Identification and targeting of metastatic biomarkers for hepatocellular carcinoma therapeutics using small molecules library of curcumin analogues.

The understanding of the molecular basis of complex diseases like hepatocellular carcinoma (HCC) needs large datasets of multiple genes and proteins involved in different phenomenon of its development. This study focuses on the molecular basis of HCC and the development of therapeutic strategies. We analyzed a dataset of 5475 genes (Homo sapiens) involved in HCC hallmarks, involving comprehensive data on multiple genes and frequently mutated genes. As HCC is characterized by metastasis, angiogenesis, and oxidative stress, exploration of genes associated with them has been targeted. Through gene ontology, functional characterization, and pathway enrichment analysis, we identified target proteins such as Lysyl oxidase, Survivin, Cofilin, and Cathepsin B. A library of curcumin analogs was used to target these proteins. Tetrahrydrocurcumin showed promising binding affinities for all four proteins, suggesting its potential as an inhibitor against these proteins for HCC therapy.

The cross-talk between mucormycosis, steroids and diabetes mellitus amidst the global contagion of COVID-19.

Mucormycosis is an opportunistic fungal disease that targets individuals having an impaired immune system due to a wide array of risk factors including HIV-AIDS, immunosuppressive therapy, diabetes mellitus, etc. The current explosive outbreak of coronavirus disease 2019 (COVID-19) has become the latest threat to such patients who are already susceptible to secondary infections. Physiological outcomes of COVID-19 end up in a cascade of grave alterations to the immunological profile and irreparable harm to their respiratory passage, heart and kidneys. Corticosteroidal treatment facilitates faster recovery and alleviates the adverse pathological effects of the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). But clinical reports lend this approach a darker perspective especially if these patients have pre-existing diabetes mellitus. The mucormycotic fungal genera belonging to the order Mucorales not only survive but thrive under the comorbidity of COVID-19 and diabetes, often staying undetected until they have inflicted irreversible damage. Steroidal usage has been noted to be a common thread in the sudden spurt in secondary fungal infections among COVID-19 cases. Once considered a rare occurrence, mucormycosis has now acquired a notoriously lethal status in mainstream medical hierarchy. We set out to investigate whether corticosteroidal therapy against COVID-19 emboldens the development of mucormycosis. We also assess the conditions brought forth by steroidal usage and uncontrolled progression of diabetes in COVID-19 cases and their effect on the susceptibility towards mucormycosis.

Hydrogenated cove-edge aluminum nitride nanoribbons for ultrascaled resonant tunneling diode applications: a computational DFT study.

While synthesizing quasi-one-dimensional nanoribbons, there is a finite probability that edges have cove-edge defects. This paper focuses on the structural, electronic, and transport properties of cove-edge aluminum nitride nanoribbons (AlNNR) using density functional theory and the non-equilibrium Green's function (NEGF) method. The cove-edge AlNNRs are thermodynamically stable and exhibit metallic behavior. Interestingly, the calculated current-voltage characteristics of the cove-edge AlNNR-based nanodevices show negative differential resistance (NDR). The H-AlN-Cove nanodevice exhibits high peak-to-valley current ratio (PVCR) of the order of 107. The calculated PVCR of the H-AlN-Cove nanodevice is 106times higher than that of the silicene nanoribbon (SiNR) and graphene nanoribbon (GNR), and 104times higher than that of the phosphorene nanoribbon (PNR) and arsenene nanoribbons (ANR)-based devices respectively. The NDR feature with high PVCR provides a prospect for the cove-edge AlNNR in nanodevice applications.

Nitrogen-doped zinc oxide nanoribbons for potential resonant tunneling diode applications.

