T. Bonetti | Universidade Federal de São Paulo (UNIFESP) (original) (raw)

Papers by T. Bonetti

Reproductive Sciences, 2014

Objective: To evaluate the expression of endometrial matrix metalloproteinases (MMPs) 2 and 9 and... more Objective: To evaluate the expression of endometrial matrix metalloproteinases (MMPs) 2 and 9 and E-cadherin in periimplantation phase of infertile women who have undergone in vitro fertilization (IVF) cycles. Methods: This prospective study included 51 patients who underwent endometrial biopsy during the receptive phase in a menstrual cycle prior to IVF treatment. The samples were evaluated by tissue microarray for immunohistochemical study. Results: The expression of MMP-2, MMP-9, and E-cadherin in the endometrium prior to IVF treatment was not associated with pregnancy. There was a decrease in E-cadherin immunodetection, the higher the age of the patients, a negative relationship between E-cadherin and MMP-2, and a positive association between MMP-9 and E-cadherin. Conclusions: The MMP-2, MMP-9, and E-cadherin are expressed in the endometrium of infertile patients during the receptive phase of the natural menstrual cycle. However, there is no correlation between the expression of these molecules and the clinical IVF outcomes.

Human Reproduction

Study question Is the morula compaction pattern associated with early zygote cleavage anomalies a... more Study question Is the morula compaction pattern associated with early zygote cleavage anomalies and euploidy rates? Can it represent a possible mechanism of embryonic correction? Summary answer Abnormalities in the first divisions give rise to dysfunctional blastomeres, which were excluded from compaction process. This process represent a possible embryonic correction mechanism. What is known already IVF embryos have historically been analyzed based on morphological characteristics focusing on days 2/3 (cleavage stage) and 5/6 (blastocyst). The morula stage, usually reached on day 4, represent a transitional phase resulting from the embryonic genome activation and has received little attention in clinical embryology. However, during this crucial stage, cell differentiation begins, metabolism reaches higher levels, changes in cell shape and cell-destination mechanisms are enhanced, providing opportunities for “quality control”. At morula stage, embryos with severe genetic abnormaliti...

JBRA Assisted Reproduction

Objective: This work aimed to evaluate the guar gum (GG) effect in cryoprotectant medium on human... more Objective: This work aimed to evaluate the guar gum (GG) effect in cryoprotectant medium on human cryopreserved spermatozoa. Methods: This study was done in an assisted human reproduction clinic, in association with the Tiradentes University and the Institute of Technology and Research (Aracaju/SE-Brazil). All patients who met the inclusion criteria, voluntarily, signed a consent form and were invited to participate in the research, by clarifying the terms and agreeing with the procedures to be carried out. We selected 30 normal seminal samples, collected and analyzed from July to October 2015. The samples were divided into the following groups: (I): TYB ® (Test Yolk Buffer); (II) GEYC (Glycerol Egg-Yolk Citrate); (III) GEYC containing 0,07% GG and (IV) GEYC containing 0,1% GG. After a minimum of 48 hours the samples were thawed and analysed, with fluorescent probes, the mitochondrial activity and the integrity of the plasma and acrosomal membranes of the sperm cells after the cryopreservation process. ANOVA and Tukey test were used and the of the data was performed using the statistical software GraphPad Prisma 5.0. The values were expressed as mean and standard error. Values of P<0.05 were considered statistically significant. Results: The use of GG in groups III (85.67±2.69%) and IV (89.27±1.96%), showed plasma membrane integrity results significantly higher (P<0.05) than the control groups I (54.97±6.79%) and II (52.33±7.06%). The same was observed (P<0.05) regarding the integrity of the acrosomal membrane when groups III (87.12±2.45%) and IV (90.58±1.52%) were compared with the control groups I (54.97±6.79%) and II (52.33±7.08%). In relation to mitochondrial activity, the use of GG in groups III (10.50±1.77%) and IV (8.83±1.71%) showed similar results (P>0.05) to the control groups I (12.27±1.66%) and II (10.83±1.65%). Conclusion: The addition of GG to GEYC improved the results of plasma membrane integrity and acrossome integrity in all cryopreserved samples. These results suggest that GG has a better cryoprotective potential when compared to TYB ® and GEYC used alone. In this way, this gum presents itself as a promising cryoprotectant agent, however, further studies are still needed to confirm the use of this polysaccharide in the cryopreservation oh human spermatozoa.

