Les études sur les couples traités par fécondation in vitro (FIV) ont jusqu’à présent porté essentiellement sur l’évaluation du succès en FIV. Très peu de données sont disponibles sur le devenir à long terme de couples traités par FIV. L’objectif de ce travail était d’estimer la fréquence de réalisation du projet parental à long terme, et d’étudier les facteurs associés aux interruptions précoces des traitements et aux naissances naturelles.L’enquête DAIFI-2009 a inclus 6 507 couples ayant débuté un programme de FIV en 2000-2002 dans l’un des 8 centres de FIV participant à l’étude. Les données médicales des couples et leur parcours dans le centre ont été obtenus à partir des dossiers médicaux des centres de FIV pour tous les couples. L’information sur le devenir des couples après le départ du centre a été obtenue par questionnaire postal auprès des couples en 2008-2009 (38% de participation 7 à 9 ans après l’initiation des FIV). L’étude des facteurs associés à la participation à l’enquête postale suggérait que la fréquence de réalisation du projet parental estimée sur les répondants seulement pourrait être biaisée. Les différentes méthodes mises en œuvre pour corriger la non réponse (pondération, imputation multiple) n’ont pas modifié l’estimation de la fréquence de réalisation du projet parental. Au total, 7 à 9 ans après l’initiation des FIV, 60% des couples ont réalisé leur projet parental de façon biologique, suite à un traitement ou suite à une conception naturelle. Lorsque les adoptions sont aussi prises en compte, 71% des couples ont réalisé leur projet parental. Après l’échec d’une première tentative de FIV, un couple sur 4 (26%) a interrompu les FIV dans le centre d’inclusion. Globalement, les couples avec de mauvais facteurs pronostiques ont un plus grand risque d’interrompre les FIV. Cependant, la proportion plus importante d’interruption parmi les couples avec une origine inexpliquée de l’infécondité pourrait s’expliquer par la survenue plus fréquente de naissance naturelle dans ce sous-groupe de couples. Parmi les couples n’ayant pas eu d’enfant suite aux traitements, 24% ont ensuite conçu naturellement en médiane 28 mois après l’initiation des FIV. Parmi les couples ayant eu un enfant suite aux traitements, 17% ont ensuite conçu naturellement en médiane 33 mois après la naissance de l’enfant conçu par AMP. Les facteurs associés aux naissances naturelles sont des indicateurs d’un meilleur pronostic de fertilité, particulièrement chez les couples sans enfant AMP.L’enquête DAIFI-2009 a permis d’apporter des informations sur le parcours à long terme des couples traités par FIV qui n’avait jusqu’à présent été que peu étudié, souvent sur de faibles effectifs et avec un suivi plus court. Ces résultats doivent apporter de l’espoir aux couples inféconds, puisque la majorité d’entre eux ont finalement réalisé leur projet parental, même si cela peut prendre de nombreuses années. / Until now, most studies of couples treated by in vitro fertilization (IVF) have been centered on IVF success. Very few data are available on the long-term outcome of these couples, including spontaneous conception and adoptions. This work aimed to estimate the long-term cumulative parenthood rate, and to study factors associated with early IVF discontinuation and with spontaneous live births.The DAIFI study is a retrospective cohort including 6,507 couples who began IVF in 2000-2002 in one of the eight participating French IVF centres. Medical data on all couples were obtained from centre databases. Information on long-term outcome after leaving the IVF center was collected by postal questionnaire sent to couples in 2008-2010 (7 to 9 years after IVF initiation, participation rate 38%). Study of factors associated with participation in the postal survey suggested that the cumulative parenthood rate estimated only in participants might be biased. The different methods used to correct for non-response bias (inverse probability weighting, multiple imputation) did not modify the estimation of the cumulative parenthood rate obtained with the complete case approach. Finally, 7 to 9 years after IVF initiation, the cumulative parenthood rate was estimated at 60%, including live births following IVF, other treatment or spontaneous conception. When adoptions were also considered, the cumulative parenthood rate reached 71%. After a first failed IVF cycle, just over one couple out of four (26%) discontinued IVF treatment. Globally, couples with poor prognostic factors had a higher risk of early discontinuation of IVF treatment. However, the higher proportion of early discontinuation observed among couples with unexplained infertility could be linked to a higher chance of spontaneous pregnancy in this subpopulation. Among couples who remained childless after treatment, 24% later had a spontaneous live birth (SLB), at a median of 28 months after the first IVF attempt. Among couples who had had a child during medical treatment, 17% later had an SLB, at a median of 33 months after the birth following medical treatment. Regarding factors associated with SLB, they can be viewed as indicators of a better fertility prognosis, especially among unsuccessfully treated couples.The DAIFI study has provided information on the long-term outcome of couples treated by IVF, which has until now been little studied, often on small samples and with a shorter duration of follow-up. These results should give hope to infertile couples as nearly three couples out of four finally became parents, even if it may take many years.

