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A Temporal Phospho-acetylome Atlas of Human Myogenesis Identifies coordinated Post-Translational Regulation.

Skeletal muscle differentiation depends on precise temporal regulation of protein modifications. To define how phosphorylation and lysine acetylation change during this process, we applied tandem mass tag (TMT)-based quantitative proteomics to human myoblasts sampled at six stages spanning proliferation, induction of differentiation, and early myotube formation. Using sequential enrichment of phosphorylated and acetylated peptides, high-pH fractionation, and high-resolution mass spectrometry, we identified more than 22,000 modified peptides and quantified their temporal behavior after correcti

Osteopontin Promotes Lung Cancer Growth through NF-κB-Dependent Upregulation of PD-L1 and Suppression of Anti-Tumor Immunity.

BACKGROUND: Resistance to immune checkpoint blockade (ICB) remains a major clinical challenge in lung cancer. Osteopontin (OPN) is frequently overexpressed in lung tumors and correlates with poor prognosis, yet its mechanistic role in shaping the immunosuppressive tumor microenvironment remains incompletely defined. METHODS: Using OPN-overexpressing and p65-manipulated lung cancer models, combined with in vivo anti-PD-L1 therapy, we evaluated tumor growth, NF-κB activation, PD-L1 expression, and intratumoral immune composition. Biochemical and molecular assays, including nuclear-cytoplasmic f

Antibacterial activity of plant essential oils against indigenously characterized methicillin-resistant Staphylococcus aureus (MRSA).

Plant essential oils were evaluated for antimicrobial activity against Methicillin-resistant Staphylococcus aureus (MRSA). The isolates (n=03) were procured from Institute of Microbiology, UVAS Lahore, Pakistan. After biochemical and 16S rRNA gene-based PCR characterization, accession numbers were retrieved from NCBI i.e. MW344063.1, MW344064.1 and MW344065.1. These isolates exhibited molecular positivity by multiplex PCR for mecA, coa and eta toxin genes. Moreover, these isolates exhibited resistance to cefoxitin, ampicillin, amoxicillin, penicillin, amoxicillin clavulanate, ciprofloxacin, er

Acquired Immunity Is Not Essential for Radiation-Induced Heart Dysfunction but Exerts a Complex Impact on Injury.

While radiation therapy (RT) can improve cancer outcomes, it can lead to radiation-induced heart dysfunction (RIHD) in patients with thoracic tumors. This study examines the role of adaptive immune cells in RIHD. In Salt-Sensitive (SS) rats, image-guided whole-heart RT increased cardiac T-cell infiltration. We analyzed the functional requirement for these cells in RIHD using a genetic model of T- and B-cell deficiency (interleukin-2 receptor gamma chain knockout (IL2RG-/-)) and observed a complex role for these cells. Surprisingly, while IL2RG deficiency conferred protection from cardiac hyper

Norstictic Acid Inhibits Breast Cancer Cell Proliferation, Migration, Invasion, and In Vivo Invasive Growth Through Targeting C-Met.

Breast cancer is a major health problem affecting the female population worldwide. The triple-negative breast cancers (TNBCs) are characterized by malignant phenotypes, worse patient outcomes, poorest prognosis, and highest mortality rates. The proto-oncogenic receptor tyrosine kinase c-Met is usually dysregulated in TNBCs, contributing to their oncogenesis, tumor progression, and aggressive cellular invasiveness that is strongly linked to tumor metastasis. Therefore, c-Met is proposed as a promising candidate target for the control of TNBCs. Lichens-derived metabolites are characterized by th

EphA4 is localized in clathrin-coated and synaptic vesicles in adult mouse brain.

EphA4, a receptor tyrosine kinase, is expressed in various pre-, post- and peri-synaptic organelles and implicated in the regulation of morphological and physiological properties of synapses. It regulates synaptic plasticity by acting as a binding partner for glial ephrin-A3 and possibly other pre- or post-synaptic ephrins. Now, its trafficking mechanisms remain unknown. In this study, we examine the association of EphA4 with transport, clathrin-coated and synaptic vesicles using cell fractionation, vesicle immunoisolation and electron microscopy. EphA4 was found in highly purified fractions o…

Assembly of cyclic enterobacterial common antigen in Escherichia coli K-12.

We describe here the purification and quantification of a water-soluble cyclic form of enterobacterial common antigen (ECA(CYC)) from Escher … s from overnight cultures contained approximately 2 microg ECA(CYC) per milligram (dry weight), and cell fractionation studies revealed that these molecules were localized exclusively in the periplasm. The synthesis and assembly of ECA(CYC) were found to require the wzxE and wzyE genes of the wec gene cluster. These genes encode proteins involved in the transmembran …

Dose rate effectiveness and potentially lethal damage repair in normal and double-strand break repair deficient murine cells by gamma-rays and 5-fluorouracil.

