Dereplication Tools for Rhizophora mangle Extracts from Different Mangrove Areas and their Potential Against Staphylococcus aureus.

Rhizophora extracts have several potential biological activities, and their metabolites can be used in the pharmaceutical industry. Extracts of Rhizophora species obtained from mangroves have shown prospective activity against Staphylococcus aureus. This study aimed to investigate the chemical profile of Rhizophora mangle leaves from fringe, basin, and transition mangrove zones and their bactericidal/bacteriostatic potential against S. aureus. R. mangle leaves were collected monthly in 2018 from litterfall in three different zones of the mangrove of Guaratiba State Reserve: fringe, basin, and transition. Extracts were prepared from the material collected in October and December for LC-HRMS/MS analysis, and dereplication was performed using a molecular library search and the classical molecular networking GNPS platform. The minimum inhibitory concentrations (MICs) of the aqueous extract of R. mangle against S. aureus were determined. No S. aureus growth was observed compared to the control for extracts collected from September to December. Different compounds were annotated in each region, yet a marked presence of phenolic compounds was noted, among them glycosylated flavonoid derivatives of quercetin and kaempferol. The results suggest bactericidal/bacteriostatic activity for extracts of R. mangle leaves collected in 2018 from three mangrove forest zones.

Real-world evidence of the burden of sickle cell disease: a 5-year longitudinal study at a Brazilian reference center.

INTRODUCTION: Sickle cell disease (SCD) is an inherited and multisystem blood disorder characterized by hemolytic anemia, vaso-occlusive crises (VOCs), progressive multiorgan damage and increased mortality. In Brazil, it is one of the most common monogenic diseases afflicting 60,000 to 100,000 individuals, however, there are sparse epidemiological data, as well as information on the utilization of public healthcare resources. METHOD: This was a 5-year (2016 - 2020) retrospective study conducted at one Brazilian reference center on SCD - Santa Casa de Sao Paulo, in Sao Paulo, Brazil. RESULTS: Among a total of 100 eligible adult patients, the median age was 31.0 years old, 84% of the patients were aged between 18 and 45 years old; 59% were women and 91% presented the genotype HbSS. The number of hematologist and non-hematologist visits at the outpatient unit were 2,198 and 1,436, respectively. The number of hospital ER visits was 758, of which 51% required 864 days of hospitalization. The main cause for seeking hospital medical care was the VOCs. The numbers and ratios of VOCs were: 1 to 10 VOCs, 64%; 11 to 20, 15%, and; 21 or more, 1%. There was a statistically significant difference between the number of VOCs and hospitalizations, as well as infection. CONCLUSION: Results indicate the burden of SCD on Brazilian patients' daily lives, the impact of VOCs on public healthcare resources, the importance of having a national surveillance program to improve resource utilization and clinical outcomes of patients with SCD and the urgent need for the revitalizing of the current national comprehensive SCD care programs.

Elevated Anti-Müllerian Hormone as a Prognostic Factor for Poor Outcomes of In Vitro Fertilization in Women with Polycystic Ovary Syndrome.

Women with polycystic ovary syndrome (PCOS) tend to have elevated anti-Müllerian hormone (AMH) levels, which appear to correlate with disease severity and pregnancy outcomes. This was a retrospective observational study designed to assess the relationship between circulating AMH levels and in vitro fertilization (IVF) outcomes. The study involved 150 women with PCOS who underwent IVF treatments. The women's IVF cycles were allocated into three subgroups according to AMH levels: 'low' (AMH < 3.7 ng/mL; n = 49), 'middle' (AMH 3.7-7.4 ng/mL; n = 94), and 'high' (AMH > 7.4 ng/mL; n = 56). All pregnancy-related outcomes (positive beta human chorionic gonadotropin (ßHCG), clinical pregnancy rate, live birth rate, and cumulative live birth rate) were greater in women's IVF cycles with 'low' AMH when compared to those with 'middle' or 'high' AMH (p < 0.05). AMH levels below 3.7 ng/mL were found to be associated with lower oocyte immaturity rate and better pregnancy outcomes, although baseline AMH was not shown to have any significant predictive power for live birth and cumulative live birth in the multivariable logistic regression analysis after adjusting for possible confounders nor in the ROC analyses. In summary, the current study lays the groundwork to validate high AMH levels as a poor prognostic factor for pregnancy outcomes after IVF in women with PCOS.

