Dissertations / Theses on the topic 'Rats Infertility'

Orientador: Wilma de Grava Kempinas
Dissertação (mestrado) - Universidade Estadual de Campinas, Instituto de Biologia
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Resumo: O Diabetes mellitus geralmente se associa a algum tipo de disfunção sexual, provocando infertilidade, tanto em seres humanos, quanto em animais experimentais. Em trabalho anterior realizado em nosso laboratório, ratos machos que tiveram hiperglicemia induzida quimicamente pela administração de streptozotocin apresentaram diminuição da fertilidade através de múltiplos parâmetros analisados. O presente estudo teve como objetivo investigar os mecanismos envolvidos e o papel da testosterona no processo. Para tanto, ratos machos foram divididos em 3 grupos experimentais: normoglicêmico (controle), hiperglicêmico (streptozotocin), e hiperglicêmico com reposição hormonal (streptozotocin+testosterona) e foram avaliados parâmetros reprodutivos e espermáticos, níveis hormonais, a contratilidade do ducto deferente isolado in vitro, comportamento sexual e o número de espermatozóides ejaculados no útero. O ducto deferente de animais diabéticos apresentou um quadro de hipersensibilidade à metoxamina, um agonista sintético de adrenoceptores _1. Estes mesmos animais apresentaram os seguintes resultados: alteração de comportamento sexual e ausência de espermatozóides ejaculados, redução dos níveis plasmáticos de testosterona, perda de peso corpóreo e de órgãos como epidídimo, ducto deferente, vesículas seminais e próstata ventral, perda de células germinativas na luz e desorganização epitelial aparente em túbulos seminíferos, além da aceleração do tempo de trânsito dos espermatozóides pelo epidídimo. Os dados apresentados neste trabalho indicam que os mecanismos responsáveis pela perda de fertilidade natural de ratos diabéticos envolvem comprometimento do processo espermatogênico, assim como desregulação do eixo reprodutivo masculino, juntamente com evidências para problemas no processo de maturação espermática, tendo como fator complicante o prejuízo da função ejaculatória, dependente da contratilidade da musculatura lisa dos ductos deferentes. A reposição de andrógeno não foi totalmente capaz de reverter os danos causados pelo diabetes no sistema reprodutivo masculino de ratos adultos
Abstract: Diabetes mellitus is usually related with some kind of sexual dysfunction, promoting infertility in humans as well as in experimental models. In a prior work from our laboratory, male rats, which had a diabetic-induced state of hyperglycemia caused by streptozotocin administration, demonstrated reduced fertility through several parameters analyzed. The present study aimed at investigating the mechanisms involved and the role of testosterone in the process. Male rats were randomly allocated in 3 experimental groups: control, hyperglycemic (streptozotocin), and hyperglycemic with hormone replacement (streptozotocin+testosterone) and the following parameters were analyzed: reproductive and spermatic parameters, hormone levels, sexual behavior, contractility of vas deferens in vitro, sexual behavior parameters and the number of sperm ejaculated in utero. The vas deferens of diabetic animals was hypersensitive to methoxamine, a synthetic agonist of _1 adrenoceptors. The same animals showed the following results: alterations in sexual behavior and lack of sperm ejaculated, reduction in plasma testosterone levels, decreased body weight and epididymis, seminal vesicles, ventral prostate and vas deferens weights, loss of germ cells in the lumen and apparent epithelial disarrange in seminiferous tubules, and accelerated sperm transit time in the epididymis. The data presented herein indicate that the mechanisms underlying the reduced fertility through natural mating observed in diabetic rats involve impairment of the spermatogenic process, as well as a dysregulation of the male reproductive axis, together with evidence for problems in the sperm maturation process, which has as a complicant factor the impairment of the ejaculatory function, dependent on the vas deferens smooth muscle contractility. Androgen replacement was not totally capable of reversing the damage caused by diabetes on the male reproductive system of adult rats
Mestrado
Biologia Celular
Mestre em Biologia Celular e Estrutural

Infertility is one of the major problems of obesity. Studies have shown that administration of leptin reversed obesity-induced infertility in rats and mice. Coenzyme Q10 (CoQ10) is an antioxidant and also supplies the energy needed for ovulation and embryo development. We hypothesized that leptin when combined with CoQ10 could greatly improve obesity-induced infertility. The results showed a significant decrease in food intake, body weight, and the regular estrous cycle was restored after treatment with leptin+CoQ10. There was a significant increase (p10 significantly (p10 can improve fertility in obese infertile female rats. This study could provide a novel therapeutic strategy for the treatment of infertility and formulation of new drugs for the treatment of obesity-induced infertility in females.

Infertility is one of the major complications of obesity. Studies have shown that administration of leptin modulated the expression of Β-catenin in the ovary and reversed obesity-induced infertility in rats. Coenzyme Q10 (CoQ10), an antioxidant, supplies the energy used for ovulation, oocyte and embryo development and prevents DNA damage that causes infertility. We hypothesized that leptin when combined with CoQ10 could greatly improve fertility. Twenty-one female Sprague-Dawley rats were used in this study and divided into five treatments groups. Group I rats was fed rat chow diet (RCD) while groups II to V were fed High-fat diet (HFD) for 14 weeks to induce infertility. Group 1 RCD and group II HFD control rats received 1 ml of saline intraperitoneally (i.p.) twice daily for 2 days, group III HFD rats received 1 ml of 100 µg of leptin i.p. twice daily for 2 days, group IV HFD rats received 10 mg/kg of CoQ10 i.p. for 2 weeks plus saline twice daily for 2 days. Group V HFD rats received 1 ml of 100 µg of leptin i.p. twice daily for 2 days plus 10 mg/kg of CoQ1o i.p. for 2 weeks. Estrous cycle was checked daily and food intake and body weight measured twice weekly before and after treatments. Fourteen days post treatment, all the animals were sacrificed. The blood and tissues were collected for analysis. The results show a significant decrease in food intake and body weight and regular estrous cycle restored in groups III and V rats. There was significant (p < 0.05) increase in spleen weight in groups IV and V. FSH level increased significantly (p < 0.05) in the leptin plus CoQ10 treated group while CoQ10 level was increased significantly (p < 0.05) in the leptin-treated group. Β-catenin expression was decreased in group IV and V, suggesting that Β-catenin expression may be downregulated by COQ10 administration. These results indicate that synergistic action of leptin and CoQ10 could delay the onset of obesity-induced infertility exhibited by the reduction of food intake and body weight. In conclusion, combinations of CoQ10 with leptin can improve fertility in obese infertile female rats and could provide a novel therapeutic strategy for the treatment of female infertility.

One of the numerous complications of obesity is infertility. Leptin has been shown to reverse infertility; however, exact mechanism is poorly understood. Recent evidence indicates Ecadherin/ β-catenin complex, which is a structural constituent of adherens junction, is expressed in the rat ovary during folliculogenesis. We hypothesized that systemic leptin modulates the expression of E-cadherin and β-catenin in dietary-induced obese infertile rats to reverse infertility. Female Sprague-Dawley rats were fed either regular chow diet (RCD) (n=6) or high fat diet (HFD) (n=14). Oestrus cycles were monitored daily until their cycles became irregular. 100 ug/ml of leptin was given intraperitoneally to HFD-fed rats (n=5) with irregular cycles. The control rats HFD (n=9) and RCD received saline. Leptin treatment restored regular estrous cycle and increased the expression of E-cadherin and β-catenin in all the 5 rats (HFD+Leptin). This could represent the mechanism by which leptin reverses infertility in obese infertile rats.

