Protection of nonobese diabetic mice from autoimmune diabetes by reduction of islet mass before insulitis.

Nonobese diabetic mice spontaneously develop diabetes that is caused by autoimmune cell-mediated destruction of pancreatic beta cells. Here we report that surgical removal of 90% of pancreatic tissue before onset of insulitis induced a long-term diabetes-free condition in nonobese diabetic mice. Pancreatectomy after development of moderate insulitis had no effect on the course of diabetes. The effect of pancreatectomy was abrogated with subsequent development of diabetes by infusion of islet-cell-specific T lymphocytes and by transplantation of pancreatic islets. Lymphocytes from pancreatectomized diabetes-free mice exhibited low response to islet cells but responded normally to alloantigens. These results suggest that the islet cell mass plays a critical role in development of autoimmune diabetes.

Probing the oligomeric structure of an enzyme by electrospray ionization time-of-flight mass spectrometry.

Electrospray ionization time-of-flight (ESI-TOF) mass spectrometry was used to study the quaternary structure of 4-oxalocrotonate tautomerase (EC 5.3.2; 4OT), and four analogues prepared by total chemical synthesis. Wild-type 4OT is a hexamer of 62 amino acid subunits and contains no cysteine residues. The analogues were: (desPro1)4OT, a truncated construct in which Pro1 was deleted; (Cpc1)4OT in which Pro1 was replaced with cyclopentane carboxylate; a derivative [Met(O)45]4OT in which Met45 was oxidized to the sulfoxide; and an analogue (Nle45)4OT in which Met45 was replaced with norleucine. ESI of (Nle45)4OT, (Cpc1)4OT, and 4OT from solution conditions under which the native enzyme was fully active (5 mM ammonium bicarbonate buffer, pH 7.5) gave the intact hexamer as the major species detected by TOF mass spectrometry. In contrast, analysis of [Met(O)45]4OT and (desPro1)4OT under similar conditions yielded predominantly monomer ions. The ESI-TOF measurements were consistent with structural data obtained from circular dichroism spectroscopy. In the context of kinetic data collected for 4OT and these analogues, ESI-TOF mass spectrometry also provided important evidence for the structural and mechanistic significance of the catalytically important Pro1 residue in 4OT.

DNA sequencing by delayed extraction-matrix-assisted laser desorption/ionization time of flight mass spectrometry.

Matrix-assisted laser desorption/ionization (MALDI) time of flight mass spectrometry was used to detect and order DNA fragments generated by Sanger dideoxy cycle sequencing. This was accomplished by improving the sensitivity and resolution of the MALDI method using a delayed ion extraction technique (DE-MALDI). The cycle sequencing chemistry was optimized to produce as much as 100 fmol of each specific dideoxy terminated fragment, generated from extension of a 13-base primer annealed on 40- and 50-base templates. Analysis of the resultant sequencing mixture by DE-MALDI identified the appropriate termination products. The technique provides a new non-gel-based method to sequence DNA which may ultimately have considerable speed advantages over traditional methodologies.

Programa Bolsa Família na coorte de nascimentos de Pelotas, 2004: focalização e impactos no IMC/Idade, obesidade e composição corporal de crianças

Introdução: O Programa Bolsa Família é a principal estratégia brasileira para amenizar a pobreza e vulnerabilidade social, com diferentes impactos na vida dos beneficiários. O aumento da renda, em função do benefício, poderia trazer resultados positivos na alimentação, uma vez que possibilitam uma maior diversidade da dieta. Porém, poderia trazer resultados negativos como a ingestão excessiva de energia e consequente aumento da adiposidade. As avaliações dos impactos do programa em termos de obesidade e massa gorda de crianças são inexistentes. Objetivo: Avaliar o impacto do Programa Bolsa Família no estado nutricional (IMC/idade) e na composição corporal aos 6 anos de idade entre as crianças da Coorte de Nascimentos de Pelotas (RS), 2004. Métodos: Os dados foram provenientes da integração dos bancos da Coorte de Nascimentos de Pelotas de 2004 e do Cadastro Único do Governo Federal. Foi realizada análise descritiva da cobertura e focalização do programa, com informações do nascimento e dos 6 anos de idade (n=4231). Considerou-se focalização o percentual de elegíveis entre o total de beneficiários e cobertura o percentual de famílias elegíveis que são beneficiárias do programa. Nos modelos de impacto (n=3446), as exposições principais foram o recebimento do benefício: beneficiário em 2010, no período de 2004-2010; o valor médio mensal recebido e o tempo de recebimento. Foram gerados modelos de regressão linear para os desfechos score-Z do índice de massa corporal por idade (IMC/I), percentual e índice de massa gorda (IMG), e percentual e índice de massa livre de gordura (IMLG); e de Poisson, com ajuste robusto, para o desfecho obesidade (score-Z IMC/I 2), todos estratificados por sexo. As informações antropométricas e de composição corporal (BOD POD) foram obtidas do acompanhamento aos 6-7 anos de idade. Potenciais fatores de confusão foram identificados por modelo hierárquico e por um diagrama causal (DAG). Para analisar os impactos foram usadas como medidas de efeito a diferença de médias na regressão linear múltipla (IMC/I, por cento MG, IMG, por cento MLG e IMLG, variáveis contínuas) e a razão de prevalência (obesidade, variável binária). Para permanecer no modelo, considerou-se valor p0,20. A análise dos dados foi realizada por meio do software STATA. Resultados: Entre 2004-2010, a proporção de famílias beneficiárias na coorte aumentou (11 por cento para 34 por cento ) enquanto, de acordo com a renda familiar, a proporção de famílias elegíveis diminui (29 por cento para 16 por cento ). No mesmo período, a cobertura do programa aumentou tanto pela renda familiar quanto pelo IEN. Já a focalização caiu de 78 por cento para 32 por cento de acordo com a renda familiar e, de acordo com o IEN, manteve-se em 37 por cento . A média (não ajustada) de IMC e de MG dos não beneficiários foi superior a dos não beneficiários tanto em meninos quanto em meninas. Meninos do 3º tercil de valor per capita recebido e meninas com menos de 7 meses de benefício em 2010 tiveram IMC maior do que, respectivamente, aqueles dos demais tercis e daquelas com mais de 7 meses de benefício em 2010; esse padrão foi semelhante para obesidade. Meninas não beneficiárias tiveram MG maior do que as beneficiárias e superior também aos meninos, independente de ser beneficiário ou não. Em relação à MLG observou-se um comportamento contrário, no qual meninas beneficiárias tiveram maior MLG, quando comparadas com meninas não beneficiárias e, meninos quando comparados com meninas. Nos modelos de regressão ajustados, não houve diferença significativa entre beneficiários e não beneficiários em nenhum desfecho. Conclusões: De acordo com os resultados, as famílias que receberam maiores valores per capita parecem incluir crianças com maior média de IMC. O programa, nessa análise, parece não ter impacto sobre a composição corporal das crianças, nem em termos de massa gorda, tampouco em termos de massa livre de gordura.

