The Ala307Thr polymorphism of the follicle-stimulating hormone receptor (FSHR) gene is associated with the dose of recombinant FSH received during IVF/ICSI treatment.

OBJECTIVE: Follicle-stimulating hormone (FSH) is essential for folliculogenesis, acting through the follicle-stimulating hormone receptor (FSHR) that is present on the membrane of granulosa cells. Polymorphisms in the FSHR gene may lead to an altered pattern of receptor expression on the cell surface or to changes in affinity for FSH. The aim of this prospective study was to detect any association between the follicle-stimulating hormone receptor (FSHR) gene Ala307Thr polymorphism (rs6165) and ovarian reserve, ovarian response or clinical results in IVF/ICSI treatment. METHODS: This prospective cohort study included 450 women who underwent IVF/ICSI cycles. DNA was extracted from peripheral blood, and the Ala307Thr FSHR polymorphism (rs6165) was genotyped using the TaqMan SNP genotyping assay. Participants were divided into three groups according to their Ala307Thr FSHR genotype: Thr/Thr (n:141), Thr/Ala (n=213) and Ala/Ala (n=96). The results were tested for associations with age, anti-Mullerian hormone (AMH) levels, antral follicle count (AFC), total dose of r-FSH, follicle size, number of retrieved oocytes, and clinical outcome of IVF/ICSI cycles. The statistical analyses were performed using Fisher's exact test and the Kruskal‒Wallis test. RESULTS: An association between the genotype of the FSHR (Ala307Thr) polymorphism and the dose of r-FSH was observed. Patients with the Ala/Ala genotype received a higher r-FSH dose than patients with the Ala/Thr (p=0.0002) and Thr/Thr (p=0.02) genotypes. No other correlation was observed. CONCLUSION: The Ala/Ala genotype was associated with the use of higher doses of recombinant FSH (r-FSH), suggesting that homozygosis of this allelic variant (Ala) provides lower sensitivity to r-FSH.

Covid-19 school closures negatively impacted elementary-school students' reading comprehension and reading fluency skills.

In Brazil, schools remained closed to prevent the spread of COVID-19 for approximately two years. In this study, we investigated how COVID-19 school closures were associated with gaps and losses in Brazilian students' reading skills. We contrasted the reading comprehension and fluency of 2nd- to 4th-graders in 2022 with a 2019 cohort. Furthermore, we investigated how 2022 5th graders' reading comprehension and fluency skills have improved since 2019. Overall, results showed that COVID-19 school closures were associated with gaps but not losses in reading comprehension and fluency. Therefore, these skills should be targets of educational interventions.

Milonine attenuates the lipopolysaccharide-induced acute lung injury in mice by modulating the Akt/NF-κB signaling pathways.

Acute lung injury is an inflammation that triggers acute respiratory distress syndrome with perialveolar neutrophil infiltration, alveolar-capillary barrier damage, and lung edema. Activation of the toll-like receptor 4 complex (TLR4/MD2) and its downstream signaling pathways are responsible for the cytokine storm and cause alveolar damage. Due to the complexity of this pulmonary inflammation, a defined pharmacotherapy has not been established. Thus, this study evaluated the anti-inflammatory potential of milonine, an alkaloid of Cissampelos sympodialis Eichl, in an experimental model of lung inflammation. BALB/c mice were lipopolysaccharide-challenged and treated with milonine at 2.0 mg/kg. Twenty-four hours later, the bronchoalveolar fluid, peripheral blood, and lungs were collected for cellular and molecular analysis. The milonine treatment decreased the cell migration (mainly neutrophils) to the alveoli, the pulmonary edema, and the cytokine levels (IL-1ß, IL-6, TNF-α). The systemic IL-6 level was also reduced. The milonine docking analysis demonstrated hydrophobic interaction at TLR4/MD2 groove with Ile124 and Phe126 amino acids. Indeed, the alkaloid downregulated the kinase-Akt and NF-κB through TLR4/MD2. Therefore, milonine is an effective inflammatory modulator being a potential molecule for the treatment of lung inflammation.

Impact of Intracytoplasmic Morphologically Selected Sperm Injection (IMSI) on Birth Defects: A Systematic Review and Meta-Analysis.

