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Germline de novo mutations can impact individual fitness, but their role in human male infertility is understudied. Trio-based exome sequencing identifies many new candidate genes affecting male fertility, including an essential regulator of male germ cell pre-mRNA splicing.

Abstract The translucency of dental ceramics [feldspathic ceramic (FC), lithium disilicate glass-ceramic (LD), zirconia-reinforced lithium silicate glass-ceramic (LS), and translucent zirconia (TZ)], in two thicknesses (1 and 2 mm), using different parameters was assessed. Translucency parameters ( TP ab *, TP00), contrast ratio (CR), light transmittance at 468 nm (T%), and light attenuation were determined. Data were analyzed by ANOVA, Tukey’s test, and Pearson correlations (α=0.05). CR ranged from 0.64 (LS-1 mm) to 0.96 (TZ-2 mm). TP ab * ranged from 3.7 (TZ-2 mm) to 32.2 (LS-1 mm). TP00 ranged from 3.0 (TZ-2 mm) to 26.4 (LS-1 mm). T% ranged from 14.9 (TZ-2 mm) to 75.2 (FC-1 mm). Light attenuation varied from 54% (LS-1 mm) to 84% (TZ-2 mm). There were significant strong negative correlations between CR and TP ab * (R=-0.997), CR and TP00 (R=-0.991), TP ab * and light attenuation (R=-0.888), and TP00 and light attenuation (R=-0.867). LS showed the highest translucency, whereas TZ showed the lowest translucency. When thickness increased, CR and light attenuation also increased, while TP and transmittance decreased.

Abstract Group A Streptococcus (Strep A) is a life-threatening human pathogen with no licensed vaccine. Here, we used a biopolymer particle (BP) approach to display repeats of Strep A vaccine candidate peptides p*17 and K4S2 derived from M and non-M protein, respectively. BPs densely displaying both peptides (BP-p*17-S2) were successfully assembled in one-step inside an engineered endotoxin-free Escherichia coli strain. Purified BP-p*17-S2 showed a spherical core-shell morphology with a biopolymer core and peptide shell. Upon formulation with aluminum hydroxide as adjuvant, BP-p*17-S2 exhibited a mean diameter of 2.9 µm and a positive surface charge of 22 mV. No cytotoxicity was detected when tested against HEK-293 cells. Stability studies showed that BP-p*17-S2 is ambient-temperature stable. Immunized mice showed no adverse reactions, while producing high titers of peptide specific antibodies and cytokines. This immune response could be correlated with protective immunity in an animal model of infection, i.e. intranasal challenge of mice with Strep A, where a significant reduction of >100-fold of pathogen burden in nose-associated lymphoid tissue, lung, and spleen was obtained. The cost-effective scalable manufacture of ambient-temperature stable BPs coated with Strep A peptides combined with their immunogenic properties offer an attractive alternative strategy to current Strep A vaccine development.

Streptococcus suis is a zoonotic pathogen affecting pigs and humans. This bacterium causes severe economic losses in the swine industry and poses a serious threat to public health and food safety. There is no effective commercial vaccine available for pigs or humans. In this study, we applied the biopolymer particle (BP) vaccine technology to incorporate seven conserved S. suis antigens (38 kDa protein (38), enolase (Enol), SSU1915, SSU1355, SSU0185, SSU1215, and SSU1773 (SSU1 and SSU2)). Two combinations of these antigens (38 and Enol; all SSU antigens designated as SSU1 and SSU2) were engineered to mediate production of BPs coated with either antigens 38 and Enol or SSU1 and SSU2 inside recombinant Escherichia coli. The isolated and purified empty BPs, 38-BP-Enol and SSU1-BP-SSU2, showed size ranges of 312–428 nm and 292–344 nm with and without the QuilA® adjuvant, respectively, and all showed a negative surface charge. Further characterization of purified BPs confirmed the presence of the expected antigen-comprising fusion proteins as assessed by tryptic peptide fingerprinting analysis using quadrupole time-of-flight mass spectrometry and immunoblotting. Vaccination with 38-BP-Enol and SSU1-BP-SSU2 formulated with and without QuilA® adjuvant induced significant antigen-specific humoral immune responses in mice. Antigen-coated BPs induced significant and specific Ig (IgM + IgG) and IgG immune responses (1.0 × 106–1.0 × 107) when compared with mice vaccinated with empty BPs. Functionality of the immune response was confirmed in challenge experiments using an acute murine S. suis infection model, which showed 100% survival of the 38-BP-Enol and SSU1-BP-SSU2 vaccinated mice compared to 70% survival when vaccinated with empty BPs. Overall, our data suggest that S. suis antigen-coated BPs could be developed into particulate vaccines that induce protective immunity against S. suis infections.

