A study on incorporation of giloy (Tinosporacordifolia) for the development of shelf-stable goat milk based functional beverage.

Goat milk has achieved significant place in human diet owing to its enormous therapeutic properties. There exists a scope of value-addition of goat milk to potentiate its health benefits by incorporating herbs and plants. Giloy (Tinospora cordifolia), a traditional medicinal plant with rich bioactive composition, can enhance the bioactive properties and shelf-life of goat milk. To this end, a study was conducted to develop shelf-stable giloy-goat milk beverage (GGB) by adding debittered giloy juice to goat milk (GM) and analyse the detailed product profile including proximate composition, bioactive properties, sensory, rheological, and structural characterisation. GGB resulted in two-fold increase (P < 0.05) in antioxidant activity and total phenolic content, thus enhancing the bioactive properties of the beverage as compared to GM. Further, increase in the particle size of GGB was observed along with components interaction, which was confirmed by FTIR, scanning electron and fluorescent microscopy. Storage stability studies indicated that bioactive properties of GGB remained unaffected (P > 0.05) by the sterilization process up to 90 days and sensory characteristics were not compromised till 105 days of storage. Therefore, the developed GGB is considered to be a shelf-stable beverage that retains its bioactive and sensory properties even after sterilization, making it a promising functional dairy product.

Dietary Guduchi (Tinospora cordifolia) enhanced the growth performance, antioxidative capacity, immune response and ameliorated stress-related markers induced by hypoxia stress in Nile tilapia (Oreochromis niloticus).

Nile tilapia can tolerate a wide range of farming conditions; however, fluctuations in the environmental conditions may impair their health status. The incorporation of medicinal herbs in aquafeed is suggested to overcome stressful conditions. In this study, dietary Guduchi (Tinospora cordifolia) was evaluated on the growth performance, antioxidative capacity, immune response, and resistance of Nile tilapia against hypoxia stress. Fish fed five diets incorporated with Guduchi at 0, 2, 4, 6, and 8 g/kg for 56 days then exposed with hypoxia stress for 72 h. The growth performance, feed intake, and feed efficiency ratio were significantly (P < 0.05) increased by including Guduchi in tilapia diets regardless of the inclusion level. Similarly, the lipase and protease activities were markedly (P < 0.05) increased in tilapia fed dietary Guduchi. The activities of lysozyme and bactericidal activities in serum and mucus, nitro-blue tetrazolium (NBT), and alternative complement activity (ACH50) were markedly (P < 0.05) enhanced in tilapia treated with Guduchi supplements regardless of the dose. Additionally, the activities of liver and intestinal superoxide dismutase, catalase, and glutathione peroxidase were markedly enhanced (P < 0.05) by including Guduchi in tilapia diets compared with the control. Before and after hypoxia stress, tilapia-fed dietary Guduchi had lower glucose and cortisol levels than fish-fed Guduchi-free diets (P < 0.05). In all groups, glucose and cortisol levels were markedly higher after hypoxia compared before hypoxia stress (P < 0.05). In conclusion, dietary Guduchi can be included at 5.17-5.49 g/kg to enhance the growth performance, digestive enzyme activity, immune and antioxidative responses, and the resistance of Nile tilapia against hypoxia stress.

Anti-dengue therapeutic potential of Tinospora cordifolia and its bioactives.

ETHNOPHARMACOLOGICAL RELEVANCE: Dengue is one of the most prevalent mosquito-borne viral infections. Moreover, due to the absence of appropriate curative and preventive measures against it, the mortality rate is increasing alarmingly. However, remarkable docking and clinical advances have been achieved with plant-based natural and conventional therapeutics. Tinospora cordifolia is one of the highly explored panaceas at the local level for its effective anti-dengue formulations. AIM OF THE STUDY: The present article aims for critical assessment of the data available on the anti-dengue therapeutic use of T. cordifolia. Efforts have also been made on the clinical and in-silico anti-dengue efficacy of this plant. The phytochemistry and the antiviral machinery of the plant are also emphasized. MATERIALS AND METHODS: The present article is the outcome of the literature survey on the anti-dengue effect of T. cordifolia. A literature survey was conducted from 2011 to 2024 using different databases with appropriate keywords. RESULTS: The present study confirms the anti-dengue potential of T. cordifolia. The plant can suppress the initiation of 'cytokine storm', vascular leakage and inhibition of various structural and NS proteins to exert its anti-dengue potential. Berberine and magnoflorine phytocompounds were highly explored for their anti-dengue potential. CONCLUSIONS: The present study concluded that T. cordifolia serves as an effective therapeutic agent for treating dengue. Further in-silico and clinical studies are needed so that stable, safe and efficacious anti-dengue drug can be developed. Besides, a precise antiviral mechanism of T. cordifolia against DENV infection is still needed.

