Alterations In Protein Levels Research Articles

Potential Functions and Causal Associations of GNLY in Primary Open-Angle Glaucoma: Integration of Blood-Derived Proteome, Transcriptome, and Experimental Verification.

Genome-wide association studies (GWAS) have identified multiple genetic loci associated with primary open-angle glaucoma (POAG). However, the mechanisms by which these loci contribute to POAG progression remain unclear. This study aimed to identify potential causative genes involved in the development of POAG. We utilized multi-dimensional high-throughput data, integrating proteome-wide association study(PWAS), transcriptome-wide association study (TWAS), and summary data-based Mendelian randomization (SMR) analysis. This approach enabled the identification of genes influencing POAG risk by affecting gene expression and protein concentrations in the bloodstream. The key gene was validated through enzyme-linked immunosorbent assay (ELISA) analysis. PWAS identified 86 genes associated with altered blood protein levels in POAG patients. Of these, eight genes (SFTPD, CSK, COL18A1, TCN2, GZMK, RAB2A, TEK, and GNLY) were identified as likely causative for POAG (P SMR < 0.05). TWAS revealed that GNLY was significantly associated with POAG at the gene expression level. GNLY-interacting genes were found to play roles in immune dysregulation, inflammation, and apoptosis. Clinical and cell-based validation confirmed reduced GNLY expression in POAG groups. This study reveals GNLY as a significant potential therapeutic target for managing primary open-angle glaucoma.

SP140 represses specific loci by recruiting polycomb repressive complex 2 and NuRD complex

Abstract SP140, a lymphocytic-restricted protein, is an epigenetic reader working as a corepressor of genes implicated in inflammation and orchestrating macrophage transcriptional programs to maintain cellular identity. Reduced SP140 expression is associated both to autoimmune diseases and blood cancers. However, the molecular mechanisms that link SP140 altered protein levels to detrimental effects on the immune response and cellular growth, as well as the interactors through which SP140 promotes gene silencing, remain elusive. In this work, we have applied a multi-omics approach (i.e. interactomics, ChIP-seq and proteomics) in two Burkitt lymphoma cell lines to identify both interactors and target genes of endogenous SP140. We found that SP140 interacts with the PRC2 and NuRD complexes, and we showed that these interactions are functional as SP140 directs H3K27me3 deposition and NuRD binding on a set of target genes implicated in cellular growth and leukemia progression.

Plasma acute phase proteins as potential predictors of intra-amniotic inflammation and infection in preterm premature rupture of membranes.

We aimed to investigate the potential of altered levels of various acute phase proteins (APPs) in the plasma, either used alone or in combination with ultrasound-, clinical-, and conventional blood-based tests, for predicting the risk of intra-amniotic inflammation (IAI), microbial invasion of the amniotic cavity (MIAC), histologic chorioamnionitis (HCA), and funisitis in women with preterm premature rupture of membranes (PPROM). A total of 195 consecutive pregnancies involving singleton women with PPROM (at 23 + 0-34 + 0 weeks) who underwent amniocentesis and from whom plasma samples were obtained at amniocentesis were retrospectively included in this study. Amniotic fluid (AF) was cultured to assess the MIAC and analyzed for interleukin (IL)-6 levels to define IAI (AF IL-6 level of ≥2.6 ng/mL). The plasma concentrations of hepcidin, mannose-binding lectin (MBL), pentraxin-2, retinol-binding protein 4 (RBP4), serum amyloid A1 (SAA1), and serpin A1 were determined using ELISA. Ultrasonographic cervical length (CL), neutrophil-to-lymphocyte ratio (NLR), and C-reactive protein levels were measured. IAI/MIAC was defined as IAI, MIAC, or both. Multivariate logistic regression analyses showed the following: (1) elevated plasma levels of hepcidin and SAA1 and decreased levels of RBP4 in the plasma were independently associated with IAI/MIAC and (2) decreased plasma RBP4 levels were independently associated with funisitis; however, (3) none of the plasma APPs investigated were associated with acute HCA when adjusted for baseline covariates. Using stepwise regression analysis, noninvasive prediction models comprising plasma RBP4 levels, CL, NLR, and gestational age at sampling were proposed, which provided a good prediction of IAI/MIAC and funisitis (area under the curve: 0.80 and 0.72, respectively). Hepcidin, RBP4, and SAA1 were identified as potential APP biomarkers in the plasma predictive of IAI/MIAC or funisitis in patients with PPROM. In particular, combination of these APP biomarkers with ultrasound-, clinical-, and conventional blood-based markers can significantly support the diagnosis of IAI/MIAC and funisitis.

