Sistema antioxidante envolvendo o ciclo metabólico da glutationa associado a métodos eletroanalíticos na avaliação do estresse oxidativo

The most relevant advances on the analytical applications of glutathione determination based on glutathione redox cycle and the antioxidant system are given. The main enzymes that participate of the glutathione metabolism are the glutathione peroxidase and glutathione reductase. The glutathione peroxidase has a major role in the removal of hydrogen peroxide and lipid peroxides from the cells. These enzymes, operating in tandem with catalase and superoxide dismutase promote a scavenging of oxyradical products in tissues minimizing damages caused by these species. Reduced glutathione is the major intracellular thiol found in mammals and changes in the glutathione concentration in biological fluids or tissues may provide a useful marker in certain disorders like hemolytic anemia, myocardial oxidative stress and in the investigation of some kinds of cancers. Particular attention is devoted to the main advantages supplied by biosensors in which there is an incorporation of bioactive materials for the glutathione determination. The correlation between stability and sensitivity of some reduced glutathione electrochemical sensors is discussed.

A Gendered Voice in Translation: Translating Like a Feminist

This paper addresses some of the challenges inherent in finding and showing a gendered voice in translation. The starting point is my own experience as a feminist translator of both feminist and non-feminist texts. Textual practices like translating necessarily interact with current theoretical debates. In turn, theoretical writing on feminism enriches and informs one’s translating activity. This interplay between theoretical models and textual practices was particularly made evident to me as I rendered Essentially speaking, by Diana Fuss, into Catalan. In this article I intend to transcend anecdotes of translating individual texts and consider how translating equals rewriting oneself; it involves rethinking writing practices. I will specifically address the rethinking of (1) one’s identity when translating ‘like’ a feminist, (2) performativity in gender and in translation, and (3) agency and (In)visibility.

Catalase in fluvial biofilms: a comparison between different extraction methods and example of application in a metal-polluted river

Antioxidant enzymes are involved in important processes of cell detoxification during oxidative stress and have, therefore, been used as biomarkers in algae. Nevertheless, their limited use in fluvial biofilms may be due to the complexity of such communities. Here, a comparison between different extraction methods was performed to obtain a reliable method for catalase extraction from fluvial biofilms. Homogenization followed by glass bead disruption appeared to be the best compromise for catalase extraction. This method was then applied to a field study in a metal-polluted stream (Riou Mort, France). The most polluted sites were characterized by a catalase activity 4–6 times lower than in the low-polluted site. Results of the comparison process and its application are promising for the use of catalase activity as an early warning biomarker of toxicity using biofilms in the laboratory and in the field

Estudo de liberação e permeação in vitro do diclofenaco de dietilamônio em microemulsão gel-like

The goal of this study was to produce and characterize a new microemulsion gel-like carrier system (MEG) by using the pseudo-ternary phase-diagram concept. The diclofenac diethylamine (DDA) was incorporated in the MEG and its in vitro release and permeation profiles were performed using Franz-type diffusion cells. The results revealed that the commercial DDA emulgel provided significantly higher Kp of DDA (2.2-fold) as compared to the MEG. Similar data were obtained in the permeation studies in which DDA Kp 4.7-fold higher. Therefore, MEG presents higher potential as a topical delivery system for DDA when compared to the commercial DDA emulgel.

Anxiolytic-like effects of erythrinian alkaloids from erythrina suberosa

Two alkaloids, erysodine (1) and erysothrine (2) were isolated from the flowers of a Pakistani medicinal plant, Erythrina suberosa. These compounds were investigated for anxiolytic properties, and the results showed significant effect, in an acute oral treatment with 1-2, which were suspended in saline (NaCl 0.9%) plus DMSO 1%, and evaluated in 122 Swiss male mice exposed to two tests of anxiety - the elevated plus-maze (EPM) and the light/dark transition model (LDTM).

Atividade eletrocatalítica de sistemas biomiméticos da enzima catalase

In this work, we describe the immobilization of the dinuclear compound [Cu2(apyhist)2Cl2](ClO4)2 (1) and its derived cations complexes, obtained in water solution or by deprotonation of the imidazolate moiety in the ligand leading to a cyclic tetranuclear species, in the Nafion® membrane on glass carbon electrode surface. After that, we studied the influence of the equilibrium in the electrocatalytic activity towards the reduction of H2O2 in the development of an amperometric sensor for the analytical determination of hydrogen peroxide. This strategy proved successful, and the electrochemical behaviour of the all complexes formed within the Nafion® coatings was characterized. We also provide evidence that its related cyclic tetranuclear imidazolate-bridged complex acts as a catalysts for the intramolecular, two-electron reduction of H2O2.