Phytochemical analysis of Tinospora cordifolia and Withania somnifera and their therapeutic activities with special reference to COVID-19.

Various important medicines make use of secondary metabolites that are produced by plants. Medicinal plants, such as Withania somnifera and Tinospora cordifolia, are rich sources of chemically active compounds and are reported to have numerous therapeutic applications. The therapeutic use of medicinal plants is widely mentioned in Ayurveda and has folkloric importance in different parts of the world. The aim of this review is to summarize the phytochemical profiles, folkloric importance, and primary pharmacological activity of W. somnifera and T. cordifolia with emphasis on their action against the novel coronavirus.

Giloy: a potential anti-COVID-19 herb with propitious pharmacological attributes: a short review.

Plant-based medicine actually restores the balance in the body instead of treating the source of the disease. The strain of coronavirus (SAR-CoV-2) going to be more serious due to the lack of a reliable treatment option. Holistic treatment for this disease is in the form of Ayurveda as traditional medicine. As the infection of coronavirus is spreading like a wildfire, so the one way to fight is 'immunity'. Building immunity is the only way to stay safe and healthy and prepared themselves for the ongoing pandemic. In the current scenario, good immunity safeguard us from disease progression and prevention from this deadly virus. Giloy herb came into the limelight after the start of the COVID-19 pandemic due to its immunomodulatory and antiviral activity. The genome sequencing of Giloy is proved to be a breakthrough for controlling the COVID-19.Communicated by Ramaswamy H. Sarma.

Preparation, characterization, and safety assessment of statistical optimized probiotic supplemented herbal wine from Tinospora cordifolia.

Evidently proven medicinal benefits of Tinospora cordifolia and the growing demand of functional foods have created scientific interest in the functional beverage. Therefore, an attempt was made to prepare probiotic Lactiplantibacillus pentosus GSSK2 supplemented herbal wine having the benefits of both phytochemical and probiotic. Experimentally, fermentation of Tinospora cordifolia stem was found to be the most effective with ammonium dihydrogen phosphate, potassium phosphate, magnesium sulfate, isoleucine, and thiamine that yielded maximum ethanol (6.8 to 10%), total phenol (419 to 791.5 µg/ml), and antioxidants capacity (98.2 to 160.4 µmol/ml) after optimizing physical parameters, i.e., 20° Brix total soluble solid, pH 4.5, temperature 30 °C, and 10% (v/v) inoculum. Further, prepared herbal wine was supplemented separately with seven different probiotic strains and among these Lactiplantibacillus pentosus GSSK2 had the highest 88.6% survival rate compared with other probiotics and was safe showing 100% survivability of HEK-293 and THP-1 cells. Both herbal- and probiotic-supplemented herbal wine showed the antimicrobial potential against Gram-positive and Gram-negative bacteria as probiotic-supplemented herbal wine had 19-21 mm inhibition zone compared with 18-19 mm with herbal wine. LC-MS analysis of the probiotic-supplemented herbal wine revealed the presence of various phytochemicals such as alkaloids, diterpenoid lactone, glycoside, steroids having anti-bacterial, anti-oxidant, and anti-inflammatory potential. This is the first ever such study to demonstrate the antibacterial, antioxidant potential and safety of probiotic supplemented herbal wine in vitro.

Anti-F-Actin Antibody Positivity on Indirect Immunofluorescence Assay Following Chinese and Alternative Medicine Therapy: A Case Report.