Dynamic changes and prognostic value of glutathione S-transferase alpha in mild cognitive impairment and Alzheimer’s disease

ObjectivesGlutathione S-transferase alpha (GSTα) is an important antioxidant enzyme closely associated with the onset and progression of neurodegenerative diseases. The alterations in GSTα protein levels associated with Alzheimer’s disease and their impact on cognitive abilities remain unclear. Thus, investigating the fluctuations of GSTα protein levels in mild cognitive impairment (MCI) and Alzheimer’s disease (AD) is essential.MethodsDATA were enrolled from the Alzheimer’s Disease Neuroimaging Initiative (ADNI) database, and we studied healthy individuals (as controls, a total of 54), patients with mild cognitive impairment (345), and patients with Alzheimer’s disease (96) A one-year follow-up was conducted to collect data on the dynamic changes of GSTα protein levels in plasma and primary information data, and to analyze the correlation between the changes in GSTα protein levels before and after the follow-up and cognitive function and its predictive value.ResultsPlasma GSTα protein levels were significantly lower in the AD group than in the CN group (0.94 vs1.05, p = 0.04) and the MCI group (0.94 vs1.09, p &amp;lt; 0.001). Plasma GSTα protein level changes were positively correlated with altered MMSE levels in MCI and AD patients (r = 0.09, p = 0.04). The AUC (95% CI) of the area under the prediction curve of plasma GSTα protein levels for MCI was 0.63 (0.54–0.71), p = 0.02, and the AUC (95% CI) of the area under the prediction curve of plasma GSTα protein levels for AD was 0.74 (0.69–0.80), p &amp;lt; 0.001. At the same time, we plotted ROC curves for the difference in the change of plasma GSTα protein levels after 1 year of follow-up. The results showed that the AUC (95% CI) of the area under the prediction curve of plasma GSTα protein levels change for MCI was 0.76 (0.696–0.83), p &amp;lt; 0.001, and the AUC (95% CI) of the area under the prediction curve of plasma GSTα protein levels change for AD was 0.75 (0.69–0.80), p &amp;lt; 0.001.ConclusionThe findings of the study indicated notable differences in GSTα protein levels among patients with MCI and those with AD after a one-year follow-up period. Furthermore, a positive correlation was observed between changes in GST αprotein levels and the decline in both baseline and cognitive function. This suggests that GSTα protein could potentially act as a biomarker for both MCI and AD, offering fresh insights for early detection and intervention strategies.

Serum Levels of Lead and Selected Acute Phase Proteins in Patients with Substance Use Disorders

Background: Chronic inflammation, immune system dysfunction and elevation in the levels of toxic metals such as lead (Pb) are common observations in patients with substance use disorders (SUD). However, there is little information on the acute phase profile of SUD patients with different serum levels of Pb. Therefore, serum levels of selected acute phase proteins in SUD patients with different serum levels of Pb were determined in this study. Methods: A total of 84 adults consisting of 45 patients with SUDs and 39 controls were enrolled into this case-control study. Serum levels of Pb, albumin and C-reactive protein (CRP) were determined using Atomic Absorption Spectrophotometer, bromocresol green (BCG) colorimetric method, and ELISA, respectively. Thereafter, CRP-albumin ratio (CAR) was calculated as appropriate. Results: Serum levels of Pb and albumin were significantly higher in patients with SUDs compared with the controls. Considering variation in acute phase proteins based on Pb level, the serum level of albumin was significantly lower, while the serum CRP level and CAR were slightly higher in patients with SUDs whose Pb level was higher than 5 µg/dL compared with patients whose Pb level was ≤5 µg/dL. No significant differences were observed in the levels of Pb, albumin, CRP, CAR in patients with SUDs who abuse single substance compared with those who abuse multiple substances. Conclusion: SUD is associated with increased serum levels of Pb and albumin, and alteration in the serum levels of acute phase proteins appears to be influenced by the serum Pb level. Therefore, there is the need for routine measurement of Pb level in patients with SUDs as they could benefit from therapeutic interventions involving chelation of Pb which could prevent disordered acute phase responses in the patients and facilitate optimal response to antipsychotics.