Brazilian Journal of …, 2007

The introduction of routine vaccination against tetanus and diphtheria in Brazil has decreased th... more The introduction of routine vaccination against tetanus and diphtheria in Brazil has decreased the incidence and changed the epidemiology of both diseases. We then investigated the prevalence of Corynebacterium diphtheriae carrier status and diphtheria and tetanus immunity in São Paulo, Brazil. From November 2001 to March 2003, 374 individuals were tested for the presence of C. diphtheriae in the nasooropharynx and of serum diphtheria and tetanus antibodies. Participants were all healthy individuals without acute or chronic pathologies and they were stratified by age as follows: 0-12 months and 1-4, 5-9, 10-14, 15-24, 25-39, 40-59, and ≥60 years. Antibodies were assessed using a double-antigen ELISA. C. diphtheriae species were identified by biochemical analysis and toxigenicity was assessed by the Elek test. For diphtheria, full protection (antibodies ≥0.1 IU/mL) was present in 84% of the individuals, 15% had basic protection (antibodies ≥0.01 and <0.1 IU/mL) and 1% were susceptible (antibodies <0.01 IU/mL). Full tetanus protection (antibodies ≥0.1 IU/mL) was present in 79% of the participants, 18% had basic protection (antibodies ≥0.01 and <0.1 IU/mL) and 3% were susceptible (antibodies <0.01 IU/mL). The geometric mean of diphtheria and tetanus antibodies reached the highest values at 5-9 years and decreased until the 40-59-year age range, increasing again in individuals over 60 years. Three participants (0.8%) were carriers of C. diphtheriae, all non-toxigenic strains. The present results demonstrate the clear need of periodic booster for tetanus and diphtheria vaccine in adolescents and adults after primary immunization in childhood.

Reproductive BioMedicine Online

Journal of assisted reproduction and genetics, Jan 12, 2018

The risk of ovarian failure after radiotherapy and/or chemotherapy is a concern among oncologic w... more The risk of ovarian failure after radiotherapy and/or chemotherapy is a concern among oncologic women. There is no doubt regarding the psycho-emotional benefits of fertility preservation (FP) after a cancer diagnosis because concerns about biological conception are a source of anxiety and can even affect the patient's cancer recovery. The aim of this study was to evaluate oncology patients' feelings, concerns, and life quality impacts related to FP. This qualitative cross-sectional study was based on a questionnaire administered to a selected group of women diagnosed with cancer who underwent FP. Thirty-four eligible women (23-39 years old) completed this questionnaire. Two of the participants already had a child, and most of them (61.8%) stated a desire to have children at the time of FP. Their feelings primarily involved safety (44.1%) and hope (23.5%). Time and/or financial issues (82.4%) were the main challenge for FP. All of the women noted the importance of FP, with ma...

