Introdução: A resistência insulínica (RI) decorrente da obesidade está relacionada a distúrbios hormonais que afetam o sistema reprodutor. Leptina e grelina são hormônios que regulam o balanço energético; porém, informações acerca da relação destes hormônios com a infertilidade são escassas. A dieta de baixo índice glicêmico (BIG) parece exercer impacto positivo sobre as alterações metabólicas decorrentes da RI. Objetivo: Verificar o efeito de uma dieta hipocalórica de baixo índice/carga glicêmica sobre parâmetros antropométricos e metabólicos, níveis de grelina e leptina e desfechos reprodutivos em mulheres inférteis com excesso de peso candidatas à fertilização in vitro (FIV). Métodos: Ensaio clínico randomizado. Foram analisadas vinte e seis mulheres inférteis com obesidade Grau I ou II ou pré-obesidade associada à circunferência da cintura aumentada. As pacientes foram alocadas no grupo Dieta Hipocalórica de BIG, ou no grupo Controle (manutenção do hábito alimentar) e acompanhadas por 12 semanas. Parâmetros avaliados: peso corporal, índice de massa corporal (IMC), percentual de gordura (%G), glicose, insulina, HOMA-IR, lipídios séricos, hormônios reprodutivos, grelina acilada, leptina, dose de gonadotrofinas, número e qualidade oocitária e embrionária, taxa de fertilização e de gestação. Resultados: Houve redução de 5,5% do peso corporal e também do IMC (p < 0,001), do %G (p = 0,002), dos níveis de glicose (p = 0,034) e de leptina (p = 0,013) no grupo BIG quando comparado ao grupo controle. Houve um aumento de 18% nos níveis de grelina no grupo BIG quando comparado ao controle, mas esse aumento não foi significativo (p > 0,05). O grupo BIG obteve 85,4% mais oócitos coletados, quando comparado ao grupo controle (7,75 ± 1,44 vs. 4,18 ± 0,87, respectivamente, p = 0.039) no ciclo de FIV. Não houve diferença entre os grupos na dose de gonadotrofinas, na qualidade oocitária e embrionária, e na taxa de fertilização. Três (21,4%) pacientes do grupo BIG apresentaram gestação espontânea durante o acompanhamento, gerando três nascidos vivos. Conclusões: A perda de 5,5% do peso corporal através da dieta hipocalórica BIG foi capaz de melhorar parâmetros antropométricos, metabólicos, reprodutivos e os desfechos de FIV, quando comparado às mulheres que mantiveram o peso corporal. Estes resultados dão sustentação à recomendação clínica de aconselhar mulheres com sobrepeso ou obesas a perderem peso através de uma dieta balanceada, preferencialmente com baixo índice/carga glicêmica, antes de serem submetidas a procedimentos de reprodução assistida. / Introduction: Insulin resistance (IR) resulting from obesity is related to hormonal disorders that affect reproductive system. Leptin and ghrelin are hormones that regulate energy balance; however, the relationship of these hormones with infertility is not clear. The low glycemic index (LGI) diet seems to exert a positive impact on obesity and metabolic changes resulting from IR. Objective: To verify the effect of a hypocaloric diet with low glycemic index/load on anthropometric and metabolic parameters, ghrelin and leptin levels and reproductive outcomes in overweight and obese infertile women candidates to in vitro fertilization (IVF). Methods: Randomized clinical trial. Twenty six infertile women with grade I and II obesity, or pre-obesity with increased waist circumference were analysed. Patients were assigned to hypocaloric LGI diet group or control group (maintenance of usual diet), and followed the protocol for 12 weeks. Parameters evaluated: body weight, body mass índex (BMI), body fat percentage (%BF), glucose, insulin, HOMA-IR, serum lipids, reproductive hormones, leptin, acylated ghrelin, gonadotrophin doses, number and quality of oocytes and embryos, fertilization and pregnancy rates Results: There was a 5.5% weight loss and also a reduction in BMI (p < 0.001), BF% (p = 0.002), glucose (p = 0.034) and leptin levels (p = 0.013) in the LGI group compared to control. There was a 18% increase in ghrelin levels in the LGI group compared to control, but this increase was not significant (p > 0.05). The LGI diet group had 85.4% more oocytes retrieved compared to control group (7.75 ± 1.44 vs. 4.18 ± 0.87, respectively, p = 0.039) in the IVF cycle. The gonadotrophin dose, oocyte and embryo quality, and fertilization rate were similar between groups (p > 0.05). Three (21.4%) patients in the LGI group experienced spontaneous pregnancy during the follow-up, generating three live births. Conclusion: The 5.5% weight loss trough the hypocaloric LGI diet was able to improve antropometric, metabolic, reproductive and IVF outcomes when compared with women that not lose weight. These results support the clinical recommendation to advise overweight and obese women to lose weight through a balanced diet, preferably with low glycemic index/load, prior to be submitted to assisted reproduction technologies.

