Preimplantation development and expression of Hsp-70 and Bax genes in bovine blastocysts derived from oocytes matured in alpha-MEM supplemented with growth factors and synthetic macromolecules

In vitro culture conditions affect both the maternal and embryonic expression of genes and is likely to alter both oocyte and embryo developmental competence. The search for better and less variable culture conditions simulating those in vivo has led to the development of defined culture media, with lower impact on the molecular reprogramming of oocytes and embryos. We evaluated embryo development and relative abundance (RA) of Hsp-70 and Bax transcripts in bovine blastocysts produced from oocytes matured in a chemically defined IVM system with synthetic polymers. Immature cumulus oocyte complexes (COCs) were matured for 22-24 h in alpha-MEM supplemented with IGF-1, insulin, 0.1% polyvinyl alcohol (PVA), or 0.1% polyvinylpyrrolidone (PVP), but without FSH or LH. The control group consisted of COCs matured it, TCM plus FSH and 10% estrous cow serum. After fertilization. presumptive zygotes were co-cultured with cumulus cells until 224 h post-insemination. Total RNA was isolated from embryo pools, reverse transcribed into cDNA, and subjected to transcript analysis by real-time PCR. Cleavage rate was higher (P < 0.05) for the control group (68.3%) than for the PVA (54.4%) and PVP-40 (58.3%) groups. Nevertheless. there was no difference among the PVA, PVP-40 and control groups in blastocyst or hatching rates. similarly, no difference in relative abundance of Hsp-70 and Bax transcripts was detected in comparison to the control group. We inferred that bovine oocytes can be matured in serum- and gonadotrophin-free medium supplemented with PVA or PVP, enriched with IGF-I and insulin, without altering post-cleavage development and relative abundance of some genes associated with stress and apoptosis. (C) 2009 Elsevier Inc. All rights reserved.

Estudo eletroquímico comparativo do aço ISO 5832-9 em diferentes meios de interesse biológico

Foi estudado o comportamento eletroquímico a 37°C do aço inoxidável ISO 5832-9, em meios de NaCl 0,9 %, de Ringer Lactato e meio mínimo de Eagle (MEM), por voltametria linear e análises da superfície por microscopia eletrônica de varredura (MEV) e por espectroscopia por dispersão de energia (EDS). Foram feitos ensaios mecânicos e testes de toxicidade. O aço ISO 5832-9 se encontra passivado no potencial de corrosão e não apresenta corrosão por pite nos três meios estudados em toda faixa de potencial investigada, desde o potencial de corrosão até 50 mV acima do potencial de transpassivação. Em meio de MEM, no entanto, as análises por MEV e EDS mostraram que o referido aço, nesse valor mais elevado de potencial, apresentou um comportamento diferente, com perda das inclusões de óxido de manganês. Os potenciais de corrosão, Ecorr (potencial de circuito aberto estacionário) bem como os valores de densidade de corrente de passivação, variaram na seguinte ordem: Ecorr, RL < Ecorr, NaCl < Ecorr, MEM. e jMEM << jRL ≅ jNaCl. No ensaio de citotoxicidade, o aço foi caracterizado como não-tóxico.

Análise espacial e sazonal da leptospirose no município de São Paulo, SP, 1998 a 2006