La ICSI porcina es una herramienta con gran potencial aplicativo en diversos campos, entre los que destacan la producción de animales transgénicos, y la recuperación de razas en peligro de extinción. Aunque en la actualidad existen referencias de obtención de descendencia viva, el rendimiento es inferior al de otras especies, posiblemente debido al desconocimiento de las condiciones idóneas, y la dificultad de los cigotos para alcanzar el estadío de blastocisto in vitro. El presente trabajo se llevó a cabo para determinar diferentes factores que podrían afectar al rendimiento de la técnica, estudiando el efecto de 1) la secuencia de cultivo de los zigotos recién inyectados; 2) modificaciones en el sistema de MIV tradicional, y por último 3) la activación exógena del ovocito mediante la inyección de inositol trifosfato con el espermatozoide. El objetivo global de este estudio fue el de incrementar el rendimiento final de la ICSI en la especie porcina. / ICSI in pigs is a tool with an important applicable potential in diverse fields. One of this is the production of transgenic animals, and the conservation of endangered species. Even though there are some cases of living offspring, its output is still quite low comparing to other species, possibly due to unknown factors referring to ideal conditions for the development, and to the difficulty of the zygotes to reach the blastocyst stage in vitro. The goal of this study was to evaluate different factors affecting the ICSI performance. This was done by studying 1) the sequence of culture of the injected oocytes; 2) In vitro maturation (IVM) modifications, through meiotic inhibitors, such as roscovitine, and changes in IVM duration time, and finally 3) the exogenous oocyte activation through inositol triphosphate (InsP3) injection together with the sperm. The main objective of this study was to increase the final performance of ICSI in pigs.

Les études sur les couples traités par fécondation in vitro (FIV) ont jusqu'à présent porté essentiellement sur l'évaluation du succès en FIV. Très peu de données sont disponibles sur le devenir à long terme de couples traités par FIV. L'objectif de ce travail était d'estimer la fréquence de réalisation du projet parental à long terme, et d'étudier les facteurs associés aux interruptions précoces des traitements et aux naissances naturelles.L'enquête DAIFI-2009 a inclus 6 507 couples ayant débuté un programme de FIV en 2000-2002 dans l'un des 8 centres de FIV participant à l'étude. Les données médicales des couples et leur parcours dans le centre ont été obtenus à partir des dossiers médicaux des centres de FIV pour tous les couples. L'information sur le devenir des couples après le départ du centre a été obtenue par questionnaire postal auprès des couples en 2008-2009 (38% de participation 7 à 9 ans après l'initiation des FIV). L'étude des facteurs associés à la participation à l'enquête postale suggérait que la fréquence de réalisation du projet parental estimée sur les répondants seulement pourrait être biaisée. Les différentes méthodes mises en œuvre pour corriger la non réponse (pondération, imputation multiple) n'ont pas modifié l'estimation de la fréquence de réalisation du projet parental. Au total, 7 à 9 ans après l'initiation des FIV, 60% des couples ont réalisé leur projet parental de façon biologique, suite à un traitement ou suite à une conception naturelle. Lorsque les adoptions sont aussi prises en compte, 71% des couples ont réalisé leur projet parental. Après l'échec d'une première tentative de FIV, un couple sur 4 (26%) a interrompu les FIV dans le centre d'inclusion. Globalement, les couples avec de mauvais facteurs pronostiques ont un plus grand risque d'interrompre les FIV. Cependant, la proportion plus importante d'interruption parmi les couples avec une origine inexpliquée de l'infécondité pourrait s'expliquer par la survenue plus fréquente de naissance naturelle dans ce sous-groupe de couples. Parmi les couples n'ayant pas eu d'enfant suite aux traitements, 24% ont ensuite conçu naturellement en médiane 28 mois après l'initiation des FIV. Parmi les couples ayant eu un enfant suite aux traitements, 17% ont ensuite conçu naturellement en médiane 33 mois après la naissance de l'enfant conçu par AMP. Les facteurs associés aux naissances naturelles sont des indicateurs d'un meilleur pronostic de fertilité, particulièrement chez les couples sans enfant AMP.L'enquête DAIFI-2009 a permis d'apporter des informations sur le parcours à long terme des couples traités par FIV qui n'avait jusqu'à présent été que peu étudié, souvent sur de faibles effectifs et avec un suivi plus court. Ces résultats doivent apporter de l'espoir aux couples inféconds, puisque la majorité d'entre eux ont finalement réalisé leur projet parental, même si cela peut prendre de nombreuses années.

