Genomic characterization of oxacillin-resistant Staphylococcus epidermidis and Staphylococcus haemolyticus isolated from Brazilian medical centres.

Studies on the genetic diversity of oxacillin-resistant coagulase-negative staphylococcal (CNS) isolates are important for the control and prevention of infections. The present study evaluated the clonal diversity of oxacillin-resistant Staphylococcus epidermidis (ORSE) and Staphylococcus haemolyticus (ORSH) strains, isolated from patients in nine Brazilian medical centres by using pulsed-field gel electrophoresis (PFGE) after digestion of bacterial DNA using SmaI. PFGE analysis of ORSE (N=44) and ORSH (N=25) strains showed the presence of 29 restriction profiles clustered in 16 PFGE types, and 21 distinct profiles in 15 PFGE types, respectively, indicating a large genetic diversity among isolates of both of these species. Among the ORSE isolates, 23 (52%) strains belonged to two predominant PFGE types (named A and B), which were observed in most of the hospitals assessed, indicating the spread of these PFGE types in hospitals located in Rio de Janeiro. The spread of PFGE types of ORSH was also detected in some of the hospitals investigated. The results show that PFGE is a suitable tool for epidemiological studies of oxacillin-resistant CNS, and can be used as a basis for infection control procedures for these multiresistant organisms.

Calcium- and magnesium-dependent aggregation of legume seed storage proteins.

The solubility characteristics and sedimentation behavior of total or individual globulins from legume seeds [Lupinus albus L., Pisum sativum L., and Glycine max (L.) Merr.] were investigated. The typical insolubility of globulins detected during their extraction seems to be due to the presence of a low molecular weight factor(s) in the seed extract. The solubility of the purified globulins decreases with increasing concentrations of calcium and/or magnesium, but not of other cations, showing minimum values at concentrations that vary with the particular globulin considered. Ultracentrifugation analyses revealed that the Ca(2+)- and/or Mg(2+)-induced insolubilization of the globulins involves the formation of high-order aggregates of molecules of the same or of different globulins. These macromolecular structures are dissociated under conditions of high ionic strength, suggesting the involvement of electrostatic interactions in the aggregation process. The degree of association relies heavily on the amount of Ca(2+) and/or Mg(2+) available, on the presence of chelating agents for these divalent cations, and on the ionic strength of the surrounding medium. The possible physiological significance of the findings is discussed.

Storage proteins from Lathyrus sativus seeds.

The proteins from Lathyrus sativus Linn. (chickling vetch or grass pea) seeds were investigated. Protein constitutes approximately 20% of the seed dry weight, >60% of which is composed by globulins and 30% by albumins. A single, 24 kDa polypeptide comprises more than half of the protein present in the albumin fraction. The globulins may be fractionated into three main components, which were named alpha-lathyrin (the major globulin), beta-lathyrin, and gamma-lathyrin. alpha-Lathyrin, with a sedimentation coefficient of approximately 18S, is composed of three main types of unglycosylated subunits (50-66 kDa), each of which produce, upon reduction, a heavy and a light polypeptide chain, by analogy with 11S. beta-Lathyrin, with a sedimentation coefficient of 13S, is composed by a relatively large number of subunits (8-66 kDa). Two major polypeptides are glycosylated and exhibit structural similarity with beta-conglutin from Lupinus albus. One of these possesses an internal disulfide bond. gamma-Lathyrin, with a sedimentation coefficient of approximately 5S, contains two interacting, unglycosylated polypeptides, with no disulfide bonds: the major 24 kDa albumin and the heavier (20 kDa) polypeptide chain of La. sativus lectin.

The wide diversity of structurally similar wine proteins.

In the present work, single grape variety wines, Moscatel and Arinto, were used. Analysis by denaturing polyacrylamide gel electrophoresis of the wine proteins revealed the presence of only a few polypeptides ranging in molecular mass from 15 to 30 kDa. However, a more detailed examination of the whole protein fraction, by a combination of techniques, showed that these wines contain a very large number (many tens and, possibly, many more) of distinct polypeptides, exhibiting similar molecular masses but different electrical charges. The results obtained using highly specific antibodies and N-terminal sequencing indicate that there is structural similarity among most of the wine polypeptides. These observations can be explained by the existence of a common precursor to most or all of the wine proteins, which could generate all of the detected polypeptides by limited proteolysis. Comparison of the N-terminal sequences of the polypeptides isolated from Moscatel wine with proteins from other sources revealed a high degree of homology to pathogenesis-related proteins.

