In vitro detection of hepatitis C virus in platelets from uninfected individuals exposed to the virus

Introduction ,,,,,Despite hepatocytes being the target cells of hepatitis C virus (HCV), viral ribonucleic acid RNA has been detected in other cells, including platelets, which have been described as carriers of the virus in the circulation of infected patients. Platelets do not express cluster differentiation 81 CD81, the main receptor for the virus in hepatocytes, although this receptor protein has been found in megakaryocytes. Still, it is not clear if HCV interacts with platelets directly or if this interaction is a consequence of its association with megakaryocytes. The aim of this study was to evaluate the interaction of HCV with platelets from non-infected individuals, after in vitro exposure to the virus. ,,,, ,,,, ,,,,,Methods ,,,,,Platelets obtained from 50 blood donors not infected by HCV were incubated in vitro at 37°C for 48h with serum containing 100,000IU∕mL of genotype 1 HCV. After incubation, RNA extracted from the platelets was assayed for the presence of HCV by reverse transcription – polymerase chain reaction RT-PCR. ,,,, ,,,, ,,,,,Results ,,,,,After incubation in the presence of virus, all samples of platelets showed HCV RNA. ,,,, ,,,, ,,,,,Conclusions ,,,,,The results demonstrate that, in vitro, the virus interacts with platelets despite the absence of the receptor CD81, suggesting that other molecules could be involved in this association.

Correlation between the mobility of spin-labeled peptide chains and resin solvation: An approach to optimize the synthesis of aggregating sequences

Resin solvation properties affect the efficiency of the coupling reactions in solid-phase peptide synthesis. Here we report a novel approach to evaluate resin solvation properties, making use of spin label electron paramagnetic resonance (EPR) spectroscopy. The aggregating VVLGAAIV and ING sequences were assembled in benzhydrylamine-resin with different amino group contents (up to 2.6 mmol/g) to examine the extent of chain association within the beads. These model peptidyl-resins were first labeled at their N-terminus with the amino acid spin label 2,2,6,6-tetramethylpiperidine-N-oxyl-4-amino-4-carboxylic acid (Toac). Their solvation properties in different solvents were estimated, either by bead swelling measurement or by assessing the dynamics of their polymeric matrixes through the analysis of Toac EPR spectra, and were correlated with the yield of the acylation reaction. In most cases the coupling rate was found to depend on bead swelling. Comparatively, the EPR approach was more effective. Line shape analysis allowed the detection of more than one peptide chain population, which influenced the reaction. The results demonstrated the unique potential of EPR spectroscopy not only for improving the yield of peptide synthesis, even in challenging conditions, but also for other relevant polymer-supported methodologies in chemistry and biology.

Structural and functional characterization of myotoxin I, a Lys49 phospholipase A(2) homologue from Bothrops moojeni (Caissaca) snake venom

Myotoxin-I (MjTX-I) was purified to homogeneity from the venom of Bothrops moojeni by ion-exchange chromatography on CM-Sepharose. Its molecular weight, estimated by SDS-PAGE, was 13,400 (reduced) or 26,000 (unreduced). The extinction coefficient (E-1.0 cm(1.0 mg/ml)) of MjTX-I was 1.145 at lambda = 278 nm, pH 7.0, and its isoelectric point was 8.2 at ionic strength mu = 0.1. When lyophilized and stored at 4 degrees C, dimeric, trimeric, and pentameric forms of the protein were identified by SDS-PAGE. This heterogeneous sample could be separated into three fractions by gel filtration on Sephadex 6-50. The fractions were analyzed by isoelectric focusing, immunoelectrophoresis, and amino acid composition, which indicated that heterogeneity was the result of different levels of self-association. Protein sequencing indicated that MjTX-I is a Lys49 myotoxin and consists of 121 amino acids (M-r = 13,669), containing a high proportion of basic and hydrophobic residues. It shares a high degree of sequence identity with other Lys49 PLA(2)-like myotoxins, but shows a significantly lower identity with catalytically active Asp49 PLA(2)s. The three-dimensional structure of MjTX-I was modeled based on the crystal structures of three highly homologous Lys49 PLA(2)-like myotoxins. This model showed that the amino acid substitutions are conservative, and mainly the beta-wing region, and the C-terminal extended random coil. MjTX-I displays local myotoxic and edema-inducing activities in mice, and is lethal by intraperitoneal injection, with an LD50 value of 8.5 +/- 0.8 mg/kg, In addition, it is cytotoxic to myoblasts/ myotubes in culture, and disrupts negatively charged liposomes. In comparison with the freshly prepared dimeric sample, the more aggregated forms showed significantly reduced myotoxic activity. However, the edema-inducing activity of MjTX-I was independent of molecular association. Phospholipase A(2) activity on egg yolk, as well as anticoagulant activity, were undetectable both in the native and in the more associated forms. His, Tyr, and Trp residues of the toxin were chemically modified by specific reagents. Although the myotoxic and lethal activities of the modified toxins were reduced by these treatments, neither its edema-inducing or Liposome-disrupting activities were significantly altered. Rabbit antibodies to native MjTX-I cross-reacted with the chemically modified forms, and both the native and modified MjTX-I preparations were recognized by antibodies against the C-terminal region 115-129 of myotoxin II from B. asper, a highly Lys49 PLA(2)-homologue with high sequencial similarity. (C) 2000 Academic Press.