Submitted by Mario BC (mario@bc.ufrpe.br) on 2017-05-04T12:12:28Z No. of bitstreams: 1 Pabola Santos Nascimento.pdf: 865543 bytes, checksum: 47ce2118a4d55e4ada483756d00b9ae6 (MD5) / Made available in DSpace on 2017-05-04T12:12:28Z (GMT). No. of bitstreams: 1 Pabola Santos Nascimento.pdf: 865543 bytes, checksum: 47ce2118a4d55e4ada483756d00b9ae6 (MD5) Previous issue date: 2014-08-28 / The use of sexed semen for in vitro embryo production (IVP) is a potentially effective means for obtaining the progeny with predetermined sex. For years, animal owners wanted a methodology that pre determine the sex of offspring for their herds. Rates of cleavage, morula and blastocyst seem to be affected not only by the damages caused by sexing, but it is believed that the bull factor, and other aspects that have not been fully elucidated, directly influencing its successful use. The aim of this study was to evaluate the influence of type of semen (sexed /conventional) and a bull factor over blastocyst rates when submitting bovine oocytes obtained from slaughterhouse to IVP, and also compare these results with analyzes of sperm kinetics. Oocytes (n = 959) were matured, fertilized with sexed and non-sexed semen from three 5/8 Girolando bulls. A straw of each type of semen was assessed with use of "computer-assisted semen analysis" (CASA) and fluorescence microscopy. Three replicates were performed during the experiment. Data were analyzed by SPSS 16.0 statistical program employing analysis of variance (ANOVA). The Student t test was used to detect differences between groups. The Chi-square test was used to analyze the results of embryo production. For all analyzes, values were considered significant (P <0.05). The results differed between sexed (21.10%) and non-sexed semen (31.06%) to blastocysts production. We conclude that in the present work sexed semen was less efficient in producing blastocyst when comparing non-sexed semen of the same bulls and when comparing semen types (sexed and non-sexed) from the same bull. Analyses of sperm kinetics and fluorescent probes were compatible with the fertilizing potential of samples of sexed and non-sexed semen of 5/8 Girolando bulls. / O uso de sêmen sexado em conjunto com a produção in vitro (PIV) de embriões é um meio potencialmente eficiente na obtenção da descendência com sexo predeterminado. Há anos, os proprietários dos animais desejaram uma metodologia que pré determinasse o sexo da prole para seus rebanhos. As taxas de clivagem, mórula e blastocisto parecem ser afetadas não só pela debilitação provocada na sexagem, mas acredita-se que pelo reprodutor em questão, e por outros fatores que não foram ainda totalmente elucidados, influenciando diretamente na sua utilização com sucesso. O objetivo desse trabalho foi avaliar a influencia do tipo de sêmen (sexado/convencional) e do touro, nas taxas de blastocisto na PIV de oócitos bovinos obtidos em matadouro, e ainda comparar esses resultados com as análises da cinética espermática. Oócitos (N= 959) foram maturados, fertilizados com sêmen sexado e convencional de três touros da raça 5/8 Girolando. Uma palheta de cada tipo de sêmen foi avaliada com uso da “computer-assisted sêmen analysis” (CASA) e da microscopia de fluorescência. Foram realizadas três repetições durante o experimento. Os dados foram analisados pelo programa estatístico SPSS 16.0 empregando-se a análise de variância (ANOVA). O teste t-Student foi usado para detectar diferenças entre os grupos. O Qui-quadrado foi utilizado para análise dos resultados da produção de embriões. Para todas as análises, os valores foram considerados significativos (P< 0,05). Os resultados diferiram entre o sêmen convencional (31,06) e sexado (21,10%) para produção de blastocisto. Quando comparamos a produção de blastocisto individualmente nas amostras de sêmen sexado (27,69%; 17,93% e 25,56%, touros 1, 2 e 3 respectivamente) percebemos que T2 < T1 e T1=T3 e T2=T3. Concluimos que no presente trabalho o sêmen sexado foi menos eficiente na produção de blastocisto quando comparado ao sêmen convencional de forma geral e do mesmo touro. As análises da cinética espermática bem como as sondas fluorescentes foram compatíveis com o potencial fertilizante das amostras de sêmen sexado e convencional em touros da raça 5/8 Girolando.

