The cytotoxicity of methacryloxylethyl cetyl ammonium chloride, a cationic antibacterial monomer, is related to oxidative stress and the intrinsic mitochondrial apoptotic pathway

Antibacterial monomers incorporated in dentin bonding systems may have toxic effects on the pulp. Thus, the cytotoxicity of antibacterial monomers and its underlying mechanisms must be elucidated to improve the safety of antibacterial monomer application. The influence of an antibacterial monomer, methacryloxylethyl cetyl ammonium chloride (DMAE-CB), on the vitality of L929 mouse fibroblasts was tested using MTT assay. Cell cycle progression was studied using flow cytometry. Production of intracellular reactive oxygen species (ROS) after DMAE-CB treatment was measured using 2,7-dichlorodihydrofluorescein diacetate staining and flow cytometry analysis. Loss of mitochondrial membrane potential, disturbance of Bcl-2 and Bax expression, as well as release of cytochrome C were also measured using flow cytometry analysis or Western blot to explore the possible involvement of the mitochondrial-related apoptotic pathway. DMAE-CB elicited cell death in a dose-dependent manner and more than 50% of cells were killed after treatment with 30 µM of the monomer. Both necrosis and apoptosis were observed. DMAE-CB also induced G1- and G2-phase arrest. Increased levels of intracellular ROS were observed after 1 h and this overproduction was further enhanced by 6-h treatment with the monomer. DMAE-CB may cause apoptosis by disturbing the expression of Bcl-2 and Bax, reducing the mitochondrial potential and inducing release of cytochrome C. Taken together, these findings suggest that the toxicity of the antibacterial monomer DMAE-CB is associated with ROS production, mitochondrial dysfunction, cell cycle disturbance, and cell apoptosis/necrosis.

Increased immunohistochemical expression of YKL-40 in the spleen of patients with portal hypertension

YKL-40 has been identified as a growth factor in connective tissue cells and also a migration factor in vascular smooth muscle cells. To a large extent, the increase of serum YKL-40 is attributed to liver fibrosis and asthma. However, the relationship of the expression and clinical/prognostic significance of YKL-40 to the splenomegaly of patients with portal hypertension is unclear. In the present study, the expression of YKL-40 was studied by immunohistochemistry in 48 splenomegaly tissue samples from patients with portal hypertension and in 14 normal spleen specimens. All specimens were quickly stored at -80°C after resection. Primary antibodies YKL-40 (1:150 dilution, rabbit polyclonal IgG) and MMP-9 (1:200 dilution, rabbit monoclonal IgG) and antirabbit immunoglobulins (HRP K4010) were used in this study. The relationship of clinicopathologic features with YKL-40 is presented. The expression of YKL-40 indicated by increased immunochemical reactivity was significantly up-regulated in splenomegaly tissues compared to normal spleen tissues. Overexpression of YKL-40 was found in 68.8% of splenomegaly tissues and was significantly associated with Child-Pugh classification (P = 0.000), free portal pressure (correlation coefficient = 0.499, P < 0.01) and spleen fibrosis (correlation coefficient = 0.857, P < 0.01). Further study showed a significant correlation between YKL-40 and MMP-9 (correlation coefficient = -0.839, P < 0.01), indicating that YKL-40 might be an accelerator of spleen tissue remodeling by inhibiting the expression of MMP-9. In conclusion, YKL-40 is an important factor involved in the remodeling of spleen tissue of portal hypertension patients and can be used as a therapeutic target for splenomegaly.

