Ultra-estrutura de glândulas abdominais tegumentares em Oxaea flavescens (Hymenoptera, Andrenidae, Oxaeinae)

The sternal glands of the abdomen of Oxaea flavescens (Klug, 1807) consist of class III glandular cells around a reservoir constituted by branched folds of the intersegmental membrane of segments III, IV and V. The gland cells are rich in rough endoplasmic reticulum and produce a secretion with mucous aspect. The treatment with oxidated osmium and ruthenium red showed numerous Golgi regions in the cell and carbohydrates absorption from the haemolymph, respectively. The high degree of development of the glands suggests an important function to the species, although still unknown.

Histologia e ultra-estrutura do vaso dorsal de Scaptotrigona postica (Hymenoptera, Apidae) em operárias e rainhas de diferentes idades

O vaso dorsal é um órgão tubular localizado na região mediano-dorsal do corpo dos insetos, abaixo do tegumento. Fez-se um estudo de microscopia de luz e eletrônica de transmissão da porção abdominal do vaso dorsal, o coração, em uma espécie de abelha indígena. Foram estudadas operárias e rainhas em diferentes idades. O coração está localizado no sinus pericárdico. A parede cardíaca é formada por fibras musculares estriadas e apresenta aberturas ou ostíolos providos de válvulas. A fibra cardíaca contém miofibrilas arranjadas irregularmente, núcleos alongados ou redondos, mitocôndrias grandes e numerosas, e depósitos de glicogênio. Em operárias e rainhas longevas, as fibras encontram-se em degeneração, evidenciada por vacúolos autofágicos, alterações mitocondriais e acúmulo de corpos mielínicos. Em conclusão, o coração de Scaptotrigona postica é semelhante ao de outros insetos estudados. As alterações encontradas estão relacionadas ao processo de envelhecimento e mantêm relação temporal com a expectativa de vida da casta.

Espermiogênese em Eupemphix nattereri (Anura, Leiuperidae): aspectos ultra-estruturais

A maturação dos espermatozóides envolve um extenso e complexo processo que começa com a proliferação e diferenciação das espermatogônias, passa pela meiose e finaliza com a espermiogênese. Nessa fase, eventos envolvendo alterações morfológicas e bioquímicas transformam espermátides em espermatozóides. Aspectos ultra-estruturais da espermiogênese e do espermatozóide do anuro Eupemphix nattereri (Steindachner, 1863) foram analisados através de microscopia eletrônica de transmissão. A espermiogênese envolve condensação da cromatina e alongamento nuclear, com visível eliminação de citoplasma. Nesse estágio, grande quantidade de microtúbulos e glicogênio podem ser visualizados no citoplasma das células de Sertoli, rodeando cada espermátide. O espermatozóide é fusiforme e o acrossomo forma uma capa na região anterior do núcleo. A bainha mitocondrial é encontrada ao redor da porção proximal da cauda. A cauda apresenta o axonema com o modelo 9+2, uma fibra axonemal, a membrana ondulante e ausência de bastão axial. Esta organização apresenta algumas similaridades com espécies do gênero Physalaemus (Leiuperidae) como P. biligonigerus (Cope, 1861), P. gracilis (Boulenger, 1883) e P. fuscomaculatus (Steindachner, 1864).

Estudos citológicos e citoquímicos em Stenophoridae Crawley, 1903 (Eugregarinidae, Protozoa) II - ultra-estrutura

1 - Indivíduos de Stenophora juli (Frantzius, 1848) Labbé, 1899, parasitos de um Diplopoda, Rhinochricus padbergi Schubart. 1930 foram examinados em microscópia óptica e eletrônica. 2 - Os resultados do estudo citoquímico confirmam os dados obtidos por outros autores em outras espécies de gregarinas. 3 - Quanto à estrutura fina da morfologia celular foi examinada detalhadamente a película a qual apresenta cristas longitudinais de forma e estrutura complexas. 4 - No sulcos da película, entre as cristas, foram encontrados poros na membrana, por onde é realizada a secreção de muco. 5 - Aderente à película, pròpriamente dita foi encontrada, no deutomerito, uma camada homogênea de natureza desconhecida, abaixo da qual encontra-se o mionema. 6 - O septo que separa o proto do deutomerito é constituído por espêssa camada de mionemas incluindo numerosas mitocôndrias. 7 - O endoplasma é extremamente rico em granulações de paraglicogênio, aparecendo em menor quantidade os lipídeos. Observamos também mitocôndrias, retículo endoplasmático e o complexo de Golgi.

