Papel da proteína de reparo XPC na regulação das proteínas de reparo APE1, OGG1 e PARP-1 em células humanas e de camundongos

studies using UV as a source of DNA damage. However, even though unrepaired UV-induced DNA damages are related to mutagenesis, cell death and tumorigenesis, they do not explain phenotypes such as neurodegeneration and internal tumors observed in patients with syndromes like Xeroderma Pigmentosum (XP) and Cockayne Syndrome (CS) that are associated with NER deficiency. Recent evidences point to a role of NER in the repair of 8-oxodG, a typical substrate of Base Excision Repair (BER). Since deficiencies in BER result in genomic instability, neurodegenerative diseases and cancer, it was investigated in this research the impact of XPC deficiency on BER functions in human cells. It was analyzed both the expression and the cellular localization of APE1, OGG1 e PARP-1, the mainly BER enzymes, in different NER-deficient human fibroblasts. The endogenous levels of these enzymes are reduced in XPC deficient cells. Surprisingly, XP-C fibroblasts were more resistant to oxidative agents than the other NER deficient fibroblasts, despite presenting the highest of 8-oxodG. Furthermore, subtle changes in the nuclear and mitochondrial localization of APE1 were detected in XP-C fibroblasts. To confirm the impact of XPC deficiency in the regulation of APE1 and OGG1 expression and activity, we constructed a XPC-complemented cell line. Although the XPC complementation was only partial, we found that XPC-complemented cells presented increased levels of OGG1 than XPC-deficient cells. The extracts from XPC-complemented cells also presented an elevated OGG1 enzimatic activity. However, it was not observed changes in APE1 expression and activity in the XPCcomplemented cells. In addition, we found that full-length APE1 (37 kDa) and OGG1- α are in the mitochondria of XPC-deficient fibroblasts and XPC-complemented fibroblasts before and after induction of oxidative stress. On the other hand, the expression of APE1 and PARP-1 are not altered in brain and liver of XPC knockout mice. However, XPC deficiency changed the APE1 localization in hypoccampus and hypothalamus. We also observed a physical interaction between XPC and APE1 proteins in human cells. In conclusion, the data suggest that XPC protein has a role in the regulation of OGG1 expression and activity in human cells and is involved mainly in the regulation of APE1 localization in mice. Aditionally, the response of NER deficient cells under oxidative stress may not be only associated to the NER deficiency per se, but it may include the new functions of NER enzymes in regulation of expression and cell localization of BER proteins

Efeito do Fucoidam de Fucus vesiculosus em um modelo experimental de artrite reumatóide

Rheumatoid arthritis (RA) is systemic auto imune disorder. It is caracterized by chronic inflammation of joints leading to progressive erosion of cartilage and bone. We investigated the effect of the administration of fucoidan, sulfated polysaccharides, from algae Fucus vesiculosus in the acute (6h) in zymosan-induced arthritis (AZy). Wistar rats (180-230 g) were used for all groups experimental. Non-treated animals received just intraarticular injection of 1 mg the zymosan, control group received intraarticular injection of 50 µL the saline, groups received either fucoidan of Fucus vesiculosus (15, 30, 50 or 70 mg/Kg) or parecoxib (1 mg/Kg) 1 hour after injection of zymosan. After 6 h, the articular exudates were collected for evaluation of the cell influx and nitrite (Griess reaction) release. The sinovial membranes and articular cartilages were excised for histopathological analysis and by determination of the glycosaminoglycan (GAG), respectively. ZyA led to increased NO and cell influx into the joints. Therapeutic administration of the fucoidan or parecoxib did significantly inhibited the cell influx and the synovitis, as compared to non-treated rats (p<0,05), though being able to reduced NO release. Representative agarose gel electrophoresis of the GAGs, the content of condroitin-sulphate was observed during the process. These findings suggest that the fucoidan from Fucus vesiculosus has potential anti-inflammatory activity

Efeito de preparações de produtos naturais (cravo da índia e tomate) e de um produto sintético na marcação de constituintes sangüineos com tecnécio-99m e na morfologia de hemácias isoladas de sangue de ratos wistar

