Protection of plasmid DNA by a Ginkgo biloba extract from the effects of stannous chloride and the action on the labeling of blood elements with technetium-99m

Ginkgo biloba extract (EGb) is a phytotherapeutic agent used for the treatment of ischemic and neurological disorders. Because the action of this important extract is not fully known, assays using different biological systems need to be performed. Red blood cells (RBC) are labeled with technetium-99m (Tc-99m) and used in nuclear medicine. The labeling depends on a reducing agent, usually stannous chloride (SnCl2). We assessed the effect of different concentrations of EGb on the labeling of blood constituents with Tc-99m, as sodium pertechnetate (3.7 MBq), and on the mobility of a plasmid DNA treated with SnCl2 (1.2 µg/ml) at room temperature. Blood was incubated with EGb before the addition of SnCl2 and Tc-99m. Plasma (P) and RBC were separated and precipitated with trichloroacetic acid, and soluble (SF-P and SF-RBC) and insoluble (IF-P and IF-RBC) fractions were isolated. The plasmid was incubated with Egb, SnCl2 or EGb plus SnCl2 and agarose gel electrophoresis was performed. The gel was stained with ethidium bromide and the DNA bands were visualized by fluorescence in an ultraviolet transilluminator system. EGb decreased the labeling of RBC, IF-P and IF-RBC. The supercoiled form of the plasmid was modified by treatment with SnCl2 and protected by 40 mg/ml EGb. The effect of EGb on the tested systems may be due to its chelating action with the stannous ions and/or pertechnetate or to the capability to generate reactive oxygen species that could oxidize the stannous ion.

Spectral analysis of the electromyograph of the erector spinae muscle before and after a dynamic manual load-lifting test

The aim of the present study was to assess the spectral behavior of the erector spinae muscle during isometric contractions performed before and after a dynamic manual load-lifting test carried out by the trunk in order to determine the capacity of muscle to perform this task. Nine healthy female students participated in the experiment. Their average age, height, and body mass (± SD) were 20 ± 1 years, 1.6 ± 0.03 m, and 53 ± 4 kg, respectively. The development of muscle fatigue was assessed by spectral analysis (median frequency) and root mean square with time. The test consisted of repeated bending movements from the trunk, starting from a 45º angle of flexion, with the application of approximately 15, 25 and 50% of maximum individual load, to the stand up position. The protocol used proved to be more reliable with loads exceeding 50% of the maximum for the identification of muscle fatigue by electromyography as a function of time. Most of the volunteers showed an increase in root mean square versus time on both the right (N = 7) and the left (N = 6) side, indicating a tendency to become fatigued. With respect to the changes in median frequency of the electromyographic signal, the loads used in this study had no significant effect on either the right or the left side of the erector spinae muscle at this frequency, suggesting that a higher amount and percentage of loads would produce more substantial results in the study of isotonic contractions.

Looking for the GAP effect in manual responses and the role of contextual influences in reaction time experiments

When the offset of a visual stimulus (GAP condition) precedes the onset of a target, saccadic reaction times are reduced in relation to the condition with no offset (overlap condition) - the GAP effect. However, the existence of the GAP effect for manual responses is still controversial. In two experiments using both simple (Experiment 1, N = 18) and choice key-press procedures (Experiment 2, N = 12), we looked for the GAP effect in manual responses and investigated possible contextual influences on it. Participants were asked to respond to the imperative stimulus that would occur under different experimental contexts, created by varying the array of warning-stimulus intervals (0, 300 and 1000 ms) and conditions (GAP and overlap): i) intervals and conditions were randomized throughout the experiment; ii) conditions were run in different blocks and intervals were randomized; iii) intervals were run in different blocks and conditions were randomized. Our data showed that no GAP effect was obtained for any manipulation. The predictability of stimulus occurrence produced the strongest influence on response latencies. In Experiment 1, simple manual responses were shorter when the intervals were blocked (247 ms, P < 0.001) in relation to the other two contexts (274 and 279 ms). Despite the use of choice key-press procedures, Experiment 2 produced a similar pattern of results. A discussion addressing the critical conditions to obtain the GAP effect for distinct motor responses is presented. In short, our data stress the relevance of the temporal allocation of attention for behavioral performance.

