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1.
Mol Reprod Dev ; 86(2): 145-155, 2019 02.
Artigo em Inglês | MEDLINE | ID: mdl-30418697

RESUMO

For the present study we asked whether the endometrial fluid lipidomic may be a useful approach to predict endometrial receptivity in freeze-all cycles. For this case-control study, endometrial fluid samples were collected from 41 patients undergoing freeze-all cycles. Samples were split depending on the pregnancy outcome: positive group (n = 24) and negative group (n = 17). Data were acquired by the matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. Principal component analysis (PCA) and partial least squares discriminant analysis (PLS-DA) were applied. A list of potential biomarker ion ratios was obtained and the values were used to build a receiver operating characteristic (ROC) curve to predict pregnancy success. The lipid categories were attributed by LIPID MAPS database. Ion ratios were established according to their correlations and used for the analysis. The PCA showed a tendency of separation between the studied groups, whereas the PLS-DA was able to clearly distinguish them. Fifteen ratios (13 hyper-represented in the negative and two hyper-represented in the positive group) were selected according to their importance for model prediction. These ratios were used to build the ROC curve, which presented an area under curve of 84.0% (95%CI: 69.2-97.4%; p = 0.009). These findings suggest that lipidomic profiling of endometrial fluid may be a valuable tool for identifying the time interval comprising the window of implantation.


Assuntos
Bases de Dados Factuais , Implantação do Embrião , Endométrio/metabolismo , Metabolismo dos Lipídeos , Modelos Biológicos , Adulto , Biomarcadores/metabolismo , Líquidos Corporais/metabolismo , Feminino , Humanos , Gravidez
2.
J Assist Reprod Genet ; 36(3): 461-471, 2019 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-30515618

RESUMO

PURPOSE: To study the association between body mass index (BMI), waist circumference (WC), waist-to-hip ratio (WHR), and waist-to-height ratio (WHtR), and response to controlled ovarian stimulation, semen quality, and intracytoplasmic sperm injection (ICSI) outcomes. METHODS: This prospective observational study included data from 402 couples undergoing ICSI between January/2010-September/2016. Participants had their weight, height, waist, and hip measured before ICSI treatment. RESULTS: Gonadotropin dose was positively associated with maternal WHR. Maternal WC and WHR negatively influenced the estradiol level, number of follicles, oocytes, and mature oocytes. Maternal BMI negatively influenced oocyte yield. Semen volume was negatively influenced by male BMI. Sperm concentration was negatively associated with WHR and WHtR. Progressive sperm motility was negatively influenced by all anthropometric measures (AMs). The TMSC was negatively affected by all AM but WC. Significant differences were observed in all ICSI outcomes except pregnancy and miscarriage rates, favoring women with AM below the cut off values compared to women above the cut off. It was observed that men with AM below the cut off values showed significantly higher rates of fertilization, embryo and blastocyst development, implantation, and pregnancy than men above the cut off. When combined maternal and paternal AM were analyzed, significant differences were observed in fertilization, embryo and blastocyst development and implantation rates, favoring couples with normal BMI, WC, and WHR. CONCLUSIONS: Lower AMs have a protective effect on fertility. Awareness and counseling on how to minimize its impact in both partners could improve ICSI outcomes.


Assuntos
Implantação do Embrião/fisiologia , Fertilização in vitro/métodos , Oócitos/crescimento & desenvolvimento , Injeções de Esperma Intracitoplásmicas , Adulto , Antropometria/métodos , Índice de Massa Corporal , Feminino , Humanos , Masculino , Oócitos/metabolismo , Pais , Gravidez , Taxa de Gravidez , Análise do Sêmen/métodos , Contagem de Espermatozoides , Motilidade dos Espermatozoides , Relação Cintura-Quadril
3.
Microbiology (Reading) ; 164(11): 1457-1470, 2018 11.
Artigo em Inglês | MEDLINE | ID: mdl-29957170

RESUMO

The initial adaptive transcriptional response to nitrogen (N) starvation in Escherichia coli involves large-scale alterations to the transcriptome mediated by the transcriptional activator, NtrC. One of these NtrC-activated genes is yeaG, which encodes a conserved bacterial kinase. Although it is known that YeaG is required for optimal survival under sustained N starvation, the molecular basis by which YeaG benefits N starved E. coli remains elusive. By combining transcriptomics with targeted metabolomics analyses, we demonstrate that the methionine biosynthesis pathway becomes transcriptionally dysregulated in ΔyeaG bacteria experiencing sustained N starvation. It appears the ability of MetJ, the master transcriptional repressor of methionine biosynthesis genes, to effectively repress transcription of genes under its control is compromised in ΔyeaG bacteria under sustained N starvation, resulting in transcriptional derepression of MetJ-regulated genes. Although the aberrant biosynthesis does not appear to be a contributing factor for the compromised viability of ΔyeaG bacteria experiencing sustained N starvation, this study identifies YeaG as a novel regulatory factor in E. coli affecting the transcription of methionine biosynthesis genes under sustained N starvation.