Pulsed Red Photobiomodulation Boosts the Inhibition of Oxytocin-Induced Primary Dysmenorrhea in Mice bySuppressing Oxidative Stress and Inflammation.

Increasing evidence has underscored the pivotal role of red photobiomodulation (R-PBM) in analgesic and anti-inflammatory processes; nonetheless, research concerning the effects of pulsed wave on primary dysmenorrhea (PD) remains sparse. This study found that pulsed R-PBM significantly diminished pain responses and levels of PGF2α/PGE2, mitigated uterine swelling, augmented antioxidant capacity, and lowered MDA concentrations, which outperformed continuous wave at the same average irradiance. Furthermore, PW treatment substantially reduced ROS levels and enhanced cell viability in PGF2α induced HUSM cells. NOS levels, especially iNOS, were markedly diminished in the uteri of PD mice, accompanied by significant alterations in inflammation-related genes (Jun, Fos, IL1rn, IL17b) and protein levels, along with pronounced downregulation of calcium ion concentrations after pulsed R-PBM intervention. These findings indicated that pulsed R-PBM may mitigate pain by modulating ROS and NO/NOS, mediated oxidative stress and inflammatory responses. Consequently, pulsed R-PBM emerges as a promising therapeutic strategy for PD.

Development of an sRNA-mediated conditional knockdown system for Chlamydia trachomatis.

We describe a new Chlamydia trachomatis protein depletion method that uses an engineered small RNA (sRNA) to inhibit translation of a target gene. In proof-of-principle experiments, we induced functional knockdown of IncA, a fusion-mediating inclusion membrane protein, as shown with Western blots, loss of IncA staining at the inclusion membrane, and production of multiple chlamydial inclusions within an infected cell. These effects were titratable and reversible. To test for polar effects, we separately targeted the inclusion membrane proteins IncE and IncG, which are expressed from the incDEFG operon. Knockdown of IncE caused loss of IncE and its interacting host protein SNX6 at the inclusion membrane, without affecting IncG protein levels. Similarly, IncG knockdown significantly reduced IncG levels and prevented recruitment of its interacting host protein 14-3-3β, without altering IncE protein levels. These data provide the first genetic evidence that IncE and IncG are necessary for the recruitment of SNX6 and 14-3-3β, respectively, demonstrating the value of this knockdown approach. We also successfully depleted the major chlamydial surface protein, major outer membrane protein (MOMP), which is encoded by a likely essential gene that has not been previously disrupted or knocked down. MOMP knockdown caused severe defects in bacterial morphology and progeny production. Thus, our sRNA-based approach has broad potential as a conditional knockdown method for studying the function of C. trachomatis genes, including essential genes and genes in an operon.IMPORTANCEWe describe a new method to reduce protein levels of a selected gene in the pathogenic bacterium Chlamydia trachomatis. This approach utilizes an engineered small RNA (sRNA) to inhibit translation of the mRNA for a target gene and produced inducible and reversible protein knockdown. Our method successfully knocked down four proteins, including a likely essential gene and individual genes in an operon, without altering protein levels of a neighboring gene. This conditional knockdown method will be useful for studying the function of genes in Chlamydia. It also has the potential to be applied to other obligate intracellular bacteria, including Rickettsia and Coxiella.

Development of a novel NIR-II fluorescence probe for monitoring serum albumin fluctuation in cerebra neurotoxicity induced by manganese exposure.