Segunda contribuição sobre as alterações ultra-estruturais dos hepatócitos na forma aguda (toxêmica) da esquistossomose mansoni

Estudamos as alterações ultra-estruturais dos hepatócitos na forma aguda, toxêmica, da esquistossomose, em cinco pacientes, membros de uma mesma família infectados em idênticas condições em um córrego existente próximo da lagoa de Pampulha, em Belo Horizonte (MG), e não tratados especificamente para a esquistossomose. Este estudo confirma os dados obtidos em trabalho anterior, em sete pacientes infectados no Município de Sabara (MG). Nos cinco casos, as alterações ultra-estruturais foram inespecíficas, pouco acentuadas, embora mais intensas do que as observadas anteriormente, e se caracterizaram sobretudo pelas modificações das organelas citoplasmáticas, explicando o freqüente encontro de células claras a microscopia óptica. A identificação de alguns granulomas a microscopia eletrônica permitiu verificar que estes mostram, no exstudato, granulócitos eosinófilos, macrófagos, plasmócitos, células epitelióides e mastócitos. Entre as células havia material amorfo e finos feixes de colágeno.

Ultra high performance supercritical fluid chromatography coupled with tandem mass spectrometry for screening of doping agents. II: Analysis of biological samples.

The potential and applicability of UHPSFC-MS/MS for anti-doping screening in urine samples were tested for the first time. For this purpose, a group of 110 doping agents with diverse physicochemical properties was analyzed using two separation techniques, namely UHPLC-MS/MS and UHPSFC-MS/MS in both ESI+ and ESI- modes. The two approaches were compared in terms of selectivity, sensitivity, linearity and matrix effects. As expected, very diverse retentions and selectivities were obtained in UHPLC and UHPSFC, proving a good complementarity of these analytical strategies. In both conditions, acceptable peak shapes and MS detection capabilities were obtained within 7min analysis time, enabling the application of these two methods for screening purposes. Method sensitivity was found comparable for 46% of tested compounds, while higher sensitivity was observed for 21% of tested compounds in UHPLC-MS/MS and for 32% in UHPSFC-MS/MS. The latter demonstrated a lower susceptibility to matrix effects, which were mostly observed as signal suppression. In the case of UHPLC-MS/MS, more serious matrix effects were observed, leading typically to signal enhancement and the matrix effect was also concentration dependent, i.e., more significant matrix effects occurred at the lowest concentrations.

Heterogeneity of atherosclerotic plaque phenotypes and composition in four different arterial beds: an intravascular ultrasound virtual histology study

Purpose: The purpose of this study was to compare the plaque morphology between coronary and peripheral arteries using intravascular ultrasound (IVUS). Methods: IVUS was performed in 68 patients with coronary and 93 with peripheral artery lesions (29 carotid, 50 renal, and 14 iliac). Plaques were classified as fibroatheroma (VH-FA) (further subclassified as thin-capped [VH-TCFA] and thick-capped [VH-ThCFA]), fibrocalcific plaque (VH-FC) and pathological intimal thickening (VH-PIT). Results: Plaque rupture (13% of coronary, 7% of carotid, 6% of renal, and 7% of iliac arteries; P=NS) and VH-TCFA (37% of coronary, 24% of carotid, 16% of renal, and 7% of iliac arteries; P=0.02) was observed in all arteries. Compared to coronary arteries, VH-FA was less frequently observed in renal (P<0.001) and iliac arteries (P<0.006), while VH-PIT and VH-FC were prevalent in both of these peripheral arteries. Lesions with positive remodeling demonstrated more characteristics of VH-FA in coronary, carotid, and renal arteries compared to those with intermediate/negative remodeling (all P<0.01). There was positive relationship between RI and percent necrotic core area in all four arteries. Conclusions: Atherosclerotic plaque phenotypes were heterogeneous among four different arteries. In contrast, the associations of remodeling mode with plaque phenotype and composition were similar among the various arterial beds.

Quantification of clenbuterol at trace level in human urine by ultra-high pressure liquid chromatography-tandem mass spectrometry.

Clenbuterol is a β2 agonist agent with anabolic properties given by the increase in the muscular mass in parallel to the decrease of the body fat. For this reason, the use of clenbuterol is forbidden by the World Anti-Doping Agency (WADA) in the practice of sport. This compound is of particular interest for anti-doping authorities and WADA-accredited laboratories due to the recent reporting of risk of unintentional doping following the eating of meat contaminated with traces of clenbuterol in some countries. In this work, the development and the validation of an ultra-high pressure liquid chromatography coupled to electrospray ionization tandem mass spectrometry (UHPLC-ESI-MS/MS) method for the quantification of clenbuterol in human urine is described. The analyte was extracted from urine samples by liquid-liquid extraction (LLE) in basic conditions using tert butyl-methyl ether (TBME) and analyzed by UHPLC-MS/MS with a linear gradient of acetonitrile in 9min only. The simple and rapid method presented here was validated in compliance with authority guidelines and showed a limit of quantification at 5pg/mL and a linearity range from 5pg/mL to 300pg/mL. Good trueness (85.8-105%), repeatability (5.7-10.6% RSD) and intermediate precision (5.9-14.9% RSD) results were obtained. The method was then applied to real samples from eighteen volunteers collecting urines after single oral doses administration (1, 5 and 10μg) of clenbuterol-enriched yogurts.

