Browse Articles

A targeted tiled amplicon sequencing approach for clade and subclade level differentiation of monkeypox virus from wastewater

Abstract Wastewater-based surveillance (WBS) has modernized in recent years and emerged as an important tool for the monitoring of viral pathogens, including monkeypox virus (MPXV). Here we describe a novel targeted amplicon sequencing method developed for clade and subclade characterization of MPXV from municipal wastewater. This new method addresses the limitations of PCR-based methods and the challenges of sequencing a pathogen displaying low viral load in municipal wastewater samples. A tiled amplicon scheme composed of 11 primer pairs targeting a 4.2 kb portion of the inverted terminal repeat (ITR) region of the MPXV genome was designed and tested. In silico analysis demonstrated high accuracy for clade and subclade calls using the full target region, with specific amplicons also exhibiting strong performance individually. An MPXV consensus sequence representing the entire target region was successfully sequenced from a wastewater sample and differentiated from positive controls by a distinct deletion within a short homopolymeric region. Notably, clade-informing data was also achieved from partial sequences recovered from lower abundance samples. This study presents a new sequencing method targeting MPXV with enhanced genomic resolution compared to existing PCR-based approaches, providing critical genomic-level information informing MPXV surveillance and public health interventions.

MANIT: a multilayer ANN integrated framework using biometrics and historical features for online examination proctoring

Progressive multi-scale multi-attention fusion for hyperspectral image classification

Transfer accuracy and chairside efficiency of rigid- and flexible-printed versus double vacuum-formed lingual indirect bonding trays: a prospective cohort study

Abstract This study aimed to compare bracket-transfer accuracy, chairside time, and early bond failure among rigid 3D-printed (RP), flexible 3D-printed (FP), and double vacuum-formed (DV) lingual indirect bonding trays. Thirty-three consecutive adults (n = 11) were prospectively enrolled. After virtual setup and tray fabrication, brackets were bonded following a standard protocol. Post-bonding intraoral scans were superimposed on the planned setup, and six positional discrepancies (mesial-distal, in-out, height, rotation, tip, and torque) were quantified for each tooth. One-sample Wilcoxon signed-rank tests compared observed errors with clinical limits (0.5 mm, 2.0°). Kruskal-Wallis and pairwise Mann-Whitney U tests assessed inter-tray differences. 818 brackets (RP = 280; FP = 259; DV = 279) were analysed. Median translational errors were ≤ 0.10 mm for RP/FP and ≤ 0.07 mm for DV; all were below the 0.5 mm threshold (P < 0.001). Median pure-rotation errors remained ≤ 1.0° for all trays. Tip met the 2.0° limit except for FP in the lower-anterior segment (P = 0.184). Torque rarely met the limit, with RP-upper-anterior being the sole exception. DV outperformed RP and FP in mesial-distal accuracy across upper and lower-posterior regions (P ≤ 0.002). All tray systems delivered clinically acceptable translational accuracy, but DV trays provided the most precise mesial-distal positioning. Selecting a tray architecture that balances rigidity and elasticity can markedly improve chairside efficiency without compromising bracket-placement accuracy.

Meta-analysis of the effects of plyometric training on athletic performance in handball athletes

Discovery of potent endolysins against Pseudomonas aeruginosa and other gram-negative ESKAPE pathogens by an agar-based screening method

Construction of multiphysics coupling equations and study on flow field migration patterns in shallow buried goaf under air leakage conditions

Inspection of railway catenary systems using machine learning with domain knowledge integration

Bond behaviour between H-shaped steel and concrete after freeze-thaw cycles

Transcriptomic profiling of skeletal muscle in the DMDmdx rat model of Duchenne muscular dystrophy

Abstract Duchenne muscular dystrophy (DMD) is a severe X-linked recessive disorder caused by a mutation in the Dmd gene, leading to progressive muscle degradation, increasing weakness, and typically resulting in death before the third decade of life. To investigate the pathobiology of DMD, this study employed the Sprague-Dawley Dmd-mutated rat model (DMD mdx ) and analyzed gene expression profiles and pathological molecular pathways. The methods used included histopathological, biochemical, and transcriptomic analyses of dystrophic skeletal muscle from DMD mdx and wild-type (WT) individuals. Histological analysis of skeletal muscle tissue from DMD mdx rats revealed multifocal necrosis, fibrosis, and inflammation, whereas WT rats displayed normal muscle architecture. Biochemical analysis revealed significant alterations in plasma markers of muscle damage and metabolism in DMD mdx rats compared to WT controls, including elevated AST, ALT, ALP, CPK, and LDH levels. Additionally, oxidative status measurements showed reduced antioxidant capacity and increased lipid peroxidation in dystrophic skeletal muscle, as evidenced by lower TAS, GR, GPx, and SOD activities and higher TBARS levels. RNA-seq analysis identified 3,615 differentially expressed genes between the two groups, associated with muscle contraction, extracellular matrix (ECM) organization, and cytoskeleton organization. Notably, Dmd, Actc1, Col6a1, and Mmp2 were significantly downregulated. Gene ontology and pathway enrichment analyses indicated dystrophic changes in skeletal muscle, disruptions in calcium homeostasis, and alterations in actin cytoskeleton regulation. KEGG and Reactome pathway analyses revealed upregulation of the MAPK signaling and immune system pathways and downregulation of the ECM organization pathway. These findings support the hypothesis that targeting complex intracellular signaling pathways in DMD may represent a promising therapeutic strategy. Given that the DMD mdx rat model closely mimics human DMD pathology compared to other animal models, it offers a more realistic platform for studying the molecular mechanisms of the disease and improving the translational potential of therapeutic approaches.

