Assembly and architecture of endogenous NMDA receptors in adult cerebral cortex and hippocampus.

The cerebral cortex and hippocampus are crucial brain regions for learning and memory, which depend on activity-induced synaptic plasticity involving N-methyl-ᴅ-aspartate receptors (NMDARs). However, subunit assembly and molecular architecture of endogenous NMDARs (eNMDARs) in the brain remain elusive. Using conformation- and subunit-dependent antibodies, we purified eNMDARs from adult rat cerebral cortex and hippocampus.

Development of Synthetic Antimicrobial Peptides Based on Genomic Analysis of Streptococcus salivarius.

Background: In the oral environment, the production of bacteriocins or antimicrobial peptides (AMPs) plays a crucial role in maintaining ecological balance by impeding the proliferation of closely related microorganisms. This study aims to conduct in silico genome screening of Streptococcus salivarius to identify potential antimicrobial compounds existing as hypothetical peptides, with the goal of developing novel synthetic antimicrobial peptides.

Methods: Draft genomes of various oral Streptococcus salivarius strains were obtained from the NCBI database and subjected to analysis using bioinformatic tools, viz.

Deep-Learning-Assisted Digital Fluorescence Immunoassay on Magnetic Beads for Ultrasensitive Determination of Protein Biomarkers.

Digital fluorescence immunoassay (DFI) based on random dispersion magnetic beads (MBs) is one of the powerful methods for ultrasensitive determination of protein biomarkers. However, in the DFI, improving the limit of detection (LOD) is challenging since the ratio of signal-to-background and the speed of manual counting beads are low. Herein, we developed a deep-learning network (ATTBeadNet) by utilizing a new hybrid attention mechanism within a UNet3+ framework for accurately and fast counting the MBs and proposed a DFI using CdS quantum dots (QDs) with narrow peak and optical stability as reported at first time.

Application of Principal Component Analysis as a Prediction Model for Feline Sporotrichosis.

Sporotrichosis is a worldwide zoonotic disease that is spreading and causing epidemics in large urban centers. Cats are the most susceptible species to develop the disease, which could cause significant systemic lesions. The aim was to investigate and to identify predictive indicators of disease progression by correlations between the blood profile (hematological and biochemical analytes) and cutaneous lesion patterns of 70 cats diagnosed with .

Protective Efficacy of Decreasing Antigen Doses of a Subcellular Vaccine Against Ovine Enzootic Abortion in a Pregnant Sheep Challenge Model.

Background/objective: , the cause of ovine enzootic abortion, is a zoonotic bacterial pathogen and one of the most infectious causes of foetal death in sheep worldwide. Although the disease can be controlled using commercial inactivated and live whole-organism vaccines, there are issues with both, particularly concerning efficacy and safety. Recently, we have described the development of a new COMC (chlamydial outer membrane complex) vaccine based on a detergent-extracted outer membrane protein preparation of the pathogen, which can be delivered in a single inoculation and is both efficacious and safe.

'Ship-in-a-Bottle' Integration of pH-Sensitive 3D Proteinaceous Meshes into Microfluidic Channels.

Microfluidic sensors incorporated onto chips allow sensor miniaturization and high-throughput analyses for point-of-care or non-clinical analytical tools. Three-dimensional (3D) printing based on femtosecond laser direct writing (fs-LDW) is useful for creating 3D microstructures with high spatial resolution because the structures are printed in 3D space along a designated laser light path. High-performance biochips can be fabricated using the 'ship-in-a-bottle' integration technique, in which functional microcomponents or biomimetic structures are embedded inside closed microchannels using fs-LDW.

Enhanced Electrochemiluminescence from Ruthenium-Tagged Immune Complex at Flexible Chains for Sensitive Analysis of Glutamate Decarboxylase Antibody.

Herein, a sensitive electrochemiluminescence (ECL) immunosensor is designed by immobilizing ruthenium-tagged immune complexes at flexible poly-ethylene-glycol (PEG) chains on the electrode surface, which offers more freedom for the collision of the ruthenium complex at the electrode during the initial ECL reaction. The electrochemical characterizations confirm the loose structure of the assembled layer with the immune complex, providing an increase in the current and the resultant enhanced ECL emissions. Comparing the sensors with the rigid structure, a 34-fold increase in the maximal ECL emission is recorded when PEG3400 is used as a linker.

Carboxylated Graphene: An Innovative Approach to Enhanced IgA-SARS-CoV-2 Electrochemical Biosensing.

Biosensors harness biological materials as receptors linked to transducers, enabling the capture and transformation of primary biorecognition signals into measurable outputs. This study presents a novel carboxylation method for synthesizing carboxylated graphene (CG) under acidic conditions, enhancing biosensing capabilities. The characterization of the CG was performed using scanning electron microscopy (SEM), energy-dispersive X-ray spectroscopy (EDS), Raman spectroscopy, thermogravimetric analysis (TGA), and X-ray diffraction (XRD).