Manganese is essential for various biological functions; however, excessive exposure can lead to significant health risks, particularly brain neurotoxicity. Understanding manganese-induced alterations in brain serum protein levels and brain function is crucial for elucidating the mechanisms underlying manganese neurotoxicity. To address this, we developed a novel NIR-II fluorescent probe, RSM, characterized by robust binding to serum albumin and high sensitivity. Using RSM, we observed that heightened BSA uptake in cells exposed to elevated manganese concentrations relative to those exposed to lower levels. Furthermore, we successfully detected changes in serum albumin levels induced by manganese neurotoxicity in brain tissue through in situ NIR-II fluorescence imaging. Our findings establish an association between augmented manganese-induced neurotoxicity and elevated serum albumin content in the brain. This work provides a valuable tool for further investigating the mechanisms of toxic molecules.

Tear fluid reflects the altered protein expressions of Alzheimer's disease patients in proteins involved in protein repair and clearance system or the regulation of cytoskeleton.

New biomarkers that improve diagnosis of Alzheimer's disease (AD) are warranted. Tear fluid (TF) containing variety of proteins that reflect pathophysiological changes of systemic diseases makes TF proteins potential biomarker candidates for AD. We investigated the expression levels of TF proteins in persons with mild AD and cognitively healthy controls (CO) to find out if altered proteins may link to the AD pathophysiology. We analyzed the data of the 53 study participants (34 COs, mean age 71 and Mini-Mental State Examination (MMSE) 28.9 ± 1.4 and 19 persons with AD, CDR 0.5-1, mean age 71 and MMSE 23.8 ± 2.8). All went through neurological status examination, cognitive tests, and ophthalmological examination. TF was collected using Schirmer strips. The TF protein content was evaluated via mass spectrometry-based proteomics and label-free quantification. Eleven proteins having a role either in protein repair and clearance system, or regulation of cytoskeleton, showed altered expression in AD group compared to CO group. Seven of them were significantly (p ≤ 0.05) upregulated (Sti1, Twf1, Myl6, Otub1, Pls1 and Caza1) or, downregulated (HSP90) in AD group. Altered expression of all these up- or downregulated proteins may be linked to AD pathophysiology. Thus, our results are encouraging for searching new biomarker candidates for AD. TF is potential biomarker candidate, because TF seems to reflect altered protein levels already in mild AD dementia.

Unraveling the proteomic signatures of coronary artery disease and hypercholesterolemia.

Atherosclerosis, a leading cause of coronary artery disease (CAD), is heavily influenced by hypercholesterolemia (HC). Proteomics research has shown promise in identifying biological markers for CAD diagnosis and prognosis. This cross-sectional study aimed to identify novel biomarkers for detecting HC and CAD. Through the analysis of proteome data from healthy controls (n=45) and patients diagnosed with HC (n=51) or CAD (n=32), distinct protein patterns associated with each condition were identified. Significant alterations in protein levels were identified with a false discovery rate (FDR)-corrected q-value of <0.05. Subsequent receiver operating characteristic (ROC) analysis, with an area under the curve (AUC) greater than 0.75, was conducted. CAD patients exhibited significantly increased levels of the cholesterol-metabolizing protein PCSK9 and varied levels of the angiogenesis-related protein SDF-1 compared to controls. In pairwise comparisons among the study groups, 65 proteins showed significant differential expression. Notably, 14 of these proteins had significant correlations with blood cholesterol levels. Additionally, 22 of the identified proteins were associated with CAD or HC pathways, with nine proteins being common to both conditions (Apo E, Apo E3, MMP-3, PCSK9, SDF-1, Apo B, PAFAH, HSP 60, and TAK1-TAB1). Nevertheless, this is an exploratory study, and validation studies are needed to confirm these potential protein biomarkers for CAD in the context of HC.