Journal of Assisted Reproduction and Genetics

Advances in reproductive techniques, mainly the introduction of oocyte vitrification, have provid... more Advances in reproductive techniques, mainly the introduction of oocyte vitrification, have provided the opportunity to conceive from oocyte banks. The aim of this study was to compare the clinical outcomes of fresh and vitrified oocytes in an egg donation program following blastocyst transfer. This retrospective observational study included 504 oocyte donation cycles. All donor women were younger than 30 years of age. The recipient cycles were divided into two groups: fresh oocytes (n = 78) or vitrified oocytes (n = 426). All oocytes were fertilized by ICSI using ejaculated sperm, followed by blastocyst transfer. Endometrium preparation was performed with estradiol valerate plus micronized progesterone according to standard protocols. Recipients were of similar age (fresh 42.0 ± 4.5 years vs vitrified 41.8 ± 4.8 years; p = 0.790). The fresh group received more mature oocytes for injection compared to the vitrified group (10.1 ± 2.8 vs 9.2 ± 2.2; p = 0.005). The two pronuclei (2PN) rate (74.5 vs 77.4%; p = 0.195) and blastocyst rate (48.8 vs 51.6%; 0.329) were similar between the fresh and vitrified groups, respectively. The rates of clinical pregnancy were 60.9% in the fresh and 59.0% in the vitrified groups (p = 0.771). Our findings suggest that vitrified oocytes result in similar pregnancy rates when compared to fresh oocytes with blastocyst transfer in an egg donation program. Moreover, vitrified oocytes may allow for a better cycle schedule, starting with a lower number of oocytes to be fertilized. Therefore, we hypothesize that egg banks with vitrified oocytes could be safely utilized in an egg donation program.

Reproductive BioMedicine Online, 2016

The aneuploidy rates in expanded blastocysts biopsied on days 5 and 6 development were assessed i... more The aneuploidy rates in expanded blastocysts biopsied on days 5 and 6 development were assessed in women undergoing IVF followed by array comparative genomic hybridization. This study included 1171 expanded blastocysts from 465 patients. Among the 465 patients, 215 and 141 underwent embryo biopsy on day 5 and day 6 (46.2% and 30.3%, respectively), and 109 underwent biopsy on both days 5 and 6 (23.4%). The cycles of 206 women were cancelled because only aneuploidy embryos were present (44.3%). The aneuploid embryos were classified according to the type as single, double or complex aneuploidy. No differences were observed in the distributions of these three categories according to the day of the biopsy. The aneuploidy rate was also evaluated according to maternal age, and was found to be higher in older patients; however, no differences in this rate were detected between embryos biopsied on days 5 and 6 according to maternal age. Biopsy was carried out when blastocysts reached the expanded stage. The embryos biopsied on day 6 had a higher rate of aneuploidy (69.9%) than those biopsied on day 5 (61.4%); however, the euploid embryos transferred had similar chances for successful and healthy gestation.

Reproductive sciences (Thousand Oaks, Calif.), Sep 22, 2016

Endometriosis affects approximately 12% of reproductive-age women and is currently diagnosed usin... more Endometriosis affects approximately 12% of reproductive-age women and is currently diagnosed using invasive laparoscopic surgery. Differences in gene expression in the eutopic endometrium between women with and without endometriosis have been reported, and determining the reproducibility of these genetic differences in the endocervical epithelium would represent an important step toward developing novel diagnostic strategies. In this study, we analyzed gene expression in the endocervical epithelium in women with and without moderate or severe endometriosis. Using RT2 Profiler PCR Arrays, we analyzed gene expression in endocervical epithelial cells from women with deep endometriosis (n = 4) and healthy women (n =6). Nine genes were identified as being upregulated: 5 cell cycle genes (cyclin B1 [CCNB1], cyclin G1 [CCNG1], cullin 1 [CUL1], general transcription factor IIH, polypeptide 1 [GTF2H1], and proliferating cell nuclear antigen [PCNA]), 3 cytokine genes (C3, chemokine (C-C motif...

Fertility and Sterility, 2015

J Urol, 2008

by electroporation. Our results suggest that this approach could have great relevance for the stu... more by electroporation. Our results suggest that this approach could have great relevance for the study of sperm function and male infertility.

