Plasma and urine metabolite profiling reveals the protective effect of Clinacanthus nutans in an ovalbumin-induced anaphylaxis model: ¹H-NMR metabolomics approach

The present study sought to identify the key biomarkers and pathways involved in the induction of allergic sensitization to ovalbumin and to elucidate the potential anti-anaphylaxis property of Clinacanthus nutans (Burm. f.) Lindau water leaf extract, a Southeast Asia herb in an in vivo ovalbumin-induced active systemic anaphylaxis model evaluated by 1H-NMR metabolomics. The results revealed that carbohydrate metabolism (glucose, myo-inositol, galactarate) and lipid metabolism (glycerol, choline, sn-glycero-3-phosphocholine) are the key requisites for the induction of anaphylaxis reaction. Sensitized rats treated with 2000 mg/kg bw C. nutans extract before ovalbumin challenge showed a positive correlation with the normal group and was negatively related to the induced group. Further 1H-NMR analysis in complement with Kyoto Encyclopedia of Genes and Genomes (KEGG) reveals the protective effect of C. nutans extract against ovalbumin-induced anaphylaxis through the down-regulation of lipid metabolism (choline, sn-glycero-3-phosphocholine), carbohydrate and signal transduction system (glucose, myo-inositol, galactarate) and up-regulation of citrate cycle intermediates (citrate, 2-oxoglutarate, succinate), propanoate metabolism (1,2-propanediol), amino acid metabolism (betaine, N,N-dimethylglycine, methylguanidine, valine) and nucleotide metabolism (malonate, allantoin). In summary, this study reports for the first time, C. nutans water extract is a potential anti-anaphylactic agent and 1H-NMR metabolomics is a great alternative analytical tool to explicate the mechanism of action of anaphylaxis

Anti-allergic effect of Clinacanthus nutans aqueous extract: protection against IgE-mediated passive systemic anaphylaxis

Introduction: Anaphylaxis is a serious, rapid and potentially life-threatening allergic response involving IgE or IgG. Clinacanthus nutans, a small native shrub found in tropical Asia possess analgesic, anti-inflammatory and anti-viral activities and traditionally used for skin rashes, insect and snake-bites. In Thailand, alcoholic C. nutans extracts has been used topically for skin rashes, a symptom of allergy. Aim: To justify that C. nutans can treat skin rashes; this study investigated the anti-allergenicity of C. nutans extracts. Methods: Cytotoxicity of C. nutans extracts was evaluated by MTT on RBL-2H3. The most active C. nutans extract was determined by IgE-mediated mast cell degranulation. Acute toxicity of C. nutans aqueous extract was evaluated using female Sprague Dawley rats at 5000 mg/kg. Anti-allergenicity of C. nutans aqueous extract was studied by IgE-mediated passive systemic anaphylaxis (PSA). The release of preformed mediator (β-hexosaminidase) as well as newly synthesized mediators (TNF-α, IL-4 and LTC4) was evaluated. Results: C. nutans extracts were not cytotoxic up to 1 mg/ml (ethanolic) and 6 mg/ml (aqueous). In vitro, C. nutans aqueous extract was able to inhibit the release of preformed mediators but not newly synthesized mediators at 5 mg/ml. The ethanolic extracts were not able to inhibit all mediators tested. At 5000 mg/kg, C. nutans aqueous extract was non-toxic to the rats; no significant difference observed haematologically and biochemically. In vivo, C. nutans aqueous extract did not inhibit mediators of IgE-mediated PSA at 500 mg/kg and 2000 mg/kg. Conclusion: C. nutans aqueous extract was most active but could not inhibit mediators of IgE-mediated PSA. As anaphylaxis could be mediated by IgE orIgG, we postulate that C. nutans aqueous extract may exhibit its anti-allergenicity in IgG-mediated pathway

Phytochemical diversity of Clinacanthus nutans extracts and their bioactivity correlations elucidated by NMR based metabolomics

