Medicine
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This repository contains the published and unpublished research of the Faculty of Medicine by the staff members of the faculty
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Item Significance of evaluating immature germ cells during semen analysis(Jaypee Brothers Medical Publishers, 2022) Dissanayake, D.M.A.B.Item Differentiation of human male germ cells from Wharton's jelly-derived mesenchymal stem cells(Korean Society for Reproductive Medicine, 2018) Dissanayake, D.M.A.B.; Patel, H.; Wijesinghe, P.S.OBJECTIVE: Recapitulation of the spermatogenesis process in vitro is a tool for studying the biology of germ cells, and may lead to promising therapeutic strategies in the future. In this study, we attempted to transdifferentiate Wharton's jelly-derived mesenchymal stem cells (WJ-MSCs) into male germ cells using all-trans retinoic acid and Sertoli cell-conditioned medium. METHODS: Human WJ-MSCs were propagated by the explant culture method, and cells at the second passage were induced with differentiation medium containing all-trans retinoic acid for 2 weeks. Putative germ cells were cultured with Sertoli cell-conditioned medium at 36℃ for 3 more weeks. RESULTS: The gene expression profile was consistent with the stage-specific development of germ cells. The expression of Oct4 and Plzf(early germ cell markers) was diminished, while Stra8 (a premeiotic marker), Scp3 (a meiotic marker), and Acr and Prm1 (postmeiotic markers) were upregulated during the induction period. In morphological studies, approximately 5% of the cells were secondary spermatocytes that had completed two stages of acrosome formation (the Golgi phase and the cap phase). A few spermatid-like cells that had undergone the initial stage of tail formation were also noted. CONCLUSION: Human WJ-MSCs can be transdifferentiated into more advanced stages of germ cells by a simple two-step induction protocol using retinoic acid and Sertoli cell-conditioned medium.Item Sexual practices, knowledge and usage of contraceptives among adolescents and young adults(Sri Lanka College of Obstetricians and Gynaecologists, 2008) Herath, H.M.R.P.; Dissanayake, D.M.A.B.; Hilmi, M.A.M.; Pathmeswaran, S.; Fernando, W.S.; Wijesinghe, P.S.INTRODUCTION: Adolescence is a critical period of development, as important behaviour patterns are learnt during this period. The sexual debut in Sri Lankan schooling adolescents has been reported as 15.3 years for males and 14.4 for females. Furthermore a significant difference in the prevalence of heterosexual experience among the two sexes have been reported(14% among boys Vs 2% among girls). In 2001, of the pregnant women who received antenatal care from public health midwives, 7.8% were teenagers. Interestingly 22.2% of maternal deaths due to illegal abortions were in this age group during 1996. Thus the situation demands research into sexual practices and strategies to increase the contraceptive uptake by teenagers. OBJECTIVES: To describe and compare the sexual practices. knowledge and usage of contraceptives by the adolescents and young adults with different educational backgrounds. METHOD: Study was a cross sectional descriptive study done involving subjects of three different educational levels i.e. students of universities and technical colleges and employees of garment factories. Data were collected with a pretested self administered questionnaire. RESULTS: Data of 1258 subjects were analyzed. Of them 23% were garment factory workers, 38.2% were technical college students and 38.8% were university students. Females consisted 58.9% of the population. The mean age for males and females were 20.5 years and 20.7 years respectively. The mean age of first sexual relationship of males and females were at 17.65 and 19.44 years respectively (p<.001). Out of the total population 17% of females and 33.5% of males were sexually active. Sexual relationships were reported by 51.7% of garment factory employees, 19.4% technical college students and 12.3% of university students. Majority of male garment factory workers ("90 %) were sexually active. Of the 166 sexually active males 19.3% had sexual relationship with commercial sex workers. Out of the 172 people who have not used modern contraceptives during heterosexual relationships, 32% has used natural family planning. Significantly larger proportion (21.7%) of garment factory workers compared to other settings believed that IUCD would protect them from HIV infection. Of the study population 24.9% of females and 71.2% of males knew barrier methods would minimize the risk of development of HIV. With regard to primary source of information regarding contraceptive methods, 24.2 % and 23.4% of subjects reported school and news papers, while 28.4% felt that information from parents as the least important source. Of the people