Both single and multicellular organisms depend on anti-stress mechanisms that enable them to deal with sudden changes in the environment, including exposure to heat and oxidants. Central to the ...stress response are dynamic changes in metabolism, such as the transition from the glycolysis to the pentose phosphate pathway-a conserved first-line response to oxidative insults
. Here we report a second metabolic adaptation that protects microbial cells in stress situations. The role of the yeast polyamine transporter Tpo1p
in maintaining oxidant resistance is unknown
. However, a proteomic time-course experiment suggests a link to lysine metabolism. We reveal a connection between polyamine and lysine metabolism during stress situations, in the form of a promiscuous enzymatic reaction in which the first enzyme of the polyamine pathway, Spe1p, decarboxylates lysine and forms an alternative polyamine, cadaverine. The reaction proceeds in the presence of extracellular lysine, which is taken up by cells to reach concentrations up to one hundred times higher than those required for growth. Such extensive harvest is not observed for the other amino acids, is dependent on the polyamine pathway and triggers a reprogramming of redox metabolism. As a result, NADPH-which would otherwise be required for lysine biosynthesis-is channelled into glutathione metabolism, leading to a large increase in glutathione concentrations, lower levels of reactive oxygen species and increased oxidant tolerance. Our results show that nutrient uptake occurs not only to enable cell growth, but when the nutrient availability is favourable it also enables cells to reconfigure their metabolism to preventatively mount stress protection.
Background
Cardiovascular disease (CVD) is the worldwide leading cause of morbidity and mortality. An early risk detection of apparently healthy people before CVD onset has clinical relevance in the ...prevention of cardiovascular events. We evaluated the association between the product of fasting plasma glucose and triglycerides (TyG index) and CVD.
Material and methods
A total of 5014 patients of the Vascular Metabolic CUN cohort (VMCUN cohort) were followed up during a median period of 10 years. We used a Cox proportional‐hazard ratio with repeated measures to estimate the risk of incidence of CVD across quintiles of the TyG index, calculated as lnfasting triglycerides (mg/dL) × fasting plasma glucose (mg(dL)/2, and plotted a receiver‐operating characteristics (ROC) curve to compare a prediction model fitted on the variables used in the Framingham risk score, a new model containing the Framingham variables with the TyG index, and the risk of coronary heart disease.
Results
A higher level of TyG index was significantly associated with an increased risk of developing CVD independent of confounding factors with a value of 2·32 (95% CI: 1·65–3·26) for those in the highest quintile and 1·52 (95% CI: 1·07–2·16) for those in the fourth quintile. The areas under the curve (AUC) of the ROC plots were 0·708 (0·68–0·73) for the Framingham model and 0·719 (0·70–0·74) for the Framingham + TyG index model (P = 0·014).
Conclusions
The TyG index, a simple measure reflecting insulin resistance, might be useful to early identify individuals at a high risk of developing a cardiovascular event.
There is limited information on the performance of rapid antigen detection (RAD) tests to identify SARS-CoV-2-infected asymptomatic individuals. In this field study, we evaluated the Panbio™ COVID-19 ...Ag Rapid Test Device (Abbott Diagnostics, Jena, Germany) for this purpose.
A total of 634 individuals (355 female; median age, 37 years; range, 9–87) were enrolled. Two nasopharyngeal swabs were collected from household (n = 338) and non-household contacts (n = 296) of COVID-19 cases. RAD testing was carried out at the point of care. The RT-PCR test used was the TaqPath COVID-19 Combo Kit (Thermo Fisher Scientific, MA, USA).
Household contacts were tested at a median of 2 days (range, 1–7) after diagnosis of the index case, whereas non-household contacts (n = 296) were tested at a median of 6 days (range, 1–7) after exposure. In total, 79 individuals (12.4%) tested positive by RT-PCR, of whom 38 (48.1%) yielded positive RAD results. The overall sensitivity and specificity of the RAD test was 48.1% (95% CI 37.4–58.9) and 100% (95% CI 99.3–100), respectively. Sensitivity was higher in household (50.8%; 95% CI 38.9–62.5) than in non-household (35.7%; 95% CI 16.3–61.2%) contacts. Individuals testing positive by RAD test were more likely (p < 0.001) to become symptomatic than their negative counterparts.
The Panbio test displays low sensitivity in asymptomatic close contacts of COVID-19 patients, particularly in non-household contacts. Nonetheless, establishing the optimal timing for upper respiratory tract collection in this group seems imperative to pinpoint test sensitivity.
To introduce methods for living guidelines based on practical experiences by the Australian Living Evidence Consortium (ALEC), the National Institute of Health and Care Excellence (NICE), and the ...Infectious Diseases Society of America (IDSA), with methodological support from the US Grading of Recommendations, Assessment, Development and Evaluations (GRADE) Network.
Members of ALEC, NICE, and the US GRADE Network, convened a working group to share experiences of the methods used to develop living guidelines and outline the key differences between traditional and living guidelines methods.
The guidance includes the following steps: 1) deciding if the guideline is a priority for a living approach, 2) preparing for living guideline development, 3) literature surveillance and frequency of searching, 4) assessment and synthesis of the evidence, 5) publication and dissemination, and 6) transitioning recommendations out of living mode.