Submitted by Marlene Santos (marlene.bc.ufg@gmail.com) on 2016-04-05T15:03:43Z No. of bitstreams: 2 Dissertação - Ariany Rafaella Neto Silva - 2015.pdf: 893987 bytes, checksum: 388b303a349c43a6a3fe0cc14e611756 (MD5) license_rdf: 19874 bytes, checksum: 38cb62ef53e6f513db2fb7e337df6485 (MD5) / Approved for entry into archive by Luciana Ferreira (lucgeral@gmail.com) on 2016-04-05T15:26:45Z (GMT) No. of bitstreams: 2 Dissertação - Ariany Rafaella Neto Silva - 2015.pdf: 893987 bytes, checksum: 388b303a349c43a6a3fe0cc14e611756 (MD5) license_rdf: 19874 bytes, checksum: 38cb62ef53e6f513db2fb7e337df6485 (MD5) / Made available in DSpace on 2016-04-05T15:26:45Z (GMT). No. of bitstreams: 2 Dissertação - Ariany Rafaella Neto Silva - 2015.pdf: 893987 bytes, checksum: 388b303a349c43a6a3fe0cc14e611756 (MD5) license_rdf: 19874 bytes, checksum: 38cb62ef53e6f513db2fb7e337df6485 (MD5) Previous issue date: 2015-08-04 / Fundação de Amparo à Pesquisa do Estado de Goiás - FAPEG / This study was developed with the objective of evaluating the response of bovine embryos produced in vitro (PIV) the addition of resveratrol (Resv) to recultivo to embryos vitrified (Experiment 1-E1) and medium Holding for fresh embryos (Experiment 2-E2) to check the maintenance or improvement in the quality of these embryos. In E1 PIV vitrified bovine embryos were heated and recultivados in the medium containing 0.5 μM of resveratrol, evaluating the re-expansion rates, hatching and cell quality by analysis of TUNEL. In E1, the hatching rate of recultivo more than 24 hours after (P < 0.05) in the Resv Group (37,7 vs. 19,1%). The rate of embryos according to the stage of development did not show difference between treatments. Even in E1, for fresh embryos PIV the total number of cells (NTC) was superior to that found in the vitrified embryos (131,6 vs 88,8 and 82,7%) , as well as the NCA (number of apoptotic cells) was lower (6,3 ± 0,8) compared with control groups and resveratrol (15,8 ± 1.2 and 13,5 ± 1,0), which did not differ among themselves in these variables. No difference was observed in embryos degenerates and reactive oxygen species (ROS) and Glutathione (GSH). In E2 bovine embryos were fresh bottled PIV in 0,25 ml straws with a holding in the presence (TR6 and TR10) or absence (TH6 and TH10) of resveratrol and kept on warming plate to 36° C in different times, six and 10 hours, followed by the unloading spouts and recultivo until 24 hours. Hatching rates at 24 hours