Thesis (MTech (Biomedical Technology))--Cape Peninsula University of Technology, 2015.
Diabetes mellitus (DM) has been reported to be one of the greatest global public health threats. Statistics of the fertility status of modern society has linked increased DM to a decrease in fertility rates. Hyperglycaemia is characteristic of DM that results in a disturbance of proteins, lipids and carbohydrate metabolism leading to an increase production of reactive oxygen species (ROS). In the case where ROS overwhelms antioxidant mechanisms, the body goes into state of oxidative stress (OS). OS plays a vital role in the progression of DM which leads to dysfunction and damage of various organs including that of the reproductive system. Os has shown to cause damage to the sperm membraneby oxidation of polyunsaturated fatty acids (PUFA’s) as the sperm membrane are rich in PUFA’s. This damage contributes to reduced sperm motility, concentration, morphological abnormalities and the sperms ability to fuse with the ZP of the oocyte. DM has been observed to cause testicular degeneration by interrupting sertoli cell production and maintenance thus resulting in a disturbance of the normal functioning of the reproductive system. Experimental studies have targeted more natural sources for treating DM and its complications of the reproductive system. Plants and natural dietary substances have shown to have high antioxidant contents that combat DM induced oxidative stress. This study explored the effect the Hypoxis hemerocallidea (H. hemerocallidea) supplementation on testicular and epididymal tissue, sperm motility and reproductive hormones in male wistar rats. The experiment were conducted for 6 weeks and the rats (230-260 grams) were randomly divided into 5 groups (n=12 per group). Diabetes was induced in 3 of the 5 groups. The first group was the normal control group (A), second the diabetic control group (B), third was the diabetic group treated with 800mg/kg H. hemerocallidea (group C), fourth the diabetic group treated with 200mg/kg H. hemerocallidea (group D) and fifth the non-diabetic group supplemented with 800mg/kg H. hemerocallidea (group E). Blood glucose showed a significant increase in the diabetic group when compared to the normal control and treated groups. H. hemerocallidea showed improvement in sperm motility and sperm morphology more at 800mg/kg when compared to diabetic group and diabetic group treated with 200mg/kg. Body, testicular and epipidymal weights of diabetic control were significantly lower when compared to the other groups. Testicular and epididymal Malondialdehyde levels were decreased in normal control, diabetic groups treated with different doses of H. hemerocallidea and the non-diabetic group supplemented with H. hemerocallideaon comparing with the diabetic control group. Antioxidants such as Superoxide dismutase, Catalase and total Glutathione activity was observed to be dosage dependent in certin groups but most showed a significant increase when compared to the diabetic control group. The total antioxidant capacity was measured using Oxygen radical absorbance capacity (ORAC) and Ferric ion reducing antioxidant power (FRAP); increase was observed when normal control group and treated groups were compared to the diabetic group. Testosterone and estradiol levels were also increased when the normal control group and treated groups were compared to the diabetic control group. Based on our findings it can be concluded that H. hemerocallidea supplementation can potentially be used to counteract deleterious effects of DM on the male reproductive system.

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FAPEMIG - Fundação de Amparo à Pesquisa do Estado de Minas Gerais
Os dados estatísticos de prevalência e incidência de sobrepeso/obesidade vem aumetando ao longo dos anos e atinge milhões de pessoas ao redor do mundo, tornando-se assim, umas das grandes epidemias do século XXI. A globalização e o aumento na disponibilidade de alimentos mais palatáveis e ricos em gordura saturada, somados ao estilo de vida sedentário, contribuem diretamente no desenvolvimento dessa condição. Estudos demonstram que alterações nutricionais durante fases críticas do desenvolvimento infantil, como a lactação, estão associadas ao desenvolvimento de obesidade, alterações metabólicas, doenças cardiovasculares, problemas respiratórios e diabetes. Além disso, o sobrepeso/obesidade pode influenciar na função reprodutiva da população, gerando malformações fetais, infertilidade, assim como morte materna e fetal. Considerando o exercício físico como fator primário no combate ao sobrepeso/obesidade, pretendeu-se avaliar se o exercício físico, em diferentes intensidades, é capaz de influenciar na saúde reprodutiva de indivíduos com sobrepeso/obesos. Foram utilizadas 40 ratas Wistar provenientes de ninhadas padrão (NP=10 filhotes/mãe) e de ninhadas reduzidas (NR=4 filhotes/mãe) que promoveram o sobrepeso/obesidade. Os animais foram treinados durante oito semanas em protocolos de intensidade moderada (Endurance) e intervalar de alta intensidade (Hiit), acasalados aos 90 dias de vida e eutanasiados no 20o dia de gestação. As variáveis metabólicas, bioquímicas e reprodutivas foram coletadas e analisadas. Os animais dos grupos superalimentados treinados Endurance e Hiit apresentaram redução significativa das médias de peso, consumo, índice de tolerância à insulina e de determinados parâmetros bioquímicos e reprodutivos analisados quando comparados com o grupo superalimentado sedentário. Porém, não houve diferenças significativas com relação ao comprimento nasoanal; peso relativos de órgãos; gorduras retroperitoneal e perigonadal; além do teste de tolerância oral à glicose; índices de gestação, implantação e reabsorção fetal; e demais análises fetais nos grupos estudados. Concluiu-se que o treinamento físico, em suas diferentes intensidades, não foi capaz de melhorar significativemente a capacidade reprodutiva desses animais, entretanto, foi eficiente em promover redução do peso, consumo, sensibilidade à insulina e de determinados parâmetros bioquímicos de animais superalimentados durante a fase lactente. Além disso, o exercício Endurance se mostrou mais eficiente no controle das variáveis descritas acima, quando comparado ao exercício Hiit.
Statistical data of incidence and prevalence of Overweight/obesity have been growing over the years and this condition affects millions of people around the world, making it one of the great epidemics of the 21th century. Globalization and the increase in the availability of more palatable foods rich in saturated fat, added to the sedentary lifestyle, contribute directly to the development of this condition. Studies show that nutritional changes during critical stages of childhood development, such as lactation, are associated with the development of obesity, metabolic disorders, cardiovascular diseases, respiratory problems and diabetes. In addition, overweight/obesity can influence the reproductive function of the population causing fetal malformations, infertility, as well as maternal and fetal death. Considering physical exercise as a primary factor against overweight / obesity, we intend to evaluate whether physical exercise, in different intensities, is capable of influencing the reproductive health of overweight/obese individuals. Forty Wistar rats from normal litters (NL = 10) and small litters (SL = 4) were used and overweight/obesity was induced by litter reduction. The animals were trained for 8 weeks in protocols (Endurance and Hiit), mated at 90 days of age and euthanized on the 20th day of gestation. Metabolic, biochemical and reproductive datas were collected and analyzed. The animals of the overfeed groups trained Endurance and Hiit showed reduction with significant differences in body weight, consumption, insulin tolerance test and in some biochemical e reproductive parameters analyzed when compared to the control groups. However, there were no significant differences regarding nasoanal distance, relative organ weight, retroperitoneal and perigonadal fats, as well as oral glucose tolerance test, gestation index, implantation and fetal resorption in the studied groups. It was concluded that the physical training, in its different intensities, was not able to improve significantly the reproductive capacity of these animals, however, it was efficient to promote weight reduction, consumption, insulin sensitivity and certain biochemical parameters of supercharged animals during the phase. In addition, the Endurance exercise was more efficient in controlling the variables, described above, when compared to the Hiit exercise.

This thesis explored the interactions between psychological well-being and male fertility using the biopsychosocial model. The biopsychosocial model proposes that biological, psychological, and social processes interact and impact on health. These interrelationships were investigated in a sample of men undergoing fertility treatment and in a set of experiments using an animal model of stress. It is commonly thought that men with male factor infertility suffer more compared to men in couples with other infertility diagnoses, mainly due to the social stigma attached to being a man unable to father. The inter-relationships among diagnosis, psychological stress, and social environment were examined in men during a twelve month period of fertility treatment. It was found that men, regardless of diagnosis, showed signs of suffering over time and perceived some deterioration in their social environment that was at least partly caused by their psychological well-being at the start of treatment. To better understand how stress and reproductive processes interact, an animal stressor paradigm was developed. Male rats were exposed to a psychosocial cage change stressor (PCCS) where housing alternated every day between being alone, or in a new combination of two or three rats per cage for either 12 or 24 days. The four experiments showed that exposure to PCCS induced a mild physical stress response and consistent effects on reproductive parameters. It was concluded that the psychological and social aspects of the PCCS each have an impact on reproduction. This thesis has provided evidence of biopsychosocial links in the reproductive context supporting a biopsychosocial model of male fertility.

