.392, p<0.000), support for the parents (β=0.189, p<0.000), general housework (β=0.181, p<0.000), co-residence (β=0.058, p<0.003), and age (β=-0.048, p<0.009) had a significant effect.

Function 2 was significantly affected by support for their own children: (β=0.203, p<0.000), involvement in raising the grandchildren (β=0.102, p<0.000), age (β=0.056, p<0.017), and the burden of support for their own children (β=-0.066, p<0.009), and gender (β=-0.096, p<0.000).

Function 3 included involvement in childcare (β=0.285, p<0.000), support for the parents (β=0.215, p<0.000), general housework (β=0.144, p<0.000), cohabitation (β=0.074, p<0.001), gender (β=0.067, p<0.002), the burden of general housework (β=0.053, p<0.018), and the burden of support for parents (β=0.048, p<0.028). The burden of general housework (β=0.053, p<0.018), burden of support for parents (β=0.048, p<0.028), and burden of disciplining grandchildren (β=-0.084, p<0.000) had significant effects.

Function 4 was significantly affected by involvement in childcare: (β=0.284, p<0.000), support for parents (β=0.151, p<0.000), general housework (β=0.099, p<0.000), gender (β=0.080, p<0.001), and the burden of childcare (β=-0.146, p<0.000).

Discussion

Characteristics of the Target Population

Over 50% of participants were involved in grandchild-rearing, which is in line with the finding that 53% of Japanese couples receive childcare support from their grandmothers by the time their children are three years old [6] half of grandparents in China [26], and 42-44% of grandparents in Europe and other countries take care of their grandchildren [27]. In the United States, 25% of children under five years of age are in the care. Thus, it can be said that the subjects of this study are a general population.

Furthermore, 64.3% of participants did not live with their grandchildren. In a survey of Japanese people’s perceptions regarding cohabitation, 22% of respondents stated that living apart from their grandparents was preferred, but that living in the same neighborhood as their parents was ideal [28]. This suggests that Japanese people prefer to maintain a moderate distance from their parents. The reason for this may be that, for the parental generation, grandparents provide childcare support when needed, which is beneficial to both the parental and grandparental generations’ psychological health.

Grandparents’ Family Functions

The four family functions of grandparents were extracted: daily care and health care, emotional support for parents, grandchildren’s character development, and own well-being. Shiraishi and Inoue (2017) categorized grandparents’ participation in grandchild care as support for grandchildren’s daily life, support for parents’ daily life, emotional support for parents, and support for grandchildren’s emotional and social needs [29]. The three functions revealed in this study: daily care and health care, emotional support for parents, and grandchildren’s character development were consistent with Shiraishi and Inoue (2017) [29]. According to a survey by the Cabinet Office (2014), 50-60% of the parents’ generation expected grandparents to talk and play with their children and pass on their experiences and wisdom to their children, whereas 40% of the parents’ generation expected grandparents to discipline them in daily life [28]. It has also been demonstrated that grandparents perceive themselves as role models and educators [18-20]. Grandparents play an essential role in the character development of grandchildren even in today’s era of nuclear families [30-36]. In this study, the family functions of grandparents were shown by daily care and healthcare and character formation of grandchildren. It was found that grandparents are involved in the child-rearing policy of the parent couple as emotional support for parents and think about whether the scope of helping the parent couple raise the child is excessive. According to Sumikawa (2016), grandparents state that it is the role of parents to raise their grandchildren and that they are involved in “not intervening too much in the childcare of their grandchildren as grandparents” [31]. Many grandparents tend to play a supportive but non-interfering role, set clear boundaries with the parents to avoid family conflict, respect parental wishes, and ensure family harmony and consistency in parenting [32-35]. The current study shows that emotional support for parents respects the child-rearing policy of parents and couples and protects their personal growth. To enhance the family function of grandparents, it is necessary to discuss in detail the division of roles between parents and grandparents, and midwives need to set up a discussion place for parents and grandparents. The role function of grandparents’ characteristics in this study was “own well-being. Grandparents expressed that raising their grandchildren was one of their purpose in life, whereas that the function of maintaining their own health was to maintain their own health. Grandparents experience a “rejuvenating effect” by being involved in childcare [37]. It was also revealed to result in better physical health [3]. Therefore, assessing the family functions related to the health balance of grandparents themselves will lead to support according to the needs of grandparents.

Factors Affecting the Family Functioning of Grandparents

The four family functions of grandparents were affected by the degree of involvement of grandchildren. Grandparents involved in raising grandchildren scored higher on four factors. Grandparents who did not feel burdened by taking care of their grandchildren recognized that raising their grandchildren helped them maintain their physical strength and youth. For grandparents, raising grandchildren has a positive impact on their health.

Regarding gender differences, grandfathers scored higher on the function of grandchildren’s character development and own well-being, whereas grandmothers scored higher on emotional support for parents. Further, grandmothers were more likely to raise their grandchildren for longer periods [38,39]. Grandparents’ employment status had no significant effect on their family functions. However, compared to those who were unemployed, grandparents working full-time perceived child-rearing as one of their reasons for living, whereas unemployed grandparents perceived that it reduced the time they could spend on work and hobbies. Grandparents who were unemployed held the belief that they respected the child-rearing policies of the parents [40] suggesting that further investigation is needed to determine whether grandparents’ employment influences their functions in the family. Grandparents’ age affected the dimensions of daily care and healthcare and emotional support for parents. Grandparents in their 40s and 50s were more involved in grandchild-rearing, recognizing that grandchild-rearing was one of their reasons for living and that grandchild-rearing was contributing to the family’s quality of life, even though it reduced their time available for work and hobbies [40]. It is assumed that grandparents, especially those who are younger, take on the role of caretakers while also having jobs and hobbies; we speculate that the balance between these two roles may affect grandparents’ family functions. It goes without saying that the closer the physical distance to the grandchildren, the more the family function of the grandparents. This study revealed that grandparents living with their grandchildren scored higher in daily care and healthcare and grandchildren’s character development than grandparents who did not live with them. Those who lived with their grandchildren had more intense parenting responsibilities and burdens, and the pressure to do a good job of raising their grandchildren often caused psychological stress, burnout, frustration, and feelings of being overwhelmed and helpless [19,41,42]. In contrast, grandparents who were only moderately involved in grandchild rearing, enjoyed the traditional laid-back grandparenting role without a sense of obligation to provide childcare [43]. The attitude of grandparents toward raising their grandchildren is that they consider it the responsibility of the parents to raise their grandchildren, but also to be involved when requested, and it is clear that they are responsible for raising their grandchildren in response to the parents’ requests [33,35,40]. Also, even if grandparents do not live with their children and grandchildren, they will watch over and adjust to them to maintain balance and live [44]. Therefore, in order to improve the quality of life of grandparents in their later years, it is necessary to provide support such as education and environment improvement so that grandparents can perform family functions, including maintaining an appropriate distance in preparation for welcoming new grandchildren.

Limitations of the Study and Future Research

This study clarified the family functions of grandparents with infant grandchildren. However, the family functions of grandfathers and grandmothers may be different in other scenarios. In some cases, grandparents responsible for grandchild rearing were at a higher risk of developing health problems and serious medical conditions, such as cardiovascular disease, arthritis, and generalized pain [45,46]. These topics should be explored in future research, as grandparents’ health status may affect their family functions. We also included grandparents with grandchildren aged 1 to 5 years and did not analyze their ages in detail. Grandparents have reported fatigue and lack of rest when caring for younger grandchildren [12,46]. In addition, differences in the choice of grandparents to assume the role of grandparents themselves or as obligations may affect the family function of grandparents [47]. Therefore, it is necessary to clarify the possibility that the burden of the role of grandparents is influenced by the recognition of whether or not they are responsible for raising their grandchildren as a duty.

Conclusions

The survey revealed grandparents’ family functions: daily care and healthcare, emotional support for parents, grandchildren’s character development, and own well-being. Grandparents who did not feel burdened reported that grandchild-rearing was one of their life passions and helped them maintain their own strength and youth. Grandparents’ involvement in raising their grandchildren affected their functions in the family. To enhance their family functions grandparents, it is necessary to assess the four functions of grandparents, support them in maintaining an appropriate distance from their grandchildren and children, and set up a forum for discussion between grandparents and parents.

Acknowledgments

We would like to thank Editage (http://www.editage.com) for English language editing.

Funding

This study was supported by Grant-in-Aid for Scientific Research Fundamental Research (C) under Grant 20K10896.

Competing Interests

The authors declare that they have no competing interests.

References

  1. Mori (2018) Current status of death during pregnancy and postpartum as seen from vital statistics (death, birth, stillbirth), perinatal medical care. Research on linkage database (Health, Labour and Welfare Sciences Research Grants, ICT Infrastructure Construction Research Project for Clinical Research).
  2. Ministry of Health, Labour and Welfare. Number of child abuse consultation cases at child guidance centers in FY29. 2018.
  3. Di Gessa G, Glaser K, Tinker A (2016) The health impact of intensive and nonintensive grandchild care in Europe: New evidence from SHARE. J Gerontol B Psychol Sci Soc Sci 71: 867-879. [crossref]
  4. Tahata J, Katane Y, Suzuki Y. Quality of life of grandmothers through childcare support [in Japanese]. J Womens Health 2012: 25-32.
  5. Kobayashi Y (2010) Fruits of returning home before and after childbirth Maternal Support and Mother-Child Relationship and Maternal Development. J Jpn Acd Midwif 24: 228-239.
  6. National Institute of Population and Social Security Research. 2015 Basic survey on social security and population problems [In Japanese]. 2017.
  7. Cabinet Office. The Reiwa 4th white paper on measures for a declining birthrate society [White paper] [In Japanese]. 2020.
  8. Yoda S, Shintani Y (2018) Mothers’ employment and childcare support from grandparents—An examination of intraindividual variation and interindividual differences [in Japanese]. of Population Problems. 74: 61-73.
  9. Di Gessa G, Glaser K, Price D, Ribe E, et al. (2016) What drives national differences in intensive grandparental childcare in Europe?. J Gerontol B Psychol Sci Soc Sci 71: 141-153. [crossref]
  10. Thomese F, Liefbroer A (2013) Child care and child births: The role of grandparents in the Netherlands. J Marriage Fam 75: 403-421.
  11. Kubo K, Oikawa Y, Katane Y (2011) Factor structure of grandmotherhood. Japanese Journal of Maternal Health 51: 601-608.
  12. Low SSH, Goh ECL (2015) Granny as nanny: Positive outcomes for grandparents providing childcare for dual-income families. Fact or myth? J Intergenerational Relatsh 13: 302-319.
  13. Sumikawa S (2009) Consideration of the introduction of grandparent classes for child-rearing support [In Japanese]. Japanese Journal of Maternal Health 50: 300-309.
  14. Ministry of Health, Labour and Welfare. Overview of the 2022 simplified life tables. 2022.
  15. Meyer MH, Kandic A (2017) Grandparenting in the United States. Innov Aging 1.
  16. ICM (2017) Definition of midwife, international definition of the midwife.
  17. Isoyama A, Shibuya E, Sakama, I (2016) Examination of the effectiveness of preparatory education programs for acquiring roles for grandparents who welcome new grandchildren Ibaraki Journal of Maternal Health. 35: 5-11.
  18. Lewis JP, Boyd K, Allen J, Rasmus S, et al. (2018) We raise our grandchildren as our own: Alaska native grandparents raising grandchildren in Southwest Alaska. J Cross-Cult Gerontol 33: 265-286. [crossref]
  19. Henderson TL, Dinh M, Morgan K, Lewis J (2017) Alaska native grandparents rearing grandchildren. J Fam Issues 38: 547-572.
  20. Zeng Z, Xie Y (2014) Effects of grandparents on children’s schooling: Evidence from rural China. Demography 51: 599-617. [crossref]
  21. Shorey S, Ng ED (2022) Socioecological model of parenting experiences: A systematic review. The Gerontologist 62: e193-e205. [crossref]
  22. Honohashi N (2008) Assessment method for family functions: Guide to FFFS Japan I. EDITEX. 10-11.
  23. Hangse T, Misawa S (2003) Creation of a family characteristic scale in Japan. Annals of Clinical Thanatology 8: 30-49.
  24. Soyama K, Yoshida K, Yoneda M (2015) Relationship between grandmothers’ child-rearing experiences and their awareness of raising grandchildren [In Japanese]. Journal of Japan Society of Nursing Research 38: 139-149.
  25. Tabuchi M, Nakahara J (2007) Motivation to support child-rearing and awareness of issues in the grandparents’ generation. Behavioral sciences of life, aging, sickness and death. 12: 12-22.
  26. Pei-Chun K, Hank K (2013) Grandparents caring for grandchildren in China and Korea: Findings from CHARLSs and KLoSA. J Gerontol B Psychol Sci Soc Sci 69: 646-651.
  27. Glaser K, Price D, Ribe Montserrat E, Di Gessa G, et al. (2013) Grandparenting in Europe: Family policy and grandparenting in providing childcare. Grandparents Plus.
  28. Cabinet Office. Fiscal 2013 report on the awareness survey on child-rearing in families and communities [In Japanese]. Cabinet office. 2014.
  29. Shiraishi E, Inoue R (2017) Literature review on the content and awareness of grandparents’ participation in raising grandchildren. Bulletin of the Faculty of Health Sciences, Tokai University 22: 67-74.
  30. Fukue S, Fukuoka K, Arai S (2020) Influence of past grandparent functions on psychological development and elderly image of university students. Kawasaki Medical Welfare Journal 30: 95-107.
  31. Sumikawa S (2016) Conflict between grandparents and parents focusing on roles concerning the first grandchild. Japanese Journal of Maternal Health 56: 531-538.
  32. Monserud MA (2010) Continuity and change in grandchildren’s closeness to grandparents: Consequences of changing intergenerational ties. Marriage Fam Rev 46: 366-388.
  33. StGeorge JM, Fletcher RJ (2014) Men’s experiences with grandfathers: A welcome surprise. Int J Aging Hum Dev 78: 351-378. [crossref]
  34. Villar F, Celdrán M, Triadó C (2012) Grandmothers offering regular auxiliary care for their grandchildren: An expression of generativity in later life? J Women Aging 24: 292–312. [crossref]
  35. Von Humboldt S, Monteiro A, Leal I (2018) How do older adults experience intergenerational relationships? Different cultures and ambivalent feelings. Educ Gerontol 44: 501-513.
  36. Xie H, Loy S, Caldwell LL, Robledo M (2018) A qualitative study of Latino grandparents’ involvement in and support for grandchildren’s leisure-time physical activity. Health Educ Behav 45: 781-789.
  37. Bordone V (2017) The youthful effect of childcare beyond grandparenthood. IIASA Working Paper No. WP-17-013.
  38. Martin-Baena D, Mayoral O (2019) Gender differences on the psychological well-being of early Spanish grandparents caring for grandchildren. J Psychiatry Behav Health Forecast 2.
  39. Zhao D, Zhou Z, Shen C, Ibrahim S, et al. (2021) Gender differences in depressive symptoms of rural Chinese grandparents caring for grandchildren. BMC Public Health 21.
  40. Isoyama A, Kinugawa A (2021) A study of factors concerning the involvement of grandparents with preschool grandchildren in raising their grandchildren. Proceedings of the Annual Meeting of the Japanese Society of Japan Nursing Sciences. 41.
  41. Fauziningtyas R, Indarwati R, Alfriani D, Haryanto J, et al. (2019) The experiences of grandparents raising grandchildren in Indonesia. Work Older People 23: 17-26.
  42. Kwon YS, Yoo EK (2016) The “twilight childcare” experiences of grandmothers. Information (Japan), 19: 5601-5606.
  43. Blundon A (2013) The role of grandparents. In A. Singh & M. Devine (Eds.), Rural transformation and Newfoundland and Labrador diaspora. Sense Publishers.
  44. Suzawa H (2012) Families acquiring the role of grandparents: Through the complementary coordination of grandchildren’s generations and parents’ generations. Annual reports of Graduate School of Humanities and Sciences. 27: 115-124.
  45. Byers LG, Bragg JE, Muñoz RT (2012) American Indian grand-families: Trauma and services. J Ethn Cult Divers 26: 204-216.
  46. Chang YT, Hayter M (2011) Surrogate mothers: Aboriginal grandmothers raising grandchildren in Taiwan. J Fam Nurs 17: 202-223. [crossref]
  47. Ando K (2020) Recent trends in the study of grandparent-grandchild relationships. Fam Relat Studies 39: 57-68.

Polynucleotides/Sodium Hyaluronate Ovules for Postmenopausal Vulvovaginal Atrophy and Other Vaginal Environment Disorders

DOI: 10.31038/AWHC.2023624

Abstract

Introduction: Vulvovaginal atrophy, as the most troublesome symptom complex within the more comprehensive clinical picture of Genitourinary Syndrome of Menopause, severely impacts the postmenopausal women’s quality of life, their self-respect as still attractive women, and possibly the women’s couple relationship. The same may happen at a younger age, with vaginal dryness as an occasional side effect of estro-progestin oral contraceptives, hormone-releasing vaginal rings, and other disorders leading to a deranged vaginal environment. Based on a sound rationale, a new medical device based on natural-origin polynucleotides and sodium hyaluronate formulated as vaginal ovuli may help over the long term with those frequent hassles of middle-aged and younger women.

Methods: Design: a survey investigation based on a questionnaire compiled by three investigators familiar with the new vaginal ovuli medical device in their everyday office practice. The investigators completed the survey based on outcomes in 45 ambulatory adult women (≥18 years old). All women had freely consulted the investigators to seek relief from vulvovaginal atrophy symptoms and vaginal dryness as a contraceptive side effect. The minimal inclusion and exclusion criteria helped simulate a real-world office situation. The survey followed a treatment cycle with a Class-III CE-marked medical device (Ovuli PNHA, Mastelli S.r.l., Sanremo, Italy—vaginal ovules with 0.25% polynucleotides, 0.25% non-cross-linked sodium hyaluronate, and 3% polycarbophil as functional ingredients). After the end of the self-administration cycle (one ovule intravaginally per day for two to four weeks), the survey’s goal was to assess the impressions of investigators and their patients about the safety and efficacy (symptom relief, restoration of a healthy vaginal environment) of the medical device in all conditions of vulvovaginal atrophy/vaginal dryness and dystrophic lesions/vaginal environment disorders. The device establishes a protective, moisturizing, and pH-controlled lubricating film onto the cervix and vaginal mucosa and promotes its physiological regenerative and reparative mechanisms. Assessment tools at baseline and after the end of the treatment cycle: Vaginal Health Index score (validated investigator-assessed five-item scale, with scores ranging from 5 to 25 and <15 as dryness and atrophy score threshold); subjective VVA symptom severity (vaginal dryness, vulvovaginal irritation/itching, vulvovaginal soreness, dyspareunia) assessed by the patients with the support of a series of five-point VAS scales (0=absent, 1=mild, 2=moderate, 3=severe, 4 = very severe).

Results: The surveyed investigators stated that the mean total VHI score significantly improved over the follow-up period, from 2.5 ± 0.94 to 3.6 ± 0.85 (p <0.001 vs. baseline). The mucosal thickness and integrity, showing severe or very severe atrophy in 13 surveyed women at baseline, presented no more than some mild atrophy in 91.1% of women, with 35.6% of surveyed women reverting to complete restitutio ad integrum. Vaginal dryness also markedly decreased from baseline, with 79% of cohort women improving to mild atrophy or symptom clearance and 21% reverting to restitutio ad integrum. Regarding the subjective symptom relief, the surveyed women reported a highly significant reduction of the mean total VAS score — cumulatively accounting for vaginal itching, vaginal burning, vaginal pain, pain/discomfort during intercourse, and vaginal dryness — from 3.5 ± 0.70 to 2.1 ± 0.73 (p <0.001 vs. baseline). The occasional mild local pain and irritation were of no clinical significance and rapidly transitory.

Conclusions: The novel formulation of polynucleotides and sodium hyaluronate as vaginal ovules efficiently counteract the symptom complex due to vulvovaginal atrophy in postmenopausal women, including younger women with contraindications for estrogen therapy and vaginal dryness as a contraceptive side effect. The study confirms that PNs, in synergy with hyaluronic acid, retain their innovative potential as non-pharmacological activators of physiological regenerative and reparative processes in vulvovaginal tissues. The study also confirms the novel device’s safety and the women’s compliance with treatment.

Keywords

Dyspareunia, Hyaluronic acid, PNHA ovules, Polynucleotides, Vaginal dryness, Vulvovaginal atrophy

Introduction

The role of 17ß-estradiol in so many cellular pathways regulating vulvovaginal cell growth, barrier functions, and cell proliferation and trophism explains why menopausal hypoestrogenism may have a grievous impact on middle-aged women [1]. Unfortunately, women in advanced countries may expect, on average, to live more than three decades in postmenopausal condition, and about 27% to 84% of them will experience the most troublesome vulvovaginal atrophy (VVA) symptoms [2]. Severe vulvar irritation and burning, compromised vaginal lubrication with discomfort or frank dyspareunia during sexual activity, vaginal discharge, dysuria, and recurrent urinary tract infections may thus be unwelcome companions for many years. The consequences for daily living activities are dire for 75% of symptomatic women, according to the outcomes of the “Vaginal Health: Insights, Views & Attitudes” (“VIVA”) online survey of 3,520 postmenopausal women in six countries [3-6].

Combined with hyaluronic acid (HA), highly purified DNA polynucleotides (PNs), purified from male salmon trout gonads, already showed to improve genital atrophy, VVA symptoms, and the disrupted sexual life and couple relationship of menopausal women [7-11]. PNs showed a powerful restructuring effect on local connective tissues after infiltration in the dermis and tonaca propria of vulvovaginal tissues [9,10]. PNs act non-pharmacologically by progressively releasing small nitrogen precursors (nitrogen bases, nucleosides, nucleotides) in vulvovaginal tissues and showing persistent moisturizing effects.7 Now, a new handy medical device formulation of natural-origin PNs and strongly hydrating HA as vaginal ovuli (formulation henceforth labeled PNHA) promises to extend their combined benefits to other troubling vaginal environment disorders. Examples are the younger women’s vaginal dryness and dyspareunia occasionally observed as adverse effects of estro-progestin oral contraceptives, birth control vaginal rings releasing hormones, and especially progesterone-only contraceptive methods [12-14].

