Acute murine-betacoronavirus infection impairs testicular steroidogenesis and the quality of sperm production.

Although several testicular alterations promoted by coronavirus infection have been demonstrated, the extent, causes, and players of testicular pathogenesis are not totally understood. The present study aimed to investigate the short-term effects on male fertility of intranasally administered murine hepatitis virus strain 3 (MHV-3), a member of the genus Betacoronavirus, which causes a severe systemic acute infection. This mouse model might be used as a in vivo prototype for investigating the impact of betacoronavirus on the endocrine and exocrine testicular functions with the advantage to be performed in a biosafety level 2 condition. Herein, we performed virological, histopathological, and molecular studies regarding the testicular spermatogenesis and the spermatic quality analyses in an MHV-3-infected C57BL/6 mice. The main outcomes showed that MHV-3 infects mouse testis and induces a testicular inflammatory state, impairing the steroidogenic pathway. The infection led to several alterations in the testicular parenchyma, such as: seminiferous epithelium sloughing, retention of residual bodies, germ cell apoptosis, alterations in intercellular junction proteins, and worse spermatogenic parameters. Moreover, the levels of plasmatic testosterone as well as the quality of sperm production reduced. Therefore, the present data suggest that the viral/inflammatory impairment of the steroidogenic pathway and the consequent imbalance of androgen levels is critical in testicular pathology, disturbing the SC barrier function and the germ cell differentiation. Our study is important for comprehending the effects of beta coronavirus infections on testis function in order to develop treatments that could prevent virus-mediated male infertility.

Lactobacillus delbrueckii UFV-H2b20 increases IFN-γ production and CD39+CD73+ Treg cell numbers in lungs, and protects mice against experimental allergic asthma.

Asthma is a disorder characterized by airflow obstruction, inflammation, declining airway function, bronchial hyperresponsiveness and tissue remodelling. Probiotics are defined as "live microorganisms that, when administered in adequate amounts, confer a health benefit on the host". The use of probiotics is becoming increasingly studied and recent evidence has suggested that it may provide therapeutic benefits in asthma and other diseases. Lactobacillus delbrueckii UFV-H2b20 fulfils all the requirements to be classified as probiotic. Previous studies have already shown the ability of L. delbrueckii UFV-H2b20 to stimulate the immune system. Our objective was to evaluate the protective effects of L. delbrueckii UFV-H2b20 in experimental allergic asthma. We used a murine model of ovalbumin-induced allergic airway inflammation to mimic allergic asthma. Oral treatment with L. delbrueckii UFV-H2b20 improves respiratory parameters and inhibits the inflammatory response in the lungs by decreasing the numbers of inflammatory monocytes, eosinophils and alveolar macrophages, as well as IgE levels. Treatment increased the IFN-γ/IL-4 cytokine ratio. Levels of IL-10 in the lungs were also increased in treated animals. Our results also showed that the probiotic administration increases the number of CD39+CD73+ T regulatory lymphocytes in the lung, suggesting a role for purinergic signals in the regulation of inflammation promoted by the treatment. Understanding the mechanisms of modulation of the immune system by probiotics could allow the development of probiotic preparations that are safe and have a direct action. Our results suggest that oral administration of L. delbrueckii UFV-H2b20 could be helpful to treat chronic inflammatory airway diseases, such as asthma.

Optical Property Stability of Light-Cured versus Precured CAD-CAM Composites.

Objective: The aim of this study was to evaluate the optical property changes after staining of precured (PC) and light-cured (LC) composites. Materials and Methods: Specimens were prepared using different LC composites (GrandioSO-Voco, Filtek Z350-3M/ESPE, Opallis-FGM, and Kalore-GC) and four PC blocks (Grandio Blocs-Voco, Lava Ultimate-3M ESPE, Brava Block-FGM, and Cerasmart-GC) from the same manufacturers (n = 20). Baseline color, gloss, translucency, and fluorescence were evaluated. The staining protocol was performed for 15 days, and the final optical properties were reevaluated. Results: The changes in each property were calculated (ΔGloss, ΔTranslucency, ΔFluorescency, ΔE ∗ 00). Data were analyzed by ANOVA and Tukey's test (α = 5%). Changes in all properties were observed after staining for all materials, with darkening and reduction of gloss, fluorescence, and translucency. Nonsignificant differences were observed between the light-cured and precured materials of the same manufacturer for ΔG and ΔT, but significant differences existed for ΔF and ΔE ∗ 00. For ΔF, the only significant differences were observed between Brava Block and Opallis (smaller). For ΔE ∗ 00, only the light-cured composites GrandioSO and Z350 showed significantly less change than the corresponding blocks. Precured composites were affected the same way as light-cured ones by the staining in relation to the reduction of gloss and translucency. Conclusion: A higher reduction in fluorescence was observed for only one brand of block and was similar for the others. The two brands of light-cured materials showed less staining, while for the others, the staining was similar. The effects of staining vary according to the composite formulation.

