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This study aimed to analyze the effects of physical exercise (PE) on bone health in menopausal women through an umbrella review and to assess the quality of systematic reviews (SRs) and meta-analyses (MAs) included. The review was registered in PROSPERO (CRD42020208130) and the Rayyan application was used. The methodological quality of the included studies was evaluated by A MeaSurement Tool to Assess Systematic Reviews (AMSTAR), and Grading of Recommendations Assessment, Development and Evaluation (GRADE) was applied to assess the level of evidence of the results. The results showed that low-intensity jumping exercises with longer sessions proved to be efficient in the hip segment. Swimming requires further investigation, as it showed high heterogeneity. Aerobic and resistance exercises showed inconsistent results, requiring further studies with these modalities of PE. Concurrent training showed improvements in the lumbar spine, femoral neck, Ward's triangle and trochanter. Finally, combined resistance exercises are effective in preserving bone mineral density (BMD) of the femoral neck and lumbar spine in postmenopausal women. In conclusion, jumping exercises were efficient in the hip, while aerobic and resistance exercises are still inconsistent. Concurrent training showed improvements in BMD of the lumbar spine, femoral neck, Ward's triangle and trochanter. Finally, combined resistance protocols are effective in preserving BMD of the femoral neck and lumbar spine in postmenopausal women.

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Natural products have been used in the treatment of illnesses throughout the history of humankind. Exploitation of bioactive compounds from natural sources can aid in the discovery of new drugs, provide the scaffold of new medicines. In the face of challenging diseases, such as the COVID-19 pandemic, for which there was no effective treatment, nature could offer insights as to novel therapeutic options for control measures. However, the environmental impact and supply chain of bioactive production must be carefully evaluated to ensure the detrimental effects will not outweigh the potential benefits gained. History has already proven that highly bioactive compounds can be rare and not suitable for medicinal exploitation; therefore, the sustainability must be accessed before expensive, time-demanding, and large trials can be initialized. A sustainable option to readily produce a phytotherapy with minimal environmental stress is the use of agro-industry wastes, a by-product produced in high quantities. In this review we evaluate the sustainability issues associated with the production of phytotherapy as a readily available tool for pandemic control.

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AIMS: The objective of this study was to evaluate the cytotoxic activity of Cronobacter strains isolated from foods (n = 50) and clinical samples (n = 6) in Brazil and genotype selected strains (n = 18) using multi-locus sequence typing (MLST) METHODS AND RESULTS: The cytotoxic activity of C. sakazakii (n = 29), C. dublinensis (n = 13), C. malonaticus (n = 6), C. turicensis (n = 6) and C. muytjensii (n = 2) was screened using Vero, RK13, Hep2c, NCTC clone 929 and BHK-21 cell lines. Selected Cronobacter strains were assigned to C. sakazakii ST 21, C. turicensis ST 252, C. sakazakii ST 647, and three newly assigned STs: C. turicensis STs 738-740. The maximum death caused by non-heat-treated filtrates was 20·4, 86·2, 47·0 and 84·0%, in Vero, RK13, Hep2c and NCTC clone 929 cells, respectively. These were caused by C. sakazakii strains C291 and C292 (ST 494) which had been isolated during neonatal Cronobacter meningitis infection, and C110 (ST 395) isolated from flaxseed flour. Thermal treatment (100°C/20 min) significantly reduced the cytotoxicity activity in NCTC clone 929 and Vero cells (P ≤ 2 × 10-6 ), but not in RK13 (P = 0·12) and Hep2c (P = 0·85), indicating the cytotoxin(s) were probably proteinaceous. Electron microscopy revealed that cytotoxic compounds from C. sakazakii induced several cell death characteristics, including loss of cell-cell contact, microvilli reduction and cellular lysis. Autophagic vacuoles and mitochondrial damage were the most common ultrastructural features observed. CONCLUSIONS: It was concluded that Cronobacter strains, especially C. sakazakii, could produce heat-labile cytotoxic compounds in cell filtrates. SIGNIFICANCE AND IMPACT OF THE STUDY: This study providing insights into the pathogenesis of the Cronobacter genus. Cytotoxins were identified in excreted filtrates of C. sakazakii strains isolated from food and clinical specimens. The presence of Cronobacter strains that can produce cytotoxins in foods can be a potential threat to human health and highlight the need for high levels of hygiene.