OBJECTIVE: It is generally accepted that the incidence of birth defects in spontaneously conceived children ranges between 2.0-4.0%. However, several studies have shown that babies born after assisted reproductive technology (ART) procedures tend to present more congenital malformations than naturally conceived children, with 6.5% of the children born after intracytoplasmic sperm injection (ICSI) presenting birth defects. The use of high magnification sperm selection before ICSI was introduced in the early 2000s to allow the identification of spermatozoa with low risk of sperm DNA damage. Intracytoplasmic morphologically selected sperm injection (IMSI) is expected to change the incidence of congenital malformations, although data on the incidence of birth defects in children conceived after IMSI are still scarce. METHODS: A systematic review based on searches performed in electronic databases (PubMed, EMBASE, Web of Science, SCOPUS, and Cochrane Central Register of Controlled Trials) including articles published by February 2021 was conducted to identify trials comparing the neonatal outcomes of ICSI and IMSI. The outcome measured was the rate of birth defects in children born after ICSI or IMSI. Three trials were included as targets for data extraction and meta-analysis. RESULTS: Our meta-analysis included 3907 children conceived after IMSI (1280) or ICSI (2627). The incidence of birth defects was statistically different, with 2.5% (32/1280) in IMSI and 4.5% (119/2627) in ICSI (RR=0.59; 95% CI=0.40-0.87; p=0.007). The results demonstrated that IMSI decreased the incidence of structural defects compared to ICSI - 2.2% (18/830) vs. 3.8% (78/2049) - in a statistically significant manner (RR=0.58; 95%CI=0.35-0.96; p=0.04). No significant difference was observed in chromosomal abnormalities (Trisomy 13; 18; 21 and Triple X) between children conceived after IMSI (8/830) or ICSI (19/2049) (RR=1.07; 95%CI=0.47-2.43; p=0.87). CONCLUSIONS: IMSI seems to be an effective tool at reducing the incidence of structural defects compared to ICSI. However, IMSI does not change the incidence of chromosomal abnormalities.

Os processos educativos em grupos comunitários como caminhos para a promoção da saúde e da segurança alimentar e nutricional na Atenção Básica / Educational processes in community groups as pathways to Health Promotion and Food and Nutrition Security in Primary Care

O presente trabalho buscou explicitar saberes e práticas da Promoção da Saúde (PS) e da Segurança Alimentar e Nutricional (SAN) no âmbito da Atenção Básica (AB), partindo de processos educativos em grupos comunitários pautados pela perspectiva da Educação Popular (EP). O processo investigativo baseou-se na sistematização de experiências, tendo como foco atividades desenvolvidas no período de dez anos (2007-2017) pelo Programa de Extensão "Práticas Integrais de Promoção da Saúde e Nutrição na Atenção Básica (PINAB)", o qual é vinculado à Universidade Federal da Paraíba e é construído de forma compartilhada com trabalhadores e usuários de uma Unidade de Saúde da Família. Empregou-se a pesquisa documental como forma de mobilização de fontes para essa sistematização. Como resultado, pôde- se apontar: os pressupostos que sustentaram essa experiência, as formas de organização da mesma, os desafios e as potencialidades de seus processos educativos.

The present work sought to explain knowledge and practices of Health Promotion (PS) and Food and Nutritional Security (SAN) within the scope of Primary Care (AB), starting from educational processes in community groups based on the perspective of Popular Education (EP). The investigative process was based on the systematization of experiences, focusing on activities developed in the ten-year period (2007-2017) by the Extension Program "Comprehensive Practices for the Promotion of Health and Nutrition in Primary Care (PINAB)", which is linked to the Federal University of Paraíba and is built in a shared way with workers and users of a Family Health Unit. Documentary research was used as a way of mobilizing sources for this systematization. As a result, it was possible to point out: the assumptions that sustained this experience, the organization forms of it, the challenges and the potential of its educational processes.

Noninvasive preimplantation genetic testing for aneuploidies (niPGT-A) and the principle of primum non nocere.