Fully Inkjet-Printed Flexible Graphene-Prussian Blue Platform for Electrochemical Biosensing.

Prussian Blue (PB) is commonly incorporated into screen-printed enzymatic devices since it enables the determination of the enzymatically produced hydrogen peroxide at low potentials. Inkjet printing is gaining popularity in the development of electrochemical sensors as a substitute for screen printing. This work presents a fully inkjet-printed graphene-Prussian Blue platform, which can be paired with oxidase enzymes to prepare a biosensor of choice.

A Truncated Multi-Thiol Aptamer-Based SARS-CoV-2 Electrochemical Biosensor: Towards Variant-Specific Point-of-Care Detection with Optimized Fabrication.

With the goal of fast and accurate diagnosis of infectious diseases, this study presents a novel electrochemical biosensor that employs a refined aptamer (C9t) for the detection of spike (S) protein SARS-CoV-2 variants in a flexible multielectrode aptasensor array with PoC capabilities. Two aptamer modifications were employed: removing the primer binding sites and including two dithiol phosphoramidite anchor molecules. Thus, reducing fabrication time from 24 to 3 h and increasing the stability and sparseness for multi-thiol aptasensors compared to a standard aptasensor using single thiols, without a reduction in aptamer density.

Microspectrometer-Enabled Real-Time Concentration Monitoring in the Microfluidic Protein Enrichment Chip.

This study presents a novel microspectrometer-integrated microfluidic system for real-time protein concentration monitoring. The device employs electrokinetic principles for efficient protein preconcentration in a PDMS and Nafion film channel. Using FITC-labeled BSA as a model protein, the system demonstrated a linear correlation between protein concentration and absorbance at 491 nm.

Integrated SegFlow, µSIA, and UPLC for Online Sialic Acid Quantitation of Glycoproteins Directly from Bioreactors.

This study emphasizes the critical importance of closely monitoring and controlling the sialic acid content in therapeutic glycoproteins, including EPO, interferon-γ, Orencia, Enbrel, and others, as the level of sialylation directly impacts their pharmacokinetics (PK), immunogenicity, potency, and overall clinical performance due to its influence on protein clearance via hepatic asialoglycoprotein receptors (ASGPR). The ASGPR recognizes and binds to glycoproteins exposed to terminal galactose or N-acetylgalactosamine residues, leading to receptor-mediated endocytosis. Recent studies have demonstrated that sialylation of O-linked glycan plays a role in protecting against macrophage galactose lectin (MGL)-mediated clearance.

Elevated A2F bisect N-glycans of serum IgA reflect progression of liver fibrosis in patients with MASLD.

Background: Advanced liver fibrosis in cases of metabolic dysfunction-associated steatotic liver disease (MASLD) leads to cirrhosis and hepatocellular carcinoma. The current gold standard for liver fibrosis is invasive liver biopsy. Therefore, a less invasive biomarker that accurately reflects the stage of liver fibrosis is highly desirable.

YAT2150 is irresistible in Plasmodium falciparum and active against Plasmodium vivax and Leishmania clinical isolates.

We recently characterized the potent antiplasmodial activity of the aggregated protein dye YAT2150, whose presumed mode of action is the inhibition of protein aggregation in the malaria parasite. Using single-dose and ramping methods, assays were done to select Plasmodium falciparum parasites resistant to YAT2150 concentrations ranging from 3× to 0.25× the in vitro IC of the compound (in the two-digit nM range) and performed a cross-resistance assessment in P.

LC-MS/MS based analytical strategies for the detection of lipid peroxidation products in biological matrices.

Oxidative stress (OS) arises mainly from exposure to reactive oxygen species (ROS) such as superoxide anion, hydroxyl radical, and hydrogen peroxide. These molecules can cause significant damage to proteins, DNA, and lipids, leading to various diseases. Cells fight ROS with detoxifying enzymes; however, an imbalance can cause damage leading to ischemic conditions, heart disease progression, and neurological disorders such as Alzheimer's disease.

Genetic susceptibility and potential therapeutic targets of unruptured intracranial aneurysms: A genome-wide study based on Mendelian randomization.

Background: At present, although some studies have offered certain insights into the genetic factors related to unruptured intracranial aneurysms (uIAs), the potential genetic targets associated with uIAs remain largely unknown. Thus, this research adopted Mendelian randomization (MR) analysis to study two genome-wide association studies on uIAs, aiming to determine the reliable genetic susceptibility and potential therapeutic targets for uIAs.

Methods: This study summarizes the data of expression quantitative trait loci (eQTL) as exposure data.

Capillary Electrophoresis-Mass Spectrometry for Top-Down Proteomics.

Mass spectrometry (MS)-based top-down proteomics (TDP) characterizes proteoforms in cells, tissues, and biological fluids (e.g., human plasma) to better our understanding of protein function and to discover new protein biomarkers for disease diagnosis and therapeutic development.

