Search

Background: Increased epithelial cell death leading to compromised intestinal barrier function is a key contributor to the pathogenesis of inflammatory bowel disease. Previously published studies suggest that the nuclear bile acid receptor, farnesoid X receptor (FXR), promotes intestinal barrier function and is protective against colonic inflammation. Here, we investigated potential mechanisms involved. Methods: Mucosal inflammation was induced in mice by adding 2.5% dextran sulfate sodium (DSS) to their drinking water, either with or without daily oral gavage with the FXR agonist, obeticholic acid (OCA; 10 mg/kg). After 6 days, mice were administered FITC‐dextran (6 mg/kg) and then sacrificed 24hrs later. The severity of colonic mucosal inflammation was assessed by disease activity index (DAI) and mucosal permeability to FITC. Epithelial apoptosis was assessed by immunohistochemical imaging of cleaved caspase 3. To model the effects of cytokine-induced barrier dysfunction in vitro, we employed polarized monolayers of T84 colonic epithelial cells. Results were expressed as mean ± SEM and data were analyzed by one-way ANOVA, two-way ANOVA, and the Tukey’s post hoc test. Results: In mice, OCA treatment decreased the DSS-induced DAI score from 11.8 to 9.0 ± 0.7 (*p ≤ 0.05, n=6), increases in mucosal FITC flux by 62.3 ± 0.4% (*p ≤ 0.05, n=6), and epithelial caspase 3 cleavage by 68.1 ± 8.5% (n=6). In vitro studies revealed that treatment of T84 cells with the pro-inflammatory cytokines, IFNγ (10ng/ml) and TNFα (10ng/ml), induced apoptosis, as evidenced by increased levels of the apoptotic markers, cleaved PARP and cleaved caspase 3. However, pre-treatment of the cells with the FXR agonist, GW4064 (5μM), did not prevent cytokine-induced apoptosis. Treatment of T84 cells with IFNγ, TNFα and the apoptosis inhibitor, Q-VD-OPh, induced a necroptotic response, as evidenced by increased levels of phosphorylated RIP3 (pRIP3). Co-treatment with Q-VD-OPh also enhanced cytokine-induced transepithelial FITC flux to 3.0 ± 0.2 fold of that in control cells, whereas pre-treatment with the necroptosis inhibitor, necrostatin (200μM), reduced pRIP3 expression by 53.2 ± 3.4% (n=3) and FITC flux by 30.9 ± 0.2% (**p ≤ 0.01, n=10) of that in cells treated with Q-VD-OPh/IFNγ/TNFα alone. Similar to necrostatin, FXR activation with GW4064 inhibited both pRIP3 expression and FITC flux by 47.5 ± 9.1% (n=4) and 46.7 ± 0.6% (**p≤0.01, n=6), respectively, in this in vitro model of cytokine-induced necroptosis. Conclusion: Our studies show that FXR activation protects against intestinal inflammation, an effect that is likely due to preservation of epithelial barrier function. The protective effects of FXR activation on epithelial barrier function may be due to inhibition of necroptosis rather than apoptosis. Our data suggest that FXR represents a promising target for the development of new approaches to prevent epithelial barrier dysfunction in conditions of intestinal inflammation. Science Foundation Ireland (SFI) This is the full abstract presented at the American Physiology Summit 2023 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.

To investigate the potential anti-inflammatory and biochemical effects of Moringa peregrina leaf extracts on testosterone-induced benign prostatic hyperplasia (BPH) in rats.Six groups of rats (each group included 5 rats) were included in this study. The groups included: 1) the control group, 2) the testosterone-induced BPH group, 3) with 50 mg/kg bwt (bodyweight) oil-treated BPH, 4) with 100 mg/kg bwt. oil-treated BPH, 5) with 500mg/kg bwt. ethanol treated BPH and 6) with 1,000 mg/kg bwt. aqueous treated BPH group. Biochemical markers were measured to evaluate the effect of M. peregrina leaf extracts.Our results showed a significant improvement in the thickness of epithelial cells of the BPH glandular tissues when treated with different M. peregrina extracts (p0.05). In addition, M. peregrina extracts showed anti-inflammatory, anti-proliferative and anti-angiogenesis effects on the BPH tissues by reduction of IL-6, PCNA and VEGF-A, respectively.Our preclinical study concluded that M. peregrina leaf extracts showed a significant effect on BPH by reducing inflammation, proliferation, and angiogenic processes with no signs of toxicity.