Perhaps with the intention of obtaining larger amounts of free-DNA, some groups are routinely postponing and establishing free-DNA collection in culture medium for Noninvasive preimplantation genetic testing for aneuploidies (niPGT-A) to day 6 for all blastocysts. A meta-analysis served as the basis for such decision, since statistically similar live birth rates were observed when the transfers of euploid blastocysts were performed on day 5 versus day 6 However, the euploidy analysis was conducted in only two studies However, after including the two more studies we performed a new meta-analysis that clearly showed the risks of losing live births with the decision of adopting the 6th day as the endpoint for gathering free-DNA. We would be losing 1.71x more live births.

Noninvasive Preimplantation Genetic Testing for Aneuploidy (niPGT-A): The first Brazilian baby.

Recently, a new technology known as the Noninvasive Preimplantation Genetic Testing for Aneuploidy (niPGT-A) emerged, using cell-free DNA present in the spent culture media of human blastocysts. Unlike PGT-A, in which only trophectoderm cells are used, niPGT-A reflects the ploidy state of these cells and internal cell mass, suggesting that this new technology may be less prone to error, being more reliable than the invasive test. The aim of the present study was to report the first occurrence of childbirth following niPGT-A in Brazil.

In Vitro and In Vivo Antimicrobial Activity of Cymbopogon citratus (DC.) Stapf. Against Staphylococcus spp. Isolated from Newborn Babies in an Intensive Care Unit.

Phytotherapy is an emerging topic of health research, with particular focus on studying the efficiency of essential oils as antimicrobials. In this study, we investigated the in vitro and in vivo susceptibility of Staphylococcus strains isolated from newborns to Cymbopogon citratus oil. The in vitro susceptibility of the microorganisms to C. citratus essential oil was compared with the activities of standard antibiotics administered to newborns using disk diffusion and microdilution methods. For in vivo assessment, 30 Wistar rats were wounded and subjected to infection of Staphylococcus aureus strain DRJ080, followed by treatment with the antibiotic vancomycin, C. citratus, or carbopol polymer gel (control) for 11 days. S. aureus accounted for 23.36% of the 107 Staphylococcus sp. strains isolated. Both vancomycin and the essential oil of C. citratus inhibited the growth of all microorganisms in vitro. The minimum inhibitory and bactericidal concentrations for the C. citratus oil were 0.625 mg/mL in all strains tested. The oil had the same therapeutic effectiveness as vancomycin against S. aureus DRJ080 in rats. Thus, Staphylococcus strains of newborns are sensitive to C. citratus oil, both in vitro and in vivo, demonstrating its potential as an antibiotic alternative.

New therapeutic protocol for improvement of endometrial receptivity (PRIMER) for patients with recurrent implantation failure (RIF) - A pilot study.

OBJECTIVE: To evaluate whether or not one should use a new Protocol for Endometrial Receptivity Improvement (PRIMER) based on platelet-rich plasma (PRP) and granulocyte colony-stimulation factor (G-CSF) to enhance ongoing pregnancy rates in patients with recurrent implantation failure (RIF). METHODS: Women undergoing IVF/ICSI were prospectively divided into two groups: - PRIMER/RIF group (n:33): patients with RIF (defined as ≥2embryo transfers (ETs) and at least 5 morphologically good embryos transferred) in which intrauterine PRP injection and subcutaneous G-CSF-injection were performed. - Control group (n:33): patients in their first IVF/ICSI attempt/cycle (without PRP or G-CSF injection). The PRP was prepared using autologous fresh-whole blood processed to increase platelet-concentration in 2 to 4 fold. All patients undergoing the PRP-treatment received 0.7ml of it through intrauterine-injection 48 hours before the ET. G-CSF (300mg/0.5ml) started simultaneously to PRP and was administered subcutaneously every week. RESULTS: Regarding implantation, clinical pregnancy and miscarriage rates, we found no statistically significant difference (18.2% versus 17.6%, p=0.90; 36.4% versus 30.3%, p=0.61 and 25.0% versus 9.0%, p=0.43, respectively). The use of PRIMER enabled RIF patients (previous ET µ: 4.0±1.5) to reach similar ongoing pregnancy and live birth rates like those patients who had their first IVF/ICSI cycle attempt (27.3% versus 27.3%, p=0.99). CONCLUSIONS: Our results showed, for the first time, evidence that this therapeutic protocol (PRIMER) could be used as a feasible treatment based on biological rationale for patients with RIF, considering its promising outcomes, it is a simple procedure and not associated with patient complications.