Clinacanthus nutans is a medicinal herb and a traditional remedy for herpes viral infection, cancer, and diabetes mellitus. Despite its popular use, there is limited information on the chemical constituents and their relationship with the bioactivities of the herb. The choice of drying and extraction methods will greatly influence the metabolite profile and the bioactivities of an herbal extract. In order to maximize retention of the original chemical profile of the herb and quality assurance, optimization of processing methods is needed. Using nuclear magnetic resonance (NMR) based metabolomics approach, we have carried out a discriminative analysis of the metabolite profiles of the leaves and stems of the herb when different drying (air, oven, and freeze) and extraction (soaking and sonication) methods were used and correlated the metabolite profiles with the total phenolic content (TPC), antioxidant and α-glucosidase inhibitory activities. Identification of primary and secondary metabolites was performed using 1D- and 2D-NMR techniques as well as ultra-performance liquid chromatography–tandem mass spectrometry (UPLC–MS/MS). Results showed that the leaf extracts, which were richer in phenolic compounds and terpenoids, showed significantly higher bioactivities compared to the stem extracts. Several newly reported compounds in the herb, identified using tandem mass spectrometry, included gendarucin A, a gendarucin A isomer, 3,3-di-O-methylellagic acid, ascorbic acid, and two isomeric oxoprolinates. From the NMR metabolomics analysis, the PLS biplot model indicated that the presence of some phenolics compounds, terpenoids, and sulfur-containing glucosides in the oven and air dried leaf extracts are the main components responsible for the antioxidant and α-glucosidase inhibitory activities. This study has provided additional information on the chemistry and biology of the herb that may be useful in future development phytomedicinal preparations of C. nutans

Identification of Soluble Mediators in IgG-Mediated Anaphylaxis via Fcγ Receptor: A Meta-Analysis

Background: Anaphylaxis is an acute and life-threatening allergic response. Classically and most commonly, it can be mediated by the crosslinking of allergens to immunoglobulin E (IgE)- high affinity IgE receptor (FcεRI) complex found mostly on mast cells. However, there is another pathway of anaphylaxis that is less well-studied. This pathway known as the alternative pathway is mediated by IgG and its Fc gamma receptor (Fcγ). Though it was not documented in human anaphylaxis, a few studies have found that IgG-mediated anaphylaxis can happen as demonstrated in rodent models of anaphylaxis. In these studies, a variety of soluble mediators were being evaluated and they differ from each study which causes confusion in the suitability, and reliability of choice of soluble mediators to be analyzed for diagnosis or therapeutic purposes. Hence, the objective of this meta-analysis is to identify the potential soluble mediators that are involved in an IgG-mediated anaphylaxis reaction.Methods: Studies related to IgG-mediated anaphylaxis were sourced from five search engines namely PubMed, Scopus, Ovid, Cochrane Library, and Center for Agricultural Bioscience International (CABI) regardless of publication year. Relevant studies were then reviewed based on specific inclusion factors. The means and standard deviations of each soluble mediator studied were then extracted using ImageJ or Get Data Graph Digitiser software and the data were subjected to meta-analysis.Results: From our findings, we found that histamine, serotonin, platelet activating factor (PAF), β-hexosaminidase, leukotriene C4 (LTC4), mucosal mast cell protease-1 (MMCP-1), interleukins (IL)-4,−6, and−13; tumor necrosis factor alpha (TNF-α), and macrophage inflammatory protein-1α (MIP-1α) were often being analyzed. Out of these soluble mediators, histamine, PAF, β-hexosaminidase, IL-6, and−13, MIP-1α and TNF-α were more significant with positive effect size and p < 0.001. As study effect was relatively small, we performed publication bias and found that there was publication bias and this could be due to the small sample size studied.Conclusion: As such, we proposed that through meta-analysis, the potential soluble mediators involved in rodent IgG-mediated anaphylaxis to be histamine, PAF, β-hexosaminidase, IL-6 and−13 and MIP-1α, and TNF-α but will require further studies with larger sample size