Immunofluorescence on human epithelial type 2 cells is the standard screening assay for the detection of antinuclear antibodies (ANA). Cytoplasmic speckled patterns are a common finding. However, the less commonly reported ones include the cytoplasmic fibrillar patterns on indirect immunofluorescence technique (IIFT). The cytoplasmic fibrillar patterns include the cytoplasmic linear (AC-15), cytoplasmic filamentous (AC-16), and cytoplasmic segmental (AC-17). We report a case of cytoplasmic linear (F-actin) detected through IIFT during ANA screening in a 77-year-old man and later reconfirmed on liver mosaic biochip through IIFT on vascular smooth muscle substrate (VSM-47) without features suggestive of anti-smooth muscle antibody involvement post-complementary and alternative medicine therapy initiation.

Tinospora cordifolia (Guduchi/Giloy)-Induced Liver Injury: A Case Review.

Tinospora cordifolia (Guduchi/Giloy) is a relatively common herbal supplement whose use has recently become prominent in Southeast Asia. It was promoted to the public in India as an immunity booster, especially against the novel COVID-19. There have been reports, mostly from India, of an association between Guduchi/Giloy and liver injury. We present a 50-year-old female with a history of Hashimoto thyroiditis, who presented with abdominal discomfort and nausea of two weeks duration, which coincided with starting HistaEzeTM supplement containing Tinospora cordifolia. The vital signs upon presentation showed no significant abnormalities. Labs were significant for severely elevated transaminases; however, viral panels, autoimmune serologies, and imaging studies were unremarkable. Roussel Uclaf causality assessment method (RUCAM) score was at 6, which was indicative of probable drug/herb-induced liver injury. HistaEzeTM was discontinued, and the patient took a three-day course of oral steroids with significant interval improvement in clinical status, as evidenced by progressive normalization of the transaminases level. The transaminases decreased by greater than 50% within two weeks of discontinuation and trended back to baseline within three months. This case highlights the worldwide availability and use of Tinospora cordifolia, which can cause liver injury that appears to be idiosyncratic and possibly immune-mediated. Further research on the precise mechanism of its hepatotoxicity is warranted.

OECD-407 Driven 28-day-repeated-dose non-clinical safety evaluation of Tinospora cordifolia (Giloy) stem aqueous extract in Sprague-Dawley rats under GLP compliance.

Introduction: Tinospora cordifolia (Wild.) Hook.f. & Thomson (Giloy), has been widely used in the Ayurvedic system of medicine. However, some sporadic under-powered case studies have recently reported Tinospora cordifolia associated toxicity. Thus, following OECD 407 guidelines, a 28-day-repeated-dose-14-day-recovery toxicological evaluation of the aqueous extract of T. cordifolia stem (TCWE) was conducted under good laboratory practice (GLP), in Sprague-Dawley (SD) rats. Methods: 100, 300, and 1000 mg/kg/day of TCWE was given orally to designated treatment groups of either sex. Two separate 14-day recovery satellite groups received either vehicle control or 1000 mg/kg/day of TCWE. Results: In this study, TCWE was found safe up to a dose of 1000 mg/kg/day with no mortality or related toxicological manifestation in terms of clinical signs, ocular effects, hematology, urinalysis, clinical chemistry parameters, or macro- or microscopic changes in any organs. The satellite group did not show any adverse effect after 14-day recovery period. Thus, the No-Observed-Adverse-Effect-Level (NOAEL) of TCWE was determined to be 1000 mg/kg/day. Discussion: In conclusion, this study established the non-clinical safety of the aqueous extract of T. cordifolia stem, which confirms the age-old safe medicinal use of this herb, and also paves the path for future clinical research on formulations containing Tinospora cordifolia.

Herb-induced Liver Injury-A Guide to Approach. Lessons from the Tinospora cordifolia (Giloy) Case Series Story.