Abstract Feline immunodeficiency virus (FIV) is an important infectious agent of domestic cats worldwide. It has been classified into the Lentivirus genus of the Retroviridae family, together with human immunodeficiency virus (HIV). Five FIV subtypes (A, B, C, D and E) have been described based on sequence variation of the V3-V5 region of the envelope (env) gene. There is considerable sequence diversity within and between subtypes, which has been a major obstacle in the development of a successful vaccine. However, an FIV vaccine that incorporates inactivated whole viruses from subtypes A and D is now commercially available. Although the vaccine has been shown to be efficacious in protecting against challenge with homologous and a heterologous (subtype B) subtypes, its effectiveness against other viral variants is unknown. Therefore, identifying the type and diversity of FIV strains in different regions is important to establish the potential efficacy of the vaccine in areas where vaccination is to be implemented. The proviral DNA sequence of the V3-V5 region of the env gene was determined for 102 FIV-infected cats from locations in Australia, New Zealand and South Africa. Subtype A was the predominant subtype in Australia and South Africa, although subtype B and C were also identified in each of these countries, respectively. Both subtypes A and C were also present in New Zealand. Of interest, there were some samples in New Zealand and South Africa that demonstrated subtype assignment discrepancies when different regions of the genome were analysed, suggesting co-infection and/or recombination. Cats infected with FIV exhibit varying degrees of immunological impairment. Currently, prognosis for an FIV-infected cat is based on clinical signs alone, which is a relatively subjective measure. In HIV-infected patients it is recognised that viral RNA load correlates with disease stage and prognosis. This PhD research tested whether viral RNA load may be a useful prognostic marker in FIV infection. A real-time PCR assay was developed to quantify plasma viral RNA load in 42 FIV-infected cats at three different clinical stages (1:healthy, 2:unwell without signs of immunodeficiency, 3:unwell with signs of immunodeficiency). In cats older than 5 years of age, log-transformed viral RNA loads were significantly higher in cats in category 3 compared to cats in category 1. There were no significant differences in the viral RNA load of older cats in category 2 compared to category 1. There were no cats younger than 5 years of age in category 3 and there was no significant difference in viral RNA load between young cats in categories 1 and 2. Of the 15 cats for which follow-up data was available, eight showed no change in clinical signs, and seven showed a worsening of clinical signs with six of these showing a progression of clinical category including death. One of the cats in category 2 that progressed clinically had one of the highest viral RNA loads of cats in that category. Three of four cats from category 3 that were followed had either died or been euthanised. Two of these cats had among the highest viral RNA loads in the whole study, while the remaining cat (for which the definitive cause of death was not confirmed) had a relatively low viral RNA load. In summary, measurement of viral RNA load was found to be a potentially useful clinical and prognostic marker but further work is required to better assess its usefulness to veterinarians. Serum acute phase proteins were investigated as possible candidate markers of FIV disease with the aim of developing a more simplified assay that could be used as a prognostic marker for FIV infection. Blood samples from 43 FIV-infected and 25 FIV-negative cats were assayed for the concentration of four acute phase proteins. Both healthy and sick cats were included in the study. Compared to healthy cats, sick cats had significantly higher concentrations of serum amyloid A (P<0.05). Alpha 1-acid glycoprotein and haptoglobin were also found to be in higher concentrations in sick cats (P<0.1). Other variables such as age and gender were also associated with acute phase protein concentrations. With respect to FIV infection, it was found that in sick cats, serum amyloid A, in combination with the age of the cat, was the best predictor of FIV viral RNA load. Alpha 1-acid glycoprotein and haptoglobin were not significantly associated with FIV viral RNA load. Although health status did not influence albumin levels, they were found to be significantly lower in FIV-positive cats in comparison to FIV-negative cats (P<0.05). The frequent monitoring of viral RNA loads and CD4+ lymphocyte counts that is performed on HIV-infected patients is cost prohibitive in veterinary patients. This study showed that there is potential for the use of acute phase protein concentrations (in particular serum amyloid A) as alternative prognostic tools in FIV-infected cats. Further work, particularly longitudinal studies, is required to more definitively define changes in viral RNA load and acute phase protein concentrations throughout the course of FIV infection.

