Biomedicines

Alcohol-related hepatitis (AH) is plagued with high mortality and difficulty in identifying at-risk patients. The extracellular matrix undergoes significant remodeling during inflammatory liver injury that can be detected in biological fluids and potentially used for mortality prediction. EDTA plasma samples were collected from AH patients (n= 62); Model for End-Stage Liver Disease (MELD) score defined AH severity as moderate (12-20; n=28) and severe (>20; n=34). The peptidome data was collected by high resolution, high mass accuracy UPLC-MS. Univariate and multivariate analyses identified differentially abundant peptides, which were used for Gene Ontology, parent protein matrisomal composition and protease involvement. Machine learning methods were used on patient-specific peptidome and clinical data to develop mortality predictors. Analysis of plasma peptides from AH patients and healthy controls identified over 1,600 significant peptide features corresponding to 130 proteins. These were enriched for ECM fragments in AH samples, likely related to turnover of hepatic-derived proteins. Analysis of moderate versus severe AH peptidomes showed a shift in abundance of peptides from collagen 1A1 and fibrinogen A proteins. The dominant proteases for the AH peptidome spectrum appear to be CAPN1 and MMP12. Increase in hepatic expression of these proteases was orthogonally-validated in RNA-seq data of livers from AH patients. Causal graphical modeling identified four peptides directly linked to 90-day mortality in >90% of the learned graphs. These peptides improved the accuracy of mortality prediction over MELD score and were used to create a clinically applicable mortality prediction assay. A signature based on plasma peptidome is a novel, non-invasive method for prognosis stratification in AH patients. Our results could also lead to new mechanistic and/or surrogate biomarkers to identify new AH mechanisms. Lay summaryWe used degraded proteins found the blood of alcohol-related hepatitis patients to identify new potential mechanisms of injury and to predict 90 day mortality.

BackgroundPancreatic ductal adenocarcinoma (PDAC) is poised to be a leading cause of cancer-related deaths. Despite developing new treatment strategies, patient outcomes have not significantly improved. Chemoresistance has been implicated as a major contributor to ineffective treatments observed with studies suggesting combination therapy targeting multiple pathways. This study explored dysregulated genes in tumours of PDAC patients to identify targets which could be used effectively in combination with conventional therapy against cancer cells. MethodsIn this study, PCR arrays were used for gene expression profiling of tumours obtained from South African PDAC patients to identify key differentially expressed pathways and potentially new therapeutic target genes. SPP1 was selected and RNA interference was used to knock the gene down. Migration and apoptosis assays were used to evaluate the effect of the knockdown, alone and in combination with gemcitabine, on a pancreatic cancer cell line, MIA PaCa-2. Proteomic analysis using SWATH-MS was used to demonstrate potential molecular mechanisms linked to the morphological and phenotypical effects observed with treatment. ResultsWe demonstrated several genes linked to the growth factor and signal transduction signalling pathways, and identified SPP1 as a target. We observed that by combining SPP1 knockdown with conventional chemotherapy, gemcitabine, resulted in a synergistic effect, leading to an enhanced early apoptotic response. A decline of migratory and invasive capabilities of MIA PaCa-2 cells was observed upon subjecting the cancer cells to SPP1 reduction and gemcitabine treatment. Furthermore, proteomic analyses uncovered several pathways that were dysregulated by the combination therapy including both pro-and anti-tumorigenic ones. ConclusionThe study findings indicate that SPP1 could be a potential therapeutic target for PDAC, and the possible synergistic effects observed when SPP1 knockdown was combined with gemcitabine treatment suggest a potential avenue for developing more effective treatments for PDAC while exploring tumour cell adaptation for survival.