Microfluidic purification of genomic DNA.

We describe a microfluidic device to extract DNA from a cell lysate, without the need for centrifuges, magnetic beads, or gels. Instead, separation is driven by transverse migration of DNA, which occurs when a polyelectrolyte solution flowing through a microfluidic channel is subjected to an electric field. The coupling of the weak shearing with the axial electric field is highly selective for long, flexible, charged molecules, of which DNA is the sole example in a typical cell lysate.

Elucidating the wedelolactone biosynthesis pathway from Eclipta prostrata (L.) L.: a comprehensive analysis integrating de novo comparative transcriptomics, metabolomics, and molecular docking of targeted proteins.

Eclipta prostrata belongs to the Asteraceae family. The plant contains bioactive compounds like wedelolactone (WDL) and demethylwedelolactone (DW). Its transcriptomic information engaged with secondary metabolite biosynthesis is not available.

Highly Sensitive Digital and Analog Immunoassay Based on Submicron Magnetic Beads and Fluorescent Microspheres.

The detection of disease-related protein biomarkers plays a crucial role in the early diagnosis, treatment, and monitoring of diseases. The concentrations of protein biomarkers can vary significantly in different diseases or stages of the same disease. However, most of the existing analytical methods cannot simultaneously meet the requirements of high sensitivity and a wide dynamic range.

Dual surrogate imprinting: an innovative strategy for the preparation of virus-selective particles.

This work involves the preparation of dual surrogate-imprinted polymers (D-MIPs) for the capture of SARS-CoV-2. To achieve this goal, an innovative and novel dual imprinting approach using carboxylated-polystyrene (PS-COOH) nanoparticles with a diameter of 100 nm and a SARS-CoV-2 Spike-derived peptide was carried out at the surface of amine-functionalized silica (PS-NH) microspheres with a diameter of 500 nm. Firstly, PS-COOH nanoparticles with the same size and spherical shape as the SARS-CoV-2 virus were employed to form hemispherical indentations (HI) at the surface of the PS-NH microspheres (obtaining dummy particle-imprinted polymers, HI-MIPs).

Advancing Newborn Screening in Washington State: A Novel Multiplexed LC-MS/MS Proteomic Assay for Wilson Disease and Inborn Errors of Immunity.

For many genetic disorders, there are no specific metabolic biomarkers nor analytical methods suitable for newborn population screening, even where highly effective preemptive treatments are available. The direct measurement of signature peptides as a surrogate marker for the protein in dried blood spots (DBSs) has been shown to successfully identify patients with Wilson Disease (WD) and three life-threatening inborn errors of immunity, X-linked agammaglobulinemia (XLA), Wiskott-Aldrich syndrome (WAS), and adenosine deaminase deficiency (ADAD). A novel proteomic-based multiplex assay to detect these four conditions from DBS using high-throughput LC-MS/MS was developed and validated.

Antidiabetic Effects of Quercetin and Silk Sericin in Attenuating Dysregulation of Hepatic Gluconeogenesis in Diabetic Rats Through Potential Modulation of PI3K/Akt/FOXO1 Signaling: In Vivo and In Silico Studies.

Type 2 diabetes mellitus (T2DM) is an intricate disease correlated with many metabolic deregulations, including disordered glucose metabolism, oxidative stress, inflammation, and cellular apoptosis due to hepatic gluconeogenesis aberrations. However, there is no radical therapy to inhibit hepatic gluconeogenesis disturbances yet. We thus sought to probe the effectiveness and uncover the potential mechanism of quercetin (QCT) and silk sericin (SS) in mitigating hyperglycemia-induced hepatic gluconeogenesis disorder, which remains obscure.

Identify biological Alzheimer's disease using a novel nucleic acid-linked protein immunoassay.

Blood-based biomarkers have been revolutionizing the detection, diagnosis and screening of Alzheimer's disease. Specifically, phosphorylated-tau variants (p-tau, p-tau and p-tau) are promising biomarkers for identifying Alzheimer's disease pathology. Antibody-based assays such as single molecule arrays immunoassays are powerful tools to investigate pathological changes indicated by blood-based biomarkers and have been studied extensively in the Alzheimer's disease research field.

Maturing conditions of bimetallic nanocomposites as a new factor influencing Au-Ag synergism and impact of Cu(II) and/or Fe(III) on luminescence.

Gold-silver synergism has been well documented in many scientific works dealing with luminescent nanostructures that are exploitable in biomedical and environmental application. Frequently, the ratio of Au : Ag in synthetic mixtures was varied to influence the extent of Au-Ag synergism of the resulting luminescent gold-silver nanoclusters (GSNCs). However, in our approach, a new step, maturing under differing conditions using the same Au : Ag ratio (5 : 1), has been investigated systematically for the very first time.