Background: Chlamydia trachomatis in women is a risk factor for tubal factor infertility and extra uterine pregnancies, but the impact of a C. trachomatis infection on male fertility is unclear. It is also hypothesized that persistent infection with C. trachomatis, or other microorganisms, might initiate/promote ovarian tumor development. The aims of the thesis were to study whether C. trachomatis serum antibodies in women and men had an impact on infertility diagnoses, semen characteristics, pregnancy rates and pregnancy outcomes; furthermore, to explore associations of C. trachomatis, and Mycoplasma genitalium, plasma antibodies with epithelial ovarian cancer and borderline ovarian tumors, as well as the presence of C. trachomatis bacteria, and other microorganisms, in ovarian tissues. Materials and methods: Papers I and II: 244/226 infertile couples were tested for serum C. trachomatis IgG, IgA, IgM and chlamydial Heat Shock Protein 60 (cHSP60) IgG antibodies. C. trachomatis IgG positive couples were also tested for C. trachomatis DNA in a urine sample. The follow-up period was 14-54 months. 244 spontaneously pregnant women were also tested for serum C. trachomatis IgG antibodies. Papers III and IV: Plasma samples from 291 women with epithelial ovarian cancer, borderline ovarian tumors and benign conditions, and plasma samples from 271 healthy controls, were analyzed for C. trachomatis IgG, IgA and cHSP60-1 IgG and M. genitalium IgG antibodies. Ovarian tissues from 186 women with benign ovaries, borderline ovarian tumors and epithelial ovarian cancer, as well as tissues from the contra lateral ovary in 126 women, were analyzed for the presence of C. trachomatis, M. genitalium, Neisseria gonorrhoeae, HPV and the polyoma viruses BKV and JCV with nucleic acid amplification tests. Results: Papers I and II: The prevalence of C. trachomatis IgG antibodies was higher among infertile than fertile women, and there were 9 couples with ongoing C. trachomatis infections. In men, C. trachomatis IgG and IgA antibodies were associated with a reduced likelihood to achieve pregnancy for the couple, as well as lower sperm concentration, reduced sperm motility and vitality, increased teratozoospermia index and the occurrence of leukocytes. C. trachomatis IgG and cHSP60 IgG antibodies in infertile women were associated with tubal factor infertility, but not with reduced pregnancy rates or outcomes. Paper III: cHSP60-1 IgG antibodies were associated with ovarian cancer belonging to the postulated type II pathogenetic pathway when plasma samples obtained more than one year prior to diagnosis were analyzed. M. genitalium IgG antibodies were associated with borderline ovarian tumors; however a statistical type 1 error cannot be excluded. Paper IV: None of the microorganisms studied were found in the ovarian tissue samples. Conclusions: C. trachomatis IgG and IgA antibodies in the man substantially decreases the chances of the infertile couple to achieve pregnancy, and are associated with subtle negative changes in semen characteristics. C. trachomatis IgG and cHSP60 IgG antibodies in the woman are risk factors for tubal factor infertility. Prospective plasma cHSP60-1 IgG antibodies are associated with type II ovarian carcinomas, but C. trachomatis bacteria, or the other microorganisms studied, could not be detected in benign, borderline or malignant ovarian tissues.

