Study of the interaction between hydroxymethylnitrofurazone and 2-hydroxypropyl-beta-cyclodextrin

Chagas disease is a serious health problem in Latin America. Hidroxymethylnitrofurazone (NFOH) is a nitrofurazone prodrug more active than nitrofurazone against Trypanosoma cruzi. However, NFOH presents low aqueous solubility, high photodecomposition and high toxicity. The present work is focused on the characterization of an inclusion complex of NFOH in 2-hydroxypropyl-beta-cyclodextrin (HP-beta-CD). The complexation with HP-beta-CD was investigated using reversed-phase liquid chromatography, solubility isotherms and nuclear magnetic resonance. The retention behavior was analyzed on a reversed-phase C-18 column, using acetonitrile-water (20/80, v/v) as the mobile phase, in which HP-beta-CD was incorporated as a mobile phase additive. The decrease in the retention times with increasing concentrations of HP-beta-CD enables the determination of the apparent stability constant of the complex (K = 6.2 +/- 0.3 M-1) by HPLC. The solubility isotherm was studied and the value for the apparent stability constant (K = 7.9 +/- 0.2 M-1) was calculated. The application of continuous variation method indicated the presence of a complex with 1:1 NFOH:HP-beta-CD stoichiometry. The photostability study showed that the formation of an inclusion complex had a destabilizing effect on the photodecomposition of NFOH when compared to that of the "free" molecule in solution. The mobility investigation (by NMR longitudinal relaxation time) gives information about the complexation of NFOH with HP-beta-CD. In preliminary toxicity studies, cell viability tests revealed that inclusion complexes were able to decrease the toxic effect (p < 0.01) caused by NFOH. (c) 2008 Elsevier B.V. All rights reserved.

Determination of neutral polymorphisms (frameworks) of the human beta globin gene in beta thalassaemias by PCR/DGGE

Purpose: Considering the importance of type beta thalassaemias as hereditary syndromes of high significance in different populations of Mediterranean origin and, by extension, in the Brazilian population, the objective of the present study was to determine by PCR/DGGE the gene structures responsible for neutral polymorphisms (frameworks) observed in the human beta globin gene associated with the mutations responsible for type beta thalassaemias in a sample of the Brazilian population and, more specifically, of the population of the State of São Paulo. Patients and methods: Thirty individuals with beta thalassaemic mutations were analyzed: 22 mutations were in codon 39 (C->T), 5 in IVS1-110 (G->A), 2 in IVS1-6 (T->C) and 1 in IVS1-1 (G->A). DNA was extracted and selective amplification was performed by PCR extending from position IVS1 nt 46 to IVS2 nt 126 (474 pb). The product was then analyzed by polyacrylamide gel electrophoresis on a denaturing 10-60% urea/formamide gradient. Results: The results demonstrated that, as expected, the mutations responsible for type beta thalassaemia observed in this population are of Mediterranean origin, with 73% distribution represented by codon 39,17% by IVS1-110, 7% by IVS1-6 and 3% by IVS1-1. In turn, framework distribution seems to indicate a higher frequency of Fr 1-1 in codon 39 and IVS1-110, of Fr 1-3 in IVS1-6 and of Fr 1-2 in IVS1-1. Conclusions: These results permit us to conclude that gene amplification by PCR followed by DGGE is an appropriate method for the separation of DNA molecules that differ even by a single base change and therefore can be utilized to detect the alterations observed in the human beta globin gene. This methodology shows that, using only a pair of primers, it is possible to define the frameworks that are observed in the beta globin gene.

Naphthopyranone glycosides from Paepalanthus bromelioides

Two new naphtho[2,3-C]pyran-1-one glycosides, paepalantine-9-O-β-D- glucopyranoside and paepalantine-9-O-β-D-allopyranosyl(1 → 6)glucopyranoside, were isolated from an ethanolic extract of capitula from Paepalanthus bromelioides and identified from their spectrometric data.