of embryos treated with antioxidant tended to be greater than in the control. Bx TR10 rates was higher (51,2%) the TR6 (30,8%) (P < 0.05), the other stages of development were similar, the NTC was lower in TH10. The index of EROS was superior in TH10 (23,4126 ± 1,5661). The values of the GSH TR6 (95,2208) were larger than TH6 (30,7594) (P < 0.05), the TR10 issuing GSH (49,5330 ± 6,5332) did not differ from TH10 group (47,2044) (P > 0,05). / Esse estudo foi desenvolvido com os objetivos de avaliar a resposta de embriões bovinos produzidos in vitro (PIV) à adição de resveratrol (Resv) aos meios de recultivo para embriões vitrificados (Experimento 1 - E1) e Holding para embriões frescos (Experimento 2 - E2) para verificar a manutenção ou melhora na qualidade desses embriões. No E1 embriões bovinos PIV vitrificados foram aquecidos e recultivados em meio contendo 0,5 μM de resveratrol, avaliando-se as taxas de reexpansão, eclosão e qualidade celular através da análise de TUNEL. No E1, a taxa de eclosão após 24 horas de recultivo foi superior (P < 0.05) no grupo Resv (37,7 vs 19,1%). Na taxa de embriões de acordo com o estágio de desenvolvimento não apresentaram diferença entre tratamentos. Ainda no E1, para os embriões PIV frescos o número total de células (NTC) foi superior ao encontrado nos embriões vitrificados (131,6 vs 88,8 e 82,7%), assim como o NCA (número de células apoptóticas) foi menor (6,3 ± 0,8) em relação aos grupos controle e resveratrol (15,8 ± 1,2 e 13,5 ± 1,0), que não diferiram entre si nessas variáveis. Não foi observada diferença nas taxas de embriões degenerados e emissão de espécies reativas ao oxigênio (EROS) e glutationa (GSH). No E2 embriões bovinos PIV frescos foram envasados em palhetas de 0,25mL com meio holding na presença (TR6 e TR10) ou ausência (TH6 e TH10) de resveratrol e mantidos sobre placa aquecedora a 36°C em diferentes tempos, seis e 10 horas, seguido do desenvase e recultivo até 24 horas. As taxas de eclosão às 24hs dos embriões tratados com antioxidante tenderam a ser maior que no controle. As taxas de Bx TR10 foi superior (51,2%) ao TR6 (30,8%) (P < 0,05), os demais estágios de desenvolvimento apresentaram-se semelhantes. O NTC foi menor no TH10, assim como a MCI. O índice de EROS foi superior no TH10 (23,4126 ± 1,5661). Os valores de GSH do TR6 (95,2208) foram maiores que TH6 (30,7594) (P < 0,05), no TR10 a emissão de GSH (49,5330 ± 6,5332) não diferiu do grupo TH10 (47,2044) (P > 0,05).

