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Background: Culex quinquefasciatus has a widespread distribution across tropical and sub-tropical regions, and plays an important role in the transmission of vector-borne diseases of public health importance, including lymphatic filariasis (LF) and multiple arboviruses. Increased resistance to insecticides threatens the efficacy and sustainability of insecticide-based anti-vector interventions which mitigate the burden of mosquito transmitted diseases in endemic regions. In C. quinquefasciatus two non-synonymous voltage gated sodium channel (Vgsc) variants, both resulting in a leucine to phenylalanine change at codon 1014, are associated with resistance to pyrethroids and DDT. This tri-allelic variation has compromised the ability to perform high-throughput single-assay screening. To facilitate the detection and monitoring of the Vgsc-1014 locus in field-caught mosquitoes, an Engineered-Tail Allele-Specific-PCR (ETAS-PCR) diagnostic assay was developed and applied to wild mosquitoes from Brazil, Tanzania and Uganda., Results: This new cost-effective, single-tube assay was compared to two, well-established, genotyping approaches, pyrosequencing and TaqMan. The ETAS-PCR assay showed high specificity for discriminating the three alleles at Vgsc-L1014F, with genotyping results strongly correlated with pyrosequencing and TaqMan results (98.64% and 100% respectively)., Conclusions: Our results support the utility of the ETAS-PCR/Vgsc-1014 diagnostic assay, which stands as an effective alternative for genotyping tri-allelic variants.

Our objective was to evaluate changes in breeding values for carcass traits of two meat-type quail (Coturnix coturnix) strains (LF1 and LF2) to changes in the dietary (methionine + cystine):lysine ([Met + Cys]:Lys) ratio due to genotype by environment (G × E) interaction via reaction norm. A total of 7000 records of carcass weight and yield were used for analyses. During the initial phase (from hatching to day 21), five diets with increasing (Met + Cys):Lys ratios (0.61, 0.66, 0.71, 0.76 and 0.81), containing 26.1% crude protein and 2900 kcal ME/kg, were evaluated. Analyses were performed using random regression models that included linear functions of sex (fixed effect) and breeding value (random effect) for carcass weight and yield, without and with heterogeneous residual variance adjustment. Both fixed and random effects were modelled using Legendre polynomials of second order. Genetic variance and heritability estimates were affected by both (Met + Cys):Lys ratio and strain. We observed that a G × E interaction was present, with changes in the breeding value ranking. Therefore, genetic evaluation for carcass traits should be performed under the same (Met + Cys):Lys ratio in which quails are raised., (© 2016 Blackwell Verlag GmbH.)

Our objective was to evaluate carcass and meat characteristics of Guzerat-crossbred bulls finished in feedlot. Carcasses from 18 bulls, randomly selected from a larger group of 36 bulls, consisting of F1 Guzerat×Holstein ("Guzholstein"); F1 Guzerat×Nellore ("Guzonell"); and 1/2 Simmental+1/4 Guzerat+1/4 Nellore (Three-Cross; n=6 each group) were used. Cold carcass weight was greater (P=0.01) for Three-Cross compared with "Guzonell" and "Guzholstein". Three-Cross carcasses had greater (P<0.01) rib-eye-area and 100-kg-adjusted rib-eye-area among groups. Longissimus lumborum length did not differ (P>0.05) among groups, but depth was greater (P<0.01) for Three-Cross compared with other groups. "Guzholtein" had lesser (P=0.05) shear force compared with "Guzonell", with Three-Cross being intermediate. We conclude that "Guzholstein" is an adequate option for producers willing to finish this kind of genetic group, as it is comparable or better than Bos indicus crosses and B. indicus×Bos taurus bulls., (Copyright © 2015 Elsevier Ltd. All rights reserved.)