SCID (severe combined immunodeficiency) fibroblasts established from C.B 17-scid/scid embryos showed higher sensitivity to high (1.105 Gy/min) and low (0.00069 Gy/min) dose rate gamma-rays and also to 5-fluorouracil, a cancer sedative producing double-strand breaks, than wildtype cells from C.B17- +/+ embryos. Furthermore, SCID cells were deficient in repairing DNA damage induced by high dose rate gamma-rays even after dose fractionation and after 24 h recovery periods, while wildtype cells showed an apparent repair ability on DNA damage after these gamma-ray exposures. This is the first repor

ATPases of ciliary epithelium: cellular and subcellular distribution and probable role in secretion of aqueous humor.

The distribution of ion-stimulated ATPases of the ciliary epithelium has been examined in tissues from bovine and rabbit eyes. In homogenates of tissues from both species, both Na,K- and anion-stimulated enzyme activities were found, but no K,H-stimulated activity was detected. The anion ATPase had a broad specificity for a number of anions, and was strongly inhibited by thiocyanate. Following separation of pigmented (outer) and non-pigmented (inner) layers of the bovine ciliary epithelium and isolation of the two cell types on density gradients, higher activities of both Na,K- and anion ATPas

Different kinetics of senescence in human fibroblasts and peritoneal mesothelial cells.

Senescence has been reported for a wide variety of human cell types. In cultures of human fibroblasts the process is due to a percentage of the cells becoming senescent at each passage rather than all the cells entering senescence simultaneously at the end of the life span. By measuring the percentage of fibroblasts which are still cycling at each passage, a rate of decline in the growth fraction, which mirrors the rate of senescence, can be obtained. However, such an analysis has never been undertaken in multiple cell types using the same method to identify cycling cells. It is thus unknown i

Activation of cytotoxic cells by syngeneic prostate antigens in experimental autoimmune vesiculo-prostatitis.

In our experimental model of autoimmune vesiculo-prostatitis, obtained by immunization with syngeneic male accessory glands (MAG) and complete Freund's adjuvant, the presence of specific autoreactive cells with cytotoxic activity against prostate antigens was studied. The specific cytotoxicity generated in MAG immunized rats was tested using 51Cr labelled syngeneic prostate cells or labelled chicken erythrocytes coated with specific antigens (MAG homogenate or chromatographic fractions from prostate homogenate) which were used as target cells in a medium free of complement. The addition of spl

Immunological characterization of non-Hodgkin's lymphoma: enrichment of neoplastic cells from lymphoid tissues and blood.

Cell suspensions prepared from lymph nodes, spleen or peripheral blood of patients with non-Hodgkin's lymphoma (NHL) often contain a high percentage of residual nonmalignant cellular elements. By E-rosette sedimentation, it was possible to enrich such suspensions from patients with various types of lymphoma for malignant cells. In patients with a B- or non-B/non-T-cell lymphoma, the neoplastic cells were found in the non-T fraction. The capacity to respond to stimulation by various stimuli was then confined to the T-cell fraction, which contained the residual normal T-cells. In patients with T

Integrated Phytochemical, Immunological, and Network Pharmacology Investigation of Anti-Inflammatory Lignans From Zanthoxylum armatum Fruits.

In the present study, bioactive lignans from Zanthoxylum armatum fruits have been investigated for their anti-inflammatory properties and evaluated using dendritic cell-based immunological assays and network pharmacology. The ethyl acetate fraction was subjected to bioassay-guided fractionation, chromatography, and HPLC to isolate pure compounds, characterised by NMR, LC-QTOF-MS, and single-crystal x-ray diffraction. Biological activities were evaluated using DPPH/ABTS radical scavenging assays, albumin denaturation and haemolysis inhibition assays, and flow cytometric analysis of IL-12p40/70

Dynamics of Aspartate Isomerization and Its Cellular Implications: Evidence for Isoaspartate Formation in ER-Folded Proteins and Stress-Responsive Regulation of PCMT1.

Spontaneous isomerization of aspartate and deamidation of asparagine residues into isoaspartate (isoAsp) constitute major non-enzymatic post-translational modifications that alter protein structure, stability, and turnover. The repair enzyme protein L-isoaspartate O-methyltransferase (PCMT1) catalyzes methylation of isoAsp residues, thereby preventing their accumulation and preserving proteome integrity. Although PCMT1 has been studied extensively in cytoplasm and nucleus, its relationship to endoplasmic reticulum (ER) proteostasis remains poorly understood. Here, we investigated dynamics of a

Ядерная деятельность и взаимодействие белка NS5 ник-Борна энцефалита.

