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Biology subjects

Thomas, L. S.

Publications and source records attributed to Thomas, L. S..

3 recordsLinked to original sources

A novel conserved protein in Streptococcus agalactiae, BvaP, is important for vaginal colonization and biofilm formation

Streptococcus agalactiae (Group B Strep, GBS) infections in neonates are often fatal and strongly associated with maternal GBS vaginal colonization. Here, we investigated the role of a previously uncharacterized protein, BvaP, in GBS vaginal colonization. BvaP was previously identified as the most highly upregulated gene in the GBS A909 transcriptome when comparing vaginal colonization to growth in liquid culture. We found that expression of BvaP affects GBS adherence to extracellular matrix components and human vaginal epithelial cells and a{Delta} bvaP mutant was significantly decreased in its ability to colonize the murine vaginal tract. Cellular morphological alterations such as changes in cell shape, chain length, and clumping were also observed in a knockout mutant strain. Given its high expression in vivo, high degree of conservation among GBS strains, and role in vaginal colonization, BvaP may be an eligible target for GBS vaccination and/or drug therapy. IMPORTANCENeonatal GBS disease is a major cause of morbidity and mortality and maternal vaginal colonization is the leading risk factor for disease. Colonization prevention would greatly impact rates of disease transmission, but vaccine development has stalled as capsular polysaccharide vaccines have low immunogenicity in vivo. While these vaccines are still in development, addition of a protein conjugate may prove fruitful in increasing immunogenicity and strain coverage across GBS serotypes. Previous research identified sak_1753 as a gene very highly upregulated gene during murine vaginal colonization. This study reveals that Sak_1753 is required to maintain proper GBS cellular morphology and colonization phenotypes and is required for in vivo vaginal colonization in a murine model. We have renamed Sak_1753 Group B Strep vaginal adherence protein (BvaP). The findings of this study indicate that BvaP is important in GBS colonization of the vaginal tract and may be a candidate for vaccine development.

microbiology↗

In vivo high-content screening in zebrafish for developmental nephrotoxicity of approved drugs

Despite widespread drug exposure, for example during gestation or in prematurely born children, organ-specific developmental toxicity of most drugs is poorly understood. Developmental and functional abnormalities are a major cause of kidney diseases during childhood; however, the potential causal relationship to exposure with nephrotoxic drugs during nephrogenesis is widely unknown. To identify developmental nephrotoxic drugs in a large scale, we established and performed an automated high-content screen to score for phenotypic renal alterations in the Tg(wt1b:EGFP) zebrafish line. During early nephrogenesis, embryos were exposed to a compound library of approved drugs. After treatment, embryos were aligned within microtiter plates using 3D-printed orientation tools enabling the robust acquisition of consistent dorsal views of pronephric kidneys by automated microscopy. To qualitatively and quantitatively score and visualize phenotypes, we developed software tools for the semi-automated analysis, processing and visualization of this large image-based dataset. Using this scoring scheme, we were able to categorize compounds based on their potential developmental nephrotoxic effects. About 10% of tested drugs induced pronephric phenotypes including glomerular and tubular malformations, or overall changes in kidney morphology. Major chemical compound groups identified to cause glomerular and tubular alterations included dihydropyridine derivatives, HMG CoA reductase inhibitors, fibrates, imidazole, benzimidazole and triazole derivatives, corticosteroids, glucocorticoids, acetic acid derivatives and propionic acid derivatives. In conclusion, the presented study demonstrates the large-scale screening of kidney-specific toxicity of approved drugs in a live vertebrate embryo. The associated technology and tool-sets can be easily adapted for other organ systems providing a unique platform for in vivo large-scale assessment of organ-specific developmental toxicity or other biomedical applications. Ultimately, the presented data and associated visualization and browsing tools provide a resource for potentially nephrotoxic drugs and for further investigations.

developmental biology↗

Multi-Template Matching: a versatile tool for object-localization in microscopy images

We implemented multiple template matching as both a Fiji plugin and a KNIME workflow, providing an easy-to-use method for the automatic localization of objects of interest in images. We demonstrate its application for the localization of entire or partial biological objects. The Fiji plugin can be installed by activating the Multi-Template-Matching and IJ-OpenCV update sites. The KNIME workflow can be downloaded from nodepit space or the associated GitHub repository. Python source codes and documentations are available on the following GitHub repositories: LauLauThom/MultiTemplateMatching and LauLauThom/MultipleTemplateMatching-KNIME.

bioinformatics↗