Assuntos
Proteínas de Escherichia coli/genética , Escherichia coli/metabolismo , Regulação Bacteriana da Expressão Gênica/genética , Metionina/biossíntese , Nitrogênio/metabolismo , Proteínas Serina-Treonina Quinases/genética , Transcrição Gênica/genética , Apoproteínas/genética , Escherichia coli/genética , Deleção de Genes , Proteínas PII Reguladoras de Nitrogênio/genética , Proteínas Repressoras/genética , Fatores de Transcrição/genética
4.
Reprod Biomed Online ; 36(2): 145-153, 2018 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-29233502

RESUMO

Previous studies have suggested an association between high intake of sweetened beverages and a number of adverse health outcomes. In this cross-sectional study, we investigated the association between daily consumption of sweetened soft drinks or coffee and the outcome of intracytoplasmic sperm injection (ICSI) treatment. Patients (n = 524) were interviewed by a nutritionist before ICSI treatment, using a food frequency questionnaire. Regression analysis showed that consumption of ≥3 servings of regular soft drinks or any amount of diet soft drinks was associated with oocyte dysmorphism, diminished embryo quality on days 2 and 3 of culture, and a mild effect on blastocyst formation, implantation and pregnancy rate. Consumption of artificially sweetened coffee was negatively associated with embryo quality on days 2 and 3. However, consumption of coffee or soft drinks was not associated with the odds of live birth. Even so, patients should be advised about the potential negative effects of sugar and artificial sweeteners before attempting infertility treatment. This study is limited by the use of a non-validated food frequency questionnaire, lack of information on quantity of sweeteners consumed, and lack of data on glucose levels in blood serum or follicular fluid. Further investigation is warranted.


Assuntos
Bebidas Gaseificadas/efeitos adversos , Café/efeitos adversos , Oócitos/efeitos dos fármacos , Injeções de Esperma Intracitoplásmicas/efeitos dos fármacos , Edulcorantes/efeitos adversos , Adulto , Estudos Transversais , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Estudos Retrospectivos , Injeções de Esperma Intracitoplásmicas/estatística & dados numéricos , Adulto Jovem
5.
Andrologia ; 50(9): e13090, 2018 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-30019480

RESUMO

This prospective-cohort study aimed at investigating the influence of paternal lifestyle factors on semen parameters and intracytoplasmic sperm injection (ICSI) outcomes. The influence of paternal lifestyle factors on seminal quality and ICSI outcomes was investigated in male patients undergoing conventional semen analysis. Cigarette smoking negatively influenced semen volume (B: -0.417, slope: 1.570, p = 0.047), sperm count/ml (B: -7.363, slope: 52.298, p = 0.014), total sperm count (B: -4.43, slope: 178.165, p = 0.023), total motile sperm count (B: -1.38, slope: 100.276, p = 0.045) and SDF (B: 0.014, slope: 9.767, p = 0.033). Alcohol consumption negatively influenced sperm count/ml (B: -12.527, slope: 42.255, p = 0.040) and sperm DNA fragmentation (B: 5.833, slope: 9.680, p = 0.002). There were no significant influences of other paternal lifestyle factors. Cigarette smoking negatively influenced the fertilisation rate (B: -1.349, slope: 21.950, p = 0.039) and the blastocyst formation rate (B: -14.244, slope: 28.851, p = 0.025). Alcohol consumption negatively influenced fertilisation rate (B: -3.617, slope: 20.138, p = 0.041) and blastocyst formation rate (B: -34.801, slope: 30.044, p = 0.042). Cigarette smoking and alcohol consumption appear to reduce semen quality, fertilisation and blastocyst formation rates; thus, it would be wise to recommend that male partners reconsider their lifestyle during in vitro reproduction treatment.