Purification and characterization of two β-glucosidases from the thermophilic fungus Thermoascus aurantiacus

β-Glucosidase from the fungus Thermoascus aurantiacus grown on semi-solid fermentation medium (using ground corncob as substrate) was partially purified in 5 steps-ultrafiltration, ethanol precipitation, gel filtration and 2 anion exchange chromatography runs, and characterized. After the first anion exchange chromatography, β-glucosidase activity was eluted in 3 peaks (Gl-1, Gl-2, Gl-3). Only the Gl-2 and Gl-3 fractions were adsorbed on the gel matrix. Gl-2 and Gl-3 exhibited optimum pH at 4.5 and 4.0, respectively. The temperature optimum of both glucosidases was at 75-80°C. The pH stability of Gl-2 (4.0-9.0) was higher than Gl-3 (5.5-8.5); both enzyme activities showed similar patterns of thermostability. Under conditions of denaturing gel chromatography the molar mass of Gl-2 and Gl-3 was 175 and 157 kDa, respectively. Using 4-nitrophenyl β-D-glucopyranoside as substrate, Km values of 1.17 ± 0.35 and 1.38 ± 0.86 mmol/L were determined for Gl-2 and Gl-3, respectively. Both enzymes were inhibited by Ag+ and stimulated by Ca2+.

p38 MAPK regulates IL-1β induced IL-6 expression through mRNA stability in osteoblasts

Osteoblast-derived IL-6 functions in coupled bone turnover by supporting osteoclastogenesis favoring bone resorption instead of bone deposition. Gene regulation of IL-6 is complex occurring both at transcription and post-transcription levels. The focus of this paper is at the level of mRNA stability, which is important in IL-6 gene regulation. Using the MC3T3-E1 as an osteoblastic model, IL-6 secretion was dose dependently decreased by SB203580, a p38 MAPK inhibitor. Steady state IL-6 mRNA was decreased with SB203580 (2 μM) ca. 85% when stimulated by IL-1β (1-5 ng/ ml). These effects require de novo protein synthesis as they were inhibited by cycloheximide. p38 MAPK had minor effects on proximal IL-6 promoter activity in reporter gene assays. A more significant effect on IL-6 mRNA stability was observed in the presence of SB203580. Western blot analysis confirmed that SB203580 inhibited p38 MAP kinase, in response to IL-1β in a dose dependent manner in MC3T3-E1 cells. Stably transfected MC3T3-E1 reporter cell lines (MC6) containing green fluorescent protein (GFP) with the 3′untranslated region of IL-6 were constructed. Results indicated that IL-1β, TNFα, LPS but not parathyroid hormone (PTH) could increase GFP expression of these reporter cell lines. Endogenous IL-6 and reporter gene eGFP-IL-6 3′UTR mRNA was regulated by p38 in MC6 cells. In addition, transient transfection of IL-6 3′UTR reporter cells with immediate upstream MAP kinase kinase-3 and -6 increased GFP expression compared to mock transfected controls. These results indicate that p38 MAPK regulates IL-1β-stimulated IL-6 at a post transcriptional mechanism and one of the primary targets of IL-6 gene regulation is the 3′UTR of IL-6.

Polimorfismo da beta-lactoglobulina não afeta as caracterí sticas físico-químicas e a estabilidade do leite bovino

The objective of this study was to evaluate the effect of beta-lactoglobulin polymorphism, breed and seasonality on physico-chemical characteristics and stability of bovine milk. Six Holstein-Zebu (H-Z) and five Holstein dairy herds were selected, of which 660 Holstein and 293 crossbred Holstein-Zebu milk and blood samples were collected, adding up 953 samples, distributed in two collecting periods at rainy and two at dry seasons. Each milk sample was analyzed for titratable acidity, pH, freezing point and milk ethanol stability, at the following ethanol concentrations: 70, 76, 80 and 84GL. Individual blood samples were submitted to polymerase chain reaction (PCR) for the determination of beta-lactoglobulin polymorphism. No effect of beta-lactoglobulin polymorphism was observed in physico-chemical characteristics of milk, in both studied breed. Breed effect for Holstein and H-Z, respectively, was observed on titratable acidity (16.16 and 17.07°D, while effect of seasonality (for rainy and dry seasons, respectively) was also observed on freezing point (-0.5411 and -0.5376°H). Effects of breed and seasonality on milk stability were observed (Holstein-Zebu milk was less stable on dry season), however, no effect of beta-lactoglobulin on milk stability was observed.

