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Efeito de diferentes momentos de inseminação artificial laparoscópica em programas de transferência de embriões correlacionados com o momento da ovulação em ovinosALMEIDA, Valdir Morais de 29 April 2013 (has links)
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Previous issue date: 2013-04-29 / The unpredictability of results in addition with a variable efficiency on embryo transfer (ET) in sheep can be related to the absence of timing definition on artificial insemination (AI) in ET programs by an asynchrony between ovulations and AI procedures in superovulted animals. The aim of this work, was to identify by laparoscopy the ovarian dynamics of ewes subjected to multiple ovulation embryo transfer program (MOET) and, based upon this knowledge, evaluate two ideal moments of laparoscopic AI, with cooled and frozen semen. In experiment 1, ten females, sheep were used, synchronized with intravaginal progesterone device (CIDR ®) (D0) and Superovulation induction by FSH-p (Folltropin ®), in eight decreasing administration doses every twelve hours, starting at D12. In D15 intravaginal devices have been removed and administered 200IU of equine chorionic gonadrotophin - eCG (Novormon ®, Intervet). The next day, 4μg of buserelin acetate – GnRH - was administered (Conceptal ®, Intervet). After estrous detection, observations of ovarian status laparoscopically were performed at 36, 40, 48, 52 and 56 hours after CIDR removal or until ovulations occurred. In experiment 2, twenty sheep females were used, synchronized and induced with identical protocol of experiment 1 and, after estrous detection, they were divided into four treatments defined as at the time of the Laparoscopic AI based upon removal of intravaginal device and on the type of semen used: T1-36 h/cooled semen; T2-48 h/cooled semen; T3-36 h/frozen semen and; T4-48 h/frozen semen. Five days after laparoscopic AI, collection of the embryos was performed by laparotomy and the recovered structures were evaluated and classified according to IETS (International Embryo Transfer Society) and transferred to recipients that had synchronized estrus by applying intravaginal device impregnated with 60 mg medroxiprogeterona-MAP (Progespon ®), for a period of 15 days, when the device was removed and 400UI of eCG (Folligon ®) was administered subcutaneously and embryos were transferred by semi-laparoscopy. Pregnancy diagnosis was performed by ultrasonography days after ET. There was a higher concentration (90%) of the time of ovulation at 48 hours after the device removal. At Fifty-two hours after device removal, all females from the experiment had ovulated. In the second experiment, no differences were seen in number of structures recovered among treatments with a mean of 11.15 ± 6.35. The number of viable embryos differed statistically between the T2 (8.4 ± 5.5) and T3 (4.0 ± 4.4), showing that cryopreserved sperm cell was not able to accomplishing fertilization when the laparoscopic AI was held at 36 h. It was concluded that females subjected to MOET show a fast follicular growth and ovulations occurred mostly 48 hours after Progesterone device removal. It is possible to suggest a single laparoscopic AI in commercial MOET programs in sheep. It was showed also a numerical superiority, when using cooled semen. / Os resultados pouco previsíveis e a eficiência variável da transferência de embriões (TE) em ovinos podem estar relacionados à ausência da definição do melhor momento da IA em programas de TE por assincronia entre as ovulações e os procedimentos da inseminação artificial (IA) em animais superovulados. Desta feita, buscou-se identificar o momento da ovulação, por meio da laparoscopia, de fêmeas ovinas submetidas a programa de múltipla ovulação e transferência de embriões-MOTE e, a partir destes conhecimentos, avaliar dois momentos da IA laparoscópica, com uso de sêmen resfriado e criopreservado. No experimento 1, dez fêmeas ovinas foram utilizadas, sincronizadas com uso de dispositivo vaginal de Progesterona (CIDR®) (D0) e indução da superovulação por administrações de FSH-p (Folltropin®), em oito doses decrescentes a cada doze horas, iniciando-se no D12. No D15 foram removidos os dispositivos intravaginais e administradas 