Implementation of a Brazilian Cardioprotective Nutritional (BALANCE) Program for improvement on quality of diet and secondary prevention of cardiovascular events: A randomized, multicenter trial

Background Complex percutaneous coronary intervention (PCI) is associated with higher ischemic risk, which can be mitigated by long-term dual antiplatelet therapy (DAPT). However, concomitant high bleeding risk (HBR) may be present, making it unclear whether short- or long-term DAPT should be prioritized. Objectives This study investigated the effects of ischemic (by PCI complexity) and bleeding (by PRECISE-DAPT [PRE dicting bleeding Complications in patients undergoing stent Implantation and Sub sequent Dual Anti Platelet Therapy] score) risks on clinical outcomes and on the impact of DAPT duration after coronary stenting. Methods Complex PCI was defined as ≥3 stents implanted and/or ≥3 lesions treated, bifurcation stenting and/or stent length >60 mm, and/or chronic total occlusion revascularization. Ischemic and bleeding outcomes in high (≥25) or non-high (<25) PRECISE-DAPT strata were evaluated based on randomly allocated duration of DAPT. Results Among 14,963 patients from 8 randomized trials, 3,118 underwent complex PCI and experienced a higher rate of ischemic, but not bleeding, events. Long-term DAPT in non-HBR patients reduced ischemic events in both complex (absolute risk difference: −3.86%; 95% confidence interval: −7.71 to +0.06) and noncomplex PCI strata (absolute risk difference: −1.14%; 95% confidence interval: −2.26 to −0.02), but not among HBR patients, regardless of complex PCI features. The bleeding risk according to the Thrombolysis In Myocardial Infarction scale was increased by long-term DAPT only in HBR patients, regardless of PCI complexity. Conclusions Patients who underwent complex PCI had a higher risk of ischemic events, but benefitted from long-term DAPT only if HBR features were not present. These data suggested that when concordant, bleeding, more than ischemic risk, should inform decision-making on the duration of DAPT. (AU)

Acompanhamento eletrocardiográfico no desenvolvimento de ratos Wistar / Eletrocardiographic monitoring in the development of Wistar Rats

A atividade elétrica cardíaca pode ser registrada por meio do eletrocardiograma (ECG), registro capaz de captar as alterações do percurso elétrico. Na literatura científica, não existe descrição de padrões eletrocardiográficos de ratos Wistar desde a fase de recém-nascidos até serem considerados envelhecidos. O objetivo do presente estudo foi avaliar o padrão eletrocardiográfico de ratos Wistar desde a fase de recém-nascidos até serem considerados envelhecidos, no intuito de que os resultados possam contribuir como referencial dos padrões da espécie. Foram utilizados ratos Rattus novergicus albinus, da linhagem Wistar. Os ratos foram divididos em cinco grupos experimentais, sendo definidos de acordo com a idade e assim denominados: grupo 21 dias (21D), dois meses (2M), três meses (3M), sete meses (7M) e 18 meses (18M), sendo n=20 por grupo. Para a realização do ECG, eletrodos conectados aos canais do eletrocardiógrafo foram posicionados nos animais e captaram o sinal elétrico do coração em seis derivações. Os dados foram agrupados e submetidos a comparação por meio de ANOVA e teste de Tukey-Kramer, P≤0,05. Os resultados mostraram que, de forma geral, as modulações ao longo do processo de envelhecimento diante dos parâmetros relacionados ao ECG se iniciam a partir do período 2M, com aparente estabilidade até 3M e/ou 7M, seguindo por exacerbação das modulações em 7M e/ou 18M, a depender da variável eletrocardiográfica em questão. Em relação à frequência cardíaca (FC), concomitantemente, nota-se um padrão similar, ou seja, diminuição dela a partir do período 2M, com estabilidade até 7M, seguindo por diminuição mais pronunciada em 18M. Os resultados sugerem que, devido ao processo de envelhecimento, o coração de ratos Wistar apresenta modificações na condução do estímulo elétrico, bem como redução na FC, fatores que merecem atenção enquanto indicadores de inclusão/exclusão de animais em grupos experimentais por parte de [...].(AU)