Assuntos
Consumo de Bebidas Alcoólicas/efeitos adversos , Exposição Ambiental/efeitos adversos , Fumar/efeitos adversos , Injeções de Esperma Intracitoplásmicas/estatística & dados numéricos , Espermatozoides , Adulto , Exercício Físico , Feminino , Humanos , Estilo de Vida , Masculino , Gravidez , Estudos Prospectivos , Análise do Sêmen
6.
J Assist Reprod Genet ; 33(12): 1571-1583, 2016 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-27614633

RESUMO

PURPOSE: The goal for the present study was to implement a technique for protein extraction and identification in human cumulus cells (CCs). METHODS: Forty samples of CCs were collected after ovum pick-up from patients undergoing intracytoplasmic sperm injection (ICSI). Samples were split into the blastocyst group (n = 10), including patients in which all embryos converted into blastocysts, and the non-blastocyst group (n = 10), including patients in which none of the embryos reached the blastocyst stage or the positive-pregnancy (n = 10) and negative-pregnancy group (n = 10). Proteins were extracted and injected into a liquid chromatography system coupled to a mass spectrometer. The spectra were processed and used to search a database. RESULTS: There were 87 different proteins in samples from the blastocyst and non-blastocyst groups, in which 30 were exclusively expressed in the blastocyst group and 17 in the non-blastocyst group. Among the 72 proteins detected in the pregnancy groups, 19 were exclusively expressed in the positive, and 16 were exclusively expressed in the negative-pregnancy group. CONCLUSIONS: CC proteomics may be useful for predicting pregnancy success and the identification of patients that should be included in extended embryo culture programs.


Assuntos
Blastocisto/metabolismo , Células do Cúmulo/metabolismo , Desenvolvimento Embrionário/genética , Biossíntese de Proteínas/genética , Blastocisto/fisiologia , Cromatografia Líquida , Células do Cúmulo/fisiologia , Transferência Embrionária/métodos , Feminino , Fertilização in vitro , Regulação da Expressão Gênica no Desenvolvimento , Humanos , Oócitos/metabolismo , Oócitos/fisiologia , Gravidez , Proteômica
7.
Reprod Biomed Online ; 31(1): 30-8, 2015 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-25982093

RESUMO

The aim of this study was to evaluate the influence of patients' lifestyle factors and eating habits on embryo development. A total of 2659 embryos recovered from 269 patients undergoing intracytoplasmic sperm injection cycles were included. The frequency of intake of food items and social habits were registered and its influences on embryo development evaluated. The consumption of cereals, vegetables and fruits positively influenced the embryo quality at the cleavage stage. The quality of the embryo at the cleavage stage was also negatively correlated with the consumption of alcoholic drinks and smoking habits. The consumption of fruits influenced the likelihood of blastocyst formation, which was also positively affected by the consumption of fish. Being on a weight-loss diet and consumption of red meat had a negative influence on the likelihood of blastocyst formation. The likelihood of blastocyst formation was also negatively influenced by the consumption of alcoholic drinks and by smoking habits. The consumption of red meat and body mass index had a negative effect on the implantation rate and the likelihood of pregnancy. In addition, being on a weight-loss diet had a negative influence on implantation rate. Our evidence suggests a possible relationship between environmental factors and ovary biology.


Assuntos
Comportamento , Blastocisto/fisiologia , Dieta , Ingestão de Alimentos , Estilo de Vida , Injeções de Esperma Intracitoplásmicas , Embrião de Mamíferos/citologia , Embrião de Mamíferos/fisiologia , Desenvolvimento Embrionário , Feminino , Humanos , Fenômenos Fisiológicos da Nutrição Materna , Análise de Regressão , Resultado do Tratamento
8.
Zygote ; 23(5): 662-8, 2015 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-25062512

RESUMO

The aim of the present study was to determine the chance of pregnancy and the risk of multiple pregnancies taking into account the number and quality of transferred embryos in patients >36 years old or ≤36 years old. For this study, 1497 patients undergoing intra-cytoplasmic sperm injection (ICSI) cycles in a private assisted reproduction centre were split into groups according to the number and quality of the transferred embryos on the third or fifth day of development. The pregnancy rate and multiple pregnancy rate were compared between the embryo quality groups in patients <36 years old or ≥36 years old. In patients <36 years old, for the day 3 embryo transfer, no significant difference was noted in the pregnancy rate when the groups were compared. However the multiple pregnancy rate was increased by the transfer of an extra low-quality embryo (17.1 versus 28.2%, P = 0.020). For day 5 embryo transfer, the transfer of an extra blastocyst significantly increased the pregnancy rate (36.0 versus 42.4%, P < 0.001) and the multiple pregnancy rate (4.4 versus 16.9%, P < 0.001). In older patients, no significant difference was noted in the pregnancy rate when the groups were compared. However, when an extra low-quality embryo was transferred, a significantly increased rate of multiple pregnancies was observed for day 3 (18.2 versus 26.4%, P = 0.049) and day 5 embryo transfers (5.2 versus 16.1%, P < 0.001). In conclusion, the transfer of an extra low-quality embryo may increase the risk of a multiple pregnancy. In younger patients, the transfer of an extra low-quality blastocyst may also increase the chance of pregnancy.


