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Abstract The epitranscriptome embodies many new and largely unexplored functions of RNA. A significant roadblock hindering progress in epitranscriptomics is the identification of more than one modification in individual transcript molecules. We address this with CHEUI (CH3 (methylation) Estimation Using Ionic current). CHEUI predicts N6-methyladenosine (m6A) and 5-methylcytosine (m5C) in individual molecules from the same sample, the stoichiometry at transcript reference sites, and differential methylation between any two conditions. CHEUI processes observed and expected nanopore direct RNA sequencing signals to achieve high single-molecule, transcript-site, and stoichiometry accuracies in multiple tests using synthetic RNA standards and cell line data. CHEUI’s capability to identify two modification types in the same sample reveals a co-occurrence of m6A and m5C in individual mRNAs in cell line and tissue transcriptomes. CHEUI provides new avenues to discover and study the function of the epitranscriptome.

Objective: Prolonged immobilization often results in myogenic and arthrogenic contractures, with capsular fibrosis being the critical aspect of the latter. This study aimed to determine whether nicorandil can mitigate joint contracture and fibrosis by inhibiting the RhoA/ROCK signaling pathway, thereby affecting the TGF-β1/Smad signaling pathway. Materials and Methods: This study used a rat model of knee extensor joint immobilization to assess the impact of nicorandil on arthrogenic contracture and joint capsule fibrosis. Sixty Sprague–Dawley rats were divided into control, immobilization, and nicorandil treatment groups with varying durations of immobilization. The extent of arthrogenic contracture was assessed using joint mobility metrics, while joint capsule fibrosis was quantified using Masson staining, α-SMA immunohistochemistry, and H&E staining. The protein expression levels of the TGF-β1/Smad and RhoA/ROCK signaling pathways were analyzed by western blotting. Additionally, lysophosphatidic acid (LPA), a specific ROCK activator, was used to further investigate the involvement of RhoA/ROCK signaling in TGF-β1/Smad pathway modulation. Results: This study revealed a positive correlation between the duration of immobilization and the severity of arthrogenic contracture, joint capsule fibrosis, and inflammatory infiltration. Nicorandil administration effectively reduced these immobilization-induced changes and concurrently inhibited the activation of TGF-β1/Smad and RhoA/ROCK signaling pathways. Application of LPA counteracted the inhibitory effects of nicorandil on these pathways. Conclusion: Nicorandil significantly alleviated arthrogenic contracture and joint capsule fibrosis resulting from knee extension immobilization. The underlying mechanism appears to involve inhibition of the TGF-β1/Smad signaling pathway, mediated by RhoA/ROCK signaling pathway suppression.

ABSTRACT Chicken abdominal fat (AF) is an economically important trait, and many studies have been conducted on genetic selection for AF. However, previous studies have focused on detecting functional chromosome mutations or regions using gene chips. The present study used the specific-locus amplified fragment sequencing (SLAF-seq) technology to perform a genome-wide association study (GWAS) on purebred Wengshang Barred chicken. A total of 1,286,715 single-nucleotide polymorphisms (SNPs) were detected, and 175,211 SNPs were selected as candidate SNPs for genome-wide association analysis using TASSEL general linear models. Two SNPs markers reached genome-wide significance. Of these, rs7943847, rs127627362 were significantly associated with AF at 120 days. These SNPs are close to eight genes (SLC16A6, ARSG, WIPI1, PRKAR1A, FAM20A, ABCA8, ABCA9, CPQ,). These results would enrich the studies on AF and promote the use of Chinese chicken, especially the Wenshang Barred chicken.