Орфолавирусы являются вирусами РНК, вызывающими значительные заболевания у людей, одомашненных животных и животных. Их протеин NS5 играет центральную роль в репликации вирусов, действуя как в качестве полимеразы РНК, зависимой от РНК, так и в качестве метилпереносазы, одновременно модифицируя клеточные процессы, включая интерферонную реакцию. Несмотря на то, что вирусное воспроизведение носит цитоплазмический характер, белок NS5 из нескольких переносимых комарами ортофлавирных циклов между цитоплазмой и ядром инфицированных человеческих клеток. Однако ядерная локализация и функция NS5 переноси

Совпадение в аннотации оригинала
Orthoflaviviruses are RNA viruses responsible for significant diseases in humans, domesticated animals and wildlife. Their NS5 protein is ce … , such as tick-borne encephalitis virus (TBEV), remained poorly understood. Microscopy analysis and cell fractionation revealed that the NS5 protein of TBEV localized to both the cytoplasm and nucleoplasm of infected cells. Mutagenesis studies identified critical residues required for its nuclear targeting. Mutating these residues in a TBEV replicon abolished viral …

Profilin-2 promotes tumour aggressiveness in oral squamous cell carcinoma via HDAC1 modulation: implications for EMT and targeted therapy.

BACKGROUND: Profilin-2 (PFN2) is implicated in cancer metastasis, yet its significance in oral squamous cell carcinoma (OSCC) is unclear. METHODS: We quantified PFN2 mRNA and protein in 236 OSCC tumours using qRT-PCR and immunohistochemistry, correlating findings with clinicopathology and survival. Gain- and loss-of-function studies were performed in OSCC cell lines and xenograft mice. RNA sequencing with gene-set enrichment, subcellular fractionation, and immunofluorescence delineated PFN2-dependent pathways. Sensitivity to the histone deacetylase (HDAC) inhibitor SAHA was assessed in vitro.

Ovomucoid hydrolysates produced by pepsin stimulate immune activity of RAW 264.7 macrophages via the MAPK/NF-κB pathway.

Ovomucoid (OVM) was hydrolyzed by proteolytic enzymes to increase immune-enhancing effects for health. Bromelain, Neutrase®, papain, and pepsin were used to hydrolysis and immunostimulatory properties of OVM hydrolysates were evaluated in RAW 264.7 macrophages in this study. Among the OVM hydrolysates, pepsin hydrolysate (OMPH) produced peptides with 10-20 kDa molecular weight, as confirmed by SDS-PAGE. In RAW 264.7 macrophages, OVM hydrolysates at 500 μg/mL or less exhibited high cell viability, exceeding 80 %. In the Griess assay, OMPH produced higher levels of nitric oxide (NO) compared to

RNA-Seq Analysis of MCF-7 Breast Cancer Cells Treated with Methyl Gallate Isolated from the Rhizomes of Nymphaea Odorata L. Shows Upregulation of Apoptosis, Autophagy, and Unfolded Protein Canonical Pathways.

The effects of a methanol extract of Nymphaea odorata (MeNO) rhizomes, its fractions and the active compound (methyl gallate, MeG) were investigated in estrogen receptor-positive (ER+) breast cancer cell lines MCF-7 and T47-D:A18, as well as ER-negative line SKBr3. Cell viability and cytotoxicity were determined using CellTiter-Glo® 2.0 assays at concentrations ranging from 1 to 100 μg/mL. Caspase activity and apoptosis were determined using Caspase-Glo® 3/7, Caspase-Glo® 8, and ApoTox-Glo™ triplex assays, as well as qPCR. Total RNA was isolated from MCF-7 cells treated with MeG. RNA-seq libra

YTHDF3-induced degradation of P4HA2 mRNA inhibits glycolysis in papillary thyroid cancer through Hippo signaling pathway.

BACKGROUND: Prolyl-4-hydroxylase-A2 (P4HA2) is a pivotal enzyme involved in the regulation of tumorigenesis and progression. However, the precise biological roles and potential functions of P4HA2 in papillary thyroid cancer (PTC) remain poorly elucidated. METHODS: Gain-of-function and loss-of-function approaches were employed to investigate the underlying biological effects of P4HA2 on PTC cell proliferation and metastasis both in vitro and in vivo. Furthermore, RIP assay, MeRIP assay, polysome fractionation, dual luciferase reporter assay, LC-MS/MS, and rescue experiments were conducted to e

GW4064 inhibits migration and invasion through cathepsin B and MMP2 downregulation in human bladder cancer.

The ability of bladder cancer to invade and metastasize often leads to poor prognosis in bladder cancer patients. The aim of this study was to evaluate the effect of the farnesoid X receptor (FXR) agonist GW4064 on the migration and invasion of human bladder cancer cells. Long-term exposure to GW4064 decreased the colony formation of RT4 and T24 cells. The wound healing migration assay revealed an inhibitory effect of GW4064 on both of these bladder cancer cell lines. In addition, integrin β3 expression and myosin light chain phosphorylation were decreased after GW4064 treatment. Immunocytoche