Germination of small tropical seeds has distinct light quality and temperature requirements, depending on microhabitat.

The coexistence of plant species in tropical rainforests is related to specific abiotic resources, varying according to the occurrence microhabitat of each species. Light quality is the main abiotic factor influencing germination of small seeds; however, studies often do not discriminate its effect from that of light irradiance. This study compared specific requirements for seed germination of ten small-seeded species, with restricted occurrence in only one of three contrasting microhabitats: forest understorey, edge of clearings and open areas. Laboratory experiments were carried out to test temperature regime (constant or fluctuating), light quality (R:FR) and light irradiance (PAR), which reproduce high and low conditions commonly found in the microhabitats. Seed germination of all species occurred between 20 and 30 °C, only seeds of open area species were able to germinate at 35 °C and no species required alternating temperatures to germinate. Irrespective of species and microhabitat, a decrease in the R:FR reduced the germination percentage; however, there were differences in the capacity to germinate at low R:FR. The values of R:FR50% were higher for open area and edge species (0.441-0.345) than for understorey species (0.181-0.109), with few exceptions. For all species and most of the tests, germination was not influenced by PAR. Light quality is the most important light signal for germination of small seeds; irradiance has little effect. Our results suggest two distinct patterns of germination for small-seeded species: open area and edge species are light-demanding and require high R:FR to germinate, while understorey species are shade-tolerant and germinate at low R:FR. These differences are responsible for distinct microhabitat occurrence and help to explain the coexistence of species in tropical forests.

Germination, viability and dormancy of 47 species from threatened tropical montane grassland in southeast Brazil: Implications for ex situ conservation.

To mitigate anthropogenic impacts on plant diversity in tropical montane grasslands, one of the most threatened ecosystems in Brazil, it will be essential to develop ex situ conservation strategies to preserve wild species. The lack of basic research on the seed storage behaviour of grassland species may, however, limit their use for reintroduction and restoration projects. We investigated seed storage behaviour at the community level by comparing the effects of cold-low RH (10 °C; 10% RH) and freezing-low RH (20 °C; 10% RH) conditions on seed viability, germination and dormancy of 47 species. Fresh seeds of 43% of the species showed primary dormancy. More than half of the species showed high seed survival responses (viability >60%) under both storage temperatures. Despite a variety of dormancy responses among the different species, the low RH storage conditions tested released dormancy for most species during 12- and 30-month storage times. Multivariate analysis of the best (freezing-low RH, 30 months) storage condition evidenced the formation of five distinct groups, three with species having high conservation potential in seed banks. Although further studies are needed to test dormancy-breaking treatments and improve seed conservation practices, this first approach to assessing seed banking techniques could contribute to demand for locally adapted seeds for ecological restoration projects in tropical montane grasslands.

Evaluating the menopausal transition with the STRAW + 10 in a Brazilian cohort of women with HIV, 2015-2016.

BACKGROUND: Menopausal transition is a physiological process encompassing hormonal and body changes that impact women's health and life quality. This period may be characterized by the Stages of Reproductive Aging Workshop (STRAW + 10) criteria using menstrual patterns. Use of the STRAW + 10 is uncertain in HIV infection. We aimed to characterize menopausal transition in women with HIV (WWH) using the STRAW + 10 criteria, hormonal measures and menopause symptoms. METHODS: We performed a cross-sectional study, nested to the HIV-Infected Women's Cohort, in Rio de Janeiro, Brazil. Eligible women included those aged 30 years or older, without clinical or surgical menopause, hormonal contraception, replacement therapy and ovarian disorders. We conducted face-to-face interviews and collected blood samples for follicle stimulating hormone (FSH) and estradiol measures. RESULTS: We enrolled 328 WWH (28.3% of women in the cohort). The distribution of age, hormonal levels and reported symptoms per each STRAW + 10 stage was consistent with the expected distribution in the menopausal transition. Age and FSH significantly increased and estradiol decreased from stage -2 (7 + days of menstrual delay) to stage +2 (8 + years of amenorrhea). CONCLUSIONS: The present results support use of the STRAW + 10 to characterize the menopausal transition of WWH with good clinical and immunological control.

