131 research outputs found

    Brain development in the yellow fever mosquito Aedes aegypti: a comparative immunocytochemical analysis using cross-reacting antibodies from Drosophila melanogaster

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    Considerable effort has been directed towards understanding the organization and function of peripheral and central nervous system of disease vector mosquitoes such as Aedes aegypti. To date, all of these investigations have been carried out on adults but none of the studies addressed the development of the nervous system during the larval and pupal stages in mosquitoes. Here, we first screen a set of 30 antibodies, which have been used to study brain development in Drosophila, and identify 13 of them cross-reacting and labeling epitopes in the developing brain of Aedes. We then use the identified antibodies in immunolabeling studies to characterize general neuroanatomical features of the developing brain and compare them with the well-studied model system, Drosophila melanogaster, in larval, pupal, and adult stages. Furthermore, we use immunolabeling to document the development of specific components of the Aedes brain, namely the optic lobes, the subesophageal neuropil, and serotonergic system of the subesophageal neuropil in more detail. Our study reveals prominent differences in the developing brain in the larval stage as compared to the pupal (and adult) stage of Aedes. The results also uncover interesting similarities and marked differences in brain development of Aedes as compared to Drosophila. Taken together, this investigation forms the basis for future cellular and molecular investigations of brain development in this important disease vecto

    Lagerung von aus Buffy Coats gewonnenen Leukozytenkonzentraten über 72 Stunden

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    Ziel der Arbeit sollte sein, verschiedene Lagerungsbedingungen zu vergleichen und eine Methode zu finden, Granulozyten, die aus Buffy Coats gewonnen wurden, bis zu 72 Stunden bei bestmöglichem Erhalt ihrer Vitalität und Funktionalität lagern zu können. Dazu wurden 23 Buffy Coats aufgereinigt und 20 verschiedene Lagerungsbedingungen verglichen. Es wurden direkt nach Aufreinigung und alle 24 Stunden über 72 Stunden Test bezüglich Vitalität und Funktionaliät der neutrophilen Granulozyten durchgeführt (Diff-BB, Trypanblau-Test, Oxyburst durch Chemilumineszenz, Durchflusszytometrie, BGA)

    Prevalence and Infection Intensity of Human and Animal Tungiasis in Napak District, Karamoja, Northeastern Uganda

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    Tungiasis is an important but highly neglected cause of morbidity in resource-poor communities in Latin America and sub-Saharan Africa. Data upon which implementation of control measures can be based are scarce. Before piloting an integrated tungiasis control program in three parishes of Napak district, Uganda, a cross-sectional survey involving the systematic examination of humans and domestic mammals was implemented to establish the occurrence patterns of tungiasis. The study population was 5482 residents, of which 4035 (73.6%) participated in the study. The prevalence of tungiasis in humans was 62.8% (95% CI: 61.3–64.3%), with slightly more males than females affected (p = 0.01). Age-specific prevalence and intensity of human tungiasis followed an S-curve pattern, with children of 5–14 years and the elderly (≥60 years) being the most affected. Half of all lesions (50%) had been manipulated by sharp objects. The prevalence of tungiasis in animals was lower (14.2%, 95% CI: 10.9–18.0) than that of humans (p < 0.001). Animal tungiasis occurred in decreasing order of frequency in pigs (80%), dogs (24%), goats (16.3%), cats (8.1%) and sheep (4.9%). In conclusion, human tungiasis was highly prevalent but animal infections were comparatively few in the study area. Nevertheless, effective control measures should be based on One Health principles

    Дослідження впливу фосфатів на біопродуктивність Chlorella vulgaris

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    Підвищений вміст фосфору у водоймах є основною причиною їхньої евтрофікації. Евтрофікація – процес зростання водної рослинності, який відбувається внаслідок перевищення балансу поживних речовин. Він супроводжується надмірним розвитком водоростей, особливо зелених, синьо-зелених і діатомових, переважанням небажаних видів планктону, порушенням життєдіяльності риб. Продукти метаболізму водоростей дають воді неприємний запах, можуть викликати шкірні алергічні реакції і шлунково-кишкові захворювання у людей і тварин. Після відмирання водорості виділяють у воду поліпептиди, аміак і проміжні продукти білкового розпаду. Це може призводити до підвищення вмісту фенолів, які мають канцерогенні властивості

    Fibroblast Growth Factor 21 Response in a Preclinical Alcohol Model of Acute-on-Chronic Liver Injury

