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There is no suitable vaccine against human visceral leishmaniasis (VL) and available drugs are toxic and/or present high cost. In this context, diagnostic tools should be improved for clinical management and epidemiological evaluation of disease. However, the variable sensitivity and/or specificity of the used antigens are limitations, showing the necessity to identify new molecules to be tested in a more sensitive and specific serology. In the present study, an immunoproteomics approach was performed in Leishmania infantum promastigotes and amastigotes employing sera samples from VL patients. Aiming to avoid undesired cross-reactivity in the serological assays, sera from Chagas disease patients and healthy subjects living in the endemic region of disease were also used in immunoblottings. The most reactive spots for VL samples were selected, and 29 and 21 proteins were identified in the promastigote and amastigote extracts, respectively. Two of them, endonuclease III and GTP-binding protein, were cloned, expressed, purified and tested in ELISA experiments against a large serological panel, and results showed high sensitivity and specificity values for the diagnosis of disease. In conclusion, the identified proteins could be considered in future studies as candidate antigens for the serodiagnosis of human VL.
Biogenic minerals are widely studied materials for their particular properties derived from their hierarchical structure, using building blocks with sizes spanning several orders of magnitude. These special features can be assessed with different analytical tools, and it is important to know their capabilities and limitations. In order to determine the hierarchical structure of the shells, the nacre and prismatic layers of two marine animals were characterized by infrared spectroscopy, X-ray diffraction, and scanning electron microscopy. Based on these assessments, we found that the combination of these three techniques is useful to describe each structure level, and to explain some of the unique properties observed in these natural materials.
Literature about fish kidney peroxisomes is scarce. To tackle this caveat, a stereological approach on renal peroxisome morphological parameters was performed for the first time in a fish, establishing correlations with maturation stages as it was previously done in brown trout liver. Three-year-old brown trout males and females were collected at the major seasons of their reproductive cycle. Trunk kidney was fixed and processed for catalase cytochemistry. Classical stereological methods were applied to electromicrographs to quantitate morphological parameters. Different seasonal variation patterns were observed between genders, and between renal proximal tubule segments I and II. In males, peroxisomes from proximal tubule segment II had a relatively higher volume and number in May, being individually bigger in February. Females presented similar trends, though with less marked variations. Overall, males and females did not show exactly the same seasonal patterns for most peroxisomal parameters, and no correlations were found between the latter and the gonado-somatic index (GSI). Hence, and despite the variations, the morphology of renal peroxisomes is not strictly correlated with gonad maturation kinetics, therefore suggesting that kidney peroxisome morphology is not seasonally modulated by sex steroids, like estradiol, as it seems to happen in liver peroxisomes.
The ultrastructural analysis of human oocytes at different maturation stages has only been descriptive. The aim of this study was to use a stereological approach to quantify the distribution of organelles in oocytes at prophase I (GV). Seven immature GV oocytes were processed for transmission electron microscopy and a classical manual stereological technique based on point-counting with an adequate stereological grid was used. The Kruskal–Wallis test and Mann–Whitney U-test with Bonferroni correction were used to compare the means of the relative volumes occupied by organelles in oocyte regions: cortex (C), subcortex (SC) and inner cytoplasm (IC). Here we first describe in GV oocytes very large vesicles of the smooth endoplasmic reticulum (SER), vesicles containing zona pellucida-like materials and coated vesicles. The most abundant organelles were the very large vesicles of the SER (6.9%), mitochondria (6.3%) and other SER vesicles (6.1%). Significant differences in organelle distribution were observed between ooplasm regions: cortical vesicles (C: 1.3% versus SC: 0.1%, IC: 0.1%, P = 0.001) and medium-sized vesicles containing zona pellucida-like materials (C: 0.2% versus SC: 0.02%, IC: 0%, P = 0.004) were mostly observed at the oocyte cortex, whereas mitochondria (C: 3.6% versus SC: 6.0%, IC: 7.2%, P = 0.005) were preferentially located in the subcortex and inner cytoplasm, and SER very large vesicles (IC: 10.1% versus C: 0.9%, SC: 1.67%, P = 0.001) in the oocyte inner cytoplasm. Further quantitative studies are needed in immature metaphase-I and mature metaphase-II oocytes, as well as analysis of correlations between ultrastructural and molecular data, to better understand human oocyte in vitro maturation.
An adult male specimen of Uca rapax (Crustacea, Decapoda, Ocypodidae), containing bilaterally hypertrophied chelipeds, was found in the mangrove area of the Rio Grande do Norte state, north-eastern Brazil. The chelipeds are subequal in size and similar to the major cheliped of normal male specimens of Uca rapax. This paper is the first record of the aforementioned anomaly for the species from the Brazilian coastline.