Analysis of acylcarnitine profiles in umbilical cord blood and during the early neonatal period by electrospray ionization tandem mass spectrometry

Acylcarnitine profiling by electrospray ionization tandem mass spectrometry (ESI-MS/MS) is a potent tool for the diagnosis and screening of fatty acid oxidation and organic acid disorders. Few studies have analyzed free carnitine and acylcarnitines in dried blood spots (DBS) of umbilical cord blood (CB) and the postnatal changes in the concentrations of these analytes. We have investigated these metabolites in healthy exclusively breastfed neonates and examined possible effects of birth weight and gestational age. DBS of CB were collected from 162 adequate for gestational age neonates. Paired DBS of heel-prick blood were collected 4-8 days after birth from 106 of these neonates, the majority exclusively breastfed. Methanol extracts of DBS with deuterium-labeled internal standards were derivatized before analysis by ESI-MS/MS. Most of the analytes were measured using a full-scan method. The levels of the major long-chain acylcarnitines, palmitoylcarnitine, stearoylcarnitine, and oleoylcarnitine, increased by 27, 12, and 109%, respectively, in the first week of life. Free carnitine and acetylcarnitine had a modest increase: 8 and 11%, respectively. Propionylcarnitine presented a different behavior, decreasing 9% during the period. The correlations between birth weight or gestational age and the concentrations of the analytes in DBS were weak (r £ 0.20) or nonsignificant. Adaptation to breast milk as the sole source of nutrients can explain the increase of these metabolites along the early neonatal period. Acylcarnitine profiling in CB should have a role in the early detection of metabolic disorders in high-risk neonates.

Obtenção de nova fonte de peroxidase de folha de Copaifera langsdorffii Desf. com alta atividade

Objetivou-se neste trabalho extrair peroxidase de folha de Copaifera langsdorffii (COP), medir sua atividade, compará-la com a peroxidase de raiz forte (Horseradish peroxidase - HRP) e determinar o pH ótimo, a melhor solução extratora e o efeito de aditivos sobre a atividade da COP. Os resultados mostraram que a COP atingiu 81,6% da atividade de HRP e a faixa de pH ótimo foi de 5,5 a 6,0. A melhor solução extratora da enzima foi o tampão fosfato de sódio 50 mM, pH 6,0 e o melhor aditivo foi o PVPP. Concluindo, a COP apresenta atividade mais alta que outras peroxidases de diferentes fontes citadas na literatura.

Biosensor for determination of glucose in real samples of beverages

A biosensor was developed for spectrophotometric determination of glucose concentrations in real samples of orange juice energetic drinks, and sport drinks. The biosensor consisted of glucose oxidase (GOD) and horseradish peroxidase (HRP) immobilized onto polyaniline activated with glutaraldehyde (PANIG). Immobilization parameters were optimized for GOD, and maximum immobilization yield was 16% when 5.0 mg of PANIG and 8.9 U prepared in 0.1 mol.L-1 sodium phosphate buffer (pH 7.0) reacted for 60 minutes at 4 °C with gentle stirring. The linear operational range for glucose determination using optimized operational parameters was between 0.05 and 6.0 mg.mL-1 with a very good reproducibility of response. The results obtained in the biosensor were compared with those obtained using free enzymes (commercial kits) and then validated through statistical analysis using the Tukey test (95% confidence interval).

Development and chemical and sensory characterization of pumpkin seed flour-based cereal bars

Pumpkin (Cucurbita maxima), popularly known as squash, is a widely grown vegetable in Brazil. In this study, pumpkin seed flours (PSF) with different granulometries were used: PSF 1 (medium granulometry) and PSF 2 (coarse granulometry) in the preparation of cereal bars (CB) with different combinations with brown oats. Five formulations were prepared: CB-1 (control - 25% brown oats and 0% PSF); CB-2 (12.5% PSF 1 and 12.5% brown oats); CB-3 (25% PSF 1 and 0% brown oats); CB-4 (12.5% PSF 2 and 12.5% brown oats); and CB-5 (25% PSF 2 and 0% brown oats). The acceptance test results were analyzed in a conventional preference mapping which indicated that the bars CB-2 and CB-5 received mostly the maximum hedonistic score. With the objective of developing a cereal bar replacing oats with PSF, the bars CB-2 and CB-5 were compared to the conventional bar CB-1. The cereal bars CB-2 and CB-5 showed an increase in crude protein (87.5% and 62.5%) and in dietary fiber (77% and 44%), respectively. These results allowed the classification of CB-2 and CB-5 as fiber sources; they can, therefore, be classified as light products according to the Brazilian legislation.