Evaluation of the Antioxidant, Antimicrobial, and Anti-Biofilm Effects of the Stem Bark, Leaf, and Seed Extracts from Hymenaea courbaril and Characterization by UPLC-ESI-QTOF-MS/MS Analysis.

Currently, biofilm-forming bacteria are difficult to treat by conventional antibiotic therapy and are, thus, becoming a clinical and epidemiological problem worldwide. Medicinal plants have been identified as novel alternative treatments due to their therapeutic and antimicrobial effects. In this context, the present study aimed to determine the total phenolic content, antioxidant capacity, and antimicrobial and anti-biofilm potential of nine extracts of Hymenaea courbaril (Fabaceae), popularly known as Jatobá. Furthermore, extracts that exhibited biofilm inhibitory activity against S. aureus (ATCC 25923) were selected for UPLC-HRMS/MS chemical analysis. Our results showed a high total phenolic content, mainly in the stem bark extract, and that the plant is rich in compounds with antioxidant activity. In the anti-biofilm analysis, leaf extracts stood out in comparison with chloramphenicol, with inhibition percentages of 78.29% and 78.85%, respectively. Through chemical analysis by UPLC-HRMS/MS, chrysoeriol-7-O-neohesperidoside, isorhamnetin-3-O-glucoside, and 3,7-di-O-methylquercetin were annotated for the first time in the leaves of H. courbaril. Therefore, these results showed the potential use of H. courbaril as an antioxidant and point to its use in antimicrobial therapy with an anti-biofilm effect.

Physical fitness level and the risk of severe COVID-19: A systematic review.

To verify systematically the association between the status of physical fitness and the risk of severe Coronavirus disease 2019 (COVID-19). This systematic review is in accordance with the Preferred Reporting Items for Systematic Review and Meta Analyses (PRISMA) statement and the eligibility criteria followed the Population, Intervention, Comparison, Outcomes and Study (PICOS) recommendation. PubMed, Embase, SciELO and Cochrane electronic databases were searched. All studies that explored the relationship between the pattern of physical fitness and COVID-19 adverse outcomes (hospitalization, intensive care unit admission, intubation, or mortality), were selected. The quality of the studies was assessed by the specific scale of the Newcastle-Ottawa Scale. A total of seven observational studies were identified in this systematic review; 13 â€‹468 patients were included in one case-control study, two cohort studies, and four cross-sectional studies. All studies reported an inverse association between high physical fitness and severe COVID-19 (hospitalization, intensive care admission, or mortality). Only some studies reported comorbidities, especially obesity and cardiovascular disorders, but the results remained unchanged after controlling for comorbidities. The quality of the seven studies included was moderate according to the Newcastle-Ottawa Quality Assessment Scale. The methodological heterogeneity of the studies included did not allow a meta-analysis of the findings. In conclusion, higher physical fitness levels were associated with lower risk of hospitalization, intensive care admissions, and mortality rates among patients with COVID-19.

Antiparasitic properties of 4-nerolidylcatechol from Pothomorphe umbellata (L.) Miq. (Piperaceae) in vitro and in mice models with either prepatent or patent Schistosoma mansoni infections.

ETHNOPHARMACOLOGICAL RELEVANCE: Roots of Pothomorphe umbellata (L.) Miq. are used in traditional medicine of Africa and South America for the treatment of malaria and helminthiasis. However, neither P. umbellata nor its isolated compounds have been evaluated against Schistosoma species. AIMS OF THIS STUDY: To investigate the antischistosomal effects of P. umbellata root extracts and the isolated compound 4-nerolidylcatechol (4-NC) against Schistosoma mansoni ex vivo and in murine models of schistosomiasis. MATERIALS AND METHODS: The crude hydroalcoholic (PuE) and hexane (PuH) extracts of P. umbellata roots were prepared and initially submitted to an ex vivo phenotypic screening against adult S. mansoni. PuH was analyzed by HPLC-DAD, characterized by UHPLC-HRMS/MS, and submitted to chromatographic fractionation, leading to the isolation of 4-NC. The anthelmintic properties of 4-NC were assayed ex vivo against adult schistosomes and in murine models of schistosomiasis for both patent and prepatent S. mansoni infections. Praziquantel (PZQ) was used as a reference compound. RESULTS: PuE (EC50: 18.7 µg/mL) and PuH (EC50: 9.2 µg/mL) kill adult schistosomes ex vivo. The UHPLC-HRMS/MS analysis of PuH, the most active extract, revealed the presence of 4-NC, peltatol A, and peltatol B or C. After isolation from PuH, 4-NC presented remarkable in vitro schistosomicidal activity with EC50 of 2.9 µM (0.91 µg/mL) and a selectivity index higher than 68 against Vero mammalian cells, without affecting viability of nematode Caenorhabditis elegans. In patent S. mansoni infection, the oral treatment with 4-NC decreased worm burden and egg production in 52.1% and 52.3%, respectively, also reducing splenomegaly and hepatomegaly. 4-NC, unlike PZQ, showed in vivo efficacy against juvenile S. mansoni, decreasing worm burden in 52.4%. CONCLUSIONS: This study demonstrates that P. umbellata roots possess antischistosomal activity, giving support for the medicinal use of this plant against parasites. 4-NC was identified from P. umbellata roots as one of the effective in vitro and in vivo antischistosomal compound and as a potential lead for the development of novel anthelmintics.