The paper reports on the outcomes of a retrospective survey centered on a questionnaire prospectively administered by the investigators, all gynecologists, to some of their patients who spontaneously sought help and office treatment for problems of menopausal vulvovaginal atrophy (VVA). Some younger women sought relief for their vaginal dryness as a side effect of estro-progestin oral contraceptives or birth control hormone-releasing vaginal rings.

All investigators already used the investigated PNHA device as described in the approved product information leaflet (IFU)-one PNHA ovule daily [15]. The survey study, performed after two to four weeks of everyday use of the intravaginal medical device, was the first within a long-term monitoring program of the device’s persistent efficacy and profile of known side effects and contraindications. Identifying unknown side effects or emergent risks was another purpose of the study and the ongoing long-term monitoring program. Resorting to real-world data, independently from the rigid inclusion and exclusion criteria of randomized clinical studies with their highly selected patient samples, is the clue that supports the clinical value of the investigation in daily practice.

Methods

Study Design

Conceived as a single-arm ambulatory cohort investigation of adults of both genders who had spontaneously sought specialist help for postmenopausal vaginal environment problems and vaginal dryness as a side effect of estro-progestin oral contraceptives and birth control hormone-releasing vaginal rings. Before the survey, following the investigators’ prescription, all individuals had self-administered, at home in a real-world setting, the monitored Class-III CE-marked medical device (Ovuli PNHA, Mastelli S.r.l., Sanremo, Italy: vaginal ovules with 0.25% polynucleotides, 0.25% non-cross-linked sodium hyaluronate, and 3% polycarbophil as functional ingredients). Dose and self-administration period: one ovule intravaginally per day for two to four weeks as stated in the approved product information leaflet (IFU) [15].

After the last ovule domiciliary self-administration, the survey, completed by the investigators, was purely observational with no active intervention. All subjects agreed to submit to the survey after being informed about its goals. Beyond monitoring the efficacy and safety outcomes, the reasons for vaginal treatment were also registered. Questionnaires allow information collection without time constraints for the investigator to answer questions thoroughly, faithfully, and more quickly than face-to-face interviews.

The office-based survey study respected the Helsinki Declaration and Good Clinical Practice principles. All study materials, which included informed consent forms and questionnaires, were preliminarily peer-reviewed for ethical problems.

Observational Efficacy Assessments

Primary Efficacy Endpoint

Relieving objective signs and restoring the healthy vaginal state is paramount in managing vulvovaginal atrophy and disorders of the vaginal environment. The improvement of vaginal signs and symptoms from baseline, evaluated by the investigators with the help of the validated Vaginal Health Index, was the observational primary efficacy endpoint (assessment: total VHI score change from baseline after the last PNHA-based vaginal ovule). First described in 1995, the VHI is a five-point investigator-assessed scoring scale extensively used in clinical trials, which considers the lack of mucosal moisture, vaginal elasticity and volume of vaginal secretions, intravaginal pH, and the evidence of vaginal petechiae and bleeding (Table 1 [16,17]. The ranges of VHI subscale scores are 1 = none, 2 = poor, 3 = fair, 4 = good, and 5 = excellent, with the total VHI score between 5 and 25. Total VHI scores below 15 signal vaginal atrophy [15]. The unimodal symmetric distributions of outcomes on five-point Likert-like assessment tools like the VHI subscales minimize the statistical liability of skewed J- and U-shaped distributions; outcomes assessed on five-point scales also have lower means, floor, and ceiling effects. At the same time, the regression analysis shows that these scales explain a significant fraction of the variation in floor and ceiling effects while minimizing the contribution of unknown factors [16].

Table 1: Vaginal Health Index (Bachmann et al.); * Lower scores: progressively more severe atrophy [16]

Score

Overall elasticity *

Secretions type and consistency

pH

Mucosal epithelium

Moisture

1 None None 6.1 Petechiae noted before contact None; mucosa inflamed
2 Poor Scant, thin yellow 5.6−6.0 Bleeds with light contact None; mucosa not inflamed
3 Fair Superficial, thin white 5.1−5.5 Bleeds with scraping Minimal
4 Good Moderate, thin white 4.7−5.0 Not friable, thin mucosa Moderate
5 Excellent Normal (white flocculent) ≤4.6 Not friable, normal mucosa Normal

Secondary Efficacy Endpoint

Subjective assessment by patients of symptom severity (vaginal dryness, vulvovaginal irritation/itching, vulvovaginal soreness, dyspareunia) with the support of a series of impromptu five-score Visual Analog Scales (henceforth VAS, 0=absent, 1=mild, 2=moderate, 3=severe, 4 = very severe) [18].

Safety

Based on spontaneous reporting over the whole study period. The questionnaire included closed and open questions to identify known side effects, describe their clinical presentation, severity, and duration, and detect any previously unknown adverse event.

Statistics

The sample size estimation took advantage of the G*Power statistical program version 3.14 [19]. The estimate, after a literature review of the efficacy of comparable products, was based on a fictional cumulative VAS symptom score and a conservative assumption of the mean cumulative VAS symptom score at the end of the treatment course (60% improvement). Under these assumptions, the statistical power (two-tailed) to detect a significant difference in a treatment cohort of at least 42 women would have minimized the risk of false-negative type II errors (ß-risk = 0.92).

Descriptive statistics: tabulated as means ± standard errors of the mean (SEM). Inferential statistics: mean total VHI score and subscores as primary efficacy parameter and self-assessed symptom severity VAS scores as secondary efficacy parameter: non-parametric Wilcoxon test [18]. All statistical analyses, two-tailed with a 5% significance level, were performed with the StatPlus analysis program Version v7 [20].

Results

The three participating clinical investigators, all experienced gynecologists already prescribing the PNHA vaginal ovules, enrolled 45 ambulatory menopausal and non-menopausal women with VVA symptoms. All women completed the study fully compliant with the protocol and domiciliary vaginal ovule self-administration with only a few transitory and mild local adverse effects and without dropouts. Table 1 illustrates the women’s demographics, menopausal or non-menopausal conditions, and previous use of estrogens. The subgroup of menopausal women had been in menopause for a range of 2 to 30 years and had experienced VVA symptoms for periods ranging from 1 month to 20 years (Figure 1). Often together with oral or topical estrogens, 16 cohort women (35.6%) had used other topical and parenteral formulations and techniques to counter their VVA symptoms—other injectable polynucleotide and hyaluronic acid gels, hyaluronate topical formulations, vaginal moisturizers, lubricants, and electroporation and radiofrequency biostimulation procedures.

Table 1: Women’s demographics, menopausal condition, and oral and topical estrogen use. SEM: Standard Error of the Mean.

Age Mean ± SEM (years) 55.4 ± 10.29
Median (years) 56
Range (years) 25-74
Menopause Yes (%) 29 (64.4)
Years in menopause ± SEM 9.2± 6.64
Years with VVA symptoms ± SEM 6.6 ± 5.46
No (%) 16 (35.6)
Oral estrogens Yes (%) 7 (15.6)
No (%) 38 (84.4)
Topical estrogens Yes (%) 5 (11.1)
No (%) 46 (88.9)

FIG 1

Figure 1: Years with VVA atrophy symptoms in the 45-strong women’s cohort

At baseline, the VVA symptoms appeared to severely affect the women’s sexual life, with 31 menopausal and non-menopausal women (68.9%) reporting reduced sexual activity with decreased sexual interest and desire in 27 (60.0%).

The VVA signs and symptoms which most commonly induced the investigators to prescribe the PNHA intravaginal ovules were, in decreasing frequency, vaginal dryness (80% of cohort women), vaginal burning (40% of women), vaginal itching (37.8% of women), vaginal pain (31.1% of women), often during intercourse (24.4% of women), and vaginal bleeding during intercourse (15.6% of women). Symptoms co-existed very commonly. Previous genital surgery and dysuria were other significant reasons for prescribing the PNHA ovules (6.7% and 2.2% of women, respectively).

Efficacy

At baseline, the mean VHI score was 12.68, beyond the VVA threshold, while the mean total VHI score (mean of all VHI subscale scores) was 2.5 ± 0.94, once again describing a severe VVA picture. Figure 2 shows the highly significant improvement in the mean total VHI score at the study end compared with baseline, although always with some persisting atrophy signs and symptoms. Figure 3 analytically illustrates the improvements vs. baseline for each VHI subscore after treatment with the PNHA ovules.

FIG 2

Figure 2: Mean total Vaginal Health Index scores (± standard errors of the mean); **p<0.001 vs. baseline

FIG 3

Figure 3: Mean VHI subscores (± standard errors of the mean); **p<0.001 vs. baseline

Looking more in detail into the mucosal thickness and integrity (Figure 4), the ultimate basis for bleeding and other symptoms, thirteen women showed severe or very severe atrophy of the vaginal epithelium at baseline (scores: 1 or 2); after the topical PNHA treatment, 91.1% of women showed no more than a condition of mild atrophy (scores: 4 or 5), with 35.6% of them reverting to complete restitutio ad integrum (score: 5).

FIG 4

Figure 4: Percent of cohort women with compromised vaginal mucosal thickness and integrity of variable clinical severity (scores 1 to 5) at baseline and end-of-treatment assessment visits.

Regarding the clinical severity of vaginal dryness, the most bothersome and frequently reported symptom of atrophy, it also markedly decreased from baseline in all cohort women (Figure 5), with 79% of them improving to a condition of mild atrophy or symptom clearance (scores: 4 or 5), and 21% of them reverting to complete restitutio ad integrum (score: 5).

FIG 5

Figure 5: Percent of cohort women who reported vaginal dryness of variable clinical severity (scores 1 to 5) at baseline and end-of-treatment assessment visits.

Safety

The investigators and treated women invariably deemed the procedure manageable and handy without unexpected technical difficulties, troubles, and discomfort. The repeated insertion of the PNHA ovules was well tolerated, with a few transitory and mild local adverse effects—local burning in 15 cohort women (mild 31.8%, moderate 2.3%), vulvovaginal itching in 16 women (mild 31.8%, moderate 4.5%), and local discomfort in 19 women (mild 36.4%, moderate 6.8%). According to investigators, two women reported some mild vaginal blood losses, which disappeared in 6 to 10 days and were unrelated to the PNHA topical treatment. There were no unexpected technical difficulties. The few transitory and mild local adverse effects (local burning, vulvovaginal itching, and local discomfort) were of no clinical significance. They resolved spontaneously in a few hours without further therapy. There were no unexpected side events or complications (Figures 6 and 7).

FIG 6

Figure 6: Mean total symptom VAS scores (± standard errors of the mean); **p<0.001 vs. baseline

FIG 7

Figure 7: Mean VAS scores (± standard errors of the mean) for some representative symptoms; **p<0.001 vs. baseline.

Discussion

Real-world investigations aim to provide reliable insights into conditions analogous to everyday clinical practice [21]. Within a long-term monitoring program, the PNHA vaginal ovule formulation confirmed its efficacy and lack of unknown and troublesome side effects with once-daily dosing in menopausal VVA and other non-menopausal and post-surgical disorders with vaginal dryness and other symptoms, as stated in the Information for Use leaflet.

In this first step of the ongoing real-world, long-term monitoring program of the once-daily PNHA vaginal ovule medical device, the formulation proved to be effective with no intolerably bothersome side effects in vulvovaginal atrophy and other menopausal and non-menopausal or post-surgery disorders.

The phenotypic expression of postmenopausal vulvar involution — depletion of labia majora adiposity, blundering of interlabial sulci, loss of pigmentation and hair, reduced density of sweat and sebaceous production, preputial retraction with clitoral exposure and chronic irritation, and overall dysfunction of the vaginal ecosystem — and related symptoms are a burden on the woman’s self-confidence and self-image. Microscopically, the fragmentation and fusion of elastin fibers, collagen hyalinization, and extracellular matrix depletion are the markers of the VVA picture [2,22]. Moreover, vaginal dryness is a frequent side effect of estro-progestin oral contraceptives and hormone-releasing vaginal rings in up to 12.7% to 30.4% of women after three menstrual cycles, especially with preparations with the lowest synthetic estrogen content [23].

The hydrophilic polynucleotide polymers of the PNHA formulation reorganize in vulvovaginal tissues into a three-dimensional gel that binds water with a moisturizing and volume-increasing effect that synergizes with the potent HA hydrating effect [7-11]. Over the longer term, the polynucleotide component of the PNHA device passively replenishes the fibroblast pool of nitrogen bases, nucleosides, and nucleotide precursors and supports the dermal fibroblast viability, thus facilitating the production of new collagen fibers—the rationale for exploring the PNHA option to antagonize the menopausal and non-menopausal VVA and vaginal dryness [7-10].

The survey study had two main problems: compensating for the lack of a control group and the ß-risk of failing to detect a significant difference in semiquantitative scores, compared with baseline, at the end of the self-administration at home. However, the impressive efficacy outcomes reported by surveyed investigators and women are unlikely to be incidental findings and likely compensate for the first bias. Moreover, the cohort size of the enrolled women, more numerous than what was estimated to reduce to almost zero the ß-risk under the conservative assumption of a 60% efficacy, compensated for the second bias.

All surveyed clinical signs and symptoms showed highly significant improvements over the follow-up period, demonstrating that the newly introduced PNHA ovules are an effective option in all forms of vaginal atrophy, independently of cause. The clinically meaningful relief of VVA objective signs was not associated with more than some occasional undefined discomfort and mild burning of no clinical significance.

In conclusion, improving VVA-related objective signs means that the novel PNHA vaginal ovule medical device is an effective, safe, and well-tolerated non-hormonal therapeutic option in postmenopausal women with VVA and all women unwilling or unable to consider estrogen therapy. In general, the new PNHA device appears effective, independently of age, in all situations of vaginal dryness and a deranged vaginal environment for whatever cause.

References

  1. Cotreau MM, Chennathukuzhi VM, Harris HA, et al. (2007) A study of 17beta-estradiol-regulated genes in the vagina of postmenopausal women with vaginal atrophy. Maturitas 58: 366-376. [crossref]
  2. Nappi RE, Martini E, Cucinella L, et al. (2019) Addressing vulvovaginal atrophy (VVA)/Genitourinary Syndrome of Menopause (GSM) for healthy aging in women. Front Endocrinol (Lausanne) 10: 561. [crossref]
  3. NAMS Position Statement. The 2020 genitourinary syndrome of menopause position statement of The North American Menopause Society. Menopause 27: 976-992.
  4. Nappi RE, Davis SR (2012) The use of hormone therapy for the maintenance of urogynecological and sexual health post WHI. Climacteric 15: 267-274. [crossref]
  5. Davis SR, Lambrinoudaki I, Lumsden M, et al. (2015) Menopause Nat Rev Dis Primers 1: 15004.
  6. Shifren JL, Zincavage R, Cho EL, Magnavita A, Portman DJ, et al. (2018) Women’s experience of vulvovaginal symptoms associated with menopause. Menopause 26: 341-349. [crossref]
  7. Colangelo MT, Govoni P, Belletti S, Squadrito F, Guizzardi S, et al. (2021) Polynucleotide biogel enhances tissue repair, matrix deposition, and organization. J Biol Regul Homeost Agents 35: 355-362. [crossref]
  8. Bartoletti E, Cavallini M, Maioli L, et al. (2020) Introduction to Polynucleotides Highly Purified Technology. Aesthetic Medicine 6: 43-47.
  9. Palmieri IP, Raichi M (2019) Biorevitalization of postmenopausal labia majora, the polynucleotide/hyaluronic acid option. Obstet Gynecol Rep 3.
  10. Palmieri IP, Raichi M (2022) Vulvar rejuvenation with polynucleotides HPT® and benefits on postmenopausal sexual life disruption. Obstet Gynecol Rep 6. [crossref]
  11. Angelucci M, Frascani F, Franceschelli A, Lusi A, Garo ML (2022) Climacteric 25: 490-496.
  12. Sabatini R, Cagiano R (2006) Comparison profiles of cycle control, side effects and sexual satisfaction of three hormonal contraceptives. Contraception 74: 220-223. [crossref]
  13. Mares P, Hoffet M, Rousseau O, Ripart-Neveu S (2001) Vaginal dryness. Rev Prat 51: 155-158.
  14. Shah MB, Hoffstetter S (2010) Contraception and sexuality. Minerva Ginecol 62: 331-347. [crossref]
  15. Newest Gyn ovuli PNHA product information leaflet (IFU).
  16. Bachmann G (1995) Urogenital ageing: an old problem newly recognized. Maturitas S1-S5. [crossref]
  17. Cagnacci A, Xholli A, Sclauzero M, Venier M, Palma F, et al. (2019) Vaginal atrophy across the menopausal age: results from the ANGEL study. Climacteric 22: 85-89. [crossref]
  18. Ettinger B, Hait H, Reape KZ, Shu H (2008) Measuring symptom relief in studies of vaginal and vulvar atrophy: the most bothersome symptom approach. Menopause 15: 885-889. [crossref]
  19. Faul F, Erdfelder E, Buchner A, Lang AG (2009) Statistical power analyses using G*Power 3.1: Tests for correlation and regression analyses. Behav Res Methods 41: 1149-1160. [crossref]
  20. Statistical software downloaded from https://www.analystsoft.com/en/products/statplus/#statfeatures.
  21. Miksad RA, Abernethy AP (2018) Harnessing the power of real-world evidence (RWE): a checklist to ensure regulatory-grade data quality. Clinical Pharmacology & Therapeutics 103: 202-205. [crossref]
  22. Pérez-López FR, Phillips N, Vieira-Baptista P, Cohen-Sacher B, Fialho SCAV, et al. (2021) Management of postmenopausal vulvovaginal atrophy: recommendations of the International Society for the Study of Vulvovaginal Disease. Gynecol Endocrinol 37: 746-752. [crossref]
  23. Roumen FJM (2008) Review of the combined contraceptive vaginal ring, NuvaRing. Ther Clin Risk Manag 4: 441-451. [crossref]

Destructive Complicated Polyonychomycosis and Nail Incarnations: Peculiarities of Interventions, Author’s Views on Complex Treatment of Patients with Comorbid Background Pathology, Diabetes Mellitus

DOI: 10.31038/JCRM.2023621

Introduction

Surgical onychopathology includes a group of nosological forms of purulent-necrotic and mycotic-associated lesions of the nail phalanx, nail and peri-nail tissues, epidermophytia of the feet occupies one of the prominent places among pathological skin lesions; poses a risk of spreading to patients with diabetes due to high contagiousness, frequency of various complications; leads to a violation of the quality of life. Onychomycotic lesions are macroscopically classified into superficial mycosis, proximal and distal-lateral onychomycosis, onychocheilosis, subungual hyperkeratosis, and onychogryphosis. Pathological changes in the nail plate of the foot in patients with diabetes are often combined: hypertrophy and onychogryphotic deformation of the nail (in the form of an eagle’s or vulture’s beak) with its secondary growth, which is combined with the presence of dermatophytes (trichophytes, onychomatrichoma), pathological layering on the nail bed (brown color with destruction, disintegration) and the formation of multiple purulent bacterial-mycotic foci with abscessation. Complex treatment of patients with severe fungal lesions, complicated destructive onychomycosis with multiple nail lesions, onychogryphosis with incarnation, in particular in patients with diabetes, determines the removal of nail plates and the use of antifungal therapy, correction of background vascular pathology and is an urgent problem of outpatient surgery and dermatology. Some aspects of epidemiology, etiology, pathogenesis of onycheal and subungual pathology, macroscopic types of lesions, risk factors and cases of nail incarnations – ingrown nail (IN), localization, frequency of occurrence and causes of purulent onychia, paronychia, other complications and recurrences. In our previous publications, the effectiveness of various methods of surgical interventions, isolated and in combination with other methods of complex treatment, features of the postoperative period in isolated and combined lesions, in particular complicated destructive polyonychomycosis (UDP), were analyzed. It has been established that conservative and orthopedic treatment of IN and other mycotic surgical lesions of nails are not very effective, while the main methods are Dupuytren’s nail removal, Emmert-Schmiden operation, etc. in 2-20% of cases (depending on the absence or presence of onychocryptosis and associated fungal lesions) cause relapses. Complicated mycotic lesions and HF in patients with diabetes are a special problem.

The goal of the work is to improve the implementation of interventions, comprehensive treatment of patients with complicated polyonychomycosis with comorbid background pathology, diabetes and to implement an original method of elimination of UDP – hyperkeratosis and onychogryphosis with nail incarnation – secondary nail growth, in particular in patients with diabetes to optimize early and long-term results.

Materials and Methods

In the 4th City Clinical Hospital of Lviv and MC “Salyutas”, the prospective material for a 10-year period was 496 cases of destructive onychomycosis complicated by secondary nail incarnation, of which 320 were men and 176 were women. The age of the operated patients is from 35 to 72 years. The main group consisted of 325 patients – 172 men and 153 women aged 43-68 years. The control group included 171 cases of onychodestruction that were treated according to “classical” schemes. All subsamples were statistically similar and comparable in terms of age and gender composition. Imperfect surgical techniques account for more than half of the causes of relapses, including cases of IN with subungual hyperkeratosis or onychogryphosis, χ2=20.13, p=0.01. The highest frequency of repeated INs among the technical reasons for recurrence was observed in unjustified refusal of partial matrixectomy – 19.8% and with traumatic mobilization/removal of nails (traumatic onychectomy). The hallux of the left foot was most often affected – in 48.85%, the right – in 33.26%, the presence of IN of both hallux was found in 17.89% of patients. In 18.8% of the sample, onychocryptosis occurred against the background of obliterating diseases of the arteries of the lower extremities: atherosclerosis in 13.76% and diabetes in 15.05%. All surgical interventions in outpatient conditions were performed under adequate anesthesia. The Mann-Whitney U-test, χ2-test (V-square), χ2-test with Yates’ correction for continuity, Fisher’s exact test (Fisher exact p). We determined the χ2 degree of influence of the studied phenomenon (etiological factor, morphogenetic mechanism) on the development of nail plate incarnation and other complications. “Classical methods” of descriptive statistics were also used, with a confidence interval of 95%, calculations were performed on licensed software. The study meets the requirements of bioethics.