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    Background and Aims: Fibroblast growth factor (FGF) 21 has recently been shown to play a potential role in bile acid metabolism. We aimed to investigate the FGF21 response in an ethanol-induced acute-on-chronic liver injury (ACLI) model in Abcb4−/− mice with deficiency of the hepatobiliary phospholipid transporter. Methods: Total RNA was extracted from wild-type (WT, C57BL/6J) and Abcb4−/ − (KO) mice, which were either fed a control diet (WT-Cont and KO-Cont groups; n = 28/group) or ethanol diet, followed by an acute ethanol binge (WT-EtOH and KOEtOH groups; n = 28/group). A total of 58 human subjects were recruited into the study, including patients with alcohol-associated liver disease (AALD; n = 31) and healthy controls (n = 27). The hepatic and ileal expressions of genes involved in bile acid metabolism, plasma FGF levels, and bile acid and its precursors 7α- and 27-hydroxycholesterol (7α- and 27-OHC) concentrations were determined. Primary mouse hepatocytes were isolated for cell culture experiments. Results: Alcohol feeding significantly induced plasma FGF21 and decreased hepatic Cyp7a1 levels. Hepatic expression levels of Fibroblast growth factor receptor 1 (Fgfr1), Fgfr4, Farnesoid X-activated receptor (Fxr), and Small heterodimer partner (Shp) and plasma FGF15/FGF19 levels did not differ with alcohol challenge. Exogenous FGF21 treatment suppressed Cyp7a1 in a dose-dependent manner in vitro. AALD patients showed markedly higher FGF21 and lower 7α-OHC plasma levels while FGF19 did not differ. Conclusions: The simultaneous upregulation of FGF21 and downregulation of Cyp7a1 expressions upon chronic plus binge alcohol feeding together with the invariant plasma FGF15 and hepatic Shp and Fxr levels suggest the presence of a direct regulatory mechanism of FGF21 on bile acid homeostasis through inhibition of CYP7A1 by an FGF15-independent pathway in this ACLI model. Lay Summary: Alcohol challenge results in the upregulation of FGF21 and repression of Cyp7a1 expressions while circulating FGF15 and hepatic Shp and Fxr levels remain constant both in healthy and pre-injured livers, suggesting the presence of an alternative FGF15-independent regulatory mechanism of FGF21 on bile acid homeostasis through the inhibition of Cyp7a1

    Identification of a Pseudomonas aeruginosa PAO1 DNA methyltransferase, its Targets, and physiological roles

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    DNA methylation is widespread among prokaryotes, and most DNA methylation reactions are catalyzed by adenine DNA methyltransferases, which are part of restriction-modification (R-M) systems. R-M systems are known for their role in the defense against foreign DNA; however, DNA methyltransferases also play functional roles in gene regulation. In this study, we used single-molecule real-time (SMRT) sequencing to uncover the genome-wide DNA methylation pattern in the opportunistic pathogen Pseudomonas aeruginosa PAO1. We identified a conserved sequence motif targeted by an adenine methyltransferase of a type I R-M system and quantified the presence of N(6)-methyladenine using liquid chromatography-tandem mass spectrometry (LC-MS/MS). Changes in the PAO1 methylation status were dependent on growth conditions and affected P. aeruginosa pathogenicity in a Galleria mellonella infection model. Furthermore, we found that methylated motifs in promoter regions led to shifts in sense and antisense gene expression, emphasizing the role of enzymatic DNA methylation as an epigenetic control of phenotypic traits in P. aeruginosa Since the DNA methylation enzymes are not encoded in the core genome, our findings illustrate how the acquisition of accessory genes can shape the global P. aeruginosa transcriptome and thus may facilitate adaptation to new and challenging habitats.IMPORTANCE With the introduction of advanced technologies, epigenetic regulation by DNA methyltransferases in bacteria has become a subject of intense studies. Here we identified an adenosine DNA methyltransferase in the opportunistic pathogen Pseudomonas aeruginosa PAO1, which is responsible for DNA methylation of a conserved sequence motif. The methylation level of all target sequences throughout the PAO1 genome was approximated to be in the range of 65 to 85% and was dependent on growth conditions. Inactivation of the methyltransferase revealed an attenuated-virulence phenotype in the Galleria mellonella infection model. Furthermore, differential expression of more than 90 genes was detected, including the small regulatory RNA prrF1, which contributes to a global iron-sparing response via the repression of a set of gene targets. Our finding of a methylation-dependent repression of the antisense transcript of the prrF1 small regulatory RNA significantly expands our understanding of the regulatory mechanisms underlying active DNA methylation in bacteria

    SAS-1 Is a C2 Domain Protein Critical for Centriole Integrity in C. elegans

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    Centrioles are microtubule-based organelles important for the formation of cilia, flagella and centrosomes. Despite progress in understanding the underlying assembly mechanisms, how centriole integrity is ensured is incompletely understood, including in sperm cells, where such integrity is particularly critical. We identified C. elegans sas-1 in a genetic screen as a locus required for bipolar spindle assembly in the early embryo. Our analysis reveals that sperm-derived sas-1 mutant centrioles lose their integrity shortly after fertilization, and that a related defect occurs when maternal sas-1 function is lacking. We establish that sas-1 encodes a C2 domain containing protein that localizes to centrioles in C. elegans, and which can bind and stabilize microtubules when expressed in human cells. Moreover, we uncover that SAS-1 is related to C2CD3, a protein required for complete centriole formation in human cells and affected in a type of oral-facial-digital (OFD) syndrome
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