The porcellanid crab Porcellana sayana is, for the first time, reported to have a symbiotic association with the crab Stratiolibinia bellicosa. This is also the first record on the association between the porcelain crab P. sayana and a brachyuran crab on the Brazilian coastline. In addition, this is the second record of association of P. sayana with a Majoidea crab.
Shell bed levels in the Low Head Member of the early Oligocene Polonez Cove Formation at King George Island, West Antarctica, are re-interpreted based on sedimentological and taphonomic data. The highly fossiliferous Polonez Cove Formation is characterized by basal coastal marine sandstones, overlain by conglomerates and breccias deposited in fan-delta systems. The shell beds are mainly composed of pectinid bivalve shells of Leoclunipecten gazdzickii and occur in the basal portion of the Low Head Member. Three main episodes of bioclastic deposition are recorded. Although these shell beds were previously interpreted as shelly tempestites, we present an alternative explanation: the low fragmentation rates and low size sorting of the bioclasts resulted from winnowing due to tidal currents (background or diurnal condition) in the original bivalve habitat. The final deposition (episodic condition) was associated with subaqueous gravity driven flows. This new interpretation fits with the scenario of a prograding fan-delta front, which transported shell accumulations for short distances near the depositional site, possibly between fair-weather and storm wave bases. This work raises the notion that not every shell bed with similar sedimentological and taphonomic features (such as geometry, basal contact, degree of packing and shell orientation in the matrix) is made in the same way.
The role of activin-A in follicular development and on the mRNA expression levels of different genes in goat secondary follicles was evaluated. Goat secondary follicles (≥150 μm) were cultured for 18 days under control conditions or with the addition of either 50 or 100 ng/ml activin-A (Experiment 1). The mRNA levels for the genes that code for activin-A, ActR-IA, ActR-IB, ActR-IIA, ActR-IIB, follicle stimulating hormone receptor (FSH-R) and P450 aromatase were measured in each condition (Experiment 2). We observed that after 6 days of culture, the antrum formation rate was higher in cultures with added activin-A than in the cultured control (P < 0.05). The addition of 50 ng/ml activin-A increased the follicular growth rate in the final third of the culture (days 12–18), resulting in a higher percentage of meiosis resumption (P < 0.05). On day 6, the addition of activin-A (50 ng/ml) increased the levels of ActR-IA mRNA compared with the cultured control (P < 0.05). After 18 days, the addition of 50 ng/ml activin-A significantly increased the levels of its own mRNA compared with the non-cultured control. Moreover, this treatment reduced the mRNA levels of P450 aromatase in comparison with the cultured control (P < 0.05). Higher levels of P450 aromatase mRNA were found for both activin-A treatments compared with the non-cultured control (P < 0.05). No difference in estradiol levels was detected among any of the tested treatments. In conclusion, the addition of activin-A to culture medium stimulated early antrum formation as well as an increase in the daily follicular growth rate and the percentage of meiosis resumption.
Non gaussian sources of erros need to be taken into consideration when searching for planetary transits. Such phenomena are mostly caused by the impact of high energetic particles on the detector (Pinheiro da Silva et al. 2008). The detection efficiency of transits, therefor, depend significantly on the data quality and the algorithms utilized to deal with these errors sources. In this work we show that a modified detrend algorithm CDA (CoRoT Detrend Algorithm; Mislis et al. 2010) using a robust statistics and an empirical fit, instead of a polynomial one, can eliminate more efficiently gaps in the data and other long-term trends from the light-curve. Using this algorithm enables us to obtain a reconstructed light-curve with better signal-to-noise ratio that allows to improve the detection of exoplanet transits, although long term signals are destroyed. The results show that these modifications lead to an improved BLS (Box-fitting Least Squares; Kovács, Zucker & Mazeh 2002) algorithm spectrum. At the end we have compared our planetary search results with CoRoT (Convection, Rotation and planetary Transits) satellite chromatic light-curves available in the literature.
In this article, the sol-gel methodology was used for coating an acrylonitrile butadiene styrene (ABS) polymer prepared by the rapid prototyping technology with a colloid containing the europium III dipicolinic complex, which presents high emission when excited in the ultraviolet region. Either acid or base was used for treatment of the ABS polymer, with a view to activating its surface. The thermal analysis evidenced a residual mass after 600 °C, which indicated that the coating adhered to the substrate. X-ray diffraction analysis showed that the structure of the ABS polymer was not affected by the sol-gel treatment. The large band centered at 287 nm, ascribed to ligand-metal charge transfer, can be used to excite the europium III dipicolinic complex in the ultraviolet C and ultraviolet B regions. The emission appears in the characteristic red region of the electromagnetic spectrum. These results indicate that the obtained material is a candidate for use as ultraviolet sensor.