O sucesso da fecundação e da sustentação do desenvolvimento embrionário subseqüente é dependente de atributos espermáticos relacionados à qualidade seminal. A produção in vitro de embriões em bovinos é uma ferramenta fundamental para a aceleração do ganho genético do rebanho, porém o uso comercial depende diretamente da melhoria do sistema de produções in vitrode embriões a um baixo custo. Técnicas que proporcionem melhoria da qualidade seminal possibilitando um incremento na produção in vitro de embriões e ainda a um baixo custo possuem apelo econômico. O laser de baixa potência emite ondas eletromagnéticas com efeitos biológicos. Este efeito é dependente dos parâmetros de irradiação e do tipo celular alvo. Trabalhos avaliando o laser de baixa potência em amostras seminais sugerem que seja utilizado para melhorar os atributos espermáticos e, conseqüentemente, a produção in vitro de embriões. Este estudo teve como objetivo avaliar o efeito do laser de baixa potência sobre a viabilidade espermática e averiguar o efeito desta estimulação na produção in vitro de embriões bovinos. Para isso, amostras comerciais de sêmen criopreservado foram descongeladas e submetidas à irradiação com laser He-NE. No experimento 1 foram testadas as potências de 0; 5; 7,5 e 10mW, por 5 ou 10 minutos de tratamento. As amostras de sêmen foram avaliadas imediatamente após a irradiação e após 30 minutos de incubação. Foram verificadas possíveis interações duplas (Potência*Duração do Tratamento, Potência*Tempo e Duração do Tratamento*Tempo) e tripla (Potência*Duração do Tratamento*Tempo). No experimento 2 foi avaliado o efeito da melhor combinação potência X duração dos tratamentos utilizados no Experimento 1, para irradiar o sêmen utilizado para a produção in vitro de embriões. O laser de baixa potência apresentou efeito sobre diversos atributos relacionados à viabilidade espermática. Este efeito foi dependente da potência e da duração do tratamento. Nas condições experimentais, a potência de 5mW com duração de 10 minutos de aplicação sugere efeitos positivos no aumento na viabilidade espermática. No entanto, o tratamento de espermatozóides nesta potência e duração do tratamento, não apresenta incremento nas taxas de produção nem na qualidade embrionária. / Success of fertilization and the maintenance of subsequent embryo development rely on sperm attributes related to seminal quality. The in vitro production of bovine embryos is an essential tool for genetic gain in the herd. However, its commercial use directly depends on improvement of the production system at a low cost. Inexpensive techniques which provide an increase of seminal quality, allowing the increment of in vitro production of embryos are of great interest to cattle breeding. Low-power laser emits electromagnetic waves with biological effects. These effects are dependent of irradiation parameters and target cell type. Studies with seminal samples irradiated by low potency laser suggest its use to improve several sperm features and, therefore, in vitro production of embryos. This study aimed to evaluate the effect of low potency laser on sperm viability and fertilization ability of irradiated sperm. Briefly, in Experiment 1, samples of frozen/thawed commercial semen were subjected to He-NE laser irradiation. Laser potencies of 0, 5, 7.5 or 10mW were tested for 5 or 10 min of treatment. Samples were evaluated immediately and 30 min after irradiation. Double (Potency*Treatment length, Potency*Time and Treatment length*Time) and triple interactions (Potency*Treatment length*Time) were assessed. Afterwards, in Experiment 2, the effect of the best combination of potency and exposure length on in vitro production of embryos was verified. Low potency laser affected several semen attributes related to sperm viability. This effect was dependent on laser potency and the length of treatment. In the experimental conditions tested in this study, 5mW potency combined to 10 min of exposure had appeared to have positive effect of increasing sperm viability. However, 5mW potency laser for 10 min sperm irradiation did not improve in vitro embryo production rates and bovine embryo quality.