Pancreatic ductal adenocarcinoma (PDAC) is a lethal cancer with a growing incidence and mortality despite novel therapeutic strategies. The complement signalling pathway may play diverse roles in PDAC by eliciting an immune response, inducing inflammatory responses, and may elevate pathways linked to chemoresistance. However, their role in the progression of PDAC is not fully understood. In this study, 30 tissues and 34 plasma samples were obtained from a cohort of PDAC patients including controls. Targeted pathway-specific PCR analysis was conducted to determine the gene expression profiles of immune-response-related genes. The circulating levels of complement proteins C3 and C5 were further investigated. Pharmacological inhibition of the complement pathway in MIA PaCa-2 pancreatic cancer cell lines was performed and the effect on cells was assessed by cell proliferation, cell migration, and cell cycle assays. Finally, SWATH-mass spectrometry was performed to identify potential molecular mechanisms during inhibition. The results identified C3 and C5 to be overly expressed in early PDAC compared to later stages. Pharmacological inhibition of the complement pathway led to increased cell growth, proliferation and migration in vitro. Proteomic analysis implicated several proteins such as the mitochondrial and histone proteins, that could play a role in inducing this phenotype. This study helps to further delineate the role of the complement pathway in PDAC progression suggesting a context-dependent function.

This study aimed to evaluate the prognostic value of quantitative analysis of {superscript 1}F-FDG positron emission tomography (PET)/computed tomography (CT) metabolic parameters in patients with pancreatic ductal adenocarcinoma (PDAC) after neoadjuvant chemotherapy (NACT). A retrospective analysis was conducted on the clinical and imaging data of 44 patients with pathologically confirmed PDAC who received NACT. All patients completed standard chemotherapy regimens and underwent {superscript 1}F-FDG PET/CT examinations within 2 weeks before and after chemotherapy. Multiple metabolic parameters of lesions were extracted, their percentage changes were calculated, and the optimal cut-off values for each parameter were determined. Kaplan-Meier survival analysis and Cox proportional hazards regression analysis were applied to explore the prognostic value of the metabolic parameters, and the prognostic stratification performance of PET Response Criteria in Solid Tumors (PERCIST) 1.0 was compared with that of Response Evaluation Criteria in Solid Tumors (RECIST) 1.1. PERCIST 1.0 demonstrated significantly superior prognostic stratification compared with RECIST 1.1. A peak standardized uptake value corrected for lean body mass (SULpeak2) > 3.07 and a percentage change in SULpeak between pre- and post-treatment scans ({Delta}SULpeak%) [≤] 37.66% were identified as independent risk factors for poor prognosis. Furthermore, SUL-related parameters exhibited markedly better predictive efficacy than traditional metabolic parameters such as the standardized uptake value and metabolic tumor volume. Quantitative analysis of {superscript 1}F-FDG PET/CT metabolic parameters can effectively predict prognosis in PDAC after NACT, and PERCIST 1.0 is a more optimal criterion for efficacy and prognostic assessment. A post-NACT SULpeak > 3.07 and {Delta}SULpeak% [≤] 37.66% were core independent indicators for predicting poor prognosis in these patients.

BackgroundThe management of transthyretin amyloid cardiomyopathy (ATTR-CM) has revolved around the scintigraphic diagnosis since the introduction of a specific treatment; however, the equivalency of the bone radiotracers remains unclear. This retrospective monocentric observational study compared [99mTc]Tc-3,3-diphosphono-1,2-propanodicarboxylic acid ([99mTc]Tc-DPD) and [99mTc]Tc-hydroxy-methylene diphosphonate ([99mTc]Tc-HMDP) for ATTR-CM diagnosis. MethodsOne hundred and twenty-nine patients who underwent single photon emission computed tomography (SPECT/CT) after intravenous injection of [99mTc]Tc-DPD or [99mTc]Tc-HMDP for ATTR-CM were included. The patients current visual Perugini grades were retrieved. Regions of interest (ROI) were measured on the heart (H) and on contralateral mediastinum (CM), and H/CM ratios were calculated. ResultsAlthough the distribution of quantitative assessments of heart to contralateral mediastinum ratios is wider with [99mTc]Tc-DPD, suggesting a trend towards improved diagnosis, no difference in Perugini grades was found between [99mTc]Tc-DPD or [99mTc]Tc-HMDP for the diagnosis of ATTR-CM in evocative/non-evocative conditions. There was no difference in ATTR-CM diagnosis between the 2 tracers with a threshold of 1.5 (p-value = 3.316*10-10 for [99mTc]Tc-HMDP and p-value = 2.59*10-15 for [99mTc]Tc-DPD). ConclusionsWe show in our local cohort that [99mTc]Tc-DPD and [99mTc]Tc-HMDP for amyloidosis diagnostic are equivalent for ATTR-CM diagnosis based on the Perugini grading scale. With [99mTc]Tc-DPD, a wider range of the H/CM ratio was noted, which may be considered as a better discrimination of the disease by this bone tracer. Additional research with a final diagnosis of the disease is necessary to evaluate the utility of this quantitative evaluation.