The effects of chronic desipramine treatment on neurotrophin-3 in the brain of mice with selective depletion of CREB and CREM in noradrenergic neurons

The disturbances in neurotrophic support are thought to be one of the main causes of depression, which depend not only on the neurotrophins themselves but also on the molecules regulating their synthesis and effector functions. One such molecule is cAMP responsive element binding protein (CREB), which role in depression and antidepressant drugs mechanism of action has been extensively studied. However, CREB’s effects vary depending on brain structure, necessitating specific transgenic models for studying its function. Moreover, deletion of CREB enhances cAMP response element modulator (CREM) expression, suspected to compensate for CREB in its absence.Previously, mice lacking CREB in noradrenergic neurons and CREM (Creb1DbhCreCrem-/-) showed to be insensitive to acute desipramine, whereas mice lacking only CREB (Creb1DbhCre) showed similar effects as wild type animals (w/t). As neurotrophic changes require chronic antidepressant treatment, in current study mice (w/t, Creb1DbhCre and Creb1DbhCreCrem-/-; both males and females) were given desipramine for 21 days, to assess the effects of the drug on CREB, neurotrophins and their receptors in the hippocampus and prefrontal cortex.Interestingly, desipramine had no effect on CREB in neither of studied groups. However, both male and female mice lacking CREB and CREM displayed alterations in neurotrophin-3 (NTF3) expression or protein levels, modulated by desipramine. These findings suggest NTF3 is connected with inhibited response to acute and probably chronic desipramine administration in Creb1DbhCreCrem–/– mice, although in w/t chronic desipramine had no effect on NTF3. Nevertheless, our findings give insight into the role of non-BDNF neurotrophins in the mechanism of antidepressant drugs.

GHRH-stimulated pituitary small extracellular vesicles inhibit hepatocyte proliferation and IGF-1 expression by its cargo miR-375-3p

Small extracellular vesicles (sEV) have emerged as a novel mode of intercellular material transport and information transmission. It has been suggested hormones may regulate the production and function of sEV. However, the specific impact of growth hormone-releasing hormone (GHRH) on pituitary sEV production and the role of sEV in the regulation of the GHRH-GH-IGF axis has not been previously reported. The results of the present study demonstrated that GHRH increased the production of pituitary sEV by promoting the expression of Rab27a. More importantly, GHRH induced alterations in protein and miRNA levels within GH3-sEV components. Notably, GH3-sEV with GHRH treatment exhibited the enhanced ability to impede BRL 3A cell proliferation and the expression of IGF-1. Conclusively, for the first time, we corroborate the influence of GHRH on pituitary sEV, thereby presenting novel evidence for how sEV participates in the balance of the GHRH-GH-IGF axis. Importantly, this study provides new insight into a novel balance mechanism mediated by sEV within the endocrine system.Graphical

Indigo alleviates psoriasis through the AhR/NF-κB signaling pathway: an in vitro and in vivo study.

Psoriasis is a chronic inflammatory skin disease. A strong association between the AhR/ NFκB axis and the inflammatory response in psoriasis. Indigo (IDG) has demonstrated significant anti-inflammatory properties. This study aimed to assess the anti-psoriatic efficacy of IDG while investigating the underlying mechanisms involved. In the in vitro experiments, cell viability was assessed using the CCK-8. qRT-PCR was employed to measure the mRNA levels of NF-κB, TNF-α, IL-1β, AhR, and CYP1A1. Western blotting was conducted to examine alterations in cytoplasmic and nuclear AhR protein levels. Additionally, an IDG nanoemulsion (NE) cream was prepared for the in vivo experiments. A psoriasis-like skin lesion mice model was induced using IMQ (62.5 mg/day for 7 days). The severity of psoriasis was evaluated using PASI, and skin lesions were scored while epidermal thickness was assessed via HE staining. The expression of inflammatory markers, including IL-6, IL-13, IL-17A, MCP-1, and TNF-α, was detected in skin lesions using Luminex. The levels of CYP1A1, p65, and p-p65 proteins were determined by Western blotting. LPS stimulation significantly elevated TNF-α, IL-6, and NF-κB mRNA levels, which were notably reduced by IDG treatment. Additionally, IDG significantly enhanced the expression of AhR and CYP1A1 mRNA. Further investigation revealed that IDG facilitated AhR translocation from the cytoplasm to the nucleus. In the IMQ-induced psoriasis-like mouse model, IDG NE substantially ameliorated the severity of skin lesions. Moreover, IDG NE treatment reduced the upregulation of inflammatory cytokines such as IL-6, IL-17A, MCP-1, and TNF-α in IMQ-induced skin lesions. It was also observed that IDG NE treatment increased CYP1A1 protein expression while inhibiting p65 and p-p65 protein expression. IDG emerges as a promising treatment for psoriasis, demonstrating effective therapeutic outcomes. Its mechanism of action is likely linked to the modulation of the AhR/NFκB signaling pathway.