Ultra deep blue: the ultimate chess player

Projecte d'adaptació del programa GNU Chess al sistema de grid computing 'Condor'. I amb això, es planteja un estudi sobre els algorismes de cerca i la seva aplicació en entorns distribuïts. Una sèrie de proves sobre unes mostres de una partida d'escacs contra el propi GNU Chess ens ajuden a posar de relleu els avantatges i inconvenients de cada un dels algorismes proposats.

Multiplex ultra-performance liquid chromatography-tandem mass spectrometry method for simultaneous quantification in human plasma of fluconazole, itraconazole, hydroxyitraconazole, posaconazole, voriconazole, voriconazole-N-oxide, anidulafungin, and caspofungin.

Therapeutic drug monitoring (TDM) may contribute to optimizing the efficacy and safety of antifungal therapy because of the large variability in drug pharmacokinetics. Rapid, sensitive, and selective laboratory methods are needed for efficient TDM. Quantification of several antifungals in a single analytical run may best fulfill these requirements. We therefore developed a multiplex ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method requiring 100 μl of plasma for simultaneous quantification within 7 min of fluconazole, itraconazole, hydroxyitraconazole, posaconazole, voriconazole, voriconazole-N-oxide, caspofungin, and anidulafungin. Protein precipitation with acetonitrile was used in a single extraction procedure for eight analytes. After reverse-phase chromatographic separation, antifungals were quantified by electrospray ionization-triple-quadrupole mass spectrometry by selected reaction monitoring detection using the positive mode. Deuterated isotopic compounds of azole antifungals were used as internal standards. The method was validated based on FDA recommendations, including assessment of extraction yields, matrix effect variability (<9.2%), and analytical recovery (80.1 to 107%). The method is sensitive (lower limits of azole quantification, 0.01 to 0.1 μg/ml; those of echinocandin quantification, 0.06 to 0.1 μg/ml), accurate (intra- and interassay biases of -9.9 to +5% and -4.0 to +8.8%, respectively), and precise (intra- and interassay coefficients of variation of 1.2 to 11.1% and 1.2 to 8.9%, respectively) over clinical concentration ranges (upper limits of quantification, 5 to 50 μg/ml). Thus, we developed a simple, rapid, and robust multiplex UPLC-MS/MS assay for simultaneous quantification of plasma concentrations of six antifungals and two metabolites. This offers, by optimized and cost-effective lab resource utilization, an efficient tool for daily routine TDM aimed at maximizing the real-time efficacy and safety of different recommended single-drug antifungal regimens and combination salvage therapies, as well as a tool for clinical research.

Alterations of Neuromuscular Function after the World's Most Challenging Mountain Ultra-Marathon.

We investigated the physiological consequences of the most challenging mountain ultra-marathon (MUM) in the world: a 330-km trail run with 24000 m of positive and negative elevation change. Neuromuscular fatigue (NMF) was assessed before (Pre-), during (Mid-) and after (Post-) the MUM in experienced ultra-marathon runners (n = 15; finish time  = 122.43 hours ±17.21 hours) and in Pre- and Post- in a control group with a similar level of sleep deprivation (n = 8). Blood markers of muscle inflammation and damage were analyzed at Pre- and Post-. Mean ± SD maximal voluntary contraction force declined significantly at Mid- (-13±17% and -10±16%, P<0.05 for knee extensor, KE, and plantar flexor muscles, PF, respectively), and further decreased at Post- (-24±13% and -26±19%, P<0.01) with alteration of the central activation ratio (-24±24% and -28±34% between Pre- and Post-, P<0.05) in runners whereas these parameters did not change in the control group. Peripheral NMF markers such as 100 Hz doublet (KE: -18±18% and PF: -20±15%, P<0.01) and peak twitch (KE: -33±12%, P<0.001 and PF: -19±14%, P<0.01) were also altered in runners but not in controls. Post-MUM blood concentrations of creatine kinase (3719±3045 Ul·(1)), lactate dehydrogenase (1145±511 UI·L(-1)), C-Reactive Protein (13.1±7.5 mg·L(-1)) and myoglobin (449.3±338.2 µg·L(-1)) were higher (P<0.001) than at Pre- in runners but not in controls. Our findings revealed less neuromuscular fatigue, muscle damage and inflammation than in shorter MUMs. In conclusion, paradoxically, such extreme exercise seems to induce a relative muscle preservation process due likely to a protective anticipatory pacing strategy during the first half of MUM and sleep deprivation in the second half.