Despite numerous efforts to develop an effective vaccine against Pseudomonas aeruginosa, no vaccine has yet been approved for human use. This study investigates the utility of the P. aeruginosa inherently produced polyhydroxyalkanaote (PHA) inclusions and associated host–cell proteins (HCP) as a particulate vaccine platform. We further engineered PHA inclusions to display epitopes derived from the outer membrane proteins OprF/OprI/AlgE (Ag) or the type III secretion system translocator PopB. PHA and engineered PHA beads induced antigen-specific humoral, cell-mediated immune responses, anti-HCP and anti-polysaccharide Psl responses in mice. Antibodies mediated opsonophagocytic killing and serotype-independent protective immunity as shown by 100% survival upon challenge with P. aeruginosa in an acute pneumonia murine model. Vaccines were stable at 4 °C for at least one year. Overall, our data suggest that vaccination with subcellular empty PHA beads was sufficient to elicit multiple immune effectors that can prevent P. aeruginosa infection.

Abstract Background Stagnant flooding, where water of 25–50 cm remains until harvest time, is a major problem in rainfed lowland areas. Most of the Sub1 varieties, which can withstand around 2 weeks of complete submergence, perform poorly in these conditions. Hence, varieties tolerant of stagnant flooding are essential. Results This paper presents the first study to map QTLs associated with tolerance to stagnant flooding, along with a parallel study under normal irrigation, using an F7 mapping population consisting of 148 RILs derived from a cross of Ciherang-Sub1 and the stagnant-flooding tolerant line IR10F365. Phenotypic data was collected for 15 key traits under both environments. Additionally, survival rate was measured under stress conditions. Genotyping was performed using the Illumina Infinium genotyping platform with a 6 K SNP chip, resulting in 469 polymorphic SNPs. Under stress and irrigated conditions, 38 and 46 QTLs were identified, respectively. Clusters of QTLs were detected in both stress and normal conditions, especially on chromosomes 3 and 5. Conclusions Unique and common QTLs were identified and their physiological consequences are discussed. These beneficial QTLs can be used as targets for molecular breeding and can be further investigated to understand the underlying molecular mechanisms involved in stagnant flooding tolerance in rice.

Objectives: To determine the prevalence of increased serum fibrinogen levels and its association with cardiometabolic risk factors in overweight or obese children and adolescents. Methods: Cross‐sectional study with 138 children and adolescents (overweight or obese) followed at a reference outpatient clinic of the public health care network. Fibrinogen concentration was divided into quartiles, and values above or equal to the third quartile were considered high. The association between high fibrinogen values and cardiometabolic risk factors was assessed using Pearson's chi‐squared test or Fisher's exact test, as necessary. Logistic regression was used to adjust variables predictive of fibrinogen levels. Analyses were performed using SPSS version 22.0 and SAS software, considering a confidence interval of 95%. Results: Serum fibrinogen levels were elevated in 28.3% of individuals, showing association with the presence of high CRP (p = 0.003, PR: 2.41, 95% CI: 1.30‐4.46) and the presence of four or more risk factors (p = 0.042; PR: 1.78, 95% CI: 1.00‐3.17). After a logistic regression, only elevated CRP remained associated with altered fibrinogen levels (p = 0.024; PR: 1.32; 95% CI: 1.09‐5.25). Conclusions: Increased fibrinogen was prevalent in the study population and was associated with ultrasensitive C‐reactive protein and the presence of four or more cardiovascular risk factors; it should be included in the assessment of individuals at risk.