who selected school as their main source of information, only 41.4% knew barrier methods could protect them from HIV. Majority preferred instructions from school and special health clinics on contraceptive usage. CONCLUSIONS: Teenage and young adult males have their first sexual relationship at an earlier age than female counterparts. A larger proportion of garment factory employees are sexually active compared to students of technical colleges and universities. Significant proportion of males of the age group had their first relationship with commercial sex workers. The knowledge about different contraceptive methods among this population is inadequate. More emphasis should be given to the preferred media by the adolescents and young adults in educating them on contraceptive methods.Item Semen ph in infertile males and its association with semen parameters(Sri Lanka College of Obstetricians and Gynaecologists, 2005) Dissanayake, D.M.A.B.; Palihawadana, T.S.; Amarasena, J.M.T.; Fernando, W.S.INTRODUCTION: Assessment of pH of the ejaculate is part of the basic seminal fluid analysis. The WHO defines the reference range for seminal fluid pH to be 7.2 or more. In practise we have observed that the pH is commonly in the alkaline range in patients with normal as well as abnormal seminal parameters. Furthermore, there are many others who question the reference range defined by the WHO, and claim it to be too low. Objective: To define the range of pH in seminal fluid in a infertile male population and to assess the association it shows to seminal fluid parameters. METHOD: Seminal fluid analysis including pH assessment was done on 168 males who sought infertility treatment at the university infertility clinic, at teaching hospital, Ragama. The pH was assessed within 30 minutes from collecting the sample using a pH meter. The seminal fluid analysis was done according to the WHO criteria. Log inversion of pH was done prior to calculation ofmeans. Results: The mean (±SD) pH of the sample population was 7.70 (±0.25) with a minimum of 6.78 and a maximum of 8.52. Only five patients (3%) had a pH less than 7.2. The distribution of the pH showed a 5" percentile value of 7.37 and a 95 'percentile of8.21.Thirty four percent (n=57) of subjects had a normal Seminal fluid analysis with regard to volume, concentration, motility and viability. Mean pH value showed a significant difference between the groups with a normal vs. abnormal volume of ejaculate, 7.75 vs. 7.59, P<0.05. Similar differences were not seen with any other semen parameter. Conclusion: The pH of ejaculate in this study sample is towards an alkaline range. Associations between semen pH and seminal parameters needs further study.Item In vitro spermatogenesis: a myth or reality?(Faculty of Medicine, University of Kelaniya, Sri Lanka, 2016) Dissanayake, D.M.A.B.Spermatogenesis is a complex, unique and tightly coordinated process takes place in the seminiferous tubules. It is divided broadly into three distinct phases; proliferation of spermatogonial stem cells (mitosis), reduction of chromosome number (meiosis) and transformation of spermatids into sperm (spermiogenesis). Recapitulation of this orchestrated process in vitro is significant in understanding germ cell biology, onset of some inherited disorders, treatment of infertility, and deciding methods of genetic manipulations and fertility preservation. Different attempts have been made using embryonic stem cells, somatic stem cells, induced pluripotent stem cells and spermatogonial stem cells to achieve this goal. Studies have shown the potential capacity of stem cells to differentiate into germ cells under appropriate culture conditions. However, producing a functionally normal sperm is still a greater challenge for scientists. Results of our recent study indicated that umbilical cord blood derived mesenchymal stem cells can differentiate in to male germ cells lineage when induction with retinoic acid and co-culture with Sertoli cells. However, we could reach only up to post meiotic spermatid level as shown by majority of studies. Lack of epididymal support and correct endocrine profile may be the reason behind this failure. We have planned to elucidate this hypothesis in a future research.Item Adolescent sexual practices and contraceptive usage(Faculty of Medicine, University of Kelaniya & Plan International, 2008) Herath, H.M.R.P.; Dissanayake, D.M.A.B.; Hilmi, M.A.M.; Pathmeswaran, A.; Wijesinghe, P.S.INTRODUCTION: Adolescence is a critical period of development, as adoption of unhealthy risk behaviours such as unprotected sex, smoking and drug abuse, avoiding contraceptive methods will lead to long standing health and socioeconomic consequences. Therefore information is needed about the sexual practices and contraceptive usage of adolescents and young adults to develop interventions in the community. This