CoordenaÃÃo de AperfeiÃoamento de Pessoal de NÃvel Superior
Endometriose à uma doenÃa caracterizada pela presenÃa de glÃndulas e estroma endometriais for a da cavidade uterina e do miomÃtrio. Clinicamente pode causar tumores, dor (dor pÃlvica crÃnica, dispareunia e dismenorrÃia) e infertilidade. EvidÃncias correntes indicam que as cÃlulas endometriais ectÃpicas durante a menstruaÃÃo (menstruaÃÃo retrÃgrada) jogadas a cavidade peritoneal em mulheres com endometriose, implantam e proliferam ectÃpicamente no peritÃnio e em outros ÃrgÃos, uma aÃÃo que està associada com a mobilizaÃÃo de cÃlulas do sistema imune para a cavidade peritoneal e com uma profunda resposta imune e local. Um aumento na quantidade do lÃquido peritoneal à um achado caracterÃstico na endometriose e està associado com a presenÃa aumentada de cÃlulas imunes como os macrÃfagos assim como um sem numero de substancias solÃveis derivadas daqueles macrÃfagos como prostaglandinas, interleucinas, TNF, fatores de crescimento e espÃcies reativas de oxigÃnio. à provÃvel por isso que novas medicaÃÃes para essa doenÃa evoluam no futuro prÃximo, para isso devem-se elucidar os mecanismos de todas essas substÃncias na patogÃnese da endometriose. Os objetivos desse trabalho sÃo verificar: (i) os efeitos de drogas inibidoras seletivas da enzima ciclooxigenase tipo 1 (COX-1) responsÃvel pelos eventos fisiolÃgicos do organismo e pela induzida COX-2, envolvida nos processos inflamatÃrios; (ii) os efeitos do L-NAME, um antagonista competitivo do Ãxido nÃtrico assim como a atividade da NO-sintase avaliada pelo ensaio da citrulina marcada (3H-labelled citrulline from labelled L-arginine) e (iii) os efeitos da drogas moduladoras de TNF-alfa (Talidomida e pentoxifilina) sobre o desenvolvimento da endometriose experimental e suas repercussÃes sobre a dor e a fertilidade em ratas. Endometriose experimental foi desenvolvida em ratas Wistar e os animais foram divididos em grupos testes. O tratamento foi dado do 5o ao 15o dia da induÃÃo da endometriose para verificar os efeitos sobre o crescimento dos endometriomas avaliados pelo peso Ãmido e histopatologia e agudamente 30 minutos antes do estÃmulo nociceptivo para avaliar a dor (contorÃÃes abdominais) e a fertilidade investigada pela percentagem de ratas grÃvidas e pelo numero de embriÃes por corno uterino. Aspirina (30mg/kg - um inibidor de seletividade intermediÃria entre as COXs); piroxicam (1mg/kg) e indometacina (2mg/kg), um inibidor seletivo especÃfico da COX-1 e nabumetona (5 e 15mg/kg) e meloxicam (0,4mg/kg) inibidor seletivo relativo da COX-2 foram usados por via oral. Todos os tratamentos realizados diminuÃram significativamente a dor quando avaliadas pelo teste de contorÃÃes. A mÃdia dos pesos Ãmidos dos endometriomas (g%) sÃo mostrados (Controle endometriose: 0,595Â0,085; Aspirina: 0,122Â0,019; piroxicam: 0,766Â0,35; indometacina: 2,05Â0,96 e para Nabumetona 5mg: 0,52 0,032; Nabumetona 15mg: 0,135Â0,03 e meloxicam: 0,387Â0,04). Com relaÃÃo à fertilidade, a percentagem de ratas grÃvidas foi: Controle endometriose, 40%; controle intacto, 100%; Falso-operado, 100%; Indometacina, Zero%; meloxicam, 60%; Aspirina, 60% e Nabumetona 5 e 15, 50 e 58% respectivamente. Os tratamentos com Aspirina e Nabumetona diminuÃram significativamente o desenvolvimento dos endometriomas assim como contribuÃram para o alivio da dor e incrementaram a fertilidade. Estes resultados sugerem o papel da COX-1 e -2 na fisiopatologia da dor relacionada a endometriose assim como a infertilidade e o crescimento dos endometriomas. A atividade da sintase de Ãxido nÃtrico realizada atravÃs da citrulina marcada dada em pmol de citrullina/mgxproteÃna/min à expressa nos endometriomas. A iNOS no 5o dia: 1,94+0,5; 10o dia: 2,46Â0,2 e no dia 15: 1,17Â0,3 assim como com a cNOS que diminui de forma tempo-dependente (5 dia: 2,48Â0,7; 10 dia: 1,8Â0,19; e dia 15: 0,78Â0,3). Essa diminuiÃÃo da atividade da cNOS à provavelmente devida a descamaÃÃo endometrial que ocorre normalmente com a evoluÃÃo da doenÃa assim como devida a fibrose que circunda os endometriomas e o aumento da iNOS pelo processo inflamatÃrio peritoneal encontrado na endometriose. O uso do L-NAME tambÃm fez diminuir os pesos Ãmidos dos endometriomas assim como melhorou a fertilidade e aliviou a dor de forma dose-dependente. A Pentoxifilina (30mg/Kg/day) administrada entre o 5 e o 14 dia da induÃÃo da endometriose foi efetiva na diminuiÃÃo da expressÃo da sintase de Ãxido nÃtrico, ambas constitutiva como induzida. Os resultados desse estudo sugerem o envolvimento do Ãxido nÃtrico no desenvolvimento da endometriose experimental assim como nas suas repercussÃes: dor e infertilidade. Os nÃveis peritoneais de TNF-alfa em ratas intactas foram de 28,95Â1,18ng/ml. Os nÃveis de TNF-alfa aumentaram no lÃquido peritoneal de ratas endometriÃticas de forma tempo dependente. Drogas que modulam o TNF foram efetivas em reduzir o crescimento de endometriomas experimentais: Controle: 0,595Â0,085g%; pentoxifilina (30 mg/Kg): 0,06Â0,008g%; talidomida (5mg/Kg): 0,20Â0,049g% e dexametasona (0,2mg/Kg): 0,145Â0,02g%. Essas drogas tambÃm aliviaram a dor e incrementaram a fertilidade. Esses resultados sugerem o envolvimento do TNF na fisiopatologia da endometriose.
Endometriosis is a disease characterized by the presence of endometrial glands and stroma out of the uterine cavity and of the myometrium. It may cause tumor, pain (chronic pelvic pain, dyspareunia and dysmenorrhea) and infertility. Currently available evidence indicates that endometrial cells misplaced during menses into the peritoneal cavity in women with endometriosis, implant and proliferate in the ectopic locations, an action that is associated with mobilization of the immune cells into the peritoneal cavity and a profound local and systemic immune response. An increase in the amount of peritoneal fluid is a characteristic finding in endometriosis and associated with improved presence of immune cells like macrophages as well as a wide range of soluble substances derived from those macrophages like prostaglandins, interleukins, TNF, growth factors and reactive oxygen species. It is likely the role of medication for this disease will expand in the future. Also the mechanisms of all these substances must be elucidated in the pathogenesis of endometriosis. The purpose of this study is to verify: (i) the drugs effects that selectively inhibits one of the enzymes ciclooxigenase type 1 (COX-1) responsible for the physiologic events of the organism and the induced COX-2, involved in the inflammatory process; (ii) the effects of L-NAME, a competitive antagonist of nitric oxide as well as NO synthase actvity assayed by 3H-labelled citrulline from labelled L-arginine and (iii) the effects of TNF-alfa modulating drugs (Thalidomide and pentoxifilline) on the development of experimental endometriosis and on its related pain and infertility in female rats. Experimental endometriosis was developed in female Wistar rats and the animals were divided into tests groups. The treatment was given from the 5th to the 14th day of endometriosis induction to verify the effects on growth of endometriomas evaluated by its wet weight and histopathology and acutely 30 minutes before the nociceptive stimulus in order to evaluating pain (writhing test) and fertility was assessed through the percentage of pregnant rats. Aspirin (30mg/kg - an inhibitor of intermediate selectivity among COXs); piroxicam (1mg/kg) and indomethacin (2mg/kg), specific selective inhibitors of COX-1 and nabumetone (5 and 15mg/kg) and meloxicam (0.4mg/kg) relative selective inhibitors against COX-2 were used per os. All the accomplished treatments decreased significantly the pain as evaluated by the writhing test. The mean wet weights of the endometriomas (g%) were as shown [Endo control: 0.595 Â 0.085; Aspirin: 0.122 Â 0.019; piroxicam: 0.766 Â 0.35; indomethacin: 2.058 Â 0.96 and for Nabumetone 5mg: 0.252 Â 0.032; Nabumetone 15mg: 0.135 Â 0.03 and meloxicam: 0.387 Â 0.04]. As to fertility, the percentage of pregnant animals were as follows: Endo control, 40%; intact control, 100%; Sham operated, 100%; Indomethacin, Zero%; meloxicam, 60%; Aspirin, 60% and Nabumetone 5 and 15, 50 e 58% respectively. The treatments with Aspirin and Nabumetone had decreased the development of the endometriomas significantly as well as contributed to the relief of the pain and increasing fertility. These results suggest the role of COX-1 and -2 in the pathophysiology of endometriosis related pain, fertility and on its growth. NO synthase actvity assayed by 3H-labelled citrulline from labelled L-arginine. The nitric oxide synthase was expressed as pmol of citrulline/mg protein/min. The endometriomas expressed iNOS at the: 5th day: 1.94 + 0.5; 10th day: 2.46 Â 0.2 and day 15: 1.17 Â 0.3 as well as cNOS that decreased in a time-dependent way (5th day: 2.48 Â 0.7; 10th day: 1.8 Â 0.19; and day 15: 0.78 Â 0.3). This decreasing activity of cNOS was probably found by the endometrial shedding that occurs normally in the course of this disease as well as by the fibrosis that surrounds the endometriomas and the increasing iNOS by the inflammatory peritoneal and tissue reaction that is frequently found in endometriosis. The use of L-NAME also decreased the wet weight of endometriomas as well as ameliorates the pain and fertility in a dose dependent fashion. Pentoxifylline (30mg/Kg/day) administered subcutaneously for 10 consecutive days during the established phase of endometriosis (days 5-14 post induction) was effective in decreasing the expression of nitric oxide synthase, both induced and constitutive. The results of the present study as those previously shown suggest the involvement of nitric oxide in the development of experimental endometriosis. TNF-alfa modulating drugs proved to reduce the mean weights of endometrial implants, obtained at day 15 of endometriosis induction with pentoxifylline (30 mg/Kg), thalidomide (5mg/Kg) and dexamethasone (0.2mg/Kg) treated rats (control: 0.595 Â 0.085g%; pentoxifylline: 0.06 Â 0.008g%; thalidomide: 0.206 Â 0.049g% and dexamethasone: 0.145 Â 0.02g%). The results of the present study suggest the involvement of TNF-alfa in the pathophysiology of experimental endometriosis. The peritoneal levels of TNF-alfa evaluated in intact rats showed 28.95 Â 1.18ng of TNF-alfa/ml. The levels of TNF-alfa increased in the peritoneal fluid in a time dependent way after the peritoneal implant (endometriotic rat). These drugs also attenuated pain and increased fertility.

Between 10-15% of women suffer from polycystic ovary syndrome (PCOS), making it the most common cause of female infertility. Clinical features of PCOS include high circulating levels of ovarian androgens (T and A4), anovulation and obesity. The aetiology of this reproductive endocrinopathy is likely to be multifactorial, through the interplay of genetics, epigenetics and environmental factors. Primate research into sexual behaviour development noted that fetally androgenised monkeys developed symptoms like those of PCOS. There are now multiple animal models of PCOS using primates, sheep, rats and transgenic mice. The investigations described in this thesis use rodent models to examine the role of androgens in the pathogenesis of female infertility. An attempt to generate a granulosa cell specific androgen receptor knockout mouse model will first be described, followed by several studies into the developmental programming of female Wistar rat infertility and metabolism by steroid hormones. Initial investigations showed that testosterone proprionate (TP) administered to female rats during different windows of fetal and neonatal life alters the reproductive and metabolic axes of the adult animals. Fetal plus neonatal TP exposure led to complete ovarian dysgenesis, while postnatal exposure produced a PCOS-like phenotype. Animals which received TP postnatally were heavier and had an increased proportion of primordial follicles in their ovaries by postnatal day (pnd) 90 of life. Evaluation of this PCOS model showed that neonatally androgenised rats had ovarian follicles with larger antra and a greater ovarian stromal compartment. In addition, these animals were heavier when compared to controls. However, unlike human studies, neonatally androgenised rats showed no differences in circulating gonadotrophin or ovarian androgen levels. Nor did they show any programming effect of neonatal TP upon the theca interna by pnd 90. Further investigations to narrow the windows and dose of TP required to produce a PCOS phenotype showed that TP administered in an early window of neonatal life, between postnatal days (pnd) 1-6 not only led to anovulation, but potentially reprogrammed the hypothalamic-pituitary axis, as there was minimal gonadotrophin response to reduced ovarian negative feedback (inhibin B and estradiol) in these rats. Neonatal TP also affected the rat metabolic axis with adult animals becoming heavier after weaning without any change in food intake. Animals developed mesenteric and retroperitoneal obesity along with insulin resistance (IR). Increased hepatic glucocorticoid turnover and altered adipokine expression were also noted in neonatally androgenised females, possibly contributing to the pathogenesis of obesity. No effect of TP dose upon the severity of infertility or metabolic abnormalities in adult animals was observed. To delineate which features of the rat PCOS model resulted from androgenic, estrogenic or corticosteroid action, a final study used administration of different steroids during the early window of postnatal life: TP, estradiol valerate (EV), dihydrotestosterone (DHT), dehydroepiandrosterone (DHEA) and dexamethasone (DEX). The anovulatory PCO-like phenotype observed with TP was also seen in animals which received EV, but not those which received DHT, DHEA or DEX. TP and EV treatment also resulted in a reduction of ovarian follicle numbers and activated follicle proportions, with an increase in primordial follicle proportions. Although glucose tolerant, animals treated with TP and EV were highly IR. Unlike dexamethasone, DHT and DHEA also produced IR in adult animals, to a lesser extent than TP and EV. Taken collectively, the results described in this thesis demonstrate that the PCOS-like phenotype observed in the neonatally androgenised female rat is likely to be due to the estrogenic actions of testosterone, potentially through as yet unknown epigenetic mechanisms. The programming of the metabolic components described may additionally be due to the actions of androgens. Furthermore, these studies demonstrate a novel estrogenic effect of neonatal steroids upon primordial follicle populations and show that the neonatally androgenised rat may be a rational PCOS model in a poly-ovulatory species.

