José Luiz de Lima Filho

Nanoencapsulation of quercetin and resveratrol into elastic liposomes.

Based on the fact that quercetin (QUE) and resveratrol (RES) induce a synergic inhibition of the adipogenesis and increase apoptosis in adipocytes, and that sodium deoxycholate (SDC) has necrotic effects, the nanoencapsulation of QUE and RES into SDC-elastic liposomes is proposed as a new approach for dissolving the subcutaneous fat. The concentration of constituents and the effect of the drug incorporation into cyclodextrin inclusion complexes on the stability of QUE/RES-loaded liposomes were studied. The best liposomal formulation reduced the use of phosphatidylcholine and cholesterol in 17.7% and 68.4%, respectively. Liposomes presented a mean diameter of 149nm with a polydispersion index of 0.3. The zeta potential of liposomes was slightly negative (-13.3mV) due to the presence of SDC in the phospholipid bilayer. Encapsulation efficiency of QUE and RES into liposomes was almost 97%. To summarize, QUE/RES-loaded elastic liposomes are stable and suitable for subcutaneous injection, thereby providing a new strategy for reducing subcutaneous fat.

DEFB1 gene polymorphisms and increased risk of HIV-1 infection in Brazilian children

In our study we analysed three single nucleotide polymorphisms (SNPs) in the 5′ untranslated region (UTR) of the DEFB1 gene, namely −52(G/A) −44(C/G) and −20(G/A), in three groups of northeastern Brazilian children in order to assess their role in HIV-1 infection. Our results allowed us to hypothesize that the SNPs located in the 5′ UTR of the DEFB1 gene can be employed as a marker of risk for HIV-1 infection.

Flow-injection amperometric determination of dopamine in pharmaceuticals using a polyphenol oxidase biosensor obtained from soursop pulp

The amperometric determination of dopamine (Do) in pharmaceuticals formulations by flow injection analysis (FIA) is proposed. An enzymatically modified carbon paste electrode constituted by 25% (w/w) of polyphenol oxidase obtained from Annona muricata L. tissue, 30% (w/w) of graphite, 30% (w/w) of silicone and 15% (w/w) of 7,7,8,8 tetracyanoquinodimethane (TCNQ), was used as flow-through detector. The flow amperometric detection was carried out at a potential of 0.10 V (vs. Ag/AgCl) when an injected sample volume of 250 microl was inserted on a 0.3 M phosphate buffer carrier solution (pH 7.8) flowing at 2.5 ml/min. The developed biosensor showed good stability and reproducibility, enabling up to 500 determinations in 60 days, without considerable loss of enzymatic activity. The FIA system presented a linear response to Do concentrations in the interval from 2 x 10(-2) to 2 x 10(-4) M, with relative standard deviations lower than 1.5%. The kinetic parameter K(M) for the soluble and immobilized enzyme was 1.45 x 10(-2) and 1.91 x 10(-2) M, respectively. In the analyses of different commercially pharmaceutical formulations a relative deviation lower than about 3.4% was obtained.

Immunological's host profile for HPV and Chlamydia trachomatis, a cervical cancer cofactor.

Over 100 different genotypes of human papillomavirus (HPV) have been isolated to date, while Chlamydia trachomatis is the most common bacterial sexually-transmitted pathogen. This review considers evidence that C. trachomatis infection became a cofactor for HPV establishment and the development of cervical intraepithelial neoplasia.