200UI de gonadrotofina coriônica eqüina - eCG (Novormon®, Intervet). No dia seguinte, administrou-se 4μg de acetato de buserelina - GnRH (Conceptal®, Intervet). Após detecção do cio e jejum de 12 horas, foram realizadas as observações do status ovariano via laparoscópica, nos momentos 36, 40, 48, 52 e 56 horas pós-remoção do CIDR® ou até observação das ovulações. No experimento 2, vinte fêmeas ovinas foram utilizadas, sincronizadas e induzidas com protocolo idêntico ao do experimento 1 e, após detecção do cio e jejum de 12 horas, foram divididas em quatro tratamentos definidos quanto ao horário da IAL com base na remoção do dispositivo intravaginal e quanto ao tipo de sêmen utilizados: T1 – 36 h / sêmen resfriado; T2 – 48 h / sêmen resfriado; T3 – 36 h / sêmen criopreservado e; T4 – 48 h / sêmen criopreservado. Passados cinco dias da IAL, foi realizada a colheita dos embriões pela técnica de laparotomia e, as estruturas recuperadas foram avaliadas e classificadas segundo normas preconizadas pela IETS (International Embryo Transfer Society) e, transferidos para receptoras que tiveram estro sincronizado por meio da aplicação de dispositivo intravaginal impregnado com 60 mg de medroxiprogeterona – MAP (Progespon®), por um período de 15 dias, quando se deu a sua retirada e aplicação de 400UI de eCG (Folligon®) por via intramuscular e, receberam embriões pela técnica de semi-laparoscópia. O diagnóstico gestacional foi realizado por meio de ultrassonografia aos 35 dias após a TE. Observou-se maior concentração (90%) do momento das ovulações às 48 horas pós-retirada do dispositivo intravaginal. Cinquenta e duas horas pós-remoção do dispositivo intravaginal, todas as fêmeas do programa haviam ovulado. No segundo experimento, O número de estruturas viáveis, do T1 e T2, não variou significativamente, entretanto, aumento significativo (P 0,05), foi observado no percentual de confirmações de prenhezes entre os tratamentos, T3 = 9,1 ± 13,3 % e T4 = 49,2 ± 30,1 %, demonstrando que o uso da IAL com sêmen criopreservado, na condição de préovulação (36 h), não foi eficaz na fertilização das estruturas oócitarias. Observou-se que uma única IAL, foi suficiente para produção embrionária, com exceção do T3, alcançando-se melhores índices no T4, com fertilização de 62,5 ± 39,3 %, e confirmação ultrassonográfica de 49,2 ± 30,1 %, das estruturas produzidas. Conclui-se que fêmeas submetidas a programa de múltipla ovulação e transferência de embriões apresentam rápido crescimento folicular e maior concentração de ovulação às 48 horas pós-remoção dos dispositivos intravaginais, sendo possível indicar a utilização de uma única IAL com uso sêmen resfriado e, criopreservado em programas comerciais de MOTE em ovinos, evidenciando considerável ganho tecnológico e econômico.
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Visual SLAM and Surface Reconstruction for Abdominal Minimally Invasive SurgeryLin, Bingxiong 01 January 2015 (has links)
Depth information of tissue surfaces and laparoscope poses are crucial for accurate surgical guidance and navigation in Computer Assisted Surgeries (CAS). Intra-operative Three Dimensional (3D) reconstruction and laparoscope localization are therefore two fundamental tasks in CAS. This dissertation focuses on the abdominal Minimally Invasive Surgeries (MIS) and presents laparoscopic-video-based methods for these two tasks.
Different kinds of methods have been presented to recover 3D surface structures of surgical scenes in MIS. Those methods are mainly based on laser, structured light, time-of-flight cameras, and video cameras. Among them, laparoscopic-video-based surface reconstruction techniques have many significant advantages. Specifically, they are non-invasive, provide intra-operative information, and do not introduce extra-hardware to the current surgical platform. On the other side, laparoscopic-video-based 3D reconstruction and laparoscope localization are challenging tasks due to the specialties of the abdominal imaging environment. The well-known difficulties include: low texture, homogeneous areas, tissue deformations, and so on. The goal of this dissertation is to design novel 3D reconstruction and laparoscope localization methods and overcome those challenges from the abdominal imaging environment.
