Structural and kinetic characterization of DUSP5 with a Di-phosphorylated tripeptide substrate from the ERK activation loop.

Introduction: Dual specific phosphatases (DUSPs) are mitogen-activated protein kinase (MAPK) regulators, which also serve as drug targets for treating various vascular diseases. Previously, we have presented mechanistic characterizations of DUSP5 and its interaction with pERK, proposing a dual active site.

Methods: Herein, we characterize the interactions between the DUSP5 phosphatase domain and the pT-E-pY activation loop of ERK2, with specific active site assignments.

Application of the revised criteria for diagnosis and staging of Alzheimer's disease: Drug development and clinical practice.

Unlabelled: The newly proposed revised criteria for diagnosis and staging of Alzheimer's disease (AD) by the Alzheimer's Association (AA) Workgroup represent a significant milestone in the field. These criteria offer objective measures for diagnosing and staging biological AD, bridging the gap between research and clinical care. Although implementation feasibility may vary across regions and settings, improving the availability and accuracy of biomarkers, especially plasma biomarkers, is expected to enhance the applicability of these criteria in clinical practice.

Recombinant Fungal Aspartic Endopeptidases: Insights into Protein Hydrolysis and Combined Effect with Pepsin for Animal Feed Application.

Protein hydrolysis under acidic conditions can improve the product quality, nutrient availability, and cost efficiency, particularly when neutral or alkaline enzymes are ineffective. Six fungal aspartic endopeptidases (FAPs) were recombinantly expressed as active enzymes in , with peak activity between 30-50 °C and pH 3.0-4.

Rational engineering of minimally immunogenic nucleases for gene therapy.

Genome editing using CRISPR-Cas systems is a promising avenue for the treatment of genetic diseases. However, cellular and humoral immunogenicity of genome editing tools, which originate from bacteria, complicates their clinical use. Here we report reduced immunogenicity (Red)(i)-variants of two clinically relevant nucleases, SaCas9 and AsCas12a.

The spatially informed mFISHseq assay resolves biomarker discordance and predicts treatment response in breast cancer.

Current assays fail to address breast cancer's complex biology and accurately predict treatment response. On a retrospective cohort of 1082 female breast tissues, we develop and validate mFISHseq, which integrates multiplexed RNA fluorescent in situ hybridization with RNA-sequencing, guided by laser capture microdissection. This technique ensures tumor purity, unbiased whole transcriptome profiling, and explicitly quantifies intratumoral heterogeneity.

AI-based visualization of loose connective tissue as a dissectable layer in gastrointestinal surgery.

We aimed to develop an AI model that recognizes and displays loose connective tissue as a dissectable layer in real-time during gastrointestinal surgery and to evaluate its performance, including feasibility for clinical application. Training data were created under the supervision of gastrointestinal surgeons. Test images and videos were randomly sampled and model performance was evaluated visually by 10 external gastrointestinal surgeons.

Lossless and reference-free compression of FASTQ/A files using GeneSqueeze.

As sequencing becomes more accessible, there is an acute need for novel compression methods to efficiently store sequencing files. Omics analytics can leverage sequencing technologies to enhance biomedical research and individualize patient care, but sequencing files demand immense storage capabilities, particularly when sequencing is utilized for longitudinal studies. Addressing the storage challenges posed by these technologies is crucial for omics analytics to achieve their full potential.

Correlation of triglyceride glucose index with all cause mortality in acute myocardial infarction patients following percutaneous coronary intervention.

The Triglyceride glucose (TyG) index is a dependable indicator of IR, with numerous studies underscoring its influence on Cardiovascular disease. Nevertheless, the connection between the TyG index and prognosis in AMI patients after PCI is still uncertain. This investigation aims to explore the link in individuals who have received PCI for AMI.

Uricase from , a Candidate for Industrial Application of Reducing Uric Acid Content of Bean Products.

Microbial uricase is an essential enzyme in purine degradation and the development of low-purine food. High enzyme activity and an appropriate optimum pH must be established for low-purine food. Uricases from , , , , and were heterologously expressed in .

Understanding Noninferiority Trials: What Stroke Specialists Should Know.

Noninferiority trials aim to prove that the efficacy, defined in terms of a key clinical outcome, of a new treatment is not meaningfully worse than that of an established active control. Noninferiority trials are important when other aspects of care can be improved, such as convenience, toxicity, costs, and safety (nonefficacy benefits). While the motivation for a noninferiority trial is straightforward, the design, execution, and interpretation of these trials is not a trivial task.

[Molecular Mechanism of Protein C Deficiency Caused by Mutations of Gene N355S, G392E, T314A].

Objective: To study the molecular mechanism of functional defect of protein C (PC) caused by point mutations of human protein C gene ( ) N355S , G392E and T314A.

Methods: The wild-type and mutant plasmids (PC, PC, PC, PC) of gene were constructed and transiently transfected into HEK293 cells. The expression of mutant proteins in vitro were tested.

