The Mode of Action of Recombinant Mycobacterium tuberculosis Shikimate Kinase: Kinetics and Thermodynamics Analyses

Tuberculosis remains as one of the main cause of mortality worldwide due to a single infectious agent, Mycobacterium tuberculosis. The aroK-encoded M. tuberculosis Shikimate Kinase (MtSK), shown to be essential for survival of bacilli, catalyzes the phosphoryl transfer from ATP to the carbon-3 hydroxyl group of shikimate (SKH), yielding shikimate-3-phosphate and ADP. Here we present purification to homogeneity, and oligomeric state determination of recombinant MtSK. Biochemical and biophysical data suggest that the chemical reaction catalyzed by monomeric MtSK follows a rapid-equilibrium random order of substrate binding, and ordered product release. Isothermal titration calorimetry (ITC) for binding of ligands to MtSK provided thermodynamic signatures of non-covalent interactions to each process. A comparison of steady-state kinetics parameters and equilibrium dissociation constant value determined by ITC showed that ATP binding does not increase the affinity of MtSK for SKH. We suggest that MtSK would more appropriately be described as an aroL-encoded type II shikimate kinase. Our manuscript also gives thermodynamic description of SKH binding to MtSK and data for the number of protons exchanged during this bimolecular interaction. The negative value for the change in constant pressure heat capacity (ΔCp) and molecular homology model building suggest a pronounced contribution of desolvation of non-polar groups upon binary complex formation. Thermodynamic parameters were deconvoluted into hydrophobic and vibrational contributions upon MtSK:SKH binary complex formation. Data for the number of protons exchanged during this bimolecular interaction are interpreted in light of a structural model to try to propose the likely amino acid side chains that are the proton donors to bulk solvent following MtSK:SKH complex formation. © 2013 Rosado et al.

Evaluation of metals in water and sediments of micro-basins in the city of Americana, São Paulo state, Brazil

Water pollution found in major rivers in Brazil has its origin from urban sewage discharges and industrial effluent, carried out by small streams and rivers crossing cities. Therefore, studies related to hydrographic micro-basins offer the opportunity to establish environmental management strategies for restoring water resources, based on diagnosis of the water quality. Despite this understanding, few studies in urban and rural areas have been performed in a systematic manner in Brazilian micro-basins. The main goal of this research was to diagnose the water resources in micro-basins in the region of the district of Americana, São Paulo state, Brazil, through the quantification of metals in water and sediment. The methodology was based on the investigation of metals (Ba, Cd, Cr, Cu, Fe, Mn, Ni, Pb, Zn), in four micro-basins, determined by inductively coupled plasma optical emission spectrometry (ICP-OES). The most significant result showed high concentration levels of chromium (969 μg L-1), downstream of the discharge of sewage from the city of Nova Odessa. This concentration in the river was above the allowed limit of Brazilian regulation agency (50 μg L-1 for Cr). Also high levels of Cr were found in the sediment (98.9 μg g-1) collected at the same monitored site. These results are important indicators of environmental performance and anthropogenic activities to help the government establish environmental management strategies aimed at the reduction of water pollution. © 2013 WIT Press.

Nanostructured materials based on mesoporous silica and mesoporous silica/apatite as osteogenic growth peptide carriers

The aim of this work was the preparation of inorganic mesoporous materials from silica, calcium phosphate and a nonionic surfactant and to evaluate the incorporation and release of different concentrations of osteogenic growth peptide (OGP) for application in bone regeneration. The adsorption and release of the labeled peptide with 5,6-carboxyfluorescein (OGP-CF) from the mesoporous matrix was monitored by fluorescence spectroscopy. The specific surface area was 880 and 484 m2 g- 1 for pure silica (SiO) and silica/apatite (SiCaP), respectively; the area influenced the percentage of incorporation of the peptide. The release of OGP-CF from the materials in simulated body fluid (SBF) was dependent on the composition of the particles, the amount of incorporated peptide and the degradation of the material. The release of 50% of the peptide content occurred at around 4 and 30 h for SiCaP and SiO, respectively. In conclusion, the materials based on SiO and SiCaP showed in vitro bioactivity and degradation; thus, these materials should be considered as alternative biomaterials for bone regeneration. © 2013 Elsevier B.V.