Food consumption on campus is associated with meal eating patterns among college students.

This is a cross-sectional analysis of data obtained in the baseline of the Longitudinal Study on the Lifestyle and Health of University Students (n 685) carried out in a public Brazilian university. Food intake was assessed using a 24-h dietary recall. Dietary patterns (DP) for breakfast, lunch and dinner were identified using principal component analysis. Generalised linear models were used to analyse the variables associated with each DP. Three DP were extracted for each meal: breakfast: 'White bread and butter/margarine', 'Coffee and tea' and 'Sausages, whole wheat bread and cheese'; lunch: 'Traditional', 'Western' and 'Vegetarian' and dinner: 'Beans, rice and processed juice', 'White bread and butter/margarine' and 'White meat, eggs and natural juice'. Students who had meals at the campus showed greater adherence to the 'White bread and butter/margarine' (exp (ßadj) = 1·15, 95 % CI 1·11, 1·19) and 'Coffee and tea' (exp (ßadj) = 1·06, 95 % CI 1·02, 1·10) breakfast patterns; 'Western' lunch pattern (exp (ßadj) = 1·04, 95 % CI 1·01, 1·08) and to the 'Beans, rice and processed juice' dinner pattern (exp (ßadj) = 1·10, 95 % CI 1·06, 1·14). Having meals at the campus was associated with lower adherence to the 'Sausages, whole wheat bread and cheese' breakfast pattern (exp (ßadj) = 0·93, 95 % CI 0·89, 0·97), 'Traditional' lunch pattern (exp (ßadj) = 0·96, 95 % CI 0·93, 0·99) and to the 'White bread and butter/margarine' (exp (ßadj) = 0·96, 95 % CI 0·93, 0·99) and 'White meat, eggs and natural juice' (exp (ßadj) = 0·96, 95 % CI 0·93, 0·99) dinner pattern. The food environment at campus may influence students' DP. Recognising meal eating patterns is important to support healthy eating promotion strategies on campus. Adjustments in the University Canteen menu could contribute to healthier eating choices among students.

Quantification of bleeding during dental extraction in patients on dual antiplatelet therapy.

In spite of the possibility of triggering thromboembolic events, many professionals indicate the suspension of antiplatelet agents before dental surgical procedures. The aim of this study was to perform a quantitative assessment of intraoperative bleeding in patients on dual antiplatelet therapy. A case-control study was conducted in patients on dual antiplatelet therapy (APT group) and in patients who did not use these medications (control group). The following examinations were requested: complete blood cell count, blood coagulation tests, and platelet aggregation. The quantity of bleeding was measured intraoperatively by collection of aspirated blood. The mean volume of blood lost during the surgical procedure was 6.10ml in the control group and 16.07ml in the APT group (P=0.002). The mean volume of blood lost per minute was 0.60ml/min in the control group and 1ml/min in the APT group (P=0.001), with local haemostatic methods being sufficient to control the bleeding. There was no postoperative bleeding complication in any case. Patients on dual antiplatelet therapy presented a larger volume of bleeding, but this could be controlled by means of local haemostatic measures. Therefore, there is no need to stop either of the two dual antiplatelet therapy medications before dental extractions.

Daily ingestion of the probiotic Lactobacillus paracasei ST11 decreases Vaccinia virus dissemination and lethality in a mouse model.