Identification of Potential Drug Targets for Sleep Apnea Through Mendelian Randomization

Background: Sleep apnea is a significant health impediment, and it presently lacks efficacious therapeutic interventions. Thus, it is imperative to discover novel therapeutic targets that could guide clinical treatment strategies. Objective: This study aims to utilize an integrative analytic approach to unearth previously unappreciated protein-encoding genes implicated in sleep apnea susceptibility. Methods: Through the Multi-Marker Analysis of Genomic Annotation (MAGMA), we aligned Single-Nucleotide Polymorphism (SNP) summary statistics from Genome-Wide Association Studies (GWAS) of gene bodies to discern potential risk genes. Following the MAGMA results, we conducted a round of Transcriptome-Wide Association Studies (TWAS) and Proteome-Wide Association Studies (PWAS) to expedite the conversion of genetic associations into probable protein targets. Mendelian Randomization (MR) and co-localization analysis were employed to ascertain the causal linkage between the candidate target genes and sleep apnea. Finally, a mediation analysis was undertaken to explore the possible intermediary role of 150 inflammatory metabolites and 1,124 proteins. Results: The MAGNA analysis revealed 2,819 genes in association with sleep apnea. TWAS and PWAS analyses indicated that cis-regulation of nine particular genes could play a role in sleep apnea onset via blood protein level alterations. MR and co-localization analyses also suggested a causal relationship with sleep apnea for three genes (ACADVL, CCDC134, UPP1). Consistent associations were found between genetically predicted biomarkers and Albumin, HCC-1, N-acetyl carnosine, and RELT, pointing towards their potential mediating roles in sleep apnea's etiological pathway. result: The MAGNA analysis revealed 2,819 genes in association with sleep apnea. TWAS and PWAS analyses indicated that cis-regulation of nine particular genes could play a role in sleep apnea onset via blood protein level alterations. MR and co-localization analyses also suggested a causal relationship with sleep apnea for three genes (ACADVL, CCDC134, UPP1). Consistent associations were found between genetically predicted biomarkers and Albumin, HCC-1, N-acetylcarnosine, and RELT, pointing towards their potential mediating roles in sleep apnea&amp;#039;s etiological pathway. Conclusion: Our findings indicate that ACADVL, CCDC134, and UPP1 genes are potentially significant targets for further functional investigation and therapeutic interventions for sleep apnea.

Orbital Blood Flow and Pseudoexfoliation Syndrome - A Narrative Review

Abstract The narrative review titled ‘Orbital Blood Flow and Pseudoexfoliation Syndrome’ provides a comprehensive exploration of the intricate interplay between pseudoexfoliation (PEX) syndrome and orbital blood flow, offering insights into its clinical ramifications. Commencing with an overview of the historical context and prevalence of PEX, the review traverses various dimensions, including gender and age considerations, mortality/morbidity rates, and the underlying pathophysiology. It delves into the composition of PEX material and delineates the differential gene expression associated with the syndrome. Central to the discourse is the impact of PEX on orbital vasculature, elucidating its correlation with ocular ischemia and alterations in protein levels within the aqueous humour. Techniques for clinical evaluation, notably colour Doppler imaging, emerge as indispensable tools for assessing blood flow in patients with PEX.The article emphasises the significance of vascular dysfunction in PEX, highlighting the findings of decreased blood flow velocities and elevated resistive indices in affected individuals.The review underscores the imperative of comprehending the vascular implications of PEX to optimise diagnostic and management approaches, with far reaching implications for minimising operative complications and enhancing patient outcomes.