Results and Discussion

We have researched clinical variants and peculiarities of the course of the pathological process, confirmed the optimal methods of surgical intervention, their phasing and the terms of complex treatment of complicated, combined mycotic processes and recurrences of reincarnations of nail plates and lesions of the nail structures. Adjuvant systemic three-day antimycotic pulse therapy with daily intake of 400 mg of itraconazole was applied for three days prior to interventional/operative treatment. The peak frequency of mycotic lesions with HF occurred in the subsamples of people aged 40-50 years and in people aged 50-60 years. Pathological incarnations of mycotically altered nail plates were characteristic of trichophytosis in elderly patients. It has been claimed that the free lateral edges of the nail with UDP due to the lifting of its central part by hyperkeratosis are “undermined”, compressing perieponycheal tissues and eponychia. We used the following methods of surgical treatment. “Classic” removal of VN (Dupuytren’s operation) was performed in 64 people (12.90%), en bloc eponychectomy (excision of pathologically changed eponycheal tissues) + nail removal through onycholised structures (low-traumatic) + dermatophytoma resection + partial matrixectomy + eponycheoplasty – in 57 others cases (11.49%), nail removal through onycholised structures (low-traumatic) – in 51 (10.28%), “classic” nail removal + excision of pathologically changed eponycheal tissues – in 43 (8.66%), en bloc eponyhectomy ( excision of pathologically changed eponycheal tissues) + nail removal through onycholated structures (low-traumatic) + resection of dermatophytoma + partial matrixectomy – in 40 (8.04%) block-like eponychectomy + nail removal through onycholised structures (less traumatic) + partial matrixectomy – in other 8 (1.61%) patients. Thus, removal of ingrown nails was used in all cases, of which 64 people (12.90%) had Dupuytren’s nail removal without anti-relapse supplements, extended ablation with excision of dermatophytoma – in other 57 cases (11.49%). In the vast majority of patients (87.1% of the sample), surgical treatment of UDP with VN with anti-recurrence components was applied – two- or three-component, supplemented by excision of pathologically changed eponycheal tissues or (and) mechanical abrasion or (and) coagulation partial matrixectomy. Patients with diabetes accounted for 17% of the sample and were adequately represented in the main and control groups and in the clinical subsamples.

In patients with type 2 diabetes, probable strong positive correlations were recorded between all indicators of carbohydrate metabolism: glucose with insulin (r=0.52; p<0.01), with the NOMA index (r=0.69; p<0.01), with glycosylated hemoglobin (r=0.76; p<0.001); insulin with NOMA index (r=0.74; p<0.01) and glycosylated hemoglobin (r=0.65; p<0.01); NOMA index with glycosylated hemoglobin (r=0.69; p<0.01). We proposed and implemented a method of removing affected nail plates in patients with UDP, in particular, in the presence of trichophytic nail hyperkeratosis, polyonychogryphosis. This method is applied and has proven itself positively in diabetic patients with purulent / incarnate lesions associated primarily with Tr. Rubrum infection.

After applying a tourniquet to the base of the finger, preparing the operating field and anesthesia, stepping back from the proximal edge of the onychogryphous nail by 2-3 mm, we perform a linear dissection of the soft tissues medially retronycheally to the nail plate, which is extended to the medialeponycheal ridge, cutting out pathologically changed medial eponycheal tissues in a block-like manner. We continue the medial dissection distally and linearly down 5 mm on the finger bundle. The remains of the medial eponycheal tissues are separated from the medial edge of the onychogryphous nail with a pedicure spatula for an ingrown onychogryphous nail, a blade of a sterile pedicure tool PE-60/1 (an inclined manicure file with a blade). We visualize the medial edge of the onychogryphous nail and conduct a visual macroscopic assessment of it for the presence of deformations, delaminations, indentations, foci of hyperkeratosis. Similarly, retreating from the edge of the onychogryphous nail by 2-3 mm, we perform a lateral retronycheal linear dissection of the soft tissues to the nail plate, which we extend tolateral eponycheal ridge, cutting out pathologically changed lateral eponycheal tissues en bloc. We continue the lateral dissection distally and linearly down 5 mm on the finger bundle. The remains of the lateral eponycheal tissues are separated from the lateral edge of the onychogryphous nail with a pedicure spatula for an ingrown onychogryphous nail, the blade of a sterile PE-60/1 pedicure tool (an inclined manicure file with a blade). We visualize the incarnated lateral edge of the onychogryphous nail and conduct a visual macroscopic assessment of it for the presence of deformations, delaminations, serrations, foci of hyperkeratosis. Under the distal corner of the onychogryphous nail, in the area of the smaller incarnation, through the onycholized structures between the nail plate and the nail bed in the proximal-lateral direction, we insert the ax-shaped tip of the sterile nail blade PE-10/2 (with a rounded pusher with an ax-shaped raspator), the sterile pedicure tool PE-60/1, with which we gradually mobilize the nail plate with pendulum-like movements to the germinal zone and the proximal edge of the onychogryphous nail, gradually removing hyperkeratoid masses by scraping with a sterile PE-10/2 manicure spatula and a small Volkmann spoon, mobilizing and lifting the nail plate. We insert the blade of a sterile PE-30/4 manicure tool (a rounded pusher with a bent blade) into the formed channel, which completes the mobilization with pendulum-like movements. We introduce the straight blade of a PE-30 sterile manicure tool (a rounded pusher with a straight blade) with which in the proximal-contralateral direction we finally separate the mycotically changed thickened incarnate nail plate en block with the main mass of hyperkeratotic masses, grab it with a sterile Kocher clamp and remove it. With the ax-shaped tip of the sterile manicure blade PE-10/2 (rounded pusher with ax-shaped rasp) and the bent blade of the sterile manicure tool PE-30/4 (rounded pusher with bent blade), we gradually isolate from the eponycheal canals and the nail bed and mobilize in the distal direction the remnants of hyperkeratoids masses and dermatophytomas, which are also captured en block with a sterile Kocher clamp and removed. We clean the nail bed and eponycheal canals from the remnants of exfoliated epidermal structures. We clean the sinuses and eponycheal canals with a Volkmann spoon and a sterile pusher. We perform a revision of the medial and lateral edges of the wound for the tactile detection of the remaining fragments of the jagged edges of the onychogryphous nail. After devulsion of the remnants of eponycheal tissues in the area of incarnation of the onychogryphous nail with the bent blade of a sterile manicure tool PE-30/4, the remaining incarnate fragments of the nail plate are captured under visual and tactile control and removed with a Mosquito-type clamp. we perform bilateral partial marginal matrixectomy with longitudinal mechanical excision and diathermocoagulation of the germ zone and matrix of the onychogryphous nail in the areas of incarnations. Coagulated detritus is cleaned by scraping with the tip of a PE-60 sterile pedicure paddle. The sinuses, eponycheal canals and nail beds are cleaned with a Volkmann spoon and a sterile pusher. We clean the wound three times with a 3% solution of hydrogen peroxide and an aqueous solution of povidone iodine. After repeated devulsion of the remains of the eponycheal tissues with the blade of the PE-60/1 sterile pedicure tool, the eponycheal channels are filled with tampons made of iodoform gauze, keeping the remains of the eponycheal tissues removed. The central part of the wound is filled with povidone-iodine liniment, tightly tamped with gauze swabs under visual control. We remove the tourniquet from the base of the finger. Apply an aseptic bandage. We clean the wound three times with a 3% solution of hydrogen peroxide and an aqueous solution of povidone iodine. After repeated devulsion of the remains of the eponycheal tissues with the blade of the PE-60/1 sterile pedicure tool, the eponycheal channels are filled with tampons made of iodoform gauze, keeping the remains of the eponycheal tissues removed. The central part of the wound is filled with povidone-iodine liniment, tightly tamped with gauze swabs under visual control. We remove the tourniquet from the base of the finger. Apply an aseptic bandage. We clean the wound three times with a 3% solution of hydrogen peroxide and an aqueous solution of povidone iodine. After repeated devulsion of the remains of the eponycheal tissues with the blade of the PE-60/1 sterile pedicure tool, the eponycheal channels are filled with tampons made of iodoform gauze, keeping the remains of the eponycheal tissues removed. The central part of the wound is filled with povidone-iodine liniment, tightly tamped with gauze swabs under visual control. We remove the tourniquet from the base of the finger. Apply an aseptic bandage. tightly tampon with gauze tampons under visual control. We remove the tourniquet from the base of the finger. Apply an aseptic bandage. tightly tampon with gauze tampons under visual control. We remove the tourniquet from the base of the finger. Apply an aseptic bandage.

In the presence of polyonychomycosis with damage to more than 4 nail plates and damage to other structures of the foot (hand), surgical rehabilitation is divided into several stages. No more than four nail plates were removed simultaneously. In the case of other mycotic-associated pathology, no more than two simultaneous interventions were performed at the same time and no more than three nail plates were removed. The sequence of surgical interventions was selected according to their urgency. Nail plates affected by subungual hyperkeratosis were mobilized from the nail bed through hyperkeratoid masses and onycholated structures by a blunt approach to the retronycheal (posterior nail) ridge with fixation by the distal edge of the secondary VN, onychectomy was performed. A bare nail bed with remnants of hyperkeratoid layers and dermatophytomas in the distal part and hyperkeratoses near the eponycheal structures was visualized, which were removed by scraping, additional sanitation with a Volkmann spoon. Antimycotic pulse therapy was continued for the next 4 days of the postoperative period with daily intake of 400 mg of itraconazole. The remaining mycotically changed areas of the nails are cleaned daily with ciclopirox-containing antimycotic varnishes in order to prevent re- or mixed infection, as well as to prevent further fragmentation of the nails. In addition, three 7-day courses of pulse therapy with daily intake of 400 mg of itraconazole with a 10-day break between them were applied. In patients with existing obliterating lesions of the arteries of the lower extremities and in patients with diabetes, planned stages of surgical rehabilitation were performed only after complete epithelization under the guise of a course of vasodilator therapy. Dressings were performed every other day with treatment of surgical wounds with povidone iodine solution (until complete epithelization of wounds) and application of terbinafine liniment (until complete regrowth of nails) with sanitation of other remaining (unremoved) nails by applying antimycotic varnish every other day for 3 months. The last anti-recurrence course of UDP pulse therapy with itraconazole was carried out after the complete surgical rehabilitation of VN and polyonychomycosis. In patients with UDP and VN of the main group, in particular, in 57 patients with diabetes, in which the nail plates were removed due to onycholised structures, the healing time (crushing) of surgical wounds was 11-20 days (the average duration of healing – 16 days), in the control group – 15-25 days (the average duration of healing – 21 days); that is, in patients with UDP who underwent onychectomy due to onycholised structures, the healing time of onychectomy wounds was shorter. Good effectiveness of complex treatment was established in 284 patients of the main group (87.38%) and in 141 cases of the control group (82.46%). In general, a positive effect from the use of combined therapy with itraconazole and staged surgical removal of mycotically affected nails was confirmed in 425 patients (85.69%). who underwent onychectomy through onycholated structures, the healing time of onychectomy wounds was shorter. Good effectiveness of complex treatment was established in 284 patients of the main group (87.38%) and in 141 cases of the control group (82.46%). In general, a positive effect from the use of combined therapy with itraconazole and staged surgical removal of mycotically affected nails was confirmed in 425 patients (85.69%). who underwent onychectomy through onycholated structures, the healing time of onychectomy wounds was shorter. Good effectiveness of complex treatment was established in 284 patients of the main group (87.38%) and in 141 cases of the control group (82.46%). In general, a positive effect from the use of combined therapy with itraconazole and staged surgical removal of mycotically affected nails was confirmed in 425 patients (85.69%). In patients on the background of adjuvant systemic antimycotic treatment with daily pulse therapy of 400 mg of itraconazole, operative treatment of UDP with VN through onycholated structures with the use of podological pedicure instruments, supplemented by excision of pathologically changed eponycheal tissues, mechanical and coagulation partial matrixectomy, was applied to patients, this method can be recommended for the elimination of affected nails in patients with diabetes and obliterating lesions of the vessels of the lower extremities, as well as in patients with other vascular and/or neurotrophic pathology, elderly patients. The applied treatment scheme made it possible to achieve clinical and mycological recovery in 83.33% of the subsample. The achieved good result of the treatment was primarily determined by the peculiarity of the surgical approach, χ2=48.25, p<0.01.

The presence of foci of onycholysis and disintegration of areas of hyperkeratosis, which leads to detachment of part of the nail plate (χ2=15.23, p=0.0211), especially in elderly patients with background endocrine pathology – diabetes, justifies the feasibility of performing a minimally traumatic onychectomy. Patients with onychomycosis associated with secondary incarnation of the nail have a total hypertrophic fungal lesion with the formation of subungual hyperkeratosis or onychogryphosis (χ2=20.41, p=0.01) and subonycheal dermatophytoma, which complicates the mobilization and surgical removal of affected nails. Good effectiveness of the proposed treatment was established in 284 patients of the main group (87.38%) and in 141 cases of the control group (82.46%). In general, a positive effect from the removal of mycotically affected nails was confirmed in 425 patients (85.69%).

The Obtained Research Data Prove That

The author’s developed methods of surgical interventions, based on low-traumatic mobilization and resection/removal of affected nails through onycholated structures, determine a decrease in intraoperative damage to the nail bed (χ2=20.13, p=0.01), reduce the risk of mycotic contamination of other adjacent structures (χ2=27.41, p=0.01); are characterized by a decrease in the intensity of pain (χ2=48.32, p=0.01), a statistically significant increase in the speed of wound healing, the Popova index (χ2=32.14, p=0.01), an improvement in the results of complex treatment and the quality of life of patients. In the postoperative period, we apply dressings with antiseptics, antifungal (antimycotic) ointments and varnishes until the nail(s) grow back completely, according to indications – adequate systemic antifungal therapy. Partial marginal matrixectomy ensures the absence of growth of the nail plate in the area of resection, narrowing the nail and preventing its re-incarnation. Adequate classification criteria of purulent-necrotic complications of onychomycosis are proposed with the selection of the main types of lesions, in each of which clinical variants are differentiated according to the severity of morphological changes, which are of practical importance for the sequence of removal of affected nails. It is claimed that the use of minimally invasive nail removal helps to accelerate the elimination of the fungus and the healing of wounds, improve the results of complex treatment and the quality of life of patients. Adequate classification criteria of purulent-necrotic complications of onychomycosis are proposed with the selection of the main types of lesions, in each of which clinical variants are differentiated according to the severity of morphological changes, which are of practical importance for the sequence of removal of affected nails. It is claimed that the use of minimally invasive nail removal helps to accelerate the elimination of the fungus and the healing of wounds, improve the results of complex treatment and the quality of life of patients. Adequate classification criteria of purulent-necrotic complications of onychomycosis are proposed with the selection of the main types of lesions, in each of which clinical variants are differentiated according to the severity of morphological changes, which are of practical importance for the sequence of removal of affected nails. It is claimed that the use of minimally invasive nail removal helps to accelerate the elimination of the fungus and the healing of wounds, improve the results of complex treatment and the quality of life of patients [1-14].

Conclusions

For patients with destructive complicated polyonychomycosis, in particular, in the presence of secondary growths against the background of comorbid pathology, in particular, background diabetes, we have developed original methods of surgical interventions based on minimally traumatic mobilization and resection/removal of affected nails through onycholated structures, which determine the reduction of intraoperative damage to the nail bed, (χ2=20.13, p=0.01), reduce the risk of mycotic contamination of other adjacent structures (χ2=27.41, p=0.01); are characterized by a decrease in the intensity of pain (χ2=48.32, p=0.01), a statistically significant increase in the speed of wound healing, the Popova index (χ2=32.14, p=0.01), an improvement in the results of complex treatment and the quality of life of patients. In patients with incarnate onychomycosis, there is the formation of subungual hyperkeratosis or onychogryphosis and dermatophytoma, which complicates the mobilization and surgical removal of affected nails. In our clinic, in the complex treatment of complicated and combined fungal onychodestructions, we use minimally traumatic removal of nail plates in their destructive onychomycotic lesions. We remove the affected nail through onycholised structures using podological pedicure instruments, if there is an ingrowth, we supplement it with other local interventions – excision of pathologically changed eponycheal tissues, mechanical and coagulation partial marginal matrixectomy. In the postoperative period, we apply bandages with antiseptics, antifungal (antimycotic) ointments and varnishes until the nail(s) grow back completely.

References

  1. Cabete J, Galhardas C, Apetato M, Lestre S (2015) Onychomycosis in patients with chronic leg ulcer and toenail abnormalities. An Bras Dermatol 90: 136-139. [crossref]
  2. Chiu HH, et al. (2016) Onychomadesis following hand-foot-and-mouth disease. Cutis 97: 20-21. [crossref]
  3. Erdogan FG, Guven M, Erdogan BD, Gurler A (2014) Previous nail surgery is a risk factor for recurrence of ingrown nails. Dermatol Surg 10: 1152-1154. [crossref]
  4. Jung DJ, Kim JH, Lee HY, Kim DC, Lee SI, et al. (2015) Anatomical characteristics and surgical treatments of pincer nail deformity. Arch Plast Surg 42: 207-213. [crossref]
  5. Kallis P, Tosti A (2016) Onychomycosis and Onychomatricoma. Skin Appendage Disord 1: 209-212. [crossref]
  6. Kim JY (2012) Matrixplasty for the treatment of severe incurved toenail with growth plate deformity. J Am Podiatr Med Assoc 102: 198-204. [crossref]
  7. Leeyaphan C, Bunyaratavej S, Prasertworanun N, Muanprasart C, Matthapan L, et al. (2016) Dermatophytoma: An under-recognized condition. Indian J Dermatol Venereol Leprol 82: 188-189. [crossref]
  8. Nadashkevitch O, et al. (2017) Complicated onychomycosis and ingrown nail: complex treatment (case series). Proc Shevchenko Sci Soc Medical sciences 2: 64-68.
  9. Tucker JR (2015) Nail Deformities and Injuries. Prim Care 42: 677-691.
  10. Vergun, et al. (2017) Chronic mycotic-assotiated surgical nail pathology complicated with ingrown nail (nail incarnation): the analyses of clinical cases and complex treatment. International Journal of Medicine and Medical Research 3: 33-40.
  11. Vergun AR, et al. (2020) Surgical Incarnated Nail Pathology, Author Views of Some Controversion in Complex Treatment. Acta Scientific Medical Sciences 4: 31-37.
  12. Vlahovic TC (2016) Onychomycosis: Evaluation, Treatment Options, Managing Recurrence, and Patient Outcomes. Clin Podiatr Med Surg 33: 305-318. [crossref]
  13. Zeng M, Fu SP, (2012) Meliorated surgical procedure of Winograd for recurrent onychocryptosis. Zhonghua Yi Xue Za Zhi 92: 1767-1769. [crossref]
  14. Zhu X, Shi H, Zhang L, Gu Y (2012) Lateral fold and partial nail bed excision for the treatment of recurrent ingrown toenails. J Clin Exp Med 5: 257-261. [crossref]

Effects of Selected Primers and Vanishes on Open Dentine Tubules: An SEM Study

DOI: 10.31038/JDMR.2023612

Abstract

Dentine sensitivity (DS) is a common painful condition affecting teeth. The exact mechanism of transmission of an environmental stimulus across dentine is not fully understood; currently the most accepted theory is the hydrodynamic theory as proposed initially by Brannstrom. Treatment has been concentrated on either reducing the dentine fluid flow by occlusion of tubule openings, or altering the pulpal sensory nerve activity preventing transmission of pain to the central nervous system.

Aims and Methods: The aims of the present in vitro study were to examine the dentine tubule occluding and penetrating properties of selected in-office desensitizing agents (varnishes and primers) using scanning electron microscopy and a dentine disc model.

Results: Of the products examined, the fluoride varnishes, Bifluoride 12 and Duraphat, and Cervitec (a chlorhexidine containing varnish) were effective in both occluding and penetrating the dentine tubules. The results from All Bond 2, One Step and Scotchbond primers were superior to those of HEMA group primers, Gluma 3, Gluma CPS and Solobond Plus.

Conclusions: These findings suggest a mechanism for the action of these potential desensitising agents and suggest that the tubule penetrating properties may play a role in the longevity of their retention on the tooth. Investigation of surface coverage and tubule penetrating characteristics are both necessary in order to fully evaluate in vitro the desensitising potential of agents claimed to reduce Dentine Sensitivity.

Introduction

Dentine sensitivity (DS) can be defined as a pain arising from exposed dentine typically in response to chemical, thermal, tactile, or osmotic stimuli, which cannot be explained as arising from any other form of dental defect or pathology [1]. Currently, the most accepted theory of stimulus transmission across dentine is the hydrodynamic theory [2] proposed initially by Brannstrom [3]. According to this theory, minute fluid shifts across dentine in either direction in response to thermal, tactile, chemical or osmotic stimuli can stimulate mechanoreceptors in or near the pulp, which in turn excite the pulp sensory nerves to cause pain.

Pashley [4] reported that there are two approaches in the treatment of DS, 1) partial or complete occlusion of the dentine tubules and 2) alteration of pulpal sensory nerve activity (SNA) at or near the pulpo-dentinal surface. If the hydrodynamic theory of intradental nerve stimulation is accepted then the treatment of DS through tubule occlusion is a feasible and reasonable approach.

Clinically, DS has been treated by numerous agents, in-office and over the counter (OTC), which have claimed to effectively reduce pain arising from exposed dentine. Laboratory evaluation of these desensitizing agents using the dentine disc model has been reported in several studies [5,6], although, Mordan et al. [7] have modified this model to establish a more precise methodology control to evaluate potential desensitizing agents.

The aims of the present study were to investigate the degree of dentine surface coverage and the extent of tubule occlusion and penetration of selected in-office varnishes and primers. A dentine disc model and qualitative scanning electron microscopy (SEM) were used.

Methods and Materials

Surgically extracted, carious free, unerupted third molars were fixed in 3% glutaraldehyde in 0.1M sodium cacodylate buffer (CAB) solution (pH 7,4) at 4°C for up to one week. 1mm thick dentine discs were obtained from the region below the crown and above the pulp using a Testbourne diamond saw and stored in 0.1 M CAB at 4°C until required.

Before use, the dentine discs were ultrasonicated for 30 seconds in distilled water to dislodge cutting debris. The smear layer was then removed using either 6% citric acid for two minutes or the appropriate etchant provided by the manufacturer and applied according to their instructions. The discs were rinsed in distilled water and marked on either side for orientation (Figure 1) before being broken into halves using dental pliers to provide control and test sections [7].

