Identifying metalloproteins through X-ray fluorescence mapping and mass spectrometry

Metals are critical and dynamic components of biochemistry. To understand their roles, we greatly need tools to identify the ligands that bind them within the complexity of natural systems. This work describes the development of methods that not only detect and distinguish metals, but also characterize the proteins that bind them. We describe an approach to expose, identify and quantify metalloproteins in complex mixtures by sequential non-denaturing 2D-gel electrophoresis (2D GE)/X-ray Fluorescence (XRF) and tandem mass spectrometry (MS/MS) in the same spot of a sample. We first apply the development of 2D GE/XRF to Shewanella oneidensis MR-1, a well-studied system, and verify our electrophoretic approach. Then, we identified a novel periplasmic zinc protein in Pseudomonas aeruginosa PAO1 through 2D GE/XRF followed by MS/MS. The identity and function of this protein was verified through a gene mutation experiment.Fil: Raimunda, Daniel Cesar. Worcester Polytechnic Institute; Estados UnidosFil: Khare, Tripti. Argonne National Laboratory; Estados UnidosFil: Giometti, Carol. Argonne National Laboratory; Estados UnidosFil: Vogt, Stefan. Argonne National Laboratory; Estados UnidosFil: Argüello, José M.. Worcester Polytechnic Institute; Estados UnidosFil: Finney, Lydia. Argonne National Laboratory; Estados Unido

PERFIL PROTEICO DO LÍQUIDO FOLICULAR COLETADO DE OVÁRIOS EM DIFERENTES FASES DO CICLO ESTRAL DE BOVINOS.

O microambiente do gameta feminino é o líquido folicular e, portanto as proteínas presentes no líquido folicular devem interferir na qualidade do oócito. Com o objetivo de verificar o perfil proteico do líquido folicular de folículos de diferentes fases do ciclo estral de bovinos este experimento foi delineado. Para tanto, os líquidos foliculares provenientes de ovários de vacas de abatedouro foram alocados em 5 diferentes fases do ciclo estral, levando em conta a presença ou ausência do corpo lúteo (CL) e sua caracterização morfológica. “Pools” de líquido folicular foram coletados de folículos de 2 a 7 mm de ovários em 5 diferentes fases do ciclo estral (1=CL inicial, hemorrágico; 2=CL em desenvolvimento; 3=CL maduro; 4=CL em regressão e 5=ausência de CL). O perfil proteico foi avaliado por eletroforese em gel de poliacrilamida e determinado em porcentagens nas amostras de cada fase. Os polipeptídeos de 35, 32, 30 e 18 KDa variam consideravelmente nas amostras de diferentes fases do ciclo estral. O polipeptídeo de mesmo peso molecular que o IGFBP-2 está mais presente nas amostras de líquido folicular ao final do ciclo estral. Maior número de polipeptídeos é observado em folículos antrais pequenos de ovários em que o CL está em regressão

Herpetic Meningoencephalitis by Vertical Transmission: A Case Report and Systematic Review

Objective: To report a case of vertical herpetic meningoencephalitis. Results: The involvement of the central nervous system (CNS) in infection by HSV (herpes simplex virus), HSV-1 or HSV-2, causes an acute inflammatory process in the brain parenchyma, leading to herpetic encephalitis. It is a feared form of the disease due to its severity and its high rate of morbidity and mortality. Its rapid fatal progression can be prevented from early suspicion and treatment, which is essential when taking into account their neurological sequelae since survivors have motor sequelae, behavioral disorders, or epilepsy. The present work reports the case of a newborn male with spontaneous vaginal delivery who, at 19 days of age, started to experience fever, irritability, difficulty in eating, spasms, tremors of the upper limbs, deviation of the eyes, and seizures of difficult to control, together with CFE and serological changes, in addition to imaging tests compatible with herpetic meningoencephalitis, progressing with a very serious evolution despite the institution of specific treatment for CNS herpetic infection, evolving with important neurological sequelae. Conclusion: The sequels resulting from herpetic encephalitis not properly diagnosed, or even late, leads from severe neurological damage to death. Therefore, it is extremely important to start empirical treatment with antiviral drugs to reduce the sequelae mentioned above

Sono utili i Translation Studies per la pratica della traduzione?