Subacute Toxicity and Pharmacokinetic Evaluation of the Synthetic Cannabinoid 4F-MDMB-BUTINACA in Rats: A Forensic and Toxicological Perspective

Background: 4-MDMB-BUTINACA, a next-generation synthetic cannabinoid, presents significant public health and forensic challenges due to its evolving nature and potential toxicity. Methods: This study evaluates the subacute toxic effects and pharmacokinetics of 4−Fluoro MDMB−BUTINACA (4F-MDMB-BUTINACA) in adult male albino rats, administered orally for seven days at doses of 1 mg/kg, 5 mg/kg, and 15 mg/kg. The hematological, biochemical, and histopathological parameters were assessed and compared to controls. Results: The pharmacokinetics were determined using GC–MS/MS with a positive chemical ionization and granisetron as an internal standard. A histological analysis revealed inflammatory cell aggregation, congestion, hemorrhage, edema, and fibrosis in various tissues, with renal examinations showing tubule degradation, glomerular atrophy, Bowman’s space expansion, edema, and hemorrhage. The liver exhibited cellular infiltration, while cardiac muscle fibers showed myocardial fiber degradation and inflammatory cell aggregation. Biochemical assays indicated significant alterations (p &lt; 0.05) in the serum levels of AST, ALT, ALP, GGT, total protein, albumin, triglycerides, urea, MCHC, MCV, RDW, platelets, neutrophils, eosinophils, and basophils compared to the controls. Conclusions: The validated bioanalytical method revealed rapid absorption of 4F-MDMB-BUTINACA, with a plasma half-life of 2.371 h, a volume of distribution of 2272.85 L, and a plasma clearance rate of 664.241 L/h. In conclusion, 4F-MDMB-BUTINACA is a highly toxic synthetic cannabinoid, particularly affecting the liver, kidneys, and heart.

Multiomics profiling uncovers curdione-induced reproductive toxicity in HTR-8/SVneo cells

The assessment of medication toxicity and safety is pivotal during pregnancy. Curdione, a sesquiterpene compound extracted from Curcumae Radix, displays beneficial properties in terms of anti-inflammatory, tumor growth suppression, and anti-coagulative effects. However, its reproductive toxicity and precise mechanism remain unclear. This study aims to explore the mechanism of curdione-induced toxicity damage in HTR-8/SVneo cells through the epigenetics, proteomics, and metabolomics, and experimental verification. The results showed that curdione elicited alterations in m6A modification, gene expression, protein levels, and cellular metabolism of HTR-8/SVneo cells. Additionally, curdione induces oxidative stress, mitochondrial and DNA damage, while also downregulating the expression of Wnt6, β-catenin, ZO-1, and CLDN1 proteins. Curdione has the potential to modulate oxidative stress, mitochondrial dysfunction, and disruption of tight junctions via the Wnt/β-catenin signaling pathway, which contributes to cellular damage in HTR-8/SVneo cells.

High glucose induces DNA methyltransferase 1 dependent epigenetic reprogramming of the endothelial exosome proteome in type 2 diabetes

In response to hyperglycemia, endothelial cells (ECs) release exosomes with altered protein content and contribute to paracrine signalling, subsequently leading to vascular dysfunction in type 2 diabetes (T2D). High glucose reprograms DNA methylation patterns in various cell/tissue types, including ECs, resulting in pathologically relevant changes in cellular and extracellular proteome. However, DNA methylation-based proteome reprogramming in endothelial exosomes and associated pathological implications in T2D are not known. Hence, in the present study, we used Human umbilical vein endothelial cells (HUVECs), High Fat Diet (HFD) induced diabetic mice (C57BL/6) and clinical models to understand epigenetic basis of exosome proteome regulation in T2D pathogenesis . Exosomes were isolated by size exclusion chromatography and subjected to tandem mass tag (TMT) labelled quantitative proteomics and bioinformatics analysis. Immunoblotting was performed to validate exosome protein signature in clinically characterized individuals with T2D. We observed ECs cultured in high glucose and aortic ECs from HFD mouse expressed elevated DNA methyltransferase1 (DNMT1) levels. Quantitative proteomics of exosomes isolated from ECs treated with high glucose and overexpressing DNMT1 showed significant alterations in both protein levels and post translational modifications which were aligned to T2D associated vascular functions. Based on ontology and gene-function-disease interaction analysis, differentially expressed exosome proteins such as Thrombospondin1, Pentraxin3 and Cystatin C related to vascular complications were significantly increased in HUVECs treated with high glucose and HFD animals and T2D individuals with higher levels of glycated hemoglobin. These proteins were reduced upon treatment with 5-Aza-2’-deoxycytidine. Our study shows epigenetic regulation of exosome proteome in T2D associated vascular complications.