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The population of Pará (a state in Brazil) has a very characteristic food culture, as a majority of the carbohydrates consumed are obtained from cassava (Manihot esculenta Crantz) derivatives. Tucupi is the boiled juice of cassava roots that plays a major role in the culinary footprint of Pará. Before boiling, this juice is known as manipueira and contains linamarin, a toxic glycoside that can decompose to hydrogen cyanide. In this study, the cytotoxic and genotoxic effects of tucupi on cultured human lymphocytes were assessed using the comet assay and detection of apoptosis and necrosis by differential fluorescent staining with acridine orange-ethidium bromide. Tucupi concentrations (v/v) were determined using the methylthiazole tetrazolium biochemical test. Concentrations of tucupi that presented no genotoxic effects (2, 4, 8, and 16%) were used in our experiments. The results showed that under our study conditions, tucupi exerted no genotoxic effects; however, cytotoxic effects were observed with cell death mainly induced by necrosis. These effects may be related to the presence of hydrogen cyanide in the juice.

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Objetivou-se investigar a eficiência dos meios TCM 199 e PBS na conservação de folículos pré-antrais(FOPA) felinos in situ a 4ºC por 24 h. Foram utilizados 10 pares de ovários de gatas Felis catus, sendo a camadacortical fracionada e submetida a três diferentes tratamentos: controle, T1 (fragmentos conservados em TCM199) e T2 (fragmentos conservados em PBS). Realizou-se a preparação histológica do grupo controle, sendo T1e T2 mantidos a 4ºC, por 24 h, em caixa isotérmica. Ao final dos tratamentos, os FOPA foram classificados emprimordiais, primários e secundários, e os oócitos classificados em normais ou degenerados morfologicamente. Aavaliação histológica constatou que os FOPA do controle (63%) tiveram maior integridade morfológica (P > 0,05)quando comparados ao T2 (55%), sendo semelhantes ao T1 (59%). Não houve diferença (P < 0,05) entreT1(59%) e T2 (55%). Os folículos primordiais mantiveram maior integridade morfológica quando comparadosaos folículos em desenvolvimento, independentemente do tratamento. Com base nesses resultados, pode-seconcluir que é possível a manutenção da integridade morfológica de FOPA em ovários de gatas mantidos a 4ºCpor 24 h, utilizando o TCM 199 e que os folículos primordiais têm maior viabilidade, quando comparados aosfolículos em desenvolvimento.(AU)

The aim of this study was to investigate the efficiency of TCM 199 and PBS mediums in the conservationof feline preantral follicles (PAF) in situ at 4°C for 24 h. Ten pairs of ovaries of Felis catus cats were used, inwhich the cortical layer was fractioned and submitted to three different treatments: Control, T1 (fragmentspreserved in TCM 199) and T2 (fragments preserved in PBS). Histological preparation was carried out in thecontrol group, while T1 and T2 were kept at 4°C for 24 h in an isothermal box. After histological preparation ofControl, T1 and T2 the PAF were classified as primordial, primary and secondary and the morphology of theoocyte as normal and degenerated. Histological evaluation found that the PAF control group (63%) had a highermorphological integrity (P > 0.05) compared to T2 (55%), which was similar to T1 (59%). However, there wasno difference (P < 0.05) between T1 (59%) and T2 (55%) treatments. Primordial follicles maintained greatermorphological integrity when compared to developing follicles, regardless of the treatment. Based on theseresults, it can be concluded that it is possible to maintain the morphological integrity of PAF in ovaries of catstransported at 4°C for 24 h using TCM 199 and that primordial follicles have a higher viability compared todeveloping follicles.(AU)