Galactokinase and galactose metabolism in Leishmania spp.

In Leishmania, the nucleotide-sugar UDP-galactose can be synthesized by a salvage pathway, the Isselbacher route, involving phosphorylation of galactose and the action of UDP-sugar pyrophosphorylase. The first enzyme of the pathway, galactokinase, has yet to be studied in this parasite. Here, we report a molecular and biochemical characterization of this enzyme in Leishmania mexicana.

Biochemical characterization and inhibitor potential of African swine fever virus thymidine kinase.

African Swine Fever (ASF) is a highly contagious disease affecting both domestic pigs and wild boars. In domestic pigs, ASF is a rapidly-progressing disease with a mortality rate reaching 100 %, causing tremendous economic loss in affected areas. ASFV is caused by African Swine Fever Virus (ASFV), which is a large, enveloped double-stranded DNA virus belonging to the Asfarviridae family.

Semi-rational engineering of 7β-hydroxysteroid dehydrogenase enhances forward reaction activity towards ursodeoxycholic acid synthesis.

7β-Hydroxysteroid dehydrogenase (7β-HSDH) catalyzes the reversible reaction between 7-ketolithocholic acid (7K-LCA) and ursodeoxycholic acid (UDCA). However, its much lower forward reaction activity led to the unsatisfactory UDCA production. Here, by autodocking 7K-LCA and UDCA into the structure of Hyphomicrobium sp.

Ancestral Nitrilase Mining and Semi-Rational Engineering for Enhanced Thermal Stability in Rapeseed Meals-Derived Nitriles Degradation.

Rapeseed meal (RSM), a protein-rich byproduct, holds potential as a high-quality animal feed, but nitrile compounds derived from glucosinolates (GSLs) in RSM pose a toxicity risk. Nitrilases, enzymes that hydrolyze toxic nitriles to carboxylic acids, offer a potential solution for detoxification. However, the low thermal stability of nitrilases restricts their industrial applicability.

Use of Internet/Application-Based E-consults by General Practitioners in Japan to Resolve Patients' Problems: A Descriptive Observational Study.

Introduction Electronic consultations (e-consults) refer to digital health technologies that enhance clinical information sharing and improve patients' access to expert healthcare providers. This study aims to describe the current utilization of the Internet/application-based e-consult system and to assess how effectively it resolves clinical questions. Methods Participants were general practitioners (GPs) who had installed the e-consult platform on their personal computers or cell phones.

An isothermal calorimetry assay for determining steady state kinetic and Ensitrelvir inhibition parameters for SARS-CoV-2 3CL-protease.

This manuscript details the application of Isothermal Titration Calorimetry (ITC) to characterize the kinetics of 3CL, the main protease from the Severe Acute Respiratory Syndrome CoronaVirus-2 (SARS-CoV-2), and its inhibition by Ensitrelvir, a known non-covalent inhibitor. 3CL is essential for producing the proteins necessary for viral infection, which led to the COVID-19 pandemic. The ITC-based assay provided rapid and reliable measurements of 3CL activity, allowing for the direct derivation of the kinetic enzymatic constants K and k by monitoring the thermal power required to maintain a constant temperature as the substrate is consumed.

A multiplex microarray lateral flow immunoassay device for simultaneous determination of five mycotoxins in rice.

Co-occurrence of multiple mycotoxins is a growing global food safety concern due to their harmful effects on humans and animals. This study developed an eco-friendly sample preparation method and an innovative multiplex microarray-based lateral flow immunoassay, using a novel portable reader for on-site simultaneous determination of five regulated mycotoxins-aflatoxin B, T-2 toxin, zearalenone, deoxynivalenol, and fumonisin B in rice. The eco-friendly and ultrafast extraction procedure utilizes a bio-based solvent.

Correction: Absence of posterior pituitary bright spot in adults with CNS tuberculosis: A case-control study.

[This corrects the article DOI: 10.1371/journal.pone.

Urinary albumin-to-creatinine ratio for predicting risk of all-cause mortality and specific-cause mortality in patients with rheumatoid arthritis: evidence from NHANES 1999-2018.

Objective: To explore the relationship between urinary albumin-to-creatinine ratio (uACR) and all-cause/specific-cause mortality among patients with rheumatoid arthritis (RA).

Methods: This study included 1354 RA patients in the National Health and Nutritional Examination Surveys (NHANESs) during 1999-2018. The mortality status was assessed by linkage to death certificate data reported in the National Death Index (NDI) until December 31, 2019.

The Cryo-EM structure of human CD163 bound to haptoglobin-hemoglobin reveals molecular mechanisms of hemoglobin scavenging.

CD163, a macrophage-specific receptor, plays a critical role in scavenging hemoglobin released during hemolysis, protecting against oxidative effects of heme iron. In the bloodstream, hemoglobin is bound by haptoglobin, leading to its immediate endocytosis by CD163. While haptoglobin's structure and function are well understood, CD163's structure and its interaction with the haptoglobin-hemoglobin complex have remained elusive.