Orientadores: Sidney Glina, Paulo Cesar Rodrigues Palma
Dissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Ciencias Medicas
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Resumo: Introdução: As operações para correção das hérnias inguinais podem ocasionar lesões nos ductos deferentes levando à infertilidade, sendo que a incidência destas lesões varia de 6,65% a 26,7%. As telas de polipropileno têm sido utilizadas em indivíduos jovens nas últimas décadas. Este material induz processo inflamatório e fibrose adjacente conferindo maior reforço à parede posterior do canal inguinal, entretanto não se conhece claramente a repercussão desta reação sobre a histologia e fisiologia do ducto deferente. Objetivo: Este estudo avaliou o comprometimento ou não do ducto deferente de ratos expostos à tela de polipropileno. Método: Quarenta ratos, wistar, machos com três meses de idade, foram submetidos à dissecação bilateral dos ductos deferentes seguidos do implante da tela de polipropileno em um dos lados, tomando-se o lado contra-lateral como seu controle. Os animais foram sacrificados após 90 (Grupo I) e 120 (Grupo II) dias de pós-operatório, e os ductos deferentes, epidídimos e testículos avaliados histopatologicamente por microscopia óptica. Na análise estatística foram utilizados os testes de ?t? de Student e comparação de proporções. Resultados: Houve reação inflamatória tipo corpo estranho em todos os segmentos do ducto deferente exposto à tela e a montante desta, mas em nenhum no lado controle. Não houve obstrução completa do lúmen dos ductos deferentes expostos à tela de polipropileno em nenhum dos animais. O lúmen do ducto deferente apresentou dilatação a montante da tela em relação ao lado controle tanto no grupo I, que apresentou área média de 0,46441 mm², quanto no grupo II, área média de 0,37052 mm², no segmento controle do grupo I a área média foi de 0,2394 mm² e grupo II de 0,20789 mm², e as diferenças entre estes segmentos foram significativas (grupo I: p=0,002 / grupo II: p=0,044). Nos segmentos à montante da tela dos grupos I e II evidenciou-se a presença de espermatozóides no lúmen em 100% dos cortes histológicos, e nos controles de ambos os grupos a presença de espermatozóides ocorreu em 73,68%, sendo estas diferenças significativas (grupo I: p=0,009 / grupo II: p=0,022). A espessura da parede do ducto deferente foi significativamente menor nos segmentos expostos à tela (média de 0,15952 mm / p=4,348E-5) e a montante destes (média de 0,08348 mm / p=9,719E-10) em relação ao ducto deferente controle (média de 0,29968 mm) no grupo I. No grupo II, a espessura média foi de 0,294 mm e 0,1553 mm respectivamente nos segmentos exposto à tela e a montante desta, e 0,3034 mm no controle, apresentando significância apenas entre o segmento a montante da tela e o controle neste grupo (p=0,001). Houve perda do aspecto festonado da mucosa do ducto deferente em todos os segmentos à montante e nenhum no lado controle em ambos os grupos. Nenhum animal apresentou alterações significativas dos epidídimos e testículos, exceto um que apresentou atrofia testicular por provável torção do cordão espermático durante o ato operatório. Conclusão: A tela de polipropileno induz reação inflamatória, a qual resulta em alterações histológicas do ducto deferente que levam à dilatação e represamento de espermatozóides à montante da tela, assim como diminuição da espessura da parede do ducto deferente nos segmentos expostos à tela e à montante dela
Abstract: Background: Vas deferens lesions, leading to infertility, may occur in 6.65% to 26.7% of inguinal herniorraphy. Tension-free herniorraphy using polypropylene mesh, which induces an inflammatory process and adjacent fibrosis, strengthening the posterior inguinal wall, has become very popular. However its histological and physiological effects are not completely known. Objective: This study intended to evaluate the effects of polypropylene mesh in the rat vas deferens. Method: Forty, three-month-old male Wistar rats underwent bilateral vas deferens dissection, followed by mesh implantation on one side. The contra-lateral side was the control. The animals were sacrificed after 90 (Group I) and 120 (Group II) postoperative days. The vas deferens, epididymis and testicles underwent histopathological assessment. Statistics methods used were the ?t? test and the compare proportion. Results: A foreign body reaction occurred after implantation in all animals. Control presented no reaction. The mean lumen dilation in regions before mesh was 0.46441mm² and 0.37052mm², in Group I and II, respectively, and all sections presented spermatozoids. The mean dilation in control rats of Groups I and II was 0.2394mm² and 0.20789mm², respectively and spermatozoids were present in 73,68%. Those differences, between experimental and control animals were significant (group I: p=0,002 / group II: p=0,044). In Group I the average wall thickness of vas deferens segments exposed to mesh was 0.15952mm and the average wall thickness segments before mesh was 0.08348mm while the average wall thickness of control vas deferens was 0.29968mm (p=9,719E-10). In Group II, the mean wall thickness of vas deferens exposed to mesh and before mesh segments was 0.294mm and 0.1553mm, respectively, while the control was 0.3034mm (p=0,001). Mucosal fold loss occurred in all segments before mesh but none in the control. The epididymis and testicles presented no alterations. Conclusion: Polypropylene mesh induces a foreign body reaction with histological alterations in the vas deferens that cause functional obstruction with dilation and spermatozoid repression
Mestrado
Cirurgia
Mestre em Cirurgia

The chromatin integrity in sperm cells is vital for successful pregnancy. In this

study DNA-damage was evaluated in sperm cells from 50 men attending In Vitro Fertilization

(IVF) or Intra Cytoplasmic Sperm Injection (ICSI) treatment. Male semen samples were

purified with a two-shift gradient before the sperm cells were treated with the Halosperm® Test

Kit and evaluated for DNA-damage. The samples were divided in two groups according to DNAFragmentation

Index (DFI) of 30 % and the results correlated with fertilization, cleavage and

pregnancy rate. Men with DFI ≥ 30 % had a higher fertilization and pregnancy rate and a lower

cleavage rate compared to men with DFI ≤ 30 %. The conclusions were that fertilization in vitro

may be independent of the degree of DNA-damage, the embryonic development could be

seriously disrupted by damaged sperm cells, and the pregnancy rate showed no correlation to a

DFI threshold of 30 %.