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Objetivou-se investigar a eficiência dos meios TCM 199 e PBS na conservação de folículos pré-antrais(FOPA) felinos in situ a 4ºC por 24 h. Foram utilizados 10 pares de ovários de gatas Felis catus, sendo a camadacortical fracionada e submetida a três diferentes tratamentos: controle, T1 (fragmentos conservados em TCM199) e T2 (fragmentos conservados em PBS). Realizou-se a preparação histológica do grupo controle, sendo T1e T2 mantidos a 4ºC, por 24 h, em caixa isotérmica. Ao final dos tratamentos, os FOPA foram classificados emprimordiais, primários e secundários, e os oócitos classificados em normais ou degenerados morfologicamente. Aavaliação histológica constatou que os FOPA do controle (63%) tiveram maior integridade morfológica (P > 0,05)quando comparados ao T2 (55%), sendo semelhantes ao T1 (59%). Não houve diferença (P < 0,05) entreT1(59%) e T2 (55%). Os folículos primordiais mantiveram maior integridade morfológica quando comparadosaos folículos em desenvolvimento, independentemente do tratamento. Com base nesses resultados, pode-seconcluir que é possível a manutenção da integridade morfológica de FOPA em ovários de gatas mantidos a 4ºCpor 24 h, utilizando o TCM 199 e que os folículos primordiais têm maior viabilidade, quando comparados aosfolículos em desenvolvimento.

The aim of this study was to investigate the efficiency of TCM 199 and PBS mediums in the conservationof feline preantral follicles (PAF) in situ at 4°C for 24 h. Ten pairs of ovaries of Felis catus cats were used, inwhich the cortical layer was fractioned and submitted to three different treatments: Control, T1 (fragmentspreserved in TCM 199) and T2 (fragments preserved in PBS). Histological preparation was carried out in thecontrol group, while T1 and T2 were kept at 4°C for 24 h in an isothermal box. After histological preparation ofControl, T1 and T2 the PAF were classified as primordial, primary and secondary and the morphology of theoocyte as normal and degenerated. Histological evaluation found that the PAF control group (63%) had a highermorphological integrity (P > 0.05) compared to T2 (55%), which was similar to T1 (59%). However, there wasno difference (P < 0.05) between T1 (59%) and T2 (55%) treatments. Primordial follicles maintained greatermorphological integrity when compared to developing follicles, regardless of the treatment. Based on theseresults, it can be concluded that it is possible to maintain the morphological integrity of PAF in ovaries of catstransported at 4°C for 24 h using TCM 199 and that primordial follicles have a higher viability compared todeveloping follicles.

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The objective of this study was to determine the effect of different centrifugation forces in bovine sperm separation by discontinuous Percoll gradients for in vitro fertilization IVF. The semen samples from each bull were pooled or each bull were centrifuged separately and centrifuged in discontinuous Percoll gradients (30, 60 and 90%) at different forces: F1 (9000×g), F2 (6500×g), F3 (4500×g) and F4 (2200×g), according experiment. The sperm samples were evaluated to determine the concentration, motility, vigor, morphology, reactive oxygen species (ROS), integrity of the plasma membrane, lipid peroxidation, antioxidants and embryo development were also evaluated. No difference was observed in the concentration of sperm submitted to different centrifugation forces. The total percentage of motile sperm was increased after centrifugation at F3 and F4, and the ROS production at F1 was greater than the other forces. When the bulls semen were processed individually, no significant differences were observed for the sperm quality parameters between F1 and F4, including lipid peroxidation, antioxidants, cleavage rate and average time to the first cleavage. This work demonstrated for the first time that centrifugation at 2200×g enhanced the sperm penetration and fertilization rates without reducing sperm recovery compared to the typical centrifugation force (9000×g) currently used by the commercial bovine IVF industry.