There are strong interests in the evaluation of the biological effects of natural and synthetic products. Blood constituents labeled with technetium-99m (99mTc) are used in nuclear medicine. The aim of this work was to study the effects of Clove (Caryophyllus aromaticas L.) and OZE (preparation used in the Health Sciences) on the labeling blood constituents with 99mTc and on the morphologic red blood cells (RBC) and the action of an extract of tomato (TO) on the labeling of blood constituents Blood samples were incubated with clove or OZE or TO, stannous chloride and 99mTc. Plasma (P), blood cells (BC), insoluble fractions (IF) of plasma and blood cells were separated. The radioactivity was counted and percentage of radioactivity (%ATI) to each blood fraction was calculated. The shape and morphometric parameter (perimeter/area ratio) were evaluated in the studies with clove and OZE. Clove extract and OZE altered significantly (p<0.05) the %ATI of blood constituents and the shape of red blood cells. However, clove extract not altered the red blood cells perimeter/area ratio. The tomato extract used at the highest concentrations reduced significantly (p<0.05) the %ATI in IF-P, although this extract did not modify the radiolabeling on BC, neither the radioactivity fixation on IFBC. The results indicate that these chemical compounds would have oxidative/chelating actions

Hiperidrose compensatória após simpatectomia toracoscópica: características, incidência e influência na satisfação do paciente

Hyperhidrosis is an idiopathic condition characterized by excessive sweating. Symptoms generally begin in childhood or early adolescence, and rarely improve with age. The excessive localized sweating generally occurs either spontaneously, or in association with stressful or emotionally charged situations. This prospective study aimed to investigate predictive factors for compensatory hyperhidrosis after thoracoscopic sympathicotomy. From 2000 to 2002, 80 patients (53 female and 27 male) underwent hyperhidrosis surgery. The patients, ranging from 12 to 56 years old, were studied and followed-up for 42.51 ±5.98 months. A satisfaction grading using a visual analogue scale -VAS (0 = not at all satisfied, and 10 = fully satisfied) was used. The surgical procedure was performed bilaterally on the second ganglion (T2) for facial hyperhidrosis, on the third and fourth ganglia (T3 and T4) for axillary hyperhidrosis, and on the third ganglion (T3) for palmar hyperhidrosis. The results showed that, 68 patients (85%) presented with compensatory sweating (CS), which was classified as mild (33.8%), moderate (33.8%) and severe (32.4%). Considering the final surgical results, 70 patients (87.5%) were satisfied with the outcome of the operation, while 10 patients (12.5%) were dissatisfied. Degrees of satisfaction varied according to sex, age, BMI and extent of denervation. Moreover, the compensatory hyperhidrosis was more severe in abdomen and back than in legs. In conclusion, although CS is a frequent adverse effect of sympathicotomy, the degree of patient satisfaction was high. Some factors were related to the occurrence and severity of CS and the most adequate patients to be submitted to this operation are young adult women whose BMI is less than 24.9

Evaluation of the Genotoxicity of Chitosan Nanoparticles for Use in Food Packaging Films

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

Effect of intermittent feeding on growth in early juveniles of the crayfish Cherax quadricarinatus

Three experiments were conducted with juveniles of the crayfish Cherax quadricarinatus to investigate the effect of intermittent feeding regimes on growth and the ability to tolerate the shortage of food. In experiment 1, stage III juveniles were assigned to one of seven intermittent feeding groups (from FS1: 1 day fed/1 day non-fed to FS7: 7 days fed/7 days non-fed) and two control groups, continuously fed (CF) and continuously starved (CS) animals; this experiment comprised a short-term intermittent feeding period until the first molt, followed by a continuous feeding period. In the experiment 2, stage III juveniles were assigned to one of three intermittent feeding groups (FS2 to FS4) and one control group (CF); it consisted of a prolonged intermittent feeding period, until the end of the experiment In the experiment 3, stage VI and VII juveniles were assigned to one of three intermittent feeding groups (FS2 to FS4) and one control (CF); it also consisted of a prolonged intermittent feeding period. The red claw crayfish juveniles were able to tolerate periods of intermittent feeding and underwent compensatory growth after continuous feed was re-established. The ability of crayfish to tolerate intermittent feeding was influenced by developmental stage and duration of the intermittent feeding period. Stage III juveniles survived, but decreased growth, when subjected to prolonged intermittent feeding. However, they showed full compensatory growth when the intermittent feeding period was short and followed by continuous feeding. on the other hand, stage VI-VII tolerated 60 days of prolonged intermittent feeding without any change in growth and survival. The hepatosomatic index (based on wet weight) values of the treatments and the control were similar, suggesting that intermittent feeding may not be considered a nutritional stress condition. The relative pleon weight (based on wet weight) values of the treatments and control were similar suggesting low use of nutrients from the muscle to increase the chance for survival. The juveniles of C quadricarinatus can tolerate relatively long periods of low food availability and this is an important adaptation for their survival in changing/unpredictable environments and an attribute favorable for the production of the species. (C) 2011 Elsevier B.V. All rights reserved.

