Nuclear magnetic resonance metabolomics approach in chemical and protective evaluations of Orthosiphon stamineus Benth. leaf extracts on cisplatin-induced nephrotoxicity

Orthosiphon stamineus (OS), locally known in Malaysia as ‘Misai Kucing’, is a herbaceous shrub belonging to the family Lamiaceae. Dried leaves of OS is gaining wide acceptance and marketed in the form of herbal tea, known as Java tea, owing to its traditional and scientific claims on various health benefits. OS has been a wellknown renoprotective agent primarily due to its diuretic potential. This research investigated the effects of commonly employed drying methods of OS leaves on their chemical constituent profile, and in vivo biological properties of the protective role in cisplatin induced nephrotoxicity using rats, through Nuclear Magnetic Resonance (NMR) metabolomics approach. The NMR spectra of rat urine and the OS leaf extracts were analysed and correlated using multivariate data analysis techniques employing metabolomics platform. The 1H NMR metabolite profiling of aqueous extract of OS leaves resulted in the identification of 31 metabolites. The presence of biologically active secondary metabolites including phenylpropanoids such as caffeic acid, protocatechuic acid, chlorogenic acid, flavonoids such as luteolin and apigenin, gallic acid and orthosiphol derivatives were confirmed by J resolved NMR technique. The HPLC - MS/MS analysis further confirmed the presence of these secondary metabolites. Metabolite fingerprinting in combination with multivariate analysis has successfully differentiated the three differently dried (Freeze, microwave and shade) OS leaves and established that the levels of 15 metabolites were varied significantly between the samples. The shade drying method retained maximum secondary metabolites followed by the microwave, while freeze drying retained the least. Assessment of the main beneficial properties, such as antioxidant, total phenolic and flavonoid contents of any tea preparation, confirmed that all the differently dried Java tea leaves gave good antioxidant activity, with the shade dried leaves recorded the highest level with an IC50 of 48.09 μg/mL. The chemical constituents correlated to the high antioxidant activity of the shade dried leaves were extracted from a Partial Least Square regression (PLS) model. In addition, the toxicity profile of the microwave dried OS leaves was investigated through acute oral toxicity test in Sprague Dawley (SD) rats of both sexes, whereby, the no-observed-adverse-effect level (NOAEL) of aqueous, 50% ethanolic and ethanolic extracts of the microwave dried OS was determined as 5000 mg/kg body weight/day. Thus, it is presumed that the microwave dried leaves are safe to be used as an oral health supplement. Cisplatin is an anticancer drug, which induces nephrotoxicity in a long term use. Metabolomic analysis of the rats’ urine revealed the involvement of a total of 17 biochemical markers from TCA cycle, carbohydrate, amino acid, and polyamine metabolic pathways in cisplatin nephrotoxicity. To the best of knowledge, 6 of the 17 involved metabolites are newly established in this study. In order to evaluate the protective efficacy of OS in cisplatin nephrotoxicity, shade and microwave dried OS extracts were administered at doses of 100, 200 and 400 mg/kg body weight to rats. The results suggested the dose independency of the extracts. Treatment with 50% aqueous ethanolic extract of shade dried OS leaves (OSFS) exhibited moderate ameliorative effect observed through a statistically significant reduction in the levels of 8 biomarkers. It was also revealed that the aqueous extract of the shade dried leaves (OSAS) exhibited slightly deteriorative activity via disturbance in the energy metabolism and gut microflora. The higher concentration of the secondary metabolites such as caffeic acid, chlorogenic acid, protocatechuic acid and orthosiphol in OSFS could be correlated to the ameliorative activity as revealed from a Principal Component Analysis (PCA) between OSAS and OSFS. A prediction model on nephroprotective effect of OS was constructed through PLS regression analysis. Thus, the impact of different drying techniques on chemical constituents of OS leaves was established. The metabolomics approach has proved to be successful in shedding light to the even minute variations in the biological profiles of the low intensity metabolites involved in the renal toxicity caused by cisplatin. A global comprehensive view of the OS effect in cisplatin toxicity was successfully profiled and correlated

Anti inflammatory effect of rhizome of Curcuma longa. Linn, in Albino rats by the method of Carrageenin induced paw oedema

Background: Curcuma longa or turmeric is a popular Ayurvedic herb, traditionally used for various inflammatory conditions including rheumatoid arthritis and spondylitis. Turmeric which containing phytochemical ingredient curcumin is used in India for centuries as a topical anti inflammatory agent. Many of the currently used anti inflammatory agents like NSAID’s and glucocorticoids have many undesirable adverse effects, especially when they are used for long period. In the present study, Curcuma longa, a plant belonging to the Zingiberaceae family was chosen for investigating its anti-inflammatory effects.Methods: The rhizomes of Curcuma longa were collected locally. The extract was prepared by soxlet extraction with 50% ethanol. Albino rats of Wistar strain (170-250grams) obtained from the animal house of medical college Thiruvananthapuram were used. Aspirin was purchased from sigma Labs, Mumbai. Anti-inflammatory effect of the extract was done in rats by the method of Carrageenin induced paw oedema.Results: The ethanolic extract of Curcuma longa inhibited the development of oedema at the end of 3 hours. The anti-inflammatory activity exhibited by the extract was dose dependent and statistically significant at dose levels of 1000-mg/kg and comparable to that of standard drug used Aspirin.Conclusions: The present study with extract of Curcuma longa revealed that it has significant anti-inflammatory activity