Background and AimsSevere alcoholic hepatitis (SAH) has a high mortality and corticosteroid therapy is effective in 60% patients. Reliable indicators of response to therapy and mortality in SAH are needed. A total of 223 SAH patients, 70 in derivative [50 responders (R) and 20 non-responders (NR)] and 153 in validation cohort [136R, 17NR] were subjected to plasma metabolic/meta-proteomic analysis using UHPLC-HRMS and validated using Machine-Learning (ML). Temporal metabolic changes were assessed using Weighted Metabolome Correlation Network Analysis (WMCNA). Functionality (inflammatory-nature, effect on membrane integrity and glucocorticoid receptor) of non-response indicator was assessed in-vitro on primary healthy neutrophils or mice enterocytes. Baseline plasma metabolomics and meta-proteomics clearly discriminated NR and showed significant increase in urobilinogen (3.6-fold), cholesterol sulfate (6.9-fold), Adenosine monophosphate (4.7-fold) and others (p<0.05, FC>1.5, FDR<0.01). Increase in alpha/beta diversity, biosynthesis of secondary metabolites was a characteristic feature of NR (p<0.05). NR were metabolically inactive however R showed temporal change in the metabolite expression post-corticosteroid therapy (p<0.05). Plasma urobilinogen predicted non-response [AUC=0.94] with a hazard-ratio of 1.5(1.2-1.6) and cut-off >0.07mg/ml segregated non-survivors (p<0.01) and showed >98% accuracy using ML. Plasma urobilinogen directly correlated with circulating bacterial peptides linked to bilirubin to urobilinogen metabolising bacteria (r2>0.7;p<0.05). Urobilinogen induced neutrophil activation, oxidative-stress and pro-inflammatory cytokines (CXCR1, NGAL, NOXO1, NOX4, IL15, TNF and others, p<0.05), promoted corticosteroid resistance by increasing the expression of GR-Beta and trans-repression genes under GR-alpha (inflammatory-NFkB, MAPK-MAP) and reducing GR-alpha, and transactivation (anti-inflammatory) gene levels. Urobilinogen also promoted leaky gut by deregulating intestinal membrane junction proteins. ConclusionPlasma metabolome/meta-proteome can stratify pre-therapy steroid response. Increase in plasma Urobilinogen pedals a vicious cycle of bacterial translocation and increase in inflammation and corticosteroid non-response in SAH patients.

Alzheimers disease (AD) is a multifactorial disorder whose hallmark lesions have been recognized for more than a century, yet its molecular pathobiology remains fragmentary. During our AD research, we discovered a direct mechanistic axis that links the Notch signalling pathway -- a crucial regulator of cell differentiation and proliferation--with the small nuclear RNAs (snRNAs) that drive pre-mRNA splicing. For the first time, our discovery unifies two previously siloed paradigms -- cell signal transduction and RNA splicing, expanding the fundamental knowledge of gene expression and its regulation. The first contribution of the present study is the discovery and validation of the Notch/snRNA negative feedback regulation. More implications include: (1) it reframes splicing-related diseases as dynamic consequences of pathway dysregulation rather than static snRNA-gene mutations; (2) it explains why global splicing abnormalities appear in AD, aging, and cancer even when snRNA genes themselves are not mutated; and (3) it provides an immediate, druggable "splicing rheostat" for any Notch-hyperactive disorder, such as AD.

Doxorubicin is a member of the anthracycline class of chemotherapeutic agents and is among the most effective treatments available up to date. However, its clinical use is significantly limited by severe cardiotoxic effects. The purpose of this study is to investigate the transcriptomic alterations that occur in a rat model of doxorubicin-induced cardiotoxicity. Our results reveal significant dysregulations of cardiac metabolism and provide insights into the molecular mechanisms underlying cardiac damage produced by this treatment. Our analysis revealed that heart tissue recovery following doxorubicin treatment is hindered by hypercholesterolemia exacerbated by transcriptomic-level alteration of the circadian rhythm. This result could help facilitate the discovery of circulating biomarker-based early diagnostic methods for this disease, along with cardioprotective treatment implementation.