Made available in DSpace on 2016-12-08T16:24:17Z (GMT). No. of bitstreams: 1 PGCA14MA122.pdf: 692452 bytes, checksum: e25e927c8d8beb4236a0bb259e17524a (MD5) Previous issue date: 2014-02-21 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / Oxidative stress and premature sperm capacitation, generated during cryopreservation of ram semen, reduces their viability, especially after cervical insemination. The use of seminal plasma (SP) and negative pressure have produced protection and the reversion of such damages. Two experiments evaluated these potential enhancers of cryotolerance, and a third experiment compared 2 methods of cervical AI. In experiment 1 ram semen was subjected to the treatments: (TC) control or negative pressure of 200mBar (P200); 500mBar (P500) and 800mBar (P800). In experiment 2, the PS from rams, stallions and bulls was lyophilized (L) and its protein measured. From each SP 600&#956;g of protein per mL was aded to the freezing diluent used, compounding the experimental groups: control (TC), ovine PS (PSLO), bovine PS (PSLB) and equine PS (PSLE). Experiment 3 evaluated 2 methods of AI, the superficial cervical AI (G1), and deep intrauterine or cervical AI with clamping the vaginal fornix (G2). The in vitro data were subjected to ANOVA and test T, and the pregnancy rate to the chi square test, all with 5% significance level. In the experiment 1 higher progressive motility (PM) was observed in TC (49%) compared to P200 (40.9%), P500 (38.9%) and P800 (38.9%) treatments. In PM during the test the thermal resistance (TTR), MP after percoll (PP), acrosome integrity (IAC), IAPP and membrane integrity (MI), there was no difference between the groups. In cleavage rate P800 (34.5%) was less than P200 (51.2%) and P500 (50.9%) did not differ from the control (44.3%). In conclusion the P500 is the most appropriate for use in ram semen cryopreservation, enabling high rates of cleavage after heterologous IVF, maintain membrane integrity. Experiment 2 evaluated MP, MPPP and cleavage rate after heterologous IVF in all groups, with the best group compared with the control in: CASA system; acrossoma integrity (FITC-PSA), membrane stability (M540), chromatin integrity (acridine orange), apoptosis (annexin) and potential of mitochondria (Mitotracker). Also the PSLE showed higher cleavage rate (71.37%), indicating a greater ability to oocyte penetration. The PSLE showed higher VCL (PC-163.5&#956;m/s, PSLE-186.2&#956;m/s) and ALH (PC9&#956;m PSLE-8.2&#956;m) in CASA evaluation, compared to control. In flow cytometry the annexin test revealed a greater amount of non-apoptotic viable cells in PSLE (38.9%), compared to TC (32.1%). In experiment 3 there was no difference in pregnancy rates after superficial (33.3%) or deep and intrauterine (52.2%) IA, possibly due to the reduced number of animals used / O estresse oxidativo e a precoce capacitação espermática, gerados na criopreservação do sêmen ovino, reduzem sua viabilidade, principalmente na inseminação cervical. O uso de plasma seminal (PS) e a pressão negativa têm produzido a proteção e reversão desses danos. Dois experimentos avaliaram esses potenciais melhoradores da criotolerância, e um terceiro avaliou dois métodos de IA cervical. No experimento 1 o sêmen ovino foi submetido aos tratamentos: controle (TC), pressão de 200mBar (P200); 500mBar (P500) e 800mBar (P800). No experimento 2 o PS de carneiros, garanhões e touros foi liofilizado (L) e sua proteína dosada. De cada PS, o equivalente a 600&#956;g de proteína por mL, foi adicionado ao diluente de congelamento, compondo os grupos experimentais: controle (TC), PS ovino (PSLO), PS bovino (PSLB) e PS equino (PSLE). O experimento 3 avaliou 2 métodos de IA, a cervical superficial (G1) e a cervical profunda com pinçamento do fundo de saco vaginal (G2). Os dados in vitro foram submetidos a análise de variância e teste T, e a taxa de prenhez ao chi-quadrado, todos com significância de 5%. No experimento 1, maior motilidade progressiva (MP) foi observada no TC (49%) frente aos tratamentos P200 (40,9%), P500 (38,9%) e P800 (38,9%). Na MP durante o