When DNA repair is inadequate it increases the chances of the genome becoming unstable and it undergoes a malignant mutation. The deficiency of DNA repair PARP proteins may be leveraged for cancer therapy by increasing genomic instability and causing massive DNA damage in cancer cells. DNA repair components are under increased demand in cancer cells because of the continuous replication of DNA. The oncogenic loss of BRCA and an inefficient DNA repair led to cancer cells being dependent on particular DNA repair pathways, like the Poly (ADP-ribose) polymerase pathway. Breast cancer gene 1 and 2 plays a crucial role in DNA repair and genome integrity explaining how BRCA1 and BRCA2 mutations raise the menace of cancer. PARP inhibitors inhibit the base exclusion repair pathway, resulting in the buildup of unrepaired single strand breaks, which cause inflated replication forks in the S phase and subsequently the development of damaging double stranded breaks. Cells having BRCA mutations are unable to repair DNA breaks, leading to apoptosis and eventually death of cancer cells. Numerous indicators, such as a lack of homologous recombination and a high degree of replication pressure, indicate that this therapy will be very effective. Combining PARP inhibitors with chemotherapy, an immune checkpoint inhibitor, and a targeted drug is an effective strategy for combating PARP inhibitors resistance. Several PARP-based combination approaches are in preclinical and clinical development. Various clinical trials are successfully completed and some are undergoing to evaluate the efficacy of these molecules. This review will describe the current views and clinical updates on PARP inhibitors.

Ligand-based targeting of the receptors that are overexpressed explicitly on cancer cells represents an effective drug delivery approach to enhance the chemotherapeutic efficacy. Proprotein convertase subtilisin/kexin type 9 (PCSK9) which is a serine protease enzyme primarily produced by the liver cells, can potentially be used as a targeting ligand. PCSK9 binds to the LDL-r on hepatocytes' surface, leading to endocytosis and endosomal degradation. High LDL-r expression, which is believed to meet the higher demand of the cholesterol and phospholipids to build proliferating cancer cell membrane, ensures selective uptake of the PCSK9 conjugated liposomes. In the present work, the PCSK9 conjugated liposomal system was developed to deliver paclitaxel (PTX) to cancer cells. The protein was conjugated by EDC and NHS in a two-step coupling reaction to the liposomes containing COOH-PEG-COOH lipid. Conjugation was confirmed by NMR, and liposomes were further characterized by SEM and zeta sizer. PCSK9-conjugated liposomes showed high encapsulation efficiency of 69.1% with a diameter of 90.0 ± 4.9 nm. Long-term stability (30 days) study (Zeta potential: -9.88) confirmed excellent constancy and significant drug retention (58.2%). Invitro cytotoxicity and targeting efficiency was explored using MTS assay in human embryonic kidney cells (HEK293), liver hepatocellular cells (HEPG2), and a human colon cancer cell line (HCT116) for 24 h. PCSK9 conjugated liposomes exhibited significantly higher growth inhibition than the unconjugated (control) liposomes in HCT116 cell line (p

The aim of this Special Edition is to highlight the exponential work performed in the field of antimicrobial material research from the beginning of the current COVID-19 pandemic [...]

Hyper-transmissibility with decreased disease severity are typical characteristics of Omicron variant. To understand this phenomenon, we used various bioinformatics approaches to analyze randomly selected genome sequences (one each) of the Gamma, Delta, and Omicron variants submitted to NCBI from 15 to 31 December 2021. We show that: (i) Pathogenicity of SARS-CoV-2 variants decreases in the order: Wuhan > Gamma > Delta > Omicron; however, the antigenic property follows the order: Omicron > Gamma > Wuhan > Delta. (ii) Omicron Spike RBD has lower pathogenicity but higher antigenicity than other variants. (iii) Decreased disease severity by Omicron variant may be due to its decreased pro-inflammatory and IL-6 stimulation and increased IFN-γ and IL-4 induction efficacy. (iv) Mutations in N protein are associated with decreased IL-6 induction and human DDX21-mediated increased IL-4 production in Omicron. (v) Due to mutations, the stability of S, M, N, and E proteins decreases in the order: Omicron > Gamma > Delta > Wuhan. (vi) Stronger Spike RBD-hACE2 binding in Omicron is associated with increased transmissibility. However, the lowest stability of the Omicron Spike protein makes Spike RBD-hACE2 interaction weak for systemic infection and for causing severe disease. Finally (vii), the highest instability of Omicron E protein may also be associated with decreased viral maturation and low viral load leading to less severe disease and faster recovery. Our method may be used for other similar viruses, and these findings will contribute to the understanding of the dynamics of SARS-CoV-2 variants and the management of emerging variants.

