Cloning, expression, molecular modelling and docking analysis of glutathione transferase from Saccharum officinarum

Sugarcane yield and quality are affected by a number of biotic and abiotic stresses. In response to such stresses, plants may increase the activities of some enzymes such as glutathione transferase (GST), which are involved in the detoxification of xenobiotics. Thus, a sugarcane GST was modelled and molecular docked using the program LIGIN to investigate the contributions of the active site residues towards the binding of reduced glutathione (GSH) and 1-chloro-2,4-dinitrobenzene (CDNB). As a result, W13 and I119 were identified as key residues for the specificity of sugarcane GSTF1 (SoGSTF1) towards CDNB. To obtain a better understanding of the catalytic specificity of sugarcane GST (SoGSTF1), two mutants were designed, W13L and I119F. Tertiary structure models and the same docking procedure were performed to explain the interactions between sugarcane GSTs with GSH and CDNB. An electron-sharing network for GSH interaction was also proposed. The SoGSTF1 and the mutated gene constructions were cloned and expressed in Escherichia coli and the expressed protein purified. Kinetic analyses revealed different Km values not only for CDNB, but also for GSH. The Km values were 0.2, 1.3 and 0.3 mM for GSH, and 0.9, 1.2 and 0.5 mM for CDNB, for the wild type, W13L mutant and I119F mutant, respectively. The V(max) values were 297.6, 224.5 and 171.8 mu mol min(-1) mg(-1) protein for GSH, and 372.3, 170.6 and 160.4 mu mol min(-1) mg(-1) protein for CDNB.

Deficiências de kacronutrientes e de boro em seringueira (Hevea brasiliensis L.)

Plantas de seringueira (Hevea brasiliensis L.) foram cultivadas em casa de vegetação, em quartzo moído, irrigado com soluções nutritivas, e submetidas aos seguintes tratamentos: completo, omissão de N, omissão de P, omissão de Ca, omissão de Mg, omissão de S e omissão de B, com o objetivo de: (a) obter sintomas de deficiências de macronutrientes e de boro; (b) analisar o crescimento das plantas através da produção de matéria seca; (c) determinar a concentração de macro e micronutrientes nas folhas, caule e raízes das plantas cultivadas nos diversos tratamentos. Os sintomas visuais de deficiência foram identificados e descritos. As plantas foram coletadas e separadas em raiz, caule e folhas, e determinaram-se os teores de macro e micronutrientes . Os resultados mostraram: - foram identificados sintomas de deficiências para todos os tratamentos com omissão de nutrientes (N, P, K, Ca, Mg, S e B); - a omissão de N, K, Mg ou B da solução nutritiva diminuiu o crescimento das plantas; - as concentrações dos elementos nas folhas de plantas com sintomas e sem sintomas de deficiência foram, respectivamente: N% = 1,94 e 3,40; P% = 0,14 e 0,25; K% = 0,79 e 2,22; Ca% = 0,59e 1,28; Mg% = 0,26 e 0,50; S% = 0,10 e 0,10; Bppm = 31 ,3 e 171,8.

On blame-freeness and reciprocity: an experimental study

The theory of reciprocity is predicated on the assumption that people are willing to reward nice or kind acts and to punish unkind ones. This assumption raises the question as to how to define kindness. In this paper we offer a new definition of kindness that we call “blame-freeness.” Put most simply, blame-freeness states that in judging whether player i has been kind or unkind to player j in a social situation, player j would have to put himself in the strategic position of player i, while retaining his preferences, and ask if he would have acted in a manner that was worse than i did under identical circumstances. If j would have acted in a more unkind manner than i acted, then we say that j does not blame i for his behavior. If, however, j would have been nicer than i was, then we say that “j blames i” for his actions (i’s actions were blameworthy). We consider this notion a natural, intuitive and empirically relevant way to explain the motives of people engaged in reciprocal behavior. After developing the conceptual framework, we then test this concept in a laboratory experiment involving tournaments and find significant support for the theory.

Hume : tratado sobre la naturaleza humana : preguntas y respuestas al texto.

Se presenta un instrumento pedagógico para que el alumnado, de COU y Selectividad entienda la filosofía a través de la obra de sus autores (en este caso se estudia a Hume). El esquema seguido es el siguiente: introducción que sitúa el texto dentro de la obra del autor; vocabulario de conceptos clave para la interpretación del texto; división y clasificación de las partes del texto, para su mayor clarificación, y plateamiento de preguntas y propuestas de respuestas.

Cinética do transporte jejunal de glicose em ratos com deficiência de niacina.