Hematological, biochemical, histopathological and 1H-NMR metabolomics application in acute toxicity evaluation of clinacanthus nutans water leaf extract

The present study aims for the first time to provide the in vivo acute toxicological profile of the highest dose of Clinacanthus nutans (Burm. f.) Lindau water leaf extract according to the Organization for economic co-operation and development (OECD) 423 guidelines through conventional toxicity and advanced proton nuclear magnetic resonance (1H-NMR) serum and urinary metabolomics evaluation methods. A single dose of 5000 mg/kg bw of C. nutans water extract was administered to Sprague Dawley rats, and they were observed for 14 days. Conventional toxicity evaluation methods (physical observation, body and organ weight, food and water consumption, hematology, biochemical testing and histopathological analysis) suggested no abnormal toxicity signs. Serum 1H-NMR metabolome revealed no significant metabolic difference between untreated and treated groups. Urinary 1H-NMR analysis, on the other hand, revealed alteration in carbohydrate metabolism, energy metabolism and amino acid metabolism in extract-treated rats after 2 h of extract administration, but the metabolic expression collected after 24 h and at Day 5, Day 10 and Day 15 indicated that the extract-treated rats did not accumulate any toxicity biomarkers. Importantly, the outcomes further suggest that single oral administration of up to 5000 mg/kg bw of C. nutans water leaf extract is safe for consumption

Minimal information for studies of extracellular vesicles (MISEV2023): From basic to advanced approaches

Extracellular vesicles (EVs), through their complex cargo, can reflect the state of their cell of origin and change the functions and phenotypes of other cells. These features indicate strong biomarker and therapeutic potential and have generated broad interest, as evidenced by the steady year-on-year increase in the numbers of scientific publications about EVs. Important advances have been made in EV metrology and in understanding and applying EV biology. However, hurdles remain to realising the potential of EVs in domains ranging from basic biology to clinical applications due to challenges in EV nomenclature, separation from non-vesicular extracellular particles, characterisation and functional studies. To address the challenges and opportunities in this rapidly evolving field, the International Society for Extracellular Vesicles (ISEV) updates its 'Minimal Information for Studies of Extracellular Vesicles', which was first published in 2014 and then in 2018 as MISEV2014 and MISEV2018, respectively. The goal of the current document, MISEV2023, is to provide researchers with an updated snapshot of available approaches and their advantages and limitations for production, separation and characterisation of EVs from multiple sources, including cell culture, body fluids and solid tissues. In addition to presenting the latest state of the art in basic principles of EV research, this document also covers advanced techniques and approaches that are currently expanding the boundaries of the field. MISEV2023 also includes new sections on EV release and uptake and a brief discussion of in vivo approaches to study EVs. Compiling feedback from ISEV expert task forces and more than 1000 researchers, this document conveys the current state of EV research to facilitate robust scientific discoveries and move the field forward even more rapidly

Retrospective evaluation of whole exome and genome mutation calls in 746 cancer samples

Funder: NCI U24CA211006Abstract: The Cancer Genome Atlas (TCGA) and International Cancer Genome Consortium (ICGC) curated consensus somatic mutation calls using whole exome sequencing (WES) and whole genome sequencing (WGS), respectively. Here, as part of the ICGC/TCGA Pan-Cancer Analysis of Whole Genomes (PCAWG) Consortium, which aggregated whole genome sequencing data from 2,658 cancers across 38 tumour types, we compare WES and WGS side-by-side from 746 TCGA samples, finding that ~80% of mutations overlap in covered exonic regions. We estimate that low variant allele fraction (VAF < 15%) and clonal heterogeneity contribute up to 68% of private WGS mutations and 71% of private WES mutations. We observe that ~30% of private WGS mutations trace to mutations identified by a single variant caller in WES consensus efforts. WGS captures both ~50% more variation in exonic regions and un-observed mutations in loci with variable GC-content. Together, our analysis highlights technological divergences between two reproducible somatic variant detection efforts