Efeitos de um extrato aquoso de porangaba (Cordia salicifolia) na marcação de constituintes sanguíneos com tecnécio-99m e na morfologia de hemácias

Effects of a Cordia salicifolia (porangaba) extract on the labeling of blood cells (BCs) with technetium-99m ((99m)Tc) and on the morphology of red BCs were evaluated. Labeling of cellular and molecular structures with (99m)Tc depends on a reducing agent. Some physical characteristics, as visible absorbance spectrum, electric conductivity, and refractive index of this porangaba extract, were also determined. Blood samples from Wistar rats were incubated with porangaba extract or with 0.9% NaCl (control). Labeling of blood constituents with (99m)Tc was performed. Plasma (P) and BCs, both soluble (SF-P and SF-BC) and insoluble (IF-P and IF-BC) fractions, were separated. The radioactivity in each fraction was counted, and the percentage of radioactivity incorporated (%ATI) was calculated. Blood smears were prepared, fixed, and stained, and the morphology of the red BCs was evaluated. Data showed an absorbance peak at 480 nm and electric conductibility and refractive index concentration-dependent. Porangaba extract decreased significantly (P < .05) the BC, IF-P, and IF-BC %ATI, and no modifications were verified on the shape of red BCs. Analysis of the results reveals that some physical parameters could be useful to aid in characterizing the extract studied. Moreover, it is possible that chemical compounds of this extract could have chelating/redox actions or be capable of binding to plasma and/or cellular proteins

Efeito de um extrato de Ganoderma lucidum (Reishi) na marcação de constituintes sanguíneos com tecnécio-99M e na sobrevivência de Escherichia coli

Clinical evaluations have been made possible with radiobiocomplexes marked with tecnecium-99m (99mTc). Natural or synthetic drugs are able to interfere in the marking of blood structures with 99m Tc. Also, the toxicity of several natural products has been described. The aim of this study was evaluating the effect of an extract of Ganoderma lucidum (Reishi) in the marking of blood constituents with 98m Tc and in the survival of Escherichia coli. Blood samples from Wistar rats were treated with reishi extract. Radiomarking procedure was performed. Samples of plasma (P), blood cells (CS), and insoluble (FI) and soluble (FS) fractions of P and CS were separated and the radioactivity was counted to determine radioactivity percentages (%ATI). Escherichia coli AB1157 cultures were treated with stannous chloride in the presence and absence of the reishi extract. Blood samples and bacterial cultures treated with NaCl 0.9% were used as controls. Data indicated that the reishi extract has significantly altered (p<0,05) the %ATI of P, CS, FI-P, FS-P, FI-CS e FS-CS, as well as it has increased survival of bacterial cultures treated with stannous chloride. Our results suggest that the Reishi extract would be able to present a redox/ chelant action by altering blood constituent marking with 99mTc and by protecting bacterial cultures against stannous chloride-induced oxydating lesions. The study had a multidisciplinary character, with the participation of the following areas of knowledge: Biophysics, Radiobiology, Botanics, Phytotherapy, and Hematology

Digestão total e parcial de alguns componentes de dietas contendo diferentes níveis de casca de soja e Fontes de nitrogênio, em bovinos

Este experimento foi realizado para avaliar o efeito de dois níveis de casca de soja (CS) e de três fontes de nitrogênio sobre a digestão total e parcial aparentes no rúmen e nos intestinos. Seis novilhos Holandês x Zebu com idade média de 12 meses e 229 kg de PV foram fistulados no rúmen e no abomaso e distribuídos em quadrado latino 6x6 e alimentados individualmente ad libitum. O consumo e as digestibilidades totais e parciais dos nutrientes foram avaliados. Não houve efeito da fonte de proteína sobre o consumo, quantidades de matéria seca (MS), matéria orgânica (MO), proteína bruta (PB), fibra em detergente neutro (FDN) e da fibra em detergente ácido (FDA) no rúmen, no abomaso e nas fezes e sobre os coeficientes de digestibilidades destes componentes. O aumento, porém, do nível de casca de soja elevou os coeficientes de digestibilidade de todos os componentes estudados nos locais do trato digestivo, com exceção nos intestinos, onde o nível de casca de soja não afetou a digestão da PB, FDN e da FDA. Os resultados indicam que a casca de soja pode substituir tanto o milho como, parcialmente, a fração volumosa nas rações de ruminantes. A farinha de subproduto avícola e a amiréia podem substituir o farelo de soja nas rações de bovinos sem reflexos negativos sobre a digestão de seus componentes nutritivos.