Armchair ZnONRs doped with nitrogen are investigated in the current manuscript for possible applications based on negative differential resistance (NDR). To conduct the theoretical research, we use density functional theory (DFT) in conjunction with the non-equilibrium Green's function (NEGF) formalism to carry out first principles computations. Pristine ZnONR (P-ZnONRs) is a semiconductor with a wide energy bandgap (Eg) of 2.53 eV. However, one edge N-doped ZnONRs (SN-ZnO) and both edge N-doped ZnONRs (DN-ZnO) are metallic. Partial density of states (PDOS) reveals that the metallicity is caused by the doped nitrogen atom. The transport characteristics analysis revealed the negative differential resistance (NDR) characteristics in the N-doped ZnONRs. The peak-to-valley current ratios (PVCR) are computed and measured to be 4.58 × 1021 and 1.83 × 1022 for SN-ZnO and DN-ZnO, respectively. The obtained findings suggest the significant potential of armchair ZnONRs for NDR-based applications such as switches, rectifiers, oscillators, memory devices, etc.

Nanointerconnect design based on edge fluorinated/hydrogenated zigzag borophene nanoribbons: an ab initio analysis.

Using an ab initio framework and non-equilibrium Green's function technique, the effect of hydrogen and fluorine atom passivation on the electronic and transport properties of borophene nanoribbons (BNRs) are explored. For zigzag edge states, we have explored all potentially stable combinations of hydrogen and fluorine passivation. Fluorine passivation leads to thermodynamically stable structures with improved stability for the increased concentration of F atoms, according to our binding energy (Eb) calculations. Furthermore, density-of-states and dispersion relation (E-k structures) computations indicate that fluorine-passivated BNRs are primarily metallic in nature. We proposed these nanostructures for their use in metal interconnects because of their increased metallicity. We have used the typical two-probe setup to calculate the critical parameters like quantum resistance (RQ), kinetic inductance (LK), and quantum capacitance (CQ) to evaluate their performance as metal interconnects. Because they have the lowest estimated values of LK = 26.1 nH µm-1, and CQ = 399 pF cm-1, the zigzag BNRs (ZBNRs) with two edge fluorinated (F-BNR-F) nanostructures may be considered as a promising candidate for nanoscale interconnect applications.

MicroRNA expression in extracellular vesicles as a novel blood-based biomarker for Alzheimer's disease.

INTRODUCTION: Brain cell-derived small extracellular vesicles (sEVs) in blood offer unique cellular and molecular information related to the onset and progression of Alzheimer's disease (AD). We simultaneously enriched six specific sEV subtypes from the plasma and analyzed a selected panel of microRNAs (miRNAs) in older adults with/without cognitive impairment. METHODS: Total sEVs were isolated from the plasma of participants with normal cognition (CN; n = 11), mild cognitive impairment (MCI; n = 11), MCI conversion to AD dementia (MCI-AD; n = 6), and AD dementia (n = 11). Various brain cell-derived sEVs (from neurons, astrocytes, microglia, oligodendrocytes, pericytes, and endothelial cells) were enriched and analyzed for specific miRNAs. RESULTS: miRNAs in sEV subtypes differentially expressed in MCI, MCI-AD, and AD dementia compared to the CN group clearly distinguished dementia status, with an area under the curve (AUC) > 0.90 and correlated with the temporal cortical region thickness on magnetic resonance imaging (MRI). DISCUSSION: miRNA analyses in specific sEVs could serve as a novel blood-based molecular biomarker for AD. HIGHLIGHTS: Multiple brain cell-derived small extracellular vesicles (sEVs) could be isolated simultaneously from blood. MicroRNA (miRNA) expression in sEVs could detect Alzheimer's disease (AD) with high specificity and sensitivity. miRNA expression in sEVs correlated with cortical region thickness on magnetic resonance imaging (MRI). Altered expression of miRNAs in sEVCD31 and sEVPDGFRß suggested vascular dysfunction. miRNA expression in sEVs could predict the activation state of specific brain cell types.

Liquid wastes as a renewable feedstock for yeast biodiesel production: Opportunities and challenges.

