pH-Sensitive Binding of Cytochrome c to the Inner Mitochondrial Membrane. Implications for the Participation of the Protein in Cell Respiration and Apoptosis

Cytochrome c exhibits two positively charged sites: site A containing lysine residues with high pK(a) values and site L containing ionizable groups with pK(aobs),values around 7.0. This protein feature implies that cytochrome c can participate in the fusion of mitochondria and have its detachment from the inner membrane regulated by cell acidosis and alkalosis. In this study, We demonstrated that both horse and tuna cytochrome c exhibited two types of binding to inner mitochondrial membranes that contributed to respiration: a high-affinity and low-efficiency pi-I-independent binding (microscopic dissociation constant K(sapp2), similar to 10 nM) and a low-affinity and high-efficiency pH-dependent binding that for horse cytochrome c had a pK(a) of similar to 6.7. For tuna cytochrome c (Lys22 and His33 replaced with Asn and Trp, respectively), the effect of pH on K(sapp1), was less striking than for the horse heme protein, and both tuna and horse cytochrome c had closed K(sapp1) values at pH 7.2 and 6.2, respectively. Recombinant mutated cytochrome c H26N and H33N also restored the respiration of the cytochrome c-depleted mitoplast in a pH-dependent manner. Consistently, the detachment of cytochrome c from nondepleted mitoplasts was favored by alkalinization, suggesting that site Lionization influences the participation of cytochrome c in the respiratory chain and apoptosis.

1,4-Diamino-2-butanone, a wide-spectrum microbicide, yields reactive species by metal-catalyzed oxidation

The alpha-aminoketone 1,4-diamino-2-butanone (DAB), a putrescine analogue, is highly toxic to various microorganisms, including Trypanosoma cruzi. However, little is known about the molecular mechanisms underlying DAB`s cytotoxic properties. We report here that DAB (pK(a) 7.5 and 9.5) undergoes aerobic oxidation in phosphate buffer, pH 7.4, at 37 degrees C, catalyzed by Fe(II) and Cu(II) ions yielding NH(4)(+) ion, H(2)O(2), and 4-amino-2-oxobutanal (oxoDAB). OxoDAB, like methylglyoxal and other alpha-oxoaldehydes, is expected to cause protein aggregation and nucleobase lesions. Propagation of DAB oxidation by superoxide radical was confirmed by the inhibitory effect of added SOD (50 U ml(-1)) and stimulatory effect of xanthine/xanthine oxidase, a source of superoxide radical. EPR spin trapping studies with 5,5-dimethyl-1-pyrroline-1-oxide (DMPO) revealed an adduct attributable to DMPO-HO(center dot), and those with alpha-(4-pyridyl-1-oxide)-N-tert-butylnitrone or 3,5-dibromo-4-nitrosobenzenesulfonic acid, a six-line adduct assignable to a DAB(center dot) resonant enoyl radical adduct. Added horse spleen ferritin (HoSF) and bovine apo-transferrin underwent oxidative changes in tryptophan residues in the presence of 1.0-10 mM DAB. Iron release from HoSF was observed as well. Assays performed with fluorescein-encapsulated liposomes of cardiolipin and phosphatidylcholine (20:80) incubated with DAB resulted in extensive lipid peroxidation and consequent vesicle permeabilization. DAB (0-10 mM) administration to cultured LLC-MK2 epithelial cells caused a decline in cell viability, which was inhibited by preaddition of either catalase (4.5 mu M) or aminoguanidine (25 mM). Our findings support the hypothesis that DAB toxicity to several pathogenic microorganisms previously described may involve not only reported inhibition of polyamine metabolism but also DAB pro-oxidant activity. (C) 2011 Elsevier Inc. All rights reserved.

Antimicrobial Activity of Ethylenediaminedisuccinate Metal Complexes

It is shown that metal complexes of the biodegradable ligand ethylenediaminedisuccinic acid (edds) present antimicrobial activity towards fungi and bacteria. [Cd(edds)], in particular, is more toxic than free Cd2+ to Aspergillus niger, behaving as a `Trojan Horse` in the facilitated delivery of the toxic metal into the fungus.