PCR amplification and sequence analyses of reverse transcriptase-like genes in Crinipellis perniciosa isolates

Reverse transcriptase (RT) sequence analysis is an important technique used to detect the presence of transposable elements in a genome. Putative RT sequences were analyzed in the genome of the pathogenic fungus C. perniciosa, the causal agent of witches' broom disease of cocoa. A 394 bp fragment was amplified from genomic DNA of different isolates of C. perniciosa belonging to C-, L-, and S-biotypes and collected from various geographical areas. The cleavage of PCR products with restriction enzymes and the sequencing of various RT fragments indicated the presence of several sequences showing transition events (G:C to A:T). Southern blot analysis revealed high copy numbers of RT signals, forming different patterns among C-, S-, and L-biotype isolates. Sequence comparisons of the predicted RT peptide indicate a close relationship with the RT protein from thegypsy family of LTR-retrotransposons. The possible role of these retrotransposons in generating genetic variability in the homothallic C. perniciosa is discussed.

Accelerated ageing like a vigour test to diasporas of Myracrodruon urundeuva Fr. Allem

This work established the adequate temperature and the period of exposition for the accelerated ageing test with diasporas of Myracrodruon urundeuva from an area of the Cerrado in the state of Mato Grosso, Brazil. Before and after the ageing, at the temperatures of 40ºC, 41ºC, 42ºC and 45ºC combined with periods of 12, 18, 24, 30 and 36 hours, the water content and germination were evaluated. For each treatment, 12g of diasporas in mini chambers with 40 mL of distilled water were submitted to the accelerated ageing test. A total of 100 diasporas, divided into four replications, in plastic boxes, on two sheets of blotting paper, in germinator at 25ºC and 8 hours of photoperiod were germinated. In all treatments the content of water of the aged diasporas was superior to 20% and this value stabilized itself between 25% and 28%, from the 18 hours of exposition. Independently of the tested periods, the ageing at the temperatures of 40ºC, 41ºC and 42ºC did not affect the germination. After the ageing at 45ºC, the germination did not differ among the periods of 12, 18 and 24 hours of exposition, but in all these periods it was inferior to the control and superior to the periods of 30 and 36 hours. In these last two periods, fungi were observed. The accelerated ageing of diasporas of Myracrodruon urundeuva should be conducted at the temperature of 45ºC, during the exposition periods of 12 to 24 hours.

Atividade da catalase no pulmão, rim e intestino delgado não isquemiado de ratos após reperfusão intestinal

OBJETIVO: Avaliar a atividade catalase, após lesão por isquemia e reperfusão intestinal e estudar as alterações deste antioxidante em órgãos situados à distância do insulto inicial. MÉTODOS: Utilizaram-se 18 ratos do tipo Wistar, aleatoriamente distribuídos em três grupos. 1-Controle, 2-Simulação e 3-Isquemia/Reperfusão. Neste último, realizou-se isquemia no íleo, por 60 minutos, seguida de reperfusão por 30 minutos. No grupo 2 efetuou-se apenas uma laparotomia. Foram retirados, de todos os animais, segmentos do intestino com e sem reperfusão, além do pulmão e rim direitos para exame com microscopia óptica. A atividade da catalase foi aferida em espectrofotômetro ajustado para 240 nm. Utilizaram-se os testes estatísticos Mann e Whitney e Kruskal Wallis. RESULTADOS: Observou-se aumento significante (p < 0.05), da atividade da catalase nas porções do intestino isquemiado e não isquemiado, além do pulmão. Houve redução da atividade enzimática no rim. No grupo com reperfusão observaram-se alteração nas vilosidades, infiltrado inflamatório em todas as vísceras, além de áreas de atelectasia pulmonar. CONCLUSÃO: O estresse oxidativo intestinal, em ratos, causa alterações bioquímicas à distância com mobilização dos mecanismos de defesa antioxidante pulmonar, em segmento intestinal não isquemiado e no rim, com esgotamento precoce das reservas deste último, no entanto, sem lesão celular relevante, destas vísceras.