Eficácia do herbicida glifosato-potássico submetido à chuva simulada após a aplicação

Apesar de possuir boa ação herbicida, alguns fatores afetam a eficácia do glifosato e, nesse contexto, realizaram-se estudos buscando formulações mais eficazes. O experimento foi instalado em área do Campus da FCAV-UNESP, Jaboticabal (SP). Testou-se a formulação potássica de glifosato comparada com solução aquosa e grânulos dispersíveis em água à mesma dosagem (0,36 kg.ha-1 e.a) em cinco intervalos (1, 2, 4, 6 e > 48 horas) sem chuva após a aplicação, comparados com uma testemunha sem uso de herbicidas. Foram aplicados em pós-emergência sobre Brachiaria plantaginea, B. decumbens e Digitaria horizontalis. A aplicação foi feita com pulverizador costal a CO2, à pressão de 2,5 kg.cm-2, munido de barra com seis bicos DG 110.02 e consumo de calda equivalente a 200 L.ha-1. Realizaram-se cinco avaliações de porcentagem de controle em relação à testemunha infestada, atribuídas visualmente, até 42 dias após aplicação. Notou-se que o controle da formulação potássica sobre as espécies avaliadas foi mais precoce, necessitando apenas quatro horas após sua aplicação para se igualar ao intervalo superior a 48 horas sem chuva. As demais necessitaram de, no mínimo, seis horas de intervalo.

Eficácia de uma nova formulação de glifosato para o controle de grama-seda (Cynodon dactylon), em pomar de citros.

Avaliou-se a eficácia do herbicida glifosato potássico, comparado ao sulfosato e ao glifosato amínico, utilizados como padrões comerciais. Os herbicidas foram aplicados em um pomar de laranja da variedade Pêra-rio, infestado por grama-seda (Cynodon dactylon) com aproximadamente 25 cm de altura, em 70% da área. O experimento foi instalado no município de Taiúva-SP, em solo Podzólico Vermelho-Amarelo, distrófico, de textura arenosa, com relevo suave ondulado. Os tratamentos testados foram: glifosato potássico (ZAPP Qi) a 1,05; 2,10 e 2,50 kg e.a./ha, sulfosato (ZAPP) e glifosato amínico (ROUNDUP CS) a 1,98 e 2,16 kg e.a./ha, respectivamente, além da testemunha sem capina. Os herbicidas foram aplicados em pós-emergência da planta daninha, com pulverizador costal, à pressão constante (mantida por CO2 comprimido) de 2,5 kg/cm², munido de barra com seis bicos de jato plano (leque) DG 11002, com um consumo de calda equivalente a 200 L/ha. Os melhores resultados de controle foram obtidos com 2,5 kg e.a./ha do glifosato potássico, que também garantiu menor brotação da grama-seda. No entanto, a dosagem de 2,10 kg e.a./ha deste mesmo herbicida não foi menos eficaz que o sulfosato e o glifosato, com 1,98 e 2,16 kg e.a./ha, respectivamente.

Efeito da adição de óleos ao acaricida cyhexatin sobre o ácaro Brevipalpus phoenicis e na retenção de calda por folhas de citros