WATER BALANCE and SPATIAL DISTRIBUTION of AN ANURAN COMMUNITY FROM BRAZIL

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Hyolith-dominated shell beds from the Lampazar Formation in NW Argentina: patterns and processes of origin in the Late Cambrian (Furongian) seas of western Gondwana.

Late Cambrian (Furongian) shell beds in the Salta Province of NW Argentina are unique because of the presence of abundant hyolith skeletal remains within them. Hyolith shell beds are located in the mid-upper part of the Lampazar Formation at the Angosto de La Quesera locality, and are the first recorded accumulations of this type in the lower Palaeozoic of the South American Andean Basin. The shell beds are of the order of several mm thick, and are laterally persistent within outcrop scale, with a few metres of lateral development. Two types of hyolith shell beds are recognised: Type 1 is a storm-dominated, event concentration, represented by dispersed to densely packed accumulations of well preserved hyolith and gastropod shells (Strepsodiscus austrinus). Hyolith conchs are current oriented with the long axes parallel to unidirectional flow on the sandstones surfaces. Type 2 shell beds are background, composite concentrations, of poorly preserved, comminuted debris of hyolith shells with associated gastropod and trilobite sclerites (dominated by Parabolina, Beltella and Leiostegium). The genesis of both shell beds was controlled primarily by physical processes, such as storms and current and/or wave agitation. The thickness, simple internal fabric and geometry shown by both accumulations are typical of Cambrian-style shell-beds.

ATTENDANCE and CO-OCCURRENCE of BIRDS FOLLOWING ARMY ANTS IN THE ATLANTIC RAIN FOREST

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

THE IMAGINARY ABOUT THE SCHOOL and THE POSSIBILITIES of AN EQUALITARIAN SCHOOL: A STUDY on THE BRAZILIAN STATE SCHOOLS

The main objective of the article is to reflect on the imaginary that the teachers of two public schools have of their students and the school they work in and the ideal of an inclusive and democratic school. In this article, we reflect on the image that the teachers of two public schools in the city of Rio Claro, São Paulo State, Brazil, have of their students and their school as an institution. We use the studies of imaginary sociology as a theoretic reference base. Castoriadis (1986); Taylor (2006); Legros et al (2007) and Baczo (1984) are the referenced authors of this article. (C) 2012 Published by Elsevier Ltd. Selection and/or peer-review under responsibility of The Association of Science, Education and Technology

Inferences about the coefficient of correlation in the standard bivariate normal distribution

The study of the association between two random variables that have a joint normal distribution is of interest in applied statistics; for example, in statistical genetics. This article, targeted to applied statisticians, addresses inferences about the coefficient of correlation (ρ) in the bivariate normal and standard bivariate normal distributions using likelihood, frequentist, and Baycsian perspectives. Some results are surprising. For instance, the maximum likelihood estimator and the posterior distribution of ρ in the standard bivariate normal distribution do not follow directly from results for a general bivariate normal distribution. An example employing bootstrap and rejection sampling procedures is used to illustrate some of the peculiarities.