Background: Tinospora cordifolia (TC) is being increasingly consumed in India for its health and suggested immune-enhancing benefits in preventing and countering COVID-19. We previously published our experience of hepatotoxicity with self-medication of TC in six individuals. Since herb-induced liver injury (HILI) has been described with Tinospora crispa (TCR) consumption, it was contested that our patients may have mistakenly self-medicated with TCR which is similar in appearance to TC. Methods: We collected the four plant samples and two commercial preparations that were consumed by our patients for further analysis. The six samples underwent high performance thin layer chromatography phytochemical analysis and DNA barcoding studies for the confirmation of the genus and species. The four plant part samples which included stems and leaves were also analysed by a botanist for the characteristic morphological and microscopic features. Results: Based on morphological, microscopic, phytochemical and DNA studies, the four plant part samples were identified as TC. The two commercial preparations could not be analysed on phytochemical analysis or DNA barcoding studies due to other ingredients that most likely interfered with the analysis. The herb consumed by our study subjects was confirmed to be Tinospora cordifolia. Conclusion: We have highlighted the key morphological and phytochemical differences between these two species. We propose an algorithmic approach to accurately identify the implicated herb in cases of HILI. Future studies on causality need to focus on the serological/histopathological identification of active herb/metabolites in human tissues.

Unmasking the Many Faces of Giloy (Tinospora cordifolia L.): A Fresh Look on its Phytochemical and Medicinal Properties.

BACKGROUND: Plants have been used to treat health disorders and prevent diseases since antiquity in all civilizations including the Indian and Chinse. Giloy (Tinospora cordifolia L.) is one of the most important medicinal plants in Indian folk medicine and is commonly known as the "Heart-leaved Moonseed" or "Guduchi". Giloy is a perennial shrubby creeper of the Menispermaceae family. It is native to the tropical areas of India, Sri Lanka, Myanmar, Sothern Eastern Asia, Africa, and Australia. It attracted attention in the last few decades because its roots, stems, and leaves are used in folk medicine treating several disorders. METHODS: To reveal its potential, we reviewed the literature on this plant over the last five decades using several search engines. RESULTS: Giloy is recommended in diabetes, stomachache, jaundice, urinary problems, skin ailments, and prolonged diarrhea and dysentery. The medicinal properties of T. cordifolia extracts were attributed to its phytochemical content including steroids, alkaloids, diterpenoid lactones, and glycosides. CONCLUSION: In this review, we summarized the phytochemical content of T. cordifolia and its medicinal properties with special emphasis on its antioxidant, anti-inflammatory, and cytotoxic activities. We aimed to provide natural product researchers with a full picture of the potential applications of this plant in the development of food additives and nutraceuticals.

Potential Application of an Aqueous Extract of Tinospora Cordifolia (Thunb.) Miers (Giloy) in Oral Submucous Fibrosis-An In Vitro Study.

The in vitro antifibrotic activity of Tinospora cordifolia (Thunb.) Miers (giloy) was assessed to explore its potential for the management of oral submucous fibrosis. Epithelial cells dissociated from the tissue obtained from histopathologically normal oral mucosa during surgical extraction of third molars were cultured and fibrosis was induced by TGF-ß1 in the oral keratinocytes. Cell viability was assessed by MTT and comparative gene expression analysis was carried out in the fibrosis-induced oral keratinocytes treated with various concentrations of Tinospora cordifolia extract (TcE) for matricellular protein-related gene expression. Concentrations of 0.5 µg/mL and 1 µg/mL TcE demonstrated a significant reduction in the expression of CTGF, SERPINE1, COL1A1, FN1, MMP1, MMP2, MMP3, and TIMP2 and an increase in the expression of PLAU, COL3A1, TIMP1, and TIMP3. Although TcE was found to reduce the expression of several fibrotic genes and increase the expression of antifibrotic genes, a varied effect was found, causing increased expression of COL3A1 and decreased expression of TIMP2 on TGF-ß1-induced human buccal epithelial cells. However, further studies are warranted to assess the exact mechanism of antifibrotic activity and its clinical applications.

Prospective, Randomized, Open-Label, Blinded End Point, Two-Arm, Comparative Clinical Study to Evaluate the Efficacy and Safety of a Fixed Ayurvedic Regimen (FAR) as Add-on to Conventional Treatment in the Management of Mild and Moderate COVID-19 Patients.

