Immune-mediated Bowel Disease: Role of Intestinal Parasites and Gut Microbiome.

Immune-mediated bowel diseases (IMBD), notably ulcerative colitis and Crohn's disease, impose a substantial global health burden due to their intricate etiology and escalating prevalence. The nexus between intestinal parasites and the gut microbiome in IMBD is a dynamic and complex field of study. Several studies have evidenced the capacity of intestinal parasites to modulate the gut microbiome, inducing alterations in microbial diversity, abundance, and metabolic activity. These changes are crucial in influencing the immune response and contributing to the development of IMBDs. Simultaneously, the gut microbiome functions as a linchpin in sustaining intestinal homeostasis and immune regulation. Dysbiosis, marked by shifts in gut microbial composition, is intricately linked to IMBD pathogenesis. Imbalances in the gut microbiota contribute to hallmark features of IMBDs, such as heightened gut permeability, chronic inflammation, and aberrant immune responses. The bidirectional interaction between intestinal parasites and the gut microbiome adds a layer of complexity to understanding IMBDs. Specific parasites, including hookworms and Necator americanus, exhibit immune downregulation and potential therapeutic applications in celiac disease. Conversely, infections with Strongyloides stercoralis and Blastocystis mirror IBD symptoms, underscoring the intricate relationship between parasites and disease pathogenesis. Further investigation is imperative to comprehensively unravel the mechanisms linking intestinal parasites and the gut microbiome in IMBD. This understanding holds the potential to pave the way for targeted therapeutic strategies aiming to restore gut microbiota homeostasis and alleviate the debilitating symptoms of these conditions. Harnessing the intricate interplay among parasites, the gut microbiome, and the host immune system may unveil novel approaches for managing and treating IMBDs.

Combinative workflow for mRNA vaccine development.

Recently, mRNA has gained a lot of attention in the field of vaccines, gene therapy, and protein replacement therapies. Herein, we are demonstrating a comprehensive approach to designing, cloning, and characterizing an antigenic cassette for the development of mRNA vaccine for COVID-19. The gene encoding the antigenic spike protein of the SARS-CoV-2 Omicron variant (B.1.1.529) was designed using the databases, characterized by in-silico tools, and assembled using overlapping oligonucleotide-based assembly by PCR. Next, the gene was cloned, mRNA was synthesized, and characterized using orthogonal approaches (Capillary electrophoresis, Sanger DNA sequencing, Next-generation sequencing, HPLC, qPCR, etc.). Furthermore, the antigen expression was monitored in-vitro using an animal cell model by western blot, flow cytometer, and surface plasmon resonance. The demonstrated approach has also been followed for developing the mRNA vaccines for various other indications such as Malaria, Herpes, Dengue, HPV, etc.

Ambient particulate matter (PM2.5) exposure contributes to neurodegeneration through the microbiome-gut-brain axis: Therapeutic role of melatonin.

Exposure to ambient particulate matter (PM2.5) has been shown to disturb the gut microbiome homeostasis and cause initiation of neuroinflammation and neurodegeneration via gut-brain bi-directional axis. Polyaromatic hydrocarbons (PAHs), which are carcinogenic and mutagenic, are important organic constituents of PM2.5 that could be involved in the microbiome-gut-brain axis-mediated neurodegeneration. Melatonin (ML) has been shown to modulate the microbiome and curb inflammation in the gut and brain. However, no studies have been reported for its effect on PM2.5-induced neuroinflammation. In the current study, it was observed that treatment with ML at 100 µM significantly inhibits microglial activation (HMC-3 cells) and colonic inflammation (CCD-841 cells) by the conditioned media from PM2.5 exposed BEAS2B cells. Further, melatonin treatment at a dose of 50 mg/kg to C57BL/6 mice exposed to PM2.5 (at a dose of 60 µg/animal) for 90 days significantly alleviated the neuroinflammation and neurodegeneration caused by PAHs in PM2.5 by modulating olfactory-brain and microbiome-gut-brain axis.

Impact of Shodhana an Ayurvedic purification process on cytotoxicity and mutagenicity of Croton tiglium Linn.