BackgroundDelirium following cardiac surgery is common, morbid, and costly, but may be prevented with risk stratification and targeted intervention. Preoperative protein signatures may identify patients at increased risk for worse postoperative outcomes, including delirium. In this study, we aimed to identify plasma protein biomarkers and develop a predictive model for postoperative delirium in older patients undergoing cardiac surgery, while also uncovering possible pathophysiological mechanisms. MethodsSOMAscan analysis of 1,305 proteins in the plasma from 57 older adults undergoing cardiac surgery requiring cardiopulmonary bypass was conducted to define delirium-specific protein signatures at baseline (PREOP) and postoperative day 2 (POD2). Selected proteins were validated in 115 patients using the ELLA multiplex immunoassay platform. Proteins were combined with clinical and demographic variables to build multivariable models that estimate the risk of postoperative delirium and bring light to the underlying pathophysiology. ResultsA total of 115 and 85 proteins from SOMAscan analyses were found altered in delirious patients at PREOP and POD2, respectively (p<0.05). Using four criteria including associations with surgery, delirium, and biological plausibility, 12 biomarker candidates (Tukeys fold change (|tFC|)>1.4, Benjamini-Hochberg (BH)-p<0.01) were selected for ELLA multiplex validation. Eight proteins were significantly altered at PREOP, and seven proteins at POD2 (p<0.05), in patients who developed postoperative delirium compared to non-delirious patients. Statistical analyses of model fit resulted in the selection of a combination of age, sex, and three proteins (angiopoietin-2 (ANGPT2); C-C motif chemokine 5 (CCL5); and metalloproteinase inhibitor 1 (TIMP1); AUC=0.829) as the best performing predictive model for delirium at PREOP. The delirium-associated proteins identified as biomarker candidates are involved with inflammation, glial dysfunction, vascularization, and hemostasis, highlighting the multifactorial pathophysiology of delirium. ConclusionOur study proposes a model of postoperative delirium that includes a combination of older age, female sex, and altered levels of three proteins. Our results support the identification of patients at higher risk of developing postoperative delirium after cardiac surgery and provide insights on the underlying pathophysiology. ClinicalTrials.gov (NCT02546765).

BackgroundNeurologic and neurobehavioural complications are common after cardiac surgery with cardiopulmonary bypass (CPB). Exposure to the artificial bypass surface, conversion to laminar flow and hypothermia likely contribute to systemic inflammation observed after CPB. To ensure adequate systemic oxygenation, the CPB patient is often exposed to supraphysiologic levels of oxygen. Relative to normoxia, perioperative hyperoxia during CPB has not been shown to impact neurocognition in the long-term. Whether this holds true for the immediate post-operative neurocognitive function is the question of this nested case-control study. Methods46 age-and sex-matched subjects, aged [&ge;]65 years, selected for this study were randomized to receive normoxia or hyperoxia during CABG with CPB in the parent trial. Levels of four neuroinflammatory biomarkers (S100B, ENO2, CHI3L1, UCHL1) were measured at baseline and at post-bypass. Baseline neurocognition was established with the Montreal Cognitive Assessment tool and patients were assessed on each post-operative day for delirium using the confusion assessment method. Mediation analyses was conducted for the conditional effect of perioperative oxygen treatment on the occurrence of delirium, assuming mediation effect from change in biomarker levels. Results26 subjects (n = 12) demonstrated delirium. Of the four biomarkers, only S100B levels were differentially abundant post-bypass regardless of treatment (8.18 versus 10.15pg/mL, p value < 0.001). We found significant direct effects of treatment on the occurrence of delirium (effect size = -2.477, p = 0.004). There was no statistically significant mediating effect by S100B. ConclusionWhile perioperative hyperoxia may not be associated with neurocognitive dysfunction in the long-term, its immediate effects may contribute significantly to the occurrence of post-operative delirium. Taken together, our findings suggest a dose-response-time relationship between hyperoxia and neurocognitive function.