The kinetic of jejunal glucose transport was studied using perfused rat jejunum in vivo. Ninety rats were fed a diet deficient in niacin and 90 a control diet. The jejunal loops of 7 groups of animal were infused each group with one of following solutions of glucose: 5, 10, 20, 40, 80, 160 and 300 mM/l. The Vmax and Km values were determined. The results showed that the vitamin-deficient rats absorbed less glucose independently of the amount infused and these animals had lower Vmax (133.7 microM/15 min/15 cm) and Km (192.1 mM/l) than control groups (294.1 microM/15 min/15 cm and 171.8 mM/l, respectively). In conclusion one can assume that niacin deficiency leads to a decreased glucose absorption in the jejunal loops, when tested as in our experimental model.

A comunidade perifítica e suas relações com a qualidade da água no reservatório de Itupararanga (SP, Brasil)

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Brain activation during craving for alcohol measured by positron emission tomography

OBJECTIVE: Craving for alcohol is probably involved in acquisition and maintenance of alcohol dependence to a substantial degree. However, the brain substrates and mechanisms that underlie alcohol craving await more detailed elucidation. METHOD: Positron emission tomography was used to map regional cerebral blood flow (CBF) in 21 detoxified patients with alcohol dependence during exposure to alcoholic and non-alcoholic beverages. RESULTS: During the alcohol condition compared with the control condition, significantly increased CBF was found in the ventral putamen. Additionally, activated areas included insula, dorsolateral prefrontal cortex and cerebellum. Cerebral blood flow increase in these regions was related to self-reports of craving assessed in the alcoholic patients. CONCLUSIONS: In this investigation, cue-induced alcohol craving was associated with activation of brain regions particularly involved in brain reward mechanisms, memory and attentional processes. These results are consistent with studies on craving for other addictive substances and may offer strategies for more elaborate studies on the neurobiology of addiction.

Performance characteristics of automotive engines in the United States. Second series--report no. 7. 1977 Ford 171 CID (2.8 liters), 2V. Interim report.

Transportation Systems Center, Cambridge, Mass.

Fragm. lat. III 8 - Digestum novum (De regulis iuris D.50.17.65 - 117; 158 - 169; 171 - 192)

Trägerbände: Ms. Barth. 52; Ms. Barth. 66; Vorbesitzer: St. Peter Urach; Bartholomaeusstift Frankfurt am Main

The Monocarboxylate Transporter 8 and L-Type Amino Acid Transporters 1 and 2 Are Expressed in Mouse Skeletons and in Osteoblastic MC3T3-E1 Cells

Background: Several plasma membrane transporters have been shown to mediate the cellular influx and/or efflux of iodothyronines, including the sodium-independent organic anion co-transporting polypeptide 1 (OATP1), the sodium taurocholate co-transporting polypeptide (NTCP), the L-type amino acid transporter 1 (LAT1) and 2 (LAT2), and the monocarboxylate transporter 8 (MCT8). The aim of this study was to investigate if the mRNAs of these transporters were expressed and regulated by thyroid hormone (TH) in mouse calvaria-derived osteoblastic MC3T3-E1 cells and in the fetal and postnatal bones of mice. Methods: The mRNA expression of the iodothyronine transporters was investigated with real-time polymerase chain reaction analysis in euthyroid and hypothyroid fetuses and litters of mice and in MC3T3-E1 cells treated with increasing doses of triiodothyronine (T(3); 10(-10) to 10(-6) M) or with 10(-8) M T(3) for 1-9 days. Results: MCT8, LAT1, and LAT2 mRNAs were detected in fetal and postnatal femurs and in MC3T3-E1 cells, while OATP1 and NTCP mRNAs were not. LAT1 and LAT2 mRNAs were not affected by TH status in vivo or in vitro or by the stage of bone development or osteoblast maturation (analyzed by the expression of osteocalcin and alkaline phosphatase, which are key markers of osteoblastic differentiation). In contrast, the femoral mRNA expression of MCT8 decreased significantly during post-natal development, whereas MCT8 mRNA expression increased as MC3T3-E1 cells differentiated. We also showed that MCT8 mRNA was up-regulated in the femur of hypothyroid animals, and that it was down-regulated by treatment with T(3) in MC3T3-E1 cells. Conclusions: This is the first study to demonstrate the mRNA expression of LAT1, LAT2, and MCT8 in the bone tissue of mice and in osteoblast-like cells. In addition, the pattern of MCT8 expression observed in vivo and in vitro suggests that MCT8 may be important to modulate TH effects on osteoblast differentiation and on bone development and metabolism.