Perfil de susceptibilidade a antimicrobianos em amostras de cocos Gram-positivos, catalase negativos, isoladas de mastite subclínica bubalina

Estudou-se o perfil de susceptibilidade a antimicrobianos em cocos Gram-positivos catalase negativos (21 amostras de Lactococcus garvieae e 6 de Enterococcus gallinarum), isoladas do leite de fêmeas com mastite subclínica e pertencentes a uma população composta por seis rebanhos bubalinos localizados no Estado do Rio de Janeiro. O teste utilizado foi o da difusão de discos em agar Müller Hinton, segundo recomendações do National Committee for Clinical Laboratory Standards - NCCLS, tendo sido testados discos com ampicilina (10mg), cefalotina (30mg), cefotaxima (30mg), cefoxitina (30mg), cloranfenicol (30mg), eritromicina (15mg), gentamicina (10mg), nitrofurantoína (300mg), norfloxacina (10mg), penicilina (10 UI), tetraciclina (30mg) e vancomicina (30mg). Os resultados evidenciaram que em se tratando de Lactococcus garvieae, o antimicrobiano mais eficiente foi o nitrofurantoína com 85,71% de sensibilidade, seguido da cefotaxima (61,90%), vancomicina (52,38%), norfloxacina (47,62%) e cefalotina (47,62%). A maior resistência foi desenvolvida frente a penicilina e ampicilina, com 95,24% de resistênciapara os dois antimicrobianos testados. O perfil de susceptibilidade desenvolvido pelas amostras de Enterococcus gallinarum, mostrou baixa sensibilidade frente aos antimicrobianos testados, onde os maiores índices foram observados frente eritromicina e gentamicina, com 33,34% de sensibilidade para ambos; quanto à resistência desenvolvida, foi possível observar 100% de resistência com relação a vancomicina e tetraciclina, seguindo-se cloranfenicol, penicilina, ampicilina, cefoxitina, cefalotina, cefotaxima, norfloxacina e nitrofurantoína, todas evidenciando uma resistência de 83,33% das amostras testadas.

Animal infections by vaccinia-like viruses in the state of Rio de Janeiro: an expanding disease

In the present study we investigated the presence of infections by vaccinia-like viruses in dairy cattle from 12 counties in the state of Rio de Janeiro in the last 9 years. Clinical specimens were collected from adult animals with vesicular/pustular lesions mainly in the udder and teats, and from calves with lesions around the nose and mouth. A plaque reduction neutralization test (PRNT) was applied to search for antibodies to Orthopoxvirus; the vesicular/pustular fluids and scabs were examined by PCR, electron microscopy (EM) and by inoculation in VERO cells for virus isolation. Antibodies to Orthopoxvirus were detected in most cases. The PCR test indicated a high nucleotide homology among the isolates and the vaccinia viruses (VACV) used as controls. By EM, typical orthopoxvirus particles were observed in some specimens. The agents isolated in tissue culture were confirmed as vaccinia-like viruses by EM and PCR. The HA gene of the vaccinia-like Cantagalo/IOC virus isolated in our laboratory was sequenced and compared with other vaccinia-like isolates, showing high homology with the original Cantagalo strain, both strains isolated in 1999 from dairy cattle. Antibodies to Orthopoxvirus were detected in one wild rodent (genus Akodon sp.) collected in the northwestern region of the state, indicating the circulation of poxvirus in this area. Nonetheless, PCR applied to tissue samples collected from the wild rodents were negative. Vesicular/pustular lesions in people in close contact with animals have been also recorded. Thus, the vaccinia-like virus infections in cattle and humans in the state seem to be an expanding condition, resulting in economic losses to dairy herds and leading to transient incapacitating human disease. Therefore, a possible immunization of the dairy cattle in the state should be carefully evaluated.

