Kaiso (ZBTB33) subcellular partitioning functionally links LC3A/B, the tumor microenvironment, and breast cancer survival

The use of digital pathology for the histomorphologic profiling of pathological specimens is expanding the precision and specificity of quantitative tissue analysis at an unprecedented scale; thus, enabling the discovery of new and functionally relevant histological features of both predictive and prognostic significance. In this study, we apply quantitative automated image processing and computational methods to profile the subcellular distribution of the multi-functional transcriptional regulator, Kaiso (ZBTB33), in the tumors of a large racially diverse breast cancer cohort from a designated health disparities region in the United States. Multiplex multivariate analysis of the association of Kaiso’s subcellular distribution with other breast cancer biomarkers reveals novel functional and predictive linkages between Kaiso and the autophagy-related proteins, LC3A/B, that are associated with features of the tumor immune microenvironment, survival, and race. These findings identify effective modalities of Kaiso biomarker assessment and uncover unanticipated insights into Kaiso’s role in breast cancer progression.Fil: Singhal, Sandeep K.. North Dakota State University; Estados UnidosFil: Byun, Jung S.. National Institutes of Health; Estados UnidosFil: Park, Samson. National Institutes of Health; Estados UnidosFil: Yan, Tingfen. National Institutes of Health; Estados UnidosFil: Yancey, Ryan. Columbia University; Estados UnidosFil: Caban, Ambar. Columbia University; Estados UnidosFil: Hernandez, Sara Gil. National Institutes of Health; Estados UnidosFil: Hewitt, Stephen M.. U.S. Department of Health & Human Services. National Institute of Health. National Cancer Institute; Estados UnidosFil: Boisvert, Heike. Ultivue, Inc; Reino UnidoFil: Hennek, Stephanie. Ultivue Inc.; Reino UnidoFil: Bobrow, Mark. Ultivue Inc.; Reino UnidoFil: Ahmed, Md Shakir Uddin. Tuskegee University; Estados UnidosFil: White, Jason. Tuskegee University; Estados UnidosFil: Yates, Clayton. Tuskegee University; Estados UnidosFil: Aukerman, Andrew. Columbia University; Estados UnidosFil: Vanguri, Rami. Columbia University; Estados UnidosFil: Bareja, Rohan. Columbia University; Estados UnidosFil: Lenci, Romina. Columbia University; Estados UnidosFil: Farré, Paula Lucía. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental. Fundación de Instituto de Biología y Medicina Experimental. Instituto de Biología y Medicina Experimental; ArgentinaFil: de Siervi, Adriana. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental. Fundación de Instituto de Biología y Medicina Experimental. Instituto de Biología y Medicina Experimental; ArgentinaFil: Nápoles, Anna María. National Institutes of Health; Estados UnidosFil: Vohra, Nasreen. East Carolina University; Estados UnidosFil: Gardner, Kevin. Columbia University; Estados Unido

Physiological Correlates of Volunteering

We review research on physiological correlates of volunteering, a neglected but promising research field. Some of these correlates seem to be causal factors influencing volunteering. Volunteers tend to have better physical health, both self-reported and expert-assessed, better mental health, and perform better on cognitive tasks. Research thus far has rarely examined neurological, neurochemical, hormonal, and genetic correlates of volunteering to any significant extent, especially controlling for other factors as potential confounds. Evolutionary theory and behavioral genetic research suggest the importance of such physiological factors in humans. Basically, many aspects of social relationships and social activities have effects on health (e.g., Newman and Roberts 2013; Uchino 2004), as the widely used biopsychosocial (BPS) model suggests (Institute of Medicine 2001). Studies of formal volunteering (FV), charitable giving, and altruistic behavior suggest that physiological characteristics are related to volunteering, including specific genes (such as oxytocin receptor [OXTR] genes, Arginine vasopressin receptor [AVPR] genes, dopamine D4 receptor [DRD4] genes, and 5-HTTLPR). We recommend that future research on physiological factors be extended to non-Western populations, focusing specifically on volunteering, and differentiating between different forms and types of volunteering and civic participation