Microbial lipids (bacterial, yeast, or algal) production and its utilization as a feedstock for biodiesel production in a sustainable and economical way along with waste degradation is a promising technology. Oleaginous yeasts have demonstrated multiple advantages over algae and bacteria such as high lipid yields, lipid similarity to vegetable oil, and requirement of lesser area for cultivation. Oleaginous yeasts grown on lignocellulosic solid waste as renewable feedstocks have been widely reported and reviewed. Recently, industrial effluents and other liquid wastes have been evaluated as feedstocks for biodiesel production from oleaginous yeasts. The idea of the utilization of wastewater for the growth of oleaginous yeasts for simultaneous wastewater treatment and lipid production is gaining attention among researchers. However, the detailed knowledge on the economic aspects of different process involved during the conversion of oleaginous yeast into lipids hinders its large-scale application. Therefore, this review aims to provide an overview of yeast-derived biodiesel production by utilizing industrial effluents and other liquid wastes as feedstocks. Various technologies for biomass harvesting, lipid extraction and the economic aspects specifically focused on yeast biodiesel production were also analyzed and reported in this review. The utilization of liquid wastes and the incorporation of cost-efficient harvesting and lipid extraction strategy would facilitate large-scale commercialization of biodiesel production from oleaginous yeasts in near future.

Network pharmacological evaluation of strigolactones efficacy as potential inhibitors against therapeutic targets of hepatocellular carcinoma.

PURPOSE: Hepatocellular carcinoma (HCC) is the uncontrolled growth of hepatocytes which results in nearly 5 million deaths worldwide. Specific strategies have been developed to treat HCC, including surgery, chemotherapy and radiotherapy. But, the effective disease dealing requires synergistic collaboration with other approaches, which often results in moderate to severe side effects during and after the treatment period. Therefore, the focus is now shifting to explore and retrieve those plant-based products that could be utilized to treat HCC with maximum efficacy without causing any side effects. Strigolactones (SL) are compounds of plant origin derived from Striga lutea responsible for controlling the branching pattern of stem and have reported anti-cancerous activity by promoting apoptosis at micromolar concentrations. However, little work has been done concerning determining the pharmacogenomic effect of strigolactones on HCC. METHODS: Current work focuses on comparing therapeutic efficiencies of SL analogs against core targets of HCC using network pharmacology approach, pharmacokinetics analysis, gene ontogeny, functional enrichment analysis, molecular docking and Molecular Dynamics simulation. RESULTS: Drug-target prediction and functional enrichment analysis showed that HDAC1 and HDAC2 are the core proteins involved in hepatocellular carcinoma that strigolactone analogs can target. Consequently, results from molecular docking and MD simulation analyses report that among all the SL analogs strigol, epistrigol and nijmegen1 can turn out to be most effective in downregulating the expression of HDAC1, HDAC2 and CYP19A. CONCLUSION: Strigol, epistrigol and nijmegen1 could be used as potential inhibitors against HCC and can be further validated through in vitro/in vivo studies.

Evaluation of the effects of 0.05% sodium hypochlorite and 0.12% chlorhexidine gluconate twice daily rinse on periodontal parameters and gingival crevicular fluid HSV1 and CMV levels in patients with chronic periodontitis: a multicentric study.

Background: Mechanical debridement of periodontal pockets remains the mainstay of therapy in all forms of periodontitis. There is 47% greater reduction in plaque amount when sodium hypochlorite (NaOCl) is used as an adjunct when compared with water rinsing. The aim of this study was to evaluate the effects of 0.05% NaOCl and 0.12% chlorhexidine gluconate twice daily rinse on periodontal parameters and gingival crevicular fluid (GCF) HSV1 and CMV levels in chronic periodontitis. Methods: Patients assigned to group A were prescribed 0.05% NaOCl mouthwash for twice daily rinse. Patients in group B were prescribed 0.12% chlorhexidine gluconate mouthwash to be used twice daily. Evaluation of periodontal parameters was done at baseline and after six months following therapy. GCF HSV1 and CMV levels were evaluated using a polymerase chain reaction. Results: A statistically significant difference was noted in the improvement in periodontal parameters between both groups, when evaluated six months following therapy with greater reduction in group A vis-a-vis group B. Conclusion: NaOCl when prescribed as a twice daily mouthwash can be recommended as a part of the home care regime in patients with chronic periodontitis. It is more cost-effective, easily available and can be beneficial to the troops in difficult terrains and extremes of climates, where oral healthcare facilities are not easily accessible.