Randomized comparison of two commercial culture media (Cook and Vitrolife) for embryo culture after IMSI.

OBJECTIVE: A variety of studies randomizing women/cycles or oocytes/embryos has been carried out to compare different culture media for culturing embryos up to cleavage or blastocyst stages showing controversial results. A recent systematic review suggested that data in the literature are insufficient to conclude the best culture medium for embryo quality, pregnancy and implantation. The objective of this study was to evaluate whether there is any difference between two commercial culture media regarding clinical outcomes after IMSI cycles. METHODS: A total of 120 patients, ≤39 years of age, undergoing ART treatment submitted to the IMSI program were prospectively broken down and randomized into two groups: Group I (Cook media) and Group II (Vitrolife media). RESULTS: Our data demonstrated that there was no difference using all the media from Cook or all the media from Vitrolife, for culturing embryos till day 2, in the bench incubator at low O2 concentration, in relation to fertilization, embryo quality, pregnancy and implantation rates (p>0.05). CONCLUSION: Both culture media used, Cook medium and Vitrolife medium, for the IMSI procedure and for later embryo culture with transfer on the second day, are equally effective and can be used depending on the ease and availability of acquisition.

The effects of male age on sperm DNA damage: an evaluation of 2,178 semen samples.

OBJECTIVE: This study aimed to evaluate the effects of male age on sperm DNA damage. METHODS: This cross-sectional study included semen samples collected from 2,178 men seen at an infertility clinic. For DNA integrity analysis, the proportions of spermatozoa showing DNA fragmentation (TUNEL assay), abnormal chromatin packaging/underprotamination (chromomycin A3), abnormal mitochondrial membrane potential (MMP/MitoTracker Green), and apoptosis (annexin V) were recorded. For group comparisons, enrolled subjects were divided into three groups based on their ages: ≤35 years; 36-44 years; and ≥45 years. The associations between age and sperm parameters were assessed using Spearman's rank correlation coefficient. RESULTS: Although aging did not affect sperm apoptosis (p>.05), sperm DNA fragmentation and MMP deteriorated significantly with age (p<.05). Chromatin packaging/protamination improved significantly with age (p<.05). CONCLUSION: Sperm DNA fragmentation worsened with age and was apparently associated with mitochondrial damage. The age-related increase in sperm DNA damage suggests that delaying childbearing, not only in women but also in men, might jeopardize a couple's reproductive capacity. The increase seen in chromatin packaging might represent a protective feature for DNA. However, additional studies must be performed to confirm the results concerning chromatin packaging/protamination.

Semen parameters in men with varicocele: DNA fragmentation, chromatin packaging, mitochondrial membrane potential, and apoptosis.

OBJECTIVE: To evaluate varicocele effects on conventional semen parameters: sperm DNA fragmentation, chromatin packaging, mitochondrial membrane potential (MMP) and sperm apoptosis. METHODS: A cross-sectional study was carried out with semen samples from 2,399 men of couples who attended an infertility clinic. A total of 16.3% (391/2399) of the men were diagnosed with varicocele by a urologist. RESULTS: A regression analysis revealed that the percentages of sperm with DNA fragmentation, abnormal chromatin packaging, and abnormal MMP were significantly increased in individuals with varicocele, when compared to men without varicocele. Apoptosis was not influenced by varicocele. Conventional semen parameters were significantly worse in individuals with the disease. On the other hand, in men with varicocele, Spearman's correlation demonstrated that early apoptosis and abnormal MMP showed a positive and significant correlation with sperm DNA fragmentation. CONCLUSION: Men with varicocele had worse semen parameters, including increased levels of sperm DNA fragmentation, inactive mitochondria, and abnormal chromatin packaging. These changes are possible causes of infertility in individuals with varicocele.

Influence of the abstinence period on human sperm quality: analysis of 2,458 semen samples.