OBJECTIVES/GOALS: Whole-genome viral sequencing is vital to inform public health and study evolution. Arboviruses evolve in vectors, reservoir hosts, and humans, and require surveillance at all points. We developed a new rigorous method of sequencing that captures whole viral genomes in field-collected and clinical samples. METHODS/STUDY POPULATION: ClickSeq is a novel method of Next Generation Sequencing (NGS) library synthesis using azido-nucleotides to terminate reverse transcription. The cDNA generated can be ligated to sequencing and indexing primers at room temperature using copper (Cu I) and vitamin C. With this approach, we designed primers located ~250 bp apart along the genomes of the arboviruses Chikungunya 37797, Zika Dakar, Yellow Fever Asibi, Dengue serotype 2, West Nile 385-99, and St. Louis Encephalitis Virus (SLEV) clade II. We tested this method with varying viral titers: lab-infected mosquito pools, field-collected mosquito pools from a Texas West Nile and SLEV outbreak, and patient isolates from a Pakistani CHIKV outbreak. The cDNA was sequenced in the UTMB NGS Core and aligned using bowtie. RESULTS/ANTICIPATED RESULTS: The use of a single protocol to capture whole viral genomes including UTRs for multiple viruses from different sample collection styles is ideal for arboviruses. Primers for multiple viruses were pooled and used to sequence mosquito pools. The Tiled ClickSeq method captured whole viral genomes without the need for host depletion. UTRs were captured even when the viral strain used for primer design differed from the resulting strain. Discreet variants were captured in both the hypervariable nsP3 region and the UTR in the patient isolates from the CHIKV outbreak compared to the 2017 outbreak. Texas WNV and SLEV outbreaks are now defined from the 2020 outbreak and can be further tracked to update public health measures and understand viral evolution. DISCUSSION/SIGNIFICANCE: UTRs impact both human and mosquito fitness, leading to further outbreaks. Tiled ClickSeq aims to capture whole viral genomes with a method and cost that can be implemented by public health researchers to understand disease evolution as it happens to update both public health and basic virology to the effects of evolution on arboviruses.

ABSTRACT The effects of plant essential oil (PEO) on the production performance and immune function of laying hens were evaluated to provide theoretical basis for promoting the natural plant extracts. Eight thousand 1-day-old healthy laying hens were randomly divided into a control group or PEO group, with four replicates per treatment and 1000 hens per replicate. The PEO diet was supplemented with 3g/kg plant extract. Diets were fed for 56 days. The tibia length and keel length were detected on an empty stomach at the end of the trial. Blood samples were collected on the 28th and 56th days to detect the level of C3, C4, IL-1, IL-2, IL-17 and immunoglobulin in the serum. The results showed that, compared with the control, PEO supplementation significantly increased the weight gain rate (WGR) at the 2nd, 4th, 5th and 7th week (p

Abstract This article deals with some existence and Ulam-Hyers-Rassias stability results for a class of functional differential equations involving the Hilfer-Hadamard fractional derivative. An application is made of a Schauder fixed point theorem for the existence of solutions. Next we prove that our problem is generalized Ulam-Hyers-Rassias stable.

In this paper, plasmonic responses of phosphorene in the presence of strain and doping have been systematically investigated. Based on density functional theory, permittivities include both the intraband and interband transitions of electrons have been calculated. Due to the modification of the band structure, significantly higher Drude plasma frequency has been observed along the zigzag direction, other than the armchair direction as in the usual case. The resulting unusual plasmonic responses change their anisotropy, both in the elliptic as well as the hyperbolic regimes. Based on our calculations, positive strain as large as 5% along the zigzag direction can even lead to so-called reversed hyperbolic plasmonic responses. The k -surfaces of the plasmonic modes in extended monolayer have been analytically solved, and it is found that actively switching the topology (between elliptic and hyperbolic regimes) of the plasmonic responses by changing the Fermi level is possible in phosphorene at certain frequencies. In the end, a simple model has been proposed to describe such plasmonic responses in the infrared and the parameters of the model have been listed in tables which can be used directly in calculating the permittivities. Our studies may extend the scope of existing investigations of phosphorene plasmons and lead to band engineering as a way to control plasmons in two-dimensional materials.

Feeding ratio and frequency have substantial effects on food digestion and assimilation in fish, yet few attempts have been made to determine their influences on the bioaccumulation of trace metals. In this study, juvenile Acanthopagrus schlegeli were exposed to either waterborne or dietary cadmium (Cd) at different feeding ratios (0, 2, or 4% body weight d-1 [BW d-1] under waterborne Cd exposure and 1, 2.5, or 4% BW d-1 under dietary Cd exposure) or feeding frequencies (1, 2 or 8 times d-1) for 5 wk. Under waterborne Cd exposure, the fish fed 0% BW d-1 or 8 times d-1 showed significantly lower growth rates and those fed 0% BW d-1 or 1 time d-1 exhibited higher Cd body burdens and Cd uptake rates compared to the other groups. The gut showed a significantly higher waterborne Cd uptake rate than the gills when fish were fasted or fed only 1 time d-1. These results suggest that starvation and low feeding frequency facilitate waterborne Cd uptake. Under dietary Cd exposure, the fish fed 4% BW d-1 or 2 times d-1 grew faster. Cd body burden and Cd assimilation were higher in fish fed 2.5% BW d-1 or 2 times d-1. Dietary Cd retention was positively correlated with feed efficiency, suggesting the utilization of Cd probably coincides with the essential nutrients in fish. Overall, this study demonstrates that different feeding strategies significantly influence waterborne and dietary Cd bioaccumulation in marine fish. Therefore, feeding conditions have to be considered carefully for managing trace metal contamination in marine fish farming.