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Pregnancy hypertension, 2015

Adipose tissue is responsible for triggering systemic inflammatory response and these changes may... more Adipose tissue is responsible for triggering systemic inflammatory response and these changes may be involved in the pathophysiology of preeclampsia. To characterize the lipid profile in the placenta of patients with early-onset preeclampsia. Samples were collected from placenta of 10 pregnant women with early-onset preeclampsia and 10 controls. Lipids were extracted using the Bligh-Dyer protocol and were analyzed by MALDI TOF-TOF mass spectrometry. The placental analysis of patients with preeclampsia revealed the main class glycerophosphoserines-GP03 (PS) as the most prevalent lipid, representing 56.28% of the total composition. Other main class found was macrolides/lactone polyketides-PK04 with 32.77%, both were increased in preeclamptic placentas when compared to the control group, PS (p<0.0001) and PK04 (p<0.0001). Some lipids found in placentas from patients with preeclampsia were reduced when compared to control group; glycerophosphoethanolamines-GP02 (Pet) (p<0.0001)...

Apoptosis : an international journal on programmed cell death, 2015

We hypothesized that p27(kip1) overexpression can regulate endometriosis cell proliferation, apop... more We hypothesized that p27(kip1) overexpression can regulate endometriosis cell proliferation, apoptosis and vascular endothelial growth factor (VEGF) expression in the endometrium. The overexpression of p27(kip1) was obtained by transduction of p27(kip1) in primary cultures of endometrium obtained from women with endometriosis tissue with gene therapy technology. First generation bicistronic adenovirus: AdCMVhp27IRESEGFP (Adp27) and AdCMVNull (AdNull) were engineered in order to induce p27(kip1) expression in endometrial cells primary culture. The effect of p27(kip1) overexpression was elucidated through the cell proliferation evaluation and the expression of the cell cycle-related proteins p16, p21, p27, and p53. Cell cycle and apoptosis in endometrial cells from women with and without endometriosis were also evaluated. The VEGF levels were evaluated 1 and 7 days after transduction. The experiments were performed using Immunofluorescence stainings and flow cytometry technique. The c...

Brazilian Journal of Medical and Biological Research, 2015

Preimplantation genetic diagnosis (PGD) was originally developed to diagnose embryo-related genet... more Preimplantation genetic diagnosis (PGD) was originally developed to diagnose embryo-related genetic abnormalities for couples who present a high risk of a specific inherited disorder. Because this technology involves embryo selection, the medical, bioethical, and legal implications of the technique have been debated, particularly when it is used to select features that are not related to serious diseases. Although several initiatives have attempted to achieve regulatory harmonization, the diversity of healthcare services available and the presence of cultural differences have hampered attempts to achieve this goal. Thus, in different countries, the provision of PGD and regulatory frameworks reflect the perceptions of scientific groups, legislators, and society regarding this technology. In Brazil, several texts have been analyzed by the National Congress to regulate the use of assisted reproduction technologies. Legislative debates, however, are not conclusive, and limited information has been published on how PGD is specifically regulated. The country requires the development of new regulatory standards to ensure adequate access to this technology and to guarantee its safe practice. This study examined official documents published on PGD regulation in Brazil and demonstrated how little direct oversight of PGD currently exists. It provides relevant information to encourage reflection on a particular regulation model in a Brazilian context, and should serve as part of the basis to enable further reform of the clinical practice of PGD in the country.

International journal of fertility & sterility, 2013

This study compares the developmental capacity of gametes retrieved from the largest follicle wit... more This study compares the developmental capacity of gametes retrieved from the largest follicle with small follicles of a cohort in controlled ovarian stimulated cycles. This prospective study performed in a private assisted fertilization center included 1016 follicles collected from 96 patients who underwent intra cytoplasmic sperm injection (ICSI). After follicular aspiration, oocytes were assigned to two groups according to the diameter of the derived follicle. The large follicle group (n=96) comprised oocytes derived from the leading follicle of the cohort and the small follicle group (n=920) consisted oocytes derived from the smaller follicles of the cohort. The fertilization and percentage of topquality embryos were compared between groups by Chi-square or Fisher's exact test, where appropriate. The effect of the follicular diameter on oocyte dimorphism was assessed by binary logistic regression. A significantly higher percentage of oocytes derived from the leading follicle ...