Adenylyl Cyclase Plays a Regulatory Role in Development, Stress Resistance and Secondary Metabolism in Fusarium fujikuroi

The ascomycete fungus Fusarium fujikuroi (Gibberella fujikuroi MP-C) produces secondary metabolites of biotechnological interest, such as gibberellins, bikaverin, and carotenoids. Production of these metabolites is regulated by nitrogen availability and, in a specific manner, by other environmental signals, such as light in the case of the carotenoid pathway. A complex regulatory network controlling these processes is recently emerging from the alterations of metabolite production found through the mutation of different regulatory genes. Here we show the effect of the targeted mutation of the acyA gene of F. fujikuroi, coding for adenylyl cyclase. Mutants lacking the catalytic domain of the AcyA protein showed different phenotypic alterations, including reduced growth, enhanced production of unidentified red pigments, reduced production of gibberellins and partially derepressed carotenoid biosynthesis in the dark. The phenotype differs in some aspects from that of similar mutants of the close relatives F. proliferatum and F. verticillioides: contrary to what was observed in these species, ΔacyA mutants of F. fujikuroi showed enhanced sensitivity to oxidative stress (H2O2), but no change in heavy metal resistance or in the ability to colonize tomato tissue, indicating a high versatility in the regulatory roles played by cAMP in this fungal group

TWEAK Affects Keratinocyte G2/M Growth Arrest and Induces Apoptosis through the Translocation of the AIF Protein to the Nucleus

The soluble TNF-like weak inducer of apoptosis (TWEAK, TNFSF12) binds to the fibroblast growth factor-inducible 14 receptor (FN14, TNFRSF12A) on the cell membrane and induces multiple biological responses, such as proliferation, migration, differentiation, angiogenesis and apoptosis. Previous reports show that TWEAK, which does not contain a death domain in its cytoplasmic tail, induces the apoptosis of tumor cell lines through the induction of TNFα secretion. TWEAK induces apoptosis in human keratinocytes. Our experiments clearly demonstrate that TWEAK does not induce the secretion of TNFα or TRAIL proteins. The use of specific inhibitors and the absence of procaspase-3 cleavage suggest that the apoptosis of keratinocytes follows a caspase- and cathepsin B-independent pathway. Further investigation showed that TWEAK induces a decrease in the mitochondrial membrane potential of keratinocytes. Confocal microscopy showed that TWEAK induces the cleavage and the translocation of apoptosis inducing factor (AIF) from the mitochondria to the nucleus, thus initiating caspase-independent apoptosis. Moreover, TWEAK induces FOXO3 and GADD45 expression, cdc2 phosphorylation and cdc2 and cyclinB1 degradation, resulting in the arrest of cell growth at the G2/M phase. Finally, we report that TWEAK and FN14 are normally expressed in the basal layer of the physiological epidermis and are greatly enhanced in benign (psoriasis) and malignant (squamous cell carcinoma) skin pathologies that are characterized by an inflammatory component. TWEAK might play an essential role in skin homeostasis and pathology

The Stress Response Factors Yap6, Cin5, Phd1, and Skn7 Direct Targeting of the Conserved Co-Repressor Tup1-Ssn6 in S. cerevisiae

Maintaining the proper expression of the transcriptome during development or in response to a changing environment requires a delicate balance between transcriptional regulators with activating and repressing functions. The budding yeast transcriptional co-repressor Tup1-Ssn6 is a model for studying similar repressor complexes in multicellular eukaryotes. Tup1-Ssn6 does not bind DNA directly, but is directed to individual promoters by one or more DNA-binding proteins, referred to as Tup1 recruiters. This functional architecture allows the Tup1-Ssn6 to modulate the expression of genes required for the response to a variety of cellular stresses. To understand the targeting or the Tup1-Ssn6 complex, we determined the genomic distribution of Tup1 and Ssn6 by ChIP-chip. We found that most loci bound by Tup1-Ssn6 could not be explained by co-occupancy with a known recruiting cofactor and that deletion of individual known Tup1 recruiters did not significantly alter the Tup1 binding profile. These observations suggest that new Tup1 recruiting proteins remain to be discovered and that Tup1 recruitment typically depends on multiple recruiting cofactors. To identify new recruiting proteins, we computationally screened for factors with binding patterns similar to the observed Tup1-Ssn6 genomic distribution. Four top candidates, Cin5, Skn7, Phd1, and Yap6, all known to be associated with stress response gene regulation, were experimentally confirmed to physically interact with Tup1 and/or Ssn6. Incorporating these new recruitment cofactors with previously characterized cofactors now explains the majority of Tup1 targeting across the genome, and expands our understanding of the mechanism by which Tup1-Ssn6 is directed to its targets

Race-Based Traffic-Stops: "Do They Really Occur?"