Alteration of proteome in germinating seedlings of piegonpea (Cajanus cajan) after salt stress.

Pigeonpea (Cajanus cajan) is an important crop in semi-arid regions and a significant source of dietary proteins in India. The plant is sensitive to salinity stress, which adversely affects its productivity. Based on the dosage-dependent influence of salinity stress on the growth and ion contents in the young seedlings of pigeonpea, a comparative proteome analysis of control and salt stressed (150 mM NaCl) plants was conducted using 7 days-old seedlings. Among various amino acids, serine, aspartate and asparagine were the amino acids that showed increment in the root, whereas serine, aspartate and phenylalanine showed an upward trend in shoots under salt stress. Furthermore, a label-free and gel-free comparative Q-Tof, Liquid Chromatography-Mass spectrometry (LC-MS) revealed total of 118 differentially abundant proteins in roots and shoots with and without salt stress conditions. Proteins related to DNA-binding with one finger (Dof) transcription factor family and glycine betaine (GB) biosynthesis were differentially expressed in the shoot and root of the salinity-stressed seedlings. Exogenous application of choline on GB accumulation under salt stress showed the increase of GB pathway in C. cajan. Gene expression analysis for differentially abundant proteins revealed the higher induction of ethanolamine kinase (CcEthKin), choline-phosphate cytidylyltransferase 1-like (CcChoPh), serine hydroxymethyltransferase (CcSHMT) and Dof protein (CcDof29). The results indicate the importance of, choline precursor, serine biosynthetic pathways and glycine betaine synthesis in salinity stress tolerance. The glycine betaine protects plant from cellular damages and acts as osmoticum under stress condition. Protein interaction network (PIN) analysis demonstrated that 61% of the differentially expressed proteins exhibited positive interactions and 10% of them formed the center of the PIN. Further, The PIN analysis also highlighted the potential roles of the cytochrome c oxidases in sensing and signaling cascades governing salinity stress responses in pigeonpea. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s12298-021-01116-w.

Diabetic kidney disease: A systematic review on the role of epigenetics as diagnostic and prognostic marker.

Diabetic kidney disease is one of the most serious microvascular complications and among the leading causes of end stage renal disease. Persistently increasing albuminuria has been considered to be the central hallmark of nephropathy. However, albuminuria can indicate kidney damage for clinicians; it is not a specific biomarker for prediction of diabetic kidney disease prior to the onset of this devastating complication, and in fact all individuals with microalbuminuria do not progress to overt nephropathy. Controlled glycemia is unable to prevent nephropathy in all diabetic individuals indicating the role of other factors in progression of diabetic kidney disease. There are numerous cellular and molecular defects persisting prior to appearance of clinical symptoms. So, there is an urgent need to look for easy, novel, and accurate way to detect diabetic kidney disease prior to its beginning or at the infancy stage so that its progression can be slowed or arrested. It is now accepted that initiation and progression of diabetic kidney disease are a result of complex interactions between genetic and environmental factors. Environmental signals can alter the intracellular pathways by chromatin modifiers and regulate gene expression patterns leading to diabetes and its complications. In the present review, we have discussed a possible link between aberrant DNA methylation and altered gene expression in diabetic kidney disease. Drugs targeting to reverse epigenetic alteration can retard or stop the development of this devastating disease, just by breaking the chain of events occurring prior to the development of this microvascular complication in patients with diabetes.

Controlling pathogenesis in Candida albicans by targeting Efg1 and Glyoxylate pathway through naturally occurring polyphenols.