All that glitters is not gold: a stereological study of human donor oocytes.

Here we report a quantitative analysis of human metaphase II (MII) oocytes from a 22-year-old oocyte donor, retrieved after ovarian-controlled hyperstimulation. Five surplus donor oocytes were processed for transmission electron microscopy (TEM), and a stereological analysis was used to quantify the distribution of organelles, using the point-counting technique with an adequate stereological grid. Comparisons between means of the relative volumes (Vv) occupied by organelles in the three oocyte regions, cortex (C), subcortex (SC) and inner cytoplasm (IC), followed the Kruskal-Wallis test and Mann-Whitney U-test with Bonferroni correction. Life cell imaging and TEM analysis confirmed donor oocyte nuclear maturity. Results showed that the most abundant organelles were smooth endoplasmic reticulum (SER) elements (26.8%) and mitochondria (5.49%). Significant differences between oocyte regions were found for lysosomes (P = 0.003), cortical vesicles (P = 0.002) and large SER vesicles (P = 0.009). These results were quantitatively compared with previous results using prophase I (GV) and metaphase I (MI) immature oocytes. In donor MII oocytes there was a normal presence of cortical vesicles, SER tubules, SER small, medium and large vesicles, lysosomes and mitochondria. However, donor MII oocytes displayed signs of cytoplasmic immaturity, namely the presence of dictyosomes, present in GV oocytes and rare in MI oocytes, of SER very large vesicles, characteristic of GV oocytes, and the rarity of SER tubular aggregates. Results therefore indicate that the criterion of nuclear maturity used for donor oocyte selection does not always correspond to cytoplasmic maturity, which can partially explain implantation failures with the use of donor oocytes.

In silico screening of phenylethanoid glycosides, a class of pharmacologically active compounds as natural inhibitors of SARS-CoV-2 proteases.

Since the advent of Covid-19, several natural products have been investigated regarding their in silico interactions with SARS-CoV-2 proteases - 3CLpro and PLpro, two of the most important pharmacological targets for antiviral development. Phenylethanoid glycosides (PG) are a class of natural products present in important medicinal plants and a drug containing this group of active ingredients has been successfully used in the treatment of Covid-19 in China. Thus, a dataset with 567 derivatives of this class was built from reviews published between 1994 and 2020, and their interaction against both SARS-CoV-2 proteases was investigated. The virtual screening was performed by filtering the PGs through the evaluation of scores based on the AutoDock Vina, GOLD/ChemPLP, and GOLD/GoldScore evaluation functions. The bRO5 pharmacokinetic parameters of the PGs ranked in the previous step were analyzed and their interaction with key amino acid residues of the 3CLpro and PLpro enzymes was evaluated. Ninety-eight compounds were identified by computational approaches against PLpro and 80 PGs against 3CLpro. Of these, four interacted with key catalytic residues of PLpro, which is an indicative of inhibitory activity, and three compounds interacted with catalytic key residues of 3CLpro. Of these, five PGs occur in plants of the Traditional Chinese Medicine (TCM), while two are components of plants/formulations currently used in the Covid-19 protocols in China. The data presented here show the potential of PGs as selective inhibitors of SARS-CoV-2 3CLpro and PLpro.

Disentangling the effects of terroir, season, and vintage on the grapevine fungal pathobiome.