Phytochemical profile and anti-allergy properties of Clinacanthus nutans (Burm.F) Lindau extract revealed by ¹1H-NMR-based metabolomics

Food allergy is an abnormal immunological response arises when a person's immune system wrongly interprets a food as a harmful foreign invader. Although the existing synthetic derived anti-allergy drugs provide effective treatment, the possible side effects on human have drawn the researchers’ attention to seek alternatives from nature source. The present study aimed to profile the metabolites and investigate the pharmacological potency of Clinacanthus nutans, a local traditional herb and vegetable, in managing food allergy using proton nuclear magnetic resonance (1HNMR) metabolomics approach. The first and second parts of the study focused on evaluating the effect of different plant parts, dryings, binary extraction solvent system (ethanol: water ratio) and extraction methods on the phytochemical composition and bioactivity of the plant using metabolomics approach. Subsequently, the optimised plant extract was assessed for its acute toxicity effect on Sprague Dawley rats at a single oral dose of 5000 mg/kg body weight (BW) using both conventional (visual behavioural observation, hematological and biochemical tests) and advanced (serumurinary 1H-NMR metabolomics) toxicity evaluation methods. The optimal plant extract was further evaluated for its anti-allergy effect in a dose dependent manner (125, 500 and 2000 mg/kg BW rat) using in vivo ovalbumin induced active systemic anaphylaxis (OVA-ASA) rat model. From the experiment, water-sonication prepared C. nutans air dried leaf was determined as the optimal extract as it had the highest TPC (51.44 ± 0.63 mg GAE/g of extract) and exhibited the best NO inhibitory activity (IC50: 190.43 ± 12.26 μg/mL) without adverse effect on the RAW cell viability. The analysis further suggested the potential NO inhibitors in the extract were clinacoside A and B, clinamide A, B and C, phytosterols, lupeol, orientin, isoorientin, valine, alanine, acetic acid, and lactic acid. The acute toxicity test showed that the extract-treated rats behaved normal with no observed morbidity, mortality or apparent signs of toxicity. Therefore, the LD50 of C. nutans extract is likely to be above 5000 mg/kg BW. Consequently, the anti-allergy properties of the optimal C. nutans extract was evaluated. A stronger positive correlation of the high dose extract-treated group (2000 mg/kg BW) with the normal group than with the ovalbumin-induced group was observed. The metabolic pathway further suggested the anti-allergy effect of C. nutans may be ascribed to its ability in modulating the carbohydrate metabolism, energy metabolism, lipid metabolism, amino acid metabolism and nucleotide metabolism of sensitised rats. This study is the first report on the potency of C. nutans as an antiallergy agent. The finding of this study therefore, paves a new way for future functional food or drug development in mitigating allergy reaction and lays the basis for any metabolomics related study

A Comprehensive Review on Phytochemistry and Pharmacological Activities of Clinacanthus nutans (Burm.f.) Lindau

Clinacanthus nutans (Burm.f.) Lindau (Acanthaceae), commonly known as Sabah snake grass, is a vegetable and a well-known herb that is considered an alternative medicine for insect bites, skin rashes, herpes infection, inflammation, and cancer and for health benefits. Current review aims to provide a well-tabulated repository of the phytochemical screening, identification and quantification, and the pharmacological information of C. nutans according to the experimental design and the plant preparation methods which make it outstanding compared to existing reviews. This review has documented valuable data obtained from all accessible library databases and electronic searches. For the first time we analyzed the presence of flavonoids, triterpenoids, steroids, phytosterols, and glycosides in C. nutans based on the results from phytochemical screening which are then further confirmed by conventional phytochemical isolation methods and advanced spectroscopic techniques. Phytochemical quantification further illustrated that C. nutans is a good source of phenolics and flavonoids. Pharmacological studies on C. nutans revealed that its polar extract could be a promising anti-inflammation, antiviral, anticancer, immune and neuromodulating, and plasmid DNA protective agent; that its semipolar extract could be a promising antiviral, anticancer, and wound healing agent; and that its nonpolar extract could be an excellent anticancer agent