Measurement of annexin V uptake and lactadherin labeling for the quantification of apoptosis in adherent Tca8113 and ACC-2 cells

Phosphatidylserine (PS) exposure occurs during the cell death program and fluorescein-labeled lactadherin permits the detection of PS exposure earlier than annexin V in suspended cell lines. Adherent cell lines were studied for this apoptosis-associated phenomenon to determine if PS probing methods are reliable because specific membrane damage may occur during harvesting. Apoptosis was induced in the human tongue squamous carcinoma cell line (Tca8113) and the adenoid cystic carcinoma cell line (ACC-2) by arsenic trioxide. Cells were harvested with a modified procedure and labeled with lactadherin and/or annexin V. PS exposure was localized by confocal microscopy and apoptosis was quantified by flow cytometry. The detachment procedure without trypsinization did not induce cell damage. In competition binding experiments, phospholipid vesicles competed for more than 95 and 90% of lactadherin but only about 75 and 70% of annexin V binding to Tca8113 and ACC-2 cells. These data indicate that PS exposure occurs in three stages during the cell death program and that fluorescein-labeled lactadherin permitted the detection of early PS exposure. A similar pattern of PS exposure has been observed in two malignant cell lines with different adherence, suggesting that this pattern of PS exposure is common in adherent cells. Both lactadherin and annexin V could be used in adherent Tca8113 and ACC-2 cell lines when an appropriate harvesting procedure was used. Lactadherin is more sensitive than annexin V for the detection of PS exposure as the physical structure of PS in these blebs and condensed apoptotic cell surface may be more conducive to binding lactadherin than annexin V.

Manual and semi-automatic quantification of in vivo ¹H-MRS data for the classification of human primary brain tumors

In vivo proton magnetic resonance spectroscopy (¹H-MRS) is a technique capable of assessing biochemical content and pathways in normal and pathological tissue. In the brain, ¹H-MRS complements the information given by magnetic resonance images. The main goal of the present study was to assess the accuracy of ¹H-MRS for the classification of brain tumors in a pilot study comparing results obtained by manual and semi-automatic quantification of metabolites. In vivo single-voxel ¹H-MRS was performed in 24 control subjects and 26 patients with brain neoplasms that included meningiomas, high-grade neuroglial tumors and pilocytic astrocytomas. Seven metabolite groups (lactate, lipids, N-acetyl-aspartate, glutamate and glutamine group, total creatine, total choline, myo-inositol) were evaluated in all spectra by two methods: a manual one consisting of integration of manually defined peak areas, and the advanced method for accurate, robust and efficient spectral fitting (AMARES), a semi-automatic quantification method implemented in the jMRUI software. Statistical methods included discriminant analysis and the leave-one-out cross-validation method. Both manual and semi-automatic analyses detected differences in metabolite content between tumor groups and controls (P < 0.005). The classification accuracy obtained with the manual method was 75% for high-grade neuroglial tumors, 55% for meningiomas and 56% for pilocytic astrocytomas, while for the semi-automatic method it was 78, 70, and 98%, respectively. Both methods classified all control subjects correctly. The study demonstrated that ¹H-MRS accurately differentiated normal from tumoral brain tissue and confirmed the superiority of the semi-automatic quantification method.

Three days of intermittent stretching after muscle disuse alters the proteins involved in force transmission in muscle fibers in weanling rats

The aim of this study was to determine the effects of intermittent passive manual stretching on various proteins involved in force transmission in skeletal muscle. Female Wistar weanling rats were randomly assigned to 5 groups: 2 control groups containing 21- and 30-day-old rats that received neither immobilization nor stretching, and 3 test groups that received 1) passive stretching over 3 days, 2) immobilization for 7 days and then passive stretching over 3 days, or 3) immobilization for 7 days. Maximal plantar flexion in the right hind limb was imposed, and the stretching protocol of 10 repetitions of 30 s stretches was applied. The soleus muscles were harvested and processed for HE and picrosirius staining; immunohistochemical analysis of collagen types I, III, IV, desmin, and vimentin; and immunofluorescence labeling of dystrophin and CD68. The numbers of desmin- and vimentin-positive cells were significantly decreased compared with those in the control following immobilization, regardless of whether stretching was applied (P<0.05). In addition, the semi-quantitative analysis showed that collagen type I was increased and type IV was decreased in the immobilized animals, regardless of whether the stretching protocol was applied. In conclusion, the largest changes in response to stretching were observed in muscles that had been previously immobilized, and the stretching protocol applied here did not mitigate the immobilization-induced muscle changes. Muscle disuse adversely affected several proteins involved in the transmission of forces between the intracellular and extracellular compartments. Thus, the 3-day rehabilitation period tested here did not provide sufficient time for the muscles to recover from the disuse maladaptations in animals undergoing postnatal development.