OBJETIVO: Analisar a distribuição espacial e sazonal da leptospirose, identificando possíveis componentes ecológicos e sociais para a sua transmissão. MÉTODOS: Foram georreferenciados 2.490 casos em cada distrito do município de São Paulo, SP, registrados de 1998 a 2006. Os dados foram obtidos do Sistema de Informação de Agravos de Notificação. Foram realizados mapas temáticos com as variáveis taxa de incidência, letalidade, taxa de alfabetização, renda média mensal, número de moradores por domicilio, abastecimento de água e rede de esgoto. Para identificar o padrão espacial (disperso, em aglomerado ou randômico), foram analisadas pelo Índice de Moran global e local. Foi utilizado o coeficiente de correlação de Spearman para testar associações entre as variáveis com padrão espacial em aglomerados. RESULTADOS: O padrão espacial em aglomerados foi observado nas variáveis taxa de incidência de leptospirose, taxa de alfabetização, renda média mensal, número de moradores por domicílio, abastecimento de água e rede de esgoto. Foram notificados 773 casos no período seco e 1.717 no úmido. A incidência e a letalidade estão correlacionadas com as condições socioeconômicas da população, independentemente do período. CONCLUSÕES: A leptospirose está distribuída por todo o município de São Paulo e sua incidência aumenta no período das chuvas. No período seco, os locais de aparecimento dos casos coincidem com as áreas de piores condições de moradia e, durante o período úmido, também aumenta em outros distritos, provavelmente devido à proximidade de rios e córregos.

Memantine prevents cardiomyocytes nuclear size reduction in the left ventricle of rats exposed to cold stress

OBJECTIVES: Memantine is an N-methyl-d-aspartate (NMDA) glutamate receptor antagonist used to treat Alzheimer's disease. Previous studies have suggested that receptor blockers act as neuroprotective agents; however, no study has specifically investigated the impact that these drugs have on the heart. We sought to evaluate the effects of memantine on nuclear size reduction in cardiac cells exposed to cold stress. METHOD: We used male EPM-Wistar rats (n=40) divided into 4 groups: 1) Matched control (CON); 2) Memantine-treated rats (MEM); 3) Rats undergoing induced hypothermia (IH) and 4) Rats undergoing induced hypothermia that were also treated with memantine (IHM). Animals in the MEM and IHM groups were treated by oral gavage administration of 20 mg/kg/day memantine over an eight-day period. Animals in the IH and IHM groups were submitted to 4 hours of hypothermia in a controlled environment with a temperature of - 8ºC on the last day of the study. RESULTS: The MEM group had the largest cardiomyocyte nuclear size (151 ± 3.5 μm³ vs. CON: 142 ± 2.3 μm³; p<0.05), while the IH group had the smallest mean value of nuclear size. The nuclear size of the IHM group was preserved (125 ± 2.9 μm³) compared to the IH group (108 ± 1.7 μm³; p<0.05). CONCLUSION: Memantine prevented the nuclear size reduction of cardiomyocytes in rats exposed to cold stress.

Continuous monitoring of ascorbate transport through neuroblastoma cells with a ruthenium oxide hexacyanoferrate modified microelectrode

The uptake of ascorbate by neuroblastoma cells using a ruthenium oxide hexacyanoferrate (RuOHCF)-modified carbon fiber disc (CFD) microelectrode (r = 14.5 mu m) was investigated. By use of the proposed electrochemical sensor the amperometric determination of ascorbate was performed at 0.0 V in minimum essential medium (MEM, pH = 7.2) with a limit of detection of 25 mu mol L(-1). Under the optimum experimental conditions, no interference from MEM constituents and reduced glutathione (used to prevent the oxidation of ascorbate during the experiments) was noticed. The stability of the RuOHCF-modified electrode response was studied by measuring the sensitivity over an extended period of time (120 h), a decrease of around 10% being noticed at the end of the experiment. The rate of ascorbate uptake by control human neuroblastoma SH-SY5Y cells, and cells transfected with wild-type Cu,Zn-superoxide dismutase (SOD WT) or with a mutant typical of familial amyotrophic lateral sclerosis (SOD G93A), was in agreement with the level of oxidative stress in these cells. The usefulness of the RuOHCF-modified microelectrode for in vivo monitoring of ascorbate inside neuroblastoma cells was also demonstrated.

