Determining the structural basis for specificity of ligands using crystallographic screening

Crystallographic screening has been used to identify new inhibitors for potential target for drug development. Here, we describe the application of the crystallographic screening to assess the structural basis of specificity of ligands against a protein target. The method is efficient and results in detailed crystallographic information. The utility of the method is demonstrated in the study of the structural basis for specificity of ligands for human purine nucleoside phosphorylase (PNP). Purine nucleoside phosphorylase catalyzes the phosphorolysis of the N-ribosidic bonds of purine nucleosides and deoxynucleosides. This enzyme is a target for inhibitor development aiming at T-cell immune response modulation and has been submitted to extensive structure-based drug design. This methodology may help in the future development of a new generation of PNP inhibitors.

A non-invasive method for silencing gene transcription in honeybees maintained under natural conditions

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

A rapid, environmentally friendly, and reliable method for pesticide analysis in high-fat samples

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

Determination of snake-necked turtle Hydromedusa tectifera (Cope, 1870) (Testudines: Chelidae) plasma protein concentrations by refractometry and the biuret method.

The purpose of this study was to evaluate the accuracy of hand-held refractometer in determining plasma protein concentrations in South American snake-necked turtle (Hydromedusa tectifera) as compared with the standard biuret method. The results indicated that plasma protein values may be accurately determined in snake- necked turtle with a hand-held refractometer.

Tests of one Brazilian facial reconstruction method using three soft tissue depth sets and familiar assessors

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Gap size measurement: the proposal of a new field method

In the tropical Atlantic Forest, 42 canopy gaps had their areas estimated using four different field methods of measurement: Runkle, Brokaw and Green [Runkle, J.R., 1981. Gap formation in some old-growth forests of the eastern United States. Ecology 62, 1041-1051; Brokaw, N.V.L., 1982. The definition of treefall gap and its effect on measures of forest dynamics. Biotropica 14, 158-160; Green, P.T., 1996. Canopy Gaps in rain forest on Christmas Island, Indian Ocean: size distribution and methods of measurement. J. Trop. Ecol. 12, 427-434] and a new method proposed in this work. It was found that within the same gap delimitation, average gap size varied from 56.0 up to 88.3 m(3) while total sum of gap area varied from 2351.3 to 3707.9 m(3) Differences among all methods and between pairs of method proved to be statistically significant. As a consequence, gap size-class distribution was also different between methods. When one method is held as a standard, deviation on average values of gap size ranged between 11.8 and 59.7% as deviations on single gap size can reach 172.8%. Implications on forest dynamics were expressed by the forest turnover rate that was 24% faster or 15% slower depending on the method adopted for gap measurement. Based on my results and on methods' evaluation, the use of a new method is proposed here for future research involving the measure of gap size in forest ecosystems. Finally, it is concluded that forest comparisons disregarding the influence of different methods of gap measurement should be reconsidered. (c) 2005 Elsevier B.V. All rights reserved.

A simple method for estimating global DNA methylation using bisulfite PCR of repetitive DNA elements

We report a method for studying global DNA methylation based on using bisulfite treatment of DNA and simultaneous PCR of multiple DNA repetitive elements, such as Alu elements and long interspersed nucleotide elements (LINE). The PCR product, which represents a pool of approximately 15000 genomic loci, could be used for direct sequencing, selective restriction digestion or pyrosequencing, in order to quantitate DNA methylation. By restriction digestion or pyrosequencing, the assay was reproducible with a standard deviation of only 2% between assays. Using this method we found that almost two-thirds of the CpG methylation sites in Alu elements are mutated, but of the remaining methylation target sites, 87% were methylated. Due to the heavy methylation of repetitive elements, this assay was especially useful in detecting decreases in DNA methylation, and this assay was validated by examining cell lines treated with the methylation inhibitor 5-aza-2'deoxycytidine (DAC), where we found a 1-16% decrease in Alu element and 18-60% LINE methylation within 3 days of treatment. This method can be used as a surrogate marker of genome-wide methylation changes. In addition, it is less labor intensive and requires less DNA than previous methods of assessing global DNA methylation.

Growth rates in modern speleothems from Santana Cave, Brazil, by the Pb-210-method

The Santana Cave is located at the Upper Ribeira Touristic State Park (PETAR-Parque Estadual Turistico do Alto Ribeira) in southern São Paulo State, Brazil. This paper describes Pb-210 activity concentration data in soda straw stalactites samples collected at Salao das Flores in Santana Cave that is a fossil tributary of the cave river. Non-expensive alpha counting following some analytical steps for extracting and depositing Pb-210 were used for providing the Pb-210 data. In the analyzed samples, Pb-210 values of increasingly older samples fitted an exponential curve, thus suggesting that the production of Pb-210 has been constant with time. Also, the near-ideal fit indicated that the growth was uniform and there was no break in the continuous growth. The soda straw growth rates were determined from the best fit to the exponential curve through the Pb-210 activity concentration. The results of the measurements allowed estimate a longitudinal rate corresponding to 1.3 mm/yr and a lateral rate of 0.01 mm/yr, which permitted calculate times of 70 years and 317-498 years for their formation, respectively. The lateral growth rate is compatible with values from studies of chemical weathering rates held under laboratory and natural conditions. (C) 2011 Elsevier Ltd. All rights reserved.

An alternative genotyping method using dye-labeled universal primer to reduce unspecific amplifications

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

Determination of ring-necked pheasant (Phasianus colchicus) serum protein concentrations by refractometry and the biuret method

The purpose of this study was to evaluate the accuracy of hand-held refractometer in determining serum protein concentrations in ring-necked pheasants (Phasianus colchicus) as compared with the standard biuret method. The results indicated that serum protein values may be accurately determined in ring-necked pheasants with a hand-held refractometer. © Asian Network for Scientific Information, 2008.

Corpo e psicossexualidade: metáforas da cultura

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

TiO2/CuO films obtained by citrate precursor method for photocatalytic application

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