Assuntos
Transferência Embrionária/métodos , Fertilização in vitro , Taxa de Gravidez , Gravidez Múltipla/estatística & dados numéricos , Injeções de Esperma Intracitoplásmicas , Adulto , Fatores Etários , Blastocisto/citologia , Blastocisto/fisiologia , Feminino , Humanos , Gravidez , Estudos Retrospectivos
9.
J Assist Reprod Genet ; 32(3): 445-50, 2015 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-25595539

RESUMO

PURPOSE: To compare the outcomes of ICSI and IMSI in women presenting with poor ovarian response. METHODS: Data of IMSI cycles performed from January 2011 to December 2013 were included in this retrospective cohort study. Patients were divided into two groups: normoresponder patients (NR group; patients with > 4 oocytes retrieved) and poor-responder patients (PR group; patients with ≤ 4 oocytes retrieved). Patients who underwent IMSI were matched with patients who underwent ICSI in the same period. The ICSI and IMSI outcomes were compared in the NR and PR groups. RESULTS: A total of 414 matched cycles were included in this study. The NR group comprised 324 cycles (164 ICSI and 160 IMSI cycles), and the PR group comprised 90 cycles (43 ICSI and 47 IMSI cycles). In the NR group, no significant differences were observed between the ICSI- and IMSI-treated couples regarding cycle outcomes. In the PR group, fertilisation rate was significantly lower in IMSI-treated couples (53.9% ± 36.7% vs. 79.8% ± 29.3%). The proportion of cycles with embryo transfer (57.4 vs. 79.1%) and the number of transferred embryos (1.5 ± 0.8 vs. 1.9 ± 0.7) were significantly lower in IMSI compared with ICSI. Implantation, pregnancy and miscarriage rates were similar when ICSI or IMSI were performed. CONCLUSIONS: Our results suggest that unselected couples undergoing ICSI that present with poor ovarian response to controlled ovarian stimulation do not benefit from sperm selection under high magnification prior to ICSI.


Assuntos
Fertilização/fisiologia , Injeções de Esperma Intracitoplásmicas , Espermatozoides/fisiologia , Aborto Espontâneo , Adulto , Transferência Embrionária , Feminino , Humanos , Infertilidade Masculina , Masculino , Oócitos/patologia , Oócitos/fisiologia , Gravidez , Taxa de Gravidez
10.
Int Braz J Urol ; 41(4): 757-63, 2015.
Artigo em Inglês | MEDLINE | ID: mdl-26401870

RESUMO

PURPOSE: To investigate whether the semen quality of men undergoing conventional semen analysis is deteriorating over time. MATERIALS AND METHODS: We analyzed and compared the sperm count, motility and morphology of 2300 semen samples provided by males undergoing conventional seminal analysis, from years 2000 to 2002 and 2010 to 2012. The incidences of severe oligozoospermia and azoospermia over time were also compared. RESULTS: A total of 764 sperm samples were analyzed in 2000-2002 and 1536 in 2010-2012. Over time, the mean sperm concentration/ml decreased significantly from 61.7 million in 2000-2002 to 26.7 million in 2010-2012 (R2 = 11.4%, p < 0.001), the total sperm concentration decreased significantly from 183.0 million to 82.8 million (R2 = 11.3%, p < 0.001), and the percentage of normal forms decreased significantly from 4.6% to 2.7% (R2 = 9.8%, p < 0.001). The incidence of severe oligozoospermia significantly increased from 15.7% to 30.3% (OR: 1.09, p < 0.001) and the incidence of azoospermia increased from 4.9% to 8.5% (OR: 1.06, p = 0.001). CONCLUSIONS: This study demonstrated a significant time-related decline in semen quality of infertile patients. This finding might have implications on fertility and emphasizes the need for further studies addressing subject's life-style in order to find and reduce the causative agents. Future prospective and multicenter studies including representative samples of the general population are needed to confirm whether semen quality is really declining.