Gradual transition of pyramidal cell types in the dorsal hippocampal area CA2b of the C57BL/6 mouse

Abstract Hippocampal area CA2 is important for social recognition memory. It has classically been defined as the region between areas CA3 and CA1 where pyramidal cells have larger cell bodies than CA1 neurons, but lack mossy fiber input and thorny excrescences (TEs) typical for CA3 neurons. Based on molecular signatures, the borders of area CA2 have been redefined, with area CA2b now covering parts of former area CA3a. Functional data suggest that CA2b is a mixture of CA3 and CA2 cells, yet information about the spatial distribution of these cells within CA2b remains vague. In the present study, we filled pyramidal cells in dorsal CA3-CA2-CA1, post-hoc identified CA2 cells using Purkinje Cell Protein 4 (PCP4), and analyzed their proximal apical spines using confocal and electron microscopy. We found that dorsal CA2b resembles an intermediate zone between CA3 and CA2a, where CA3 neurons with large TEs are gradually replaced by CA2 neurons. These CA2 neurons are heterogeneous in their scarce display of spines and larger spiny protrusions, rarely also carry TEs, and form synapses with mossy fibers. Thus, morphology needs to be combined with molecular markers to identify CA2 neurons in area CA2b with certainty.

Fractional-order model of malaria incorporating treatment and prevention strategies

Synthesis of carbonyl-functionalized mercaptosilsesquioxane

Abstract Commercially available and inexpensive potassium carbonate (K2CO3) has been applied to the hydrothiolation of mercaptopropylisobutyl POSS (SQ-SH) with α,β-unsaturated carbonyl compounds of different types. This innovative approach has been proved to be effective for a wide range of substrates, leading to novel classes of functionalized SQ-based materials in yields exceeding 90% under ambient temperature, air and transition metal-free conditions. Additionally, the proposed synthetic strategy has been used for the modification of chalcones, the compounds with significant medicinal and synthetic potential. As a result, twelve new mercapto-modified nanomaterials containing carbonyl groups were obtained and comprehensively characterized by spectroscopic methods and mass analysis. The selected product of great practical application was studied in terms of its thermal properties.

Exploration of autophagy-associated genes and potential molecular mechanisms in type 1 diabetes and osteoporosis

Multimodal anti fraud education improves cognitive emotional and behavioral engagement in older adults

Electrokinetic blood flow of Carreau ternary nanofluids in stenotic arteries with thermal reactions under CC heat flux for therapy

Evaluation of road performance and carbon emission accounting analysis of recycled aggregates from construction and demolition waste

Fermentation quality improvement of cigar wrapper inoculated with exogenous strain Staphylococcus capitis S1

Hysteresis in ocean export production owing to CO2 forcing

Induction of proline-rich proteins in response to tannin treatment in Caenorhabditis elegans

Abstract Salivary proline-rich proteins (PRPs) represent a common mechanism of defense against tannins in mammals. Few reports exist regarding the occurrence or PRPs with similar function in nematodes and none of these proteins or their coding genes have been functionally characterized so far. In Caenorhabditis elegans, two genes (clx-1 and T22D1.2) were strongly induced upon tannin treatment of the nematodes, both of them potentially encoding proline-rich proteins. Therefore, translation of these genes into proteins was confirmed and the expression pattern was investigated in more detail. Particularly T22D1.2 was found to be exclusively up-regulated in worms treated with test substances possessing astringent properties, especially tannins, whereas no expression was observed for any other stressor or in the untreated control group. Similar to mammalian PRPs, repetitive proline-rich sequences were identified in both of the corresponding proteins. A potential role in tannin defense was supported by an increased survival of tannin-treated worms when T22D1.2 was constitutively expressed under the vit-5 promoter. However, no differences were observed in the clx-1 and T22D1.2 knockout mutants in comparison to the wild type, respectively. Within the current study, evidence was provided for the existence of repetitive proline-rich proteins in the free-living nematode C. elegans, of which particularly T22D1.2 may be involved in tannin defense.