The composition, diversity and dynamics of microbial communities associated with grapevines may be influenced by various environmental factors, including terroir, vintage, and season. Among these factors, terroir stands out as a unique possible determinant of the pathobiome, the community of plant-associated pathogens. This study employed high-throughput molecular techniques, including metabarcoding and network analysis, to investigate the compositional dynamics of grapevine fungal pathobiome across three microhabitats (soil, woody tissue, and bark) using the Furmint cultivar. Samples were collected during late winter and late summer in 2020 and 2021, across three distinct terroirs in Hungary's Tokaj wine region. Of the 123 plant pathogenic genera found, Diplodia, Phaeomoniella, and Fusarium displayed the highest richness in bark, wood, and soil, respectively. Both richness and abundance exhibited significant disparities across microhabitats, with plant pathogenic fungi known to cause grapevine trunk diseases (GTDs) demonstrating highest richness and abundance in wood and bark samples, and non-GTD pathogens prevailed soil. Abundance and richness, however, followed distinct patterns Terroir accounted for a substantial portion of the variance in fungal community composition, ranging from 14.46 to 24.67%. Season and vintage also contributed to the variation, explaining 1.84 to 2.98% and 3.67 to 6.39% of the variance, respectively. Notably, significant compositional differences in fungi between healthy and diseased grapevines were only identified in wood and bark samples. Cooccurrence networks analysis, using both unweighted and weighted metrics, revealed intricate relationships among pathogenic fungal genera. This involved mostly positive associations, potentially suggesting synergism, and a few negative relationships, potentially suggesting antagonistic interactions. In essence, the observed differences among terroirs may stem from environmental filtering due to varied edaphic and mesoclimatic conditions. Temporal weather and vine management practices could explain seasonal and vintage fungal dynamics. This study provides insights into the compositional dynamics of grapevine fungal pathobiome across different microhabitats, terroirs, seasons, and health statuses. The findings emphasize the importance of considering network-based approaches in studying microbial communities and have implications for developing improved viticultural plant health strategies.

Abiotic environmental factors drive the diversity, compositional dynamics and habitat preference of ectomycorrhizal fungi in Pannonian forest types.

Ectomycorrhizal (ECM) fungi are among the most diverse and dominant fungal groups in temperate forests and are crucial for ecosystem functioning of forests and their resilience toward disturbance. We carried out DNA metabarcoding of ECM fungi from soil samples taken at 62 sites in the Bükk Mountains in northern Hungary. The selected sampling sites represent the characteristic Pannonian forest types distributed along elevation (i.e., temperature), pH and slope aspect gradients. We compared richness and community composition of ECM fungi among forest types and explored relationships among environmental variables and ECM fungal alpha and beta diversity. The DNA sequence data generated in this study indicated strong correlations between fungal community composition and environmental variables, particularly with pH and soil moisture, with many ECM fungi showing preference for specific zonal, topographic or edaphic forest types. Several ECM fungal genera showed significant differences in richness among forest types and exhibited strong compositional differences mostly driven by differences in environmental factors. Despite the relatively high proportions of compositional variance explained by the tested environmental variables, a large proportion of the compositional variance remained unexplained, indicating that both niche (environmental filtering) and neutral (stochastic) processes shape ECM fungal community composition at landscape level. Our work provides unprecedented insights into the diversity, landscape-level distribution, and habitat preferences of ECM fungi in the Pannonian forests of Northern Hungary.

Exploring Relationships among Grapevine Chemical and Physiological Parameters and Leaf and Berry Mycobiome Composition.

Improving our knowledge on biotic and abiotic factors that influence the composition of the grapevine mycobiome is of great agricultural significance, due to potential effects on plant health, productivity, and wine characteristics. Here, we assessed the influence of scion cultivar on the diversity and composition of fungal communities in the berries and leaves of three different cultivars. We generated DNA metabarcoding data, and statistically compared the richness, relative abundance, and composition of several functional groups of fungi among cultivars, which are partly explained by measured differences in chemical composition of leaves and berries and physiological traits of leaves. Fungal communities in leaves and berries show contrasting patterns among cultivars. The richness and relative abundance of fungal functional groups statistically differ among berry and leaf samples, but less so among cultivars. Community composition of the dominant functional groups of fungi, i.e., plant pathogens in leaves and saprotrophs in berries, differs significantly among cultivars. We also detect cultivar-level differences in the macro- and microelement content of the leaves, and in acidity and sugar concentration of berries. Our findings suggest that there appears to be a relatively diverse set of fungi that make up the grapevine mycobiome at the sampled terroir that spans several cultivars, and that both berry and leaf mycobiomes are likely influenced by the chemical characteristics of berries and leaves, e.g., pH and the availability of nutrients and simple carbohydrates. Finally, the correlation between fungal community composition and physiological variables in leaves is noteworthy, and merits further research to explore causality. Our findings offer novel insights into the microbial dynamics of grapevine considering plant chemistry and physiology, with implications for viticulture.