BACKGROUND: The traditional healthcare systems are being avidly looked into in the quest for effective remedies to tackle the menace of COVID-19 pandemic. OBJECTIVE: This was a prospective randomized, controlled open-label, blinded end point (PROBE) study to evaluate the efficacy and safety of a fixed ayurvedic regimen (FAR) as an add-on to conventional treatment/standard of care (SOC) in the management of mild-to-moderate COVID-19 infection. METHODOLOGY: A total of 68 patients were recruited who consumed either FAR + SOC (n = 35) or SOC only (n = 33) for 28 days. Primary outcomes assessed were mean time required for clinical recovery and proportion of patients showing clinical recovery between the groups. Secondary outcomes assessed included mean time required for testing SARS-CoV-2 negative, change in clinical status on World Health Organization (WHO) ordinal scale, number of days of hospitalization, change in disease progression and requirement of oxygen/intensive care unit admission/ventilator support/rescue medication, health status on WHO quality of life (QOL) BREF and safety on the basis of occurrence of adverse event/serious adverse event (AE/SAE) and changes in laboratory parameters. RESULTS: Patients consuming FAR as an add-on SOC showed faster clinical recovery from the day of onset of symptoms by 51.34% (P < 0.05) as compared to SOC group. A higher proportion of patients taking FAR recovered within the first 2 weeks compared to those taking only SOC. It was observed that 5 times more patients recovered within 7 days in FAR group when compared to SOC (P < 0.05) group. An earlier clinical recovery was observed in clinical symptoms such as sore throat, cough, loss of taste and myalgia (P < 0.05). Improvement in postclinical symptoms such as appetite, digestion, stress and anxiety was also obs served to be better with the use of FAR. Requirement of rescue medications such as antipyretics, analgesics and antibiotics was also found to be reduced in the FAR group (P < 0.05). FAR showed a significant improvement in all the assessed domains of QOL. None of the AEs/SAE reported in the study were assessed to be related to the study drugs. Further, FAR did not produce any significant change in the laboratory safety parameters and was assessed to be safe. CONCLUSION: FAR could be an effective and safe add-on ayurvedic regimen to standard of care in the management of mild and moderate COVID-19 patients. CTRI number: CTRI/2020/09/027914.

Giloy Ghanvati (Tinospora cordifolia (Willd.) Hook. f. and Thomson) Reversed SARS-CoV-2 Viral Spike-Protein Induced Disease Phenotype in the Xenotransplant Model of Humanized Zebrafish.

The current Severe Acute Respiratory Syndrome disease caused by Coronavirus-2 (SARS-CoV-2) has been a serious strain on the healthcare infrastructure mainly due to the lack of a reliable treatment option. Alternate therapies aimed at symptomatic relief are currently prescribed along with artificial ventilation to relieve distress. Traditional medicine in the form of Ayurveda has been used since ancient times as a holistic treatment option rather than targeted therapy. The practice of Ayurveda has several potent herbal alternatives for chronic cough, inflammation, and respiratory distress which are often seen in the SARS-CoV-2 infection. In this study we have used the aqueous extracts of Tinospora cordifolia (willd.) Hook. f. and Thomson in the form of Giloy Ghanvati, as a means of treatment to the SARS-CoV-2 spike-protein induced disease phenotype in a humanized zebrafish model. The introduction of spike-protein in the swim bladder transplanted with human lung epithelial cells (A549), caused an infiltration of pro-inflammatory immune cells such as granulocytes and macrophages into the swim bladder. There was also an increased systemic damage as exemplified by renal tissue damage and increased behavioral fever in the disease induction group. These features were reversed in the treatment group, fed with three different dosages of Giloy Ghanvati. The resultant changes in the disease phenotype were comparable to the group that were given the reference compound, Dexamethasone. These findings correlated well with various phyto-compounds detected in the Giloy Ghanvati and their reported roles in the viral disease phenotype amelioration.

Tinospora cordifolia (Thunb.) Miers (Giloy) inhibits oral cancer cells in a dose-dependent manner by inducing apoptosis and attenuating epithelial-mesenchymal transition.