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There is a constant search for new cancer treatments that are less aggressive and economically affordable. In this context, natural products extracted from plants, fungi, and microorganisms are of great interest. Pestheic acid, or dihidromaldoxin, is a chlorinated diphenylic ether extracted from the phytopathogenic fungus Pestalotiopsis guepinii (Amphisphaeriaceae). We assessed the cytotoxic, cytostatic, and genotoxic effects of pestheic acid in a gastric adenocarcinoma cell line (PG100). A decrease in clonogenic survival was observed. Pestheic acid also induced significant increases in both micronucleus and nucleoplasmic bridge frequency. However, we did not observe changes in cell cycle kinetics or apoptosis induction. Reactive oxygen species induced by diphenylic ethers may explain the genotoxicity and mutagenicity of pestheic acid. The absence of repair checkpoints that we observed is probably due to the fact that the PG100 cell line lacks the TP53 gene, which is common in gastric cancers. Even though pestheic acid has had a clear cytotoxic effect, the minimal inhibitory concentration was high, which shows that pestheic acid is not an active anticancer compound under these conditions.

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INTRODUCTION: Non-volatile oleoresins from neotropical Burseraceae are traditionally used for craft, technological and medicinal purposes. The crude resin is usually sold in popular markets of the forest communities. Adding value to this rainforest raw material requires establishing its composition. OBJECTIVE: To analyse the resin composition from different Burseraceae species and establish a minimally reproducible profile by gas chromatography, in order to parameterise its quality control. METHODOLOGY: Crude oleoresin samples of 10 Protium and Trattinnickia species and a commercial blend were subjected to hydrodistillation to remove volatile compounds. The chloroform-soluble residues were methylated, analysed by GC-FID (flame ionisation detection), and individual components were identified by analysing their mass fragmentation pattern in GC-MS and comparison with data from the literature. The blend solubility was assayed in 30 non-chlorinated solvents at three different proportions. RESULTS: The resins consisted exclusively of triterpenes, showing a common predominance of four major compounds in all the samples, corresponding to α-amyrin, ß-amyrin, α-amyrenone and ß-amyrenone. This profile was complemented with minor amounts of the tetracyclic ß-elemolic and α-elemolic acids, maniladiol, brein and other oxidised trace compounds. The better solvents for the resin were those chemically bearing more than four carbon atoms, as n-butyl acetate, 2-ethoxyethanol and isopropanol. The crude resin blend sold contained around 10% of insoluble material that was constituted by up to 70% inorganic residues mixed with humic acid derivatives, as attested by ash analysis and IR spectroscopy, respectively. CONCLUSION: The experimental results, complemented by a general inspection of the literature, demonstrated a systematically reproducible triterpene profile in Protium and Trattinnickia species.

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Trypanosoma cruzi is the epidemiological agent of Chagas' disease, affecting most of Central and South America, constituting a significant health and socio-economic problem. The parasite has a metabolism largely based on the consumption of amino acids, which participate in a diversity of metabolic pathways, leading to many crucial compounds for the survival of this parasite. Study of its enzymes has the potential to disclose new therapeutic targets and foster the development of new drugs. In this study, we employed computational approaches to reconstruct in silico the amino acid metabolic pathways of T. cruzi, aiming to link genomic information with functional information. For that, protein sequences from 570 EC classes belonging to 25 different pathways in general amino acid metabolism were downloaded from KEGG. A subset of 471 EC classes had at least one sequence deposited. Clustering of the proteins belonging to each EC class was performed using a similarity-based approach implemented in the tool AnEnPi. Reconstruction of the metabolic pathways comprising the amino acid metabolism of T. cruzi was performed by analyzing the output of BLASTP, using as query the dataset of predicted proteins of T. cruzi against all sequences of each individual cluster. This approach allowed us to identify 764 T. cruzi proteins probably involved in the metabolism of amino acids as well as the identification of several putative cases of analogy. Furthermore, we were able to identify several enzymatic activities of T. cruzi that were not previously included in KEGG.

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