Gruvrätten vid Stora Kopparberget 1641-1682 : en undersökning över rannsakade brott och utdömda straff

The purpose of this essay is to examine and explain how the Swedish mining court of Stora Kopparberget (the Great Copper Mountain) implemented its judicial legislation between 1641-1682. Questions are asked about which counts of indictments the court tried, which sentences they handed out, in what quantities and how these results looks in comparison with other contemporary courts. The index cards of the court judicial protocols are the primary source of information. The methods are those of quantity- and comparative analysis.The results show that theft of copper ore was the most common crime ransacked by the court. Other common crimes were (in order): sin of omission, transgression of work directions, fights, slander and disdain, trade of stolen ore, failing appearance in court etc.Fines were by far the most common sentence followed by shorter imprisonments, gauntlets, loss of right to mine possession, twig beating, loss of work, penal servitude, banishment, “wooden horse riding” and finally military transcription. Even though previous re-search, in the field of Swedish specialized courts, is almost non existent evidence confirms great similarities between the Stora Kopparberget mining court and Sala mining court. This essay will, hopefully, enrich our knowledge of specialized courts, of 17th century mining industry and society and let us reach a broader understanding of the working conditions of the mountain.

Att inte spänna kärran framför hästen : precisering av Meads ”rollövertagande”

Not to put the cart before the horse: clarification of Mead’s ”role taking” George Herbert Mead’s “role taking” is subject to many interpretations. Some dominant definitions restrict role taking to reflective people trying to understand each other and taking on each others' roles. Other definitions restrict role taking to the stage of a developed language. Against these two types of definitions I assert that Meads concept is explanatory, not merely descriptive, functionally as well as existentially. It has a value also on elementary levels of human development, where man is socially similar to other primates. Swedish sociologists have contributed fruitfully to straighten the debate. My intention is to clarify it further, and to explicate my own interpretation. Basic role taking is a non-reflective activity going on in any place where living entities influence each other with gestures in similar ways. In human beings it gives rise to conscious discursive reflectivity, and it coincides phylogenetically and ontogenetically with development of language, rather than being created by it. Role taking emanates from breaking automatic responsivity. It introduces distance and reflectivity and results in conscious social behavior, rather than being produced by such. In this way, role taking is a major step in the development of intelligence on earth.

Qu est-ce que c est?: uma HQ autobiográfica de Fábio Moon e Gabriel Bá.

Partindo da ideia de que as histórias em quadrinhos, apesar de serem consideradas um gênero híbrido, possuem também uma linguagem autônoma, este trabalho busca investigar por meio de análise de conteúdo uma história em quadrinhos (HQ)autobiográfica dos gêmeos paulistas Fábio Moon e Gabriel Bá, Qu’est-ce que c’est?, publicada pela primeira vez em inglês na coletânea Autobiographix (Dark Horse Books, 2003) e republicada em português no livro Crítica (Devir, 2004). Serão analisados recursos estéticos e narrativos, tentando descobrir também quais são as particularidades de uma HQ autobiográfica. A história narra a saga dos irmãos durante uma viagem a Paris, onde foram visitar museus e colher referências para seu trabalho. No metrô, eles se depararam com uma gangue da periferia.

Purchasing power parity: the choice of price index

Looking closely at the PPP argument, it states that the currencies purchasing power should not change when comparing the same basket goods across countries, and these goods should all be tradable. Hence, if PPP is valid at all, it should be captured by the relative price indices that best Öts these two features. We ran a horse race among six di§erent price indices available from the IMF database to see which one would yield higher PPP evidence, and, therefore, better Öt the two features. We used RER proxies measured as the ratio of export unit values, wholesale prices, value added deáators, unit labor costs, normalized unit labor costs and consumer prices, for a sample of 16 industrial countries, with quarterly data from 1975 to 2002. PPP was tested using both the ADF and the DFGLS unit root test of the RER series. The RER measured as WPI ratios was the one for which PPP evidence was found for the larger number of countries: six out of sixteen when we use DF-GLS test with demeaned series. The worst measure of all was the RER based on the ratio of foreign CPIs and domestic WPI. No evidence of PPP at all was found for this measure.