YopH tyrosine phosphatase, a virulence factor produced by pathogenic species of Yersinia, is an attractive drug target. In this work, three oxidovanadium(IV) complexes were assayed against recombinant YopH and showed strong inhibition of the enzyme in the nanomolar range. Molecular modeling indicated that their binding is reinforced by H-bond, cation-π, and π-π interactions conferring specificity toward YopH. These complexes are thus interesting lead molecules for phosphatase inhibitor drug discovery.

Our objective was to study the effect of different roughage sources produced in a tropical environment on intake, digestibility, and ruminal parameters of crossbred bulls. Five rumen-fistulated 30-mo-old Holstein × Bos indicus bulls (average BW =459 ± 32.5 kg) were utilized in a 5 × 5 randomized Latin square design. The experiment consisted of five 19-d experimental periods (10 d for adaptation and 9 d for data collection) and 5 treatments. Experimental diets consisted of corn (Zea mays L.) silage (CS), Brachiaria grass (Brachiaria decumbens Stapf.) silage (BGS), elephant grass (Pennisetum purpureum Schumach.) silage(EGS), Tifton 85 (Cynodon spp.) hay (T85), and fresh sugarcane (Saccharum officinarum L.; SC). Diets were formulated to have approximately 11% CP (DM basis)using a mixture of urea and ammonium sulfate (9:1 ratio) that was diluted in water and then mixed with roughage. Intake, ruminal outflow, digestibility coefficients,ruminal pool, intake, passage (kp) and digestion rates, microbial parameters, and pH data were assessed.D ry matter intake was greater (P < 0.01) for CS compared with SC. Among treatments, lesser (P < 0.01)potentially digestible NDF, ash- and protein-free NDF(apNDF), and digestible ash- and protein-free NDF intake values were observed for SC. Ruminal outflows of DM and nonfibrous carbohydrates were lesser (P <0.01) for SC compared with other treatments. Dry matter apparent digestibility and ruminal digestibility did not differ (P > 0.27) among treatments. No differences (P = 0.11) were observed for ruminal apNDF pool and ruminal kp of apNDF (P = 0.06) among treatments. No treatment effect (P > 0.28) was observed for DM and nitrogen of rumen-isolated bacteria, and microbial efficiency was greater (P = 0.02) for BGS among treatments.A treatment × time interaction (P < 0.01) was found for ruminal pH, with a quadratic pattern in terms of time for CS, BGS, EGS, and T85, whereas pH values for CA linearly decreased as a function of time. In summary, these roughages, when supplemented with an additional nitrogen sources, show similar overall apparent digestibility and ruminal pool values, but conversely, bulls fed sugarcane had less overall nutrient intake and lower ruminal outflow and digestion rate values. We conclude that fresh-cut sugarcane, when provided with urea, can affect digestion rate and, consequently, impact nutrient intake.

Biological nitrogen fixation from the legume-rhizobia symbiosis is one of the main sources of fixed nitrogen on land environments. Diazotrophic bacteria taxonomy has been substantially modified by the joint use of phenotypic, physiological and molecular aspects. Among these molecular tools, sequencing and genotyping of genomic regions such as 16S rDNA and repetitive conserved DNA regions have boosted the accuracy of species identification. This research is a phylogenetic study of diazotrophic bacteria from sabiá (Mimosa caesalpiniifolia Benth.), inoculated with soils from five municipalities of the Brazilian Northeast. After bacterial isolation and morphophysiological characterization, genotyping was performed using REP, ERIC and BOX oligonucleotides and 16S rDNA sequencing for genetic diversity identification. A 1.5b Kb fragment of the 16S rDNA was amplified from each isolate. Morphophysiological characterization of the 47 isolates created a dendrogram, where isolate PE-GR02 formed a monophyletic branch. The fingerprinting conducted with BOX, ERIC and REP shows distinct patterns, and their compilation created a dendrogram with diverse groups and, after blasting in GenBank, resulted in genetic identities ranging from 77 to 99 % with Burkholderia strains. The 16S rDNA phylogenetic tree constructed with these isolates and GenBank deposits of strains recommended for inoculant production confirm these isolates are distinct from the previously deposited strains, whereas isolates PE-CR02, PE-CR4, PE-CR07, PE-CR09 and PE-GE06 were the most distinct within the group. Morphophysiological characterization and BOX, ERIC and REP compilation enhanced the discrimination of the isolates, and the 16S rDNA sequences compared with GenBank confirmed the preference of Mimosa for Burkholderia diazotrophic bacteria.