Translocation of Porphyromonas gingivalis Gingipain Adhesin Peptide A44 to Host Mitochondria Prevents Apoptosis ▿

Porphyromonas gingivalis, a Gram-negative oral anaerobe, is associated with periodontal diseases that, in some form, affect up to 80% of the U.S. population. The organism is highly proteolytic, and noncatalytic adhesin domains of the major proteases, gingipains, are involved in bacterium-host interactions. Recently, we showed that gingipain adhesin peptide A44 hijacks the host's clathrin-dependent endocytosis system, allowing the peptide and whole bacteria to be internalized by epithelial cells. In the present study, we found by cell fractionation assays and confocal microscopy that peptide A44 translocated to host mitochondria. Cell viability assays and quantitative real-time PCR showed that the peptide interacted with the cell death machinery by triggering upregulation of antiapoptotic factors bcl-2 and bcl-XL and prevented staurosporine-induced apoptosis for up to 12 h. We confirmed these findings with Western blot analyses of caspase-9 activation in time course experiments with staurosporine. Finally, we verified a similar antiapoptotic effect for P. gingivalis, showing for the first time that the organism manipulated mitochondrial functions during the first hours of infection, thus resisting host cell clearance by apoptosis of infected cells. This mechanism may enable the bacteria to persist in the protected cellular environment until the next step in pathogenesis, progression or resolution of infection

Rga is a regulator of adherence and pilus formation in Streptococcus agalactiae

Streptococcus agalactiae is the leading cause of bacterial sepsis and meningitis in neonates and is also the causative agent of several serious infections in immunocompromised adults. S. agalactiae encounters multiple niches during an infection, suggesting that regulatory mechanisms control the expression of specific virulence factors in this bacterium. The present study describes the functional characterization of a gene from S. agalactiae, designated rga, which encodes a protein with significant similarity to members of the RofA-like protein (RALP) family of transcriptional regulators. After deletion of the rga gene in the genome of S. agalactiae, the mutant strain exhibited significantly reduced expression of the genes srr-1 and pilA, which encode a serine-rich repeat surface glycoprotein and a pilus protein, respectively, and moderately increased expression of the fbsA gene, which encodes a fibrinogen-binding protein. Electrophoretic mobility shift assays demonstrated specific DNA binding of purified Rga to the promoter regions of pilA and fbsA, suggesting that Rga directly controls pilA and fbsA. Adherence assays revealed significantly reduced binding of the Δrga mutant to epithelial HEp-2 cells and to immobilized human keratin 4, respectively. In contrast, the adherence of the Δrga mutant to A549 cells and its binding to human fibrinogen was significantly increased. Immunoblot and immunoelectron microscopy revealed that the quantity of pilus structures was significantly reduced in the Δrga mutant compared with the parental strain. The wild-type phenotype could be restored by plasmid-mediated expression of rga, demonstrating that the mutant phenotypes resulted from a loss of Rga function

Bacteria-derived hydrogen sulfide promotes IL-8 production from epithelial cells

Hydrogen Sulfide (H(2)S) a volatile Sulfur compound, is implicated as a cause of inflammation. especially when it is produced by bacteria colonizing gastrointestinal organs However, It IS Unclear if H(2)S produced by periodontal pathogens affects the inflammatory responses mediated by oral/gingival epithelial cells Therefore. the aims of this Study were (1) to compare the in vitro production of H(2)S among. 14 strains of Oral bacteria and (2) to evaluate the effects of H(2)S on inflammatory response induced in host oral/gingival epithelial cells Porphyromonas gingivalis (Pg) produced the most H(2)S in Culture, Which, in turn resulted in the promotion of proinflammatory cytokine IL-8 from both gingival and Oral epithelial cells The up-regulation of IL-8 expression was reproduced by the exogenously applied H(2)S Furthermore. the Mutant Strains of Pg that do not produce major Soluble Virulent factors. ie gingival, still showed the Production of H(2)S. as well as the promotion of epithelial IL-8 production. which was abrogated by H(2)S scavenging reagents These results demonstrated that Pg produces a concentration of H(2)S capable of Up-regulating-IL-8 expression induced in gingival and oral epithelial cells, revealing a possible mechanism that may promote the inflammation in periodontal disease (C) 2009 Elsevier B.V. All rights reserve

EMSA analysis with promoter regions of genes potentially regulated by VicR and/or CovR.