Bioassay-Guided Fractionation of Siparuna glycycarpa n-Butanol Extract with Inhibitory Activity against Influenza A(H1N1)pdm09 Virus by Centrifugal Partition Chromatography (CPC).

Siparuna glycycarpa occurs in the Amazon region, and some species of this genus are used in Brazilian folk medicine. A recent study showed the inhibitory effect of this species against influenza A(H1N1)pdm09 virus, and in order to acquire active fractions, a polar solvent system n-butanol-methanol-water (9:1:10, v/v) was selected and used for bioassay-guided fractionation of n-butanol extract by centrifugal partition chromatography (CPC). The upper phase was used as stationary phase and the lower phase as mobile (descending mode). Among the collected fractions, the ones coded SGA, SGC, SGD, and SGO showed the highest antiviral inhibition levels (above 74%) at 100 µg·mL-1 after 24 h of infection. The bioactive fractions chemical profiles were investigated by LC-HRMS/MS data in positive and negative ionization modes exploring the Global Natural Products Social Molecular Networking (GNPS) platform to build a molecular network. Benzylisoquinoline alkaloids were annotated in the fractions coded SGA, SGC, and SGD collected during elution step. Aporphine alkaloids, O-glycosylated flavonoids, and dihydrochalcones in SGO were acquired with the change of mobile phase from lower aqueous to upper organic. Benzylisoquinolinic and aporphine alkaloids as well as glycosylated flavonoids were annotated in the most bioactive fractions suggesting this group of compounds as responsible for antiviral activity.

Influence of Women´s Residence Region on Assisted Reproduction Treatments - Experience of a Tertiary Center in Northern Portugal.

OBJECTIVE: Data on the possible influence of women´s region of residence, within the same country, on the outcomes of medically assisted reproduction cycles are scarce. This study aims to assess the impact of the women's region of residence on the results of in-vitro fertilization cycles. METHODS: We evaluated in-vitro fertilization cycles between 2010 and 2017, performed in a northern Portugal assisted reproduction center. We defined two groups: Douro Litoral (group 1; n=783), and Trás-os-Montes and Alto Douro (group 2; n=178). We analyzed demographics and cycle-related variables, and we calculated the rates for embryo transfer cycles. We used the Mann-Whitney and Chi-square tests and p<0.05 was considered statistically significant. RESULTS: We included 961 cycles. The region of residence had no effect on the following variables: women´s age; body mass index; or duration of infertility (p>0.05). Group 2 had a statistical significant lower number of previous cycles than group 1 (1.3±0.5 and 1.5±0.7; p=0.005). In the sub-analysis of cycles with embryo transfer (n=781), group 1 obtained had rates of normal fertilization (62.5% vs. 57.5%; p=0.04), miscarriage rate (30.0 vs. 10.9%; p=0.007), and lower implantation rates compared to group 2 (33.3% vs. 50.0%; p<0.001). CONCLUSIONS: Women from the region of Trás-os-Montes e Alto Douro had a lower number of previous cycles, compared to those from the Douro Litoral, despite the absence of statistical significant differences in terms of age or infertility duration. These findings reinforce the need to contemplate the sociodemographic and socioeconomic variables in this context.

Flavonoids from Siparuna cristata as Potential Inhibitors of SARS-CoV-2 Replication.