Takotsubo syndrome (TTS), an acute cardiac condition characterized by transient wall motion abnormalities mostly of the left ventricle, results in difficulties in diagnosing patients. We set out to present a detailed blood analysis of TTS patients analyzing novel markers to understand the development of TTS. Significant differences in pro-inflammatory cytokine expression patterns, sex steroid and glucocorticoid receptor (GR) expression levels were observed in the TTS patient collective. Remarkably, the measured catecholamine serum concentrations determined from TTS patient blood could be shown to be two orders of magnitude lower than the levels determined from experimentally induced TTS in laboratory animals. Consequently, exposure of endothelial cells and cardiomyocytes in vitro to such catecholamine concentrations did not damage the cellular integrity or function of neither endothelial cells forming the blood brain barrier, endothelial cells derived from myocardium nor cardiomyocytes in vitro. Computational analysis was able to link the identified blood markers, specifically, the pro-inflammatory cytokines and glucocorticoid receptor GR to microRNA (miR) relevant in the ontogeny of TTS (miR-15), and inflammation (miR-21, miR-146a), respectively. Amongst the well-described risk factors of TTS (older age, female gender), inflammaging-related pathways were identified to add to these relevant risk factors or prediagnostic markers of TTS.

Ischemic stroke due to large vessel occlusion results from the blockage of a major cerebral artery by a clot; however, the origins and molecular composition of these clots remain poorly understood. Mechanical thrombectomy has become a standard treatment to remove obstructive clots, providing a unique opportunity to analyze their properties. We previously demonstrated that blood can clot into an amyloid-like form, generating fibrinaloid microclots (2-200 m) that are highly resistant to fibrinolysis. In this study, archived clots from eight ischemic stroke patients with large vessel occlusion were examined, using samples stored in the Walton Centre Clot Bank in Liverpool, UK. All clots exhibited strong, heterogeneous amyloid staining, revealing a pervasive amyloid component. These findings represent a previously unreported characteristic of stroke clots, highlighting the potential for amyloid-targeted therapies to overcome their fibrinolytic resistance and providing a foundational new insight into ischaemic stroke pathophysiology and treatment.

BackgroundBarretts Oesophagus (BO) presents a particular pathological dilemma, in that patients who have no dysplasia within their BO experience a small but significant risk of malignant progression each year. Screening programmes have attempted to reduce the mortality from BO associated oesophageal adenocarcinoma but cannot predict which BO patients will progress to invasive malignancy. We have previously identified the long non coding RNA, OR3A4, is differentially hypomethylated in progressive BO. We aimed to understand its role in BO pathogenicity MethodsThe stable BO cell line CP-A, as well as the oesophageal adenocarcinoma cells line OE-33 was transfected with a lentiviral OR3A4 over-expression vector, and underwent high resolution microscopy, immunofluorescence, RT-qPCR, RNA sequencing, and targeted drug screening with the p38-MAPK inhibitor domipramod to understand the effects of OR3A4 expression on progression. We then compared progressive vs. non-progressive BO samples using quantitative multi-fluorophore (Vectra) immunohistochemistry. ResultsOver-expression of OR3A4 in CP-A lines resulted in a hyperproliferative, dysplastic cellular phenotype, with strong over-expression of MAPK and anti-apoptotic pathways at the RNA and protein level, which was sensitive to the p38-MAPK inhibitor domipramod. Vectra immunohistochemistry demonstrated that progressive BO had reduced visibility associated with a reduction in CD8+ T-cells and CD68+ macrophages and reduced CD4+ T-cells in the stomal compartment. ConclusionThe overexpression of OR3A4, which we have previously shown is associated with progressive BO leads to a proliferative dysplastic cellular phenotype associated with increased, reversible MAPK signalling and loss of immune visibility.