Desnutrição na insuficiência renal crônica: qual o melhor método diagnóstico na prática clínica?

INTRODUÇÃO: A desnutrição protéico-calórica, o processo inflamatório sistêmico e os distúrbios metabólicos são frequentes em pacientes com insuficiência renal crônica em terapia dialítica, contribuindo para sua morbimortalidade. MATERIAL E MÉTODOS: No presente estudo, a prevalência de desnutrição em pacientes renais crônicos em hemodiálise em um único centro no Nordeste do Brasil foi avaliada segundo três diferentes técnicas de avaliação subjetiva global (ASG), o índice de massa corporal (IMC), o percentual de peso atual em relação ao ideal, a adequação ao percentil 50 da prega cutânea tricipital (PCT), da circunferência do braço (CB), da circunferência muscular do braço (CMB), a albumina pré-diálise, o ângulo de fase e o percentual de massa celular corporal (MCC). A correlação do diagnóstico nutricional realizado através da ASG com as medidas antropométricas, bioquímicas e bioimpedância elétrica foi pesquisada. RESULTADOS: Foram avaliados 58 pacientes, sendo 30 do sexo feminino (51,7%), com idade média de 49 anos. A prevalência de desnutrição segundo os diferentes métodos variou entre 12,1% a 94,8%. A ASG clássica teve uma concordância moderada no diagnóstico nutricional com a ASG gerada pelo paciente, IMC com ponto de corte em 22,0 kg/m² e CMB; regular com o IMC com ponto de corte em 18,5 kg/m², adequação do peso atual em relação ao ideal, CB e ângulo de fase; e ruim com a ASG adaptada ao renal, PCT e percentual de MCC. CONCLUSÕES: Os métodos de avaliação nutricional comumente utilizados na prática clínica têm restrições na população em diálise, tendo em vista os diferentes percentuais obtidos com os diferentes métodos. Estudos longitudinais, prospectivos, pesquisando a associação dos marcadores nutricionais com eventos adversos como hospitalização e mortalidade, devem continuar sendo realizados para maior esclarecimento do problema.

Punção da fístula arteriovenosa com a técnica em casa de botão com agulha romba

INTRODUÇÃO: A canulação da fístula arteriovenosa (FAV) pode ser realizada pelas técnicas de punção por área, rotatória ou em casa de botão (CB); entretanto, a técnica ideal ainda não foi estabelecida. OBJETIVO: Avaliar as dificuldades e complicações na introdução da técnica de punção em CB em FAV construída com veia nativa. MÉTODOS: 16 pacientes com idade de 57±14 anos, em hemodiálise há 63±38 meses foram submetidos à punção em CB. Na fase de formação do túnel (T), as punções foram feitas com agulha cortante (AC) e na fase de manutenção com agulha romba (AR). Nas duas fases, os pacientes foram avaliados para a intensidade da dor em uma escala de 0 a 10. RESULTADOS: O nº de sessões de HD para formação do T foi de 9,5 ± 1,5 e o número de sessões na fase de manutenção foi de 29,7 ± 0,8. Nas 152 HD para formação do T não ocorreram complicações significativas. Durante 475 HD com AR as complicações foram: resistência na punção (7,6%), punção com AC por opção do puncionador (5,7%), troca de AR para AC durante a punção (4,2%) e sangramento local durante a HD (0,8%). Um paciente necessitou antibioticoterapia. A mediana do índice de dor foi 4 na fase de formação do T e 2 na fase de manutenção. Os valores de Kt/V pré- e pós-alteração na técnica de punção não foram diferentes (1,48 ± 0,27 e 1,45 ± 0,23). CONCLUSÃO: A implantação da punção em CB com AR é tecnicamente fácil, apresenta poucas complicações, reduz a dor e não induz variação na dose de diálise.