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The prevalence of obesity has increased in recent decades in many countries, and harmful effects to the body can begin in childhood and persist into adulthood. Obesity is associated with disturbances of reproductive function in women and in female rodents, such as early onset of puberty, change in menstrual / estrous cycle and infertility, with impaired ovulation or anovulation (lack of ovulation). Angiotensin II (Ang II) appears to exert effects on reproduction and obesity, contributing to the development of the deleterious effects of obesity and affecting pre-ovulatory surge of luteinizing hormone (LH), estradiol and progesterone, thus reducing ovulation in adult rats. The objective of this study was to evaluate losartan effect, an antagonist of Ang II AT1 receptor, administrated in adulthood, in follicular development of adult Wistar rats with obesity induced by cafeteria diet. After weaning at 21 days of life, female Wistar rats were divided into 02 groups: control (CTL) that received standard chow diet; Cafeteria (CAF) received the cafeteria diet. From 70 days of life began losartan administration by gavage. The CTL group received water in gavage (CTL) and CAF group was separated into 02 groups, CAF (which received water in gavage) and CAF + LOS (who received losartan in gavage), in total 03 groups were performed. 05 animals were used per group. Euthanasia was performed on the first proestrous after 30 days of administration of losartan or water. The retroperitoneal, perigonadal and subcutaneous fat were removed and weighed. Morphological analysis of ovaries was performed, proceeding to count the number of primary follicles, secondary, antral and mature follicles per ovary. Was also collected blood sample for determination of FSH, LH, prolactin and progesterone. Body weight and the weight of the 03 fats were measured, and the number of antral follicles were higher in group CAF in relation to the group CTL (p <0.001). However, FSH and LH levels were lower in the CAF animals compared to the animals of the group CTL (p <0.001). The administration of losartan normalized the body weight and accumulation of retroperitoneal and subcutaneous fat as well as the number of antral follicles. Thus, we suggest that the use of the antagonist of Ang II AT1 receptor, losartan, in adulthood, can improve follicular development in females with cafeteria diet-induced obesity and, can be, in the future, an coadjuvant drug in the treatment of infertility associated to obesity.
A prevalência da obesidade aumentou nas últimas décadas em vários países, tendo efeitos prejudiciais ao organismo que podem iniciar na infância e persistir até a vida adulta. A obesidade está associada a distúrbios da função reprodutiva, em mulheres e em fêmeas de roedores, como início precoce da puberdade, alteração do ciclo menstrual/estral e infertilidade, com alterações da ovulação (disovulias) ou anovulação (ausência da ovulação). A Angiotensina II (Ang II) parece exercer efeitos sobre a reprodução e a obesidade, contribuindo para o desenvolvimento dos efeitos deletérios da obesidade e afetando o pico pré-ovulatório do hormônio luteinizante (LH), progesterona e estradiol, reduzindo, consequentemente, a ovulação em ratas adultas. O objetivo deste estudo foi avaliar o efeito do losartan, um antagonista do receptor AT1 da Ang II, sobre o desenvolvimento folicular de ratas Wistar adultas, com obesidade induzida pela dieta de cafeteria. Após o desmame, aos 21 dias de vida, ratas Wistar foram separadas em 02 grupos: controle (CTL) que recebeu ração padrão e Cafeteria (CAF) que recebeu a dieta de cafeteria. A partir dos 70 dias de vida iniciou-se a administração de losartan por gavagem. O grupo CTL recebeu água na gavagem (CTL) e o grupo CAF foi separado em 02 grupos, CAF (que recebeu água na gavagem) e CAF+LOS (que recebeu losartan na gavagem), totalizando 03 grupos. Foram utilizados 05 animais por grupo. A eutanásia foi realizada no primeiro proestro após 30 dias da administração de losartan ou de água. As gorduras retroperitoneal, perigonadal e subcutânea foram retiradas e pesadas. Foi realizada a análise morfológica dos ovários, procedendo-se a contagem do número dos folículos primários, secundários, antrais e maduros por ovário. Também foi coletado o sangue total para dosagens de FSH, LH, Prolactina e Progesterona. O peso corporal, bem como, o peso das 03 gorduras foram avaliados, e, o número de folículos antrais foi estatisticamente maior no grupo CAF em relação ao CTL (p<0,001). Todavia, as concentrações de FSH e LH foram menores nos animais CAF em relação ao CTL (p<0,001). A administração do losartan normalizou o peso corporal e o acúmulo das gorduras retroperitoneal e subcutânea, bem como, o número de folículos antrais. Dessa forma, sugerimos que o uso do antagonista do receptor AT1 da Ang II, o losartan, na vida adulta, possa contribuir para melhorar o desenvolvimento folicular em fêmeas com obesidade induzida pela dieta de cafeteria e, futuramente, ser uma droga coadjuvante no tratamento da infertilidade associada à obesidade.