BACKGROUND: Tinospora cordifolia (Thunb.) Miers (Giloy) has been applied successfully as an anti-inflammatory, anti-diabetic, and even as an anti-cancer agent. Yet, to date, the application of Giloy has not been explored concerning oral cancer. OBJECTIVES: To assess the effect of T cordifolia (Thunb.) Miers (Giloy) extract (TcE) on an oral cancer cell line. METHODS: AW13516 (oral cancer cell line) cells were treated with the prepared aqueous extract of TcE for 24 h at various concentrations ranging between 5 µg/ml and 100 µg/ml and compared with control (cells without treatment). Thee effect of the extracts on apoptosis was assessed by through Annexin V flow cytometry assay and Luminometry based assessment of Caspase 8, 9 and caspase 3/7 activity. RNA was isolated from treated cells and gene expression of selected metastatic genes (MMP1, MMP10, and CXCL8); epithelial-mesenchymal stem cell genes (TWIST1, SNAIL, ZEB1, Oct4) and stemness related genses (Nanog, Sox2) were analyzed by using a quantitative real-time PCR system. The experiments were performed in triplicates. RESULTS: Aqueous extract of TcE was found to induce apoptosis inducer in AW13516 cells in a concentration-dependent manner and was potent even at a low concentration of 5 µg/ml. The apoptosis induction was confirmed with the caspase activity assay. Treatment of the cells with the extract for 24 h exhibited a significant decrease in the expression of EMT genes in a dose-dependent manner without an effect on the metastatic genes. CONCLUSION: Aqueous extract of TcE induces apoptosis-mediated cell death in the oral cancer cell line AW13516 while attenuating its potential for epithelial mesenchymal transition.

Comparative retrospective open-label study of ayurvedic medicines and their combination with allopathic drugs on asymptomatic and mildly-symptomatic COVID-19 patients.

INTRODUCTION: Treatment for COVID-19 was ambiguous in the beginning of the pandemic. At that time, the conventional medical system was grappling to cope with the rapidly spreading pandemic. The potential of Ayurveda, one of the branches of traditional Indian medicine (TIM), with a 5000 year old history, employing medicines derived from plants and other natural sources, against COVID-19 has been explored through a comparative retrospective open-label study. METHODS: Reported here are the remedial effects of Ayurvedic medicines alone or in combination with Allopathic treatment on 59 asymptomatic or mildly symptomatic COVID-19 patients, across multiple COVID-19 care centers in Ahmedabad, India. The patients were confirmed for COVID-19 infection through RT-qPCR of nasopharyngeal swabs. With informed consents from the patients, the sourced data was divided into 'Allopathic and Ayurvedic' [AlloAyur] (n = 41) and 'Ayurvedic only'[Ayur] (n = 18) groups, based on the type of treatment the patients decided to receive, that is Ayurvedic medicines with Allopathic treatment or Ayurvedic medicines alone, respectively. Ayurvedic medicines included oral doses and nasal drops; the dosage and regime were decided based on the recommendations from Ayurvedic texts. The Allopathic medicines included Azithromycin, Vitamin-C and anti-histamines. Acetaminophen was also administered when necessary, by the attending physician. The patients were observed for symptomatic improvement. RESULTS: Primary outcome of this study was the symptomatic relief from COVID-19. Data collected over a period of two months, showed that more patients exhibited symptomatic relief in Ayur goup (83.33 %) than in the AlloAyur group (48.78 %) within the first 13 days of treatment. No visible adverse effects were observed. This indicated faster and safe symptomatic resolution among those treated with Ayurvedic medicines alone. CONCLUSION: Patients receiving only Ayurvedic medicines on average were symptomatically relieved faster than those receiving Allopathic and Ayurvedic medicines together.

Herbal Immune Booster-Induced Liver Injury in the COVID-19 Pandemic - A Case Series.

