Highly sensitive quantitative PCR for the detection and differentiation of Pseudogymnoascus destructans and other Pseudogymnoascus species

White-nose syndrome is a fungal disease that has decimated bat populations across eastern North America. Identification of the etiologic agent, Pseudogymnoascus destructans (formerly Geomyces destructans), in environmental samples is essential to proposed management plans. A major challenge is the presence of closely related species, which are ubiquitous in many soils and cave sediments and often present in high abundance. We present a dual-probe real-time quantitative PCR assay capable of detecting and differentiating P. destructans from closely related fungi in environmental samples from North America. The assay, based on a single nucleotide polymorphism (SNP) specific to P. destructans, is capable of rapid low-level detection from various sampling media, including sediment, fecal samples, wing biopsy specimens, and skin swabs. This method is a highly sensitive, high-throughput method for identifying P. destructans, other Pseudogymnoascus spp., and Geomyces spp. in the environment, providing a fundamental component of research and risk assessment for addressing this disease, as well as other ecological and mycological work on related fungi

Using Mendelian randomisation to identify opportunities for type 2 diabetes prevention by repurposing medications used for lipid management

Background: Maintaining a healthy lifestyle to reduce type 2 diabetes (T2D) risk is challenging and additional strategies for T2D prevention are needed. We evaluated several lipid control medications as potential therapeutic options for T2D prevention using tissue-specific predicted gene expression summary statistics in a two-sample Mendelian randomisation (MR) design. Methods: Large-scale European genome-wide summary statistics for lipids and T2D were leveraged in our multi-stage analysis to estimate changes in either lipid levels or T2D risk driven by tissue-specific predicted gene expression. We incorporated tissue-specific predicted gene expression summary statistics to proxy therapeutic effects of three lipid control medications [i.e., statins, icosapent ethyl (IPE), and proprotein convertase subtilisin/kexin type-9 inhibitors (PCSK-9i)] on T2D susceptibility using two-sample Mendelian randomisation (MR). Findings: IPE, as proxied via increased FADS1 expression, was predicted to lower triglycerides and was associated with a 53% reduced risk of T2D. Statins and PCSK-9i, as proxied by reduced HMGCR and PCSK9 expression, respectively, were predicted to lower LDL-C levels but were not associated with T2D susceptibility. Interpretation: Triglyceride lowering via IPE may reduce the risk of developing T2D in populations of European ancestry. However, experimental validation using animal models is needed to substantiate our results and to motivate randomized control trials (RCTs) for IPE as putative treatment for T2D prevention. Funding: Only summary statistics were used in this analysis. Funding information is detailed under Acknowledgments. © 2022Open access journalThis item from the UA Faculty Publications collection is made available by the University of Arizona with support from the University of Arizona Libraries. If you have questions, please contact us at [email protected]

Does fungicide run-off from citrus delay leaf litter decomposition?

Leaf litter is a major inoculum source for citrus diseases such citrus black spot caused by Phyllosticta citricarpa, and greasy spot caused by Mycosphaerella citri. In order to reduce this inoculum source, the efficacy of urea, dolomitic lime, a commercial compost accelerator, and an organic mulch, was assessed for enhanced leaf decomposition and reduction in sporocarps. However, due to the potential for run-off from high volume fungicide applications to disrupt leaf decomposition and microbial antagonism, the amendments were compared with and without simulated fungicide run-off. Mature green leaves of Citrus sinensis were removed from trees and placed inside mesh bags before being pinned to the orchard floor. The amendments were applied, and then simulated run-off from a typical citrus black spot fungicide program (copper, mancozeb, azoxystrobin) was applied. Leaf degradation was assessed every 2-3 weeks by visual ratings and dry weight. No direct effects on sporocarps could be observed due to insufficient infection. The results showed that the organic mulch was the most effective at enhancing decomposition, while there was significantly (P < 0.05) less decomposition in the presence of fungicide run-off

Identification of resistance to citrus black spot using a novel in-field inoculation assay