Vaccinia virus (VACV) is an important pathogen. Although studies have shown relationships between probiotics and viruses, the effect of probiotics on VACV infection is unknown. Therefore, this work aims to investigate the probiotics effects on VACV infection. Mice were divided into four groups, two non-infected groups, one receiving the probiotic, the other one not receiving it, and two groups infected intranasally with VACV Western Reserve (VACV-WR) receiving or not receiving the probiotic. Viral titres in organs and cytokine production in the lungs were analysed. Lung samples were also subjected to histological analysis. The intake of probiotic results in reduction in viral spread with a significant decrease of VACV titer on lung, liver and brain of treated group. In addition,treatment with the probiotic results in attenuated mice lung inflammation showing fewer lesions on histological findings and decreased lethality in mice infected with VACV. The ingestion of Lactobacillus paracasei ST11 (LPST11) after VACV infection resulted in 2/9 animal lethality compared with 4/9 in the VACV group. This is the first study on probiotics and VACV interactions, providing not only information about this interaction, but also proposing a model for future studies involving probiotics and other poxvirus.

Quantification of bleeding during dental extraction in patients on dual antiplatelet therapy

In spite of the possibility of triggering thromboembolic events, many professionals indicate the suspension of antiplatelet agents before dental surgical procedures. The aim of this study was to perform a quantitative assessment of intraoperative bleeding in patients on dual antiplatelet therapy. A case-control study was conducted in patients on dual antiplatelet therapy (APT group) and in patients who did not use these medications (control group). The following examinations were requested: complete blood cell count, blood coagulation tests, and platelet aggregation. The quantity of bleeding was measured intraoperatively by collection of aspirated blood. The mean volume of blood lost during the surgical procedure was 6.10ml in the control group and 16.07ml in the APT group (P=0.002). The mean volume of blood lost per minute was 0.60ml/min in the control group and 1ml/min in the APT group (P=0.001)...

Handgrip Strength Cutoff Points to Identify Mobility Limitation in Community-dwelling Older People and Associated Factors.

BACKGROUND: Sarcopenia is defined as a progressive and generalized loss of skeletal muscle mass and strength. The specific threshold of muscle weakness that leads to mobility limitations has not been identified. OBJECTIVES: To determine the best cutoff point of handgrip strength for identifying mobility limitation and to investigate the factors associated with muscle weakness and mobility limitation in community-dwelling older people. DESIGN: Transversal study. SETTING: Cities of Belo Horizonte, Barueri and Santa Cruz in Brazil. PARTICIPANTS: 1374 community-dwelling older people from the Frailty study in Brazilian older people (FIBRA Study). MEASUREMENTS: Outcomes included muscle weakness determined according to gender-specific handgrip strength cutoff points generated by Receiver Operating Characteristic curves, mobility limitation defined as a gait speed ≤ 0.8 m/s; and a combination of both muscle weakness and mobility limitation. Associated factors included socio-demographic variables, lifestyle, anthropometrics, health conditions, use of health services and disability. RESULTS: The cutoff points of handgrip strength with the best balancing between sensitivity and specificity for mobility limitation were 25.8 kgf for men (sensitivity 69%, specificity 73%) and 17.4 kgf (sensitivity 60%, specificity 66%) for women. Age and disability in instrumental activities of daily living were associated with all outcomes. Women had greater odds of mobility limitation than men. Physical inactivity, body fat, diabetes, depression, sleeping disturbances, number of medications and occurrence of falls remained as significant associated factors in the final model. CONCLUSIONS: Handgrip strength can be a useful tool to identify mobility limitation in clinical practice. Interventions to prevent or minimize impacts of sarcopenia should stimulate physical activity and improvement of body composition in addition to the management of chronic diseases and disabilities.

Short term inhalation toxicity of a liquid aerosol of glutaraldehyde-coated CdS/Cd(OH)2 core shell quantum dots in rats.

Quantum dots exhibit extraordinary optical and mechanical properties, and the number of their applications is increasing. In order to investigate a possible effect of coating on the inhalation toxicity of previously tested non-coated CdS/Cd(OH)2 quantum dots and translocation of these very small particles from the lungs, rats were exposed to coated quantum dots or CdCl2 aerosol (since Cd(2+) was present as impurity), 6h/d for 5 consecutive days. Cd content was determined in organs and excreta after the end of exposure and three weeks thereafter. Toxicity was determined by examination of broncho-alveolar lavage fluid and microscopic evaluation of the entire respiratory tract. There was no evidence for translocation of particles from the respiratory tract. Evidence of a minimal inflammatory process was observed by examination of broncho-alveolar lavage fluid. Microscopically, minimal to mild epithelial alteration was seen in the larynx. The effects observed with coated quantum dots, non-coated quantum dots and CdCl2 were comparable, indicating that quantum dots elicited no significant effects beyond the toxicity of the Cd(2+) ion itself. Compared to other compounds with larger particle size tested at similarly low concentrations, quantum dots caused much less pronounced toxicological effects. Therefore, the present data show that small particle sizes with corresponding high surfaces are not the only factor triggering the toxic response or translocation.