Two novel methods are proposed to achieve accurate 3D reconstruction for MIS. The first method is based on the detection of distinctive image features, which is difficult in MIS images due to the low-texture and homogeneous tissue surfaces. To overcome this problem, this dissertation first introduces new types of image features for MIS images based on blood vessels on tissue surfaces and designs novel methods to efficiently detect them. After vessel features have been detected, novel methods are presented to match them in stereo images and 3D vessels can be recovered for each frame. Those 3D vessels from different views are integrated together to obtain a global 3D vessel network and Poisson reconstruction is applied to achieve large-area dense surface reconstruction.
The second method is texture-independent and does not rely on the detection of image features. Instead, it proposes to mount a single-point light source on the abdominal wall. Shadows are cast on tissue surfaces when surgical instruments are waving in front of the light. Shadow boundaries are detected and matched in stereo images to recover the depth information. The recovered 3D shadow curves are interpolated to achieve dense reconstruction of tissue surfaces.
One novel stereoscope localization method is designed specifically for the abdominal environment. The method relies on RANdom SAmple Consensus (RANSAC) to differentiate rigid points and deforming points. Since no assumption is made on the tissue deformations, the proposed methods is able to handle general tissue deformations and achieve accurate laparoscope localization results in the abdominal MIS environment.
With the stereoscope localization results and the large-area dense surface reconstruction, a new scene visualization system, periphery augmented system, is designed to augment the peripheral areas of the original video so that surgeons can have a larger field of view. A user-evaluation system is designed to compare the periphery augmented system with the original MIS video. 30 subjects including 4 surgeons specialized in abdominal MIS participate the evaluation and a numerical measure is defined to represent their understanding of surgical scenes. T-test is performed on the numerical errors and the null hypothesis that the periphery augmented system and the original video have the same mean of errors is rejected. In other words, the results validate that the periphery augmented system improves users' understanding and awareness of surgical scenes.
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Caracterização do pico de LH após protocolo de curta e longa duração para IATF em ovelhas ciclando na contra estação reprodutiva / Characterization of the LH peak after short and long-term protocol for FTAI in cycling sheep out of the breeding seasonSoriano, Gabriela Azenha Milani 27 February 2015 (has links)
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Previous issue date: 2015-02-27 / The hypothesis tested in this study was that the time of permanence of progesterone implant (short vs. long) in FTAI protocol change the characteristics of the LH surge. Thus, the aim of this study was to determine the timing, duration and amplitude of the LH peak in sheep (Santa Inês x Texel) using short FTAI protocols (6 days) or long (12 days) in cyclic sheep out of the breeding season. We used 43 sheep (Texel-Te x Santa Inês-SI), sorted into two groups according to the time of progesterone release s vaginal device 1st use (Easy-Breed CIDR®, Pfizer, Brazil). In group 12 days (G-12, n = 19) in stage random of the estrous cycle received the insertion of CIDR (D0). On the day of implant removal (D12) was administered intramuscularly, 0.075 mg cloprostenol (Veteglan®, Hertape