In vitro and in vivo evaluation of the activity of pineapple (Ananas comosus) on Haemonchus contortus in Santa Inês sheep

The development of resistance to anthelmintics has prompted research into alternative methods of controlling intestinal nematodes in ruminants. This study aimed to assess the activity of Ananas comosus on Haemonchus contortus in Santa Inês sheep. The aqueous extract of pineapple skin (AEPS), bromelain from pineapple stems (B4882) and residue from pineapple processing was evaluated in in vitro and in vivo tests. The enzymatic activity of substances was analyzed by the azocasein method. The egg hatch test (EHT) and larval development test (LDT) were performed using the Embrapa2010 isolate of H. contortus. In the in vivo test, 36 sheep artificially infected with H. contortus were divided into six groups: G1: 2g/kg BW of the aqueous extract administered for three days; G2: 2g/kg BW of the industrial pineapple residue for 60 days; G3: 180mg/animal of bromelain in a single dose; G4: negative control I; G5: positive control (levamisole phosphate); and G6: negative control II. The eggs per gram (EPG) in the feces were counted till 28 days after treatment. LC50 and LC90 were obtained by the probit procedure, while the in vivo test results were analyzed by GLM. The aqueous extract in the in vitro and in vivo test, the bromelain and industrial residue presented 0.102, 0.157, 1.864 and 0.048 enzyme units/mL, respectively. In the egg hatch test, the LC50 and LC90 were respectively 31 and 81mg/mL for the aqueous extract and 0.50 and 2mg/mL for bromelain. In the larval development test, the LC50 and LC90 were respectively 1.7 and 7.3mg/mL for the aqueous extract and 0.019 and 0.086mg/mL for bromelain. In the in vivo test, the general efficacies of the treatments in relation to the negative control were 22.6%, 42.2%, 3.65% and 89% for the aqueous extract, industrial pineapple residue, bromelain and positive control respectively. The transformed EPG values were 3.19±0.59, 3.32±0.25, 2.85±0.66, 3.44±0.50, 2.28±0.93 and 2.75±0.94 for the aqueous extract, industrial residue, bromelain, negative control I, positive control and negative control II respectively. The results for all the treated groups differed significantly (p<0.05) from the positive control, and although the residue presented efficacy of 42.2%, there was no statistical difference (p>0.05) in relation to the negative control. Therefore, both the aqueous extract and bromelain were effective in vitro, but showed reduced anthelmintic efficacy in vivo. For the pineapple residue, the 42.2% in vivo efficacy in reducing the EPG and the possibility of reducing environmental contamination through reuse of industrial residue indicate it can also be useful for control of this parasite. © 2013 Elsevier B.V.

Avaliação da taxa de remoção de material dissolvido devido ao intemperismo químico da formação Serra Geral na bacia do Ribeirão Preto

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Estudo da degradação dos solos em áreas de pastagens no município de Porto Velho (RO)

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Thermal and structural study of glasses in the binary system TeO2-Pb(PO3)(2)

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Bacterial Cellulose/Chondroitin Sulfate for Dental Materials Scaffolds

Bacterial cellulose (BC) has become established as a remarkably versatile biomaterial and can be used in a wide variety of scientific applications, especially for medical devices. In this work, the bacterial cellulose fermentation process is modified by the addition of chondroitin sulfate (1% w/w) to the culture medium before the bacteria are inoculated. Besides, biomimetic precipitation of calcium phosphate of biological interest from simulated body fluid on bacterial cellulose was studied. Chondroitin sulfate influences in bacterial cellulose were analyzed using transmission infrared spectroscopy (FTIR), XRD (X-ray diffraction) and scanning electron microscopy (SEM). FTIR analysis showed interaction between chondroitin sulfate, bacterial cellulose and calcium phosphate and XRD demonstrated amorphous calcium phosphate and carbonated apatite on bacterial cellulose nanocomposites. SEM images confirmed incorporation of calcium phosphate in bacterial celluloe nanocomposite surface and uniform spherical calcium phosphate particles. Future experiments with cells adhesion and viability are in course.

