UDP-glucose 4, 6-dehydratase Activity Plays an Important Role in Maintaining Cell Wall Integrity and Virulence of Candida albicans

Candida albicans, a human fungal pathogen, undergoes morphogenetic changes that are associated with virulence. We report here that GAL102 in C. albicans encodes a homolog of dTDP-glucose 4,6-dehydratase, an enzyme that affects cell wall properties as well as virulence of many pathogenic bacteria. We found that GAL102 deletion leads to greater sensitivity to antifungal drugs and cell wall destabilizing agents like Calcofluor white and Congo red. The mutant also formed biofilms consisting mainly of hyphal cells that show less turgor. The NMR analysis of cell wall mannans of gal102 deletion strain revealed that a major constituent of mannan is missing and the phosphomannan component known to affect virulence is greatly reduced. We also observed that there was a substantial reduction in the expression of genes involved in biofilm formation but increase in the expression of genes encoding glycosylphosphatidylinositol-anchored proteins in the mutant. These, along with altered mannosylation of cell wall proteins together might be responsible for multiple phenotypes displayed by the mutant. Finally, the mutant was unable to grow in the presence of resident peritoneal macrophages and elicited a weak pro-inflammatory cytokine response in vitro. Similarly, this mutant elicited a poor serum pro-inflammatory cytokine response as judged by IFNγ and TNFα levels and showed reduced virulence in a mouse model of systemic candidiasis. Importantly, an Ala substitution for a conserved Lys residue in the active site motif YXXXK, that abrogates the enzyme activity also showed reduced virulence and increased filamentation similar to the gal102 deletion strain. Since inactivating the enzyme encoded by GAL102 makes the cells sensitive to antifungal drugs and reduces its virulence, it can serve as a potential drug target in combination therapies for C. albicans and related pathogens

Entry of Human Papillomavirus Type 16 by Actin-Dependent, Clathrin- and Lipid Raft-Independent Endocytosis

Infectious endocytosis of incoming human papillomavirus type 16 (HPV-16), the main etiological agent of cervical cancer, is poorly characterized in terms of cellular requirements and pathways. Conflicting reports attribute HPV-16 entry to clathrin-dependent and -independent mechanisms. To comprehensively describe the cell biological features of HPV-16 entry into human epithelial cells, we compared HPV-16 pseudovirion (PsV) infection in the context of cell perturbations (drug inhibition, siRNA silencing, overexpression of dominant mutants) to five other viruses (influenza A virus, Semliki Forest virus, simian virus 40, vesicular stomatitis virus, and vaccinia virus) with defined endocytic requirements. Our analysis included infection data, i.e. GFP expression after plasmid delivery by HPV-16 PsV, and endocytosis assays in combination with electron, immunofluorescence, and video microscopy. The results indicated that HPV-16 entry into HeLa and HaCaT cells was clathrin-, caveolin-, cholesterol- and dynamin-independent. The virus made use of a potentially novel ligand-induced endocytic pathway related to macropinocytosis. This pathway was distinct from classical macropinocytosis in regards to vesicle size, cholesterol-sensitivity, and GTPase requirements, but similar in respect to the need for tyrosine kinase signaling, actin dynamics, Na+/H+ exchangers, PAK-1 and PKC. After internalization the virus was transported to late endosomes and/or endolysosomes, and activated through exposure to low pH

Building Resilience to Climate Change: Pilot Evaluation of the Impact of India’s First Heat Action Plan on All-Cause Mortality

Background. Ahmedabad implemented South Asia’s first heat action plan (HAP) after a 2010 heatwave. This study evaluates the HAP’s impact on all-cause mortality in 2014–2015 relative to a 2007–2010 baseline. Methods. We analyzed daily maximum temperature (Tmax)-mortality relationships before and after HAP. We estimated rate ratios (RRs) for daily mortality using distributed lag nonlinear models and mortality incidence rates (IRs) for HAP warning days, comparing pre- and post-HAP periods, and calculated incidence rate ratios (IRRs). We estimated the number of deaths avoided after HAP implementation using pre- and post-HAP IRs. Results. The maximum pre-HAP RR was 2.34 (95%CI 1.98–2.76) at 47°C (lag 0), and the maximum post-HAP RR was 1.25 (1.02–1.53) estimated at 47°C (lag 0). Post-to-pre-HAP nonlagged mortality IRR for Tmax over 40°C was 0.95 (0.73–1.22) and 0.73 (0.29–1.81) for Tmax over 45°C. An estimated 1,190 (95%CI 162–2,218) average annualized deaths were avoided in the post-HAP period. Conclusion. Extreme heat and HAP warnings after implementation were associated with decreased summertime all-cause mortality rates, with largest declines at highest temperatures. Ahmedabad’s plan can serve as a guide for other cities attempting to increase resilience to extreme heat