Anti-ulcer effect of rhizome of Curcuma longa Linn, by the method of pyloric ligation

Background: Substantial part of the world population has been known for a long time to suffer from peptic ulcer disease. In the present study, Curcuma longa, a plant belonging to the Zingiberaceae family was chosen for investigating its anti-ulcer properties.Methods: The rhizomes of Curcuma longa were collected locally. The extract was prepared by soxhlet extraction with 50% ethanol. Albino rats of Wistar strain (120-200 grams) obtained from the animal house of medical college Thiruvananthapuram were used. Ranitidine was collected from Kerala Sate Drugs and Pharmaceutical LTD Alapuzha. Antiulcer study in rats were done using the method of pyloric ligation.Results: Antiulcer study in rats using the method of pyloric ligation, extract of Curcuma longa in 1000mg/Kg dose levels exhibited significant protection against shay ulceration. The results were comparable to that of standard drug Ranitidine.Conclusions: The present study with extract of Curcuma longa revealed that it has significant anti-ulcer activity

GUILLAIN-BARRE SYNDROME FOLLOWING CHADOX1 NCOV-19 COVID-19 VACCINATION AT AN ADR MONITORING CENTER IN A TERTIARY CARE HOSPITAL, KOZHIKODE: A CASE SERIES.

Guillain barre syndrome (GBS) is a rare but fatal autoimmune disease affecting the nervous system. The occurrence of GBS after COVID vaccination is rare and its exact cause is still unknown. Hence, here, we are trying to describe the pattern of GBS following the first dose of COVID-19 vaccination as a case series. The retrospective case series study was carried out by analyzing the serious AEFI (Adverse Events Following Immunization) case notification form reported by health-care professionals to ADR Monitoring Center during the period of March 2021–September 2021.The purpose of these case reports is to ensure proper surveillance methods to monitor the safety of COVID vaccination and to promote further researches which is required to determine the possible link between GBS and COVID-19 vaccination

Formulation and Evaluation of Spray-Dried Esomeprazole Magnesium Microspheres

Purpose: To prepare and evaluate in vitro mucoadhesive esomeprazole magnesium microspheres for the treatment of Zollinger Ellison syndrome.Methods: The microspheres were prepared by spray drying technique using locust bean and xanthan gums as polymers. Esomeprazole magnesium was entrapped in the microspheres at various polymer/cross-linking ratios. Glutaraldehyde was used to cross-link the gums. The microspheres wereevaluated for their micromeritic properties and in vitro release. as well as by Fourier transform infrared (FTIR), differential scanning calorimetry (DSC) and scanning electron microscopy (SEM).Results: The microspheres were discrete, spherical, and showed good drug entrapment efficiency (60.5 - 92.3 %). FTIR and DSC results indicate that the drug was compatible with the polymers used. Amongst all the formulations, F6 (drug:locust bean gum:xanthan gum, 1:2:2) showed the most suitable sustained release properties with 99.8 % of drug released at the end of 12 h.Conclusion: Microspheres prepared using locust bean and xanthan gums can be used as a sustained release delivery system for esomeprazole magnesium.Keywords: Spray drying, Microspheres, Esomeprazole magnesium, Sustained releas

Untargeted Metabolic Fingerprinting Reveals Impact of Growth Stage and Location on Composition of Sea Buckthorn (Hippophaë Rhamnoides) Leaves