Assuntos
Infertilidade Masculina/epidemiologia , Análise do Sêmen/estatística & dados numéricos , Contagem de Espermatozoides , Adulto , Azoospermia/epidemiologia , Brasil/epidemiologia , Humanos , Incidência , Masculino , Pessoa de Meia-Idade , Oligospermia/epidemiologia , Análise de Regressão , Estudos Retrospectivos , Motilidade dos Espermatozoides , Fatores de Tempo
11.
PLoS Pathog ; 8(6): e1002743, 2012.
Artigo em Inglês | MEDLINE | ID: mdl-22719249

RESUMO

Cell stress and infection promote the formation of ubiquitinated aggregates in both non-immune and immune cells. These structures are recognised by the autophagy receptor p62/sequestosome 1 and are substrates for selective autophagy. The intracellular growth of Salmonella enterica occurs in a membranous compartment, the Salmonella-containing vacuole (SCV), and is dependent on effectors translocated to the host cytoplasm by the Salmonella pathogenicity island-2 (SPI-2) encoded type III secretion system (T3SS). Here, we show that bacterial replication is accompanied by the formation of ubiquitinated structures in infected cells. Analysis of bacterial strains carrying mutations in genes encoding SPI-2 T3SS effectors revealed that in epithelial cells, formation of these ubiquitinated structures is dependent on SPI-2 T3SS effector translocation, but is counteracted by the SPI-2 T3SS deubiquitinase SseL. In macrophages, both SPI-2 T3SS-dependent aggregates and aggresome-like induced structures (ALIS) are deubiquitinated by SseL. In the absence of SseL activity, ubiquitinated structures are recognized by the autophagy receptor p62, which recruits LC3 and targets them for autophagic degradation. We found that SseL activity lowers autophagic flux and favours intracellular Salmonella replication. Our data therefore show that there is a host selective autophagy response to intracellular Salmonella infection, which is counteracted by the deubiquitinase SseL.


Assuntos
Autofagia , Proteínas de Bactérias/metabolismo , Endopeptidases/metabolismo , Células Epiteliais/parasitologia , Infecções por Salmonella/metabolismo , Salmonella enterica/enzimologia , Animais , Linhagem Celular , Citosol/metabolismo , Citosol/parasitologia , Humanos , Immunoblotting , Imunoprecipitação , Macrófagos/parasitologia , Camundongos , Microscopia Confocal , Microscopia de Fluorescência , Microscopia Imunoeletrônica , Ubiquitina/metabolismo , Vacúolos/metabolismo , Vacúolos/parasitologia
12.
J Assist Reprod Genet ; 31(8): 1053-7, 2014 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-24952886

RESUMO

PURPOSE: To determine whether Brazilian egg donation treatment outcomes with oocytes donated from infertile couples are equivalent to those obtained worldwide with oocytes donated from fertile egg-donors. METHODS: In this descriptive study, egg-donation cycles from 259 women, performed from January 2009 to July 2013, were evaluated. Oocytes were obtained from patients undergoing ICSI who decided to donate their surplus oocytes. We described the survival, fertilization, blastocyst, implantation and pregnancy rates obtained in our infertile donor-recipient program. In addition, we described the results obtained in previous published studies. RESULTS: In our egg-donation program we obtained a fertilization rate of 72.9 %, a blastocyst formation rate of 53.2 %, an implantation rate of 31.1 % and the estimated clinical pregnancy rate per warmed oocyte was 5.4 %. The analyzed studies, performed between 2008 and 2013, included varying numbers of egg-donors (range: 20-600), warmed oocytes (range: 123-3826) and survival rates (range: 85.6-92.5 %). Fertilization rates ranged from 74.2 to 87.0 %, blastocyst formation rate ranged from 41.3 % to 68.0 %, implantation rates ranged from 24.7 % to 55.3 % and the clinical pregnancy rate per warmed oocyte ranged from 3.9 % to 9.8 %. CONCLUSIONS: New and reassuring information derived from our egg-donation program demonstrates outcomes similar to those reported for other egg donation programs.