teste de termo resistência (TTR), MP após percoll (PP), integridade de acrossoma (IAC), IACPP, integridade de membrana (IM) e IMPP, não houve diferença entre os grupos. Na clivagem P800 (34,5%) foi inferior a P200 (51,2%) e P500 (50,9%), não diferindo do controle (44,3%). Conclui-se que a P500 é a mais adequada para uso com sêmen ovino, não reduzindo a viabilidade após o congelamento e proporcionando elevada taxa de clivagem após FIV heteróloga. O experimento 2 avaliou MP, MPPP e clivagem após FIV heteróloga de todos os grupos, sendo o melhor grupo comparado ao controle através de: sistema CASA, integridade de acrossoma (FITC-PSA), estabilidade de membrana (M540); integridade de cromatina (acridina orange); apoptose (anexina) e potencial de mitocôndria (mitotracker). O PSLE apresentou a maior taxa de clivagem (71,37%), evidenciando sua maior capacidade de penetração nos oócitos. Observou-se superioridade do PSLE nosparâmetros VCL (PC-163,5&#956;m/s, PSLE-186,2&#956;m/s) e ALH (PC-9&#956;m, PSLE- 8,2&#956;m) do CASA, em relação ao controle. Na citometria de fluxo, o teste da anexina revelou maior quantidade de células viáveis não apoptóticas com o PSLE (38,9%) em relação ao TC (32,1%). No experimento 3 não houve diferença na prenhez após IA superficial (33,3%) e profunda (G2 52,2%), possivelmente devido ao número reduzido de animais

Conselho Nacional de Desenvolvimento Científico e Tecnológico / Preimplantation genetic diagnosis (PGD) is becoming a current issue in animal reproduction biotechnology due to economical reasons. Predetermining the sex of offspring is one example of PGD. This study aimed to determine the percentage of male and female bovine embryos in vitro produced after oocyte fertilization with Percoll density gradient centrifugation or with self-migration (swim-up) selected semen. In experiment 1, sperm selection was performed by 90%-45% discontinuous Percoll density gradient centrifugation (T1) and swim-up (T2). In experiment 2, along side the discontinuous gradient, a 67.5% continuous density gradient, and centrifugation time of 5 and 10 minutes were used. A total of 4 treatment groups was defined (TI = continuous, 5 minutes, TII = discontinuous, 5 minutes, TIII = continuous, 10 minutes and TIV = discontinuous, 10 minutes). Polymerase chain reaction (PCR) was used to determine the sex of the embryos. T1 (n=185) resulted in 48.65% (n=90) male embryos and 51.35% (n=95) female embryos and T2 (n=142) in 58.45% (n=83) male and 41.55% (n=59) female embryos. In experiment 2, the percentages of male and female embryos obtained in TI (n=93), TII (n=70), TIII (n=82) and TIV (n=82) were 49.46% (n=46) and 50.54% (n=47), 57.14% (n=40) and 42.86% (n=30), 36.59% (n=30) and 63.41% (n=52) and 48.78% (n=40) and 51.22% (n=42), respectively. There was no difference on the percentage of males and females in all treatment groups from experiments 1 and 2 when these were individually compared to the expected percentage of 50% of each sex. There was also no difference in male and female embryo percentage between treatment groups in experiments 1 and 2. / O diagnóstico genético pré-implantação (DGP) vem se destacando na área da biotecnologia da reprodução animal por motivos econômicos. Um exemplo de DGP é a predeterminação do sexo da prole. Neste estudo foi verificada a percentagem de embriões bovinos machos e fêmeas produzidos in vitro após a fertilização de oócitos com sêmen selecionado por centrifugação em gradiente de densidade de Percoll ou por migração ascendente (swim-up). No experimento 1 a seleção espermática foi realizada usando o gradiente descontínuo de Percoll de 90% e 45% (T1) e o swimup (T2). No experimento 2 foi utilizado, além do gradiente descontínuo, um gradiente contínuo de densidade de Percoll de 67,5%, e tempos de centrifugação de 5 e 10 minutos, totalizando 4 tratamentos (TI = contínuo 5 minutos, TII = descontínuo 5 minutos, TIII = contínuo 10 minutos e TIV = descontínuo 10 minutos). A sexagem dos embriões foi realizada através da técnica da reação em cadeia da polimerase (PCR). No T1 (n=185) foram obtidos 48,65% (n=90) de embriões