Candida albicans has frequently shown resistance to azoles, the commonly used antifungal drugs. Efg1 has dual role under normoxia and hypoxia supporting infection. It is the major regulator of morphogenesis in C. albicans requisite for its pathogenesis. Targeting this protein is expected to render Candida ineffective to undergo filamentation causing virulence. Further the glyoxylate pathway supports the stress resistance and pathogenesis. In the present study an in silico approach and in vitro validation has been performed to find the potential role of polyphenols in controlling hyphal growth in C. albicans. The aspect of changes biome which may provide required niche to the pathogen has been checked which certainly opens the doors towards safe natural polyphenol-based drugs as potent antifungals.

Prevention of OprD regulated antibiotic resistance in Pseudomonas aeruginosa biofilm.

In P.aeruginosa biofilms, the issue of antibiotic resistance is of particular importance due to increasing number of infections being reported in medical implants. The current study is focused on CzcR and CopR proteins which are part of two-component signal transduction systems (TCSs) - CzcR-CzcS and CopR-CopS respectively in P.aeruginosa. They both negatively regulate OprD porin expression which affects the intake of antibiotics like carbapenems. These two proteins can be treated as targets to combat antibiotic resistance in P.aeruginosa. Docking was performed on these proteins in search of inhibitors against the CzcR-CzcS and CopR-CopS TCSs. Efficient inhibitory ligands were evaluated on the basis of least binding energy, human oral absorption and ADME properties using a four-tier structure based virtual screening. The resulting ligands displayed high effective inhibitory property and satisfactory pharmacokinetics as compared to inhibitors which have been identified before for two-component signal transduction systems for gram negative bacteria. These potential inhibitors can now be used further in wet lab by performing selectivity assays to determine their inhibition rate against P.aeruginosa biofilms. Identification of potential leads may enable the development of new therapeutic strategies aimed at disrupting P.aeruginosabiofilms.

Prevention of IcaA regulated poly N-acetyl glucosamine formation in Staphylococcus aureus biofilm through new-drug like inhibitors: In silico approach and MD simulation study.

OBJECTIVES: The effectiveness of various ligands against the protein structure of IcaA of the IcaABCD gene locus of Staphylococcus aureus were examined using the approach of structure based drug designing in reference with the protein's efficiency to form biofilms. RESULTS: Four compounds CID42738592, CID90468752, CID24277882, and CID6435208 were secluded from a database of 31,242 inhibitory ligands on the justification of the evaluated values falling under the four - tier structure based virtual screening. Under this principle value of least binding energy, human oral absorption and ADME properties were taken into consideration. Using the Glide module of Schrödinger, the above mentioned ligands showed an effective action against the protein IcaA which showed reduced activity as a glucosaminyl transferase. The complex of protein and ligand with best docking score was chosen for simulation studies. CONCLUSIONS: Structure based drug designing for the protein IcaA has given us potential leads as anti - biofilm agents. These screened out ligands might enable the development of new therapeutic strategies aimed at disrupting Staphylococcus aureus biofilms. The complex was showing stability towards the end of time for which it has been put for simulation. Thus molecule could be considered for making of biofilms.

Restraining Pathogenicity in Candida albicans by Taxifolin as an Inhibitor of Ras1-pka Pathway.

Candida albicans is one of the most virulent and opportunistic fungal strains. In the present scenario, majority metabolic imbalances and unsuccessful treatments of some severe diseases including cancer, diabetes, HIV, psoriasis are because of invasive Candida emergence. Being a beneficial integral part of human biome, its elimination is not possible. The major pathogenicity characteristics in Candida involve hyphal growth, biofilm formation, HSP90 down regulation and genetic modifications. Ras1-pka pathway initiated by HSP90 down regulation is important for hyphal growth and has been focused in the present study. The principle transcriptional factors that induce hyphal growth causing invasiveness and virulence through this pathway have been identified as Tec1 and Rfg1. In the present study, taxifolin, a naturally occurring polyphenol, has been identified as inhibitor for both the transcriptional factors in parallel.