This paper introduces methods for living guidelines and provides examples of the similarities and differences in approach across multiple organizations conducting living guidelines. It also introduces a series of papers exploring methods for living guidelines based on our practical experiences, including consumer involvement, selecting and prioritizing questions, search decisions, and methods decisions.
Display omitted
To describe the key features of a continual evidence surveillance process that can be implemented for living guidelines and to outline the considerations and trade-offs in adopting different ...approaches.
Members of the Australian Living Evidence Consortium (ALEC), National Institute of Health and Care Excellence (NICE), and the US GRADE Network (USGN) shared their practical experiences of and approaches to establishing surveillance systems for living guidelines. We identified several common components of evidence surveillance and listed the key features and considerations for each component drawn from case studies, highlighting differences with standard guidelines.
We developed guidance that covers the initial information needed to support decisions around suitability for living mode and the practical considerations in setting up continual search surveillance systems (search frequency, sources to search, use of automation, reporting the search, ongoing resources, and evaluation). The case studies draw on our experiences with developing guidelines for COVID-19, as well as for other conditions such as stroke and diabetes, and cover a range of practical approaches, including the use of automation.
This paper highlights different approaches to continual evidence surveillance that can be implemented in living guidelines.
Display omitted
SUMMARY
The stilbenoid pathway is responsible for the production of resveratrol in grapevine (Vitis vinifera L.). A few transcription factors (TFs) have been identified as regulators of this pathway ...but the extent of this control has not been deeply studied. Here we show how DNA affinity purification sequencing (DAP‐Seq) allows for the genome‐wide TF‐binding site interrogation in grape. We obtained 5190 and 4443 binding events assigned to 4041 and 3626 genes for MYB14 and MYB15, respectively (approximately 40% of peaks located within −10 kb of transcription start sites). DAP‐Seq of MYB14/MYB15 was combined with aggregate gene co‐expression networks (GCNs) built from more than 1400 transcriptomic datasets from leaves, fruits, and flowers to narrow down bound genes to a set of high confidence targets. The analysis of MYB14, MYB15, and MYB13, a third uncharacterized member of Subgroup 2 (S2), showed that in addition to the few previously known stilbene synthase (STS) targets, these regulators bind to 30 of 47 STS family genes. Moreover, all three MYBs bind to several PAL, C4H, and 4CL genes, in addition to shikimate pathway genes, the WRKY03 stilbenoid co‐regulator and resveratrol‐modifying gene candidates among which ROMT2‐3 were validated enzymatically. A high proportion of DAP‐Seq bound genes were induced in the activated transcriptomes of transient MYB15‐overexpressing grapevine leaves, validating our methodological approach for delimiting TF targets. Overall, Subgroup 2 R2R3‐MYBs appear to play a key role in binding and directly regulating several primary and secondary metabolic steps leading to an increased flux towards stilbenoid production. The integration of DAP‐Seq and reciprocal GCNs offers a rapid framework for gene function characterization using genome‐wide approaches in the context of non‐model plant species and stands up as a valid first approach for identifying gene regulatory networks of specialized metabolism.
Significance Statement
Subgroup 2 R2R3 MYBs have been shown to upregulate stilbene synthase expression in grapevine; however, their broader roles in the regulation of secondary metabolism remained unclear. Combining whole genome gene co‐expression networks, cistrome data for MYB13/14/15, and MYB15 overexpression we identified new regulatory targets across stilbenoid, early phenylpropanoid and shikimate pathways. We demonstrate the validity of this approach via the biochemical characterization of new ROMT targets and provide the networks and cistrome data to the community.
Abstract Aims We evaluated the potential role of the triglyceride–glucose index (TyG index) as a predictor of diabetes in a White European cohort, and compared it to fasting plasma glucose (FPG) and ...triglycerides. Methods 4820 patients of the Vascular-Metabolic CUN cohort (VMCUN cohort) were examined and followed up for 8.84 years (± 4.39). We performed a Cox proportional hazard ratio with repeated-measures analyses to assess the risk of developing type 2 diabetes across quartiles of FPG, triglycerides and the TyG index (lnfasting triglycerides (mg/dl) × fasting plasma glucose (mg/dl)/2), and plotted a receiver operating characteristics (ROC) curve for discrimination. Results There were 332 incident cases of type 2 diabetes involving 43,197.32 person-years of follow-up. We observed a progressively increased risk of diabetes in subjects with TyG index levels of 8.31 or more. Among those with normal fasting glucose at baseline, < 100 mg/dl, subjects with the TyG index in the fourth quartile were 6.87 times more likely to develop diabetes (95% CI, 2.76–16.85; P for trend < 0.001), as compared with the bottom quartile. The areas under the ROC curves (95% CI) were 0.75 (0.70–0.81) for TyG index, 0.66 (0.60–0.72) for FPG and 0.71 (0.65–0.77) for TG, in subjects with normal fasting glucose (p = 0.017). Conclusions Our data suggest that the TyG index is useful for the early identification of individuals at risk of type 2 diabetes. The TyG index seems to be a better predictor than FPG or triglycerides of the potential development of type 2 diabetes in normoglycemic patients.