Shrimp Waste Extract and Astaxanthin: Rat Alveolar Macrophage, Oxidative Stress and Inflammation

UNLABELLED Astaxanthin is a carotenoid known to have antioxidant and antiinflammatory properties. This study examined if shrimp astaxanthin modulates the production of superoxide (O(-)(2)), nitric oxide (NO), and tumor necrosis factor-α (TNF-α) in rat alveolar macrophages. The oxidative effect was induced by phorbol myristate acetate and lipopolysacharide. The treatment was compared with superoxide dismutase, butylated hydroxytoluene, commercial astaxanthin, N-nitric-L-arginine methyl ester and L- canavanine, all administered as a 43.5-μg/mL dose in the presence of 1% EtOH/0.5% DMSO. All treatments maintained cell viability, as observed in the MTT assay, and shrimp extract increased the viable alveolar macrophages to 168%. Shrimp extract and commercial astaxanthin showed a suppressive effect on the generation of both free radicals O(-)(2) and NO, while purified shrimp astaxanthin was specific to NO. TNF-α secretion was correlated with NO production. However, in this correlation, the shrimp extract completely inhibited TNF-α. In the light of these findings, the antioxidant action demonstrated in this study suggests that the shrimp extract could be considered as a promising source of bioactive substances with antioxidant and anti-inflammatory activity. PRACTICAL APPLICATION The hydrolysis process of shrimp waste generates bioactive products that add economic value to shrimp processing, mainly because they may have applications in nutraceutical and animal feed industry.

Salmonella spp., importante agente patogênico veiculado em alimentos

A ocorrencia de doencas transmitidas por alimentos (DTAs) tem sido foco de discussoes nos ultimos anos, devido a preocupacao mundial com estrategias que permitam seu controle e, consequentemente, garantam a colocacao de produtos seguros no mercado consumidor. A Salmonella spp. e um dos microrganismos mais amplamente distribuidos na natureza, sendo o homem e os animais seus principais reservatorios naturais, com ocorrencia de sorotipos regionais, reconhecidos como salmoneloses, e considerado como um dos principais agentes envolvidos em surtos de origem alimentar em paises desenvolvidos. O aumento da incidencia da salmonelose provocada por alimentos contaminados demonstra que, na atualidade, apesar dos avancos tecnologicos alcancados, este problema ainda ocorre mundialmente. As aves e bovinos sao responsaveis pela maior disseminacao desse agente patogenico. A ampla distribuicao da Salmonella spp. entre os animais, a existencia de portadores assintomaticos e sua permanencia no ambiente e nos alimentos contribuem para que este microrganismo assuma um papel de grande relevância na saude publica mundial e, portanto, programas permanentes de controle e erradicacao devem ser adotados.The occurrence of foodborne diseases has been a matter of discussion over the last years due to the worldwide concern with strategies for their control and for ensuring that safe food products reach the consumer. Salmonella spp. is among the most widespread microorganisms in nature, having man and animals as main natural reservoirs. With occurrence of regional serotypes causing salmonellosis, this pathogen is considered one of the main agents responsible for outbreaks of foodborne disease in the developing countries. The increasing incidence of salmonellosis caused by contaminated food has shown that, despite the recent technological improvements, this problem still occurs in all countries. Cattle and poultry are the main responsible for the transmission of this pathogenic agent. Due to its wide distribution in animals, the existence of asymptomatic carriers and its presence in foodstuff and in the environment Salmonella spp. represents a significant public health problem worldwide calling for permanent control programs and eradication strategies.

IL-18 gene promoter polymorphism is involved in HIV-1 infection in a Brazilian pediatric population

In our study, we identified a polymorphism (C-607A) in the promoter region of the IL-18 gene that shows different frequencies between human immunodeficiency virus (HIV)-1-infected children and healthy controls in a pediatric Brazilian population. The presence of the −607 C allele correlates to HIV-1 infection and confers an increased risk of infection in subjects carrying the single nucleotide polymorphism.