Background and AimLymphatic vessels (LVs) are crucial for maintaining abdominal fluid homeostasis and immunity. In liver cirrhosis, mesenteric LVs (mLVs) are dilated and dysfunctional. Given an established protective role of VEGF-C in LVs, we hypothesized that VEGF-C treatment could improve functions of mLVs in cirrhosis. MethodIn this study, we developed a nanoformulation comprising LV-specific growth-factor, recombinant human VEGF-C(Cys156Ser) protein(E-VEGF-C) and delivered it orally in rat models of liver cirrhosis to target mLVs. Nanoformulation without VEGF-C served as vehicle. Drainage of mLVs was analyzed using tracer dye. Portal and systemic physiological assessments and computed-tomography were performed to measure portal pressures and ascites. Gene expression of mesenteric lymphatic endothelial cells (LyECs) was studied. Immune cell subsets in mesenteric lymph nodes (MLNs) were quantified by flow-cytometry. Gut bacterial translocation to MLNs was examined using GFP-labelled bacteria. ResultsIn cirrhotic rats, mLVs were dilated and leaky with impaired drainage. Treatment with E-VEGF-C induced proliferation of VEGFR3+ mLVs, reduced their diameter and improved functional drainage. Ascites and portal pressures were significantly reduced in E-VEGF-C treated rats compared to vehicle. At molecular level, E-VEGF-C treatment upregulated the expression of cell adhesion and permeability genes (VCAM1, VE-Cad) in LyECs. In MLNs of E-VEGF-C rats, there was an increased percentage of CD8+CD134+ T-cells and decreased CD25+Treg-cells. Bacterial translocation was also limited to MLNs only in E-VEGF-C treated rats with reduced levels of endotoxins in ascites in comparison to vehicle. ConclusionE-VEGF-C treatment ameliorates mesenteric lymph drainage, portal pressure, and strengthens cytotoxic immune responses in MLNs in experimental cirrhosis. It may thus serve as a promising therapy to manage ascites and portal pressure and reduce gut bacterial translocation in patients with cirrhosis. Lay SummaryA human recombinant pro-lymphangiogenic growth factor, VEGF-C, was encapsulated in nanolipocarriers (E-VEGF-C) and orally delivered in rat models of decompensated liver cirrhosis to facilitate its gut lymphatic vessel uptake. E-VEGF-C administration significantly increased mesenteric lymphatic vessel proliferation and improved lymph drainage, attenuating abdominal ascites and portal pressures in the animal models. E-VEGF-C treatment limits bacterial translocation to MLNs only with reduced gut bacterial load and ascitic endotoxins. E-VEGF-C therapy holds the potential to manage ascites and portal pressure and reduce gut bacterial translocation in patients with decompensated cirrhosis. Graphical Abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=80 SRC="FIGDIR/small/20248815v4_ufig1.gif" ALT="Figure 1"> View larger version (26K): org.highwire.dtl.DTLVardef@1f63596org.highwire.dtl.DTLVardef@1381e52org.highwire.dtl.DTLVardef@14ebe05org.highwire.dtl.DTLVardef@9d559a_HPS_FORMAT_FIGEXP M_FIG C_FIG

Medulloblastoma is the most common childhood brain tumor with an unfavorable prognosis and limited options of harmful treatments that are associated with devastating long-term side effects. Therefore, the development of safe, non-invasive and effective therapeutic approaches is required to save the quality of life of young medulloblastoma survivors. We postulated that therapeutic targeting is a solution. Thus, we used a recently designed tumor-targeted bacteriophage (phage)-derived particle, named transmorphic phage/AAV, TPA, to deliver a transgene expressing the tumor necrosis factor alpha (TNF) for targeted systemic therapy of medulloblastoma. This vector was engineered to display the double cyclic RGD4C peptide to selectively target tumors after intravenous administration. Furthermore, the lack of native phage tropism to mammalian cells warrants safe and selective systemic delivery to the tumor microenvironment. In vitro RGD4C.TPA.TNF treatment of human medulloblastoma cells generated efficient and selective TNF expression, subsequently triggering cell death. Combination with the chemotherapeutic drug cisplatin, used clinically against medulloblastoma, resulted in augmented effect through the enhancement of TNF gene expression. Systemic administration of RGD4C.TPA.TNF to mice bearing subcutaneous medulloblastoma xenografts resulted in selective tumor homing of these particles, and consequently targeted tumor expression of TNF, apoptosis, and destruction of the tumor vasculature. Thus, our RGD4C.TPA.TNF particle provides selective and efficient systemic delivery of TNF to medulloblastoma, yielding a potential TNF anti-medulloblastoma therapy while sparing healthy tissues from the systemic toxicity of this cytokine.