study was undertaken to describe and compare the sexual practices, knowledge and usage of contraceptives by adolescents and young adults in selected urban and rural settings.METHOD: This cross sectional descriptive study was conducted among young adults aged less than 21 years at the time of interview in three settings in both rural and urban areas. The study subjects were students from universities, technical colleges and garment factory workers representing both rural and urban areas. The data was collected using a pretested self administered questionnaire during June 2007 to August 2007. Information collected included marital status, gender and the level of education, first sexual relationship (age, partner, contraception used), current contraceptive practices and knowledge about contraceptive methods. The data was analysed using SPSS 10.1 statistical package. Informed consent was obtained from all participants. RESULTS : There were a total of 1258 subjects who had returned the completed questionnaires. Out of them there were 290 garment factory employees, 480 technical college students, and 488 university students. The percentage of females was 58.9%. The mean ages of the males and the females of the total population were 20.75(50 1.13) years and 20.55(SD 1.63) and it was similar in the three settings. The level of education was highest among the undergraduates lowest among garment factory workers. Out of the population 303 (24.1%) were sexually active. It was significantly higher among males {33.5%0 than females (17.0%). Both among male and females this trend was significantly more common among garment factory workers. Sexual activity of the unmarried and the premarital sexual practices of married subjects are shown in the table. 6. 17% of females and 33.5% of males were sexually active before marriage. 90% male garment factory workers were sexually active. In contrast the proportionItem A preliminary study on bacteriospermia in males of subfertile couples(Sri Lanka Medical Association, 2011) Abeysundara, I.D.H.P.K.; Dissanayake, D.M.A.B.; Wijesinghe, P.S.; Perera, R.R.D.P.INTRODUCTION AND OBJECTIVES: Bacteriospermia is common in subfertile men. Responsible organisms and their antibiotic sensitivity patterns vary between populations Aim of this study was to identify the types of bacteria and their ABST pattern among males of subfertile couples. METHODS: A descriptive cross sectional study was carried out. Semen samples of 105 males from subfertile couples were cultured in blood, chocolate and McConkey agar. Antibiotic sensitivity tests were done using the Clinical and Laboratory Standard Institute technique. RESULTS: Mean age of the study population was 32.77 yrs (rage 22-48 yrs), mean duration of subfertility was 3 yrs (Range 1-12 yrs). In 98 couples there was primary subfertility. Bacterial growth was noted in 63 samples. Positive cultures were found in 58.2% with primary subfertility and in 85.7% with secondary subfertility. A single organism was cultured in 44. Mltiple organisms were cultured in 19 samples. In 21 samples a significant growth of >104 of colony forming units were noted. Organisms found were Streptococci (n=28 out of them 8 were group D), coagulase negative Staphylococci (n=27), Diptheroids (n=T2), coliforms (n=7 of which 3 were Escherichia colt), Staphyhcoccus aureus (n^5), Acinetobacter (n=2), and Candida albicans (n=l). A single antibiotic or a combination which covers most organisms was not found. CONCLUSIONS: In this study population 20% had a significant bacterial growth. Organisms were found to have varied ABST patterns. The effect of bacteriospermia on subfertility needs further evaluation. Acknowledgements: This study was supported by National Research Council (09-69).Item A Study on Factors Associated with Clomifene Resistance in Infertile Women with WHO Group II Anovulation at a Teaching Hospital in Sri Lanka(Sri Lanka College of Obstetricians and Gynaecologists, 2014) Palihawadana, T.S.; Dissanayake, D.M.A.B.; Wijesinghe, P.S.; Seneviratne, H.R.Background: Anovulation is a common cause of infertility and induction of ovulation with oral agents such as clomifene is the treatment of choice in women with normogonadotropic anovulation. While such treatment is able to achieve ovulation in many, some women fail to respond thus termed as having clomifene resistance. Knowledge on factors associated with such resistance would enable us to counsel patients more accurately and offer alternative treatment modalities without undue delays. This study was aimed at identifying factors associated with clomifene resistance among infertile women with WHO group II anovulation at a teaching hospital in Sri Lanka. Method: A case control study was done at the Infertility clinic of the North Colombo teaching hospital, Ragama conducted by the department of obstetrics & Gynaecology of the University