Política externa chilena e espectro ideológico político-partidário: um estudo sobre a Câmara dos Deputados (2002-2006)

The influence of political parties on decisions made by members of Congress is a hotly debated issue in political science. In foreign policy, which is usually considered nonpartisan, the matter is even more inconclusive. The current study analyzes all the roll-call votes taken on foreign policy issues in the 2002-2006 legislature of the Chilean Chamber of Deputies. After tracing a spatial map of foreign policy preferences among Chilean Deputies using the Nominate statistical package, we concluded that the ideology of the legislator's political party is a predictive factor for his or her foreign policy behavior. Our findings indicate that the way Chilean legislators structure their preferences on foreign policy issues does not differ significantly from the way they shape their domestic policy preferences.

The electrochemical behaviour of Ti-13Nb-13Zr alloy in various solutions

The electrochemical behaviour of a near-beta Ti-13Nb-13Zr alloy for the application as implants was investigated in various solutions. The electrolytes used were 0.9 wt% NaCl solution, Hanks` solution and a culture medium known as minimum essential medium (MEM) composed of salts, vitamins and amino acids, all at 37 degrees C. The electrochemical behaviour was investigated by the following electrochemical techniques: open circuit potential measurements as a function of time, electrochemical impedance spectroscopy (EIS) and determination of polarisation curves. The obtained results showed that the Ti alloy was passive in all electrolytes. The EIS results were analysed using an equivalent electrical circuit representing a duplex structure oxide layer, composed of an inner barrier layer, mainly responsible for the alloy corrosion resistance, and an outer and porous layer that has been associated to osteointegration ability. The properties of both layers were dependent on the electrolyte used. The results suggested that the thickest porous layer is formed in the MEM solution whereas the impedance of the barrier layer formed in this solution was the lowest among the electrolytes used. The polarisation curves showed a current increase at potentials around 1300 mV versus saturated calomel electrode (SCE), and this increase was also dependent on the electrolyte used. The highest increase in current density was also associated to the MEM solution suggesting that this is the most aggressive electrolyte to the Ti alloy among the three tested solutions.

Spatial genetic structure of Hymenaea stigonocarpa Mart. ex Hayne assessed with chloroplast microsatellite markers

The use of chloroplast DNA markers (cpDNA) helps to elucidate questions related to ecology, evolution and genetic structure. The knowledge of inter-and intra-population genetic structure allows to design effective conservation and management strategies for tropical tree species. With the aim to help the conservation of Hymenaea stigonocarpa of the Cerrado (Brazilian savanna) in Sao Paulo State, an analysis of the spatial genetic structure (SGS) was conducted in two populations using five universal chloroplast microsatellite loci (cpSSR). The population of 68 trees of H. stigonocarpa in the Ecological Station of Itirapina (ESI) had a single haplotype, indicating a strong founder effect. In turn, the population of 47 trees of H. stigonocarpa in a contiguous area that includes the Ecological Station of Assis and the Assis State Forest (ESA), showed six haplotypes ((n) over cap (h) = 6) with a moderate haplotype diversity ((h) over cap = 0667 + 0094), revealing that it was founded by a small number of maternal lineages. The SGS analysis for the population ESA/ASF, using Moran`s I index, indicated limited seed dispersal. Considering SGS, for ex situ conservation strategies in the population ESA/ASF, seed harvesting should require a minimum distance of 750 m among seed-trees.

CONGENITAL CLUBFOOT

The clubfoot is one of the most common congenital deformities affecting the lower limbs, it still presents controversial aspects regarding etiology and treatment. In spite of its relatively high frequency, the treatment is still challenging, since the long-term aim is achieving an everlasting flexible, plantigrade, pain-free and totally functional foot. The Ponseti method has gained attention and popularity because of its satisfactory results and surgery avoidance. Presently, surgical treatment is indicated only after failure of conservative methods, avoiding extensive soft-tissue release, but performing localized corrections of the deformities, a technique also know as ""a la carte"" release. The future perspective is based on the knowledge about long-term results and new understanding of the clubfoot etiology, especially in the genetic field, which may eventually be helpful for prognostic and treatment. Level of Evidence: Level II, systematic review.