O entre do carvão ao corpo - em - arte de ator-brincante

I propose with this paper a reflection on the experiences contained in the creation of the body- in-art (FERRACINI, 2006a, b) that originated the show Rosmaninhos... This process was developed within the coletivo UZUME teatro from João Pessoa PB, through recreations and resignifications of the corporeity and physicality contained in the steps, loas, aboios, songs and choreography observed in the manners that Mestre Zequinha plays in his group of Cavalo Marinho (Sea Horse), resident in the city of Bayeux - PB, and starting from the appropriation of the text Hamlet of William Shakespeare. The body-in-art is understood in this work as a vectorial body that dilates its daily functionality, recognizing a potential learning area capable to generate creative escape lines that destabilize the "subject centered in an individuality and identity" (FOUCAULT apud FERRACINI, 2006b, p.14), being open to the differentiation of itself, indicating the possible existence of an itself-other and of the exchange-in-art space. This process of construction of the body-in-art based on Master Zequinha s ways of playing the Cavalo Marinho was methodically guided by the appropriation of the coletivo UZUME teatro of the stages of Observation, Codification and Theatricalization contained in the technique of corporal mimeses proposed by the LUME Teatro (Campinas - SP). That use resulted in two phases: Active Observation and Composition of the body-in-art. Through the repetition of these aesthetic matrixes of the Cavalo Marinho, the actors discovered actions that when, codified and organized, can configure their body-in-art, which created a vectorial exchange-in-art space to what was found in the Cavalo Marinho party. This search proposed the means of potentiating the actors' work when it comes to a preparation that allowed to dilate the scenic presence and stimulated the production of actions, which culminated in the mounting of the show Rosmaninhos...

Qualidade física e fisiológica de sementes da leguminosa forrageira Macrotyloma axillare cv. Java

É de interesse prático, quando se dispõe de diferentes lotes de sementes, conhecer a qualidade fisiológica intrínseca a cada um. Objetivou-se determinar a qualidade fisiológica de lotes de sementes da leguminosa forrageira tropical, Macrotyloma axillare cv. Java, com utilização de diferentes metodologias para realização dos testes germinação e vigor. Determinou-se a pureza física dos lotes, o peso de mil sementes, a germinação com e sem escarificação (TG) e o vigor (índice de velocidade de germinação (IVG), primeira contagem, envelhecimento acelerado e condutividade elétrica) de três lotes de sementes. Diferenças na qualidade fisiológica inicial de sementes escarificadas foram observadas pelo teste de germinação. Pelos resultados dos testes de primeira contagem e IVG não foi possível detectar diferenças na qualidade fisiológica das sementes; o envelhecimento acelerado das sementes escarificadas pode ser realizado a 41º C por 72 horas ou a 45º C por 48 horas; o teste de condutividade elétrica mostrou-se adequado para diferenciar os lotes, a partir de 48 horas de embebição.

Causes of death of thoroughbred racehorses at Octavio Dupont Veterinary Hospital, Brazilian Jockey Club, Rio de Janeiro

Existem poucas pesquisas que abordam estudos epidemiológicos de mortes em cavalos de corrida de uma forma ampla. A maioria restringe-se a uma afecção ou procedimento específico. O Brasil não possui um programa de monitoramento de mortes instituído. Através de um estudo descritivo em associação a metodologia multivariada de análise, realizou-se um estudo epidemiológico de causa mortis relacionadas com afecções musculosqueléticas (MS), gastrointestinais (GI), sistema respiratório (RES), neurológico (NEU) e mortes súbitas (SD) durante os anos de 2002 a 2008 no Hospital Octavio Dupont, Rio de Janeiro. Os machos representaram o sexo mais afetado e a causa mortis relacionada com a taxa geral de mortalidade por grandes grupos de causas determinadas apresentou a seguinte ordem decrescentemente: MS>GI>SD>NEU>RES. A maioria dos óbitos registrados foi de cavalos na faixa etária de quatro a cinco anos (ID4-ID5). Seguidamente, observaram-se as seguintes relações de correspondência: (Triênio = SM-ID>5-SD; ID>5-GI; ID4-5-MS; SF-ID<4-RES/NEU); (Quatriênio = SM - ID>5 - GI; SF - ID<4 - NEU; ID4-5 - MS; GI - ID>5). O presente estudo aponta a importância e necessidade de estudos epidemiológicos sobre lesões em cavalos atletas, objetivando o diagnostico, reconhecimento de fatores predisponentes e prevenção.

Aminophylline affects glycemia control and increases anaerobic glycolysis in horses during incremental exercise

A study was conducted on the effects of acute administration of aminophylline on physiological variables in purebred Arabian horses submitted to incremental exercise test. Twelve horses were submitted to two physical tests separated by a 10-day interval in a crossover study. These horses were divided into two groups: control (C, n = 12) and aminophylline (AM, n = 12). The drug at 10 mg/kg body weight or saline was given intravenously, 30 minutes before the incremental exercise test. The treadmill exercise test consisted of an initial warmup followed by gradually increasing physical exigency. Blood samples were assayed for lactic acid, glucose, and insulin. Maximal lactic acidemia was greater (P = .0238) in the AM group. Both V-2 and V-4 (velocities at which lactate concentrations were 2 and 4 mmol/ L, respectively) were reduced in the AM group by 15.85% (P = .0402) and 17.76% (P = .0 109), respectively. At rest as well as at 4 minutes, insulinemia was greater in the AM group (P = .0417 and .0393), Glycemia group at times 8 was statistically lower in the Al (P = .0138) and 10 minutes (P = .0432). Use of ammophylline in horses during incremental exercise does not seem to be beneficial, because this drug has a tendency to cause hypoglycemia and to increase dependence on anaerobic glucose metabolism.