The Association Between Oxidative Stress and the Progression of Heart Failure: A Systematic Review.

Chronic heart failure (CHF) is a progressive multifactorial condition where the role of oxidative stress may have implications in the pathogenesis of the disease. Despite growing interest among researchers and clinicians, the limited, unorganized, and divergent findings regarding the association between oxidative stress and the progression of heart failure (HF) have prompted us to conduct this study. Drawing upon the evolving nature of this research domain, this study is one of the first of its kind to present a systematic and comprehensive overview of the existing evidence regarding the role of oxidative stress production in the progression of HF. This study systematically reviews peer-reviewed empirical studies published in English, particularly focusing on the association between oxidative stress and the progression of HF. Parameters, such as publication year, study design, population demographics (size, age, and gender), types of HF, and characterization of markers in the existing studies, were reviewed. Following the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) procedure, a thorough search was conducted on PubMed, Cochrane, Embase, and Sage databases, without any restrictions on the publication dates of articles, which yielded a total of 1,808 records on the association of oxidative stress production with clinical outcomes in HF patients. The analysis of the content of 17 articles offered a robust observation of this phenomenon, providing insights into the levels of oxidative stress, antioxidant markers, and the enzymes involved in the production of reactive oxygen species (ROS), and their association with the progression and severity of HF. The findings highlighted various knowledge gaps and future research priorities are recommended in the areas of interest and unexplored areas.

Comparative evaluation of divergent concoction of NGF, BDNF, EGF, and FGF growth factor's role in enhancing neuronal differentiation of adipose-derived mesenchymal stem cells.

MSCs (Mesenchymal Stem Cells) can differentiate into various lineages, including neurons and glial cells. In the past few decades, MSCs have been well explored in the context of neuronal differentiation and have been reported to have the immense potential to form distinct kinds of neurons. The distinguishing features of MSCs make them among the most desired cell sources for stem cell therapy. This study involved the trans-differentiation of Adipose-derived human Mesenchymal Stem Cells (ADMSCs) into neurons. The protocol employs a cocktail of chemical inducers in different combinations, including Brain-derived neurotrophic factor (BDNF), epidermal growth factor (EGF), and Nerve growth factor (NGF) Fibroblastic growth factor (FGF), in induction media. Both types have been successfully differentiated into neurons, confirmed by morphological aspects and the presence of neural-specific markers through RT-PCR (Reverse transcription polymerase chain reaction) studies and immunocytochemistry assay. They have shown excellent morphology with long neurites, synaptic connections, and essential neural markers to validate their identity. The results may significantly contribute to cell replacement therapy for neurological disorders.

A Retrospective Observational Study of the Microbial Etiology and Antimicrobial Susceptibility Patterns of Blood Cultures From ICU Patients at a Healthcare Facility in North India.

Introduction Bloodstream infections (BSI) are a leading source of fatalities and morbidity in hospitals. However, the clinical spectrum and antimicrobial resistance differ globally. Identifying the pathogenic spectrum and variations in antibiotic resistance is crucial for controlling BSI and preventing inappropriate antibiotic use. Material and methods This retrospective observational study was conducted at the Dr. Ram Manohar Lohia Institute of Medical Sciences, Lucknow, UP, India, for one year between June 2022 and June 2023. A total of 669 adult patients' blood cultures were obtained from ICUs. Blood culture was done using a BacT/Alert 3D (BioMérieux SA, Marcy-l'Étoile, France) automated system. Identification of the bacterial as well as fungal isolates was done using matrix-assisted laser desorption ionization-time-of-flight mass spectrometry (MALDI-TOF MS), and the antimicrobial susceptibility profile was analyzed using the VITEK 2 Compact system (BioMérieux SA). Results Of the 669 blood culture samples, 213 (31.8%) showed bacterial or fungal growth. Of these 213 isolates, the most common isolate was coagulase-negative Staphylococci (21.6%), followed by Klebsiella pneumoniae (19.3%) and Acinetobacter spp. (17.8%). The majority of gram-negative bacteria were resistant to most drugs, and vancomycin and linezolid were both effective against the majority of gram-positive bacteria. Conclusion The current study found that septicemia was more frequently caused by gram-negative bacteria than by gram-positive bacteria. Blood cultures are always necessary in cases of suspected septicemia, and once the antimicrobial susceptibility profile of the pathogen causing septicemia has been determined, suitable antimicrobials should be prescribed and used to lower the antimicrobial resistance burden.