BACKGROUND: Croton tiglium Linn. (CT) which is commonly called Jaypal is used in Ayurvedic preparations like Ichhabhedi Ras, Asvakancuki Rasa. Due to its toxic contents, seeds of Croton tiglium are purified before use, by the process mentioned in classical Ayurvedic texts called Shodhana meaning purification. OBJECTIVES: The objective of the present study is to study the impact of Ayurvedic Purification process on cytotoxicity and genotoxicity of Croton tiglium Linn. MATERIALS AND METHODS: Croton tiglium Linn. Seeds were processed for Shodhana by soaking in water, heating with milk (Snehan) and later grinding in Lemon Juice (Bhavana). Aqueous and Hydroalcoholic extracts were prepared before and after purification i.e. Shodhana. Cytotoxicity of the Croton tiglium was studied against Chinese Hamster Ovary cell line by MTT assay. Ames test was performed to study the mutagenicity of the extracts in Salmonella typhi TA 98, 100 and 102 strains. Phytoconstituents were studied by using LCMS analysis. RESULTS: The results indicated decrease in cytotoxic concentration (IC50) of Croton tiglium seeds after purificationa from 3.03 mg/mL to 0.99 mg/mL in aqueous extract and 18.56 mg/mL to 5.45 mg/mL. Genotoxicity study by Ames test indicated Croton tiglium Linn. Croton tiglium Linn. Seeds are non-genotoxic in strains like S. typhi, TA 98, 100 and 102. There was change in Phytochemical profile before and after shodhana. CONCLUSION: Although both the concentrations are practically non-toxic, the decrease in cytotoxic concentration indicates Purification process as described in classical ayurvedic texts i.e. Shodhana has definitely increased the potency of the seeds of Croton tiglium Linn.

In silico evaluation of the compounds of the ayurvedic drug, AYUSH-64, for the action against the SARS-CoV-2 main protease.

BACKGROUND: Outbreak of Corona Virus Disease in late 2019 (COVID-19) has become a pandemic global Public health emergency. Since there is no approved anti-viral drug or vaccine declared for the disease and investigating existing drugs against the COVID-19. OBJECTIVE: AYUSH-64 is an Ayurvedic formulation, developed and patented by Central Council of Research in Ayurvedic Sciences, India, has been in clinical use as anti-malarial, anti-inflammatory, anti-pyretic drug for few decades. Thus, the present study was undertaken to evaluate AYUSH-64 compounds available in this drug against Severe Acute Respiratory Syndrome-Corona Virus (SARS-CoV-2) Main Protease (Mpro; PDB ID: 6LU7) via in silico techniques. MATERIALS AND METHODS: Different molecular docking software's of Discovery studio and Auto Dock Vina were used for drugs from selected AYUSH-64 compounds against SARS-CoV-2. We also conducted 100 ns period of molecular dynamics simulations with Desmond and further MM/GBSA for the best complex of AYUSH-64 with Mpro of SARS-CoV-2. RESULTS: Among 36 compounds of four ingredients of AYUSH-64 screened, 35 observed to exhibits good binding energies than the published positive co-crystal compound of N3 pepetide. The best affinity and interactions of Akuammicine N-Oxide (from Alstonia scholaris) towards the Mpro with binding energy (AutoDock Vina) of -8.4 kcal/mol and Discovery studio of Libdock score of 147.92 kcal/mol. Further, molecular dynamics simulations with MM-GBSA were also performed for Mpro- Akuammicine N-Oxide docked complex to identify the stability, specific interaction between the enzyme and the ligand. Akuammicine N-Oxide is strongly formed h-bonds with crucial Mpro residues, Cys145, and His164. CONCLUSION: The results provide lead that, the presence of Mpro- Akuammicine N-Oxide with highest Mpro binding energy along with other 34 chemical compounds having similar activity as part of AYUSH-64 make it a suitable candidate for repurposing to management of COVID-19 by further validating through experimental, clinical studies.

Oral toxicity evaluation of gokshuradi guggulu, an ayurvedic formulation.

