Assessment of Blood Contamination in Biological Fluids Using MALDI-TOF MS

Biological fluid sample collection often includes the risk of blood contamination that may alter the proteomic profile of biological fluid. In proteomics studies, exclusion of contaminated samples is usually based on visual inspection and counting of red blood cells in the sample; analysis of specific blood derived proteins is less used. To fill the gap, we developed a fast and sensitive method for ascertainment of blood contamination in crude biological fluids, based on specific blood-derived protein, hemoglobin detection by MALDI-TOF MS. The MALDI-TOF MS based method allows detection of trace hemoglobin with the detection limit of 0.12 nM. UV-spectrometry, which was used as reference method, was found to be less sensitive. The main advantages of the presented method are that it is fast, effective, sensitive, requires very small sample amount and can be applied for detection of blood contamination in various biological fluids collected for proteomics studies. Method applicability was tested on human cerebrospinal and follicular fluid, which proteomes generally do not contain hemoglobin, however, which possess high risk for blood contamination. Present method successfully detected the blood contamination in 12 % of cerebrospinal fluid and 24 % of follicular fluid samples. High percentage of contaminated samples accentuates the need for initial inspection of proteomic samples to avoid incorrect results from blood proteome overlap.     

New insights into halophilic prokaryotes isolated from salting–ripening anchovies (Engraulis anchoita) process focused on histamine-degrading strains

Salted and ripened fish foods are susceptible to cause histamine poisoning. The present study focuses on microbial histamine degradation from high salted fermented fishery products to deepen our understanding about this new and growing field of research. As a result of this first study related to salted–ripened anchovies (Engraulis anchoita), fifty seven moderate and extreme halophilic microbial isolates from salt and salted–ripened anchovy processes were characterized in terms of their phenotype and histamine-degrading capacity. Only 7%—4 isolates—were able to degrade histamine. None of the histamine-degrading isolates presented proteolytic and/or lipolytic activity. One of them designated A18 was chemotactic toward histamine, an interesting property not previously reported for that chemoattractant. However, the S18 and A18 isolates, genotypically identified as Halobacterium sp. and Halomonas sp. respectively, produced indole and/or H₂S, both undesirable characteristics associated to off-flavors occurrence. On the other hand, A28 and S20, identified as Halovibrio sp. and Halobacterium sp. respectively, presented desirable properties, such as cytochrome oxidase and catalase activity, and non-production of H₂S and indole. These strains also showed characteristics previously reported as dominant in the ripened stage. The results are promising, and A28 and S20 may have the desirable features to improve the anchovy salting–ripening process.

     

Ethinylestradiol and its effects on the macrophages in the prostate of adult and senile gerbils

Prenatal and neonatal exposure to estrogenic compounds, such as ethinylestradiol (EE), promotes a variety of developmental disorders, including malformations and alterations in the morphology of glands, such as the prostate gland. Therefore, the aim of this study was to evaluate the morphological effects of neonatal exposure to EE on prostatic tissue and on the identification and quantification of gerbil gland macrophages in adult and senile Mongolian gerbils. The animals were exposed to EE (10 μg/kg/day) and to the vehicle, mineral oil (100 μL) (control group) during the first 10 days of postnatal life (lactation period). Adult gerbils were euthanized at 120 days and senile gerbils at 12 months of age. Our findings permitted verification of the presence of areas with proliferative foci in the prostate glandular portions in the adult and senile animals exposed to EE. There was also an increase in macrophages in the prostate tissue of adult and senile gerbils; these cell types alter the stromal microenvironment and possibly modify the interactions between the epithelium and stroma. Neonatal exposure to EE changes the pattern of prostatic development, leading to alterations in the arrangement of cells, including macrophages, and may be related to the onset of proliferative disorders in the prostate of adult gerbils and during aging.     

Liana abundance and diversity increase with rainfall seasonality along a precipitation gradient in Panama

In tropical regions, rainfall gradients often explain the abundance and distribution of plant species. For example, many tree and liana species adapted to seasonal drought are more abundant and diverse in seasonally-dry forests, characterized by long periods of seasonal water deficit. Mean annual precipitation (MAP) is commonly used to explain plant distributions across climate gradients. However, the relationship between MAP and plant distribution is often weak, raising the question of whether other seasonal precipitation patterns better explain plant distributions in seasonally-dry forests. In this study, we examine the relationship between liana abundance and multiple metrics of seasonal and annual rainfall distribution to test the hypothesis that liana density and diversity increase with increasing seasonal drought along a rainfall gradient across the isthmus of Panama. We found that a normalized seasonality index, which combines MAP and the variability of monthly rainfall throughout the year, was a significant predictor of both liana density and species richness, whereas MAP, rainfall seasonality and the mean dry season precipitation (MDP) were far weaker predictors. The strong response of lianas to the normalized seasonality index indicates that, in addition to the total annual amount of rainfall, how rainfall is distributed throughout the year is an important determinant of the hydrological conditions that favor liana proliferation. Our findings imply that changes in annual rainfall and rainfall seasonality will determine the future distribution and abundance of lianas. Models that aim to predict future plant diversity, distribution, and abundance may need to move beyond MAP to a more detailed understanding of rainfall variability at sub-annual timescales.     

Historical demography and climate driven distributional changes in a widespread Neotropical freshwater species with high economic importance

The Neotropical region exhibits the greatest worldwide diversity and the diversification history of several clades is related to the puzzling geomorphologic and climatic history of this region. The freshwater Amazon ecoregion contains the main hydrographic basins of the Neotropical region that are highly dendritic and ecologically diverse. It contains a rich and endemic fish fauna, including one of its most iconic and economically important representatives, the bony-tongue Arapaima gigas (Teleostei, Osteoglossiformes). Here, we evaluated the projected distribution of the genus in different historical periods (Present, Last Glacial Maximum, Last Interglacial Maximum and Near Future) and interpreted these results in light of the genomic diversity and modeled historical demography. For that, we combined species distribution models, population genetic analysis using SNPs and deep learning model selection. We analyzed a representative sample of the genus from the two basins where it naturally occurs, four localities in the Amazon (Am) and three in the Tocantins-Araguaia (To-Ar) basin, as well as individuals from three fish farms. We inferred a potentially smaller distribution in the glacial period, with a possible refuge in central Am. Our genetic data agrees with this result, suggesting a higher level of genetic diversity in the Am basin, compared to that observed in To-Ar. Our deep learning model comparison indicated that the To-Ar basin was colonized by the population from the Am basin. Considering a global warming scenario in the near future, A. gigas could reach an even larger range, especially if anthropogenic related dispersal occurs, potentially invading new areas and impacting their communities.