Nitrogênio, fósforo, potássio, adubo orgânico e calcário dolomítico na produção de sementes de grama-batatais (Paspalum notatum Flügge) em Latossol Vermelho Escuro

Comparou-se a produção de sementes de grama-batatais em Latossol vermelho Escuro com ou sem adição de sulfato de amônio (4g de N/m²), superfosfato simples (50g de P2O5/m²), cloreto de potássio (15g de K2O/m²), esterco de equino (10 litros/m²) e calcário dolomítico (220 g/m²). As produções foram avaliadas através de número de espigas por área, peso total de sementes, peso de sementes puras, relação entre peso de sementes puras e peso total de sementes, peso de 1000 sementes puras, peso total de sementes por espiga, peso de sementes puras por espiga, porcentagem de germinação das sementes e velocidade de emergência das plântulas. Foram determinados os teores de nitrogênio, fósforo, potássio, cálcio, magnésio, enxofre, boro, cobre, ferro, manganês e zinco na matéria seca de folhas e sementes. Verificou-se que a produção de sementes, tanto em quantidade como em qualidade, foi influenciada pela nutrição mineral. As porcentagens mais elevadas de carbono no solo estiveram associadas a maior quantidade e melhor qualidade das sementes produzidas. As melhores produções de sementes correspondem aos níveis mais baixos de fósforo, potássio, cálcio e zinco nas folhas. Verificou-se serem importantes nas características avaliadas as seguintes relações entre teores de nutrientes nas folhas ou nas sementes: N/P, P/Ca, P/Mg, P/S, P/Zn, K/Ca, K/Mg, K/B, Ca/Mg, Ca/B, Fe/Mn, Zn/Cu, Zn/Fe e Zn/Mn. Nas maiores produções, as sementes apresentaram concentrações menores de nitrogênio, boro e cobre, e mais elevadas de cálcio. As sementes de melhor qualidade apresentaram maiores concentrações de zinco e menores de boro. A aplicação de adubos ou calcário dolomítico não melhorou significativamente a produção de sementes.

Determination of the highest no-effect dose (HNED) and of the elimination pattern for cocaine in horses

Cocaine is one of the most widespread illegal stimulants utilized by the human population throughout the world. The aim of this study was to establish the highest no-effect dose (HNED) of cocaine on the spontaneous locomotor activity (SLA) of horses in a behavior chamber, and thereby to determine the maximal acceptable threshold of the urinary drug concentration in horses. Twelve English thoroughbred mares received 0.02, 0.03, 0.04, 0.08 or 0.12 mg kg(-1) cocaine i.v. or saline solution (control). It was noted that doses above 0.04 mg kg(-1) induced a significant increase in SLA (P < 0.05, Tukey's test). No significant increase in SLA was seen in the mares that received 0.03 mg kg(-1), but the animals showed important behavioral changes that did not occur after the 0.02 mg kg(-1) dose. It was concluded that the HNED of cocaine for horses in a behavior chamber is 0.02 mg kg(-1). After injection of this dose in five horses, urine samples were collected at predetermined intervals through vesical catheterization. The concentrations of cocaine, norcocaine, benzoylecgonine and ecgonine methyl ester were quantified by liquid chromatography/electrospray ionization tandem mass spectrometry. Cocaine and norcocaine concentrations remained consistently below the level of detection. Benzoylecgonine reached a mean (+/- SEM) maximum concentration of 531.9 +/- 168.7 ng ml(-1) after 4 h, whereas ecgonine methyl ester peaked 2 h after injection at a concentration of 97.2 +/- 26.5 ng ml(-1). The maximum admissible concentration for cocaine and/or metabolites in the urine of horses is difficult to establish unequivocally because of the substantial individual variation in the drug elimination pattern observed in horses, which can be inferred by the large standard error of the means obtained. Copyright (C) 2002 John Wiley Sons, Ltd.

Hemogasometria de eqüinos submetidos à obstrução experimental do duodeno, íleo e cólon maior

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

Efeitos comportamental, clínico e analgésico promovidos pela injeção epidural preventiva de morfina, xilazina ou clonidina, em equinos

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