Identification of expressed sequences in the coffee genome potentially associated with somatic embryogenesis.

Brazil possesses the most modern and productive coffee growing farms in the world, but technological development is desired to cope with the increasing world demand. One way to increase Brazilian coffee growing productivity is wide scale production of clones with superior genotypes, which can be obtained with in vitro propagation technique, or from tissue culture. These procedures can generate thousands of clones. However, the methodologies for in vitro cultivation are genotype-dependent, which leads to an almost empirical development of specific protocols for each species. Therefore, molecular markers linked to the biochemical events of somatic embryogenesis would greatly facilitate the development of such protocols. In this context, sequences potentially involved in embryogenesis processes in the coffee plant were identified in silico from libraries generated by the Brazilian Coffee Genome Project. Through these in silico analyses, we identified 15 EST-contigs related to the embryogenesis process. Among these, 5 EST-contigs (3605, 9850, 13686, 17240, and 17265) could readily be associated with plant embryogenesis. Sequence analysis of EST-contig 3605, 9850, and 17265 revealed similarity to a polygalacturonase, to a cysteine-proteinase, and to an allergenine, respectively. Results also show that EST-contig 17265 sequences presented similarity to an expansin. Finally, analysis of EST-contig 17240 revealed similarity to a protein of unknown function, but it grouped in the similarity dendrogram with the WUSCHEL transcription factor. The data suggest that these EST-contigs are related to the embryogenic process and have potential as molecular markers to increase methodological efficiency in obtaining coffee plant embryogenic materials.

Construction and characterization of a BAC library from the Coffea arabica genotype Timor Hybrid CIFC 832/2.

Most of the world's coffee production originates from Coffea arabica, an allotetraploid species with low genetic diversity and for which few genomic resources are available. Genomic libraries with large DNA fragment inserts are useful tools for the study of plant genomes, including the production of physical maps, integration studies of physical and genetic maps, genome structure analysis and gene isolation by positional cloning. Here, we report the construction and characterization of a Bacterial Artificial Chromosome (BAC) library from C. arabica Timor Hybrid CIFC 832/2, a parental genotype for several modern coffee cultivars. The BAC library consists of 56,832 clones with an average insert size of 118 kb, which represents a dihaploid genome coverage of five to sixfold. The content of organellar DNA was estimated at 1.04 and 0.5 % for chloroplast and mitochondrial DNA, respectively. The BAC library was screened for the NADPH-dependent mannose-6-phosphate reductase gene (CaM6PR) with markers positioned on four linkage groups of a partial C. arabica genetic map. A mixed approach using PCR and membrane hybridization of BAC pools allowed for the discovery of nine BAC clones with the CaM6PR gene and 53 BAC clones that were anchored to the genetic map with simple sequence repeat markers. This library will be a useful tool for future studies on comparative genomics and the identification of genes and regulatory elements controlling major traits in this economically important crop species.

A minimal cytomegalovirus intron A variant can improve transgene expression in different mammalian cell lines.

The expression enhancement by cytomegalovirus promoter and different intron A (IA) variants were evaluated in CHO-K1, HepG2, HEK-293 and COS-7 cells by assessing the levels of luciferase activity. This data along with mRNA levels measurement indicated that the construct harboring an IA variant with a 200-nucleotide deletion (Δ200) had the greatest impact on increasing luciferase expression among all constructs evaluated. Based on these results, we redesigned pCMV-IA variants and cloned them into plasmids expressing a humanized antibody. These plasmids were then used to transfect CHO-K1 cells. Production of the antibody was not augmented with the Δ200 promoter variant. The 600-nucleotide deletion (Δ600) and whole IA promoter variants expressed similar levels of the recombinant protein. These data indicate that the IA-based enhanced expression of transgenes depends on a small region within the intron.