Objetivo: Avaliar as evidências disponíveis comparando a eficácia da estimulação ovariana (EO) com uso de citrato de clomifeno (CC) e/ ou letrozol (LTZ) para reduzir o consumo de FSH, em relação à estimulação ovariana padrão (EOP). Métodos: Realizamos uma revisão sistematizada e meta-análise de ensaios clínicos randomizados (ECRs) que compararam os desfechos reprodutivos na fertilização in vitro. As buscas foram realizadas em onze bancos de dados eletrônicos e avaliamos manualmente a lista de referência dos estudos incluídos e revisões similares. Nós estratificamos os resultados separando os estudos baseados no agente oral utilizado (CC ou LTZ) e nas características da mulher incluída (em que se espera e em que não se espera má resposta ovariana). Os desfechos avaliados foram risco relativo (RR) para nascimento vivo, gravidez clínica, aborto, e taxa de cancelamento de ciclo, Peto Odds Ratio (OR) para síndrome de hiperestímulo ovariano (SHO), e diferença média (MD) para número de óocitos captados e consumo de FSH (ampolas). Resultados: Foram incluídos 22 estudos nesta revisão. Considerando o grupo de mulheres em que se espera má resposta, a evidência sugere que o uso de CC durante a estimulação ovariana resulta em similares taxas de nascidos vivos (RR= 0,9, IC95% = 0,6 a 1,2, evidência de moderada qualidade) e de gravidez clínica (RR= 1,0, IC95% = 0,8 a 1,4, evidência de moderada qualidade); o uso de LTZ não causa alteração significativa no número de oócitos captados (MD= -0,4, IC95% = -0,9 a +0,1, evidência de alta qualidade). Considerando os estudos que avaliaram mulheres em que não se esperava má resposta, a evidência sugere que o uso de CC reduz o número de oócitos captados (MD= -4,6, IC95%= -6,1 a -3,0, evidência de alta qualidade) e o risco de SHO (Peto OR= 0,2, IC95%= 0,1 a 0,3, evidência de moderada qualidade), enquanto os resultados são semelhantes para taxas de nascidos vivos (RR= 0,9, IC 95% = 0,7 a 1,1, evidência de moderada qualidade) e de gravidez clínica (RR= 1,0, IC95% = 0,9 a 1,2, evidência de alta qualidade). Para os demais desfechos a qualidade das evidências foi baixa ou muito baixa. Conclusões: A utilização de CC em mulheres em que se espera má resposta tem a vantagem de alcançar resultados reprodutivos semelhantes com redução dos custos. Para as demais mulheres, o uso do CC tem a vantagem adicional de reduzir o risco de SHO, mas também reduz o número de oócitos captados. Mais estudos seriam necessários para avaliar o efeito do LTZ com o mesmo propósito. Estudos futuros devem ter como objetivo estudar a taxa de gravidez cumulativa por oócito captado, insatisfação da paciente e aceitação para repetir o ciclo se não engravidar, que são dados importantes para a tomada de decisões clínicas. / Objective: To assess the available evidence comparing effectiveness of ovarian stimulation (OS) using clomiphene citrate (CC) and/or letrozole (LTZ) for reducing FSH consumption compared with standard OS. Methods: We performed a systematic review and meta-analysis of randomized controlled trials (RCTs) that compared the reproductive outcomes following in vitro fertilization. We searched eleven electronic databases and hand-searched the reference list of included studies and related reviews. We stratified the results separating the studies depending on the oral agent (CC or LTZ) and on the characteristics of the included women (expected poor ovarian response or other women). When combining the results of included studies, we assessed the relative risk (RR) for live birth, clinical pregnancy, miscarriage, and cycle cancelation, Peto Odds Ratio (OR) for OHSS, and mean difference (MD) for the number of oocytes retrieved and FSH consumption. Results: A total of 22 studies were included in this review. Considering women with expected poor ovarian response, the available evidence suggests that using CC for reducing FSH consumption during OS provide similar live birth (RR=0.9, 95%CI=0.6-1.2, moderate quality evidence) and clinical pregnancy rates (RR=1.0, 95%CI=0.8-1.4, moderate quality evidence); the use of LTZ doesn\'t cause a relevant change on the number of oocytes retrieved (MD=-0.4, 95%CI= -0.9 to +0.1, high quality evidence). Considering the studies evaluating other women, the available evidence suggests that using CC for reducing FSH consumption during OS reduces the number of oocytes retrieved (MD=-4.6, 95%CI=-6.1 to -3.0, high quality evidence) and the risk of OHSS (Peto OR=0.2, 95%CI=0.1-0.3, moderate quality evidence), while results in similar live birth (RR=0.9, 95%CI=0.7-1.1, moderate quality evidence) and clinical pregnancy rates (RR=1.0, 95%CI=0.9-1.2, high quality evidence). The quality of the evidence was low or very low for the other outcomes. Conclusion: The use of CC for reducing FSH consumption in women with expected poor ovarian response has the advantage of providing similar reproductive outcomes with reduced costs. For the other women, the use of CC for reducing FSH consumption has the additional advantage of reducing OHSS, but also reduces the total number of oocytes retrieved. More studies are necessary to evaluate the effect of LTZ for the same purpose. Future studies should aim on cumulative pregnancy per oocyte retrieval, patient dissatisfaction and agreement to repeat the cycle if not pregnant; which are important outcomes for clinical decisions.