Encapsulated specialty oils commercialized in São Paulo state, Brazil: evaluation of identity (fatty acid profile) and compliance of fatty acids and Vitamin E contents with nutrition labeling

Encapsulated specialty oils commercialized in São Paulo state, Brazil, were evaluated for their identity (fatty acids profile) and compliance with nutrition labeling (fatty acids and Vitamin E (alpha tocopherol) contents). Twenty one samples [flaxseed oil (6), evening primrose (5), safflower (8), borage (1), and black currant (1)] purchased from local markets or collected by the health surveillance agency were analyzed. The fatty acids and vitamin E contents were analyzed by gas chromatography with flame ionization detector and liquid chromatography with UV detector, respectively. Nine samples were adulterated (5 samples of safflower oil, 3 of flaxseed oil, and one of evening primrose). Among them, 3 flaxseed and 2 safflower oil samples were probably adulterated by the addition of soybean oil. Conjugated linoleic acid (CLA) was found in two safflower oils samples although the sale of oils with conjugated linoleic acid (CLA) is not permitted by the National Health Surveillance Agency in Brazil (ANVISA). Only two samples presented all values in compliance with nutrition labeling (one safflower oil sample and one borage oil sample). The results show that a continuous monitoring of encapsulated specialty oils commercialized in Brazil is necessary including a greater number of samples and sanitary surveillance.

Brazil nut sorting for aflatoxin prevention: a comparison between automatic and manual shelling methods

The impact of automatic and manual shelling methods during manual/visual sorting of different batches of Brazil nuts from the 2010 and 2011 harvests was evaluated in order to investigate aflatoxin prevention.The samples were tested as follows: in-shell, shell, shelled, and pieces in order to evaluate the moisture content (mc), water activity (Aw), and total aflatoxin (LOD = 0.3 µg/kg and LOQ 0.85 µg/kg) at the Brazil nut processing plant. The results of aflatoxins obtained for the manually shelled nut samples ranged from 3.0 to 60.3 µg/g and from 2.0 to 31.0 µg/g for the automatically shelled samples. All samples showed levels of mc below the limit of 15%; on the other hand, shelled samples from both harvests showed levels of Aw above the limit. There were no significant differences concerning the manual or automatic shelling results during the sorting stages. On the other hand, the visual sorting was effective in decreasing the aflatoxin contamination in both methods.

Repercussão de um protocolo fisioterapêutico intradialítico na funcionalidade pulmonar, força de preensão manual e qualidade de vida de pacientes renais crônicos

INTRODUÇÃO: A doença renal crônica consiste na perda lenta, progressiva e irreversível da função renal. É considerada um problema social e economico, pois está relacionada a inúmeras doenças associadas, bem como a altos gastos em saúde pública. Sabe-se que os pacientes dialíticos passam por longos períodos de restrição da atividade física repercutindo em disfungoes nos mais diversos sistemas e na qualidade de vida (QV). OBJETIVO: Verificar os efeitos de uma intervenção fisioterapêutica nos pacientes em hemodiálise para: função da musculatura respiratória, força de preensão manual e QV. METODOLOGIA: Estudo experimental, não randomizado, quantitativo e qualitativo; amostra de 13 pacientes, 43,69 ± 9,28 anos, submetidos à hemodiálise na Santa Casa de Diamantina/MG, selecionados por conveniência. Todos realizaram avaliação das pressões respiratórias máximas (PImáx e PEmáx) e do pico de fluxo expiratório (PFE), antes e após a fisioterapia que consistiu de três sessões semanais, durante 2 meses de: exercícios para membros superiores, com técnica de FNP e respiração diafragmática; exercícios de fortalecimento para membros inferiores e exercícios com bola exercitadora para preensão manual. O tratamento estatístico foi realizado através do teste t de Student com valor de significância em p < 0,05. RESULTADOS: As médias respectivamente das variáveis pré- e pós-intervenção foram PI , (97,69 ± 28,3 cmH2O e 98,46 ± 23,399ªdriH2O) p = 0,93; PEmáx (83,07 ± 31,19 cmH2O e 88,46 ± 14,05 cmH2O) p = 0,46 e PFE (375,38 ± 75,23 L/min e 416,15 ± 57,37 L/min) p = 0,02. A media do dinamometro pré-intervenção: 57,23 ± 17,39 kgf e pósintervenção: 56,61 ± 16,09 kgf. No SF-36, que avalia QV, observou-se melhora dos oito domínios, exceto do item 'vitalidade'. De todas as variáveis mensuradas, somente o PFE mostrou-se estatisticamente significante. CONCLUSÃO: O protocolo fisioterapêutico proposto não promoveu melhoras expressivas, do ponto de vista estatístico, nas variáveis analisadas em pacientes submetidos à hemodiálise, justificando-se em parte ao número pequeno da amostra, tempo do protocolo e intervenções propostas.