The novel coronavirus SARS-CoV-2 has been affecting the world, causing severe pneumonia and acute respiratory syndrome, leading people to death. Therefore, the search for anti-SARS-CoV-2 compounds is pivotal for public health. Natural products may present sources of bioactive compounds; among them, flavonoids are known in literature for their antiviral activity. Siparuna species are used in Brazilian folk medicine for the treatment of colds and flu. This work describes the isolation of 3,3',4'-tri-O-methyl-quercetin, 3,7,3',4'-tetra-O-methyl-quercetin (retusin), and 3,7-di-O-methyl-kaempferol (kumatakenin) from the dichloromethane extract of leaves of Siparuna cristata (Poepp. & Endl.) A.DC., Siparunaceae, using high-speed countercurrent chromatography in addition to the investigation of their inhibitory effect against SARS-CoV-2 viral replication. Retusin and kumatakenin inhibited SARS-CoV-2 replication in Vero E6 and Calu-3 cells, with a selective index greater than lopinavir/ritonavir and chloroquine, used as control. Flavonoids and their derivatives may stand for target compounds to be tested in future clinical trials to enrich the drug arsenal against coronavirus infections. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s43450-021-00162-5.

Semantic Adaptation and Validation of the Stanford Hypnotic Susceptibility Scale, Form C, in the Chilean Population.

One of the difficulties of evaluating hypnotizability in Chile is the limited existence of validated instruments. In this study, the Mexican version of Stanford Hypnotic Susceptibility Scale, Form C, was semantically adapted and validated. A descriptive cross-sectional study was carried out in 102 Chilean university students. The content validation was performed by 3 experts; the internal consistency was evaluated with KR-20. The difficulty of the items was measured with a difficulty index. The majority of the students were classified with high hypnotizability. The mean score obtained was 7.41 (SD = 1.84). The internal consistency was acceptable (KR-20 = 0.73). The item with the least difficulty was arm lowering, whereas the auditory hallucination was the item with the greatest difficulty. The survey showed metric properties to be considered as a valid and reliable instrument to measure the level of hypnotizability in the Chilean population.

Amazonian Siparuna extracts as potential anti-influenza agents: Metabolic fingerprinting.

ETHNOPHARMACOLOGICAL RELEVANCE: Siparuna species are used in Brazilian Folk Medicine for the treatment and prophylaxis of colds, fever, headache, gastrointestinal disorders and rheumatic pain. AIM OF THE STUDY: This study aimed to investigate a possible anti-influenza activity of 25 extracts from leaves of Amazonian S. cristata, S. decipiens, S. glycycarpa, S. reginae and S. sarmentosa based on their folk medicinal uses as well as to investigate their metabolic fingerprinting. The chemical composition of the active extracts was further dereplicated. MATERIAL AND METHODS: The chemical composition of the crude EtOH extracts from five Siparuna species were investigated by ESI (±) LC-QTOF-MS2. Organic extracts were obtained by liquid-liquid partition with solvents of increasing polarity, generating 25 extracts which were subjected to a quick DI-ESI (±) IT-MS fingerprint analysis. These extracts were tested against influenza virus replication and cellular toxicity using MDCK cells and influenza A/Michigan/45/2015 (H1N1)pdm09 virus. The compounds in the active BuOH extracts from S. glycycarpa and S. sarmentosa were annotated by ESI (±) LC-QTOF-MS2. RESULTS: Analysis of the EtOH extracts revealed the presence of alkaloids and flavonoids, in the positive and negative ionization modes. Out of the 25 organic extracts screened for their antiviral activity, the BuOH extracts from S. glycycarpa and S. sarmentosa were the most active, inhibiting 96.0 ± 1.3% and 89.5 ± 0.8% of influenza virus replication 24 h post-infection. These inhibitory effects were maintained until 72hpi. Alkaloids, O- and C-flavonoid glycosides, dihydrochalcones and a procyanidin dimer were annotated in these extracts. CONCLUSIONS: The inhibitory effect against influenza A(H1N1)pdm09 virus replication shown by Amazonian Siparuna species corroborates the use of these plants in Brazilian Folk Medicine, showing their potential as anti-influenza agents. These promising results stimulate the continuation of this study with the aim of isolating the compound(s) responsible for this bioactivity, thus contributing to a better knowledge of those species and to the research of natural products with potential anti-influenza activity.

Zika virus seroprevalence in women who gave birth during Zika virus outbreak in Brazil - a prospective observational study.