FIG 1

Figure 1: Marking of dentine discs used in the study

The selected desensitising agents and their ingredients, according to the manufacturer’s information, are shown in Tables 1a and 1b. During the study all the products were coded and at least two discs were used for each agent. The selected desensitising agents were applied onto the test halves according to manufacturer’s instructions. The test and control halves were then allowed to dry in a desiccator for at least 24 hours.

Table 1a: Formulation and manufacturer information of test agents: Varnishes

Test agent

Manufacturer

Active ingredients

Bifluoride 12 Voco GMBH, 27457, Cuxhaven, Germany Fluoride varnish made of synthetic resin. Sodium and calcium fluoride
Cervitec Vivadent Ets. Shaan

Liechtenstein, Germany

1 g contains: 0.010 g chlorhexidine, 0.010 g thymol Polyvinyl Butyral (varnish) Ethanol, ethyl acetate
Duraphat Previously Rhone-Poulenc Rover GMBH, Nattermannallee 150829, Koln, Germany (Current Manufacturer Colgate Palmolive Company, USA) An alcoholic suspension of natural resins containing 5% NaF

Table 1b: Formulation and manufacturer information of test agents: Primers

Test agent

 Manufacturer

 Active ingredients

Hema-Seal G Germipherne Corporation

Ontario, Canada

HEMA, Glutaraldehyde, Sodium fluoride, Water
Gluma 3 Bayer Dental

Leverkusen, Germany

Aqueous solution of gluteral (Glutaraldehyde), HEMA
Gluma CPS Bayer Dental

Leverkusen, Germany

36,1% HEMA, 5,1% Glutaraldehyde, 58,8% Water
Solobond Plus

Universal Bonding Agent

Voco,

Cuxhaven, Germany

Methacrylate Acetone
All Bond 2

Universal Adhesive System

Dual cured

Bisco Inc.

Itasca, IL, 60143

Illinois, USA

N-phenylglycine-glycine methacrylate and bisphenyl dimethacrylate.
Scotchbond

Multi-purposed

Dental Adhesive System

3M Dental Products

Saint Paul, MN 55144

1000, USA

HEMA and BIS-GMA
One Step

Universal Dental Adhesive System

Bisco Inc.

Itasca, IL 60143

Illinois, USA

BPDM monomer dissolved in an acetone solution

A separate series of discs was prepared following the protocol described above, but both control and test halves were carefully fractured into quarters providing longitudinal surfaces for examination of the tubule lumen contents.

After drying in the desiccator, the discs were attached to SEM stubs and sputter coated using a Polaron E5000 sputter coater (Polaron U.K.) with a layer of gold/palladium. The specimens were viewed in a FEI/Philips XL30 FEG SEM (FEI, Eindhoven, Netherlands) at a working distance of 10 mm.

Micrographs were taken from selected fields in the central portion of each half disc on either side of the fractured edge and the test surfaces were only compared with the corresponding controls. Comparison of the test products’ ability to block and penetrate the dentinal tubules was also subjectively assessed using the micrographs.

Results

An example of a control disc with both surface and fractured profiles can be observed in Figure 2a showing open dentinal tubules in both views (surface and fractured).

FIG 2A

Figure 2a: Control Surface and Fracture

Varnishes

After application to the dentine disc, Cervitec (containing chlorhexidine) was observed to form a uniform layer that covered the whole dentine surface (Figure 2b). No tubule orifices, however were apparent. Fracturing the disc revealed the presence of a thick, textured layer, which covered the tubule orifices. There was some penetration of the product into the tubule lumen (arrows). Duraphat (containing fluoride) provided an uneven crystalline layer that covered the dentine surface and left tubule orifices visible (Figure 2c). Upon fracturing the dentine disc, a thin, amorphous surface layer was evident, although the varnish was usually present within the tubules, where penetration occurred it appeared to coat the lumen occupying the whole tubule diameter. When Bifluoride 12 (containing fluoride) was applied to the dentine disc, the surface was observed to be covered with small, irregular crystal-like structures and the tubule orifices were covered (Figure 2d). After fracturing the disc a thick, rough layer was observed which occluded the tubule orifices and occluded the tubules with plugs that occupied most of the tubule lumen (arrow).

FIG 2B

Figure 2b: Cervitec varnish Surface and Fracture views

FIG 2C

Figure 2c: Duraphat Varnish (Surface and Fractured views)

FIG 2D

Figure 2d: BiFluoride 12 Varnish (Surface and Fractured views)

Primers

The Gluma group of primers (Gluma 3 and Gluma CPS) as well as Hema-Seal G and Solobond Plus primer, whose principal active ingredients are glutaraldehyde and 2- hydroxyethylmethacrylate (HEMA), appeared to produce similar deposits on the dentine surface, partially occluding the tubule openings (Figures 3a-3f).

FIG 3A

Figure 3a: Hema-seal G Primer (Surface and Fractured views)

FIG 3B

Figure 3b: Gluma 3 Primer (Surface and Fractured views)

FIG 3C

Figure 3c: Gluma CPS Primer (Surface and Fractured views)

FIG 3D

Figure 3d: Solobond Plus (Surface and Fractured views)

FIG 3E

Figure 3e: All Bond 2 (Surface and Fractured views)

FIG 3F

Figure 3f: Scotchbond (Surface and Fractured views)

Application of a Hema-Seal G primer resulted in limited deposition on the dentine disc with small deposits partially occluding the tubule orifices (Figure 3a). After comparison with the control half of the disc, it was observed that there was some apparent further etching. When the dentine disc was fractured the tubule orifices appeared unsealed with no sign of material deposit although some tubules appeared widened (arrow). The Gluma 3 primer produced partial occlusion of some of the tubule orifices with irregularly shaped deposits, usually observed on the surface and at the edges of the tubule openings (Figure 3b arrow). When fractured, tubule lumens free of deposits were observed. After treatment with Gluma CPS there were sparse irregular deposits on the dentine surface, some of which appeared to partially occlude the tubule orifices, whereas others were observed on the tubule periphery (Figure 3c). Fracture of the disc revealed little or no tubule occlusion or penetration.

Solobond Plus Universal Adhesive System was applied on a dentine disc etched with Vococid. The dentine surface appeared rough, and the tubule orifices were patent (Figure 3d). Some particles were noticed on the dentine around, but not occluding, the tubules (arrow). When fractured the tubules, small deposits 20-30µm were observed deeper into the tubules, although the tubules were open and widened towards the dentine surface fractured compared to the control disk (example 2a).

After application of All Bond 2 to the dentine it was apparent that the surface was covered with an almost uniform layer sealing the tubule orifices (Figure 3e). Fracture of the dentine disc revealed the presence of a thin layer covering the tubule orifices which appeared to coat the inner walls of the dentine tubule to a varying depth (arrow). Treatment with Scotchbond following etching with maleic acid 10% for 15 seconds demonstrated that some of the ttubule orifices were evident, but greatly reduced in diameter (Figure 3f). The fractured view revealed a degree of product penetration within the tubule lumen which appeared coated (Figure 3f fractured view). One Step produced an even layer which covered the whole dentine surface and no tubule orifices were obvious (Figure 3g). Fracturing the dentine disc showed a thin layer covering the tubule orifices and a considerable degree of agent penetration within the tubule lumen (arrow).

FIG 3G

Figure 3g: One Step (Surface and Fractured views)

Subjective assessment of the test products’ ability to both occlude and penetrate the dentinal tubules was recorded as shown in Table 2. These results would suggest that the fluoride varnishes (Bifluoride 12 and Duraphat, and Cervitec) were effective in both occluding and penetrating the dentine tubules. It was also evident that All Bond 2, One Step and Scotchbond primers were superior to those of the HEMA group primers (Hema-Seal G, Gluma 3, Gluma CPS) and Solobond Plus

Table 2: Summary of the test products’ tubule occluding and penetrating ability

Test agents

Degree of tubule occlusion

Degree of dentine surface coverage

Degree of tubule lumen penetration

Bifluoride 12

+++

+++

+++

Cervitec

+++

+++

+++

Duraphat

+++

+++

++

Hema-Seal G

+

+

0

Gluma 3 Primer

+

+

0

Gluma CPS

+

++

0

Solobond Plus

+

+

0

All Bond 2

+++

+++

+++

Scotchbond

++

++

+++

One Step

+++

+++

+++

Key:
+++ Most tubules occluded/maximum surface coverage/good penetration
++ Some tubule occlusion/some surface coverage/some penetration
+ Few tubules occluded/little surface coverage/little penetration
0 No occlusion/coverage/penetration.

Discussion

Currently, the most accepted mechanism of stimulus transmission across dentine is the hydrodynamic theory, which proposes that rapid shifts of fluid movement in either direction within the dentine tubules may stimulate mechano-receptors in or near the pulp to excite the pulpal nerve and cause pain. This theory leads to the concept of dentine tubule occlusion as a method of dentine desensitization [2,3].

The use of the dentine disc model has been proved to be a reliable method for the initial in vitro screening of tubule occluding properties of potential desensitizing agents [5,7,8]. The dentine disc would appear to be the method of choice since it is easy to use, reproducible, provides a flat surface for elemental analysis and may be correlated with fluid flow research [7]. Due to the differences in the size, orientation, density and diameter of the dentine tubules throughout the tooth [9], the discs were obtained from the same region of the tooth, below the crown and above the root canal. One half of each dentine disc provided the control and the other half the test portion, and only those tubules from the central region of the disc, on either side of the fractured edge, were examined and all the observations on the test side were compared with the control half of the same disc. All the tested agents were applied according to the manufacturer’s instruction in an attempt to mimic the clinical situation.

Cervitec is a chlorhexidine containing varnish, marketed in Europe, which possesses very good dentine covering and tubule occluding properties. Chlorhexidine may act as an antiseptic in the dentine tubule lumen and reduce the number of microbes penetrating the open tubules of sensitive dentine, reducing possible pulp inflammation.

Duraphat has been studied in vivo [10-18] and proved to be effective in alleviating DS short-term. The main ingredient of Duraphat is sodium fluoride (NaF), which can precipitate onto the dentine surface [19] and may contribute its tubule occluding potential, along with its fluid texture which allows penetration into the tubule lumen. Fluoride is also reported to have potential for reducing sensitivity, perhaps acting on the SNA, and the effect of the combined sensory and physical actions may account for the widespread clinical success of Duraphat.

Bifluoride12 appears to seal the dentine surface with a crystal-like deposit in an amorphous matrix that penetrates the tubule lumen. It contains sodium and calcium fluoride, which probably account for the presence of deposits. Although the plugs penetrating the tubules do not completely occupy the tubule lumen, they appear to reduce the tubule radius and this, in association with the desensitising potential of the fluoride, may contribute to a reduction in DS in the clinical environment.

The Gluma group primers (Gluma 3, Gluma CPS), Hema-Seal G and Solobond Plus primer contain mainly glutaraldehyde and HEMA. They appeared to produce similar deposits on the dentine disc surface, partially occluding the tubule orifices. Fracturing did not reveal any degree of deposit penetration. Dondi Dall’ Orologio et al. [20] reported significant reduction of DS following application of Gluma 3 attributed to a possible reaction of glutaraldehyde with the dentinal fluid proteins, precipitation and thus a partial or complete obturation of dentine tubules. The mode of action of HEMA is still unknown. Probably, the partial occlusion of tubule openings shown in the present study may explain the reduction in DS. Hema-Seal G also contains sodium fluoride. Thus any reduction in DS may be attributed either to the partial occlusion of dentine tubule orifices or to the action of sodium fluoride which forms crystals of calcium fluoride reducing the radius of dentine tubules [19]. However, a degree of ‘over etching’ was observed following application of Hema-Seal G, indicating that, when applied to the dentine surface in vivo and in conjunction with acidic dietary intake, it might result in an increase in dentine permeability.

All-Bond 2 primers formed a layer that appeared to fully cover the dentine surface and also penetrated the tubule lumen to some depth. This is in accordance with Tay et al. [21] who also showed penetration of the tubule lumen. Furthermore, Ianzano et al. [22] and Gillam et al. [23] reported that All-Bond 2 primers were effective in vivo in reducing DS, although different evaluation techniques were employed by these investigators. All-Bond 2, therefore, may be useful in the in-office treatment of DS although its effectiveness may be relatively short-lived. One Step was a relatively new product (at the time of evaluation) and so far there are no studies evaluating its desensitizing or occluding properties although it is the same as All Bond 2. As with All-Bond 2, One Step may be of some clinical value given that it’s occluding and penetrating properties are satisfactory and it is also easily applied. Scotchbond has been reported to be effective in reducing DS [24] although it is possible that the primer was subsequently covered with a light-cured resin. In the present study only, the primer was applied onto the dentine disc surface and the results showed a degree of surface coverage although some tubules with reduced diameters were also evident. This may be due to the application procedure or the etching period. It is possible that longer application period or thicker layer would completely obscure the tubule openings. However, the fracture of the disc revealed penetration of the product within the tubules rendering this agent a potential desensitiser.

This study was based on a well-established dentine disc model which has been modified to provide greater control in the preparation and analysis of surface deposits. In this respect, the method may be considered an improvement over previously reported studies. Difficulties may still arise, however, in the interpretation of the results particularly with regard to fracturing the disc halves, which is relatively technique sensitive, and the exact plane of fracture is somewhat unpredictable. Nevertheless, if the control and the test disc sections are carefully compared, the penetrating potential of the tested agents can be satisfactory evaluated as indicated in this present study. The dentine disc model although useful for the initial in vitro assessment of different agents may not reflect the in vivo situation and the results of this in vitro study should be extrapolated to the in vivo situation with caution. Factors such as pulpal pressure, the oral environment (saliva, gingival fluid) and patient’s habits (vigorous toothbrushing, acidic dietary intake) may also influence the retention of these products on the dentine surface. The presence of dentinal fluid within the tubules and its rate of movement in vivo may influence the formation of deposits on the dentine surface despite efforts to keep the exposed dentine [15]. Vongsavan and Matthews [25] reported that the penetration of different molecules into the dentine tubules may be greater in vivo than in vitro.

All the agents introduced in the present study, appear to be clinically applicable since they are easily applied, and some appear to possess both occluding and penetrating ability. Varnishes and primers, (together with sealants which were not reported on in this study), because of the relative ease of application, may provide a useful treatment option for the practicing dentist in the alleviation of DS. Attention should be paid to the mode of their application, which may be technique sensitive, e.g. All Bond 2 and Scotchbond where a vigorous air blast may remove a great amount of the agent leaving uncovered dentine tubules. Tubule penetration properties are important because even when the agent is removed from the dentine surface, plugs of the material may remain within the tubule lumen, which may either maintain the desensitizing effect of the agent or provide the patient with temporary relief until natural tubule occlusion occurs. Both short and long term clinical studies are required to determine whether the in vitro potential of these agents can be extrapolated in the clinical environment.

The majority of in vitro studies have evaluated the desensitizing properties of agents using descriptive methods of dentine surface assessment. Although some authors claimed that their methods of assessing the amount of tubule occlusion were quantitative in nature [6,26] in many cases they were semi-quantitative with no measuring involved. Quantitative studies using digital image analysis and SEM would provide a more accurate result [27] however, most of the products in this study resulted in total surface coverage, negating the need for measurement. Studies including fractures of dentine discs are of benefit in the investigation of the desensitizing potential as there may be blockage within the tubule which is not evident from the surface. Elemental analysis was not within the scope of this study, but further examination of some of the surface products would be interesting.

Conclusions

The results of this study suggest that, of the agents tested, the varnishes, All-Bond 2 primer, Scotchbond primer, and One Step appear to have both occluding and penetrating properties. However, agents which have been observed to be effective in vivo but with no significant tubule occluding or penetrating properties as demonstrated in the present study may employ other mechanism/s which cannot be simulated in this in vitro model. Moreover, the results obtained following the application of all the agents tested in the present study may be affected by the application technique(s) employed to apply them on the dentine surface, although every effort was made to follow the manufacturer’s instructions for clinical application.

References

  1. Addy M, Mostafa P, Absi EG, Adams D (1985) Cervical dentine hypersensitivity. Etiology and management with particular reference to dentifrices. In: Rowe, N. H. (ed): Proceedings of symposium on hypersensitive dentine Origin and Management, University of Michigan, (ed) E. & S, Livingston Limited, Edinburgh & London, pp: 147-167. [crossref]
  2. Gillam DG (1995) Mechanisms of stimulus transmission across dentin–a review. J West Soc Periodontol Periodontal Abstr 43(2): 53-65. [crossref]
  3. Brannstrom M (1962) The elicitation of pain in human dentine and pulp by chemical stimuli. Arch Oral Biol 7: 59-62. [crossref]
  4. Pashley DH (1985) Strategies for clinical evaluation of drugs and/or devices for the alleviation of hypersensitive dentin. In: Rowe NH, editor. Proceedings of Symposium on Hypersensitive Dentin. Origin and Management. Ann Arbor: Univ Michigan. p.65-88 [crossref]
  5. Greenhill JD, Pashley DH (1981) The effects of desensitizing agents on the hydraulic conductance of human dentin in vitro. J Dent Res 60: 686-98 [crossref]
  6. Absi EG, Addy M, Adams D (1995) Dentine hypersensitivity: uptake of toothpastes onto dentine and effects of brushing, washing and dietary acid–SEM in vitro study. J Oral Rehabil 22(3): 175-82. [crossref]
  7. Mordan NJ, Barber PM, Gillam DG (1997) The dentine disc. A review of its applicability as a model for the in vitro testing of dentine hypersensitivity J Oral Rehabil 24(2): 148-156. [crossref]
  8. Ling TY, Gillam DG, Barber PM, Mordan NJ, Critchell J (1997) An investigation of potential desensitizing agents in the dentine disc model: a scanning electron microscopy study. J Oral Rehabil 24(3): 191-203. [crossref]
  9. Pashley DH, Leibach JG, Horner JA (1987) The effects of burnishing NaF/kaolin/glycerin paste on dentin permeability. J Periodontol 58(1): 19-23. [crossref]
  10. Kerns DG, Scheidt MJ, Pashley DH, Horner JA, Strong SL, Van Dyke TE (1991) Dentinal tubule occlusion and root hypersensitivity. J Periodontol 62(7): 421-8. [crossref]
  11. Kim S (1986) Hypersensitive teeth: desensitization of pulpal sensory nerves. J Endod 12(10): 482-5. [crossref]
  12. Sena FJ (1990) Dentinal permeability in assessing therapeutic agents. Dent Clin North Am 34(3): 475-90. [crossref]
  13. Knight NN, Lie T, Clark SM, Adams DF (1993) Hypersensitive dentin: testing of procedures for mechanical and chemical obliteration of dentinal tubuli. J Periodontol 64(5): 366-73. [crossref]
  14. Jain P, Vargas MA, Denehy GE, Boyer DB (1997) Dentin desensitizing agents: SEM and X-ray microanalysis assessment. Am J Dent 10(1): 21-6. [crossref]
  15. Dragolich WE, Pashley DH, Brennan WA, O’Neal RB, Horner JA, Van Dyke TE (1993) An in vitro study of dentinal tubule occlusion by ferric oxalate. J Periodontol 64(11): 1045-51. [crossref]
  16. Gwinnett AJ (1988) Aluminium oxalate for dentin bonding. A SEM study. Amer J Dent 1, 5-8. [crossref]
  17. Ellingsen JE, Rölla G (1987) Treatment of dentin with stannous fluoride – SEM and electron microprobe study. Scand J Dent Res 15, 281-286. [crossref]
  18. Hansen EK (1992) Dentin hypersensitivity treated with a fluoride-containing varnish or a light-cured glass-ionomer liner. Scand J Dent Res 100(6): 305-9. [crossref]
  19. Addy M, Mostafa P (1989) Dentine hypersensitivity. II. Effects produced by the uptake in vitro of toothpastes onto dentine. J Oral Rehabil 16(1): 35-48. [crossref]
  20. Dondi dall’Orologio G, Malferrari S (1993) Desensitizing effects of Gluma and Gluma 2000 on hypersensitive dentin. Am J Dent 6(6): 283-6. [crossref]
  21. Tay FR, Gwinnett AJ, Pang KM, Wei SH (1994) Structural evidence of a sealed tissue interface with a total-etch wet-bonding technique in vivo. J Dent Res 73(3): 629-36. [crossref]
  22. Ianzano JA, Gwinnett AJ, Westbay G (1993) Polymeric sealing of dentinal tubules to control sensitivity: preliminary observations. Periodontal Clin Investig 15, 13-16. [crossref]
  23. Gillam DG, Coventry JF, Manning RH, Newman HN, Bulman JS (1997) Comparison of two desensitizing agents for the treatment of cervical dentine sensitivity. Endod Dent Traumatol 13: 36-39. [crossref]
  24. Copeland JS (1985) Simplified remedy for root sensitivity. Northwest Dent 64(6): 13-4. [Crossref]
  25. Vongsavan N, Matthews B (1991) The permeability of cat dentine in vivo and in vitro. Arch Oral Biol 36(9): 641-646. [crossref]
  26. McAndrew R, Kourkouta S (1995) Effects of toothbrushing prior and/or subsequent to dietary acid application on smear layer formation and the patency of dentinal tubules: an SEM study. J Periodontol 66(6): 443-8. [crossref]
  27. Ahmed TR, Mordan NJ, Gilthorpe MS, Gillam DG (2005) In vitro quantification of changes in human dentine tubule parameters using SEM and digital analysis. J Oral Rehabil 32(8): 589-97. [crossref]

Faking Orgasm: Interviews with College Women about How, When, and Why They Pretend to Experience Orgasm

DOI: 10.31038/AWHC.2023623

Abstract

This study investigated the context in which young women fake orgasm, as well as their reasons. A key component of this study was to examine the way that sexual scripts and societal pressures influence young women who fake orgasm. In-depth interviews were conducted with 12 college women between the ages of 19-22 who had faked orgasms during sexual intercourse. Interview transcripts were analyzed for common themes. These college women faked orgasm for a variety of reasons and in the context of a diversity of relationships. Limitations, suggestions for future research, and recommendations for sexuality education are discussed.