Background: Since the initiation of assisted reproduction techniques, several studies has been performed to improve treatment results by development of culture conditions like embryo oil and culture media used. In this study, two embryonic oils from different companies, Nidoil and Ovoil were examined.Method: In this study, 47 human embryos were used. All embryos were donated for research purposes by couples who had been treated at the clinic in Uppsala University Hospital. The embryos were divided into two groups, one group was cultured with Ovoil and the other with Nidoil.Results: There was no difference between the two oils, the embryo quality was the same in both groups.CONCLUSION: The result was expected because both oils had the same composition and purity.

MSCAGR (Animal Science) / Department of Animal Science / See the attached abstract below

<p>Infertility is the disease of the reproductive system and is estimated to affect more than 10% of the people of reproductive age. Assisted reproductive technologies (ART) are methods designed to alleviate infertility problems. <i>In vitro </i>embryo production is part of most infertility treatments and the efficiency of ART is low due to the lack of reliable methods to measure embryo viability. In order to improve the success rate of ART procedures, the current study was designed to investigate the use of an optical analyzer technology known as the Biodynamic Imaging (BDI) system for viability assessment. BDI is a novel approach that is able to measure intracellular dynamic processes that are directly related to functional events. During a series of experiments, 13 different biomarkers of oocytes and embryos were monitored by the BDI microscope and used for machine learning and evaluation of BDI sensitivity. We monitored cellular mechanisms essential for proper embryo development such as (1) extrusion of first and second polar body (2) energy status and mitochondrial activity, and (3) viability of embryos with different cellular composition. We were able to identify several biomarkers that have the potential to indicate viability: (1) slope, (2) NSD, (3) Knee (4) Floor, and (5) R² could consistently differentiate between oocytes and embryos of different viability. In addition, the BDI microscope could successfully predict the energy status of embryos by identifying 4 biomarkers (Slope, Knee, Floor, and Dy). Finally, a lipidomic analysis was done to evaluate the lipid composition of oocytes with different cytoplasm integrities. This analysis demonstrated that there is a difference in lipid subclasses among oocytes with dark vs. light cytoplasm. The results indicate that the BDI is useful for viability assessment of oocytes and embryos and may be helpful for the improvement of the efficiency of assisted reproductive technologies.</p>

Micromotors – untethered, motile, microscopic devices – are implemented in this dissertation for two applications in the field of assisted reproductive technology. First, as synthetic motor units for individual sperm cells, representing a novel approach to counteract sperm immotility (asthenozoospermia), which is one of the most prevalent causes of male infertility. Second, as synthetic carriers of fertilized oocytes (zygotes) towards the realization of non-invasive intrafallopian transfer, representing a novel alternative to the current keyhole surgery (laparoscopy) approach to achieve early embryo transfer after in vitro fertilization. In both applications, magnetically actuated micromotors are utilized to capture, transport, and deliver individual cells in a reproducible, controllable manner. In comparison with established in vitro fertilization routines, the crucial advantage of employing micromotors for the manipulation of gametes, i.e. sperm and (fertilized) oocytes, lies in the potential transfer of decisive steps of the fertilization process back to its natural environment – the fallopian tube of the female patient – taking advantage of the untethered, non-invasive motion and manipulation capabilities of magnetic micromotors. When sperm motility can be restored with magnetic micromotors, sperm can travel to the oocyte under