of Kelaniya. Women with WHO group II anovulation (n=128) were included and underwent induction of ovulation with clomifene citrate starting at a dose of 50mg per day, increased up to 150mg till ovulation was achieved. Those who fail to respond to a maximum dose of 150mg were termed as clomifene resistant. The factors known to be associated with clomifene resistance were compared between the two groups who responded and those who failed to respond. The study was registered with the Sri Lankan clinical trial registry maintained by the Sri Lanka Medical association. The study was funded by the National Science foundation of Sri Lanka through a research grant (Grant No: RG/2007/HS/08) Results: Ovulation was achieved in 77.3% (n=99) of subjects and therefore the prevalence of clomifene resistance was 22.8% (n=29). Among those who responded, 41 did so at a dose of 50mg per day while 40% did with 100mg, thus over 80% achieving ovulation at a dose of 100mg per day or less. The factors associated with clomifene resistance in this population included infertility of more than three years (OR 2.06, 95%CI 1.51-9.14), presence of hirsutism (OR 2.76, 95%CI 1.18-6.46), a higher AFC (16 vs 13, p=0.04), PCOS (OR 2.99, 95%CI 1.06-8.41) and an LH:FSH ratio of > 1(OR3.11, 95%CI 1.33-7.24). Other factors that have been described to be associated with clomifene resistance such as advanced age, a BMI > 25, and a higher ovarian volume did not show any significant associations in this study. Discussion: This study was able to identify factors associated with clomifene resistance in a local population. Most factors identified included clinical and laboratory findings of PCOS thus highlighting the limitations in treatment success of clomifene among women with PCOS. This information may be used in clinical practice to counsel patients prior to treatment or to consider other alternative treatment options such as Letrozole or gonadotropins. DOI: http://dx.doi.org/10.4038/sljog.v36i3.7715Item Zinc levels in seminal plasma and its relationship with semen parameters in male partners of subfertile couples(Sri Lanka Association for the Advancement of Science, 2006) Dissanayake, D.M.A.B.Item Evaluation of reproductive hormone profile in relation to semen quality in male partners of subfertile couples(Sri Lanka Association for the Advancement of Science, 2007) Dissanayake, D.M.A.B.; Palihawadana, T.S.; Wijesinghe, P.S.; Ratnasooriya, W.D.; Wimalasena, S.The objective of the study was to assess the reproductive hormone profile of a Sri Lankan subfertile male population and to relate it to their semen parameters. Male partners of 152 subfertile couples who sought treatment at the subfertility unit of the department of Obstetrics and Gynaecology at the Faculty of Medicine, Ragama were included in the study. Serum samples were assayed for follicle stimulating hormone (FSH), luteinizing hormone (LH), prolactin (PRL), Estradiol (E2) and testosterone (T) levels, using electrochemiluminescence immunoassay (ECLIA). A semen sample was obtained from each participant on the same day. Seminal fluid analysis was done according to the WHO guidelines. The mean (SEM) of FSH, LH, PRL, E2 and T levels of the males were 5.35 (0.53) mIU/mL, 6.20 (0.33) mIU/mL, 17.53 (0.60) ng/mL, 30.48 (1.80) pg/mL and 523.80 (14.74) ng/dL and the prevalence of abnormalities of these hormones were 9.8% (n=15), 5.9% (n=9), 13.15% (n=20), 0.6% (n=1) and 2.6% (n=4) respectively. Of the study population 20.3% (n=31) men had either single or combination of hormone abnormalities. When the mean hormone levels of the men with normal semen parameters were compared with those of men with abnormal semen parameters, none showed a statistically significant difference; FSH - 4.18 (2. 78) Vs 4.58 (3.15), LH - 5.90 (2.78) Vs 5.70 (2.36), PRL - 28.63 (14.96) Vs 17.59 (7.63) and T -551.39ñ208.17 Vs 515.73 (158.25). Mean (SD), FSH and LH levels were significantly higher among azoospermic participants compared with normozoospermics; 19.69 (9.93) Vs 4.18 (2.78) for FSH, 12.82 (11.82) Vs 5.90 (2.78) for LH, p<0.01). Similarly FSH and LH levels were significantly higher in severe oligospermic participants compared with the normozoospermics; 7.58 (4.34) Vs 4.18 (2.78) for FSH, and 6.96 (2.65) Vs 5.90 (2.78) for LH, p<0.05). The serum FSH level showed an inverse correlation with sperm concentration (r= -0.203, p<0.05) and total sperm count (r= -0.206, p<0.05). In conclusion, an endocrinopathy was present in up to 20% of this population. Abnormalities in Gonadotrophin levels were related with azoospermia and severe oligozoospermia. Serum FSH showed a negative correlation with the sperm concentration and the total sperm count. Other hormones failed to show a significant relationship with the semen parameters in this population. Acknowledgement: National Science Foundation research grant No. RG/2004/M/14