Gokshuradi guggulu is an important classical polyherbal formulation used in Ayurvedic system of medicine for the treatment of various chronic diseases like kidney stones and diabetes. However, no scientific attempts were made to evaluate its oral toxicity. Hence, the present study evaluated the acute and 28 days repeated dose sub-acute oral toxicities of gokshuradi guggulu in rats. Gokshuradi guggulu was tested for its compliance using physicochemical and analytical parameters as per standards prescribed in Ayurvedic Pharmacopeia of India. In acute oral toxicity study, Wistar rats were orally administered a single dose of gokshuradi guggulu (2700 mg/kg) and clinical signs and mortality or moribund stage were observed for 14 days along with weekly body weight. On day 15, the rats were euthanized and the gross morphology was carried out during necropsy. In sub-acute (repeated dose) oral toxicity study, the rats were orally administered gokshuradi guggulu (270, 1350 and 2700 mg/kg) once daily up to 28 days. Clinical signs and mortality or moribund stage, weekly body weight, weekly feed and water consumptions, biochemical and hematological investigations, urine analysis, and major organ weights and histopathology were carried out. In acute and sub-acute toxicity studies, gokshuradi guggulu administration did not show any alteration in parameters or any adverse effect as compared to vehicle treated group. There was no mortality or moribund state observed in any group in both studies. Administration of gokshuradi guggulu in acute and 28 days repeated doses did not exhibit any toxicity or adverse effect at the doses used and NOAEL was found to be 2700 mg/kg.

Nephroprotective potential of Anogeissus latifolia Roxb. (Dhava) against gentamicin-induced nephrotoxicity in rats.

ETHNOPHARMACOLOGICAL RELEVANCE: Stem bark of Anogeissus latifolia Roxb. (Family: Combretaceae) is used traditionally and ethnomedicinally for correction of kidney disorders. AIM OF THE STUDY: The present study demonstrates the nephroprotective potential of stem bark of A. latifolia Roxb. MATERIALS AND METHODS: The HPTLC fingerprint and HPLC analysis were carried out to standardize the ethanolic extract of stem bark of A. latifolia (ALEE) using ellagic acid as a marker. Nephrotoxicity was induced in adult Wistar albino rats by gentamicin (100 mg/kg, intraperitoneally for 8 days) and they were treated with ALEE (100, 200 and 400 mg/kg, orally for 8 days), ellagic acid (10 mg/kg, orally for 8 days) and cystone syrup (5 ml/kg, orally), a standard reference a polyherbal formulation. Urine volume, serum and urine levels of creatinine, urea and uric acid, oxidative stress parameters (lipid peroxidation, catalase, superoxide dismutase and reduced glutathione), inflammatory markers (TNF-α and IL-6) and kidney weight along with its histological changes were studied in experimental animals. RESULTS: HPTLC, HPLC and LC-MS analysis of ALEE revealed the presence of ellagic acid and other various phytoconstituents. Administration of gentamicin caused significant increase in urine output and kidney weight, elevated biochemical, inflammatory and oxidative stress parameters as well as caused histological damage in the kidney tissue. These parameters were attenuated by the concurrent treatment with ALEE and ellagic acid. The effects were comparable to cystone. CONCLUSION: Present investigations concluded that ALEE exhibited nephroprotective potential and validated the traditional use of stem bark of A. latifolia in kidney disorders. The nephroprotective effect may be attributed to the antioxidant and anti-inflammatory phytoconstituents in ALEE.

Aldehyde-mediated bioconjugation via in situ generated ylides.

A technically simple approach for rapid, high-yielding and site-selective bioconjugation has been developed for both in vitro and cellular applications. This method involves the generation of maleimido-phosphonium ylides via 4-nitrophenol catalysis under physiological conditions followed by their Wittig reactions with aldehyde-appended biomolecules.

Gut microbiota: One of the new frontiers for elucidating fundamentals of Vipaka in Ayurveda.

With the increasing resurgence of Ayurvedic medicine in recent years, a lot of focus is laid on pharmacokinetics of herbs in arresting disease pathology. Ayurveda has enlisted some fundamentals in relation to drug pharmacokinetics, namely Rasa (perception), Virya (potency), Vipaka (postdigestive effect), Guna (properties), and Prabhava (special effect). In recent years, research has emphasized the role of gut microbiota in human health and metabolic processes. A thorough review was done to understand the role of microbiota in drug metabolism if any. The holistic mechanism of gut microbiota coincides to some extent, with the doctrines of Ayurveda in the context of pharmacodynamics and pharmacokinetics. This discussion is a thought put forth with an aim to elucidate the concept of Vipaka vis-a-vis gut microbiota functions.