The cardiac electrical activity can be recorded through the electrocardiogram (ECG), this recording is able to capture the changes in the electric route. In the scientific literature, there is no description of electrocardiographic patterns of Wistar rats at the stage of newborn to be considered aged. The aim of this study was to evaluate the electrocardiographic pattern of Wistar rats at various stages of growth, in order for the results to contribute as a standard reference. Rattus norvegicus albinus rats of the Wistar lineage were used. The rats were divided into 5 experimental groups being defined according to age and named as follows: group 21 days (21D), 2 months (2M), 3 months (3M), 7 months (7M) and 18 months (18M), where n = 20 per group. For the realization of ECG, electrodes connected to the electrocardiograph channels were positioned in animals to capture the electrical signal of the heart in 6-lead. The data were grouped and subjected to comparison by ANOVA and Tukey-Kramer test, P=0.05. The results showed that, overall, the modulations along the front aging process, related to the ECG, are initiated from 2M period, apparent stability by 3M and/or 7M, followed by exacerbation of modulations in 7M and/or 18M, depending on the electrocardiographic variable in question. Front of heart rate (HR), concomitantly, there is a similar pattern, that is, decreasing from 2M period with stability until 7M, followed by more pronounced decrease in 18M. The results suggest that due to the aging process, the heart of Wistar rats shows changes in the conduction of electrical stimulation and reduction in heart rate, factors that deserve attention as indicators of inclusion / exclusion of animals in experimental groups by researchers using that species as a model.(AU)

Acompanhamento eletrocardiográfico no desenvolvimento de ratos Wistar / Eletrocardiographic monitoring in the development of Wistar Rats

A atividade elétrica cardíaca pode ser registrada por meio do eletrocardiograma (ECG), registro capaz de captar as alterações do percurso elétrico. Na literatura científica, não existe descrição de padrões eletrocardiográficos de ratos Wistar desde a fase de recém-nascidos até serem considerados envelhecidos. O objetivo do presente estudo foi avaliar o padrão eletrocardiográfico de ratos Wistar desde a fase de recém-nascidos até serem considerados envelhecidos, no intuito de que os resultados possam contribuir como referencial dos padrões da espécie. Foram utilizados ratos Rattus novergicus albinus, da linhagem Wistar. Os ratos foram divididos em cinco grupos experimentais, sendo definidos de acordo com a idade e assim denominados: grupo 21 dias (21D), dois meses (2M), três meses (3M), sete meses (7M) e 18 meses (18M), sendo n=20 por grupo. Para a realização do ECG, eletrodos conectados aos canais do eletrocardiógrafo foram posicionados nos animais e captaram o sinal elétrico do coração em seis derivações. Os dados foram agrupados e submetidos a comparação por meio de ANOVA e teste de Tukey-Kramer, P≤0,05. Os resultados mostraram que, de forma geral, as modulações ao longo do processo de envelhecimento diante dos parâmetros relacionados ao ECG se iniciam a partir do período 2M, com aparente estabilidade até 3M e/ou 7M, seguindo por exacerbação das modulações em 7M e/ou 18M, a depender da variável eletrocardiográfica em questão. Em relação à frequência cardíaca (FC), concomitantemente, nota-se um padrão similar, ou seja, diminuição dela a partir do período 2M, com estabilidade até 7M, seguindo por diminuição mais pronunciada em 18M. Os resultados sugerem que, devido ao processo de envelhecimento, o coração de ratos Wistar apresenta modificações na condução do estímulo elétrico, bem como redução na FC, fatores que merecem atenção enquanto indicadores de inclusão/exclusão de animais em grupos experimentais por parte de pesquisadores que utilizam a referida espécie como modelo de estudo.(AU)

The cardiac electrical activity can be recorded through the electrocardiogram (ECG), this recording is able to capture the changes in the electric route. In the scientific literature, there is no description of electrocardiographic patterns of Wistar rats at the stage of newborn to be considered aged. The aim of this study was to evaluate the electrocardiographic pattern of Wistar rats at various stages of growth, in order for the results to contribute as a standard reference. Rattus norvegicus albinus rats of the Wistar lineage were used. The rats were divided into 5 experimental groups being defined according to age and named as follows: group 21 days (21D), 2 months (2M), 3 months (3M), 7 months (7M) and 18 months (18M), where n = 20 per group. For the realization of ECG, electrodes connected to the electrocardiograph channels were positioned in animals to capture the electrical signal of the heart in 6-lead. The data were grouped and subjected to comparison by ANOVA and Tukey-Kramer test, P=0.05. The results showed that, overall, the modulations along the front aging process, related to the ECG, are initiated from 2M period, apparent stability by 3M and/or 7M, followed by exacerbation of modulations in 7M and/or 18M, depending on the electrocardiographic variable in question. Front of heart rate (HR), concomitantly, there is a similar pattern, that is, decreasing from 2M period with stability until 7M, followed by more pronounced decrease in 18M. The results suggest that due to the aging process, the heart of Wistar rats shows changes in the conduction of electrical stimulation and reduction in heart rate, factors that deserve attention as indicators of inclusion / exclusion of animals in experimental groups by researchers using that species as a model.(AU)