Introduction

Faking orgasm, also known as feigning or pretending to orgasm, has been seen in popular media for decades. When Harry met Sally, was the first movie to highlight a woman faking orgasm. In the iconic restaurant scene, Sally claims that “at one time or another most women have faked it” [1]. Television shows, including Seinfeld [2], Parks and Recreation [3], and I Am Cait [4] have all dealt with the topic, and popular magazines, such as Cosmopolitan, have feature articles on faking orgasm including “7 Sad But True Reasons Women Fake Orgasm” [5] and “Funny Ways People Faked” [6]. These examples from popular media acknowledge that faking orgasm is something some women do, and at the same time raise important questions for researchers about how common it truly is and the reasons why women fake orgasm. It has been suggested it is one of the “least well-studied human sexual behaviors” [7].

Prevalence of Faking Orgasm

Several researchers have looked at the prevalence of women faking orgasm (e.g., Caron, 2021; Darling & Davidson, 1986; Ellison, 2000; Fahs, 2014; Hite, 1976; Muehlenhard & Shippee, 2010; Wiederman, 1997) [8-14]. Four of the earlier studies to address the prevalence of women faking orgasm provided important groundwork for more recent studies. One of these earlier studies was the research by Shere Hite published in her 1976 book, The Hite report: A nationwide study of female sexuality, which was based on the responses of 3,019 women, ages 14 to 78 [12]. The Hite Report revealed that most women experienced orgasm during clitoral stimulation rather than through vaginal penetration (a somewhat revolutionary finding for that time) and that these women felt that men’s attitudes about sex needed to expand to include the stimulation women needed. Important to the current study was her finding that more than half (57%) of women reported faking orgasm [12]. A decade later, a second study was published by Darling and Davidson specifically focused on faking orgasm (referred to as “pretending orgasm”). Their sample included 868 nurses from around the United States and revealed that, of the 745 women who had engaged in penile-vaginal intercourse, more than half (58%) had pretended to orgasm [9]. Another decade later, a third study focusing on faking orgasm involved a sample of 161 college women ages 18-27 from a Midwestern state university [14]. His findings were consistent with the two previous studies in that more than half (56%) of the women reported faking an orgasm during sexual intercourse [14]. In 2000, a fourth study discussing women faking orgasm was published by Ellison in her book, Women’s sexualities: Generations of women share intimate secrets of sexual self-acceptance. Similar to Hite, Ellison reported the results of her national survey, as well as in-depth interviews with women [10]. Of the 2,311 women between the ages of 23 and 95 who answered the questions about faking orgasm, 70% indicated they had faked an orgasm at least once with their partner [10].

More recent studies in the last two decades confirm what these earlier studies have shown: many women have faked orgasm. Muehlenhard and Shippee’s 2010 study of Kansas University college women from the ages of 18-29 found that two-thirds (67%) had faked during intercourse. Their study also looked beyond sexual intercourse and found that some college women pretended to orgasm during oral sex, manual stimulation, and phone sex [13]. Fahs’ (2014) research involving in-depth interviews with 20 women ages 18-59 found that 75% of women had faked orgasm at least once in their life [11]. Nearly half (45%) of these women reported that they faked orgasm regularly during their sexual encounters [11]. And in 2021, Caron published her book, The sex lives of college students: Three decades of attitudes and behaviors based on her annual survey of college students at the University of Maine. Her findings for the 6,654 students ages 18-22 covered a wide range of sexuality topics. In terms of faking orgasm, 70% of sexually active college women in the study said they have faked an orgasm. This is the only study to look at faking orgasm over time. When looking across the 30 years, Caron found that for college women, faking behavior has increased dramatically over time – from less than half to over three-quarters of college women today saying they have faked an orgasm [8].

Reasons Why Women Fake Orgasm

Along with research on the prevalence of women faking orgasms, there are several of studies that have examined the reasons why women fake orgasm. (i.e., Bryan, 2001; Braun et al., 2014; Fahs, 2014; Kaighobadi et al., 2010; Lafrance et al, 2017; Mialon, 2012; Muehlenhard & Shippee, 2010; Roberts et al., 1995) [11,13,15-20]. In addition, Cooper et al. (2014) developed a scale to assess women’s motives for faking orgasm, The Faking Orgasm Scale for Women [7]. These reasons have ranged from concern for their male partner’s feelings and wanting sex to end, to saving the relationship and pressure to perform in order for her to appear “normal.”

The most common reason researchers have found for why a woman might fake orgasm has been out of concern for the feelings of her male sexual partner; women reported they faked orgasm in order to preserve their partner’s feelings, or in some cases, to give him an “ego boost.” This was labeled by Cooper et al. (2014) as “altruistic deceit” and based on gendered beliefs, such as if a woman does not orgasm it will negatively impact the man’s ego. In Roberts et al.’s 1995 study, some of the 75 women interviewed explained how their partner felt he had failed in some way if she did not orgasm, so faking was a way to ensure he did not feel like a failure [20]. Ellison (2000) also found that women viewed faking orgasm as important in taking care of their partner’s feelings. One of the women interviewed stated, “he had to think he was a good lover, satisfying me. Basically, faking was me taking care of the man” [10]. In Bryan’s 2001 study, the women that she surveyed described wanting to boost their partners’ egos and not hurt his feelings. Her orgasm indicated whether or not he was sexually proficient and skilled; it emphasized that sex is a skilled activity, and a woman’s orgasm was the man’s responsibility [15]. Muehlenhard and Shippee (2010) found that 78% of women had pretended orgasm in order to avoid hurting their partner’s feelings, while 47% of women faked orgasm to make their partner feel good about themselves [13]. Mialon (2012) also found that women who care about their partner’s sexual pleasure are more likely to fake; they want to make him happy [19]. And along with protecting his feelings, a woman may want to reinforce her partner’s sexual skills. Fahs found that these were common themes in both her 2011 and 2014 research [11,21]. In Braun et al.’s 2014 research, a common reason women faked orgasm was to save the feelings of their most intimate partner [16]. Most recently, a study of 462 women ages 19 to 73 in the UK found that those who held more gendered beliefs, such as women’s orgasm is necessary for men’s gratification, were more likely to fake orgasm [22].

A second common reason some women fake orgasm is because she wants sex to end (Cooper et al. (2014) labeled this “sexual adjournment” [7]. As a woman in Ellison’s study (2000) described, “I enjoy sex even without orgasm. I fake because my partner tries too hard to get me to orgasm and that turns me off” [10]. In Bryan’s 2001 study, many women faked orgasm in order to stop intercourse [15]. Their reasons for why they wanted to end intercourse included feeling tired, bored, nervous, or in pain [15]. Muehlenhard and Shippee (2010) found that 61% of women in their sample indicated that wanting sex to end was their reason for faking orgasm. The women who wanted sex to end claimed that it was because “they were bored, not in the mood, or tired and wanting sleep” (Muehlenhard & Shippee, 2010). These researchers also found that some women faked orgasm so that their partner would orgasm, and this would lead to the end of sex [13]. In both Fahs’ 2014 study and Lafrance et al.’s 2017 study, women said they purposely faked orgasms in order to end sexual encounters [11,18].

A third common reason found in studies of why women fake an orgasm included her feeling that this keeps her partner happy and the relationship intact. Some women described feeling as though they could lose their boyfriend if they don’t fake [20]. In Ellison’s 2000 study, she describes this as a common theme. One interviewee explained the reason she fakes orgasm is because she needs to please her partner so that ‘then he won’t leave me and I’ll have a boyfriend” [10]. In Bryan’s 2001 study, the researcher found that there were a broad variety of partner-related reasons, and these included, “to maintain the relationship” [15]. In Kaighobadi et al.’s 2010 study about women faking orgasm in order to retain their mate, they found that women who were more insecure in their relationships and who perceived a higher risk of partner infidelity were more likely to fake orgasm [17].

Finally, some researchers have found that the reason a woman has faked orgasm was to convince their partner that she is normal; otherwise the feeling is that there may be something wrong with her. Bryan (2001) found that some women were motivated to fake orgasm because they felt embarrassed, ashamed, or abnormal because they were not going to have an orgasm when they thought they should [15]. Bryan’s research found that women “feared appearing inexperienced or naieve;” therefore, they felt pressure to experience an orgasm [15]. Along with feelings of abnormality come feelings of guilt or shame. In Fahs’ 2014 research, some women felt her inability to orgasm implied that there was something wrong with her, as one woman explained, “I don’t want him to know that I’m one of those women who can’t get aroused from a penis inside of her” [11].

Sexual Scripts

As these studies reveal, faking orgasm is not only common, but increasing, and women of all ages indicate a variety of reasons for faking, including concern for a partner’s feelings, wanting sex to end, to save the relationship, and pressure to appear “normal.” However, the idea that today’s young women fake orgasms seems nonsensical when viewed through a modern lens.

In a society that has seen a rise in feminism, discussions of women’s empowerment and sexual pleasure, women’s marches, empowering television shows and movies, and a myriad of information available online about sexual functioning and women’s pleasure, one would think that the number of women faking orgasm would be small or have decreased over the past few decades, not increased [23]. On the other hand, sex seems to be everywhere, but is seldom talked about honestly and openly. For example, pornography is accessible, but too often unrealistic, and sex education rarely includes discussions about female pleasure [21,24]. It is no wonder that, according to Casey et al. (2013), traditional sexual scripts for women are still in existence. These scripts include women not desiring sex, having a weak “sex drive,” resisting advances, being more highly valued if she is less sexually experienced, as well as the notion that women should prefer relational sex, want commitment and monogamy, and should seek emotional intimacy and trust with sex [25].

These traditional sexual scripts also provide ideas surrounding women’s experience with orgasm. According to Lavie-Ajayi and Joffe (2009) [26] in their article Social Representations of Female Orgasm, these scripts center on three ideas: 1) women’s orgasm as the central indicator of sexual pleasure and the goal of sex, 2) vaginal orgasms are thought of as better than clitoral orgasms, and 3) while it is common for women not to experience orgasm through intercourse, if she doesn’t, it is a sign of her deficiency. These orgasm scripts suggest reasons why women might fake during a sexual encounter. Essentially, the message for women has been that “we have been taught that orgasms are obligatory” [27]. Women, as well as men, have been socialized to believe that orgasm is the goal of sex, and without an orgasm, sex is pointless. This puts an incredible amount of pressure on both parties involved, and it may be easier to fake an orgasm than to deviate from these social norms [26].

The Purpose of this Study

While a number of studies have been done over the past several decades on the frequency of and reasons for faking orgasm, many involve women across a range of ages. Few studies have examined younger women. This qualitative study investigated the context in which today’s young women fake orgasm and examined their reasons for faking orgasm. A key component of this study was to explore the ways that traditional sexual scripts and societal pressures may continue to influence college women who fake orgasm. Specifically, this study involved in-depth interviews with younger college women who have faked orgasm during sexual intercourse. The following research questions were explored:

  1. How often and in what context do college women fake orgasm?
  2. What are college women’s reasons for faking orgasm?
  3. How do college women feel about faking and what is their sense of how their partner(s) and friends feel(s) about women faking orgasm?
  4. What do the college women’s accounts of faking orgasm reveal about their sexual scripts?

Methodology

The Sample

The criteria for inclusion in the study were college women ages 18-22 who had faked an orgasm during sexual intercourse. All 12 women described faking orgasm in the context of a sexual relationship with a man. Participants were recruited through announcements made in classes and e-mail announcements sent to several university discussion boards. The final sample included 12 women (Table 1) attending a public university in the northeastern United States. The age of the participants ranged from 19 to 22 years old, with the average being 21 years old. All 12 of the participants identified as White and cisgender. Ten of the participants identified as heterosexual, one identified as bisexual, and one identified as queer. The number of sexual partners each participant had ranged from 1 to 35 (M=11.6; SD=11). The number of sexual partners each participant faked with ranged from 1-20 (M=6.4; SD=6.8). Most women reported faking orgasm with most of their sexual partners. The percentage of partners they faked orgasm with ranged from 14% to 100% (M=65.4%; SD=26.3).

Table 1: Demographics of Participants (N=12)

Participant Pseudonyms

Age

Race

Sexual Orientation

Number of Sexual Partners*

Number of partners they faked with

Percent of Partners Faked With

Abigail

22

White

Straight

1

1

100%

Bella

21

White

Straight

6

4

66%

Charlotte

21

White

Straight

7

7

100%

Delilah

21

White

Straight

1

1

100%

Emma

22

White

Straight

27

20

74%

Faith

21

White

Straight

3

2

66%

Grace

21

White

Straight

5

3

60%

Hannah

22

White

Bisexual

9

2

22%

Irene

19

White

Straight

3

2

66%

Jasmine

22

White

Straight

22

3

14%

Kaylee

19

White

Queer

20

12

60%

Lily

22

White

Straight

35

20

57%

*All reported that their sexual partners were male

Interview Procedure

The interview was comprised of a series of questions focusing on the experience of faking orgasm. The Human Subjects Committee of the university approved the interview protocol. Interviews began with basic demographic questions (e.g., age, race, number of sex partners). The subjects were then asked a series of questions that centered on how often and the context for faking, what their reasons were for faking, how they and their sexual partner(s) and friends felt about women faking orgasm, and their sexual scripts.

College women who met the criteria (age 18-22 who had faked an orgasm during sexual intercourse) and expressed interest in participating in the study were provided with a copy of the consent form before an interview was scheduled. Consent was implied when they agreed to arrange an interview after reviewing the consent form. Consent was also verbally provided by the participants at the start of the interview. Interviews were conducted face-to-face in a private space on the university campus. Participants were assured that any and all responses would be kept confidential. No names or identifying information was included in the data report; a pseudonym was assigned to each participant, and any names of partners or friends mentioned in the interview were changed. Each interview lasted approximately ninety minutes. Interviews were tape recorded and extensive notes were taken. At the end of the interview, participants were given the opportunity to review the notes with their responses for omissions or clarifications. The taped interviews were later transcribed and afterwards the content from the taped interview was deleted.

Interview Analysis

Interview transcripts were analyzed using established methods of qualitative inquiry, including coding and categorizing processes that make use of both deductive and inductive approaches [28]. Thematic analysis was utilized to analyze and code the interviews. The first and second author read the first three interview transcripts independently, and initial coding of the transcripts involved marking comments that fit under the four research questions of focus for the study: frequency/context, reasons, feelings, and scripts. For example, a response that encompassed a woman’s reason for faking orgasm would have the initial code word “reason” added next to it. Next, coding categories or themes were created under these areas of focus through a process of open coding [28]. For something to be considered a theme, at least half of the women (six women) needed to discuss this in their responses. For instance, under the responses for “reasons,” if six or more women mentioned that the reason she faked an orgasm was because she worried about her partner’s ego and/or did not want to hurt his feelings, this was categorized under that theme. These were compared and coding categories were further developed.

The rest of the interviews were coded by the first author and reviewed by the second author as any changes or additions were made. No new coding categories emerged after approximately 10 interviews were completed, leading to assurances of content saturation. A total of 16 themes emerged from the coding. These are discussed below and presented in Table 2. In terms of frequency and context for faking orgasm, four themes were identified. For reasons for faking orgasm, five themes were determined, and for feelings about faking orgasm a total of four themes were identified. Finally, the fourth research question sought to understand what young women’s accounts of faking orgasm reveal about their sexual scripts. Three sexual scripts were identified and will be discussed.

Table 2: Themes identified for college women’s frequency and context of faking orgasm, reasons for faking orgasm, feelings about faking orgasm, and sexual scripts related to faking.

Frequency and Context of Faking Orgasm

·         How often: It depends

·         How she faked: I was a great actress and lied

·         When she faked: It was typically with intercourse

·         Who she faked with: When I didn’t know him well

Reasons for Faking Orgasm

·         I did not want to hurt his feelings

·         I was uncomfortable being with him

·         I felt I was taking too long

·         He did not know what he was doing

·         It’s expected

Feelings about Faking: Hers, Her Partner(s), and Her Friends

·         I feel it is pretty normal

·         I feel guilty, but it was necessary

·         My sexual partner would feel deceived

·         My girlfriends feel it is perfectly okay

Sexual Scripts

•         Media messages of what “normal sex” looks like

•         Her orgasm is a reflection of his competence

•         Her orgasm signals that sex is complete

Findings

Themes are reported below for each of the areas of focus for this research: frequency and context of faking orgasm, reasons for faking orgasm, feelings about faking orgasm, and sexual scripts. Direct quotations from the interview transcriptions are included to highlight the findings (Table 2).

Frequency and Context of Faking Orgasm

The first research question asked, “How often and in what context do college women fake orgasm?” The 12 college women were asked a series of questions to understand their experience of faking orgasm. Four unique themes were identified from the in-depth interviews, including how often she faked, how she faked, when she faked, and who she faked orgasm with. These are described below and are listed in Table 2.

How often: It depends. The first theme related to the frequency and context of faking orgasm was related to how often the women faked an orgasm. When asked how often they had faked an orgasm, the most common response was “it depends,” with responses ranging from “only a few times” to “hundreds of times.” For many women, it depended on a range of factors from how inexperienced she was or how inexperienced her partner was, to how well she knew him or how much pressure she felt. While some women said they had faked an orgasm only a few times in one relationship, others indicated they had faked “hundreds of times” in most relationships. Examples of how college women described how often they faked an orgasm are included below:

I did it more at the beginning of our relationship, and then less later into the relationship because I think that we both started learning more what we liked in the bedroom. I would say almost the majority of the time at the beginning of the relationship and then almost not at all as the relationship progressed forward. (Grace)

I would say that every one-night stand that I have had, I pretty much faked with. Like when I have a one-night stand, I would say it’s a guarantee pretty much that I am going to fake. It’s usually different than when I’m with someone that I’m emotionally close to. (Lily)

How she faked: I was a great actress and lied. The second theme related to frequency and context of faking orgasm focused on how she actually faked her orgasm. The majority of women described how faking orgasm involved good acting and included descriptions of the kinds of actions (i.e., body movements and sounds) they would engage in. They also said that when asked by their partner after sex if they had had an orgasm, they deliberately lied to him. Many not only described being good at faking, but also discussed getting better overtime with more sexual experiences. Some sample quotes describing how she faked include:

I verbalize, like moaning and getting grabbier. Definitely working myself up kind of thing and then heavy breathing afterwards. That was always a definite and I was like “Oh my God” ((panting)) kind of thing. I would say that I usually didn’t say anything though, just used my muscles and I would scratch their back too. (Kaylee)

I didn’t use just my body. He was like, ‘Did you get it?’ and I just said “Yes” even though I didn’t, so most of the time it’ll be like physical and moaning except for the two times that I straight up lied out of my mouth. (Charlotte) When she faked: It was typically with intercourse. A third theme related to frequency and context of faking focused on sexual intercourse. All 12 college women described faking orgasm during penile-vaginal intercourse. Two women added that they had also faked an orgasm during oral sex. Some women described that they began faking orgasms during sexual intercourse before they had ever had a real orgasm, but once they experienced an orgasm and knew what it felt like, they got better at faking. Some examples of women explaining when they faked include:

I fake during intercourse because I don’t even have a reason. Oral depends, I don’t know, I fake during both of them, but most commonly during intercourse because like I said, I don’t orgasm just by intercourse alone, so, uh, it is to make him feel better. (Delilah)

I emphasized more or got more convincing when I was faking when we were having sex. I think that because I knew what the orgasm was supposed to feel like because of a previous relationship, I was better at faking when I was doing stuff with other guys. (Irene)

Who she faked with: When I didn’t know him well. The fourth and final theme related to frequency and context of faking centered on how well they knew their sexual partner. All 12 college women talked about how they faked orgasm with someone they did not know well, either in the context of a one-night stand or in the very beginning of the dating relationship. It was described by many as “easier to get away with” if they faked an orgasm with someone whom they did not know and would never see again. Most described faking at the start of a relationship that eventually evolved into something long-term. As their partner got to know their sexual needs, there was no need to fake anymore. Some examples describing who women faked with include:

We were dating for less than a year, I only did it like a couple of times in the beginning ‘cause I kind of almost felt bad ‘cause it’s like obviously you’re putting a lot of effort in, you really are trying. It was just sometimes it was not going to happen and also that was one of my early relationships so I wasn’t as comfortable talking openly about it so I didn’t feel okay asking him to try something different. (Jasmine)

I only dated him for like three months, it was a stupid relationship. We didn’t have sex very many times, so this is kind of awkward, but I faked every single time with him. We only had sex like three times. This is so sad. I never even had an orgasm with him. (Bella)

Reasons for Faking Orgasm

The second research question asked, “What are college women’s reasons for faking orgasm?” A series of interview questions explored their most important reasons and what their rationale was for faking orgasm. Five themes were identified for the reasons women said they fake orgasm, including not wanting to hurt his ego, experiencing uncomfortable emotional and physical feelings with him, worrying she was taking too long, feeling like the guy wasn’t doing anything that was stimulating her, or she felt it was expected that she reach orgasm as part of sex. Each woman had many reasons for faking (not just one) and their reasons varied based on the context of the sexual experience. These five themes are described below and listed in Table 2.