OBJECTIVE: The aim of this study was to evaluate the influence of different periods of abstinence on conventional semen parameters as well as functional parameters in human semen, including mitochondrial function, chromatin packing and sperm DNA fragmentation. METHODS: We recruited a cohort of 2,458 men undergoing infertility investigation. Semen analyses were performed according to WHO guidelines/morphology-motile sperm organelle morphology examination/MSOME. For DNA integrity analysis, the percentages of DNA fragmentation (TUNEL), abnormal chromatin packaging/underprotamination (chromomycin A3/CMA3), abnormal mitochondrial membrane potential (MMP/MitoTracker Green), and apoptosis (annexin-V) were recorded. Associations between the sexual abstinence period and sperm parameters were assessed using Spearman correlation. For group comparisons, the subjects were categorized according to the sexual abstinence period (SAP) into three groups: SAP <2 days, SAP 2-5 days, and SAP >5 days. RESULTS: The duration of abstinence had a statistically significant positive influence on sperm concentration and volume, the number of leukocytes and a statistically significant negative influence on sperm motility and vitality. The percentages of DNA fragmentation and MMP (mitochondrial damage) worsened with the increased duration of abstinence. The percentage of sperm protamination was statistically significantly increased with abstinence. CONCLUSION: Increase in the sexual abstinence period influences sperm quality. This study reinforces the importance of the duration of ejaculatory abstinence on semen parameter variation. It highlights the deleterious effect of increased abstinence on DNA damage, which is most likely associated with ROS (mitochondrial damage/number of leukocytes). The increase in chromatin packaging can represent a protective feature for DNA.

Single versus sequential culture medium: which is better at improving ongoing pregnancy rates? A systematic review and meta-analysis.

This study aimed to evaluate if single medium is better than sequential medium at improving ongoing pregnancy rates in patients undergoing assisted reproductive technology (ART) procedures. The data featured in this meta-analysis were extracted from four randomized controlled trials yielded from a systematic search carried out on electronic databases. The primary endpoint was ongoing pregnancy rate. Secondary endpoints included clinical pregnancy and miscarriage rates. The endpoints for ongoing pregnancy rate were also analyzed based on the time at which the embryo transfers were performed: cleavage stage (day 2/3) and/or blastocyst stage (day 5/6). There were no significant differences between single and sequential medium for clinical pregnancy (RR=1.09; 95%CI=0.83-1.44; p=0.53), ongoing pregnancy (RR=1.11; 95%CI=0.87-1.40; p=0.39), or miscarriage rates (RR=0.89; 95%CI=0.44-1.81; p=0.74). No significant difference was found for ongoing pregnancy rate (RR=1.29; 95%CI=0.93-1.78; p=0.12) between single and sequential medium when only trials in which embryos were transferred at the blastocyst stage were included. In conclusion, the choice of embryo culture approach - single or sequential medium - did not affect the ongoing pregnancy rates of patients undergoing ART cycles.

Fresh embryos versus freeze-all embryos - transfer strategies: Nuances of a meta-analysis.

OBJECTIVE: The present meta-analysis aimed to evaluate whether the freeze-all strategy (Freeze/All-ET) could bring about improvements in the clinical assisted reproductive technique (ART) outcomes when compared with the fresh embryo transfer strategy (Fresh-ET) in patients undergoing an ART cycle in accordance with the mean number of oocytes collected. METHODS: A systematic review based on electronic searches in databases (PubMed, EMBASE, Web of Science, SCOPUS, and Cochrane Central Register of Controlled Trials) was carried out to identify randomized controlled trails (RCTs) comparing ART outcomes between fresh-embryo transfers versus elective frozen-embryo transfers up to February of 2017. Four reviewers independently evaluated abstracts, validity assessment and data extraction. Odds Ratio (OR) values with a 95% confidence interval (CI), and heterogeneity were evaluated. RESULTS: Five RCTs were included as targets for data extraction and meta-analysis purposes. The results of this meta-analysis were divided into two parts (Freeze/All-ET versus Fresh-ET): Part I- All trials in which the mean number of collected oocytes was >12 and <21 for ongoing pregnancy rate (OR=1.24; 95%CI=1.06-1.44), clinical pregnancy rate (OR=1.19; 95%CI=0.98-1.43), live birth rate (OR= 1.39; 95%CI=0.99-1.95), and miscarriage rate (OR=0.68; 95%CI=0.46-1.00); Part II- Three studies where the mean number of oocytes retrieved was >12 and <15 for ongoing pregnancy rate (OR=1.17; 95%CI=1.00-1.38), clinical pregnancy rate (OR=1.34; 95%CI=0.79-2.28), live birth rate (OR= 1.24; 95%CI=1.00-1.55), and miscarriage rate (RR=0.68; 95%CI=0.46-1.02). CONCLUSIONS: The freeze-all strategy could be favorable when high numbers of oocytes are collected, signaling an association between higher ovarian stimulation and consequent impairment of endometrial receptivity. However, when the mean number of oocytes collected is <15, the freeze-all strategy does not appear to be advantageous.