Uvod: Infertilitet pogađa 10-15% parova reproduktivnog doba. Vanetesna oplodnja (VTO) je najefikasniji vid tret-mana infertiliteta, ali uprkos značajnom napretku stopa uspeha VTO u proseku iznosi oko 30% po ciklusu. Glavnim razlogom neuspeha smatra se neadekvatan kvalitet embriona, dok se pretpostavlja da u 10-20% slučajeva razlog neuspeha leži u neadekvatnoj receptivnosti uterusa. Na osnovu inicijalnih istraživanja histeroskopija, koja predstvalja zlatni standard u dijagnostici i tretmanu patologije kavuma uterusa, se često izvodi u svakodnevnoj kliničkoj praksi kako bi se povećala uspešnost VTO. Uprkos širokoj primeni i dalje ne postoji dovoljno kvalitetnih dokaza o realnoj ulozi histeroskopije na ishod VTO kako kod patoloških stanja kavuma tako i rutinski, pre prvog ili rekurentnog pokušaja VTO. Cilj disertacije bio je da se utvrdi uticaj sprovođenja histeroskopije na ishod VTO, ustanovi učestalost prethodno neprepoznate patologije kavuma uterusa, kao i da se ispitaju stavovi pacijenata o primeni rutinske histeroskopije pred VTO. Materijal i metode: Istraživanje je sprovedeno u Kliničkom centru Vojvodine, u formi prospektivne studije u dve sukcesivne etape od 01.01.2015. do 01.04.2017. U prvoj etapi poređen je ishod VTO kod pacijentkinja kojima pred postupak VTO nije sprovedena histeroskopija (Grupa A), pacijentkinja kod kojih je dobijen uredan nalaz histeroskopije pred postupak VTO (Grupa B) i pacijentkinja gde je pred postupak VTO dobijen patološki nalaz kavuma na histeroskopiji koji je u istom aktu tertian (Grupa C). Druga etapa istraživanja predstavljala je randomiziranu kontrolisanu studiju (RCT – randomised controlled trial). Nakon verifikacije urednog ultrazvučnog nalaza pred prvi postupak VTO, pacijentkinje su randomizirane u Grupu A2 kojima pred postupak VTO nije sprovedena histeroskopija i Grupu B2 kojima je pred postupak VTO sprovedena rutinska histeroskopija. Statistička analiza sprovedena je upotrebom odgovarajućeg softvera (JMP Ver. 9). Poređeni su podaci o osnovnim karakteristikama pacijenata, toka i ishoda ciklusa VTO. Primarni parametar ishoda bila je stopa kliničke trudnoće po embriotransferu. Pored analize ishoda primarno konstruisanih grupa, urađena je analiza i naknadno konstruisanih subgrupa, kao i predikcioni model uspeha VTO baziran na logističkoj regresiji. Rezultati: Studija je uključila 253 pacijentkinje (52 pacijentkinja iz Grupe A, 50 iz Grupe B, 50 iz Grupe C, 51 iz Grupe A2 i 50 iz Grupe B2). Nije postojala statistički značajna razlika u karakteristikama pacijentkinja, parametrima ovarijalne rezerve, broju dobijenih jajnih ćelija ni drugim parametrima toka postupka VTO među posmatranim grupama. U prvoj etapi istraživanja dobijena je statistički značajno (p=0,013) veća stopa kliničkih trudnoća kod pacijentkinja kojima je pred postupak VTO sprovedena histeroskopija - 50 % za Grupu B i 42% za grupu C u odnosu na 30,77% kod pacijentkinja bez histeroskopije (Grupa A), bez statistički značajne razlike među histeroskopskim grupama. U drugoj etapi istraživanja stopa kliničkih trudnoća prilikom upotrebe rutinske histeroskopije pred prvu VTO (Grupa B2) iznosila je 46% naspram 31,37% kod pacijentkinja bez histeroskopije pred prvu VTO (Grupa A2), iako uočena razlika nije dostigla statističku značajnost (p =0,089), uz relativan rizik (RR) za ostvarivanje kliničke trudnoće nakon primene histeoskopije uiznosio od 1,47 (95% CI 0,88-2,43) (p=0,13). Analizon subgrupa kod 100 pacijentkinja sa rutinski sprovedenom histeroskopijom pred VTO i 103 pacijentkinje bez histeroskopije pred VTO, dobijena je statistički značajnao veća stopa kliničkih trudnoća (48% naspram 31,07%, istim redom), uz RR od 1,54 (95% CI 1,08-2,20) (p=0,013), kao i stopa tekućih trudnoća od RR 1,49 (CI 1,01-2,19) (p= 0,039). Analiza ukupnog uticaja izvođenja histeroskopije pred VTO dobila je statistički značanjno veću stopu kliničkih trudnoća po ET za grupu histeroskopije uz RR 1,48 (CI 1,06-2,07) (p=0,017). Histeroskopijom je nakon urednog ultrazvučnog nalaza ustanovljeno postojanje patološkog nalaza kod 34,65% pacijenata i to 22,7% major patologije i 11,88% minor patologije kavuma. Nije postojala statistički značajna razlika u uspehu VTO u odnosu na sam nalaz histeroskopije. 98,67% pacijenata podržalo je rutinsku upotrebu histeroskopije pred prvi postupak VTO, dok je 83% pacijenata podržavlo rutinsku upotrebu histeroskopije pred svaki postupak VTO. U finalnom predikcionom modelu se uz AUC od 0,748 jedino postojanje visokokvalitetnog embriona uz odnos šansi (OR) 7,91 (95% CI 1,80-56,06; p=0,0047), transfer blastociste uz OR 3,80 (95% CI 1,90-7,98; p=0,0001) i izvođenje histeroskopije pred VTO uz OR 2,13 (95% CI 1,14-4,08, p=0,0169) pokazalo statistički značajnim prediktorima trudnoće. Diskusija: Studija je dobila pozitivan uticaj histeroskopije na ishod postupka VTO, iskazan pre svega povećanjem stope kliničkih trudnoća nakon sprovođenja histeroskopije (bilo da je na histeroskopiji nađen uredan ili patološki nalaz). Dodatna prednost histeroskopije predstavljala je i i detekcija prethodno nepropoznate patologije kavuma. Umeren efekat na ukupno poboljšanje stope kliničkih trudnoća prilikom rutinskog sprovođenja histeroskopije pred prvu VTO, koji je statističku značajnost dostigao tek analizom subgrupa u skladu je sa nalazima novijih dobro dizajniranih studija koji donekle limitiraju nekritičku upotrebu histeroskopije. Biološko objašnjenje potencijalnog pozitivnog uticaja histeroskopije najverovatnije leži u detekciji i tretmanu prethodno nepropoznate patologije kavuma, olakšavanju procedure embriotransfera, kao i humoralnim i molekularnim promenama koje nastaju u endometrijumu kao posledica odgovarajuće histeroskopske traume a koji su u dosašanjim istraživanjima apostrofirani kao faktori koji mogu povećati receptivnost uterusa. Zaključak: Histeroskopija je efikasna, bezbedna i visoko prihvatljiva procedura koja dovodi do povećanja uspeha VTO u standardnim kliničkim indikacijama (prethodnog neuspelog postupka VTO i sumnje na patološki nalaz kavuma uterusa) bilo da se na samoj histeroskopiji nađe uredan ili patološki nalaz. Rutinska primena histeroskopije pred prvi postupak VTO se na osnovu rezultata studije ne može smatrati apsolutno opravdanom usled statistički nedovoljno značajnog povećanja stope kliničke trudnoće. Uzevši u obzir visoku prihvatljivost od strane pacijenata i najverovatniji pozitivan efekat na stopu trudnoće primena rutinske histeroskopije pred prvu VTO bila bi opravdana ukoliko se implementira koncept ambulantne histeroskopije.
Introduction: Infertility affects 10-15% of all couples. In vitro fertilisation (IVF) is the most effective method of infertility treatment, but despite a significant improvement, success rate of IVF is still around 30% per cycle. The main reason for the IVF failure is inadequate embryo quality, but in 10-20% of cases the cause of IVF failure lies in impaired uterine receptivity. Based on earlier studies hysteroscopy, gold standard in the diagnosis and treatment of uterine cavity pathology, is often performed to increase IVF success. Despite its wide use, there is lack of high quality evidence regarding real contribution of hysteroscopy on IVF outcome in situations of uterine cavity pathology or routinely prior to first IVF or after recurrent implantation failure. The aim of this dissertation was to determine the influence of performing hysteroscopy on IVF outcome, as well as the incidence of previously unrecognized uterine pathology, and to examine patient's attitudes about performing routine hysteroscopy prior to IVF. Material and methods: The research was conducted in a prospective manner in two successive stages at Clinical Center of Vojvodina from 01.01.2015. until 01.04.2017. During first stage of the study IVF outcome was compared between patients who did not have a hysteroscopy prior to IVF (group A), patients with normal hysteroscopic finding prior to the IVF (Group B) and patients with abnormal hysteroscopic findings prior to IVF which was treated at the same time (Group C). The second stage of the study was a randomized controlled trial (RCT). After verification of normal ultrasound findings prior to the first IVF, patients were randomized to group A2 in who me hysteroscopy was not performed and group B2 who had routine hysteroscopy prior to first IVF. Statistical analysis was carried out using the appropriate statistical software (JMP Ver. 9). Patient characteristics, course and outcome of IVF cycle were compared between groups. The primary outcome was clinical pregnancy rate (CPR) per embryotransfer. In addition to analyzing the IVF outcomes in primarily defined groups, subgroup analysis was also performed, as well as IVF success pre-diction model based on logistic regression. Results: The study included 253 patients (52 patients in Group A, 50 in Group B, 50 in Group C, 51 in Group A2 and 50 in Group B2). There was no statistically significant difference in patient characteristics, ovarian reserve parameters, number of retrieved oocytes, or other relevant parameters of IVF course between the observed groups. In the first stage of the study there was statistically significant (p = 0.013) higher clinical pregnancy rate in patients who had a hysteroscopy before IVF - 50% for Group B and 42% for group C versus 30,77 % in patients without hysteroscopy before IVF (Group A), without statistically significant difference between hysteroscopic groups. In the second stage of the study, routine hysteroscopy prior to first IVF (Group B2) led to clinical pregnancy rate 46% versus 31.37% in patients without hysteroscopy prior to first IVF (Group A2), although without statistical significance (p = 0.089. Relative risk (RR) for achieving clinical pregnancy after performing hysteroscopy was 1.47 (95% CI 0.88-2.43) (p = 0.13). Subgroup analysis of 100 patients with routinely performed hysteroscopy before IVF and 103 patients without hysteroscopy prior to the IVF showed statistically significant higher rates of clinical pregnancies (48% versus 31.07%, in the same order), with RR of 1.54 (95% CI 1.08-2.20), (p = 0.013), and for ongoing pregnancies RR was 1.49 (95% CI 1.01-2.19) (p = 0.039). Overall effect of performing hysteroscopy prior to IVF resulted in a statistically significant increase in the clinical pregnancy with RR 1.48 (95% CI 1.06-2.07) (p = 0.017). After normal ultrasound finding hysteroscopy revealed 34.65% of pathological finding, 22.7% of major and 11.88% of minor pathology of the cavity). There was no statistically significant difference in IVF outcome based on hysteroscopy findings. 98.67% of patients supported the routine use of hysteroscopy before the first IVF procedure, while 83% of patients supported the routine use of the hysteroscopy before every IVF procedure. In the final prediction model, with the AUC of 0.748, only the presence of high quality embryos with odds ratio (OR) 7,91 (95% CI 1,80-56,06; p=0,0047), blastocyst transfer with OR 3,80 (95% CI 1,90-7,98; p=0,0001) and performing hysteroscopy prior to IVF with OR 2,13 (95% CI 1,14-4,08, p=0,0169) proved to be statistically significant predictors of pregnancy. Discussion: The study shoved a positive influence of hysteroscopy on the IVF outcome by increasing clinical pregnancy rate after performing hysteroscopy (whether hysteroscopy revealed normal or pathological finding). Additional benefit of hysteroscopy was detection of previously unrecognized uterine pathology. A moderate effect on the overall improvement in clinical pregnancy rate with use of routine hysteroscopy, which reached statistical significance only by subgroup analysis, is in line with findings of recent well designed studies that somewhat limit the noncritical use of hysteroscopy. A biological explanation of the potential positive effect of hysteroscopy is most likely due to detection and treatment of the previously unrecognized uterine pathology, facilitating embryotransfer procedure, as well as the humoral and molecular changes that occur in the endometrium as a consequence of the hysteroscopic trauma. Those changes were hypothesized as factors that can increase uterine receptivity by numerous research. Conclusion: Hysteroscopy is an effective, safe and highly acceptable procedure that increases IVF success when performed for accepted clinical indications (previous IVF failures, pathological findings of uterine cavity), whether hysteroscopy reveals normal or pathological finding. The routine use of hysteroscopy prior to first IVF based on this study can not be considered justified since increase in clinical pregnancy rate did not reach statistical significance. Given the high acceptance of this concept by the patients and moderate but probable positive effect on IVF outcome, implementation of routine hysteroscopy prior to first VTO would be justified only in office hysteroscopy setting.