BACKGROUND: The recent Zika virus (ZIKV) outbreak in Brazil started in August 2015 and ended in May 2017 without effective public health measures for its control have been taken. The immunological status of a community may not only predict future outbreaks as well to answer questions regarding ZIKV not known yet. OBJECTIVE: To verify the seroprevalence of ZIKV in a group of women who were pregnant during the Zika virus outbreak in Recife, three to nine months after the delivery, and to evaluate the neurodevelopment of their children. METHODS: A cross-sectional study enrolled participants of a cohort study held at Instituto de Medicina Integral Professor Fernando Figueira (IMIP) during the ZIKV outbreak in Recife. Mothers who gave birth between the last trimester of 2015 and the first semester of 2016, period of the peak of microcephaly outbreak in Recife, were invited. All participants had the serum tested by the anti-ZIKV IgG/IgM enzyme-liked immunosorbent assays, ELISA kit (Euroimmun, Lübeck, Germany). All children whose mothers presented positive serology for ZIKV performed the IgG/IgM ELISA test for ZIKV. These children were also evaluated by a neuropediatrician and the Denver II development screening test was applied. RESULTS: Among the 132 studied pregnant women who gave birth at the peak of ZIKV outbreak in Recife, all were ZIKV IgM negative and 81 (61,3%) had ZIKV IgG positive. Mothers ZIKV IgG positive had more fever and rash during the pregnancy as compared with mothers negative for ZIKV; respectively 27/81 (33,3%) vs 6/51 (11,7%), p = 0.005 and 22/81 (27,2%) vs 4 (7,8%), p = 0.016. Only one child had IgG positive serology for ZIKV. No children had neurodevelopment defect for the age group and the Denver II normal scores. CONCLUSIONS: A high ZIKV IgG seroprevalence in pregnant women at the end of the ZIKV outbreak in Recife was found. This finding suggests that community protective immunity may have contributed to the end of ZIKV outbreak in Recife, Brazil.

Stereological study of organelle distribution in human oocytes at metaphase I.

We have previously presented a stereological analysis of organelle distribution in human prophase I oocytes. In the present study, using a similar stereological approach, we quantified the distribution of organelles in human metaphase I (MI) oocytes also retrieved after ovarian stimulation. Five MI oocytes were processed for transmission electron microscopy and a classical manual stereological technique based on point-counting with an adequate stereological grid was used. Kruskal-Wallis and Mann-Whitney U-tests with Bonferroni correction were used to compare the means of relative volumes (Vv) occupied by organelles. In all oocyte regions, the most abundant organelles were mitochondria and smooth endoplasmic reticulum (SER) elements. No significant differences were observed in Vv of mitochondria, dictyosomes, lysosomes, or SER small and medium vesicles, tubular aggregates and tubules. Significant differences were observed in other organelle distributions: cortical vesicles presented a higher Vv (P = 0.004) in the cortex than in the subcortex (0.96% vs 0.1%) or inner cytoplasm (0.96% vs 0.1%), vesicles with dense granular contents had a higher Vv (P = 0.005) in the cortex than in the subcortex (0.1% vs 0%), and SER large vesicles exhibited a higher Vv (P = 0.011) in the inner cytoplasm than in the subcortex (0.2% vs 0%). Future stereological analysis of metaphase II oocytes and a combined quantitative data of mature and immature oocytes, will enable a better understanding of oocyte organelle distribution during in vivo maturation. Combined with molecular approaches, this may help improve stimulation protocols and in vitro maturation methods.

Recovery of bioactive compounds from white grape (Vitis vinifera L.) stems as potential antimicrobial agents for human health.

The grape is a matrix rich in bioactive compounds and its production generates large quantities of by-products, such as grape stems, which, to date, present low commercial value. However, there is a growing interest in the application of this material as a source of phenolic compounds. Therefore, the present study aims at assessing the phytochemical profile of (poly)phenolic extracts of white Portuguese grape stem varieties produced in the Região Demarcada do Douro (Portugal). The antioxidant activity determined by several assays, as well as the antimicrobial activity using the disc diffusion method against human gastrointestinal pathogenic bacteria of the hydromethanolic extracts, were evaluated. This work presents very positive results as the rich composition in phenolic compounds (94.71-123.09 mg GA-1 and 0.02-73.79 mg g-1 for the total phenol content and for individual phenolics, respectively) presented by grape stems can explain the high antioxidant (0.37-1.17 mmol Trolox g-1) and antimicrobial activities against, essentially, Gram-positive bacteria, and in some cases with higher efficacy than commercial antibiotics. Thus, demonstrating that this wine by-product should deserve greater attention from the pharmaceutical industries due to its excellent biological properties and characteristics not yet applied.