Hepatitis B virus DNA polymerase gene polymorphism based prediction of genotypes in chronic HBV patients from Western India.

BACKGROUND: Hepatitis B Virus (HBV) infection is one of the major causes of liver cirrhosis, hepatocellular carcinoma and deaths due to the acute or chronic consequences worldwide. HBV is distributed into various genotypes based on nucleic acid sequence variation. OBJECTIVES: To develop a method of HBV genotyping and drug resistance interpretation using partial sequencing of polymerase gene. METHODS: This study was performed on 98 HBV infected patients' serum samples from Western India. A nested PCR protocol was designed for amplification of pol gene from HBV genome and Sanger's sequencing of the gene fragment. Sequences were aligned with HBV reference sequences for phylogenetic analysis and for characterization of genetic diversity. Drug resistance mutations were screened using HBVSeq program from Stanford University. RESULTS: Distribution of HBV genotypes showed predominance of genotype D, circulating in 76 (77.55%) patients (p < 0.05). Genotypes A and C were less prevalent and were identified in 4 (4.08%) and 18 (18.37%) patients, respectively. Anti-retroviral drug resistance mutations were not detected in any patient. CONCLUSION: A method for determination of HBV genotypes using pol gene sequencing which simultaneously detects major drug resistance mutations has been established. HBV genetic diversity may play an important role in treatment decision.

Review of Holarrhena antidysenterica (L.) Wall. ex A. DC.: Pharmacognostic, Pharmacological, and Toxicological Perspective.

Holarrhena antidysenterica (L.) Wall. ex A. DC. is a medicinal plant abundantly found in India. Its uses are mentioned in the classical Ayurvedic literature and by many folklore claims. The plant is also of extreme economic importance. Its seeds are mainly used as an antidiabetic remedy. All pharmacological and toxicological aspects of this plant are discussed in this review.

Stable and Rapid Thiol Bioconjugation by Light-Triggered Thiomaleimide Ring Hydrolysis.

Maleimide-mediated thiol-specific derivatization of biomolecules is one of the most efficacious bioconjugation approaches currently available. Alarmingly, however, recent work demonstrates that the resulting thiomaleimide conjugates are susceptible to breakdown via thiol exchange reactions. Herein, we report a new class of maleimides, namely o-CH2 NHi Pr phenyl maleimides, that undergo unprecedentedly rapid ring hydrolysis after thiol conjugation to form stable thiol exchange-resistant conjugates. Furthermore, we overcome the problem of low shelf lives of maleimide reagents owing to their propensity to undergo ring hydrolysis prior to bioconjugation by developing a photocaged version of this scaffold that resists ring hydrolysis. UV irradiation of thiol bioconjugates formed with this photocaged maleimide unleashes rapid thiomaleimide ring hydrolysis to yield the desired stable conjugates within 1 h under gentle, ice-cold conditions.

In vivo efficacy of a synthetic coumarin derivative in a murine model of aspergillosis.

Despite advances in therapeutic modalities, aspergillosis remains a leading cause of mortality. This has necessitated the identification of effective and safe antifungal molecules. In the present study, in vivo safety and antifungal efficacy of a coumarin derivative, N, N, N-Triethyl-11-(4-methyl-2-oxo-2H-benzopyran-7-yloxy)-11-oxoundecan-1-aminium bromide (SCD-1), was investigated. The maximum tolerable dose of compound was determined according to OECD 423 guidelines. The compound could be assigned to category IV of the Globally Harmonized System and its LD50 cut-off was found to be 2000 mg/kg body weight. The survival increased in Aspergillus fumigatus-infected mice treated with a dose of 200 mg/kg, orally or 100 mg/kg body weight, intraperitoneally, of SCD-1 in comparison to infected-untreated animals. The SCD-1 treatment resulted in significant reduction in colony counts in vital organs of the animals. Its protective effect was also observed on day 14 as there was marked reduction in fungal colonies. The treatment with SCD-1 also reduced the levels of serum biochemical parameters with respect to infected-untreated animals. It could be concluded that SCD-1 is a quite safe antifungal compound, which conferred dose dependent protection against experimental aspergillosis. Therefore, SCD-1 holds potential for developing new formulations for aspergillosis.