Assuntos
Criopreservação , Infertilidade Feminina , Doação de Oócitos , Oócitos/citologia , Oócitos/transplante , Bancos de Tecidos , Obtenção de Tecidos e Órgãos , Adulto , Brasil , Implantação do Embrião , Transferência Embrionária , Feminino , Fertilização in vitro , Humanos , Gravidez , Taxa de Gravidez , Adulto Jovem
13.
J Assist Reprod Genet ; 31(8): 1105-10, 2014 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-24893729

RESUMO

PURPOSE: To evaluate: (i) the influence of morphology at cleavage stage on blastocyst formation and implantation, and (ii) whether the transfer of low-quality embryos on day-three would be a better approach than the transfer at blastocyst stage. METHODS: This study included 8,444 embryos obtained from 1,125 patients undergoing ICSI cycles between January/2011 and September/2013. The influence of the quality of the embryo on days-two and -three on blastocyst formation and implantation success was evaluated. Moreover, the implantation potential of low-quality embryos, at cleavage stage, transferred on day-three was compared with the implantation potential of low-quality embryos, at cleavage stage, transferred on day-five. RESULTS: Low-quality embryos on day-two had an approximate 20 % decreased chance of achieving the blastocyst stage, and blastocysts derived from low-quality embryos on day-two had a nearly 40 % decrease in the implantation chance. Low-quality embryos on day-three had a 30 % decreased chance of achieving the blastocyst stage, and blastocysts derived from low-quality embryos on day-three had an almost 40 % decreased implantation chance. The implantation rate didn't differ when low-quality embryos on the cleavage stage were transferred on day-three or left in culture and transferred on day-five. CONCLUSIONS: The transfer of low-quality embryos on day-three is a better approach than transfer at the blastocyst stage. In addition, the embryo morphology evaluation at the cleavage stage is still needed for the selection of the embryo with the best implantation potential in extended embryo culture programmes.


Assuntos
Blastocisto , Fase de Clivagem do Zigoto/transplante , Implantação do Embrião/fisiologia , Transferência Embrionária , Desenvolvimento Embrionário/fisiologia , Fertilização in vitro , Feminino , Humanos , Gravidez , Taxa de Gravidez , Prognóstico
14.
J Assist Reprod Genet ; 31(11): 1533-9, 2014 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-25186503

RESUMO

PURPOSE: To investigate the predictive value of the motile sperm organelle morphology examination (MSOME) on embryo morphology. METHODS: The morphologies of 540 embryos obtained from 60 couples undergoing ICSI were evaluated from days 1 to 5 of development and were examined for associations with the percentages of morphologically normal paternal sperm and of the paternal sperm with large nuclear vacuoles (LNVs) as determined by MSOME. RESULTS: An increased percentage of LNV sperm was associated with increased odds of a zygote presenting with pronuclear abnormalities. It was also associated with decreased odds of (i) normal cleavage on days 2 and 3 of development, (ii) the presence of a high-quality embryo on day 3, (iii) the development of an embryo to the blastocyst stage, and (iv) an embryo possessing a normal trophectoderm and inner cell mass. The calculated areas under the curves differed for the embryos that did and did not develop to the blastocyst stage and for the high- and low-quality blastocysts. The optimal cut-off value for the percentage of LNV sperm that maximised proper blastocyst formation was ≤24.5 %, and the cut-off value that maximised blastocyst quality was ≤19.5 %. CONCLUSIONS: These results suggest a very early onset of paternal influences on embryo development. The evaluation of the incidence of vacuoles by MSOME may significantly improve upon the prognostic information provided by conventional semen analyses.


Assuntos
Desenvolvimento Embrionário , Fertilização , Injeções de Esperma Intracitoplásmicas , Espermatozoides/ultraestrutura , Adulto , Blastocisto/citologia , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Curva ROC , Análise de Regressão , Vacúolos/ultraestrutura
15.
J Assist Reprod Genet ; 31(3): 307-12, 2014 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-24352355

RESUMO

PURPOSE: To investigate if there is a correlation between the prevalence of sperm with large nuclear vacuoles (LNV) and intracytoplasmic sperm injection (ICSI) outcomes. METHODS: Two hundred male patients undergoing ICSI had their sperm morphology evaluated through motile sperm organelle morphology examination (MSOME) and the percentage of LNV sperm was recorded and correlated to the ICSI outcomes. RESULTS: The percentage of sperm with LNV negatively influenced the blastocyst formation (S: 16.9, R(2): 20.5%, p = 0.004) and implantation (S: 34.7, R(2): 26.2%, p = 0.001). There were significant differences in the percentage of sperm with LNV between patients in which pregnancy was achieved or not (22.2% vs. 28.4%, p < 0.001) and in patients with ongoing pregnancy or not (22.4% vs. 28.5%, p < 0.001). The incidence of sperm with LNV was determinant to the decreased odds of pregnancy (OR: 0.74, p < 0.001) and increased odds of miscarriage (OR: 1.46, p < 0.001). The area under the curve (AUC) was sufficient to distinguish between couples which did achieve pregnancy or not (AUC: 0.922, p < 0.001). CONCLUSIONS: The MSOME is a prognostic tool in the prediction of ICSI success and could be used to select patients that should have their sperm selected by MSOME for ICSI.