Uppsatsen undersöker hur ”SOU 2016:11 Olika vägar till föräldraskap” formulerar problemet med surrogatmoderskap, vilka subjektspositioner som kan urskiljas i utredningen och analyserar den ut ett kritiskt postkolonialt feministiskt perspektiv.   Utredningen utgår ifrån mänsklig värdighet, autonomi och barnets bästa vilket uppsatsen belyser får konsekvenser för hur problemet med surrogatarrangemang porträtteras olika beroende på om surrogatmodern och barnet är från det globala syd eller det globala nord. Utgångspunkterna är centrala när surrogatmodern och barnet från det globala nord lyfts men får inte samma roll gällande surrogatmodern och det barn hon föder. Vidare undersöks hur subjektspositioner formuleras utifrån emotionell/icke-emotionell, sårbar/icke-sårbar, skyddsvärd/icke-skyddsvärd och blottlägger hur den svenska surrogatmodern och det inrikesfödda barnet konstrueras som emotionella, sårbara och skyddsvärda vilket skiljer sig från hur den utländska surrogatmodern och det utrikesfödda barnet konstrueras. Dessa omges att tystnad vilket får konsekvenser för hur de porträtteras och för vilka åtgärder som föreslås.   Uppsatsen bidrar till en utveckling av tidigare forskning med ett tillägg av nyckelordet skyddsvärd för att undersöka konstrueringen av surrogatmödrar. Uppsatsen visar att begreppet är relevant genom att materialet visat på delvis konstuering av emotionell och sårbar men inte skyddsvärd och att detta påverkat resultat och åtgärder. Vidare stärker uppsatsen tidigare forskning som menar att utgångspunkten mänsklig värdighet verkar för en avhumanisering av surrogatmödrar från det globala syd samt att synen på biologi varierar beroende på var ett barn föds. Uppsatsen bygger vidare på tidigare forskning som menar att uttryckt rationalitet och brist på emotion resulterar i en avhumanisering, genom att blottlägga hur det kan räcka med att subjektspositionerna möts av tysthet för att uppnå samma resultat. Detta arbete belyser hur olika subjektspositioner framställs olika i den statliga utredningen och hur koloniala idéer hänger kvar och påverkar underlag till svensk lagstiftning, samt blottlägger vilka konkreta konsekvenser detta kan få för lagförslag och för surrogatmödrar i det globala syd samt de barn de föder.

碩士
中山醫學大學
醫學研究所
104
Objective：The objective of this study is to elucidate the effect of systemic and local antioxidant status upon sperm function of unexplained infertility (UI) couples in in vitro fertilization (IVF) cycles with density gradient centrifugation (DGC) preparation method. The relevance of the antioxidant status in the body and semen to the fertilization and embryo development in IVF cycles was explored. Methods and Materials：A total of 60 semen samples from UI couples undergoing IVF treatment were collected for the observational study. The skin carotenoid status and seminal total antioxidant capacity (TAC) were measured as biomarkers of systemic and local antioxidant status, respectively. The density gradient centrifugation method was used for sperm preparation in IVF cycles. Results：The results indicated that the DGC method for sperm preparation in IVF cycles increased the levels of reactive oxygen species but was not associated with increasing oxidative injury of spermatozoa. Both skin carotenoid status and seminal TAC were associated with fertilization rates and good embryo rates in IVF cycles. Conclusion and Suggestion：The systemic carotenoid status and seminal plasma TAC of male partners of UI couples are closely related to fertilization and embryo development in IVF cycles. This result implies that antioxidant supplement to the body and seminal plasma might benefit the sperm function in this patient group. Our data might provide more evidence about the role of oxidative stress in male partners of UI couples and a possible new strategy for the management of UI couples.

Des facteurs masculins sont identifiés dans près de la moitié des cas d’infertilité. À ce jour, les tests évaluant la fertilité masculine demeurent peu prédictifs de la survenue d’une grossesse. Dans le but de pallier cette lacune, nous avons mis au point deux nouveaux tests mesurant l’intégrité de l’ADN et le temps de survie des spermatozoïdes. Nous avons effectué une étude prospective portant sur 42 couples infertiles suivis en fécondation in vitro (FIV). Le spermogramme a été effectué selon les critères de l’Organisation Mondiale de la Santé (OMS) et le temps de survie des spermatozoïdes exposés à un détergent cationique a été mesuré en observant la mobilité sous microscope. L’intégrité de l’ADN des spermatozoïdes a été vérifiée par la nouvelle méthode de marquage radioenzymatique et par analyse de la structure de la chromatine (SCSA). Tous les tests ont été réalisés sur la partie des échantillons de sperme non utilisée par la clinique de fertilité. Le projet a été approuvé par le comité d’éthique du Centre Hospitalier Universitaire de Montréal (CHUM) et les patients ont préalablement signé un formulaire de consentement éclairé. L’analyse des paramètres du spermogramme et de l’intégrité de l’ADN n’a montré aucune différence statistiquement significative entre les données chez les couples avec ou sans grossesse. Cependant, le taux de grossesse biochimique était statistiquement plus élevé chez les couples dont le temps de survie des spermatozoïdes était long (>250 s) comparativement à ceux dont ce temps était court (≤250 s): 66% vs 27% respectivement (p<0,05). Les taux de grossesse clinique et d’implantation étaient aussi plus élevés, mais les différences n’atteignaient pas le seuil de signification statistique. Nos résultats confirment que le spermogramme et la mesure de la fragmentation de l’ADN des spermatozoïdes ne sont pas de bons facteurs prédictifs des résultats de la FIV. Par contre, le test de survie des spermatozoïdes serait un meilleur indicateur de la possibilité d’une grossesse en FIV. L’amélioration de sa spécificité et un plus grand nombre de sujets sont nécessaires avant de proposer son application en clinique de fertilité.
Male factors are known to be involved in almost half of the couples consulting for infertility. To date, the tests for evaluating male fertility are poor predictors of pregnancy. We developed two new tests to evaluate sperm function: a sperm survival test and a new method to measure sperm DNA integrity. This prospective study was conducted on 42 infertile couples undergoing in vitro fertilization (IVF). Assessment of sperm parameters was done according to the World Health Organization (WHO) criteria, and sperm survival upon exposure to a cationic detergent was measured by observing motility under the microscope. Sperm DNA integrity was verified by our new radioenzymatic method as well as by the sperm chromatin structure analysis (SCSA) method. All testing was performed on a remainder aliquot of the semen samples. The study was approved by the ethics committee of the Centre Hospitalier Universitaire de Montréal (CHUM), and informed consent was obtained before inclusion. Neither conventional semen analysis, nor sperm DNA fragmentation showed statistically significant difference between conception and non-conception cycles. However, the biochemical pregnancy rate was statistically higher in couples where the sperm survival time was long (>250 s) compared to short (≤250 s): 66% vs. 27% respectively, (p < 0.05). The clinical pregnancy rate and implantation rate were also higher but the differences did not reach statistical significance. Our study confirms that conventional semen analysis and the assay for sperm DNA integrity are not reliable indicators of IVF outcome. In contrast, our new sperm survival test seems to be a better predictor of the pregnancy rate after IVF. Improvement of its specificity and a larger cohort of patients are necessary before proposing its regular application in IVF clinics.

The diploma thesis deals with the change of the family in today's society, its roles, and the ways of cohabitation which replace the family. It evaluates the significance of marriage and parenthood for children and for their future lives. It describes the difficulties related to responsible parenthood and the role of parents during their children's adolescence. It evaluates the mentioned issues from the point of view of Christian ethics. It also touches on the problems of the current society in terms of infertility and assisted reproduction as well as ethical issued connected with them.