ObjectiveCircular RNA(circRNA) is a kind of endogenous non-coding RNA, which may be related to the occurrence and development of cancer. Based on the GEO database, this paper constructs a circRNA as a competitive endogenous RNAs(ceRNAs) that binds with microRNAs (miRNAs) to affect and regulate the expression of target genes. The ceRNA regulatory network based on circRNA-miRNA-mRNA model plays an important role in tumor prognosis and treatment. This paper explores the mechanism of circRNA-related ceRNA regulatory network in gastric cancer. MethodCircRNA, miRNA and mRNA data sets related to gastric cancer were downloaded from The Gene Expression Omnibus (GEO), and the limma package of R software (R 4.2.1 version) was used to identify the differences between gastric cancer tissues and adjacent normal tissues of gastric cancer. DEcircRNA, DEmiRNAs, and DEmRNAs. Based on circBase database, we explored the interactions among circRNA, miRNA and mRNA, and constructed the circRNA-miRNA-mRNA ceRNA network by using Cytoscape_v3.8.0. Then KEGG, GO and survival analysis of ceRNA-related genes were performed. Then, the prognostic data of gastric cancer were extracted from the TCGA database to construct the prognostic subnetwork of gastric cancer. ResultsKEGG analysis of ceRNAs mRNA showed that the pathway was mainly enriched in IL-17 signaling pathway, TNF signaling pathway and so on, which affected the prognosis of gastric cancer. hsa_circ_0055521/hsa-miR-204-5p/FAP, (hsa_circ_0005051, hsa_circ_0007613, hsa_circ_0045602, hsa_circ_0034398, hsa_circ_0006089) /hsa-miR-32-3p/FNDC1 were the ceRNA networks related to the prognosis of gastric cancer collaterals. ConclusionThis study found that IL-17 signaling pathway and TNF signaling pathway may affect the occurrence, development and prognosis of gastric cancer. And hsa_circ_0055521/hsa-miR-204-5p/FAP, (hsa_circ_0005051, hsa_circ_0007613, hsa_circ_0045602, hsa_circ_0034398, hsa_circ_0006089) /hsa-miR-32-3p/FNDC1 two circrNa-based stomachs The cancer ceRNA prognostic network is a new prognostic related ceRNA network for gastric cancer. FNDC1 and FAP may be potential therapeutic targets for gastric cancer.

1.1Brain cancers are ones of most aggressive and difficult to treat cancers. Despite numerous studies of the cellular mechanisms of gliomas, it is difficult to stop tumor growth. A complex genetic and epigenetic nature of many gliomas and poorly known pathways of human neuron precursors maturation suggest turning to big data analysis to find new insights and directions for drug development. We developed in silico molecular models and predicted molecular switches in signaling cascades that maintain multipotency of neuronal precursor cells in diffuse intrinsic pontine glioma (DIPG) driven by the H3K27M mutation and mutations in the TP53 gene. Oncogenes and biomarkers were predicted based on transcriptomics and mutational genomics data from a cohort of 30 patients with DIPG analyzed using Elsevier artificial intelligence methods and a collection of manually curated cancer hallmark pathways. The molecular models of DIPG with mutations in TP53 and histone 3 gene describe the mechanism of oligodendrocyte dedifferentiation due to activation of transcriptional factors OLIG2, SOX2 and POU5F1, epithelial-to-mesenchymal transition via strong EGFR and TGFR signaling, enhanced cell response to hypoxia via HIF1A signaling, and enhanced angiogenesis by VEGFA overexpression. Using in silico analysis, we identified drugs capable of inhibiting mutant TP53: vorinostat, cisplatin, paclitaxel, and statins were top ranked drugs. The predicted drugs and oncogenes had individual patient-level differences that can be visualized with created DIPG model and may be useful for future research in the field of personalized medicine.

Increase of infections caused by microorganisms resistant to conventional antibiotics is a health problem in Brazil and worldwide. The search for new molecules capable of inhibiting the growth of pathogens is a challenge for researchers, who find in venoms a rich source of biomolecules, including antimicrobial peptides (AMPs). The Brazilian scorpion, Tityus serrulatus, is one of the species that cause serious accidents; its venom is rich in neurotoxins that have been well characterized, highlighting their activities on channels (especially sodium and potassium). In this study we identifified and characterized one AMPs in T. serrulatus venom. After milking, the venom was fractioned by high performance liquid chromatography and the fractions were tested by liquid growth inhibition assay, the minimum inhibitory concentration (MIC) against Escherichia coli, Micrococcus luteus, Candida albicans and Aspergillus niger. The fraction identified with antimicrobial activity was analyzed by electrophoresis and mass spectrometry and this AMP (with molecular mass 6.882 kDa) has a similar amino acid sequence to TsTX-{kappa} beta, a neurotoxin that acts on ion channels. The TsTX-{kappa} beta in this study was identified by de novo sequencing. This peptide showed activity against all microorganisms tested. At high concentrations, this peptide, showed hemolytic activity against human erythrocytes. This is a new function described for this peptide, the identification of antimicrobial activity in a neurotoxin already known. Key ContributionMultifunction: antimicrobial and hemolytic activity associated to TsTX-{kappa} beta, a neurotoxin that acts on potassium channels.