Selective Cathepsin S Inhibition Attenuates Atherosclerosis in Apolipoprotein E–Deficient Mice with Chronic Renal Disease

Chronic renal disease (CRD) accelerates the development of atherosclerosis. The potent protease cathepsin S cleaves elastin and generates bioactive elastin peptides, thus promoting vascular inflammation and calcification. We hypothesized that selective cathepsin S inhibition attenuates atherogenesis in hypercholesterolemic mice with CRD. CRD was induced by 5/6 nephrectomy in high-fat high-cholesterol fed apolipoprotein E-deficient mice. CRD mice received a diet admixed with 6.6 or 60 mg/kg of the potent and selective cathepsin S inhibitor RO5444101 or a control diet. CRD mice had significantly higher plasma levels of osteopontin, osteocalcin, and osteoprotegerin (204%, 148%, and 55%, respectively; P < 0.05), which were inhibited by RO5444101 (60%, 40%, and 36%, respectively; P < 0.05). Near-infrared fluorescence molecular imaging revealed a significant reduction in cathepsin activity in treated mice. RO5444101 decreased osteogenic activity. Histologic assessment in atherosclerotic plaque demonstrated that RO5444101 reduced immunoreactive cathepsin S (P < 0.05), elastin degradation (P = 0.01), plaque size (P = 0.01), macrophage accumulation (P < 0.01), growth differentiation factor-15 (P = 0.0001), and calcification (alkaline phosphatase activity, P < 0.01; osteocalcin, P < 0.05). Furthermore, cathepsin S inhibitor or siRNA significantly decreased expression of growth differentiation factor-15 and monocyte chemotactic protein-1 in a murine macrophage cell line and human primary macrophages. Systemic inhibition of cathepsin S attenuates the progression of atherosclerotic lesions in 5/6 nephrectomized mice, serving as a potential treatment for atherosclerosis in patients with CRD.

Association between MBL2 gene functional polymorphisms and high-risk human papillomavirus infection in Brazilian women

We studied the association between high-risk human papillomavirus (HPV) infection and MBL2 functional polymorphisms in a group of 180 high-risk HPV-infected women and 180 healthy control subjects. The most frequent high-risk HPV genotypes were 16 (47.2%), 31 (11.7%), 33 (5%), and 18 (2.2%), respectively. Of the 180 HPV-infected women, 99 presented with uterine cervical cancer and 81 did not. No differences in MBL2 genotype or in allelic or haplotype frequencies were found between HPV patients who developed cervical uterine cancer and those who did not. When considering combined genotypes grouped according to MBL production (designated as high, low, and deficient producers), we detected a significant difference between healthy controls and high-risk HPV-positive patients, the latter group showing increased frequencies of deficient-producer genotypes (14.4% vs 9.4% in the healthy control group, corrected p = 0.04). In conclusion, a correlation between MBL2 polymorphisms and high-risk HPV infection was found in this study.

Kinetic and Thermodynamic Investigation on Ascorbate Oxidase Activity and Stability of a Cucurbita maxima Extract

The kinetic and thermodynamic properties of ascorbate oxidase (AO) activity and stability of a Cucurbita maxima extract were investigated. Activity tests performed at 25 °C using initial ascorbic acid concentration in the range 50–750 μM allowed estimating the Michaelis constant for this substrate (Km = 126 μM) and the maximum initial rate of ascorbic acid oxidation (A0,max = 1.57 mM min−1). The main thermodynamic parameters of the enzyme reaction (ΔH* = 10.3 kJ mol−1; ΔG* = 87.2 kJ mol−1; ΔS* = –258 J mol−1 K−1) were estimated through activity tests performed at 25–48 °C. Within such a temperature range, no decrease in the initial reaction rate was detected. The long‐term thermostability of the raw extract was then investigated by means of residual activity tests carried out at 10–70 °C, which allowed estimating the thermodynamic parameters of the irreversible enzyme inactivation as well (ΔH*D = 51.7 kJ mol−1; ΔG*D = 103 kJ mol−1; ΔS*D = –160 J mol−1 K−1). Taking into account the specific rate of AO inactivation determined at different temperatures, we also estimated the enzyme half‐life (1047 min at 10 °C and 21.2 min at 70 °C) and predicted the integral activity of a continuous system using this enzyme preparation. This work should be considered as a preliminary attempt to characterize the AO activity of a C. maxima extract before its concentration by liquid‐liquid extraction techniques.