Espessura do músculo adutor do polegar como preditor da força de preensão manual nos pacientes em hemodiálise

INTODUÇÃO: A espessura do músculo adutor do polegar (EMAP) tem sido sugerida como um novo marcador de estado nutricional em diversas populações. OBJETIVO: Diante da escassez de dados sobre o uso desse marcador nos pacientes com doença renal crônica, o objetivo deste estudo foi avaliar a EMAP e sua associação com indicadores nutricionais em pacientes em hemodiálise. MÉTODOS: Foram avaliados 73 pacientes em hemodiálise (52,3 ± 17 anos, sem função renal residual). A EMAP foi aferida no braço sem o acesso vascular com o auxílio de um adipômetro. A composição corporal (bioimpedância elétrica), a força de preensão manual (dinamômetro), o estado nutricional (Avaliação Global Subjetiva) e os exames laboratoriais (creatinina, proteína total e albumina) também foram avaliados. RESULTADOS: Indivíduos com valores de EMAP acima da mediana eram em maior proporção negros/pardos, jovens e possuíam maior força de preensão manual. A EMAP correlacionou-se positivamente com a força de preensão manual, albumina sérica e massa celular (%), e negativamente com a idade. Na análise de regressão linear ajustada para sexo, idade e tempo em hemodiálise, a EMAP esteve independentemente associada com a força de preensão manual. CONCLUSÃO: A EMAP foi capaz de predizer a força de preensão manual nos pacientes em hemodiálise, o que sugere a EMAP como um marcador promissor de estado nutricional nessa população.

Impacto da sessão de hemodiálise na força de preensão manual

Resumo Introdução: A força de preensão manual (FPM) é um método simples, confiável e com bom valor preditivo para avaliar a função muscular de pacientes submetidos à hemodiálise (HD). Porém, ainda não existe um consenso a respeito do momento mais apropriado para a aferição da medida, já que o desempenho da FPM pode ser influenciado pelas flutuações hidroeletrolíticas e de pressão arterial que acometem esses pacientes. Objetivo: Investigar o impacto da sessão de diálise sobre a FPM em pacientes submetidos à HD. Métodos: Trata-se de um estudo transversal com 156 pacientes [57,7% homens, idade mediana de 56,5 (42-67) anos, 28,8% diabéticos, IMC médio de 24,75 ± 4,5 kg/m2 e tempo em HD de 38 (19,25-72,75) meses]. Foram realizadas aferições da FPM com um dinamômetro nos minutos iniciais e ao término da sessão de HD. Os valores obtidos foram comparados com um padrão de referência nacional. Dados clínicos, demográficos e laboratoriais foram coletados do prontuário médico. Resultados: Foi observada uma redução significante da FPM após a sessão de HD (28,6 ± 11,4 kg para 27,7 ± 11,7 kg; p < 0,01). A prevalência de pacientes com FPM abaixo do percentil 30 aumentou de 44,9% para 55,1% (p < 0,01). A redução da pressão arterial durante a diálise foi o único fator que se associou com a redução da FPM. Conclusão: Os achados mostram que o processo de HD influencia negativamente a FPM.

Manual de preguntas frecuentes ¿Como conseguir y mantener a tus clientes?

Bogotá Emprende

Manual sobre el comercio de servicios en los acuerdos de libre comercio negociados por Colombia

Gestión del conocimiento

Documento Matriz. Descubre tu perfil emprendedor. Manual competencias emprendedoras

Desarrollo empresarial y creación de empresa