Assuntos
Análise do Sêmen , Injeções de Esperma Intracitoplásmicas/métodos , Espermatozoides/ultraestrutura , Vacúolos/ultraestrutura , Adulto , Implantação do Embrião , Desenvolvimento Embrionário , Feminino , Humanos , Masculino , Gravidez , Espermatozoides/crescimento & desenvolvimento
16.
Reproduction ; 145(5): 453-62, 2013 May.
Artigo em Inglês | MEDLINE | ID: mdl-23404850

RESUMO

This study has evaluated the performance of a multivariate statistical model to predict embryo implantation potential by processing data from the chemical fingerprinting of culture medium samples used for human embryo culture. The culture medium for 113 embryos from 55 patients undergoing ICSI was collected after embryo transfer. The samples were split into positive (n=29) and negative (n=84) implantation groups according their implantation outcomes (100% or 0% implantation). The samples were individually diluted and analyzed by electrospray ionization mass spectrometry (ESI-MS). The m/z ratios and relative abundances of the major ions in each spectrum were considered for partial least square discriminant analysis. Data were divided into two subsets (calibration and validation), and the models were evaluated and applied to the validation set. A total of 5987 ions were observed in the groups. The multivariate statistical model described more than 82% of the data variability. Samples of the positive group were correctly identified with 100% probability and negative samples with 70%. The culture media used for embryos that were positive or negative for successful implantation showed specific biochemical signatures that could be detected in a fast, simple, and noninvasive way by ESI-MS. To our knowledge, this is the first report that uses MS fingerprinting to predict human embryo implantation potential. This biochemical profile could help the selection of the most viable embryo, improving single-embryo transfer and thus eliminating the risk and undesirable outcomes of multiple pregnancies.


Assuntos
Ectogênese , Implantação do Embrião , Transferência Embrionária , Proteínas Fetais/análise , Modelos Biológicos , Zigoto/metabolismo , Adulto , Calibragem , Estudos de Casos e Controles , Meios de Cultivo Condicionados/química , Análise Discriminante , Feminino , Proteínas Fetais/química , Proteínas Fetais/metabolismo , Humanos , Infertilidade Feminina/etiologia , Infertilidade Feminina/terapia , Infertilidade Masculina/fisiopatologia , Masculino , Pessoa de Meia-Idade , Mapeamento de Peptídeos , Espectrometria de Massas por Ionização por Electrospray , Injeções de Esperma Intracitoplásmicas
17.
J Assist Reprod Genet ; 30(6): 849-54, 2013 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-23547020

RESUMO

PURPOSE: To compare the results of intracytoplasmic morphologically selected sperm injection (IMSI) between cycles in which the swim-up (SUP) or the density gradient centrifugation (DGC) techniques were used for sperm preparation. METHODS: We evaluated 70 IMSI cycles performed in women with age ≤ 37 years, undergoing IMSI as result of male factor. The couples were divided into two groups: DGC group (n = 26) and SUP group (n = 44). The groups were compared with regard to IMSI outcomes. RESULTS: There were no significant differences between SUP and DGC groups regarding the number of follicles, oocytes, mature oocytes, oocyte yield and mature oocyte rate. Fertilization rate and high-quality embryos rate on day 5 of development were similar between SUP and DGC groups. Implantation, pregnancy and miscarriage rates were not statistically different between SUP and DGC groups (28.8 vs 33.3 %, 46.2 vs 57.1 % and 8.3 vs 4.2 %, respectively). CONCLUSIONS: Both the SUP and the DGC techniques recover improved sperm fractions and result in similar IMSI outcomes. Further randomized trials analyzing both the quality of sperm through MSOME and the IMSI outcomes are needed to elucidate the role of sperm preparation techniques and morphology on IMSI outcomes.