Hα emission fluxes and lithium abundances of low-mass stars in the young open cluster IC 4665

As part of a long term effort to understand pre-main sequence Li burning, we have obtained high resolution spectroscopic observations of 14 late type stars (G0-M1) in the young open cluster IC 4665. Most of the stars have Hα filled-in and Li I absorption, as expected for their young age. From the equivalent widths of Hα emission excess (obtained using the spectral subtraction technique) and the the Li i λ6708 feature, we have derived Hα emission fluxes and photospheric Li abundances. The mean Li abundance of IC 4665 solar-type stars is log N(Li) = 3.1; the same as in other young clusters (α Per, Pleiades) and T Tauri stars. Our results support the conclusions from previous works that PMS Li depletion is very small for masses ∼ 1 M_⨀ . Among the IC 4665 late-G and early K-type stars, there is a spread in Li abundances of about one order of magnitude. The Li-poor IC 4665 members have low Hα excess and vsini≤10. Hence, the Li-activity-rotation connection which has been clearly established in the Pleiades also seems to hold in IC 4665. One M-type IC 4665 star that we have observed does not show Li, implying a very efficient Li depletion as observed in α Per stars of the same spectral type. The level of chromospheric activity and Li depletion among the low-mass stars of IC 4665 is similar to that in the Pleiades. In fact, we note that the Li abundance distributions in several young clusters (α Per, Pleiades, IC 2391, IC 4665) and in post T Tauri stars are strikingly similar. This result suggests that Hα emission and Li abundance not well correlated with age for low-mass stars between 20 and 100 Myr old. We argue that a finer age indicator, the ''LL-clock'', would be the luminosity at which the transition between efficient Li depletion and preservation takes place for fully convective objects. The LL-clock could allow in the near future to derive the relative ages of young open clusters, and clarify the study of PMS evolution of cool stars.

Accreting magnetars: a new type of high-mass X-ray binaries?

The discovery of very slow pulsations (Pspin =5560 s) has solved the long-standing question of the nature of the compact object in the high-mass X-ray binary 4U 2206+54 but has posed new ones. According to spin evolutionary models in close binary systems, such slow pulsations require a neutron star magnetic field strength larger than the quantum critical value of 4.4 × 1013 G, suggesting the presence of a magnetar. We present the first XMM–Newton observations of 4U 2206+54 and investigate its spin evolution. We find that the observed spin-down rate agrees with the magnetar scenario. We analyse Integral Spacecraft Gamma-Ray Imager (ISGRI)/INTErnational Gamma-RAy Laboratory (INTEGRAL) observations of 4U 2206+54 to search for the previously suggested cyclotron resonance scattering feature at ∼30 keV. We do not find a clear indication of the presence of the line, although certain spectra display shallow dips, not always at 30 keV. The association of these dips with a cyclotron line is very dubious because of its apparent transient nature. We also investigate the energy spectrum of 4U 2206+54 in the energy range 0.3–10 keV with unprecedented detail and report for the first time the detection of very weak 6.5 keV fluorescence iron lines. The photoelectric absorption is consistent with the interstellar value, indicating very small amount of local matter, which would explain the weakness of the florescence lines. The lack of matter locally to the source may be the consequence of the relatively large orbital separation of the two components of the binary. The wind would be too tenuous in the vicinity of the neutron star.

Determination of fat-soluble vitamins in vegetable oils through microwave-assisted high-performance liquid chromatography

In this manuscript, a study of the effect of microwave radiation on the high-performance liquid chromatography separation of tocopherols and vitamin K1 was conducted. The novelty of the application was the use of a relatively low polarity mobile phase in which the dielectric heating effect was minimized to evaluate the nonthermal effect of the microwave radiation over the separation process. Results obtained show that microwave-assisted high-performance liquid chromatography had a shorter analysis time from 31.5 to 13.3 min when the lowest microwave power was used. Moreover, narrower peaks were obtained; hence the separation was more efficient maintaining or even increasing the resolution between the peaks. This result confirms that the increase in mobile phase temperature is not the only variable for improving the separation process but also other nonthermal processes must intervene. Fluorescence detection demonstrated better signal-to-noise compared to photodiode arrayed detection mainly due to the independent effect of microwave pulses on the baseline noise, but photodiode array detection was finally chosen as it allowed a simultaneous detection of nonfluorescent compounds. Finally, a determination of the content of the vitamin E homologs was carried out in different vegetable oils. Results were coherent with those found in the literature.