Treatment of calcific arterial disease via enhancement of autophagy using GSK343.

Vascular calcification is a hallmark of atherosclerotic disease and serves as a strong predictor and risk factor for cardiovascular events. Growing evidence suggests that autophagy may play a protective role in early atherosclerosis. The precise effects of autophagy on VSMC-mediated calcification remain unknown. In this study, we utilized multi-omic profiling to investigate impaired autophagy at the transcriptional level as a key driver of VSMC calcification. Our findings revealed that impaired autophagy is an essential determinant of VSMC calcification. We observed that an osteogenic environment affects the open chromatin status of VSMCs, compromising the transcriptional activation of autophagy initiation genes. In vivo experiments involve pharmacological and genetic activation of autophagy using mouse models of spontaneous large (Mgp-/-) and small (Abcc6-/-) artery calcification. Taken together, these data advance our mechanistic understanding of vascular calcification and provide important insights for a broad range of cardiovascular diseases involving VSMC phenotype switch.

Prophylactic Surgery for Thoracic Aortic Aneurysm: Asking Family for Help.

A 68-year-old man presented for evaluation of a 5.0-cm asymptomatic ascending aortic aneurysm. He was managed medically until a family member was also diagnosed with a thoracic aortic aneurysm. His aneurysm was, therefore, likely to be genetically predisposed, and he underwent early prophylactic ascending aorta replacement. (Level of Difficulty: Beginner.).

Fluorescence Detection of Type III Secretion Using a Glu-CyFur Reporter System in Citrobacter rodentium.

Enteropathogenic Escherichia coli (EPEC) is a major cause of infantile diarrhea worldwide. EPEC and the closely related murine model of EPEC infection, Citrobacter rodentium, utilize a type III secretion system (T3SS) to propagate the infection. Since the T3SS is not essential for the bacteria to survive or propagate, inhibiting the virulence factor with a therapeutic would treat the infection without causing harm to commensal bacteria. Studying inhibitors of the T3SS usually requires a BSL-2 laboratory designation and eukaryotic host cells while not indicating the mechanism of inhibition. We have designed a BSL-1 assay using the murine model C. rodentium that does not require mammalian cell culture. This CPG2-reporter assay allows for more rapid analysis of secretion efficiency than Western blotting and is sensitive enough to differentiate between partial and total inhibition of the T3SS. Here we present our method and the results of a small collection of compounds we have screened, including known T3SS inhibitors EGCG, regacin, and aurodox and related quorum sensing inhibitors tannic acid and ellagic acid. We have further characterized EGCG as a T3SS inhibitor and established its IC50 of 1.8 ± 0.4 µM. We also establish tannic acid as a potent inhibitor of the T3SS with an IC50 of 0.65 ± 0.09 µM.

Recurrent spontaneous pneumothoraces and vaping in an 18-year-old man: a case report and review of the literature.

BACKGROUND: Primary spontaneous pneumothorax is a common disorder occurring in young adults without underlying lung disease. Although tobacco smoking is a well-documented risk factor for spontaneous pneumothorax, an association between electronic cigarette use (that is, vaping) and spontaneous pneumothorax has not been noted. We report a case of spontaneous pneumothoraces correlated with vaping. CASE PRESENTATION: An 18-year-old Caucasian man presented twice with recurrent right-sided spontaneous pneumothoraces within 2 weeks. He reported a history of vaping just prior to both episodes. Diagnostic testing was notable for a right-sided spontaneous pneumothorax on chest X-ray and computed tomography scan. His symptoms improved following insertion of a chest tube and drainage of air on each occasion. In the 2-week follow-up visit for the recurrent episode, he was asymptomatic and reported that he was no longer using electronic cigarettes. CONCLUSIONS: Providers and patients should be aware of the potential risk of spontaneous pneumothorax associated with electronic cigarettes.

Structural modeling and functional analysis of the essential ribosomal processing protease Prp from Staphylococcus aureus.

In Firmicutes and related bacteria, ribosomal large subunit protein L27 is encoded with a conserved N-terminal extension that is removed to expose residues critical for ribosome function. Bacteria encoding L27 with this N-terminal extension also encode a sequence-specific cysteine protease, Prp, which carries out this cleavage. In this work, we demonstrate that L27 variants with an un-cleavable N-terminal extension, or lacking the extension (pre-cleaved), are unable to complement an L27 deletion in Staphylococcus aureus. This indicates that N-terminal processing of L27 is not only essential but possibly has a regulatory role. Prp represents a new clade of previously uncharacterized cysteine proteases, and the dependence of S. aureus on L27 cleavage by Prp validates the enzyme as a target for potential antibiotic development. To better understand the mechanism of Prp activity, we analyzed Prp enzyme kinetics and substrate preference using a fluorogenic peptide cleavage assay. Molecular modeling and site-directed mutagenesis implicate several residues around the active site in catalysis and substrate binding, and support a structural model in which rearrangement of a flexible loop upon binding of the correct peptide substrate is required for the active site to assume the proper conformation. These findings lay the foundation for the development of antimicrobials that target this novel, essential pathway.

Testing intra-hemocelic injection of antimicrobials against Encephalitozoon sp. (Microsporidia) in an insect host.

Encephalitozoon spp. are the primary microsporidial pathogens of humans and domesticated animals. In this experiment, we test the efficacy of four commercial antimicrobials against an Encephalitozoon sp. in an insect host by intra-hemocelic injection. All four antimicrobials, viz., thiabendazole, quinine, albendazole, and fumagillin, significantly reduced but did not eliminate microsporidia spore counts in the grasshopper host. Among these four drugs, thiabendazole was most effective in reducing the microsporidia spore level up to 90%, followed by quinine (70%), albendazole (62%), and fumagillin (59%). No control or quinine-treated animals died, whereas 45% of albendazole animals died. Despite the high mortality induced by albendazole, this drug significantly reduced spore counts, a result not seen in previous per os trials. Among the treatment groups, grasshoppers injected with thiabendazole lost a significant mass. Our study suggests that quinine and related alkaloids should be further examined for antimicrosporidial activity.

Explaining potential antecedents of workplace social support: reciprocity or attractiveness?

Effects of social support are an important topic in occupational stress theories and research, yet little is known about support's potential antecedents. Based on reciprocity theory, the authors hypothesized that the social support received is related to the extent the employee performs organizational citizenship behaviors directed at individuals and to one's social competence; based on the notion of personal attraction, the authors hypothesized that employees' physical attractiveness and sense of humor would be associated with the amount of social support received. In a survey of 123 high school employees and separate ratings of their attractiveness, reciprocity variables were related but attraction variables were not related to social support availability. Further research should examine reciprocity in predicting social support.