Use of aspartate aminotransferase in diagnosing periodontal disease: a comparative study of clinical and microbiological parameters

The objective of this study was to assess the association between the levels of enzyme aspartate aminotransferase (AST) in gingival crevicular fluid (GCF) with the BANA hydrolysis microbiological test (Perioscan) and clinical periodontal diagnostic measurements, such as bleeding on probing, plaque index, gingival index, probing depth, and attachment level in patients with chronic periodontitis using an enzymatic test (PocketWatch). One hundred and forty-seven sites were evaluated in 22 patients with a probing depth of > or = 5 mm at selected sites. AST and BANA enzymatic tests were carried out, and clinical parameters recorded. Pearson's chi-square and Fisher's exact tests were used for statistical analysis. There was no statistical correlation between AST levels and any of the analyzed parameters. The lack of any association between the factors studied does not indicate, however, that the latter cannot be used in diagnosing the actual periodontal condition of patients and/or sites. However, more research should be carried out to evaluate the true relationship between AST and periodontal disease.

On the specific filtration mechanism of a mesoporous silica membrane, prepared with non-connecting parallel pores

We report the singular filtration properties of an ultrafiltration membrane made with mesoporous silica that exhibits cylindrical pores aligned mostly normal to the support. This membrane supported on tubular commercial macroporous alumina supports was prepared by the interfacial growth mechanism between stable silica-surfactant hybrid micelles made of the association of silica oligomers with polyethyleneoxide-based (PEO) surfactants and sodium fluoride, a well-known silica condensation catalyst [Boissière et al., An ultrafiltration membrane made with mesoporous MSU-X silica, Chem. Mater. 15 (2003) 460-463]. It appears that the combined effect of the silica nature of the membrane, whose surface charge can be easily adjusted by changing the pH and the non-connected cylindrical shape of the pores provides a new behavior in the retention properties, as proved by the filtration of polyoxyethylene polymers (PEO) with different molecular weights. Depending on the filtration conditions, a rejection rate of 80% and a steep cut-off at 2000 Da can be obtained or, on the reverse, polymers three times bigger than the pore diameter can diffuse through the membrane. This new filtration mechanism, which opens up new modes of separation modes, is explained in the light of both topology of the porous network and pH-dependent interactions between PEO polymers and silica porous media. © 2004 Elsevier B.V. All rights reserved.

Transmission analysis of candidate genes for nonsyndromic oral clefts in Brazilian parent-child triads with recurrence

Cleft lip and/or palate (CL/P) is a major congenital defect with complex etiology, including multiple genetic and environmental factors. Approximately two thirds of the cases are not accompanied by other anomalies and are called nonsyndromic (NS). In the present study, we performed transmission distortion analysis of the MSX1-CA, TGFB3-CA and MTHFR-C677T polymorphisms in 60 parent-child triads, in which the NS-CL/ P affected child had at least one affected parent. No association with genes MSX1 or TGFB3 was found, but the results were suggestive of an association of the MTHFR-C677T polymorphism with NS-CL/P. © 2006 Sociedade Brasileira de Genética.

Attributable fraction of work accidents related to occupational noise exposure in a Southeastern city of Brazil

Noise is the most frequent type of occupational exposure and can lead to both auditory and extra-auditory dysfunction as well as increasing the risk of work accidents. The purpose of this study was to estimate the attributable fraction of work accidents related to occupational noise exposure in a medium-sized city in Southeast Brazil. In this hospital-based case-control study, including 600 cases and 822 controls, the odds ratio of work accidents (controlled for several covariables) was obtained classifying occupational noise exposure into four levels and determining the prevalence at each level. Based on these data, the calculated attributable fraction was 0.3041 (95%CI: 0.2341-0.3676), i.e., 30% of work accidents in the study area were statistically associated with occupational noise exposure. The authors discuss the causes of this association and the implications for the prevention of work accidents.