Development and evaluation of a loop-mediated isothermal amplification (LAMP) assay for rapid detection of Actinobacillus pleuropneumoniae based the dsbE-like gene

This paper reports on the development and validation of a loop-mediated isothermal amplification assay (LAMP) for the rapid and specific detection of Actinobacillus pleuropneumoniae (A. pleuropneumoniae). A set of six primers were designed derived from the dsbE-like gene of A.pleuropneumoniae and validate the assay using 9 A. pleuropneumoniae reference/field strains, 132 clinical isolates and 9 other pathogens. The results indicated that positive reactions were confirmed for all A. pleuropneumoniae strains and specimens by LAMP at 63ºC for 60 min and no cross-reactivity were observed from other non-A.pleuropneumoniae including Haemophilus parasuis, Escherichia coli, Pasteurella multocida, Bordetella bronchiseptica, Streptococcus suis, Salmonella enterica, Staphylococcus, porcine reproductive and respiratory syndrome virus (PRRSV), and Pseudorabies virus. The detection limit of the conventional PCR was 10² CFU per PCR test tube, while that of the LAMP was 5 copies per tube. Therefore, the sensitivity of LAMP was higher than that of PCR. Moreover, the LAMP assay provided a rapid yet simple test of A. pleuropneumoniae suitable for laboratory diagnosis and pen-side detection due to ease of operation and the requirement of only a regular water bath or heat block for the reaction.

The influence of selected pathological states on the somatostatin-like immunoreactive (SOM-LI) endocrine cells in the mucosal layer of the porcine descending colon

This study reports on changes in the number of somatostatin-like immunoreactive (SOM-LI) endocrine cells in the porcine descending colon, caused by chemically driven inflammation, axotomy and proliferative enteropathy (PE). The distribution pattern of SOM-LI endocrine cells has been studied using the routine single-labelling immunofluorescence technique. Semi-quantitative evaluation of the number of the SOM-immunostained endocrine cells within the mucosal layer of the porcine descending colon has been based on counting of all endocrine cells immunoreactive to SOM per unit area (0,1 mm²). Under physiological conditions the number of SOM-LI endocrine cells has been shown to constitute 3,30±0,22. All applied pathological processes resulted in changes in the SOM-like immunoreactivity, which varied in particular processes studied. The number of SOM-LI endocrine cells increased to 6,28±0,31 and 4,43±0,35 during chemically driven inflammation and proliferative enteropathy, respectively, and decreased to 1,17%±0,16 after axotomy. The obtained results suggest that SOM-LI endocrine cells may participate in various pathological states within porcine descending colon and their functions probably depend on the type of pathological factor.

Porcine follicular fluid concentration of free insulin-like growth factor-I collected from different diameter ovarian follicles

The study aimed to quantify the concentrations of free IGF-I in serum and fluid of ovarian follicles in pre-pubertal gilts and describe the ovarian morphology by measuring the size of the ovaries and counting the number of surface follicles. Ovaries (n=1,000) from pre-pubertal gilts were obtained immediately after slaughter. A total of 10 samplings were performed, with ovaries obtained from 50 females for each collection. The follicles situated on the surface of each ovary were classified as small (SFs, 2 to 5mm in diameter) or large (LFs 6 to 10mm in diameter) and the follicular fluid was obtained by follicle aspiration. The collection of serum samples was performed after the gilts exsanguination using sterile tubes. From the pool of serum and follicular fluid obtained from 50 females, the concentration of free IGF-I was determined in each sample using an enzyme immunoassay kit (ELISA). The description of ovarian morphometry was performed in 100 ovaries from randomly selected gilts. The larger and smaller lengths of ovaries were measured, and the total number of SFs and LFs present on the surface of each ovary were also counted. The IGF-I concentration was greater (P<0.05) in LFs (170.92±88.29 ng/mL) compared with SFs (67.39±49.90ng/mL) and serum (73.48±34.63ng/mL). The largest and smallest length of the ovaries was 26.0±3.0 and 19.0mm ±2.0mm, respectively. The number of SFs (70.86±25.76) was greater (P<0.01) than LFs (6.54±5.26). The study concluded that LFs present greater levels of IGF-I when compared with SFs and blood, which is related to increased activity of the LFs and its differentiation to ovulation. In addition, ovaries of pre-pubertal gilts have a higher number of SFs compared to LFs. Therefore, our study demonstrated unique data regarding the physiological concentration of free IGF-I in ovarian follicles, that can be used in future research to evaluate the addition of this hormone in the in vitro production media of porcine embryos with the goal to improve the technique efficiency.