Mechanisms of Candida albicans Trafficking to the Brain

During hematogenously disseminated disease, Candida albicans infects most organs, including the brain. We discovered that a C. albicans vps51Δ/Δ mutant had significantly increased tropism for the brain in the mouse model of disseminated disease. To investigate the mechanisms of this enhanced trafficking to the brain, we studied the interactions of wild-type C. albicans and the vps51Δ/Δ mutant with brain microvascular endothelial cells in vitro. These studies revealed that C. albicans invasion of brain endothelial cells is mediated by the fungal invasins, Als3 and Ssa1. Als3 binds to the gp96 heat shock protein, which is expressed on the surface of brain endothelial cells, but not human umbilical vein endothelial cells, whereas Ssa1 binds to a brain endothelial cell receptor other than gp96. The vps51Δ/Δ mutant has increased surface expression of Als3, which is a major cause of the increased capacity of this mutant to both invade brain endothelial cells in vitro and traffic to the brain in mice. Therefore, during disseminated disease, C. albicans traffics to and infects the brain by binding to gp96, a unique receptor that is expressed specifically on the surface of brain endothelial cells

SR-Like RNA-binding protein Slr1 affects Candida albicans filamentation and virulence

Candida albicans causes both mucosal and disseminated infections, and its capacity to grow as both yeast and hyphae is a key virulence factor. Hyphal formation is a type of polarized growth, and members of the SR (serine-arginine) family of RNA-binding proteins influence polarized growth of both Saccharomyces cerevisiae and Aspergillus nidulans. Therefore, we investigated whether SR-like proteins affect filamentous growth and virulence of C. albicans. BLAST searches with S. cerevisiae SR-like protein Npl3 (ScNpl3) identified two C. albicans proteins: CaNpl3, an apparent ScNpl3 ortholog, and Slr1, another SR-like RNAbinding protein with no close S. cerevisiae ortholog. Whereas ScNpl3 was critical for growth, deletion of NPL3 in C. albicans resulted in few phenotypic changes. In contrast, the slr1δ/δ mutant had a reduced growth rate in vitro, decreased filamentation, and impaired capacity to damage epithelial and endothelial cells in vitro. Mice infected intravenously with the slr1δ/δ mutant strain had significantly prolonged survival compared to that of mice infected with the wild-type or slr1δ/δ mutant complemented with SLR1 (slr1δ/δ+SLR1) strain, without a concomitant decrease in kidney fungal burden. Histopathology, however, revealed differential localization of slr1δ/δ hyphal and yeast morphologies within the kidney. Mice infected with slr1δ/δ cells also had an increased brain fungal burden, which correlated with increased invasion of brain, but not umbilical vein, endothelial cells in vitro. The enhanced brain endothelial cell invasion was likely due to the increased surface exposure of the Als3 adhesin on slr1δ/δ cells. Our results indicate that Slr1 is an SR-like protein that influences C. albicans growth, filamentation, host cell interactions, and virulence. © 2013, American Society for Microbiology

Highly controllable and reliable ultra-thin Parylene deposition

Thanks to the excellent barrier property and fabrication accessibility, Parylene has been actively used in the microelectromechanical system. An ultra-thin Parylene film with thickness smaller than 100 nm is usually required to precisely tune the surface property of substrate or protect the functional unit. The commercially available regular Parylene deposition is a dimer mass determined chemical vapor deposition process with a high output (i.e. a low deposition precision in term of thickness control), around 1.6 μm/g (the ratio of film thickness to the loaded dimer mass) for the machine in the author’s lab. Therefore, it is hard to controllably and reliably prepare a Parylene film with thickness smaller than 100 nm, which requires a dimer mass less than 62.5 mg. This paper reported a method to prepare ultra-thin Parylene films with the nominal thickness down to 1 nm. A home-made deposition chamber was put inside and connected with the regular machine chamber through a microfabricated orifice with feature size smaller than 1 mm. According to the free molecular flow theory, the pressure inside the deposition chamber can be predictably and controllably reduced, thereby an ultra-low output of Parylene deposition, as low as 0.08 nm/g, was successfully obtained. The deposition precision was increased by 4 orders of magnitude compared to that of a direct Parylene deposition. This highly controllable and reliable ultra-thin Parylene deposition technique will find promising applications in flexible electronics and biomedical microdevices

Antisense RNA associated with biological regulation of a restriction–modification system

Restriction–modification systems consist of a modification enzyme that methylates a specific DNA sequence and a restriction endonuclease that cleaves DNA lacking this epigenetic signature. Their gene expression should be finely regulated because their potential to attack the host bacterial genome needs to be controlled. In the EcoRI system, where the restriction gene is located upstream of the modification gene in the same orientation, we previously identified intragenic reverse promoters affecting gene expression. In the present work, we identified a small (88 nt) antisense RNA (Rna0) transcribed from a reverse promoter (PREV0) at the 3′ end of the restriction gene. Its antisense transcription, as measured by transcriptional gene fusion, appeared to be terminated by the PM1,M2 promoter. PM1,M2 promoter-initiated transcription, in turn, appeared to be inhibited by PREV0. Mutational inactivation of PREV0 increased expression of the restriction gene. The biological significance of this antisense transcription is 2-fold. First, a mutation in PREV0 increased restriction of incoming DNA. Second, the presence of the antisense RNA gene (ecoRIA) in trans alleviated cell killing after loss of the EcoRI plasmid (post-segregational killing). Taken together, these results strongly suggested the involvement of an antisense RNA in the biological regulation of this restriction–modification system

Prevalence and Determinants of Metabolic Syndrome among Women in Chinese Rural Areas