Three experiments were conducted to examine the effects of injectable trace minerals (ITM) on measures of trace mineral status and performance in pre- and postweaned Brangus-crossbred beef calves. In Exp. 1, calves were assigned to treatments in alternating birth order (n = 150; 75/treatment), consisting of a 1-mL subcutaneous injection of ITM (MultiMin 90; MultiMin USA, Inc., Fort Collins, CO) or sterile saline. The ITM formulation consisted of 60, 10, 15, and 5 mg/mL of Zn, Mn, Cu, and Se. Treatments were readministered at 100 and 200 d of age. Calf BW was recorded at birth and on d 100, 150, 200, and 250 (weaning). Trace mineral status was assessed in liver biopsy samples (n = 12/treatment) collected on d 150, 200, and 250. Administration of ITM had no impact on BW gain (P ≥ 0.55) but did result in greater (P ≤ 0.02) concentrations of liver Cu and Se and lesser (P = 0.05) liver Fe concentrations compared to saline-injected calves. In Exp. 2, 24 heifers were selected from the weaned calves of Exp. 1 (n = 12/treatment) and transported 1,600 km. Remaining on their original treatments, heifers were administered 5 mL of ITM or saline following transport (d 0). Blood samples, for acute phase protein (APP) analysis, were collected on d 0, 1, 3, 6, 9, and 13 and liver biopsy samples for assessment of trace mineral status on d 13. Plasma APP concentrations increased in all calves following weaning and transport but concentrations were greatest (P < 0.05) in ITM- vs. saline-injected heifers on d 6 and 9. Liver concentrations of Cu, Se, and Zn were greater (P ≤ 0.04) but ADG lesser (P = 0.05) for heifers receiving ITM vs. saline. In Exp. 3, 34 heifers, without previous exposure to ITM, were enrolled in a 177-d development study (n = 17/treatment). Treatments consisted of 2.5-mL injections of ITM or sterile saline on d 0, 51, and 127. Humoral immune response to an injection of porcine red blood cells (PRBC) was evaluated on d 51. Trace mineral status was evaluated in liver biopsy samples collected on d 177. Overall heifer ADG, PRBC antibody titers, and liver Se concentrations were greatest (P ≤ 0.06) for ITM vs. control heifers. Collectively, these studies demonstrate an increased trace mineral status, a greater humoral response to novel antigen, and a heightened APP response to weaning and transport stress in pre- and postweaned beef calves administered ITM.

Our objective was to evaluate production, nutritive value and carrying capacity of piatã grass (Brachiaria brizantha cv. BRS Piatã), and performance of Nellore heifers in agrosilvopastoral systems (ASPS) with three eucalyptus (Eucalyptus urograndis) tree densities, during winter, spring, summer, and fall. Three integrated systems were evaluated: ASPS-1 (357 trees ha(-1)), ASPS-2 (227 trees ha(-1)), and CON (5 trees ha(-1)). In each system, two sward heights were evaluated: short and tall. A total of 80 11-month-old Nellore heifers were randomly allocated in a randomized split-plot block, 3 × 2 factorial. Greater dry matter availability was observed on CON pastures during the fall season. Greater percentage of leaf lamina was detected on ASPS-1 with short sward height and greater during summer, compared with other seasons. A greater forage production was observed between tree rows and for tall sward height. Spring was the season with less forage nutritive value. Average daily gain was greater during summer and fall. Gain per hectare and stocking rate were greater on CON system and on ASPS-2. Pastures with short sward height had greater gain per hectare and stocking rate. Agrosilvopastoral systems with intermediate tree density seem to be a good choice for producers willing to diversify their revenue sources without decreasing animal production.