Several hypotheses have been presented on the origin of severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) from its identification as the agent causing the current coronavirus disease 19 (COVID-19) pandemic. So far, no solid evidence has been found to support any hypothesis on the origin of this virus, and the issue continue to resurface over and over again. Here we have unfolded a pattern of distribution of several mutations in the SARS-CoV-2 proteins in 24 geo-locations across different continents. The results showed an evenly uneven distribution of the unique protein variants, distinct mutations, unique frequency of common conserved residues, and mutational residues across these 24 geo-locations. Furthermore, ample mutations were identified in the evolutionarily conserved invariant regions in the SARS-CoV-2 proteins across almost all geo-locations studied. This pattern of mutations potentially breaches the law of evolutionary conserved functional units of the beta-coronavirus genus. These mutations may lead to several novel SARS-CoV-2 variants with a high degree of transmissibility and virulence. A thorough investigation on the origin and characteristics of SARS-CoV-2 needs to be conducted in the interest of science and for the preparation of meeting the challenges of potential future pandemics.

Several chemicals, such as dextran sulfate sodium (DSS), oxazolone, acetic acid, and trinitrobenzene sulphonic acid (TNBS), have been used for establishing animal models of ulcerative colitis. These animal models help us to study or explore several factors involved in the etiology or pathogenesis of ulcerative colitis. They are also useful tools to design and develop effective drug delivery strategies. DSS is the most widely used tool to induce colitis in animals. The model of ulcerative colitis developed by this method effectively mimics the colitis condition in humans. The amount of DSS in drinking water can be adjusted to control the severity of colitis, such as acute or chronic inflammation. However, a miscalculation in the amount of DSS produces severe inflammation, which may lead to the death of mice. DSS has been shown to rupture the epithelial lining and induce infiltration of inflammatory markers such as TNF, interferons, and interleukins. The current study aims to study the effects of different amounts of DSS on weight loss, changes in colon length, and histological scoring. Furthermore, the main objective of this study was to find an optimum concentration of DSS to establish a mouse model for ulcerative colitis. Based on the disease index, weight loss, bleeding, histological studies, and colon length, 2.5% w/v DSS for 7 days in water was found to be adequate for the DSS-induced colitis model for a moderate level of colitis, and 3.5% w/v DSS could be used to study severe experimental colitis.

Open reading frame 8 (ORF8) shows one of the highest levels of variability among accessory proteins in Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2), the causative agent of Coronavirus Disease 2019 (COVID-19). It was previously reported that the ORF8 protein inhibits the presentation of viral antigens by the major histocompatibility complex class I (MHC-I), which interacts with host factors involved in pulmonary inflammation. The ORF8 protein assists SARS-CoV-2 in evading immunity and plays a role in SARS-CoV-2 replication. Among many contributing mutations, Q27STOP, a mutation in the ORF8 protein, defines the B.1.1.7 lineage of SARS-CoV-2, engendering the second wave of COVID-19. In the present study, 47 unique truncated ORF8 proteins (T-ORF8) with the Q27STOP mutations were identified among 49,055 available B.1.1.7 SARS-CoV-2 sequences. The results show that only one of the 47 T-ORF8 variants spread to over 57 geo-locations in North America, and other continents, which include Africa, Asia, Europe and South America. Based on various quantitative features, such as amino acid homology, polar/non-polar sequence homology, Shannon entropy conservation, and other physicochemical properties of all specific 47 T-ORF8 protein variants, nine possible T-ORF8 unique variants were defined. The question as to whether T-ORF8 variants function similarly to the wild type ORF8 is yet to be investigated. A positive response to the question could exacerbate future COVID-19 waves, necessitating severe containment measures.