The COVID-19 pandemic has resulted in widespread use of complementary and alternative medicines. Tinospora cordifolia is a widely grown shrub which has been commonly used in India's traditional system of Ayurveda for its immune booster properties and has been extensively used as prophylaxis against COVID-19. Six patients (4 women, 2 men) with a median (IQR) age of 55 years (45-56) and with an history of Tinospora cordifolia consumption presented with symptoms of acute hepatitis during the study period of 4 months in the COVID-19 pandemic. The median (IQR) duration of Tinospora cordifolia consumption was 90 days (21-210). The median (IQR) peak bilirubin and AST were 17.5 mg/dl (12.2-24.9) and 1350 IU/ml (1099-1773), respectively. The patients had either a definite (n = 4) or probable (n = 2) revised autoimmune hepatitis score with an autoimmune pattern of drug-induced liver injury on biopsy. Four of these patients (all women) had underlying silent chronic liver disease of possible autoimmune etiology associated with other autoimmune diseases - hypothyroidism and type 2 diabetes mellitus. One of the three patients treated with steroids decompensated on steroid tapering. The other five patients had resolution of symptoms, liver profile, and autoimmune serological markers on drug withdrawal/continuing steroid treatment. The median (IQR) time to resolution from discontinuing the herb was 86.5 days (53-111). Tinospora cordifolia consumption seems to induce an autoimmune-like hepatitis or unmask an underlying autoimmune chronic liver disease, which may support its immune stimulant mechanism. However, the same mechanism can cause significant liver toxicity, and we recommend that caution be exercised in the use of this herb, especially in those predisposed to autoimmune disorders. Besides, in patients presenting with acute hepatitis, even in the presence of autoimmune markers, a detailed complementary and alternative medicine history needs to be elicited.

UPLC-DAD Assisted Phytochemical Quantitation Reveals a Sex, Ploidy and Ecogeography Specificity in the Expression Levels of Selected Secondary Metabolites in Medicinal Tinospora cordifolia: Implications for Elites' Identification Program.

BACKGROUND: Medicinal phytochemistry involving UPLC-DAD in an exhaustive analysis involving quantification of eight commercially important phytochemicals viz. syringin, cordifolioside A, magnoflorine, tinocordiside, palmatine, 20ß-hydroxyecdysone, L-tetrahydropalmatine and berberine has been done in 143 accessions from eight states and the union territories of Delhi and Jammu & Kashmir of India representing three different ploidy levels viz. diploid (2x), triploid (3x) and synthetic tetraploid (4x). The study was done to assess the effect of sex, ploidy level and ecogeography on the expression level of secondary metabolites in stems of dioecious, medicinally important shrub Tinospora cordifolia. METHODS: Two different UPLC-DAD methods were used for the quantification of eight selected phytochemicals from the alcoholic stem extracts of T. cordifolia accessions. The Waters Acquity UPLC system hyphenated to the QTOF micromass system, equipped with PDA and ESI-Q-TOF detectors was utilized for the quantitative analysis, Mass Lynx v 4.0 software was used for data analysis. RESULTS: Significant quantitative changes were observed in the analysed secondary metabolites among different accessions of T. cordifolia. The triploid (3x) cytotypes revealed higher amounts of seven out of eight analysed secondary metabolites than diploids and only 20ß-hydroxyecdysone was observed to be present in significantly higher amount in diploid cytotypes. Further, at the tetraploid level, novel induced colchiploid (synthetic 4x) genotypes revealed increase in the yield of all of the analysed eight phytochemicals than their respective diploid counterparts. The quantity of active principles in tetraploid cytotypes were also higher than the average triploid levels at multiple locations in five out of eight tested phytochemicals, indicating the influence of ploidy on expression levels of secondary metabolites in T. cordifolia. Additionally, at each of the three ploidy levels (2x, 3x and synthetic 4x), a significant sex specificity could be observed in the expression levels of active principles, with female sex outperforming the male in the content of some phytochemicals, while others getting overexpressed in the male sex. The manifestation of diverse ecogeographies on secondary metabolism was observed in the form of identification of high yielding accessions from the states of Madhya Pradesh, Delhi and Himachal Pradesh and the Union territory of Jammu & Kashmir. Two triploid female accessions that contained approximately two- to eight fold higher amounts of five out of the eight analysed phytochemicals have been identified as superior elites from the wild from the states of Delhi and Madhya Pradesh. CONCLUSION: The paper shows the first observations of ploidy specificity along with subtle sex and ecogeography influence on the expression levels of secondary metabolome in T. cordifolia.