Protein Degradation in Lemna with Particular Reference to Ribulose Bisphosphate Carboxylase: II. The Effect of Nutrient Starvation.

The concept of ribulose bisphosphate carboxylase as a storage protein is not supported in the case of Lemna minor, where the enzyme appears to be particularly stable under conditions of nitrogen starvation. Total nutrient starvation in light and in the dark induced the degradation of this enzyme, but not at an enhanced rate compared with other leaf proteins and, surprisingly, darkness inhibited the degradation of chlorophyll which occurs with total nutrient starvation in the light. The data suggest that Lemna is not programmed to senesce in response to nutrient starvation. Differences in the pattern of protein degradation, which occurred under the stress conditions employed, are not consistent with a simple model of protein degradation in which the degradative system is assumed to be located in the vacuole. The data is best explained by a dual system in which cytosolic proteins are degraded by a vacuolar/lysosomal system and chloroplast proteins are degraded within the chloroplast. Whatever the system of degradation, our data do not support the proposed correlation between the rate of protein degradation and either protein charge or size.

Protein degradation in lemna with particular reference to ribulose bisphosphate carboxylase: I. The effect of light and dark.

Ribulose bisphosphate carboxylase from Lemna minor resembles the structure reported for the enzyme from other plants. When grown in the light, the enzyme appears to undergo little or no degradation, as measured by a double-isotope method. This situation is similar to that reported for wheat and barley, but is unlike that reported for maize, where the enzyme degrades at the same rate as total protein. Prolonged periods of darkness usually induce leaf senescence, characterized by the rapid degradation of chlorophyll and protein, with ribulose bisphosphate carboxylase undergoing preferential degradation. In L. minor there is selective protein degradation in the dark, but chlorophyll and ribulose bisphosphate carboxylase are stable when fronds are kept in the darkness for up to 8 days. It appears that Lemna is not programmed to senesce, or at least that darkness does not induce senescence in Lemna. Although there is no evidence for in vivo degradation or modification of ribulose bisphosphate carboxylase during prolonged periods of darkness, extracts from fronds which have been kept in the dark for periods in excess of 24 hours convert ribulose bisphosphate carboxylase to a more acidic form. The properties of the dark-induced system which acts on ribulose bisphosphate carboxylase, suggest that it may be a mixed function oxidase. The proposition that the selectivity of protein degradation is genetically determined, so that the rate at which a protein is degraded is determined by its charge or size, was tested for fronds grown in the light or maintained in the dark. There was no significant correlation between protein degradation and either charge or size, in light or dark.

Is protein degradation correlated with either the charge or size of Lemna proteins?

Evidence is presented that the organelles of Lemna minor do not degrade as functional units. The proteins of Lemna show wide differences in their rates of degradation and ribulose bisphosphate carboxylase (EC 4.1.1.39) has a particularly slow rate of degradation. Contrary to some earlier evidence, we found no correlation between the rate of soluble-protein degradation and either charge or size of proteins. We could find no correlation between protein degradation and subunit size in any of the organelles of Lemna.

Conversion of ribulose-1,5-bisphosphate carboxylase to an acidic and catalytically inactive form by extracts of osmotically stressed Lemna minor fronds.