masculinos e 51,35% (n=95) de femininos e no T2 (n=142) 58,45% (n=83) foram machos e 41,55% (n=59) fêmeas. No experimento 2, a percentagem de embriões masculinos e femininos no TI (n=93), TII (n=70), TIII (n=82) e TIV (n=82) foi de 49,46% (n=46) e 50,54% (n=47), 57,14% (n=40) e 42,86% (n=30), 36,59% (n=30) e 63,41% (n=52), e 48,78% (n=40) e 51,22% (n=42), respectivamente. Não houve alteração na percentagem de machos e fêmeas nos tratamentos dos experimentos 1 e 2 quando estes tratamentos foram comparados individualmente com a percentagem teoricamente esperada de 50% de cada sexo. Também não houve alteração na percentagem de machos e fêmeas na comparação entre os dois tratamentos do experimento 1 e entre os quatro tratamentos do experimento 2.

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.

Justificativa: Falhas recorrentes de implantação (FRI) são uma fonte de grande frustração para pacientes e especialistas que, frequentemente buscam intervenções com o objetivo de atingir resultados favoráveis. A prevalência exata dessa condição é de difícil estimativa, uma vez que existem diversas definições para caracterizá-la. Diversas intervenções que visam a melhora dos resultados reprodutivos dessas pacientes já foram propostas. Entretanto, nenhuma revisão sistematizada abordou, simultaneamente, todas as potenciais estratégias para esse grupo de mulheres. Dessa forma, a eficácia e a segurança dessas intervenções não são bem definidas. Devido ao alto número de intervenções descritas para esse fim, julgamos que uma metanálise que as contemple de forma abrangente é importante para casais, especialistas e pesquisadores do assunto. Objetivos: Avaliar a eficácia e segurança das intervenções que visam a melhora do resultado reprodutivo das pacientes com FRI submetidas a um novo tratamento de reprodução assistida. Métodos de busca: As buscas por estudos randomizados e controlados, publicados e em andamento, foram realizadas nas principais bases de dados eletrônicas. Adicionalmente, as listas de referências de estudos incluídos e revisões semelhantes foram avaliadas pelos autores. A última busca eletrônica foi realizada em fevereiro de 2018. Critérios de Elegibilidade: Foram considerados elegíveis apenas os estudos verdadeiramente randomizados que comparassem quaisquer intervenções destinadas a esse grupo depacientes. No presente estudo, consideramos como FRI a história de duas ou mais falhas prévias. Extração e análise de dados: Dois autores realizaram, individualmente, a seleção de estudos, extração de dados e análise do risco de viés. Discordâncias foram resolvidas em consulta a um terceiro autor. Os pesquisadores de estudos potencialmente elegíveis foram contatados sempre que necessário para obtenção de informações adicionais. Resultados: Foram identificados 2794 registros; desses, 62 estudos foram incluídos, representando uma população de 9308 pacientes, submetidas a 26 intervenções diferentes. Informações sobre 24 intervenções e 8461 pacientes foram submetidas à análise quantitativa. Não há evidências de alta ou moderada qualidade de que alguma dessas intervenções seja realmente eficaz para a melhora dos resultados reprodutivos de pacientes com FRI. As seguintes intervenções se mostraram benéficas: assisted hatching, injúria endometrial, histeroscopia, uso de FSH urinário + recombinante para estimulação endometrial e administração intrauterina de hCG. Porém as evidências são de baixa qualidade, o que nos traz incerteza em relação aos seus reais efeitos. Todas as outras intervenções identificadas não permitiram nenhuma outra conclusão adicional, uma vez que as evidências foram avaliadas como de muito baixa qualidade ou não foram encontrados estudos randomizados que as tivessem avaliado. Conclusões: Evidências de ensaios clínicos randomizados não sustentam o uso de nenhuma intervenção para a melhora dos resultados reprodutivos de pacientes com FRI. Todos os achados foram julgados como de baixa ou muito baixa qualidade, o que nos traz incerteza quanto aos seus reais efeitos na prática clínica. Deve-se estar ciente de que o emprego de tais intervenções impõe