Calier, Brazil) and 300 IU equine chorionic gonadotropin (eCG, Novormon®, MSD Animal Health, Brazil). The group 6 (G-6, n= 24) received the same protocol of G-12, however the permanence of CIDR was 6 days. Approximately 50 hours after CIDR removal, both groups were inseminated at fixed time by artificial insemination by laparoscopy with frozen semen containing 200x106 sperm of a single player of Dorper. Approximately 40 days after the artificial insemination pregnancy diagnosis was performed using ultrasound device. Blood samples were taken every 4 hours from 10 animals per group, to measure LH and progesterone plasma concentrations by radioimmunoassay (RIA). From the 20 animals used for progesterone plasma concentration, the percentage of animals showing LH surge in G-6 was 70% (7/10) and the G-12 was 90% (9/10). The maximum amplitude of the LH peak did not differ between the G-6 and G-12 groups (P> 0.05) and the lengh of LH peak was 16,60±2,76 h in the G-6 and 16,40±2,40 h in G-12 (P>0,05). The interval between the implant removal and the LH surge was (P >0.05) in the G-6 27,42±2,76 h and 28,88±5,20 h in G-12. The overall pregnancy rate per group was not different (G-6 58,33% e G-12 52,63%). It was concluded that the permanence of progesterone (6 vs 12 days) did not alter the characteristics of the LH peak in sheep (St. Agnes x Texel) in the tropics, out of the breeding season. / A hipótese testada pelo presente trabalho foi que o tempo de permanência do implante de progesterona (curto vs longo), em protocolo de IATF, altera as características do pico de LH. Desta forma, o objetivo do presente estudo foi determinar o momento, duração e amplitude do pico de LH em ovelhas (Texel x Santa Inês) utilizando protocolos de IATF de curta (6 dias) ou longa duração (12 dias) em ovelhas ciclando na contra estação reprodutiva. Foram utilizadas 43 ovelhas (Texel-Te x Santa Inês-SI), divididas em dois grupos de acordo com o tempo de permanência do dispositivo vaginal de liberação de progesterona de 1° uso (Easy-Breed CIDR®, Pfizer, Brasil). No grupo 12 dias (G-12, n=19), em estádio aleatório do ciclo estral receberam a inserção do CIDR (D0). No dia da retirada do implante (D12) foram administrados, por via intramuscular, 0,075 mg de cloprostenol (Veteglan®, Hertape Calier, Brasil) e 300 UI de gonadotrofina coriônica equina (eCG, Novormon®, MSD Saúde Animal, Brasil). O grupo 6 (G-6, n=24) recebeu o mesmo protocolo do G-12, porém a permanência do CIDR foi de 6 dias. Aproximadamente 50 horas após a retirada do CIDR as ovelhas, de ambos os grupos, foram inseminadas em tempo fixo por laparoscopia com sêmen congelado contendo 200x106 espermatozóides, de um único reprodutor, da raça Dorper. Aproximadamente 40 dias após as inseminações artificiais foi realizado o diagnóstico de prenhez utilizando aparelho de ultra-som. Foram realizadas colheitas de sangue, de 4 em 4 horas em 10 animais por grupo, para mensuração das concentrações plasmáticas de LH e progesterona por radioimunoensaio (RIA). Dos 20 animais utilizados para mensuração das concentrações plasmáticas, a porcentagem de animais que apresentaram pico de LH no G-6 foi de 70% (7/10) e no G-12 foi 90% (9/10). A amplitude máxima do pico de LH não diferiu entre os grupos G-6 e G-12 (P>0,05) e a duração do pico de LH foi 16,60±2,76 h no grupo G-6 e 16,40±2,40 h no grupo G-12 (P>0,05). O intervalo entre a retirada do implante e o pico de LH foi (P>0,05) ocorreu 27,42±2,76 h no grupo G-6 e 28,88±5,20 h no G-12. A percentagem de prenhez total por grupo não diferiu (G-6 58,33% e G-12 52,63%). Conclui-se que o tempo de permanência da progesterona (6 vs 12 dias) não alterou as características do pico de LH em ovelhas (Texel x Santa Inês) nos trópicos, na contra estação reprodutiva.