I did not want to hurt his feelings. The first theme related to reasons for faking included concern for his feelings. Most of the college women described faking orgasm because they were worried about his ego and whether or not he would feel like he was a good sexual performer. Examples of things women said included:

I guess mainly I was just trying to make my partners feel better about themselves and that’s why I would never say, “Hey I’ve been faking the entire time we’ve been sleeping together” because that would take away all that validation too. Plus, I feel like it’s kind of a waste of time to be faking quite a bit and then tell the partner that you were faking. Their feelings are really going to get hurt that way. (Hannah)

I think that men get their feelings hurt easily and they’re very sensitive when it comes to sex and they always want to protect their ego and I think that there’s at least in some way pressure on women to fake orgasms so that they protect the feeling of their male partner. (Abigail)

I was uncomfortable being with him. The second theme related to reasons for faking orgasm was because they were really uncomfortable being with their sexual partner and therefore knew the sexual encounter was not going to lead to orgasm. They described being emotionally uncomfortable in terms of not being able to relax with him, not feeling a connection to him, or feeling very self-conscious. Many women also mentioned feeling physically uncomfortable (i.e., pain) and faking orgasm in order for the sex to end. For example:

I think “discomfort” would be the one-word summary of why I faked. I think with the more casual relationships, I never was emotionally close to them so I was emotionally uncomfortable with the situation. I felt like, “I don’t like that you’re seeing my boobs right now and I hardly know you.” With my first boyfriend it was probably more like, “This is my first partner ever, I don’t know what I’m doing.” I’m self-conscious, that’s why I’m uncomfortable and also when you don’t really know what you’re doing. I think physically that’s uncomfortable too. (Charlotte)

I felt like I was taking too long. The third theme related to the reason why women faked orgasm was because it was taking them too long to reach a real orgasm. They were worried that their partner would think something was wrong with them or they felt pressured by their partner continually asking, “Have you come yet?” Some examples of quotes from women describing how they felt they were taking too long included:

It’s more like I felt that I should reach an orgasm sooner, more than because he wasn’t going to give me one cause he’s doing the same things he usually does [when I have an orgasm] but sometimes it’s just not as fast. I feel like sometimes it takes me a lot longer to have an orgasm, but I don’t know why. I feel bad when it takes me a long time ‘cause then I think he feels bad too. (Delilah)

I feel like the guy is going to think that something is wrong with me if I don’t have an orgasm. I don’t want him to think that he just had sex with a weirdo after he finishes. He would ask me if I had orgasms, and I hated that. He did it all the time during sex, it made pleasure really difficult to focus on. (Bella)

He didn’t know what he was doing. The fourth theme related to reasons women gave for why they fake an orgasm was because the man they were with did not seem to know what he was doing in terms of stimulating her. Women described how they were sure they would not be reaching a real orgasm with him and therefore felt their only option was to fake an orgasm. Examples of this included:

I think with the other guys [one night stands] it was different. They didn’t really know what they were doing. If I was going to be in a long-term relationship, it would be different, but I had no confidence in the men that I slept with. I would always be like “Eh, they’re never going to get it,” so I would fake because I’m pretty sure that I just wouldn’t have ever have an orgasm in those encounters. (Emma)

I think that the majority of the time I fake because they are not doing anything and I’m not getting very stimulated throughout the whole thing, so I just don’t feel like it’s going to happen no matter how long it goes. No matter how long we like had sex, I wasn’t going to have an orgasm. (Lily)

It’s expected. The fifth theme related to reasons why women faked orgasms included an expectation and pressure to have an orgasm with intercourse. All of the college women said they believed it was what needed to happen in order for sexual intercourse to be complete. Not only should he orgasm, but she needs to orgasm as well. Women described this expectation and pressure in the following examples:

Guys just think we should have an orgasm, so that’s how it happens. I think there’s a huge amount of pressure on many women to have an orgasm, so they end up faking. I wouldn’t say anyone is happy to fake orgasms because normally the situation that you find yourself in is not ideal. (Abigail)

I feel like guys expect you to orgasm, but among girls I feel like it’s an understanding that having an orgasm during sex is not quite so common. I kind of think that’s where faking comes from because girls all understand how difficult it can be to have an orgasm, but guys just don’t seem to get it. I think that they kind of like originate the pressure, but sometimes I feel like I internalize that and then like I feel like I need to have an orgasm. (Emma).

Feelings about Faking: Hers, Her Partner(s), and Her Friends

The third research question asked, “How do college women feel about faking and what is their sense of how their partner(s) and friends feel about women faking orgasm?” The 12 college women were asked a series of questions to explore their feelings about faking, as well as the feelings of those around them. Four unique themes were identified from the in-depth interviews. Many women stated that both she and her friends feel that faking orgasm was typical or normal behavior. Most admitted feeling guilty, but recognized it was necessary, and they described how they never discussed faking with their sexual partner because they knew his response would not be good. These are described below and are listed in Table 2.

I feel it is pretty normal. The first theme related to the women’s personal feelings about having faked orgasm and centered on how normal this behavior was within a sexual encounter. Women seemed to shrug their shoulders and be unphased about having faked and orgasm. Many were okay with faking orgasm and saw it as a useful tool in many sexual encounters. Examples of women’s description of their feelings include:

I don’t think I’m doing anything bad. Just kind of like neutral. I think it’s pretty normal though, and I don’t think that many women feel bad about faking, especially because they are faking to not hurt their partner’s feelings. (Delilah)

It’s so normal, almost like, it shouldn’t be so normal, it shouldn’t be something that I have to do, I just don’t even think about it. I feel like it’s become like I’m used to it. It is what it is. No one really puts any thought into it. I don’t care that I have done it personally, but it’s probably something that I should change at some point. (Emma)

I feel guilty, but it was necessary. The second theme focused on how women feel about having faked orgasm and involved guilt. However, these women also qualified these guilty feelings by saying that faking was necessary in many situations they found themselves in order for the sexual encounter to be considered a success. Some quotes from college women about these feelings include:

I feel bad that I didn’t address it, I mean lying is really what I feel guilty about, but I didn’t want to make either of them feel bad. Going back, I would definitely talk more about it ‘cause that’s super important but going back I was just too shy at the time to discuss it. (Irene)

It is disappointing to fake cause I didn’t feel like my needs were important enough to tell the guy about. It is always hard for me to say, “That wasn’t good for me.” I am also okay that I didn’t make them feel bad, but also I was lying so then you kind of feel guilty anyway. (Faith)

My sexual partner would feel deceived. The third theme to emerge related to feelings about faking orgasm involved her thoughts on how her sexual partner would feel if he knew. All of the college women indicated they were not willing to tell their sexual partner they had faked because they were sure he would respond in a negative way. All of them were sure he would feel lied to or deceived. This reaction was confirmed by a few women who shared that they had later told a sexual partner about faking, and his response was to feel lied to and deceived. Sample responses included:

I haven’t ever talked about that with any of my sex partners. That would be so awkward and embarrassing for both of us because he would feel like he couldn’t get me off. (Bella)

I don’t want it to come out that I have faked in the past, so it’s not something that I ever bring up with my partner. I don’t want to hurt my partner’s feelings, so I would not want to talk about it. I don’t think that guys like to think that anyone has ever faked with them because that would hurt their confidence, so they probably don’t want to bring it up either. (Kaylee)

My girlfriends feel it is perfectly okay. The fourth theme related to feelings about faking orgasm centered on her friends’ reactions to finding out they fake. The overwhelming response from her girlfriends was that it is perfectly normal behavior and not surprising. Women talked only about their female friends’ response; none of the women discussed how male friends would feel about her faking orgasm with a partner. Some example of friends’ feelings about faking orgasm include:

I talked to my friends before I met you for this interview. I texted everyone. And one of my roommates responded back, “Have I faked an orgasm? Do you mean am I a woman?!” We all just laughed because it’s so regular. I feel like with my friends it’s the norm, and that’s what a lot of women unfortunately do. (Abigail)

So the only people I’ve ever talked about this with are my current roommates. We talked about it and all of us kind of joked like, “Oh girl, have I faked? Hell yes I faked, who hasn’t faked?” But it never went really beyond that. They didn’t have any specific reaction when I told them that I personally had faked. I think that most women just assume that all other women are faking, too. (Charlotte).

Sexual Scripts

An examination of the transcripts revealed three overarching sexual scripts these college women internalized to explain their faking orgasm behavior. These sexual scripts include: 1) media messages of what “normal sex” looks like, 2) her orgasm is a reflection of his competence, and 3) her orgasm signals that sex is complete. These sexual scripts are discussed below. Sample quotes are included to highlight these scripts.

Media messages of what “normal sex” looks like. The first sexual script centered on the media. Many of the women didn’t know exactly where their ideas, reasons for, or feelings about faking orgasm originated from, but they were able to cite the media as playing a very important role. Women discussed movies, television shows, and pornography. Women found these various types of media to be influential in their sex lives and to offer them an understanding of what “normal sex” should look like. Some sample quotes describing women’s experiences with media are below:

I think in my experience the way that men and women act has a lot to do with porn and when they watch it. Like the women through the whole entire thing are having orgasms, multiple orgasms. I think that porn plays such a big role in the expectations today. If you have a partner that looks at porn and always thinks it’s going to be the reality of sex, then that’s going to cause a lot of problems in the relationship. (Delilah)

I haven’t really thought about where I learned about faking orgasm before. I don’t want to say TV shows because that’s cliché, but you know how sometimes on TV you might see stuff like that, I don’t know, but I feel like the women on the TV fake it. It’s not like I’ve ever really given it thought, though I guess, I think I would say TV was a pretty big influence. (Grace)

I think that porn is a big reason why women fake, the way that sex is shown makes men think that sex in real life is going to be different than it actually is. Even in movies and stuff, sex is like this amazing thing that everyone wants to have, like in American Pie, but in real life it’s not ever really that interesting. I think sex can be really boring, especially if the guy has no idea what do to with your body. (Faith)

Her orgasm is a reflection of his competence. The second sexual script centered around her orgasm as a reflection of him. Nearly every single woman interviewed had faked orgasm because they understood that her ability to reach orgasm assured him he was competent. This sexual script suggests these women have learned that sex is about him; it’s about assuring him of his abilities, his sexual prowess, and his manhood. All women described the pressure from their partner to orgasm in order to assure him of his competency. Some examples of this can be found below:

With my boyfriend I would fake because I didn’t want to make him feel bad. And I think the other two guys, I mean, maybe it was just kind of, I guess the best reason would be like societal expectations. I don’t know, I always kind of feel like it’s something that should happen during sex. Even if they didn’t make me orgasm, they did make me feel good in at least some way, and faking was kind of my way of like showing even if they really didn’t make me orgasm, they were doing something good. (Grace)

Um, honestly I fake for him, so as not to hurt their ego because men are babies. They couldn’t handle rejection, so I would basically just fake with them because I didn’t want to hurt their feelings or anything. Men need to feel strong, needed, and important. So, I think that giving a girl an orgasm is very important to their self-esteem; if they are unable to give a girl that orgasm it hurts their self-esteem and makes them feel like they aren’t as sexually powerful as they should be. (Bella)

I think that men kind of originate the pressure to have an orgasm. Sometimes I feel like I internalize that and then like I feel I need to have one. Guys always want you to have an orgasm. I think it makes them feel better and more accomplished and then they get the added bonus that their partner is having a good time too. (Emma).

Her orgasm signals that sex is complete. The third sexual script is that women felt like sex was not complete until she experienced an orgasm. Not only did women feel this way, many also proclaimed that their male partners felt similarly. This pressure to perform in order to make the sex act complete was a recurring sentiment. Some sample quotes are below about women feeling as though their orgasm signals to the male partner that he can now orgasm and then sex is complete:

I wanted it to be over, so faking the orgasm was how I decided that I wanted to end the sexual encounter. I learned that if I wanted to end sex, then I could fake orgasm because that was a signal to him that he should be finishing up soon. (Charlotte)

At the time I felt like an orgasm was supposed to happen and then that was sort of like the mark of the end of the experience so I should basically have one to finish things up. (Irene).

Discussion

This study sought to add to our understanding of how and why women fake orgasm. There have been previous studies on women who fake orgasm, however, several are decades old and/or include a very broad age range of participants. For a behavior that appears to be extremely common in sexually active women, and one that has been increasing among college women over the past 30 years [8], there is a need for more research to understand this phenomenon. This study focused exclusively on a younger cohort of educated, college women who had faked orgasm. It involved in-depth interviews in order to understand the context, reasons, feelings, as well as to determine the sexual scripts that are influencing faking orgasm among today’s young women.

Comparison of Findings to Previous Research

Frequency and context of faking orgasm. In terms of the frequency and context of faking orgasm, there was a great variation in how often these college women reported they had faked orgasms. Some women reported that they faked every single time they had sex, while others had only faked a few times in their life. Other researchers also found that there was a wide range in frequency of faking among their study participants. For example, Bryan’s (2001) [15] found that 20% of women faked orgasm during 20% of their encounters, Fahs (2014) [11] found that 45% of women report that they faked orgasm “regularly” during their sexual encounters, and Ellison (2000) [10] found that while most had faked less than 50 times; 10% of the women claimed to have faked orgasm between “150 and 10,000 times or wrote in another answer such as ‘countless’ or ‘a bazillion’” [10].

Most of the college women in this study discussed pretending to orgasm by using their bodies and lying if their partner asked them about having an orgasm. Many women described “putting on a show” for their male partner so that they would not suspect anything. This was a finding described in other studies of women faking orgasm (e.g., Bryan, 2001) [15]. In addition, they all discussed how faking orgasm typically occurred with sexual intercourse. Few women identified faking during oral sex. Earlier studies (e.g., Darling & Davidson, 1986; Wiederman, 1997) [9,14] also specifically explored women who faked orgasm and found this to be typical.

Two findings related to frequency and context were unique to this research. First, the college women in this study reported that they got better at faking over time. Many noted that the longer they faked, the more convincing they felt they became. And related to this, once they experienced a real orgasm, they felt their ability to convincingly fake future orgasms improved considerably. The other unique finding that has not been discussed in previous research related to the context was that these college women described faking orgasm at the beginning of a relationship. All of the women faked when they were “less close” with their partners and had “less emotional attachment.” For some women (those who had only one-night stands), this pattern continued, but for those who had long-term boyfriends, they stopped faking as the relationship progressed.

Reasons for faking orgasm. In terms of the reasons why women fake, almost every woman interviewed agreed that they faked orgasm in order to save their partner’s feelings. The women were more concerned with their partner’s ego than with their own pleasure during the sexual experience. This finding was similar amongst nearly all of the previous research on reasons why women fake [7, 10, 13, 15, 16, 19, 20, 21]. For many women, it was reported that it was easier to fake and “protect his feelings” than to tell him the truth and deal with an uncomfortable conversation. Other reasons women cited for faking were similar to previous studies noted earlier including being uncomfortable, worrying she was taking too long, and feeling the pressure to orgasm.

Unique findings in this study related to reasons women fake orgasm included college women talking about experiencing physical pain, lack of concern for maintaining the relationship, and because he did not know what he was doing. In this study, many of the women cited feeling physically uncomfortable (not just emotionally uncomfortable) and cited this as a reason for faking orgasm in order to have the sexual encounter stop. Few of the previous study [15] discussed that women fake orgasm because of physical discomfort.

In addition, many of the previous studies found that a common reason why women fake was to keep the relationship intact [10, 12, 17, 20]. In the current study, not one of the college women talked about this as a reason for faking an orgasm, which raises questions about today’s college women and their focus on long-term commitments.

Another unique theme noted by college women in this study to explain their reason for faking was because their partner did not know what he was doing. The women who discussed this felt as though they would never experience an orgasm with their partner and therefore faked orgasm. These women were faking orgasm in order to end the encounter, and this is where there are similarities to the other studies [10,11,13,15,18]. These previous studies found that women would fake orgasm to end the sexual encounter, however it was for a variety of other reasons such as being tired, bored, or uncomfortable, not that the male partner didn’t know what he was doing. This reason for faking could be explained in large part because of the young age of the female respondents, who in turn may have young, sexually inexperienced partners who do not know what is involved in pleasing their partner.

Feelings about faking. Several studies have investigated how women feel about faking orgasm and women have suggested their male partner would be hurt to know [10, 11, 18], but asking about how their friends feel about their faking behavior has not been a focus of past studies. In this study, the college women were divided in their own feelings about having faked orgasm. On the one hand, many of the women felt as though their behavior was normal and nothing they were concerned about; on the other hand, many women felt guilty about faking orgasm, but they also justified their actions as necessary. These women knew that faking may not have been the right thing to do, but they could justify why it was done. In addition, most of the college women mentioned that they didn’t tell their partner about faking orgasm because they did not want him to have a negative response.

A unique finding in the study was the reaction of her friends. The college women interviewed for this study all described their female friends as having a supportive view of faking orgasm. Many of their college girlfriends shared that they do the same thing, and that they also believe faking orgasm is a very normal behavior.

Sexual scripts. The finding that many college women discussed the role of the media in defining what “normal sex” is and what female orgasm is supposed to look like was unique. Few of the previous studies [11] examined women’s sexual scripts or the variety of societal pressures that are placed on women. All the women in this study described the important role of the media in shaping their understanding of sex and orgasm. The college women participating in this study mentioned either learning about orgasms or faking orgasms through some sort of media such as pornography, movies, or television. These women mentioned having a fantasy version or unrealistic idea of what “normal sex” is “supposed” to look like due to the media influence in their younger years.

The second script that arose from the interviews with college women is the notion that her orgasm serves as a reflection of her partner. Previous studies have also described this need for women to assure her partner of his competency. [26, 27] discuss how it becomes obligatory for women to have an orgasm. A large goal of her orgasm is not about her pleasure; rather it is about making sure her male partner is happy and confident about his own sexual abilities. This belief system further ingrains the sexual double standard that the male orgasm is for pleasure while the female orgasm is for performance. Other studies [11, 15] found that many women faked because they felt that they needed to show their partner that he is doing a good job and give him the impression that they are enjoying sex as much as he is.

Finally, the third script to emerge from the interviews was the notion that female orgasm signals that sex is complete. Many of the women discussed faking orgasm in order to end the sexual experience. This is similar to other studies such as [26] in which they noted that the woman’s orgasm is the “goal of sex.” Similar to the current study, the female orgasm is seen as what needs to happen for the sex to be complete.

Limitations

As with all qualitative studies, the findings of this study cannot be generalized. There were several limitations to this study beginning with a small sample size and being limited by participants who were willing to be interviewed for the study. In-depth interviews were completed with college women who volunteered and therefore may be more open and comfortable with talking about faking orgasm. The interviews relied on the participants’ ability to recall as well as their perceptions of events, which could not be verified. This study was also limited to students who attended one public university in the northeastern U.S. and all shared the same race. A more diverse group of participants may impact the findings. Factors such as women’s race, socioeconomic status, culture, or religious orientation could all play into women’s propensity towards faking orgasm, along with their willingness to discuss such matters.

Future Research

The prevalence of faking orgasm and the reasons why women fake have been the focus of many studies, however, as noted earlier, it is one of the “least well-studied human sexual behaviors” [7]. In relation to the current study, it would be beneficial for future studies to conduct more in-depth investigations focusing on specific aspects of the current study. Future studies could focus solely on the context of faking, reasons for faking, or feelings a woman or her friends might have about faking orgasm. It would also be beneficial to look at the deeper meaning of faking orgasm in terms of the sexual scripts.

It is important to note that all of the participants in this study ranged in age from 19 to 22 years old. This sample was chosen because of the interest in examining young women’s experiences. Recognizing that faking orgasm is increasing among this age group, it is important to understand why. It is useful to examine this age group to understand how society, media, self-image, and male partners all affect their faking behavior. Other studies could compare how younger women differ from older women in their attitudes, beliefs, and influences. Future research could also compare women based on other factors such as ethnicity, race, religion, socioeconomic factors, and sexual orientation. The participants for this study were all women who shared their experiences of faking orgasm with male partners. Further research could examine how women in same sex relationships experience faking orgasm.

Suggestions and Recommendations

The frequency with which women fake orgasm, and the pressure women feel from their partners to reach orgasm provides several suggestions and recommendations for sexuality education. First and foremost, there is greater need for education of basic anatomy, as well as opportunities for discussion of what students see as the purpose of sex. It appears that for many of the women interviewed in this study, sex involves feelings of pressure, and involves an act or performance in order to prove (mostly to the male partner) that the sexual encounter was a success and that she is normal and okay. Better sexuality education is needed for both men and women in order to have a more realistic understanding of sexual function and the role of pleasure.

Second, there is a need for media literacy and a discussion of the role of pornography specifically in shaping ideas about sex. Many of the women felt their faking behavior was influenced by what has been portrayed in pornography, as well as in television shows and in movies. An opportunity to discuss the reality of sexual encounters versus the image portrayed in the media would be useful.

Third, there is a need for greater permission for couples to talk about their sexual response, their desires, motives, what feels good, and what they need. The college women interviewed for this study made it clear that sex is not something you can or should talk about with even your most intimate partner. Better role models for communicating and greater emphasis in sexuality education classes on how to approach topics of sexual pleasure are needed to give both women and men permission to talk with their partner.

Conclusion

This research looked at an understudied aspect of women’s sexuality – faking orgasm – from a qualitative perspective. It focused on young women who fake in order to understand the reasons, context, and influences for such behavior. Among these women’s circles of friends, faking orgasm is considered commonplace and normal. Faking orgasm is not new; however, it appears that more women are faking than in prior generations. For college women, faking behavior has increased dramatically over the past 30 years – from less than half to over three-quarters of college women today saying they have faked an orgasm [8]. This increase in faking seems counter-intuitive as the emergence of the third wave of feminism has worked to empower women, and women are becoming much more aware of their own pleasure and their own bodily autonomy. There are a wide variety of reasons women give for faking, and by delving into these reasons, the sexual scripts that women are raised to believe can be better understood. By deciphering these scripts that lead to faking orgasm, researchers can further understand societal pressure, self-pressure, and partner pressure, and work to change that.

This study was unique in that it used a qualitative methodology to investigate the experiences of college women who had faked orgasm in the context of heterosexual relationships. Women faking orgasm appears to be normalized amongst their peer group. Interviews with these 12 college women revealed many themes and suggested that women have similar experiences when it comes to faking orgasm and their sexual scripts. By understanding the scripts that young women who fake orgasm hold onto, along with the context and reasons for faking orgasm, we can further deconstruct the societal implications tied to what “normal sex” is “supposed” to look like and help them have a more honest and pleasurable sexual experience.