despesas e riscos adicionais para as pacientes. Além disso, a falta de critérios universalmente aceitos para odiagnóstico de falha recorrente de implantação é uma importante limitação para o avanço do conhecimento sobre essa condição / Background: Recurrent implantation failure (RIF) is a source of deep frustration to couples and clinicians, who often look for interventions to improve the reproductive outcomes. Its exact prevalence is difficult to determine because there are several definitions used to describe the condition. Several interventions aiming to improve reproductive outcomes for such patients have been studied. However, there are no systematic reviews that focus on all potential interventions for improving reproductive outcomes in women with RIF undergoing assisted reproduction techniques. The efficacy and safety of these interventions are not clear. Because of the large number of potential interventions for this condition, it would be very difficult to be aware of the current evidence for all of them. We believe this systematic review is important for subfertile couples, clinicians and researchers. Objectives: To assess the efficacy and safety of interventions designed to improve reproductive outcomes in women with RIF undergoing ART. Search methods: We searched for randomised controlled trials (RCT) in electronic databases (Cochrane Gynaecology and Fertility Group (CGF), The Cochrane Central Register of Controlled Trials, MEDLINE Ovid, EMBASE Ovid, PsycINFO, PsycINFO Ovid, CINAHL, LILACS), trials registers (ClinicalTrials.gov, ISRCTN registry, The WHO International Clinical Trials Registry Platform, World Health Organization International Clinical Trials Registry Platform) and grey literature (OpenGrey); in addition, we handsearched the reference lists of included studies and similar reviews. We performed the last electronic search on 22 Feb 2018.Selection criteria: We considered eligible only truly randomised controlled trials comparing any intervention designed to improve outcomes in women with repeat implantation failure (RIF) compared to other intervention, placebo or no treatment. For study selection, we defined RIF as two or more previous failures Data collection and analysis: Two authors independently performed study selection, data extraction, and assessment of the risk of bias. Any disagreements were solved by consulting a third review author. Study\'s authors were contacted whenever needed to solve any queries. Results: the search retrieved 2794 records; from those, sixty-two studies were included, comprising 9308 participants, submitted to 26 different interventions. Data from 24 interventions and 8461 participants were pooled for quantitative analysis. We found no high or even moderate quality evidence that any of the tested interventions are really effective to improve reproductive outcomes of women with RIF undergoing a new IVF treatment. We observed low-quality evidence of benefit for women with RIF with the following interventions: assisted hatching, endometrial Injury, hysteroscopy, the use of human + recombinant FSH for ovarian stimulation and intrauterine hCG administration. All the other listed interventions did not allow any further conclusion: either very low-quality evidence or no evidence from RCTs. Conclusions: Evidence from RCTs does not support the use of any specific intervention for improving reproductive outcomes in women with RIF. All evidences were deemed of low to very low quality, which makes us uncertain of their real effectiveness on clinical practice. One should be aware that the employment of such interventions imposes additional expenses and risks. Additionally, the lack of universally accepted criteria for recurrent implantation failure is an important limitation for the advance of knowledge regarding this condition.More studies are needed to evaluate their real effect. Maybe even more importantly is to create universally accepted criteria for defining implantation failure; only them one will be able to test interventions for this specific group