Investigates the occurrence of race-based traffic stops and finds that they do exist and recommends that department gather data in order to assess the situation accurately

Antiviral Coatings as Continuously Active Disinfectants

Antimicrobial surfaces and coatings have been available for many decades and have largely been designed to kill or prevent the growth of bacteria and fungi. Antiviral coatings have become of particular interest more recently during the COVID-19 pandemic as they are designed to act as continuously active disinfectants. The most studied antiviral coatings have been metal-based or are comprised of silane quaternary ammonium formulations. Copper and silver interact directly with proteins and nucleic acids, and influence the production of reactive free radicals. Titanium dioxide acts as a photocatalyst in the presence of water and oxygen to produce free radicals in the presence of UV light or visible light when alloyed with copper or silver. Silane quaternary ammonium formulations can be applied to surfaces using sprays or wipes, and are particularly effective against enveloped viruses. Continuously active disinfectants offer an extra barrier against fomite-mediated transmission of respiratory and enteric viruses to reduce exposure between routine disinfection and cleaning events. To take advantage of this technology, testing methods need to be standardized and the benefits quantified in terms of reduction of virus transmission

Inactivation kinetics of benzalkonium chloride and ethanol-based hand sanitizers against a betacoronavirus and an alphacoronavirus

Summary: Background: Hand hygiene is critical to lower the potential for the spread of SARS-CoV-2 and other infectious agents by direct contact. When running water and soap are not available for hand hygiene, ethanol-based hand sanitizers are currently the recommended standard of care [1–3]. Though recently published data showed comparable in vitro effectiveness of benzalkonium chloride (BAK)-based and ethanol-based hand sanitizers against SARS-CoV-2 virus, a paucity of peer-reviewed data on the effectiveness of these formulations against other types of infective coronaviruses remains. This work assessed human coronavirus HCoV-229E (genus Alphacoronavirus) concurrently with SARS-CoV-2, Isolate USA-WA1/2020 (genus Betacoronavirus) to fill this gap. Methods: The test was conducted according to EN14476:2013-A2:2019 [EN14476] Quantitative Suspension Test for the Evaluation of Virucidal Activity in the Medical Area [4]. Two BAK-based hand sanitizers, five ethanol-based hand sanitizers, and an 80% ethanol reference formulation were tested for antiviral activity against SARS-CoV-2 and HCoV-229E at 15- and 30- second contact times. Results: Both SARS-CoV-2 and HCoV-229E were reduced by greater than 4.00-log10 within 15 seconds of contact. Virus decay constants (k) following first-order kinetics were similar for BAK and ethanol-based formulations against both test viruses. The SARS-CoV-2 results reported herein mirrored previous data reported by Herdt et al. (2021). Conclusion: BAK and ethanol hand sanitizer formulations inactivate SARS-CoV-2 and HCoV-229E at similar rates. This data supports previously published effectiveness data for both chemistries and indicates that additional coronavirus strains and variants would demonstrate similar inactivation trends

Different intensity extension methods and their impact on entrance dose in breast radiotherapy: A study

In breast radiotherapy, skin flashing of treatment fields is important to account for intrafraction movements and setup errors. This study compares the two different intensity extension methods, namely, Virtual Bolus method and skin flash tool method, to provide skin flashing in intensity modulated treatment fields. The impact of these two different intensity extension methods on skin dose was studied by measuring the entrance dose of the treatment fields using semiconductor diode detectors. We found no significant difference in entrance dose due to different methods used for intensity extension. However, in the skin flash tool method, selection of appropriate parameters is important to get optimum fluence extension