Behavioral and neuronal extracellular vesicle biomarkers associated with nicotine's enhancement of the reinforcing strength of cocaine in female and male monkeys.

While the majority of people with cocaine use disorders (CUD) also co-use tobacco/nicotine, most preclinical cocaine research does not include nicotine. The present study examined nicotine and cocaine co-use under several conditions of intravenous drug self-administration in monkeys, as well as potential peripheral biomarkers associated with co-use. In Experiment 1, male rhesus monkeys (N = 3) self-administered cocaine (0.001-0.1 mg/kg/injection) alone and with nicotine (0.01-0.03 mg/kg/injection) under a progressive-ratio schedule of reinforcement. When nicotine was added to cocaine, there was a significant leftward/upward shift in the number of injections received. In Experiment 2, socially housed female and male cynomolgus monkeys (N = 14) self-administered cocaine under a concurrent drug-vs-food choice schedule of reinforcement. Adding nicotine to the cocaine solution shifted the cocaine dose-response curves to the left, with more robust shifts noted in the female animals. There was no evidence of social rank differences. To assess reinforcing strength, delays were added to the presentation of drug; the co-use of nicotine and cocaine required significantly longer delays to decrease drug choice, compared with cocaine alone. Blood samples obtained post-session were used to analyze concentrations of neuronally derived small extracellular vesicles (NDE); significant differences in NDE profile were observed for kappa-opioid receptors when nicotine and cocaine were co-used compared with each drug alone and controls. These results suggest that drug interactions involving the co-use of nicotine and cocaine are not simply changing potency, but rather resulting in changes in reinforcing strength that should be utilized to better understand the neuropharmacology of CUD and the evaluation of potential treatments.

Characterisation of LPS+ bacterial extracellular vesicles along the gut-hepatic portal vein-liver axis.

Gut microbiome dysbiosis is a major contributing factor to several pathological conditions. However, the mechanistic understanding of the communication between gut microbiota and extra-intestinal organs remains largely elusive. Extracellular vesicles (EVs), secreted by almost every form of life, including bacteria, could play a critical role in this inter-kingdom crosstalk and are the focus of present study. Here, we present a novel approach for isolating lipopolysaccharide (LPS)+ bacterial extracellular vesicles (bEVLPS) from complex biological samples, including faeces, plasma and the liver from lean and diet-induced obese (DIO) mice. bEVLPS were extensively characterised using nanoparticle tracking analyses, immunogold labelling coupled with transmission electron microscopy, flow cytometry, super-resolution microscopy and 16S sequencing. In liver tissues, the protein expressions of TLR4 and a few macrophage-specific biomarkers were assessed by immunohistochemistry, and the gene expressions of inflammation-related cytokines and their receptors (n = 89 genes) were measured using a PCR array. Faecal samples from DIO mice revealed a remarkably lower concentration of total EVs but a significantly higher percentage of LPS+ EVs. Interestingly, DIO faecal bEVLPS showed a higher abundance of Proteobacteria by 16S sequencing. Importantly, in DIO mice, a higher number of total EVs and bEVLPS consistently entered the hepatic portal vein and subsequently reached the liver, associated with increased expression of TLR4, macrophage markers (F4/80, CD86 and CD206), cytokines and receptors (Il1rn, Ccr1, Cxcl10, Il2rg and Ccr2). Furthermore, a portion of bEVLPS escaped liver and entered the peripheral circulation. In conclusion, bEV could be the key mediator orchestrating various well-established biological effects induced by gut bacteria on distant organs.