Oxymatrine (OMT), is a natural quinoxaline alkaloid from the traditional Chinese medicine herb and has been shown to exhibit anticancer properties on various types of cancer cells. Poorly differentiated gastric adenocarcinoma is a common malignancy of gastric cancer that is more aggressive and has a poor prognosis. In the present study, we investigate the effects of Slit2/Robo1 signals in poorly differentiated gastric adenocarcinoma and adjacent tissues, and the anticancer properties of OMT on human poorly differentiated gastric adenocarcinoma BGC-823 cells and evaluate their underlying mechanisms. The expression levels of Slit2 and Robo1 proteins were measured in 20 pairs of human poorly differentiated gastric adenocarcinoma tissues and adjacent normal tissues using western blot. The expression of apoptosis related proteins and autophagy-related proteins was detected by western blot. The cells viability was detected by CCK-8 assay. The migration of BGC-823 cells was detected by transwell experiments. The expression of related proteins was detected by western blot. The result shows that Slit2 and Robo1 are significantly increased in poorly differentiated gastric adenocarcinoma. The apoptosis and autophagy are inhibited in poorly differentiated gastric adenocarcinoma. OMT inhibits the growth and migration of BGC-823 cells in vitro. OMT inhibits the activation of Slit2/Robo1 signals and induces apoptosis and autophagy in BGC-823 cells. These findings suggest that the antitumor effects of OMT may be the result of inhibition of cell growth and migration, and inhibits the activation of Slit2/Robo1 signals pathway and induces apoptosis and autophagy. OMT may represent a novel anticancer therapy for the treatment of poorly differentiated gastric adenocarcinoma.

AimsThere is a bidirectional link between hypertension (HT) and cellular senescence of endothelial cells (ECs). However, the mechanisms underlying EC senescence in patients with HT are not yet fully understood. Methods and ResultsWe analyzed serum from 71 patients with primary HT and compared it to serum from 25 healthy donors to assess its effects on EC biology, including biomarkers, signaling pathways, and cellular senescence effectors. Our findings revealed that exposing ECs to serum from HT patients (20% for 72 h) impaired cell viability while enhancing proliferation, migration, and tubulogenesis. This effect is accompanied by increased expression of HIF-1. Additionally, HT serum potentiated the expression of the senescence marker SA-{beta}-Gal, shortened telomeres, and up-regulated cell-cycle inhibitors p16, p21, and p53. Regarding the signaling pathways, HT serum activated ERK1/2, p38 MAPK, AP-1/c-jun, and Notch1. Indices of oxidative stress in ECs treated with HT serum also increased, as indicated by elevated production of superoxides, activation of antioxidants (SOD, CAT), and accumulation of oxidized DNA, proteins, and lipids. Furthermore, mitochondria in these cells displayed decreased inner membrane potential and increased biogenesis, likely due to enhanced activity of PGC-1. The activity of respiratory chain enzymes, including cytochrome c oxidase and NADH dehydrogenase, was also elevated. When HT serum-treated ECs were pre-incubated with the ROS scavenger PBN, the activity of SA-{beta}-Gal decreased. A similar reduction in SA-{beta}-Gal activity was observed when HT serum, which contained elevated levels of TGF-{beta}1, was pre-incubated with a TGF-{beta}1-neutralizing antibody. Importantly, exogenous TGF-{beta}1, administered at a dose corresponding to its concentration in HT serum, induced senescence in ECs. ConclusionsOur results indicate that serum from HT patients promotes senescence in ECs through mechanisms related to TGF-{beta}1 and oxidative stress signaling.