References

  1. Reiner R (1989) When Harry met Sally. MGM Home Entertainment.
  2. David L, Seinfeld J (1993) Seinfeld/The Mango [Episode 65]. In Seinfeld. NBC.
  3. Daniels G, Poehler A, Schur M, Wittles H (2011) Parks and Recreation/Ron and Tammy: Part II [Episode 34]. In Parks and Recreation. NBC.
  4. TMZ (2016) Caitlyn Jenner: Watch me fake an orgasm. I am Cait. TMZ: Los Angeles, California.
  5. Breslaw A (2017, October 11). 7 Sad but true reasons women fake orgasms. Cosmopolitan.
  6. Azodi M (2010, October 26). The crazy way I faked it. Cosmopolitan.
  7. Cooper EB, Fenigstein A, Fauber RL (2014) The Faking Orgasm Scale for Women: Psychometric properties. Archives of Sexual Behavior 43: 423-435. [crossref]
  8. Caron SL (2021) The sex lives of college students: Three decades of attitudes and behaviors. Orono, ME: Maine College Press.
  9. Darling CA, Davidson JK (1986) Enhancing relationships: Understanding the feminine mystique of pretending orgasm. Journal of Sex & Marital Therapy 12: 182-196. [crossref]
  10. Ellison C (2000) Women’s sexualities: Generations of women share intimate secrets of sexual self-acceptance. Publisher’s Group West.
  11. Fahs B (2014) Coming to power: Women’s fake orgasms and best orgasm experiences illuminate the failures of (hetero) sex and the pleasures of connection. Culture, Health & Sexuality 16: 974-988. [crossref]
  12. Hite S (1976) The Hite report: A nationwide study of female sexuality. New York: Seven Stories.
  13. Muehlenhard C, Shippee S (2010) Men’s and women’s reports of pretending orgasm. The Journal of Sex Research 47: 552-567. [crossref]
  14. Wiederman MW (1997) Pretending orgasm during sexual intercourse: Correlates in a sample of young adult women. Journal of Sex & Marital Therapy 23: 131-139. [crossref]
  15. Bryan TS (2001) Pretending to experience orgasm as a communicative act: How, when, and why some sexually experienced college women pretend to experience orgasm during various sexual behaviors (Unpublished doctoral thesis). University of Kansas.
  16. Braun V, Clarke VE, Rogers C (2014) “It feels so good it almost hurts”: Young adults’ experiences of orgasm and sexual pleasure. Journal of Sex Research 51: 503-515. [crossref]
  17. Kaighobadi F, Shackelford TK, Weekes-Shackelford VA (2011) Do women pretend orgasm to retain a mate? Archives of Sexual Behavior 41: 1121-1125. [crossref]
  18. Lafrance MN, Stelzl M, Thomas EJ (2016) Faking to finish: Women’s accounts of feigning sexual pleasure to end unwanted sex. Sexualities 20: 281-301.
  19. Mialon HM (2012) The economics of faking ecstasy. SSRN Electronic Journal 50: 277-285. [crossref]
  20. Roberts C, Kippax S, Waldby C, Crawford J (1995) Faking it: The story of “ohh!” Women’s Studies International Forum 18: 523-532.
  21. Fahs B (2011) Performing sex: The making and unmaking of women’s erotic lives. Albany, NY: SUNY Press.
  22. Harris EA, Hornsey MJ, Larsen HF, Barlow FK (2019) Beliefs about gender predict faking orgasm in heterosexual women. Archives of Sexual Behavior 48: 2419-2433. [crossref]
  23. Potts A (2000) Coming, coming, gone: A feminist deconstruction of heterosexual orgasm. Sexualities 3: 55-76.
  24. Future of Sex Education Initiative (2012) National sexuality education standards: Core content and skills K-12. Special issue of the Journal of School Health 1-42.
  25. Casey E, Masters NT, Morrison DM, Wells EA (2013) Sexual scripts among young heterosexually active men and women: Continuity and change. Journal of Sex Research 50: 409-420. [crossref]
  26. Lavie-Ajayi M, Joffe H (2009) Social representations of female orgasm. Journal of Health Psychology 14: 98-107. [crossref]
  27. Weiss S (2017) 4 super real reasons women fake orgasms.
  28. Maxwell JA (2005) Qualitative research design: An interactive approach (2nd Edition). Thousand Oaks, CA: Sage Publications.

Reflections on Women’s Medical Autonomy in the 21st Century

DOI: 10.31038/AWHC.2023622

Editorial

One of the fiercest and most pragmatic of women’s battle was their right of medical autonomy vis-a- vis full information and right of acceptance/ refusal of any treatment offered. One of the most celebrated of these medico-legal battles and one which proved a page-turner was the case of Montgomery (Appellant) v Lanarkshire Health Board (Respondent) [2015] UKSC 11. It is time to take a reality check on the situation arising in this case, for it is becoming amply clear that in daily obstetric practice, women’s rights are far from being respected for one reason or another.

Montgomery (Appellant) v Lanarkshire Health Board (Respondent) [2015] UKSC 11 revolved about a pregnant mother’s request for an elective caesarean section was completely rejected by the obstetrician. The mother, a PhD graduate, repeatedly and persistently informed her obstetrician about her fears of a vaginal delivery. Her request for a CS was not unreasonable, for she, a primigravida, would be delivering a baby presenting by the breech. In addition,

(i)She was of a short stature.
(ii)She was a diabetic.
(ii)She was carrying a clinically large baby (4.25kg).

Subsequently, at birth the baby experienced severe shoulder dystocia and developed cerebral palsy of the spastic quadriplegic dyskinetic form consistent with underlying Hypoxic-Ischaemic Encephalopathy. At the Appeals Court, the defendant obstetrician was deemed negligent and the Appellant was awarded the sum of £5.25 million in damages. The ruling is widely held to have displaced the previous “Bolam test” in matters of consent.

There is no doubt that the hefty sum awarded in Montgomery was impressive as was indeed the now doubly underlined warning that that the doctor does not know all and the patient must be informed of all that has to happen to him/her and has the right to accept or reject any medical treatment proposed. This principle of disclosure in itself had been long a-brewing, but Montgomery pushed it to the fore not only in the Anglo-Saxon world but also across the ocean. Not that, anyone needed to be advised to explain managements to patients and listen to their side of the coin.

There was and is a danger with Montgomery that the medical practitioner just listens to what the patient wants and simply concedes. This is not a rare occurrence especially with caesarean sections in private practice. That is not the spirit of the Montgomery judgement which, above all, requires that the physician explains the suggested treatment and any alternatives to it, the pros and the cons, what potential complications may be entailed etc. This is Montgomery and it seeks to elicit the charity, the compassion and the ‘loving father image’ rather than the now rejected paternal aspect of “I know best”. A loving father explains to a child and helps in the right choice. In the great majority of cases, medical practitioners across all disciplines are cut of the “loving father” cloth, but, by no means, does this apply universally so.

I have recently been greatly disturbed by a patient of mine, whose daughter in Germany had had a breech presentation and was repeatedly and forcefully refused a caesarean section. The obstetrician could not be a person well versed with recent obstetric developments nor with modern medical ethics in general. Having confirmed all the details, I was both much amazed and greatly pained at such dictatorial remnant behaviour in 2023. Perhaps this is even commoner than I thought. And in this case, we speak of a woman who was tertiary educated – a PhD, in fact. Imagine what an uneducated woman, a non-English speaker or a refugee goes would to through, at this obstetrician’s hands and his ilk.

The battle for woman’s rights is by no means over. I firmly believe that before such rights can be fought for, their existence must surface, be noted and registered. Taking obstetric care as one example, the mode of delivery, especially and particularly in complicated cases, must be actively discussed with the parents, with an end scope of truly answering their questions. If the mother’s choice is for a particular mode of delivery, the only solid element to bring out and defend against would be maternal and child safety. In a breech delivery, for example, the greatest potential danger in a vaginal delivery, would be fetal intra-partum obstruction with subsequent intra-partum hypoxia and its resultant damage such as cerebral palsy. The inherent surgical and anaesthetic risks in a modern-day caesarean section especially with regional anaesthesia would be far less than the previous situation as discussed.

In Medicine, where one must practice secundum artis, depending on challenge and circumstance, no one can write a book of magical solutions. However, as modern medicine broadens its scientific horizons and new legal and ethical reins regulate behaviour, it is now clear that just as important as the tenets of science that we must be informed about, are the obligations of behaviour, imposed by the continuously evolving principles of law and medical ethics.

Repeated Traumatic Brain Injury is Associated with Neurotoxic Plasma Autoantibodies Directed against the Serotonin 2A and Alpha 1 Adrenergic Receptors

DOI: 10.31038/EDMJ.2023722

Abstract

Objectives: Traumatic brain injury (TBI) was associated with increased plasma agonist autoantibodies targeting the serotonin 2A receptor. Repeated TBI exposure is associated with high risk for neurodegenerative and neuropsychiatric complications. Here we tested a hypothesis that repeated TBI is associated with plasma agonist autoantibodies targeting more than one kind of catecholamine G-protein coupled receptor.

Methods: Protein-A affinity chromatography was used to isolate the IgG fraction of plasma in forty-two middle-aged and older adults who had experienced one or more TBI exposures. The Ig (1/40th dilution=7.5 ug/mL) were tested for neurotoxicity in mouse neuroblastoma cells using an acute neurite retraction assay indicative of Gq11/IP3/Ca2+ and RhoA/Rho kinase signaling pathways’ activation. Three different linear synthetic peptides corresponding to the second extracellular loop of the alpha 1A, alpha 2A or serotonin 2A receptors were used as target antigen in different enzyme-linked immunoassays. The second extracellular loop receptor peptides themselves (alpha 1A, alpha 2A) or a fragment (serotonin 2A) were tested for ability to prevent Ig-induced neurite retraction.

Results: Patients who had experienced either repeated TBI (N=10) or a single TBI with a co-morbid autoimmune disease (N=5) were significantly more likely to harbor neurotoxic plasma autoantibodies targeting both alpha 1 adrenergic and serotonin 2A receptors vs. patients having only a single TBI. Ig-induced neurotoxicity was significantly prevented by co-incubation with either 850 nM prazosin (alpha 1 adrenergic receptor) and/or 500 nM M100907 (serotonin 2A receptor) antagonists. Alpha 1 adrenergic receptor and serotonin 2A receptor Ig immunoreactive level and titer were significantly increased in repeated TBI and single TBI/autoimmune patients (N=7-8) compared to age-matched TBI patients without neurotoxic plasma Ig (N=4). SN.8, a linear synthetic peptide corresponding to a conserved region of the second extracellular loop (ECL) of the serotonin 2A receptor completely prevented neurite retraction induced by repeated TBI plasma Ig. A repeated TBI patient harboring alpha adrenergic receptor AAB alone experienced prospective steep decline in cognitive function over two years.

Conclusions: Repeated TBI and TBI with associated autoimmunity harbored more than one kind of neurotoxic catecholaminergic agonist GPCR autoantibody each associated with high risk for steep rate of cognitive decline. Specific immunoassays using the second extracellular receptor loop as target antigen are needed to detect each specific different GPCR autoantibody. A fragment of the second ECL of the serotonin 2A receptor (SN.8) neutralized Ig-induced neurotoxicity in repeated TBI or TBI with associated systemic autoimmunity.

Introduction

Traumatic brain injury (TBI) contributes to substantially increased global disability including the increased risk for later occurrence of major depressive disorder, Parkinson’s disease or dementia [1]. United States military veterans of the conflicts in Afghanistan and Iraq who sustained repeated TBI exposures are also likely to suffer with chronic post- traumatic stress disorder (PTSD). Certain symptoms of chronic PTSD overlap significantly with those of TBI including mood changes (anxiety, depression, suicidal ideation), and altered cognition (attention deficits, memory impairment). A distinct subset of chronic PTSD symptoms (nightmares, panic attacks) involving hyperarousal suggest dysregulated norepineprhine and epinephrine signaling on alpha 1-adrenergic receptors expressed in the brain and sympathetic nervous systems [2].

G-protein coupled receptors are highly druggable treatment targets, and prazosin, a selective alpha1 adrenergic receptor antagonist is effective for the treatment of nightmare disorder in chronic PTSD [3]. Traumatic brain injury and its later neurodegenerative sequelae (major depression, Parkinson’s disease, dementia) was previously associated with increased plasma agonist autoantibodies targeting the serotonin 2A receptor [4,5]. Here we tested a hypothesis that repeated exposures to traumatic brain injury promotes persistent neuroinflammation leading to the development of humoral autoantibodies targeting two, related catecholaminergic GPCRs, i.e. the serotonin 2A and the alpha 1A adrenergic receptors. We used immunoassays specific for the second extracellular loop regions of the alpha1 A receptor and serotonin 2A receptors to test for co-occurrence of these catecholaminergic receptor- targeting autoantibodies (AAB) and its association with clinical outcomes in 42 middle-age and older adult TBI-sufferers.

Methods

TBI Patients

Informed consent for the local Institutional Review Board-approved studies was obtained from all participants prior to blood drawing or cognitive testing. The participants belonged to two different cohorts, A and B. Cohort A included patients age 50 years or older (N=35) among whom approximately two-thirds of patients were previously reported to harbor plasma autoantibodies targeting a second extracellular loop region of the serotonin 2A receptor [4]. Here we report additional bioassay results in thirty of thirty-five consecutively-enrolled patients from cohort A, i.e. neurite retraction in mouse neuroblastoma cells induced by plasma IgG fraction; and whether plasma IgG-induced neurite retraction was significantly reduced (70% or greater) by the presence of either a highly selective serotonin 2A receptor antagonist (M100907) or a selective alpha 1 adrenergic receptor (prazosin) antagonist. Cohort B is comprised of younger patients age 40-60 years who suffered either a single or repeated TBI exposures. In Cohort B we not only tested Ig for bio-assayable 5-HT2AR-like and alpha 1AR-like neurite retraction, but also for immunoreactivity targeting the second extracellular loop of the alpha 1AR (and/or 5HT2AR).

TBI Exposure

All participants in Cohort A experienced direct force traumatic brain injury. Cohort B participants included patients who had experienced either direct force (n=6), blast TBI (n=5) or both types of TBI (n=1). In nearly all cases, the TBI exposure was judged to have been mild.

Peptides

QN.18, PP.15 (PAPEDETICQINEEP), and RT.18 (RQPDAGAAYPQCGLNDET) are linear synthetic peptides corresponding to the second extracellular loop of the human 5HT2AR, alpha1a AR or alpha 2a AR, respectively. SN.8 (SCLLADDN) is comprised of a subregion of the human 5HT2AR involved in mediating receptor activation All peptides were synthesized at Lifetein, Inc (Hillsborough, NJ). The peptides had purity ≥ 95% and were stored under desiccated conditions at -40 degrees C prior to use. On the day of an experiment an aliquot of lyophilized peptide was dissolved in sterile, phosphate buffered saline or deionized water prior to immunoassay or bioassay.

Protein-A Affinity Chromatography

Protein-A Affinity Chromatography was carried out as previously reported [4]. The resulting IgG fraction obtained from TBI patient plasma was stored at 0-4 degrees C prior to bioassay and immunoassay.

Mouse N2A Neuroblastoma Cells

Mouse N2A Neuroblastoma Cells were obtained from the American Type Culture Collection (Rockville, MD).

N2A Neurite Retraction Bioassay for 5HT2AR-like and Alpha 1AR-like Neurotoxicity

We used a previously reported acute neurite retraction assay in cultured mouse N2A neuroblastoma cells [6] to test the IgG fraction of plasma from adult TBI patients for autoantibodies (AAB) causing significant acute neurite retraction. A selective serotonin 2AR (M100907) and alpha 1AR (prazosin) antagonists was each incubated (separately) with TBI patient IgG to test for significant inhibition (>/=70%) of the AAB-induced N2A neurite retraction.

Enzyme Linked Immunoassay

Enzyme Linked Immunoassay were performed as previously described using the QN..18 second extracellular loop of the 5HT2AR [5] as the target peptide antigen. For the alpha 1AR and alpha 2AR immunoassays, the respective specific second extracellular loop peptide for each receptor PP.15 (alpha 1AR) or RT.18 (alpha 2AR) was substituted in place of QN 18.

Rat Plasma, Protein-G Affinity Chromatography, Enzyme Linked Immunoassays

Rat Plasma, Protein-G Affinity Chromatography, Enzyme Linked Immunoassays plasma was obtained from 25-week-old male spontaneously hypertensive rats (SHR) as previously reported [7]. The IgG fraction was isolated using protein-G affinity chromatography as previously reported [8]. Enzyme linked immunoassay was performed as previously described [7] using either the 5HT2AR second extracellular loop peptide QN.18 or the alpha 1A adrenergic receptor second extracellular loop peptide PP.15 as target antigen.

Statistics

Statistical analysis was performed using unpaired Students’ t-test and Fischer’s exact test.

Results

Co-occurrence of Plasma Alpha 1 Adrenergic- and Serotonin 2A-receptor Autoantibodies in TBI

Table 1 shows the clinical characteristics in a subset of 7/30 TBI patients (from Cohort A) whose plasma IgG had the properties of both alpha1AR targeting and serotonin 2AR target AAB. All seven patients had experienced either recurrent TBI or a single TBI exposure in the setting of having an underlying autoimmune disorder. The dose-response curves of IgG-induced N2A neurite retraction in these seven patients was compared to that of twenty additional patients who had experienced a single TBI and harbored either serotonin 2AR-targeting AAB alone or a lower level of uncharacterized AAB activity (Figure 1). Mean potency (neurotoxicity) in a 1/80th dilution from repeated TBI or autoimmune + single TBI plasma IgG significantly exceeded (P < 0.05) potency in an identical concentration of IgG from single uncomplicated TBI (Figure 1).

Table 1: Clinical characteristics in a subset of older adult TBI patients (Cohort A) who harbored both 5HT2AR-like and Alpha 1 AR- like bioactive Ig or Alpha 1 AR-like Ig alone.

tab 1

rTBI: Repeated Traumatic Brain Injury; Sys: Systemic; PTSD: Post Traumatic Stress Disorder; cognitive dysfunction.

fig 1

Figure 1: The indicated dilution of protein-A eluate fraction of plasma was incubated with mouse neuroblastoma cells and acute neurite retraction was determined as previously reported [ ]. Results are the mean ± SE of two or more measurements. *P<0.05: mean N2A neurite retraction in Ig from seven repeated or autoimmune TBI plasma significantly exceeded level in Ig from twenty patients who suffered a single TBI, not complicated by an autoimmune disorder.

Younger TBI patients many of whom are US military combat veterans having served in Afghanistan or Iraq they had a high rate of repeated TBI exposure and also suffer from post-traumatic stress disorder, whose symptoms overlap with those of TBI (e.g. anxiety and depression). The clinical characteristics in a subset of younger TBI patients (Cohort B) is shown in Table 2 with comparison to Cohort A. Cohort B patients had significantly lower mean age compared to Cohort B patients (Table 2). Six of twelve Cohort B patients tested had (N2A neurite retraction bioassay) evidence for both plasma 5HT2AR and alpha 1AR-like AAB (Table 2) including four patients who experienced repeat mild TBI and two patients who had a single TBI and a coexisting autoimmune disorder (not show in Table 2). The prevalence of chronic PTSD was quite high in Cohort B (75%) but it did not differ significantly from the PTSD prevalence in Cohort A (42%).

Table 2: Clinical characteristics in nested cohort of twelve younger TBI patients (Cohort B): comparison to subset of Cohort A patients harboring 5HT2A and/or alpha 1A receptor AAB.

tab 2

*Four patients had repeated TBI exposures, two patients had single TBI and an autoimmune disorder; ^ six patients harboring both 5HT2A and alpha1- receptor autoantibodies, one patient with only alpha1- receptor autoantibodies. AAB- autoantibodies, PTSD- post traumatic stress disorder. Alpha 1AR- alpha 1 adrenergic receptor.

When the results from Cohort A (N=30) and B (N=12) were combined, a striking association was evident between the presence of 5HT2AR and/or alpha 1AR-targeting bioactive Ig (vs. 5HT2AR Ig alone vs. neither AAB) and either recurrent TBI (7/12 vs 0/15 vs. 0/15; P=0.0016) or single TBI patients having a comorbid systemic autoimmune disorder (P=0.003) (Table 3). These data suggest that repeated TBI exposure carries an equivalent high risk as systemic autoimmunity for development of agonist autoantibodies targeting both 5HT2A and alpha 1A receptors (Table 4).

Table 3: Risk factors associated with combined presence of 5HT2AR and/or Alpha 1AR agonist Ig neurotoxic bioactivity in mouse N2A neuroblastoma cells.

tab 3

^ N=1 patient with recurrent TBI had alpha 1 AR AAB alone

Table 4: PTSD symptoms in Cohort B TBI patients: relation to AAB status

tab 4

^excluding obstructive sleep apnea

Agonist autoantibodies targeting each kind of receptor have been previously reported to be associated with an increased risk for dementia [9,10]. Neurodegenerative disease (dementia, cognitive dysfunction and/or Parkinson’s disease) was highly prevalent (5/7) among older Cohort A patients harboring both 5-HT2AR and/or alpha 1AR AAB (Table 1). In two such patients with recurrent TBI and chronic PTSD (Pt 4 and 6; Table 1) a 7.5 mg/mL (50 nanomolar) concentration of the plasma Ig caused potent 70-75% neurite retraction after 5 mins exposure in N2A cells (Figure 2A). Neurite retraction was substantially prevented by co-incubation with an 850 nanomolar concentration of prazosin (Figure 2A). Plasma Ig in two patients having Parkinsons’ disease (PD) and/or dementia (Pt 1 and 4; Table 1) mediated dose-dependent potent N2A neurotoxicity (i.e. neurite retraction) (Figure 2B). One of the patients (Patient 4, Table 1) who was observed prospectively (over two years) progressed from near normal baseline cognition to clinical dementia, in serial St. Louis University Mental Status testing (Figure 2c). The Patient 4 plasma alpha 1AR-targeting AAB was not detected in an enzyme-linked immunoassay specific for the second extracellular loop of the 5HT2AR (data not shown in Figure 2).

fig 2

Figure 2: A) Protein A- eluate (7.5 mg/mL) from two patients suffering with recurrent TBI, PTSD (Pts 4&6, Table 1) induced acute neurite retraction in N2A mouse neuroblastoma cells. The Ig- induced neurotoxicity (neurite retraction) was substantially prevented by co-incubating cells together with 850 nM concentration of the alpha 1 AR antagonist prazosin. B) Dose-response curves of neurotoxicity in the protein-A eluate fraction of plasma from TBI/PTSD (Pt 4) or TBI/Parkinson disease (Pt 1, Table 1). C) Pt 4 (TBI/chronic PTSD) harboring alpha1AR agonist Ig tested negative in 5HT2AR enzyme linked immunoassay, but experienced substantial 2-year decline in St. Louis University Mental Status (SLUMS) examination score. Dashed line indicates the cutoff score below which the score is indicative of dementia.

Taken together, these data suggested that younger recurrent TBI patients (including those in Cohort B) may be at substantially increased risk for the future development of clinically significant decline in cognitive function. An immunoassay specific for 5HT2AR Ig alone may not suffice to identify all such high- risk TBI patients.