Assuntos
Implantação do Embrião , Fertilização in vitro , Injeções de Esperma Intracitoplásmicas/métodos , Espermatozoides/citologia , Aborto Espontâneo , Adulto , Centrifugação com Gradiente de Concentração , Feminino , Humanos , Técnicas de Maturação in Vitro de Oócitos , Infertilidade Masculina/genética , Infertilidade Masculina/patologia , Masculino , Oócitos/citologia , Gravidez , Taxa de Gravidez
18.
J Assist Reprod Genet ; 30(10): 1327-32, 2013 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-23934020

RESUMO

PURPOSE: To (i) investigate a possible association between different features of pronuclear (PN) morphology and different features of blastocyst morphology, (ii) evaluate the combination of PN and blastocyst morphologies as a predictive factor for ICSI outcomes and (iii) identify possible contributing factors to poor PN morphology. METHODS: This study included 908 normally fertilised zygotes reaching full blastocyst stage, obtained from 350 patients undergoing ICSI cycles, in which the implantations rates were 0 % or 100 %. The influence of PN morphology on blastocyst morphology and on the rates of pregnancy and miscarriage was investigated. Embryos were graded and split into three groups, taking into consideration both the PN and the blastocyst status. The pregnancy rate was compared among these groups. RESULTS: Inner cell mass (ICM) alterations were correlated with the number of nucleolar precursor bodies (NPB), while trophectoderm alterations were correlated with the size of the pronuclei and the distribution of the NPB. The distribution of the NPB had an impact on the chances of pregnancy. A significant difference was observed among the groups regarding the pregnancy rate. The maternal age, number of aspirated follicles and number of retrieved oocytes influenced the incidence of PN defects. CONCLUSIONS: These findings suggest that a lower oocyte yield may lead to higher-quality PN zygotes. In addition, different PN features may influence further embryo development, especially the quality of the blastocyst. Moreover, the association between PN and blastocyst morphology may be used as a prognostic tool for implantation.


Assuntos
Implantação do Embrião , Transferência Embrionária , Oócitos/transplante , Aborto Espontâneo , Adulto , Blastocisto , Núcleo Celular/metabolismo , Fase de Clivagem do Zigoto , Feminino , Humanos , Nascido Vivo , Recuperação de Oócitos , Gravidez , Taxa de Gravidez , Prognóstico
19.
Artigo em Inglês | MEDLINE | ID: mdl-37045689

RESUMO

High rates of multiple implantation after assisted reproductive technology (ART) treatment represent one of the major problems for both mothers and their fetuses. Given the availability of techniques intended to identify embryos with the highest chance for development to term, such as comprehensive chromosome screening (CCS) and blastocyst transfer, the decision on the number of embryos to transfer deserves careful consideration. This report presents real-life data from two clinics using the Fischer protocol for cycle programming in patients undergoing ART. Our data indicate that ovarian stimulation using the Fischer protocol provides consistent and optimal ART outcomes in centers following strict quality management standards. However, high multiple implantation rates were observed in fresh and frozen transfer cycles after transferring two embryos - even in patients aged over 39 years. The live birth rates after CCS were superior to those using untested embryos. These findings were held for the three age groups irrespective of the CCS culture day (D1 = PN stages, or D5 = blastocysts). Our results support a single embryo transfer policy, particularly in women under 34 years of age with favorable conditions during ART treatment, i.e., a high number of available fertilized oocytes.


Assuntos
Transferência Embrionária , Técnicas de Reprodução Assistida , Feminino , Humanos , Transferência Embrionária/métodos , Implantação do Embrião/fisiologia , Transferência de Embrião Único , Cromossomos
20.
Nanomaterials (Basel) ; 13(17)2023 Aug 26.
Artigo em Inglês | MEDLINE | ID: mdl-37686937

RESUMO

Organic-inorganic hybrids (OIHs) are a type of material that can be obtained using the sol-gel process and has the advantages of organic and inorganic moieties in a single material. Polyetheramines have been widely used in the preparation of this type of material, particularly in combination with epoxy-based alkoxysilanes. Nevertheless, epoxyciclohexylethyltrimethoxysilane (ECHETMS) is a promising alkoxysilane with an epoxy terminal group that is quite unexplored. In this work, four novel OIH materials were synthesized using the sol-gel method. The OIHs were based on Jeffamines® of different molecular weights (D-230, D-400, ED-600, and ED-900), together with ECHETMS. The materials were characterized using multinuclear solid state NMR, FTIR, BET, UV/Vis spectroscopy, EIS, and TGA. The influence of the Jeffamine molecular weight and the suitability of these materials to act as a supporting matrix for heteroaromatic probes were assessed and discussed. The materials show interesting properties in order to be applied in a wide range of sensing applications.

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