Key performance indicators score (KPIs-score) based on clinical and laboratorial parameters can establish benchmarks for internal quality control in an ART program.

OBJECTIVE: KPIs have been employed for internal quality control (IQC) in ART. However, clinical KPIs (C-KPIs) such as age, AMH and number of oocytes collected are never added to laboratory KPIs (L-KPIs), such as fertilization rate and morphological quality of the embryos for analysis, even though the final endpoint is the evaluation of clinical pregnancy rates. This paper analyzed if a KPIs-score strategy with clinical and laboratorial parameters could be used to establish benchmarks for IQC in ART cycles. METHODS: In this prospective cohort study, 280 patients (36.4±4.3years) underwent ART. The total KPIs-score was obtained by the analysis of age, AMH (AMH Gen II ELISA/pre-mixing modified, Beckman Coulter Inc.), number of metaphase-II oocytes, fertilization rates and morphological quality of the embryonic lot. RESULTS: The total KPIs-score (C-KPIs+L-KPIs) was correlated with the presence or absence of clinical pregnancy. The relationship between the C-KPIs and L-KPIs scores was analyzed to establish quality standards, to increase the performance of clinical and laboratorial processes in ART. The logistic regression model (LRM), with respect to pregnancy and total KPIs-score (280 patients/102 clinical pregnancies), yielded an odds ratio of 1.24 (95%CI = 1.16-1.32). There was also a significant difference (p<0.0001) with respect to the total KPIs-score mean value between the group of patients with clinical pregnancies (total KPIs-score=20.4±3.7) and the group without clinical pregnancies (total KPIs-score=15.9±5). Clinical pregnancy probabilities (CPP) can be obtained using the LRM (prediction key) with the total KPIs-score as a predictor variable. The mean C-KPIs and L-KPIs scores obtained in the pregnancy group were 11.9±2.9 and 8.5±1.7, respectively. Routinely, in all cases where the C-KPIs score was ≥9, after the procedure, the L-KPIs score obtained was ≤6, a revision of the laboratory procedure was performed to assess quality standards. CONCLUSION: This total KPIs-score could set up benchmarks for clinical pregnancy. Moreover, IQC can use C-KPIs and L-KPIs scores to detect problems in the clinical-laboratorial interface.

Social oocyte cryopreservation: a portrayal of Brazilian women.

OBJECTIVE: This study aimed to determine what Brazilian childless women of reproductive age think about oocyte cryopreservation to postpone pregnancy and their reasons for performing or not performing this procedure. METHODS: Women of reproductive age were randomly selected from the general population using different e-mail lists and were invited to participate in the study by completing an online web survey regarding social oocyte cryopreservation. The survey was also distributed through social media to women of reproductive age. RESULTS: Although most of the responders had a partner (86.9%) and had already planned the pregnancy of their first child (69.6%), 85.4% (379) considered the potential of social oocyte freezing to improve their chances of giving birth later in life. Those that had already planned pregnancy were two times more likely to intend to freeze their oocytes (p=0.03). The most important barrier for not undergoing oocyte cryopreservation was cost. The women who indicated that they could not currently undergo the procedure now because of cost were two times (p=0.03) more likely to intend to cryopreserve their oocytes than women who thought that they would not need to delay pregnancy. CONCLUSION: Brazilian women who think that they are not ready to have a family are discovering the option of oocyte cryopreservation. Most participants considered safeguarding their reproductive potential. Making the procedure more accessible could give women the opportunity to make proactive decisions about the future of their fertility.