The bioassay (N2A neurite retraction) data in all 42 Cohort A and B patients is summarized here. Twenty-one of 42 patients (50%) had IgG neurotoxicity which was significantly prevented by co-incubation of N2A cells with 200-500 nanomolar concentration of the highly selective 5HT2AR antagonist M100907 (Figure 3A). Fourteen of 42 patients (33%) had IgG neurotoxicity which was incompletely neutralized by M100907 and was not further characterized (Figure 3B). Eleven of 42 patients (26%) had IgG which was (blocked by M100907) and by co-incubation with 850 nanomolar concentration of the selective alpha 1 adrenergic receptor antagonist prazosin. Six patients (14%) demonstrated a low level of neurotoxicity in plasma which was not further characterized. One of 42 patients (2%) tested had an IgG which was solely inhibited by prazosin (alpha 1AR-like), but not by M100907 (5-HT2AR-like). The total exceeds 100% because patients (26%) harboring more than one Ig were counted twice. In an immunoassay specific for the second extracellular loop of the 5HT2AR, patients having 5HT2AR and/or alpha 1AR agonist-like neurotoxic bioactivity (N=12) had significantly higher level of 5HT2AR immunoreactivity compared to TBI patients (N=14) in whom 5HT2AR- or alpha 1 AR-like neurotoxicity could not be confirmed in bioassays (P=0.024) (Figure 3B). These data are consistent with a prior report that human pathologies’ 5-HT2AR Ig-induced neurotoxicity was highly correlated with plasma 5HT2AR immunoreactivity [5] using an enzyme linked immunoassay having QN..18, the entire second (5HT2AR) extracellular loop peptide, as target antigen.

fig 3

Figure 3: A) Neutralization of neurotoxicity in 21 of 42 TBI protein-A eluates by 200-500 nM concentration of the highly selective 5HT2AR antagonist M10097. B) Incomplete neutralization of neurotoxicity in 14 of 42 TBI protein A eluates by 200-500 nM concentration of M100907; C) binding to 5-HT2AR second extracellular loop peptide in protein-A eluates from TBI patients displaying (N=12) or not displaying (N=12) M100907-inhibitable neurotoxicity.

Sleep disorders including trauma nightmares are common in recurrent TBI patients suffering with chronic PTSD. Neutralization of combined (5HT2AR and alpha 1AR) plasma Ig-induced neurotoxicity in a representative patient with recurrent TBI and nightmare disorder is shown in Figure 4. Prazosin, an alpha 1 adrenergic receptor antagonist, is FDA approved to treat hypertension, and was effective in lessening the symptoms in traumatic nightmare disorder in combat veterans [3]. Here we conducted an exploratory analysis of whether alpha 1 AR agonist AAB may be associated with traumatic nightmare disorder in combat veterans suffering recurrent TBI.

fig 4

Figure 4: Ninety-three percent neutralization of neurotoxicity in the protein A eluate of plasma from recurrent TBI (Pt 5, Table 1) having traumatic nightmare disorder by 200 nanomolar concentration of the highly selective 5HT2AR antagonist M100907; eight-two percent neutralization by 850 nanomolar concentration of the specific Alpha 1AR antagonist prazosin. Results are mean ± SEM.

The prevalence of sleep-disorder (excluding obstructive sleep apnea) in Cohort B patients harboring alpha 1AR AAB vs Cohort B patients not harboring plasma alpha 1 AR AAB was increased (5/8 vs 1/4), but the sample size was too small and lacked sufficient power to detect a statistically significant difference. Of interest, Patient 4 harboring plasma alpha 1AR AAB was prescribed prazosin which provided symptomatic relief from traumatic nightmares. Yet despite regularly filling the prazosin prescription, he still experienced a substantial prospective decline in cognitive function over a two-year period. This observation may be consistent (in part) with the relatively short half-life of prazosin’s action and reports that alpha 1AR Ig mediates long-lasting receptor activation.

Mouse neuroblastoma N2A neurite retraction induced by the protein A eluate from a recurrent TBI patient harboring both alpha 1AR and 5HT2AR-targeting AAB was completely neutralized by 25 micromolar concentration of Y27632, a selective RhoA/Rho kinase inhibitor or by 50 micromolar concentration of 2-APB, an IP3R antagonist. These data are consistent with known positive coupling of TBI Ig-induced 5HT2AR and alpha 1AR signaling to Gq11/IP3R/Ca2+ and RhoA/Rho kinase signaling pathways [6].

In patients suffering with Alzheimer’s or vascular dementia, alpha 1 adrenergic receptor agonist autoantibodies were previously reported to target an epitope in the second extracellular loop of the receptor [10]. We next used enzyme linked immunoassays specific for binding to the second extracellular loop of the alpha 1 AR or the 5HT2AR to test plasma Ig from repeated and/or autoimmune TBI patients.

Alpha 1AR immunoreactivity and 5HT2AR immunoreactivity were both significantly elevated (P < 0.01) in the protein-A eluates from younger Cohort B patients who suffered repeated or autoimmune TBI (N=8), compared to level in four age-matched TBI patients lacking neurotoxicity (Figure 5A and 5B). Mean binding was approx. 2-fold higher than background (0.04 Absorbance units) in plasma harboring both alpha 1AR, and 5HT2AR AAB specificities. The Ig titer was also significantly elevated in co-occurring alpha 1AR and 5HT2AR plasmas compared to plasmas from TBI patients lacking neurotoxicity (Figure 6A-6B).

fig 5

Figure 5: Binding to a linear synthetic second extracellular loop peptide of the alpha 1A adrenergic receptor(A) or of the 5HT2A receptor (B) was significantly increased in protein-A eluates (1/40th dilution) from multiple or autoimmune TBI compared to uncomplicated TBI without bioassayable neurotoxicity. Background binding = 0.04 absorbance units. Results are mean ± SEM.

fig 6

Figure 6: Titer of binding to second extracellular loop peptide of the alpha 1A adrenergic receptor (A) or the 5HT2A receptor (B) was significantly increased in the protein-A eluates from multiple or autoimmune TBI vs. uncomplicated TBI lacking bio-assayable neurotoxicity. Background = 0.04 absorbance units.

As a further test of the specificity of recurrent TBI Ig for the alpha 1 AR and 5HT2AR, we used a synthetic peptide identical to the second extracellular loop of the alpha 2A adrenergic receptor (alpha 2R) as the target antigen in an enzyme-linked immunoassay. Since the alpha 2AR couples to Gi/Go subfamily of G proteins leading to decreased intracellular cyclic AMP, its activation is not expected to cause N2A neurite retraction. Alpha 2R agonism has peripheral sympatholytic effects (decreased blood pressure) and in the CNS mediates presynaptic inhibition of neurotransmitter release (epinephrine, norepinephrine) through actions on auto-receptors. To our knowledge, there has been no reports of spontaneously-occurring alpha 2R-targeting agonist autoantibodies in human disorders.

Mean plasma Ig binding (to the A2AR second extracellular loop peptide) in all eight younger TBI patients tested (Cohort B) was minimally elevated above background (1.25-fold) even though it was significantly higher (P < 0.05) compared to binding in four Cohort B TBI patients lacking Ig neurotoxicity (Figure  7A). The titer of A2AR peptide Ig -binding was similarly low, i.e. 1-1.25-fold vs. background (0.04 AU) in multiple TBI, autoimmune TBI, and in TBI lacking neurotoxicity (Figure 7B). Only one of twelve Cohort B patients tested (a patient with potent neurotoxicity who had suffered a single TBI) harbored alpha2R immunoreactivity at a significant level, i.e. 1.5-fold above background. The clinical significance of this finding is unknown. Taken together, these data demonstrating a much lower level of binding to the second extracellular loop of the alpha 2R, an aminergic GPCR family receptor related to the 5HT2AR [11] confirms the specificity of the results in TBI Ig for the alpha 1 AR and 5HT2AR.

fig 7

Figure 7: Absence of significant binding to a linear synthetic second extracellular loop peptide of the alpha 2A adrenergic receptor in the protein-A eluates (1/40th dilution) from multiple or autoimmune TBI compared to uncomplicated TBI lacking bio-assayable neurotoxicity. B) Non-significant, low titer of binding to alpha 2A adrenergic receptor second extracellular loop peptide in protein-A eluates from multiple or autoimmune TBI or uncomplicated TBI. Background binding = 0.04 absorbance units.

The orthosteric binding pockets of aminergic, family GPCR are conserved [9]. A novel small peptide antagonist of the 5HT2AR comprised of an epitope (SN.8) in the C-terminal region of the second ECL was previously reported to have completely neutralized the neurotoxic bioactivity in TBI and neurodegenerative disorders plasma Ig [5]. Here we tested for neutralization of N2A neurite retraction in recurrent or autoimmune TBI plasma by synthetic peptides corresponding to the entire second extracellular loop of the alpha 1A (PAPEDETICQINEEP) or the alpha 2A (RQPDAGAAYPQCGLNDET) adrenergic receptors. A twenty microgram per milliter concentration of SN.8 (SCLLADDN) completely neutralized neurotoxicity in 7.5 ug/mL concentration of autoantibodies in recurrent and single/autoimmune TBI plasma (N=4) (Figure 8A). An identical concentration of PAPEDETICQINEEP, which includes the epitope specific sequence APEDE (shown in bold text) previously reported to have blocked bioactivity in dementia plasma AAB [10] only partially prevented (52%) neurite retraction by the same four recurrent and single/autoimmune TBI plasma Igs (Figure 8B). An identical (twenty microgram per millilter) concentration of RQPDAGAAYPQCGLNDET comprising the entire second ECL of the alpha 2AR had no significant inhibitory effect on neurite retraction by two of two recurrent TBI plasma Ig tested (Figure 8C).

fig 8

Figure 8: Neurite retraction in a 1/40th dilution of the protein-A eluate from multiple or autoimmune TBI was A) nearly completely prevented by co-incubation with a 20 microgram per milliliter concentration of the SN.8 epitope-specific 5HT2AR second extracellular loop peptide, B) partially prevented by co-incubation with a 20 ug/mL concentration of a linear synthetic second extracellular loop peptide of the alpha 1A adrenergic receptor, or C) unaffected by co-incubation with a 20 ug/mL concentration of a linear synthetic peptide of the second extracellular loop of the alpha 2A adrenergic receptor. Results are mean ± SE.

A one-fourth dilution (~1 µg/mL) of the protein-G eluate of plasma in two 25-week-old male Spontaneously Hypertensive Rats (SHR) displayed two-fold increased binding to the second extracellular loop peptides of the 5HT2AR and the alpha 1A adrenergic receptors in two separate enzyme linked immunoassays (Figure 9A). The protein G eluate (1 µg/mL) from a representative twenty-five-week old male SHR caused significant N2A neurite retraction which was completely prevented by co-incubation with a twenty microgram per milliliter concentration of the SN.8 peptide (Figure 9B). Co-incubation with an identical (20 µg/mL) concentration of the PP.15 second extracellular loop peptide from the alpha1A adrenergic receptor had much less inhibitory effect on SHR Ig-induced N2A neurite retraction (Figure 9B).

fig 9

Figure 9: A) Binding to a second extracellular loop peptide of the 5HT2AR or Alpha1A AR was significantly increased in the protein-G eluate fraction of plasma in two, 25-week-old SHR rats compared to background = 0.032 AU. B) Neurite retraction in a 1/4th dilution (~1 µg/mL) of the protein-G eluate of plasma from a representative 25-week-old male SHR rat was completed prevented by co-incubation with a 20 microgram per milliliter concentration of SN..8, but was only 32% inhibited by an identical concentration of PP..15, the alpha 1A adrenergic receptor second extracellular loop peptide. Results are mean ± SEM.

Discussion

The alpha1 adrenergic receptor shares significant amino acid homology with the serotonin 2A receptor [11] in a region of the second extracellular loop involved in mediating long-lasting receptor activation. Both GPCR receptors positively couple to phospholipase C gamma/inositol triphosphate / Ca2+ release signaling pathway to promote IgG neurotoxicity in neuroblastoma cells. Repeated TBI plasma was associated with both significantly increased titer of plasma serotonin 2AR agonist IgG autoantibodies, and the appearance of additional agonist IgG having specificity for the alpha 1-adrenergic receptor based on neutralization of Ig bioactivity by a specific alpha1AR antagonist (prazosin). The striking association between multiple TBI, or single TBI/autoimmune disorder and co-occurrence of bio-assayable and immune-assayable 5HT2AR and alpha 1AR-targeting AAB suggests that specific methods for the early detection of both kinds of AAB specificities may be required to avoid missing TBI patients prone to experience rapid rate of cognitive decline. Patients with underlying systemic autoimmune disorder (e.g. autoimmune thyroid disease) who experienced only a single TBI still had the autoantibody equivalent of ‘repeated TBI exposure’ suggesting such patients may be at higher risk for future neurodegenerative complications compared to patients who experienced a single uncomplicated TBI exposure.

There are currently no medications available to slow the rate of cognitive decline in TBI patients. If future candidate drugs become available for testing, the availability of validated biomarkers (which can serve as surrogates for an increased risk of neurodegeneration) may be quite useful in identifying and monitoring high-risk TBI patients. Increased plasma 5HT2AR-immunoreactive AAB (vs lower AAB) was a significant predictor of a steep (two-year) rate of prospective cognitive decline in a cohort of older adult TBI patients from cohort A [9].

In subsets of Alzheimers’ and vascular dementia [10] autoantibodies targeted an epitope in the second ECL of the alpha 1AR (shown in bold, PAPEDETICQINEEP) located in a region N-terminal to the conserved (underlined) cysteine residue. On the other hand, 5HT2AR- targeting autoantibodies in diverse neurodegenerative pathologies were reported to target (QDDSKVFKEGSCLLADDN) a more highly conserved region of the second extracellular loop (shown in bold letters) including (underlined) amino acid residues which play a key role in mediating aminergic receptor activation [11]. For example, conserved amino acid residues leucine (L) and aspartate (D) play key roles in stabilizing hydrophobic interactions within the transmembrane core (L) and interhelical hydrogen bonding (D) important in activating GPCR signaling, respectively. Since the small hydrophobic SN.8 peptide (SCLLADDN) antagonist contains these two conserved amino acid residues it might ‘compete’ with the same amino acid residues on the native receptor for binding sites important in stabilizing and activating the receptor.

Persistent neuro-inflammation associated with repeated TBI and single/autoimmune TBI may increase the likelihood for occurrence of agonist autoantibodies directed against closely-related catecholamine receptors. Catecholamine receptors are expressed on immune cells and play a role in promoting acute and chronic inflammatory responses [12,13]. Treatment with either 5HT2AR [14] or alpha 1 AR antagonist medication [15] was previously reported to be associated with reduced mortality in severe Covid 19 infection. Plasma agonist 5HT2AR IgG autoantibodies were associated with hyperinflammation in severe Covid-19 infection and increased titer was associated with an increased risk of Covid-19 mortality [16].

Alpha 1 adrenergic receptor autoantibodies were previously reported to increase in subsets of human refractory hypertension [12], however, in the present study only two of forty-two patients (~5%) suffered with refractory hypertension. Alteration in regulatory T cells has been linked with hypertension in the genetically hypertensive male Spontaneously Hypertensive Rat (SHR) strain [17]. Male SHR rats (25-weeks-old) not subjected to TBI, spontaneously harbored both alpha 1A adrenergic and 5HT2A receptor immunoreactive autoantibodies, and the level and titer of the two kinds of plasma catecholaminergic agonist AAB were nearly indistinguishable. Acute N2A neurite retraction induced by male SHR Ig was not only prevented by the 5HT2A second extracellular loop receptor peptide SN.8 (in vitro), but systemic (in vivo) administration of SN.8 in male SHR rats was reported to mediate potent, long-lasting systolic and diastolic blood pressure-lowering [7].

Alpha 1 AR and 5HT2AR are expressed in overlapping, but distinct brain regions. Decreased expression of each receptor was reported in cerebral cortex from dementia patients [18,19] perhaps consistent with shared downstream Gq11/IP3/Ca2+ signaling pathway activation (by AAB) which is toxic in neurons. There has been no prior report of a specific association between alpha 1 AR agonist autoantibodies and TBI. A much larger study is needed to test the hypothesis that alpha 1AR autoantibodies may increase in association with specific symptoms in TBI with chronic PTSD (e.g. trauma nightmares, panic disorder, anxiety).

In summary, plasma alpha 1 adrenergic and serotonin 2A receptor agonist AAB co-occur in patients who experienced repeated TBI or single TBI in the setting of systemic autoimmunity and may contribute to higher risk for dementia. A small peptide antagonist, SN.8 comprised of a highly conserved region of the serotonin 2A receptor whose amino acid residues play key roles in stabilizing aminergic GPCR activation, prevented neurotoxicity from both 5HT2AR- and alpha 1AR-targeting autoantibodies by an unknown mechanism. These observations may have particular relevance to patients expressing both kinds of autoantibodies (without a history of TBI exposure) as the results of a recent study in the Zucker rat strain (which expressed both 5HT2AR and alpha 1AR AAB) indicated significant in vivo neuroprotection (by systemically-administered SN.8) in sham-injured, but not in TBI-injured rats [20].

Acknowledgements

Supported by a grant from the New Jersey Commission of Brain Injury Research (Trenton, New Jersey) NJCBIR22 PIL022 to MBZ.

References

  1. Walker KR, Tesco G (2013) Molecular mechanisms of cognitive dysfunction following traumatic brain injury. Frontiers in Aging Neuroscience 5: 29. [crossref]
  2. Charney DS, Woods SW, Nagy LM, Southwick SM, Krystal JH, et al. (1990) Noradrenergic function in panic disorder. J Clin Psychiatry 51: 5-11. [crossref]
  3. Raskind MA, Peskind ER, Hoff DJ, Hart KL, Holmes HA, et al. (2007) A parallel group placebo- controlled study of prazosin for trauma nightmares and sleep disturbance in combat veterans with post-traumatic stress disorder. Biol Psychiatry 61: 928-934. [crossref]
  4. Zimering MB, Pulikeyil AT, Myers CE, Pang KC (2020) Serotonin 2A Receptor Autoantibodies Increase in Adult Traumatic Brain Injury in Association with Neurodegeneration. J Endocrinol Diabetes 7: 1-8. [crossref]
  5. Zimering MB (2019) Autoantibodies in Type-2 Diabetes having Neurovascular Complications Bind to the Second Extracellular Loop of the 5-Hydroxytryptamine 2A Receptor. Endocrinol Diabetes Metab J 3: 118. [crossref]
  6. Zimering MB (2017) Diabetes Autoantibodies Mediate Neural- and Endothelial Cell- Inhibitory Effects Via 5-Hydroxytryptamine- 2 Receptor Coupled to Phospholipase C/Inositol Triphosphate/Ca2+ Pathway. J Endocrinol Diabetes 4: 10. [crossref]
  7. Grinberg M, Zimering MB (2022) Myristolated Serotonin 2A Receptor Peptide Promotes Long-Lasting Blood Pressure-Lowering and Reno protection in Hypertensive Rat Species. Endocrinol Diabetes Metab J 6. [crossref]
  8. Zimering MB, Grinberg M, Burton J, Pang K (2020) Circulating Agonist Autoantibody to 5-Hydroxytryptamine 2A Receptor in Lean and Diabetic Fatty Zucker Rat Strains. Endocrinol Diabetes Metab J 4: 413. [crossref]
  9. Zimering MB, Grinberg M, Myers CE, Bahn G (2022) Plasma Serotonin 2A Receptor Autoantibodies Predict Rapid, Substantial Decline in Neurocognitive Performance in Older Adult Veterans with TBI. Endocrinol Diabetes Metab J 6: 614. [crossref]
  10. Wallukat G, Prüss H, Müller J, Schimke I (2018) Functional autoantibodies in patients with different forms of dementia. PLoS One 13: e0192778. [crossref]
  11. Michino M, Beuming T, Donthamsetti P, Newman AH, Javitch JA, et al. (2015) What can crystal structures of aminergic receptors tell us about designing subtype-selective ligands?. Pharmacol Rev 67: 198-213. [crossref]
  12. Wenzel K, Haase H, Wallukat G, Derer W, Bartel S, et al. (2008) Potential relevance of alpha(1)-adrenergic receptor autoantibodies in refractory hypertension. PLoS One 3: e3742. [crossref]
  13. Grisanti LA, Perez DM, Porter JE (2011) Modulation of immune cell function by α(1)-adrenergic receptor activation. Curr Top Membr 67: 113-138. [crossref]
  14. Zimering MB, Razzaki T, Tsang T, Shin JJ (2020) Inverse Association between Serotonin 2A Receptor Antagonist Medication Use and Mortality in Severe COVID-19 Infection. Endocrinol Diabetes Metab J 4: 1-5. [crossref]
  15. Rose L, Graham L, Koenecke A, Powell M, Xiong R, et al. (2021) The Association between Alpha-1 Adrenergic Receptor Antagonists and In-Hospital Mortality From COVID-19. Front Med Lausanne 8: 637-647. [crossref]
  16. Zimering MB (2021) Severe COVID-19 Pneumonia is Associated with Increased Plasma Immunoglobulin G Agonist Autoantibodies Targeting the 5-Hydroxytryptamine 2A Receptor. Endocrinol Diabetes Metab J 5: 1-9. [crossref]
  17. Katsuki M, Hirooka Y, Kishi T, Sunagawa K (2015) Decreased proportion of Foxp3+ CD4+ regulatory T cells contributes to the development of hypertension in genetically hypertensive rats. J Hypertens 33: 773-783. [crossref]
  18. Lai MK, Tsang SW, Alder JT, Keene J, Hope T, et al. (2005) Loss of serotonin 5-HT2A receptors in the postmortem temporal cortex correlates with rate of cognitive decline in Alzheimer’s disease. Psychopharmacology 179: 673-677. [crossref]
  19. Szot P, White SS, Greenup JL, Leverenz JB, Peskind ER, et al. (2007) Changes in adrenoreceptors in the prefrontal cortex of subjects with dementia: evidence of compensatory changes. Neuroscience 146: 471-480. [crossref]
  20. Grinberg M, Burton J, Pang KC, Zimering MB (2023) Neuroprotective Effects of a Serotonin Receptor Peptide Following Sham vs. Mild Traumatic Brain Injury in the Zucker Rat. org 2023, 2023050004.