The starting point for the present contribution is that professional translators as well as editors tend to consider the theoretical debate on translation with mistrust. By looking at the translation process in its various phases, this paper aims to demonstrate how theory does, as a matter of fact, provide a 'meta-language' that can be of great help in the training of prospective translators, while at the same time offering useful tools for an evaluation of the translated text that goes beyond a merely impressionistic critique, based on individual taste. Furthermore, this paper discusses a number of theoretical concepts developed in the field of translation theory that can be of use to the translator's analytical work before, during, and after his/her activity of transcodification

Resposta imune humoral em bovinos da raça nelore vacinados contra a raiva e suplementados com crômio orgânico

With the objective of to evaluate the effect of the addition of orgainc chromium in the humoral immune response against rabies, five Neloreþs bovine breed groups with 15 animals each, were supplementeds with chromium yeast being the group GC (control), GV (vaccinated and without chromium), GV+8,5 (vaccinated + 8,5 ppm of chromium), GV+17 (vaccinated + 17 ppm of chromium) and GV+34 (vaccinated +34 ppm of chromium). Serum samples of this animals were analyseds with serum neutralization test (SN) in albines mices in the days 30,60 and 90 pos -vaccination. Simultaneously, serum concentrations of the chromium were determinated for analysis with spectrophotometer of atomic absorption in graphite oven and analysis of the cortisol was achieved through of the radioimmunoassay.These results allows to conclude that the animals of the group GV+34 have persistence of the protector's antibody titulation anti rabies until 90 days pos- vaccination. This response does not dependt of the cortisol levels, since the chromium donþt show to be effective to decrease its levels

Suplementação com levedura de Crômio eleva a concentração sérica de Crômio em bovinos

This study evaluated the effect of daily chromium supplementation at growing doses on serum Cr levels in cattle. Sixty Nellore heifers 2 to 5 years old were randomly assigned to one of four groups (15 heifers/group) fed non-fertilized Brachiaria decumbens pasture. In the control group (Gc), the heifers received a mineral mixture without Cr. In the other groups, heifers received 0.59, 1.19 or 2.38 mg Cr+3/animal/day, groups G8,5, G17 and G34, respectively. Blood samples were collected on days 0, 30 and 60. Serum concentration of chromium was determined by graphite furnace atomic absorption spectrophotometer. The chromium source used was the chromium yeast. The results do not support any significant difference (p>0.05) among serum chromium levels of heifers in groups Gc, G8,5, G17 and G34 at a same observation day. Only effect of supplementation time was detected in group G34. In this group there was significant increase (p0,05) na concentração sérica de crômio das novilhas entre os grupos G8,5, G17 e G34, num mesmo dia de observação. O único efeito observado foi do tempo de suplementação com crômio no o grupo G34. Neste grupo a concentração sérica de crômio apresentou aumento significativo (p<0,05) 60 dias após o início da suplementação. Conclui-se a levedura de Cr pode ser utilizada como fonte deste elemento mineral para os bovinos e a suplementação com 2,38 mg/bovino/dia foi a dose que mais elevou a concentração sérica de Cr nos bovinos da raça Nelore durante o período experimental

Structural changes and crosslinking modulated functional properties of oxi-HA/ADH hydrogels useful for regenerative purposes

Partially oxidized hyaluronic acid (oxi-HA) crosslinked with adipic acid dihydrazide (ADH) has been explored as a promising injectable biomaterial for cell-based therapy and regenerative purposes. However, there is a lack of basic scientific studies and medical applications to support the technological approaches and optimizations of these biomaterials. Thus, we provided a relationship connecting the structural changes to the functional properties, which are modulated by the oxi-HA oxidation degree and ADH crosslinking. Oxi-HA was obtained from the reaction with sodium periodate (NaIO4), which oxidizes the proximal hydroxyl in aldehydes groups by opening the sugar ring and promoting macromolecular compaction. The results showed that the oxi-HA oxidation degree caused the structural changes that increased the zeta potential and decreased the hydrodynamic volume and viscosity. Therefore, the extrusion force, injectability, and ADH crosslinking were benefited by the click chemistry. Additionally, the ADH concentration modulated the functional properties such as the swelling, stability, gelation time and thermosensitivity of the oxi-HA/ADH hydrogels. These results are relevant for supporting the design and optimization of novel hyaluronic acid-based formulations for applications in the broad field of tissue regeneration121CONSELHO NACIONAL DE DESENVOLVIMENTO CIENTÍFICO E TECNOLÓGICO - CNPQFUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULO - FAPESP140924/2017-5; 153611/2016-22016/10132-