<p>Genes which showed significant fold-changes in expression in UAvic and/or UAcov mutants compared to UA159 were selected. (A) Recombinant VicR protein (rVicR) specifically bound to the promoter regions of <i>wapE</i>, <i>lysM</i>, <i>smaA</i> and <i>SMU.367</i>; <i>covR</i> was a negative control fragment of similar size. (B) rCovR specifically bound to the promoters of <i>wapE</i>, <i>lysM</i>, <i>epsC</i> and <i>gbpB</i>, but not to <i>gtfD</i>. (C) EMSA assays performed in the presence of the two regulators (rCovR and rVicR) showed co-binding to the promoter regions of <i>lysM</i>, <i>gbpB</i> and <i>gtfC</i>. Co-binding was not observed with the promoter fragments of <i>smaA</i> and <i>epsC</i>, which are exclusively regulated by VicR and CovR, respectively. Specificity of binding was confirmed in competitive assays with excess of cold DNA.</p

Comparison of biomasses of 8 h biofilms formed in the presence of sucrose.

<p>Columns represent means of three independent experiments performed in six replicates. Bars indicate standard deviations. Significant differences compared to parent strain UA159 (dashed column) are indicated by asterisks (* p<0.05; ANOVA with <i>post hoc</i> Dunnett‘s test).</p

Quantitative comparisons of SEM images of 2 h biofilms using Image J software.

<p>(A) Columns represent mean numbers of matrix-based microcolonies, and (B) mean coverage areas (µm²) determined in 32 pre-determined areas per strain in one representative experiment. Bars indicate standard deviations. Asterisks indicate statistically significant differenced compared to parent UA159 (* p<0.05; Kruskal Wallis with <i>post hoc</i> Dunn's multiple comparison).</p

CovR and VicRK Regulate Cell Surface Biogenesis Genes Required for Biofilm Formation in <em>Streptococcus mutans</em>

<div><p>The two-component system VicRK and the orphan regulator CovR of <i>Streptococcus mutans</i> co-regulate a group of virulence genes associated with the synthesis of and interaction with extracellular polysaccharides of the biofilm matrix. Knockout mutants of v<i>icK</i> and <i>covR</i> display abnormal cell division and morphology phenotypes, although the gene function defects involved are as yet unknown. Using transcriptomic comparisons between parent strain UA159 with <i>vicK</i> (UAvic) or <i>covR</i> (UAcov) deletion mutants together with electrophoretic motility shift assays (EMSA), we identified genes directly regulated by both VicR and CovR with putative functions in cell wall/surface biogenesis, including <i>gbpB, wapE, smaA, SMU.2146c</i>, and <i>lysM.</i> Deletion mutants of genes regulated by VicR and CovR (<i>wapE, lysM, smaA</i>), or regulated only by VicR (<i>SMU.2146c</i>) or CovR (<i>epsC</i>) promoted significant alterations in biofilm initiation, including increased fragility, defects in microcolony formation, and atypical cell morphology and/or chaining. Significant reductions in mureinolytic activity and/or increases in DNA release during growth were observed in knockout mutants of <i>smaA</i>, <i>wapE, lysM</i>, SMU.<i>2146c</i> and <i>epsC</i>, implying roles in cell wall biogenesis. <i>WapE</i> and <i>lysM</i> mutations also affected cell hydrophobicity and sensitivity to osmotic or oxidative stress. Finally, <i>vicR, covR</i> and VicRK/CovR-targets (<i>gbpB, wapE, smaA</i>, <i>SMU.2146c, lysM, epsC</i>) are up-regulated in UA159 during biofilm initiation, in a sucrose-dependent manner. These data support a model in which VicRK and CovR coordinate cell division and surface biogenesis with the extracellular synthesis of polysaccharides, a process apparently required for formation of structurally stable biofilms in the presence of sucrose.</p> </div