Phylogeny is a statistical approach displaying the evolutionary history of a genetically related group of organisms with the fundamental prerequisite of the utilization of a significant sample size that could represent the whole population under investigation. Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) phylogeny analyses are based on a single isolate of BatCoVs, which is not a sufficient representation of genetically related CoVs. For instance, the unique Bat-CoV RaTG13 sequence that is genetically associated with SARS-CoV-2 was isolated from Yunnan, China, in 2013. To date, no other RaTG13 sequence has been obtained in a different time (temporal), place (spatial), or other host condition. Data scarcity of Bat-CoVs sequences raises concern on the several fundamental experimental and biostatistical aspects, e.g. repeatability of the sequences and intraspecies variation of critical genes, such as the receptor-binding domain of Spike protein. Sunda pangolin has been proposed as the intermediate host and source of SARS-CoV-2, but no Pangolin-CoV isolates have been isolated in Southeast Asia, where Sunda pangolins inhabit. Most Pangolin-CoVs were isolated from deceased pangolins, that were captured during illegal animal trafficking into China, hence raising questions about the reliability and quality of such isolates. Pangolin-CoV sampling problems are also evident in the deposited sequences that are of sub-standard quality. Therefore, there is urgent need for survey the Bat-CoVs and possible intermediate hosts, such as pangolins and civets in Southeast Asia. These surveys are required to investigate the genomic source of SARS-CoV-2 and assess possible future risks for new outbreaks. The current SARS-CoV-2 phylogeny with unacceptably limited numbers of Bat-CoVs and Pangolin-CoVs sequences not sufficient and technically not appropriate for reliable phylogenic analysis.

Asthma is a common, heterogeneous and serious disease, its prevalence has steadily risen in most parts of the world, and the condition is often inadequately controlled in many patients. Hence, there is a major need for new therapeutic approaches. Mild-to-moderate asthma is considered a T-helper cell type-2-mediated inflammatory disorder that develops due to abnormal immune responses to otherwise innocuous allergens. Prolonged exposure to allergens and persistent inflammation results in myofibroblast infiltration and airway remodelling with mucus hypersecretion, airway smooth muscle hypertrophy, and excess collagen deposition. The airways become hyper-responsive to provocation resulting in the characteristic wheezing and obstructed airflow experienced by patients. Extensive research has progressed the understanding of the underlying mechanisms and the development of new treatments for the management of asthma. Here, we review the basis of the disease, covering new areas such as the role of vascularisation and microRNAs, as well as associated potential therapeutic interventions utilising reports from animal and human studies. We also cover novel drug delivery strategies that are being developed to enhance therapeutic efficacy and patient compliance. Potential avenues to explore to improve the future of asthma management are highlighted.

The coronavirus disease 2019 (COVID-19) is caused by the Severe Acute Respiratory Syndrome Coronavirus-2 (SARS- CoV-2) with an estimated fatality rate of less than 1%. The SARS-CoV-2 accessory proteins ORF3a, ORF6, ORF7a, ORF7b, ORF8, and ORF10 possess putative functions to manipulate host immune mechanisms. These involve interferons, which appear as a consensus function, immune signaling receptor NLRP3 (NLR family pyrin domain-containing 3) inflammasome, and inflammatory cytokines such as interleukin 1β (IL-1β) and are critical in COVID-19 pathology. Outspread variations of each of the six accessory proteins were observed across six continents of all complete SARS-CoV-2 proteomes based on the data reported before November 2020. A decreasing order of percentage of unique variations in the accessory proteins was determined as ORF3a > ORF8 > ORF7a > ORF6 > ORF10 > ORF7b across all continents. The highest and lowest unique variations of ORF3a were observed in South America and Oceania, respectively. These findings suggest that the wide variations in accessory proteins seem to affect the pathogenicity of SARS-CoV-2.

Lung cancer, a group of destructive malignant tumors, is a leading cause of new cancer cases and cancer-associated deaths. The application of nanoemulsions is an attractive platform for cancer therapy, particularly due to their favorable properties to solubilize hydrophobic chemotherapeutics, thermodynamic stability, biocompatibility, and targetability. A multifunctional role for nanoemulsions could become possible for their submicron dispersed colloids in the continuous phase, which further aids in overcoming biological barriers associated with conventional chemotherapeutic drug delivery to reach the target site of lung cancer. Surface-engineered nanoemulsion with incorporated chemotherapies has the potential to revolutionize lung cancer therapy by tailoring it according to the requirement to target the microenvironment of cancer cells for improved therapy. Several nanoemulsion formulations are in different stages of clinical phases, however, to date, there is no approved nanoemulsion marketed for lung cancer. In this chapter, the authors focus on different approaches to nanoemulsion use for selective and improved lung cancer targeting and emphasize the underlying challenges in lung cancer therapy.