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Caracterização do pico de LH após protocolo de curta e longa duração para IATF em ovelhas ciclando na contra estação reprodutiva / Characterization of the LH peak after short and long-term protocol for FTAI in cycling sheep out of the breeding seasonSoriano, Gabriela Azenha Milani 27 February 2015 (has links)
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Previous issue date: 2015-02-27 / The hypothesis tested in this study was that the time of permanence of progesterone implant (short vs. long) in FTAI protocol change the characteristics of the LH surge. Thus, the aim of this study was to determine the timing, duration and amplitude of the LH peak in sheep (Santa Inês x Texel) using short FTAI protocols (6 days) or long (12 days) in cyclic sheep out of the breeding season. We used 43 sheep (Texel-Te x Santa Inês-SI), sorted into two groups according to the time of progesterone release s vaginal device 1st use (Easy-Breed CIDR®, Pfizer, Brazil). In group 12 days (G-12, n = 19) in stage random of the estrous cycle received the insertion of CIDR (D0). On the day of implant removal (D12) was administered intramuscularly, 0.075 mg cloprostenol (Veteglan®, Hertape Calier, Brazil) and 300 IU equine chorionic gonadotropin (eCG, Novormon®, MSD Animal Health, Brazil). The group 6 (G-6, n= 24) received the same protocol of G-12, however the permanence of CIDR was 6 days. Approximately 50 hours after CIDR removal, both groups were inseminated at fixed time by artificial insemination by laparoscopy with frozen semen containing 200x106 sperm of a single player of Dorper. Approximately 40 days after the artificial insemination pregnancy diagnosis was performed using ultrasound device. Blood samples were taken every 4 hours from 10 animals per group, to measure LH and progesterone plasma concentrations by radioimmunoassay (RIA). From the 20 animals used for progesterone plasma concentration, the percentage of animals showing LH surge in G-6 was 70% (7/10) and the G-12 was 90% (9/10). The maximum amplitude of the LH peak did not differ between the G-6 and G-12 groups (P> 0.05) and the lengh of LH peak was 16,60±2,76 h in the G-6 and 16,40±2,40 h in G-12 (P>0,05). The interval between the implant removal and the LH surge was (P >0.05) in the G-6 27,42±2,76 h and 28,88±5,20 h in G-12. The overall pregnancy rate per group was not different (G-6 58,33% e G-12 52,63%). It was concluded that the permanence of progesterone (6 vs 12 days) did not alter the characteristics of the LH peak in sheep (St. Agnes x Texel) in the tropics, out of the breeding season. / A hipótese testada pelo presente trabalho foi que o tempo de permanência do implante de progesterona (curto vs longo), em protocolo de IATF, altera as características do pico de LH. Desta forma, o objetivo do presente estudo foi determinar o momento, duração e amplitude do pico de LH em ovelhas (Texel x Santa Inês) utilizando protocolos de IATF de curta (6 dias) ou longa duração (12 dias) em ovelhas ciclando na contra estação reprodutiva. Foram utilizadas 43 ovelhas (Texel-Te x Santa Inês-SI), divididas em dois grupos de acordo com o tempo de permanência do dispositivo vaginal de liberação de progesterona de 1° uso (Easy-Breed CIDR®, Pfizer, Brasil). No grupo 12 dias (G-12, n=19), em estádio aleatório do ciclo estral receberam a inserção do CIDR (D0). No dia da retirada do implante (D12) foram administrados, por via intramuscular, 0,075 mg de cloprostenol (Veteglan®, Hertape Calier, Brasil) e 300 UI de gonadotrofina coriônica equina (eCG, Novormon®, MSD Saúde Animal, Brasil). O grupo 6 (G-6, n=24) recebeu o mesmo protocolo do G-12, porém a permanência do CIDR foi de 6 dias. Aproximadamente 50 horas após a retirada do CIDR as ovelhas, de ambos os grupos, foram inseminadas em tempo fixo por laparoscopia com sêmen congelado contendo 200x106 espermatozóides, de um único reprodutor, da raça Dorper. Aproximadamente 40 dias após as inseminações artificiais foi realizado o diagnóstico de prenhez utilizando aparelho de ultra-som. Foram realizadas colheitas de sangue, de 4 em 4 horas em 10 animais por grupo, para mensuração das concentrações plasmáticas de LH e progesterona por radioimunoensaio (RIA). Dos 20 animais utilizados para mensuração das concentrações plasmáticas, a porcentagem de animais que apresentaram pico de LH no G-6 foi de 70% (7/10) e no G-12 foi 90% (9/10). A amplitude máxima do pico de LH não diferiu entre os grupos G-6 e G-12 (P>0,05) e a duração do pico de LH foi 16,60±2,76 h no grupo G-6 e 16,40±2,40 h no grupo G-12 (P>0,05). O intervalo entre a retirada do implante e o pico de LH foi (P>0,05) ocorreu 27,42±2,76 h no grupo G-6 e 28,88±5,20 h no G-12. A percentagem de prenhez total por grupo não diferiu (G-6 58,33% e G-12 52,63%). Conclui-se que o tempo de permanência da progesterona (6 vs 12 dias) não alterou as características do pico de LH em ovelhas (Texel x Santa Inês) nos trópicos, na contra estação reprodutiva.
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