Febrile seizure is one of the most common convulsive disorders in children. The neuromodulator adenosine exerts anticonvulsant actions through binding adenosine receptors. Here, the impact of ...hyperthermia‐induced seizures on adenosine A1 and A2A receptors and 5′‐nucleotidase activity has been studied at different periods in the cerebral cortical area by using radioligand binding, real‐time PCR, and 5′‐nucleotidase activity assays. Hyperthermic seizures were induced in 13‐day‐old rats using a warmed air stream from a hair dryer. Neonates exhibited rearing and falling over associated with hindlimb clonus seizures (stage 5 on Racine scale criteria) after hyperthermic induction. A significant increase in A1 receptor density was observed using 3HDPCPX as radioligand, and mRNA coding A1 was observed 48 h after hyperthermia‐induced seizures. In contrast, a significant decrease in A2A receptor density was detected, using 3HZM241385 as radioligand, 48 h after hyperthermia‐evoked convulsions. These short‐term changes in A1 and A2A receptors were also accompanied by a loss of 5′‐nucleotidase activity. No significant variations either in A1 or A2A receptor density or 5′‐nucleotidase were observed 5 and 20 days after hyperthermic seizures. Taken together, both regulation of A1 and A2A receptors and loss of 5′‐nucleotidase in the cerebral cortex suggest the existence of a neuroprotective mechanism against seizures.
Febrile seizure is one of the most common convulsive disorders in children. The consequences of hyperthermia‐induced seizures (animal model of febrile seizures) on adenosine A1 and A2A receptors and 5'‐nucleotidase activity have been studied at different periods in cerebral cortical area. A significant increase in A1 receptor density and mRNA coding A1 was observed 48 h after hyperthermia‐induced seizures. In contrast, a significant decrease in A2A receptor density and 5'‐nucleotidase activity was detected 48 h after convulsions evoked by hyperthermia. These changes suggest the possible existence of a neuroprotective mechanism against seizures.
Febrile seizure is one of the most common convulsive disorders in children. The consequences of hyperthermia‐induced seizures (animal model of febrile seizures) on adenosine A1 and A2A receptors and 5'‐nucleotidase activity have been studied at different periods in cerebral cortical area. A significant increase in A1 receptor density and mRNA coding A1 was observed 48 h after hyperthermia‐induced seizures. In contrast, a significant decrease in A2A receptor density and 5'‐nucleotidase activity was detected 48 h after convulsions evoked by hyperthermia. These changes suggest the possible existence of a neuroprotective mechanism against seizures.
Invasive fungal disease accounts for about 3.8 million deaths annually, an unacceptable rate that urgently prompts the discovery of new knowledge‐driven treatments. We report the use of camelid ...single‐domain nanobodies (Nbs) against fungal β‐1,3‐glucanosyltransferases (Gel) involved in β‐1,3‐glucan transglycosylation. Crystal structures of two Nbs with Gel4 from Aspergillus fumigatus revealed binding to a dissimilar CBM43 domain and a highly conserved catalytic domain across fungal species, respectively. Anti‐Gel4 active site Nb3 showed significant antifungal efficacy in vitro and in vivo prophylactically and therapeutically against different A. fumigatus and Cryptococcus neoformans isolates, reducing the fungal burden and disease severity, thus significantly improving immunocompromised animal survival. Notably, C. deneoformans (serotype D) strains were more susceptible to Nb3 and genetic Gel deletion than C. neoformans (serotype A) strains, indicating a key role for β‐1,3‐glucan remodelling in C. deneoformans survival. These findings add new insight about the role of β‐1,3‐glucan in fungal biology and demonstrate the potential of nanobodies in targeting fungal enzymes to combat invasive fungal diseases.
Nanobodies show promise in treating invasive fungal diseases such as aspergillosis and cryptococcosis. By selectively targeting enzymes critical for fungal cell wall remodeling, these single‐domain antibodies significantly enhanced survival rates in animal models. This approach opens up a promising new pathway for the development of antifungal therapies, potentially improving outcomes for immunocompromised patients.
This article is part of a series on methods for living guidelines, consolidating practical experiences from developing living guidelines. It focuses on methods for identification, selection, and ...prioritization of clinical questions for a living approach to guideline development.
Members of the Australian Living Evidence Consortium, the National Institute of Health and Care Excellence and the US Grading of Recommendations, Assessment, Development and Evaluations Network, convened a working group. All members have expertize and practical experience in the development of living guidelines. We collated methods, documents on prioritization from each organization's living guidelines, conducted interviews and held working group discussions. We consolidated these to form best practice principles which were then edited and agreed on by the working group members.
We developed best practice principles for (1) identification, (2) selection, and (3) prioritization, of questions for a living approach to guideline development. Several different strategies for undertaking prioritizing questions are explored.
The article provides guidance for prioritizing questions in living guidelines. Subsequent articles in this series explore consumer involvement, search decisions, and methods decisions that are appropriate for questions with different priority levels.
PDF