Targeting NLRP3 inhibits AML progression by inducing PERK/eIF2-mediated apoptosis

BackgroundAcute myeloid leukemia (AML) is characterized by the abnormal proliferation of myeloid precursor cells and presents significant challenges in treatment due to its heterogeneity. Recently, the NLRP3 inflammasome has emerged as a potential contributor to AML pathogenesis, although its precise mechanisms remain poorly understood.MethodsPublic genome datasets were utilized to evaluate the expression of NLRP3 inflammasome-related genes (IL-1β, IL-18, ASC, and NLRP3) in AML patients compared to healthy individuals. CRISPR/Cas9 technology was employed to generate NLRP3-deficient MOLM-13 AML cells, followed by comprehensive characterization using real-time PCR, western blotting, FACS analysis, and transmission electron and immunofluorescence microscopy. Proteomic analyses were conducted to identify NLRP3-dependent alterations in protein levels, with a focus on the eIF2 kinase PERK-mediated signaling pathways. Additionally, in vivo studies were performed using a leukemic mouse model to elucidate the pathogenic role of NLRP3 in AML.ResultsElevated expression of NLRP3 was significantly associated with diminished overall survival in AML patients. Genetic deletion, pharmacological inhibition and silencing by RNA interference of NLRP3 led to decreased AML cell survival through the induction of apoptosis. Proteomic analyses uncovered NLRP3-dependent alterations in protein translation, characterized by enhanced eIF2α phosphorylation in NLRP3-deficient AML cells. Moreover, inhibition of PERK-mediated eIF2α phosphorylation reduced apoptosis by downregulating pro-apoptotic Bcl-2 family members. In vivo studies demonstrated reduced leukemic burden in mice engrafted with NLRP3 knockout AML cells, as evidenced by alleviated leukemic symptoms.ConclusionOur findings elucidate the involvement of the NLRP3/PERK/eIF2 axis as a novel driver of AML cell survival. Targeting NLRP3-induced signaling pathways, particularly through the PERK/eIF2 axis, presents a promising therapeutic strategy for AML intervention. These insights into the role of the NLRP3 inflammasome offer potential avenues for improving the prognosis and treatment outcomes of AML patients.

The effects of Lactobacillus plantarum PS128 in patients with major depressive disorder: an eight-week double-blind, placebo-controlled study

Major depressive disorder (MDD) is a complex mental disorder, potentially linked to the gut-microbiota-brain axis. Probiotics like Lactobacillus plantarum PS128 (PS128) may improve depressive symptoms by modulating the gut microbiota based on our previous open trial. We conducted an 8-week double-blind, placebo-controlled trial to investigate the impact of PS128 on depression severity, markers of inflammation and gut permeability, and the gut microbiota composition in 32 patients with MDD with stable antidepressant treatment but moderate symptom severity. Following the 8-week intervention, both the Hamilton Depression Rating Scale-17 score (HAMD), and Depression and Somatic Symptoms Scale (DSSS) showed a significant decrease in both groups (p<0.001). However, there was no significant difference in the change of depression severity between groups (p=0.203). Moreover, alterations in serum levels of high sensitivity C-reactive protein, interleukin-6, tumor necrosis factor-α, and intestinal fatty acid binding protein, as well as changes in the gut microbiota composition, did not exhibit significant differences before and after intervention or between the groups. In comparison to the placebo group, our study did not find significant effects of PS128 on depressive symptoms, biomarkers of inflammation and gut permeability, and the overall gut microbiota composition. Nonetheless, we observed a potential impact of PS128 on the symbiosis of specific taxa. To comprehensively understand the psychophysiological effects of PS128 in patients with MDD, further research with a larger sample size is imperative.