Citrus black spot is an important fungal disease of citrus resulting in fruit drop and rind blemish in tropical and subtropical production areas. The disease is incited by the fungus Phyllosticta citricarpa (McAlpine) van der Aa (synonym: Guignardia citricarpa Kiely), with control currently relying on the application of fungicides. Because the presence and expression of resistance is poorly understood, we sought to develop a method for inoculating fruit in the field that gives reproducible symptoms of citrus black spot consistent with natural field infection. We subsequently validated this method by screening 49 citrus accessions and characterized their qualitative expression of citrus black spot symptoms. Challenge inoculations were undertaken with a known isolate of P. citricarpa, and control fruit were inoculated with water or the endophyte P. paracapitalensis Guarnaccia & Crous. Our results showed that all mandarin, sweet orange, lemon and papeda types were susceptible; pummelo, lime, and sour orange types expressed immunity; while various hybrids were susceptible, resistant and immune. Hybrid progeny from crosses using pummelo [Citrus maxima (Burm.) Merr.] as a parent showed preliminary evidence of segregation for citrus black spot immunity. The implications of these results to achieve genetic improvement for citrus black spot resistance in citrus breeding programs are discussed

Baseline characteristics of whites from the discovery cohort stratified by perioperative baseline eGFR.

<p>Baseline characteristics of whites from the discovery cohort stratified by perioperative baseline eGFR.</p

Multivariable logistic regression model for acute kidney injury (AKI) in the discovery cohort adjusted for age, sex, race, body mass index, baseline estimated glomerular filtration rate, history of diabetes, and cardiopulmonary bypass graft use.

<p>Multivariable logistic regression model for acute kidney injury (AKI) in the discovery cohort adjusted for age, sex, race, body mass index, baseline estimated glomerular filtration rate, history of diabetes, and cardiopulmonary bypass graft use.</p

Multivariable logistic regression model for acute kidney injury (AKI) in whites with baseline eGFR > 60 mL/min/1.73m² in the replication cohort adjusted for age, sex, body mass index, baseline estimated glomerular filtration rate, history of diabetes, and cardiopulmonary bypass graft use.

<p>Multivariable logistic regression model for acute kidney injury (AKI) in whites with baseline eGFR > 60 mL/min/1.73m² in the replication cohort adjusted for age, sex, body mass index, baseline estimated glomerular filtration rate, history of diabetes, and cardiopulmonary bypass graft use.</p

Association of gain-of-function <i>EPHX2</i> polymorphism Lys55Arg with acute kidney injury following cardiac surgery

<div><p>Twenty to thirty percent of patients undergoing cardiac surgery develop acute kidney injury (AKI). In mice, inhibition of soluble epoxide hydrolase (sEH) attenuates renal injury following ischemia-reperfusion. We tested the hypothesis that functional variants of <i>EPHX2</i>, encoding sEH, are associated with AKI after cardiac surgery. We genotyped patients in two independent cardiac surgery cohorts for functional <i>EPHX2</i> polymorphisms, Lys55Arg and Arg287Gln, and determined AKI using Acute Kidney Injury Network criteria. The 287Gln variant was not associated with AKI. In the discovery cohort, the gain-of-function 55Arg variant was associated with an increased incidence of AKI in univariate (p = 0.03) and multivariable (p = 0.04) analyses. In white patients without chronic kidney disease (CKD), the 55Arg variant was independently associated with AKI with an OR of 2.04 (95% CI 0.95–4.42) for 55Arg heterozygotes and 31.53 (1.57–633.19) for homozygotes (p = 0.02), after controlling for age, sex, body mass index, baseline estimated glomerular filtration rate, and use of cardiopulmonary bypass. These findings were replicated in the second cardiac surgery cohort. 12,13- and total- dihydroxyoctadecanoic acids (DiHOME): epoxyoctadecanoic acids (EpOME) ratios were increased in <i>EPHX2</i> 55Arg variant carriers, consistent with increased hydrolase activity. The <i>EPHX2</i> Lys55Arg polymorphism is associated with AKI following cardiac surgery in patients without preexisting CKD. Pharmacological strategies to decrease sEH activity might decrease postoperative AKI.</p></div