external actuation and control, and the oocyte does not need to be explanted for in vitro fertilization. However, if in vitro fertilization was necessary, fertilized oocytes can be transferred back to the fallopian tube by micromotors in a non-invasive manner, to undergo early embryo development in the natural environment. These novel concepts of micromotor-assisted reproduction are presented and investigated in this thesis, and their potential is analyzed on the basis of proof-of-concept experiments.:1 Introduction 6 1.1 Background and Motivation 6 1.2 Objectives and Structure of this Dissertation 9 2 Fundamentals 11 2.1 Micromotors Definition and Concept 11 2.2 Micromotors for Biomedical Applications 13 2.3 Magnetic Micropropellers 15 2.3.1 Theory 15 2.3.2 Implementation 20 2.4 Microfabrication: Direct Laser Writing 21 2.5 Assisted Reproductive Technology 23 2.5.1 In vitro Fertilization and Intracytoplasmic Sperm Injection 24 2.5.2 Embryo Transfer and Zygote Intrafallopian Transfer 25 2.5.3 The Sperm Cell and the Oocyte 26 2.6 Towards Micromotor-Assisted Reproduction 28 3 Materials and Methods 30 3.1 Fabrication of Microfluidic Channel Platforms 30 3.1.1 Tailored Parafilm Channels 30 3.1.2 Polymer Channels Cast from Micromolds 31 3.1.3 Tubular Channels to Mimic In vivo Ducts 32 3.2 Fabrication of Magnetic Micropropellers 32 3.2.1 Direct Laser Writing of Polymeric Resin 33 3.2.1.1 Design and Programming 33 3.2.1.2 Exposure and Development 35 3.2.1.3 In Situ Direct Laser Writing 35 3.2.2 Critical Point Drying 35 3.2.3 Magnetic Metal Coatings 36 3.2.4 Surface Functionalization 37 3.3 Sample Characterization 38 3.3.1 Optical Microscopy 38 3.3.2 Scanning Electron Microscopy 38 3.4 Cell Culture and Analysis 39 3.4.1 Sperm Cells 39 3.4.2 Oocytes 39 3.4.3 In vitro Fertilization 41 3.4.4 Hypoosmotic Swelling Test 44 3.4.5 Cell Viability Assays 44 3.5 Magnetic Actuation 45 3.5.1 Modified Helmholtz Coil Setup 46 3.5.2 MiniMag Setup 47 3.5.3 Experimental Procedure 48 3.5.3.1 Micromotor Performance Evaluation 48 3.5.3.2 Cell Transport Experiments 49 3.5.3.3 Cell Transfer Experiments 50 4 Micromotor-assisted Sperm Delivery 51 4.1 Micromotor Design and Fabrication 51 4.2 Actuation and Propulsion Performance 53 4.3 Capture, Transport, and Release of Sperm 56 4.4 Delivery to the Oocyte 59 4.5 Sperm Viability and the Ability to Fertilize 61 5 Micromotor-assisted Zygote Transfer 68 5.1 Micromotor Design and Fabrication 68 5.2 Actuation and Propulsion Performance 70 5.3 Capture, Transport, and Release of Zygotes 76 5.4 Transfer between Separate Environments 80 5.5 Zygote Viability and Further Development 82 6 Conclusions and Prospects 85 Appendix 87 Bibliography 93 List of Figures and Tables 108 List of Abbreviations and Terms 109 Theses 111 Selbstständigkeitserklärung 112 Acknowledgments 113 List of Publications 115 Curriculum Vitae 116

Inhibin A and B participate to regulation of gametogenesis. We investigated their applicability as a marker of gametogenesis of men fertility disorders. We monitored the levels of inhibins during the treatment. We interested in their paracrine activity, relationship in sera, follicular fluid and seminal plasma depending on cause of fertility failure. We studied the levels of inhibin B in serum and seminal plasma from 355 men treated for fertility failure, in the context of their andrological and immunological findings (quality of spermiogrammes and acrosome area). We monitored concentration of levels of inhibin A and B in serum and follicular fluids depending on cause of fertility failure, on course and treatment outcome. We took blood samples in the time of the oocytes collection, of the embryotransfer and early pregnancy. The follicular fluids were obtained during the oocytes collection. The levels of both of inhibins were measured by ELISA in all medium (serum, follicular fluid, seminal plasma). We confirm, that inhibin B is useful marker of spermatogenesis in men, but is necessary to examine patient in complex with determination of immunology profile or quality of acrosome. Seminal plasma is, in some indicated cases, more suitable diagnostics material. Similarly inhibin B in women seems to be...