Sea buckthorn (Hippophaë rhamnoides) is increasingly cultivated to produce raw materials for food and nutraceuticals. There is little knowledge on composition of sea buckthorn leaves (SBLs) and the key factors influencing the composition. This research aims to unravel the metabolic profile of SBLs and the effects of cultivar, location and stage of growth, and climatic conditions on the metabolic profile of SBLs. Leaves of two sea buckthorn cultivars grown in the south and north of Finland during two consecutive growth seasons were studied using untargeted nuclear magnetic resonance (NMR) metabolomics. The highest variance in the metabolic profile was linked to the growth stage, wherein leaves from the first 7 weeks of harvest were characterized with higher abundance of polyphenols, while relatively higher abundance of carbohydrates and sugars was observed in the later weeks. The growth location attributed for the second highest variation, wherein the north–south comparison identified fatty acids and sugars as discriminatory metabolites, and the potential association of metabolome to natural abiotic stressors was revealed. An inverse correlation between carbohydrate/sugar content as well as fatty acids of higher carbon chain length with the temperature variables was evident. The supervised chemometric models with high sensitivity and specificity classified and predicted the samples based on growth stage and location, and cultivar. Nontargeted NMR-metabolomics revealed the metabolic profile of SBLs and their variation associated with various biotic and abiotic factors. Cultivar and growth stage are key factors to consider when harvesting SBLs for use in food and nutraceuticals.Practical ApplicationGlobally, sea buckthorn cultivation has been rapidly increasing due to the known health-promoting benefits of the berries and leaves of the plant. The current research obtained new comprehensive information on the compositional profile of sea buckthorn leaves as well as the impact of major contributory factors, such as cultivars, the advancement of growth stage, geographical location, and weather parameters. The findings of this research provide new knowledge and guidance for plant breeding, cultivation and commercial utilization of sea buckthorn leaves as raw materials for food, feed, and nutraceuticals.</p

Prebiotic Xylo-Oligosaccharides Ameliorate High-Fat-Diet-Induced Hepatic Steatosis in Rats

Understanding the importance of the gut microbiota (GM) in non-alcoholic fatty liver disease (NAFLD) has raised the hope for therapeutic microbes. We have shown that high hepatic fat content associated with low abundance of Faecalibacterium prausnitzii in humans and, further, the administration of F. prausnitzii prevented NAFLD in mice. Here, we aimed at targeting F. prausnitzii by prebiotic xylo-oligosaccharides (XOS) to treat NAFLD. First, the effect of XOS on F. prausnitzii growth was assessed in vitro. Then, XOS was supplemented or not with high (HFD, 60% of energy from fat) or low (LFD) fat diet for 12 weeks in Wistar rats (n = 10/group). XOS increased F. prausnitzii growth, having only a minor impact on the GM composition. When supplemented with HFD, XOS ameliorated hepatic steatosis. The underlying mechanisms involved enhanced hepatic β-oxidation and mitochondrial respiration. Nuclear magnetic resonance (1H-NMR) analysis of cecal metabolites showed that, compared to the HFD, the LFD group had a healthier cecal short-chain fatty acid profile and on the HFD, XOS reduced cecal isovalerate and tyrosine, metabolites previously linked to NAFLD. Cecal branched-chain fatty acids associated positively and butyrate negatively with hepatic triglycerides. In conclusion, XOS supplementation can ameliorate NAFLD by improving hepatic oxidative metabolism and affecting GM

NMR-based metabolomics approach on optimization of malolactic fermentation of sea buckthorn juice with Lactiplantibacillus plantarum

This work investigated the impact of malolactic fermentation on the metabolomic profile of sea buckthorn juice to optimize the fermentation process for flavor modification. Six strains of L. plantarum were used with varied pH of the juice, cell acclimation, and fermentation time. 1H-NOESY spectra were acquired from fresh and fermented juices with a total of 46 metabolites identified. Less sugars and quinic acid were metabolized at pH 2.7 while oxidation of ascorbic acid was reduced at pH 3.5. l-Malic acid, essential amino acids, and nucleosides were consumed early during fermentation while sugars in general were consumed later in the fermentation. If deacidification is the main target of fermentation, strains that produce less acids and ferment less sugars, shorter fermentation time, and lower starter pH should be used. Higher starter pH and longer fermentation time promote formation of antimicrobial compounds and potentially increase antioxidant stability. </p

Identification of the compositional changes in Orthosiphon stamineus leaves triggered by different drying techniques using 1H NMR metabolomics

BACKGROUND: Java tea is a well-known herbal infusion prepared from the leaves of Orthosiphon stamineus (OS). The biological properties of tea are in direct correlation with the primary and secondary metabolite composition, which in turn largely depends on the choice of drying method. Herein, the impact of three commonly used drying methods, i.e. shade, microwave and freeze drying, on the metabolite composition and antioxidant activity of OS leaves was investigated using proton nuclear magnetic resonance (1H NMR) spectroscopy combined with multivariate classification and regression analysis tools. RESULTS: A total of 31 constituents comprising primary and secondary metabolites belonging to the chemical classes of fatty acids, amino acids, sugars, terpenoids and phenolic compounds were identified. Shade-dried leaves were identified to possess the highest concentrations of bioactive secondary metabolites such as chlorogenic acid, caffeic acid, luteolin, orthosiphol and apigenin, followed by microwave-dried samples. Freeze-dried leaves had higher concentrations of choline, amino acids leucine, alanine and glutamine and sugars such as fructose and α-glucose, but contained the lowest levels of secondary metabolites. CONCLUSION: Metabolite profiling coupled with multivariate analysis identified shade drying as the best method to prepare OS leaves as Java tea or to include in traditional medicine preparation