YopH plays a relevant role in three pathogenic species of Yersinia. Due to its importance in the prevention of the inflammatory response of the host, this enzyme has become a valid target for the identification and development of new inhibitors. In this work, an in-house library of 283 synthetic compounds was assayed against recombinant YopH from Yersinia enterocolitica. From these, four chalcone derivatives and one sulfonamide were identified for the first time as competitive inhibitors of YopH with binding affinity in the low micromolar range. Molecular modeling investigations indicated that the new inhibitors showed similar binding modes, establishing polar and hydrophobic contacts with key residues of the YopH binding site., (Copyright © 2013 Elsevier Masson SAS. All rights reserved.)

Tuberculosis (TB) is a major infectious disease caused by Mycobacterium tuberculosis (Mtb). According to the World Health Organization (WHO), about 1.8 million people die from TB and 10 million new cases are recorded each year. Recently, a new series of naphthylchalcones has been identified as inhibitors of Mtb protein tyrosine phosphatases (PTPs). In this work, 100 chalcones were designed, synthesized, and investigated for their inhibitory properties against MtbPtps. Structure-activity relationships (SAR) were developed, leading to the discovery of new potent inhibitors with IC(50) values in the low-micromolar range. Kinetic studies revealed competitive inhibition and high selectivity toward the Mtb enzymes. Molecular modeling investigations were carried out with the aim of revealing the most relevant structural requirements underlying the binding affinity and selectivity of this series of inhibitors as potential anti-TB drugs.

Tuberculosis (TB) is a major cause of morbidity and mortality throughout the world, and it is estimated that one-third of the world's population is infected with Mycobacterium tuberculosis. Among a series of tested compounds, we have recently identified five synthetic chalcones which inhibit the activity of M. tuberculosis protein tyrosine phosphatase A (PtpA), an enzyme associated with M. tuberculosis infectivity. Kinetic studies demonstrated that these compounds are reversible competitive inhibitors. In this work we also carried out the analysis of the molecular recognition of these inhibitors on their macromolecular target, PtpA, through molecular modeling. We observed that the predominant determinants responsible for the inhibitory activity of the chalcones are the positions of the two methoxyl groups at the A-ring, that establish hydrogen bonds with the amino acid residues Arg17, His49, and Thr12 in the active site of PtpA, and the substitution of the phenyl ring for a 2-naphthyl group as B-ring, that undergoes pi stacking hydrophobic interaction with the Trp48 residue from PtpA. Interestingly, reduction of mycobacterial survival in human macrophages upon inhibitor treatment suggests their potential use as novel therapeutics. The biological activity, synthetic versatility, and low cost are clear advantages of this new class of potential tuberculostatic agents.

Objective: To report the first case of disseminated Trichosporon spp infection in a newborn infant in Brazil, discussing a few aspects concerning management and treatment. A new spectrum of pathogens associated with severe infections in neonatal ICU has arisen, afflicting mainly newborn infants weighing less than 1,000 g at birth. Infection with Trichosporon asahii is rare and often fatal in this group of patients., Description: A case of Trichosporon spp fatal infection in a newborn weighing 815 g at birth is reported. Literature search in the main databases returned only nine articles, reporting 14 cases of infection with this fungus in preterm newborns., Conclusions: The rate of invasive fungal infection is around 6% in this group of patients, Trichosporon infection being a likely occurrence. Mortality rate in these cases is extremely high, but early treatment with triazole antifungals improves prognosis significantly.

Refractory hypotension is frequent in very low-birth weight infants, whose hypothalamic-pituitary-adrenal axis has been suggested to be immature. The objective of the present study was to evaluate basal cortisol and 17-alpha-OH-progesterone in the first 36 h of life in preterm infants with and without refractory hypotension (mean arterial blood pressure below the lower limit for gestational age throughout the study despite aggressive volume expansion and use of vasopressors). Thirty-five infants with