BACKGROUND AND AIMS: Metabolic syndrome (MS) is prevalent in recent years but few data is reported in the rural areas in China. The aim of this study was to examine MS prevalence and its risk factors among women in rural China. METHODS AND RESULTS: The Nantong Metabolic Syndrome Study (NMSS), a population based cross-sectional study, was conducted during 2007-2008 in Nantong, China. In person interviews, blood glucose and lipid measurements were completed for 13,505 female participants aged 18-74 years. The International Diabetes Federation (IDF), the US Third Report of the National Cholesterol Education Program, the Adult Treatment Panel (ATPIII) and modified ATPIII for Asian population has determined three criteria of MS. These criteria for MS were used and compared in this study. The prevalence of MS was 22.0%, 16.9% and 23.3% according to IDF, ATPIII and ATPIII-modified criteria, respectively. Levels of agreement of these criteria for MS were above 0.75. We found that vigorous-intensity of occupational physical activity was associated with a low prevalence of MS with OR of 0.76 (95% confidence interval (CI): 0.63-0.91). Rice wine drinkers (alcohol >12.8 g/day) had about 34% low risks of developing MS with OR of 0.66 (95% CI: 0.48-0.91), compared with non-drinkers. Odds ratio of MS was 1.81 (95% CI: 1.15-2.84) in women who smoked more than 20 pack-years, compared to non-smokers. Odds ratio of MS was 1.56 (95% CI: 1.25-1.95) in women who had familial history of diseases, including hypertension, diabetes and stroke, compared to women without familial history of those diseases. CONCLUSION: MS is highly prevalent among women in rural China. Both physical activity and rice wine consumption play a protective role, while family history and smoking are risk factors in MS development. Educational programs should be established for promoting healthy lifestyles and appropriate interventions in rural China

Genomic Analyses Reveal Mutational Signatures and Frequently Altered Genes in Esophageal Squamous Cell Carcinoma

Esophageal squamous cell carcinoma (ESCC) is one of the most common cancers worldwide and the fourth most lethal cancer in China. However, although genomic studies have identified some mutations associated with ESCC, we know little of the mutational processes responsible. To identify genome-wide mutational signatures, we performed either whole-genome sequencing (WGS) or whole-exome sequencing (WES) on 104 ESCC individuals and combined our data with those of 88 previously reported samples. An APOBEC-mediated mutational signature in 47% of 192 tumors suggests that APOBEC-catalyzed deamination provides a source of DNA damage in ESCC. Moreover, PIK3CA hotspot mutations (c.1624G>A [p.Glu542Lys] and c.1633G>A [p.Glu545Lys]) were enriched in APOBEC-signature tumors, and no smoking-associated signature was observed in ESCC. In the samples analyzed by WGS, we identified focal (<100 kb) amplifications of CBX4 and CBX8. In our combined cohort, we identified frequent inactivating mutations in AJUBA, ZNF750, and PTCH1 and the chromatin-remodeling genes CREBBP and BAP1, in addition to known mutations. Functional analyses suggest roles for several genes (CBX4, CBX8, AJUBA, and ZNF750) in ESCC. Notably, high activity of hedgehog signaling and the PI3K pathway in approximately 60% of 104 ESCC tumors indicates that therapies targeting these pathways might be particularly promising strategies for ESCC. Collectively, our data provide comprehensive insights into the mutational signatures of ESCC and identify markers for early diagnosis and potential therapeutic targets

Requirement of argininosuccinate lyase for systemic nitric oxide production

Nitric oxide (NO) is crucial in diverse physiological and pathological processes. We show that a hypomorphic mouse model of argininosuccinate lyase (encoded by Asl) deficiency has a distinct phenotype of multiorgan dysfunction and NO deficiency. Loss of Asl in both humans and mice leads to reduced NO synthesis, owing to both decreased endogenous arginine synthesis and an impaired ability to use extracellular arginine for NO production. Administration of nitrite, which can be converted into NO in vivo, rescued the manifestations of NO deficiency in hypomorphic Asl mice, and a nitric oxide synthase (NOS)-independent NO donor restored NO-dependent vascular reactivity in humans with ASL deficiency. Mechanistic studies showed that ASL has a structural function in addition to its catalytic activity, by which it contributes to the formation of a multiprotein complex required for NO production. Our data demonstrate a previously unappreciated role for ASL in NOS function and NO homeostasis. Hence, ASL may serve as a target for manipulating NO production in experimental models, as well as for the treatment of NO-related diseases

Host Cell Invasion and Virulence Mediated by Candida albicans Ssa1

Candida albicans Ssa1 and Ssa2 are members of the HSP70 family of heat shock proteins that are expressed on the cell surface and function as receptors for antimicrobial peptides such as histatins. We investigated the role of Ssa1 and Ssa2 in mediating pathogenic host cell interactions and virulence. A C. albicans ssa1Δ/Δ mutant had attenuated virulence in murine models of disseminated and oropharyngeal candidiasis, whereas an ssa2Δ/Δ mutant did not. In vitro studies revealed that the ssa1Δ/Δ mutant caused markedly less damage to endothelial cells and oral epithelial cell lines. Also, the ssa1Δ/Δ mutant had defective binding to endothelial cell N-cadherin and epithelial cell E-cadherin, receptors that mediate host cell endocytosis of C. albicans. As a result, this mutant had impaired capacity to induce its own endocytosis by endothelial cells and oral epithelial cells. Latex beads coated with recombinant Ssa1 were avidly endocytosed by both endothelial cells and oral epithelial cells, demonstrating that Ssa1 is sufficient to induce host cell endocytosis. These results indicate that Ssa1 is a novel invasin that binds to host cell cadherins, induces host cell endocytosis, and is critical for C. albicans to cause maximal damage to host cells and induce disseminated and oropharyngeal disease