The fronds of Lemna minor L. respond to a number of stresses, and in particular to an osmotic stress, by producing an enzyme system which catalyzes the oxidation of ribulose-1,5-bisphosphate carboxylase (RuBPCase; EC 4.1.1.39) to an acidic and catalytically inactive form. During the first 24 h of osmotic stress the induced oxidase system does not seem to exert a significant in-vivo effect on RuBPCase, presumably because of compartmentation. Subsequently, the oxidase system gains access to the enzyme and converts it to the acid and catalytically inactive form and eventually the oxidase system declines in activity.A number of partially acidified forms of RuBPCase are formed during oxidation, and this process appears to be correlated with the disappearance of varying numbers of SH residues. The number of-SH residues in RuBPCase from Lemna has been estimated at 89. However, RuBPCase isolated from 24-h osmotically stressed fronds showed a reduction in the number of-SH residues per molecule from 89 to 54. It seems likely that the oxidation of-SH groups is causally related to the acidification of RuBPCase which occurs during osmotic stress.

Effect of osmotic stress on protein turnover in Lemna minor fronds.

Evidence is presented that although many proteins from the fronds of Lemna minor L. undergo enhanced degradation during osmotic stress, ribulose-1,5-bisphosphate carboxylase (RuBPCase) is not degraded. Instead RuBPCase is converted in a series of steps to a very high-molecular-weight form. The first step involves the induction of an oxidase system which after 24 h of stress converts RuBPCase to an acidic and catalytically inactive form. Subsequently, the oxidised RuBPCase protein is gradually polymerized to a number of very large aggregates (molecular weight of several million).The conversion of RuBPCase to a high-molecular-weight form appears to be correlated with (i) a reduction in the number of-SH residues and (ii) the susceptibility to in-vitro proteolysis. Indeed, the number of-SH groups per RuBPCase molecule decreases from 89 in the native enzyme to 54 and 22 in the oxidised and polymerized forms, respectively. On the other hand, the oxidised enzyme is more susceptible to in-vitro proteolysis than the native form. However, it is the polymerized form of RuBPCase which is particularly susceptible to in-vitro proteolysis.Western-blotting experiments and anti-ubiquitin antibodies were used to detect the presence of ubiquitin conjugates in extracts from osmotically stressed Lemna fronds. The possible involvement of ubiquitin in the formation of the aggregates is discussed.

Use of a single method in the extraction of the seed storage globulins from several legume species. Application to analyse structural comparisons within the major classes of globulins.

In this study, a single, improved methodology was used to extract, fractionate and purify the 11S (legumin-type or related to the alpha-conglutin from Lupinus albus L.), 7S (vicilin-type or related to the beta-conglutin from L. albus) and 2S (related to the gamma-conglutin from L. albus) families of proteins from eight legume species: L. albus, Glycine max (L.) Merr., Pisum sativum L., Vicia faba L., Cicer arietinum L., Phaseolus vulgaris L., Lens culinaris Med. and Arachis hypogaea L. The sedimentation coefficients obtained varied from 1.9 to 8.1 for the gamma-conglutin-related proteins, from 5.1 to 10.5 for the beta-conglutin-related proteins and from 12.0 to 14.9 for the alpha-conglutin-related globulins. The gamma-conglutin-related proteins is the most heterogeneous group. Antibodies produced against each type of gamma-conglutin polypeptide chain recognize the other polypeptide chain as well as other polypeptides in the corresponding globulins from all species examined. The 7S globulins are typically composed of a large number of polypeptides, covering a wide range of molecular masses (10 to 70 kD). The presence of disulphide bonds is apparently absent and the occurrence of glycopolypeptides is not widespread. Finally, the 11S globulins are characteristically formed by a limited number of polypeptides that may be divided into a lighter group (20-25 kD) and a heavier group (35-50 kD). The presence of disulphide bonds is apparently widespread but the occurrence of glycopolypeptides seems to be relatively rare. Both the 7S family and the 11S globulins studied by immunoblotting exhibit a low level of structural similarity.

Simulação computacional de estratégias de manutenção de equipamentos de mamografia / Computer simulation of maintenance strategies of mammography equipment

The elaboration of a maintenance strategy is a main concern for hospital administrators. The strategies can be optimized through computational simulation of discrete events. The objective of this work is to present a model of a standard mammography clinic and to analyze the impact of different mammograph maintenance strategies on the clinic routine...

Análise de desempenho do fluxo de atendimento hospitalar com auxílio da simulação computacional / Performance analysis of the flow of patient care with the aid of computer simulation

Healthcare management is a dificult enterprise, which can be helped by modern computacional approaches such as the use of models based on computer simulation. The objective of this studt is to present an application of this tool to the analysis of the performance of a hospital clinic (an ophthalmologic clinic in a large hospital)...