Apoptosis in different gastric lesions and gastric cancer: Relationship with Helicobacter pylori, overexpression of p53 and aneuploidy

Apoptosis has an essential function in maintaining the integrity of the gastrointestinal mucosa. Its deregulation is associated with the occurrence of lesions such as in atrophic gastritis, peptic ulcers, intestinal metaplasia, and stomach tumorigenesis. Thus, the aim of the present study was to investigate the frequency of apoptotic cells (apoptotic index, AI) by using two different immunohistochemical techniques, TUNEL and anti-activated caspase-3 antibody (CPP32), in gastric dyspepsia [chronic gastritis (CG, N = 34), chronic atrophic gastritis (CAG, N = 11), gastric ulcer (GU, N = 17), and intestinal metaplasia (IM, N = 15)], normal gastric mucosae (NM, N = 8), and gastric adenocarcinoma (GC, N = 12). The relationship was investigated between the AI and Helicobacter pylori infection, diagnosed by PCR, overexpression of p53 protein determined by immunohistochemistry, and aneuploidy by fluorescence in situ hybridization, as performed by our laboratory in previous studies. No significant differences were observed in AI between the different groups, whether by the TUNEL technique (F = 1.60; P = 0.1670) or by CPP32 antibody (F = 1.70; P = 0.1420). Nonetheless, CAG and CG groups had AI statistically higher than those of normal mucosae. These two groups (CAG and CG) also showed a higher frequency of apoptosis-positive cases (TUNEL+ or CPP32+). Generally, there was no correlation between the AI detected by the TUNEL and CPP32 techniques in the groups studied, except in the GC group (r = 0.70). Moreover, there was no significant association between apoptosis and H. pylori infection, overexpression of p53 protein and aneuploidy, but the H. pylori-positive cases only of GU (P = 0.0233) and IM (P = 0.0253) groups displayed a statistically higher AI compared to H. pylori-negative NM, when the CPP32 antibody technique was used. Thus, CG and CAG have increased apoptosis, which may occur independent of an association with H. pylori infection, aneuploidy and overexpression of p53 protein. ©FUNPEC-RP.

Distribution of carabid beetles in agroecosystems across spatial scales - A review

The review focuses on the questions (1) how does the spatial heterogeneity of landscape influences carabid biodiversity, and (2) what are the main factors causing this biodiversity across nested spatial scales (study point - plant association - landscape level). The analysis of recent literature indicates that the spatial distribution of carabids differs at various spatial scales, and the factors responsible for the distribution are different. At the study point level most of the communities exhibit high variability of population density and diversity, which has no correlations with soil, and sometimes, vegetation, parameters. Most of the factors that contribute to formation of the communities are stochastic, simply because patches of a factor are much smaller than the size of a distinct carabid community. At the level of plant association, soil factors begin to play the role in driving the communities. At this level, litter depth, micro-climate and vegetation composition are the main factors. At the landscape level, geological factors, such as topography, landscape geochemistry, and history are playing important roles. As a conservation measure, spatial heterogeneity should be kept at all spatial scales at the same time to maintain carabid biodiversity in agricultural areas.

Evaluation of yield advantage indexes in carrot-lettuce intercropping systems

The aim of this study was to estimate the effect of the association between carrot and lettuce crops in several intercropping systems through different yield advantage indexes, as well as to assess which system is better for the environmental resources management with respect to productivity and economic indicators. A 'group balanced block' experimental design was used, with four replications. Cultivars of lettuce crispleaf ('Lucy Brown', 'Tainá', 'Laurel' and 'Verônica') and looseleaf ('Babá de Verão', 'Maravilha das Quatro Estações', 'Elisa' and 'Carolina') groups were evaluated in intercropping systems with 'Alvorada' and 'Brasilia' carrot cultivars. The land equivalent ratio (LER) and yield efficiency index (YEI) were estimated, besides economic indicators such as gross (GI) and net (NI) income, modified monetary advantage (MMA), return rate (RR) and profit margin (PM). The evaluated indexes showed that carrot is the dominant and lettuce the dominated crop. Higher biological/agronomic efficiency indexes and economic indicators were observed in intercropping systems with 'Brasilia' carrot as component crop and that based on the crispleaf lettuce group.