Cervical cytological abnormalities and factors associated with high-grade squamous intraepithelial lesions among HIV-infected women from Rio de Janeiro, Brazil.

Although cervical cancer remains a major public health problem in Brazil, knowledge of cervical cytological abnormalities among HIV-infected women remains scarce. At baseline evaluation of a cohort followed in Rio de Janeiro, Brazil, 703 HIV-infected women underwent cytology-based cervical cancer screening and human papillomavirus (HPV) DNA testing. Poisson regression analysis was used to evaluate the association of factors with the presence of high-grade squamous intraepithelial lesions (HSIL). Cervical cytology was abnormal in 24.3% of the women; 4.1% had HSIL. Beyond HPV infection, factors independently associated with the presence of HSIL was age (≥25 and ≤40 years, prevalence ratio [PR] 2.60, 95% confidence interval [CI] 1.11-6.10), and more than three pregnancies was protective (PR 0.33, 95% CI 0.11-0.94). High coverage of cervical cancer screening is warranted to prevent morbidity and mortality from cervical cancer in this population.

Quantitative electroencephalography power and coherence measurements in the diagnosis of mild and moderate Alzheimer's disease / Medidas de coerência e de potência absoluta no eletrencefalograma quantitativo no diagnóstico da doença de Alzheimer

OBJECTIVE: To evaluate the contribution of quantitative electroencephalographic (qEEG) analyses in the diagnosis of Alzheimer's disease (AD). METHOD: Thirty-five patients from the Neurology Outpatients Clinic of PUC-Campinas, diagnosed with AD according to the NINCDS/ADRDA were evaluated, and compared with a control group consisting of 30 individuals with no cognitive deficit. The procedures consisted of clinical-neurological, cognitive and behavioral analyses and the qEEG (absolute power and coherence). RESULTS: The AD group presented greater absolute power values in the delta and theta bands, greater theta/alpha indices and less frontal alpha and beta coherence. Logistic multiple regression models were constructed and those only showing variations in the qEEG (frontal alpha coherence and left frontal absolute theta power) showed an accuracy classification (72.3 percent) below that obtained in the mini-mental state examination (93 percent). CONCLUSION: The study of coherence and power in the qEEG showed a relatively limited accuracy with respect to its application in routine clinical practice.

OBJETIVO: Avaliar a contribuição das análises quantitativas do eletroencefalograma (qEEG) no diagnóstico da doença de Alzheimer (DA). MÉTODO: Foram avaliados 35 pacientes do ambulatório de Neurologia Clínica da PUC-Campinas, com o diagnóstico de DA segundo o NINCDS/ADRDA e comparados a 30 indivíduos, sem déficit cognitivo, de grupo controle. Os procedimentos foram avaliação clínico-neurológica, cognitiva e comportamental e EEGq (potência absoluta e coerência). RESULTADOS: O grupo DA apresentou maiores potências absolutas nas faixas delta e teta, maiores índices teta/alfa e menor coerência alfa e beta frontal. Foram construídos modelos de regressão múltipla logística e aquele que contou apenas com variáveis do EEGq (coerência alfa frontal e potência absoluta teta frontal esquerda) teve acurácia de classificação (72,3 por cento), inferior à obtida com o mini-exame do estado mental (93 por cento). CONCLUSÃO: O estudo de coerência e potência no qEEG tem acurácia relativamente limitada no sentido de aplicação prática clínica rotineira.

Spidroins from the Brazilian spider Nephilengys cruentata (Araneae: Nephilidae).

Spiders produce up to six different kinds of silk, each one for a specific biological function. Spider silks are also known for their unique mechanical properties. The possibility of producing new materials with similar properties motivated research on these silk proteins (spidroins). Using expression sequence tags, we identified four spidroins produced by major ampullate, minor ampullate, flagelliform and tubuliform silk glands from the Brazilian spider Nephilengys cruentata (Araneae: Nephilidae). The new protein sequences showed substantial similarity to other spidroins previously described, with high content of alanine and glycine due to the presence of the highly repetitive motifs (polyAla, (GA)n, (GGX)n, (GPGGX)n). Similarities among sequences were also observed between the different spidroins with the exception of tubuliform spidroin, which presents a unique complex amino acid sequence with high amounts of serine and low amounts of glycine.