Myosin, as a type of molecular motor, is mainly involved in muscle contraction. Recently, myosin research has made considerable progress. However, the function of Bombyx mori myosin remains unclear. In this study, we cloned the BmMyosin II essential light chain (BmMyosin II ELC) gene from a cDNA library of silkworm, which had an open reading frame (ORF) of 444 bp encoding 147 amino acids (about 16 kDa). After analyzing their sequences, BmMyosin II ELC was similar to the ELCs of 27 other Myosin II types, which contained EFh domain that bound Ca2+. In addition, 28 sequences had five motifs, motifs 1 and 3 were relatively conserved. We constructed two vectors with BmMyosin to transfect MGC803 or BmN, monolayer wound healing of cells indicated they can promote cell migration successfully. For three fifth instar silkworms, Bm306, BmNB, BmBC8, we mainly analyzed the change of BmMyosin II ELC from transcription and translation after infecting with nucleopolyhedrovirus (BmNPV). We found that gene expression of resistant strains were higher than susceptible strains at 12 h, while the result of the translation level was opposite that of the transcription level. Through in vitro protein interactions, we found BmMyosin II ELC can interact with BmNPV ubiquitin.

Although in vivo studies have shown that low-magnitude, high-frequency (LMHF) vibration (LM: < 1 ×g; HF: 20-90 Hz) exhibits anabolic effects on skeletal homeostasis, the underlying cellular/molecular regulation involved in bone adaptation to LMHF vibration is little known. In this report, we tested the effects of microvibration (magnitude: 0.3 ×g, frequency: 40 Hz, amplitude: ± 50 μm, 30 min/12 h) on proliferation and osteodifferentiation of bone marrow-derived mesenchymal stromal cells (BMSCs) seeded on human bone-derived scaffolds. The scaffolds were prepared by partial demineralisation and deproteinisation. BMSCs were allowed to attach to the scaffolds for 3 days. Morphological study showed that spindle-shaped BMSCs almost completely covered the surface of bone-derived scaffold and these cells expressed higher ALP activity than those cultured on plates. After microvibration treatment, BMSC proliferation was decreased on day 7 and 10; however, numbers of genes and proteins expressed during osteogenesis, including Cbfa1, ALP, collagen I and osteocalcin were greatly increased. ERK1/2 activation was involved in microvibration-induced BMSC osteogenesis. Taken together, this study suggests that bone-derived scaffolds have good biocompatibility and show osteoinductive properties. By increasing the osteogenic lineage commitment of BMSCs and enhancing osteogenic gene expressions, microvibration promotes BMSC differentiation and increase bone formation of BMSCs seeded on bone-derived scaffolds. Moreover, ERK1/2 pathway plays an important role in microvibration-induced osteogenesis in BMSC cellular scaffolds.

We report the discovery of five white dwarf + ultracool dwarf systems identified as common proper motion wide binaries in the Gaia Catalogue of Nearby Stars. The discoveries include a white dwarf + L subdwarf binary, VVV 1256−62AB, a gravitationally bound system located 75.6 |$^{+1.9}_{-1.8}$|  pc away with a projected separation of 1375 |$^{+35}_{-33}$|  au. The primary is a cool DC white dwarf with a hydrogen dominated atmosphere, and has a total age of |$10.5^{+3.3}_{-2.1}$|  Gyr, based on white dwarf model fitting. The secondary is an L subdwarf with a metallicity of [M/H] = |$-0.72^{+0.08}_{-0.10}$| (i.e. [Fe/H] = |$-0.81\pm 0.10$|⁠) and |$T_{\rm eff}$| = 2298 |$^{+45}_{-43}$|  K based on atmospheric model fitting of its optical to near infrared spectrum, and likely has a mass just above the stellar/substellar boundary. The subsolar metallicity of the L subdwarf and the system's total space velocity of 406 km s−1 indicates membership in the Galactic halo, and it has a flat eccentric Galactic orbit passing within 1 kpc of the centre of the Milky Way every |$\sim$| 0.4 Gyr and extending to 15–31 kpc at apogal. VVV 1256−62B is the first L subdwarf to have a well-constrained age, making it an ideal benchmark of metal-poor ultracool dwarf atmospheres and evolution. [ABSTRACT FROM AUTHOR]