ANTIDIARRHEAL MECHANISM AND IONIC PROFILE OF CARPOLOBIA LUTEA ETHANOLIC STEM-BARK EXTRACT IN RATS

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Influence of Abiotic Factors and Floral Resource Availability on Daily Foraging Activity of Bees

In this study, the daily and seasonal influences of abiotic factors and the amount of floral resources on the foraging frequency of bees were determined. The experiments were performed, during 12 consecutive months, in the main floral sources used by bees in a secondary forest fragment. The foraging frequency of each bee species on flowers of each plant was recorded for 20-min periods, every hour. To verify whether the foraging activity is influenced by abiotic factors, Pearson's correlation analysis and linear regression tests were performed for the dominant bee species. Temperature and luminosity were the two main abiotic factors regulating foraging activities of bees. A positive correlation was found between the foraging frequency of most bees and these two variables. Conversely, the foraging activity was influenced neither by the relative humidity nor by the wind speed. The activity of each species depends on a combination of factors that include not only abiotic variables, but also the amount of floral resources available during the day, body size, and behavior of each visitor. After a certain period of the day, the scarcity of floral resources produced by most plants can stimulate the bees to forage in the flowers early in subsequent days, which may occur before the period in which the abiotic conditions are really favorable.

Crystallization and preliminary X-ray diffraction analysis of a novel sphingomyelinase D from Loxosceles gaucho venom

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Identification and enzymatic characterization of acid phosphatase from Burkholderia gladioli

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

The stigma and prejudice of leprosy: influence on the human condition

Introduction: To analyze the knowledge, feelings and perceptions involving patients affected by leprosy, as a better understanding of these factors may be useful to decrease the stigma and prejudice associated with the condition. Methods: The study cohort consisted of 94 patients who underwent treatment for leprosy at the Health Units in the City of Cuiaba, Mato Grosso (MT), Brazil. The study questionnaire included items to collect information on socio-demographic data, knowledge about the disease, stigma, prejudice, self-esteem and quality of life of leprosy patients. Bivariate analyses were used to assess the data based on the chi-square test with a 5% significance threshold. Results: The results revealed that the study population consisted predominantly of males (55.3%) with an income between 1 and 3 times the minimum wage (67%). The survey respondents reported that the most significant difficulties related to the treatment were the side effects (44.7%) and the duration of the treatment (28.7%). A total of 72.3% of the subjects were knowledgeable about the disease, of whom 26.6% had the leprosy reaction. Stigma and prejudice were cited by 93.6% of the participants. Based on the responses, 40.4% of patients reported being depressed and sad, and 69.1% of the subjects encountered problems at work after being diagnosed. A total of 45.7% of the patients rated their quality of life between bad and very bad. Conclusions: Our results suggest that leprosy causes suffering in patients beyond pain and discomfort and greatly influences social participation.

Bacterial cellulose biocomposites for guided tissue regeneration

Bacterial cellulose (BC) has become established as a remarkably versatile biomaterial and can be used in a wide variety of scientific applications, especially for medical devices. In this work, the bacterial cellulose fermentation process is modified by the addition of chondroitin sulfate and hyaluronic acid (1% w/w) to the culture medium before the bacteria is inoculated. Besides, biomimetic precipitation of calcium phosphate of biological interest from simulated body fluid on bacterial cellulose was studied. Chondroitin sulfate and hyaluronic acid influences in bacterial cellulose were analyzed using transmission infrared spectroscopy (FTIR), XRD (X-ray diffraction) and scanning electron microscopy (SEM). FTIR analysis showed interaction between bacterial cellulose nanobiocomposites and calcium phosphate and XRD demonstrated amorphous calcium phosphate and calcium chloride on bacterial cellulose nanobiocomposites. SEM images confirmed incorporation of calcium phosphate in bacterial cellulose nanobiocomposites surface with different calcium phosphate particles morphology.

In vitro and in vivo biological behavior of CDHA/OCP/β-TCP scaffold

The biological behavior of a new bioactive material composed of calcium-deficient hydroxyapatite, octacalcium phosphate, and beta-tricalcium phosphate was investigated by in vitro indirect and direct cytotoxicity, cell adhesion and proliferation tests, and by in vivo subcutaneous and bone implantation in rats. The results of the in vitro studies showed that the material is biocompatible and no cytotoxic. Slightly poorer initial cell adhesion and lower cell proliferation than in control was observed, which were attributed to the reactivity and roughness of the material surface, In vivo results showed that the material is biodegradable and bioactive in bone tissue, but only biocompatible and partially biodegradable in soft tissue.