Beta-carotene stability during drying and storage of cassava and sweet potato

The effect of blanching on the β-carotene stability during drying and storage of cassava and sweet potato was evaluated. The orange-fleshed sweet potato showed good retention of β-carotene during the blanching and drying (100% and 96%, respectively), but lower retention (84% and 91%) was observed in cassava. Cassava also showed lower β-carotene stability than sweet potato during the storage of unblanched dried samples. β-Carotene content of dried cassava was reduced from 8.6 μg/g to traces in 20 days of storage while the initial amount of dried sweet potato (463 μg/g) was reduced by about 45% (210 μg/g). Blanching did not affect the β-carotene retention during the drying, but enhanced the stability of this carotenoid during the storage of dried samples at room temperature, especially in cassava. The initial levels of blanched-dried cassava and sweet potato (7.8 and 513 μg/g, respectively) took 70 days to fall by around 50%.

VEGF-C expression in oral cancer by neurotransmitter-induced activation of beta-adrenergic receptors

The aim of this study was to investigate the expression of vascular endothelial growth factor type C (VEGF-C) in oral squamous cell carcinoma (OSCC) cell lines through norepinephrine-induced activation of beta-adrenergic receptors. Human OSCC cell lines (SCC-9 and SCC-25) expressing beta-adrenergic receptors were stimulated with different concentrations of norepinephrine (0.1, 1, and 10 μM) and 1 μM of propranolol, and analyzed after 1, 6, and 24 h. VEGF-C gene expression and VEGF-C production in the cell supernatant were evaluated by real-time PCR and by ELISA, respectively. The results showed that beta-adrenergic receptor stimulation by different concentrations of norepinephrine or blocking by propranolol did not markedly alter VEGF-C expression by SCC-9 and SCC-25 cells. VEGF-C protein levels produced by oral malignant cell lines after stimulation with different norepinephrine concentrations or blocking with propranolol was statistically similar (p > 0.05) to those of the control group (nonstimulated OSCC cell lines). Our findings suggest that stimulation of beta-adrenergic receptors by means of norepinephrine does not seem to modulate the VEGF-C expression in OSCC cell lines. These findings reinforce the need for further studies in order to understand the responsiveness of oral cancer to beta-adrenergic receptor stimulation or blockage, especially with regard to VEGF-C production. © 2012 International Society of Oncology and BioMarkers (ISOBM).

Efeito da proteína Beta-Conglicinina da soja no metabolismo lipídico de animais submetidos à dieta hipercolesterolêmica

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Expressão fenotípica da homozigose para hemoglobina S em relação aos haplótipos da beta globina, polimorfismos da glutationa S-transferase e enzimas de detoxificação

Pós-graduação em Genética - IBILCE

Expressão da trealose-6-fosfato sintase (TPS) em cana-de-açúcar (Saccharum spp) sob estresse hídrico

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Análise de crescimento, curva de absorção de macronutrientes (N, P e K) e teor de beta- ecdisona em fáfia (Pfaffia glomerata (Spreng). Pedersen em função de adubação orgânica

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Suplementação alimentar com 'beta'-glucano e mananoligossacarídeo para tilápias do Nilo em tanques-rede

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

Produção de beta-glucanases por Trichoderma harzianum Rifai para obtenção gluco-oligossacarídeos a partir de botriosferana

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

Comportamento mecânica da mola T de Beta-titânio: influência da marca comercial e do alivio de tensão estrutural

Pós-graduação em Ciências Odontológicas - FOAR