A adição de óleos à calda de pulverização, muitas vezes, é utilizada a campo sem o adequado conhecimento sobre a absorção do produto fitossanitário pelo alvo, retenção de calda e até mesmo sobre a praga e a cultura. O objetivo do trabalho foi avaliar o efeito da adição de óleos ao acaricida cyhexatin sobre o ácaro Brevipalpus phoenicis e na retenção de calda por folhas de citros. Avaliou-se a mortalidade de ácaros, utilizando-se de frutos de laranja com uma arena circundada com cola entomológica para confinar os ácaros. Adotou-se o delineamento inteiramente casualizado, em esquema fatorial, constituído pelos fatores: duas formulações de cyhexatin (WG e SC), dois tipos de óleo (mineral e vegetal) e duas concentrações dos óleos (0,5 e 1,0%), e mais dois tratamentos adicionais (acaricidas não adicionados de óleo) e uma testemunha sem aplicação. A aplicação dos produtos foi realizada sobre frutos de laranja até além do ponto de escorrimento. Logo após a aplicação, transferiram-se 10 ácaros B. phoenicis para cada fruto.A contagem dos ácaros vivos, mortos e retidos na barreira adesiva foi realizada um dia após a aplicação. Para a determinação da quantidade de calda retida, utilizaram-se folhas de laranjeira, que foram pulverizadas até além do ponto de escorrimento, adotando-se os mesmos tratamentos e o delineamento estatístico mencionados para a avaliação da mortalidade de ácaros, com exceção da testemunha sem aplicação. Determinou-se a massa de líquido retido após a aplicação dos produtos por folha, com auxílio de balança de precisão. Verificou-se que um dia após a aplicação dos produtos, todos os tratamentos apresentaram mortalidade de B. phoenicis acima de 99%. Dessa forma, a adição de óleo, seja mineral, seja vegetal, ao acaricida cyhexatin não afetou a eficácia biológica deste acaricida nas formulações SC e WG. A maior fuga de B. phoenicis para a barreira de cola foi verificada nos tratamentos com adição de óleos, em comparação ao cyhexatin aplicado isoladamente. A adição de óleo, seja mineral, seja vegetal, ao cyhexatin na calda de pulverização reduziu a quantidade máxima de líquido retido pelas folhas de citros, podendo contribuir para a redução da quantidade de calda necessária para uma boa cobertura da planta.

Gonadal steroids levels and vitellogenesis in the formation of oocytes in Prochilodus lineatus (Valenciennes) (Teleostei: Characiformes)

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

Comparison of microbial numbers in soils by using various culture media and temperatures

The influence of different media and incubation temperatures on the quantification of microbial populations in sorghum, eucalyptus and forest soils was evaluated. Microbial growth was compared by using complex (tryptone soybean agar, TSA, casein-starch, CS, and Martin) and saline (Thorton, M3, Czapeck) media and incubation temperatures of 25 and 30&DEG; C. Higher numbers of total bacterial. and fungal colony-forming units (CFU) were observed in sorghum soils, and of spore-forming and Gram-negative bacteria in forest soils than other soils. Actinomycetes counts were highest in forest soil when using CS medium at 30&DEG; C and in sorghum soil at 25&DEG; C in M3 medium. Microorganism counts were dependent on the media and incubation temperatures. The counts at temperatures of 30&DEG; C were significantly higher than at 25&DEG; C. Microbial quantification was best when using TSA medium for total. and spore-forming bacteria, Thorton for Gram-negative bacteria, M3 for actinomycetes, and Martin for fungi. © 2005 Elsevier GmbH. All rights reserved.

Modelos para estimativa do subfator cobertura-manejo (CiII) relativo à erosão entressulcos

A análise de um modelo de erosão é um crítico passo no desenvolvimento de uma ferramenta de predição da erosão aplicável e válida; isso é crucial para avaliar o desempenho dos modelos existentes para assegurar que as estimativas de um modelo condizem com a realidade. O objetivo do presente trabalho foi avaliar modelos para a predição do subfator cobertura e manejo (CiII) relativo à erosão entressulcos. Um experimento fatorial completo foi conduzido com cinco doses de resíduo de milho (0; 0,05; 0,15; 0,40 e 0,80 kg m-2), quatro declives e duas repetições, sob condições de prévio umedecimento para determinar as taxas de erosão entressulcos (Di) e enxurrada (R). Num primeiro experimento, foi avaliada a erodibilidade entressulcos (Ki) e o subfator cobertura e manejo (CiII), em parcelas experimentais de 0,5 x 0,75 m, em solo recentemente preparado. Num segundo experimento, foram avaliados Di, R, Ki e CiII, também em parcelas de 0,5 x 0,75 m, em solo recém- preparado. Os valores de Di, R, Ki e CiII, obtidos no segundo experimento, foram utilizados na avaliação dos modelos testados. Os modelos CiII = e-2,50 CS/100 e CiII = e -2,238 CS/100 apresentaram boas estimativas para o subfator CiII.