A vibração induzida por escoamento é parte intrínseca do transporte de fluidos. Por exemplo, na indústria de petróleo e gás esse fenômeno pode ser encontrado em tubulações, tanto no setor upstream, quando downstream. Essas vibrações são produto das forças geradas pelo escoamento e, portanto, carregam informações sobre sua fenomenologia. No caso de escoamento bifásico em tubo, resultados experimentais indicam forte influência da velocidade da mistura, fração de vazio e padrão de escoamento no comportamento dinâmico da estrutura. Contudo, pouco foi feito na tentativa de obter informações do escoamento a partir da reposta estrutural. Assim, o objetivo do presente estudo é desenvolver métodos para a previsão dos parâmetros do escoamento baseados na resposta de um tubo submetido a escoamento bifásico. Foi conduzido um trabalho experimental da vibração induzida por diversos padrões gás-líquido numa tubulação horizontal (PVC &#216;3/4\'\') duplamente engastada, com água e ar como fluidos de trabalho. A partir de uma abordagem analítica, corroborada com resultados experimentais para escoamento monofásico e bifásico, estabelece-se a existência de uma relação, de natureza quadrática, entre a velocidade de atrito e o desvio padrão da aceleração. Dado que a velocidade de atrito é função do fator de atrito bifásico, um método para a sua previsão é desenvolvido. Ele prevê de maneira precisa os dados coletados; todos eles com erro percentual menor do que 30%. O método foi comparado também com dados experimentais e modelos da literatura, mostrando boa concordância. Além disso, apresenta-se uma relação entre a frequência pico da resposta e a fração de vazio homogênea. No fim, são apresentados: (i) um método de identificação de escoamento pistonado, baseado na superposição dos mecanismos de vibração por turbulência e intermitente, com desempenho mínimo de 81.8%; (ii) um método experimental para determinação da velocidade da mistura (J) e fração de vazio homogênea (&beta;). Os melhores resultados são obtidos para os padrões disperso e pistonado, prevendo adequadamente os parâmetros J e &beta; com erro percentual absoluto médio de 24.1% e 20.65%, respectivamente. / Flow-induced vibration is intrinsic to piping problems. For example, in the oil and gas industry the FIV phenomenon can be found in pipe flow both in upstream and downstream applications. The structural vibration response contains information about the flow phenomenology. In the case of two-phase pipe flow, experimental results show a strong influence of mixture velocity, void fraction and flow pattern on pipe structural dynamics. However, efforts to obtain information of the flow from pipe response have been scanty. The goal of this study is to develop two-phase flow parameters predictive methods based on the structural pipe response. An experimental study of flow-induced vibration was carried out for several flow patterns in a clamp-clamp straight pipe (PVC &#216;3/4\'\'), with air and water as working fluids. From an analytical approach, a quadratic relationship between shear velocity and standard deviation of acceleration is proposed and validated against the experimental data of single and two-phase flow. Since the shear velocity depends on the friction factor, a method to predict two-phase friction factor is presented. The method predicts accurately our experimental data with a mean absolute error up to 30%. Good agreement was also found when it was compared with some models and experimental data from the literature. Furthermore, an expression to correlate peak frequency and homogeneous void fraction as a function of added mass is offered. Finally, we present: (i) a slug flow identification technique based on the superposition of the turbulence and intermittent flow-induced vibration mechanisms, with performance of 81.8% and (ii) an experimental methodology to estimate mixture velocity (J) and homogeneous void fraction (&beta;). The latter method shows better agreement for dispersed and slug flow-patterns, predicting J and &beta; with a mean absolute error of 24.1% e 20.65%, respectively.

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.