OBJETIVO: Verificar a relação entre a concentração de ácido úrico sérico de acordo com a presença ou não de esteatose hepática não alcoólica e/ou síndrome metabólica (SM) em crianças e adolescentes com sobrepeso ou obesidade. MÉTODOS: Estudo transversal desenvolvido no período de abril/2009 a março/2010, incluindo 129 crianças e adolescentes atendidos no Centro de Obesidade Infantil. Foi realizada antropometria, aferição da pressão arterial, dosagem dos exames laboratoriais e o diagnóstico de esteatose hepática por exame ultrassonográfico. Para o diagnóstico de SM, foram utilizados os critérios da National Cholesterol Education Program/Adult Treatment Panel III adaptados para faixa etária. Para avaliação da associação do ácido úrico com os grupos, foi realizado o teste do Qui-quadrado ou Fisher, adotando-se o intervalo de confiança de 95%. Para comparação de médias, utilizou-se o ANOVA One Way. Para o ajuste das variáveis foi utilizada a regressão logística múltipla. Os dados foram processados no SPSS versão 17. RESULTADOS: Níveis elevados de ácido úrico associaram-se significativamente à adolescência, SM e pressão arterial sistólica. O maior quartil de ácido úrico apresentou valores médios significativamente mais elevados de índice de massa corpórea, circunferência abdominal, pressão arterial sistólica, pressão arterial diastólica, triglicerídeos, colesterol total e HOMA-IR, e menor média do colesterol HDL. No modelo final só permaneceram associadas aos níveis de ácido úrico a faixa etária e a presença de síndrome metabólica. CONCLUSÕES: Níveis elevados de ácido úrico estiveram associados à síndrome metabólica e à adolescência, o que não foi observado com a esteatose hepática.

Objective: To investigate the association between serum uric acid concentration according to the presence or absence of non-alcoholic fatty liver disease (NAFLD) and/or metabolic syndrome (MS) in overweight or obese children and adolescents. Methods: This was a cross-sectional study conducted from April of 2009 to March of 2010, including 129 children and adolescents treated at the Center for Childhood Obesity. Anthropometric data, blood pressure measurements, and laboratory test results were obtained, and NAFLD diagnosis was made by ultrasound. The diagnosis of MS was made using the criteria of the National Cholesterol Education Program/Adult Treatment Panel III, adapted to age range. The chi-squared test or or Fisher's test were used to evaluate the association of uric acid with the groups, with a 95% confidence interval. One- way analysis of variance (ANOVA) was used for comparison of means. Multiple logistic regression was used for adjustment of variables. The data were analyzed with the Statistical Package for Social Sciences (SPSS), release 17. Results: High levels of uric acid were significantly associated with adolescence, MS, and systolic blood pressure. The highest quartile of uric acid showed significantly higher values of body mass index, waist circumference, systolic blood pressure, diastolic blood pressure, triglycerides, total cholesterol, and homeostatic model assessment index (HOMA-IR), and lower mean values of HDL cholesterol. In the final model, only age range and the presence of MS remained associated with uric acid levels. Conclusions: High levels of uric acid were associated with MS and adolescence, which was not observed with NAFLD.

Verificou-se o efeito do acondicionamento em saliva artificial na resistência à flexão e no tempo de vida de uma porcelana feldspática dentária. Os espécimes na forma de disco foram preparados por sinterização, usinagem e polimento e ensaiados pelo método da fadiga dinâmica para determinação dos parâmetros de crescimento de trinca subcrítico (SCG), com taxas de tensão de 0,01 a 100 MPa/s em meio de saliva artificial a 37 °C. Antes do ensaio de flexão biaxial, parte dos espécimes foi imersa em saliva artificial a 37 °C por 10 dias e outra parte foi mantida ao ar. O acondicionamento em saliva causou aumento de ~11% na resistência à flexão biaxial; aumento de 20% no módulo de Weibull; aumento de 20% no coeficiente de susceptibilidade ao fenômeno de SCG; aumento de 50% na resistência à flexão prevista no tempo de fratura de 1 ano; manutenção da tenacidade à fratura; e diminuição de 5% da dureza superficial. Os resultados indicaram que, ao menos no caso investigado, a exposição da porcelana à saliva não degrada as propriedades mecânicas e pode até aumentar a resistência à flexão e à propagação lenta de trincas.The effects of artificial saliva storage on flexural strength and lifetime prediction of a dental feldspathic porcelain were investigated. The disk-shaped specimens were sintered, machined and polished and then subjected to a dynamic fatigue test in order to determine the slow crack growth (SCG) parameters, by means of biaxial flexure tests at stress rates of 0.01 to 100 MPa/s. The specimens remained immersed in artificial saliva at 37 ºC during the whole test. Before the biaxial flexure strength test, some specimens were stored in artificial saliva at 37 ºC for 10 days while others were stored in air at ambient temperature. The specimens stored in artificial saliva presented an ~11% increase in biaxial flexure strength; a 20% increase in Weibull modulus; a 20% increase in SCG susceptibility coefficient; a 50% increase in the predicted flexural strength for a lifetime of 1 year, a 5% decrease in superficial hardness. Similar fracture toughness values were determined for groups stored in air and in artificial saliva. The results indicated that, at least in the present study, storage of a dental porcelain in artificial saliva did not decrease the mechanical properties investigated and can even increase the flexural strength value and the resistance to slow crack growth phenomenon.