Estudo Farmacognóstico e Screening Biológico de Solanum lycocarpum St. Hill (Solanaceae)

Pós-graduação em Ciências Farmacêuticas - FCFAR

Mucopolysaccharidoses in northern Brazil: targeted mutation screening and urinary glycosaminoglycan excretion in patients undergoing enzyme replacement therapy

Mucopolysaccharidoses (MPS) are rare lysosomal disorders caused by the deficiency of specific lysosomal enzymes responsible for glycosaminoglycan (GAG) degradation. Enzyme Replacement Therapy (ERT) has been shown to reduce accumulation and urinary excretion of GAG, and to improve some of the patients' clinical signs. We studied biochemical and molecular characteristics of nine MPS patients (two MPS I, four MPS II and three MPS VI) undergoing ERT in northern Brazil. The responsiveness of ERT was evaluated through urinary GAG excretion measurements. Patients were screened for eight common MPS mutations, using PCR, restriction enzyme tests and direct sequencing. Two MPS I patients had the previously reported mutation p.P533R. In the MPS II patients, mutation analysis identified the mutation p.R468W, and in the MPS VI patients, polymorphisms p.V358M and p.V376M were also found. After 48 weeks of ERT, biochemical analysis showed a significantly decreased total urinary GAG excretion in patients with MPS I (p < 0.01) and MPS VI (p < 0.01). Our findings demonstrate the effect of ERT on urinary GAG excretion and suggest the adoption of a screening strategy for genotyping MPS patients living far from the main reference centers.

Desenvolvimento de métodos quantitativos e de sistemas de screening para a determinação de glifosato

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

A simple and rapid screening method for sulfonamides in honey using a flow injection system coupled to a liquid waveguide capillary cell

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Identification of Novel Compounds Inhibiting Chikungunya Virus-Induced Cell Death by High Throughput Screening of a Kinase Inhibitor Library

Chikungunya virus (CHIKV) is a mosquito-borne arthrogenic alphavirus that causes acute febrile illness in humans accompanied by joint pains and in many cases, persistent arthralgia lasting weeks to years. The re-emergence of CHIKV has resulted in numerous outbreaks in the eastern hemisphere, and threatens to expand in the foreseeable future. Unfortunately, no effective treatment is currently available. The present study reports the use of resazurin in a cell-based high-throughput assay, and an image-based high-content assay to identify and characterize inhibitors of CHIKV-infection in vitro. CHIKV is a highly cytopathic virus that rapidly kills infected cells. Thus, cell viability of HuH-7 cells infected with CHIKV in the presence of compounds was determined by measuring metabolic reduction of resazurin to identify inhibitors of CHIKV-associated cell death. A kinase inhibitor library of 4,000 compounds was screened against CHIKV infection of HuH-7 cells using the resazurin reduction assay, and the cell toxicity was also measured in non-infected cells. Seventy-two compounds showing >= 50% inhibition property against CHIKV at 10 mu M were selected as primary hits. Four compounds having a benzofuran core scaffold (CND0335, CND0364, CND0366 and CND0415), one pyrrolopyridine (CND0545) and one thiazol-carboxamide (CND3514) inhibited CHIKV-associated cell death in a dose-dependent manner, with EC50 values between 2.2 mu M and 7.1 mu M. Based on image analysis, these 6 hit compounds did not inhibit CHIKV replication in the host cell. However, CHIKV-infected cells manifested less prominent apoptotic blebs typical of CHIKV cytopathic effect compared with the control infection. Moreover, treatment with these compounds reduced viral titers in the medium of CHIKV-infected cells by up to 100-fold. In conclusion, this cell-based high-throughput screening assay using resazurin, combined with the image-based high content assay approach identified compounds against CHIKV having a novel antiviral activity -inhibition of virus-induced CPE - likely by targeting kinases involved in apoptosis.

Evaluation of a simple alkaline pretreatment for screening of sugarcane hybrids according to their in vitro digestibility

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

Screening of Extracts of Leaves and Stems of Psychotria spp. (Rubiaceae) against Sitophilus zeamais (Coleoptera: Curculionidae) and Spodoptera frugiperda (Lepidoptera: Noctuidae) for Maize Protection

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Screening of wild type Streptomyces isolates able to overproduce clavulanic acid

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

Screening and determination of sulphonamide residues in bovine milk samples using a flow injection system

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Systematic Screening of Plant Extracts from the Brazilian Pantanal with Antimicrobial Activity against Bacteria with Cariogenic Relevance

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

Genomic screening of testicular germ cell tumors from monozygotic twins

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

Screening of Conditions for the Preparation of Poly( -Caprolactone) Nanocapsules Containing the Local Anesthetic Articaine

The local anesthetic articaine (ATC) is widely used in dentistry; however, its side effects can include paresthesia and nerve injury. Polymeric nanocapsules (PN) can be used as carriers for drugs, and help to reduce undesirable symptoms. The objective of this study was to evaluate the influence of different factors on the average size, polydispersion, and encapsulation efficiency of PN containing ATC. Poly(ε-caprolactone) (PCL) nanocapsules containing ATC were prepared by the oil-in-water emulsion/solvent evaporation method. The final ATC concentration was 2%. The preparation conditions were optimized using a central composite blocked cube-star design to investigate the influence of two variables at five levels, with 22 factorial points (–1 and +1), two replicates of the central point, 2×2 axial points (–1.414 and +1.414), and an orthogonal distribution, resulting in 10 experiments. The factors varied were the PVA concentration and the sonication time. The nanocapsules showed a satisfactory size range, a polydispersivity index less than 0.2, and high encapsulation efficiency. The values of the factors had no significant influence on either average size or polydispersion, although the encapsulation efficiency was significantly influenced by the sonication time. Improved formulations were identified using the central composite design, which revealed that the main consideration in selecting a suitable formulation was the encapsulation efficiency. Two of the formulations showed both high encapsulation efficiency and colloidal characteristics appropriate for the route of administration.

Screening test for antibiotics in medicinal plants (STAMP): using powdered plant materials instead of extracts

Coordenação de Aprefeiçoamento de Pessoal de Nível Superior (CAPES)