Pomegranate Juice Enhances Iron Dialysability and Assimilation in In-Vitro Cell Free and Cell-Based Models.

Despite concerted programmatic efforts iron deficiency anemia (IDA) continues to be a global health problem. Per Ayurveda, an Indian Traditional Medical System, natural plant materials such as pomegranate juice (PJ) is indicated as a food supplement to manage IDA. We hypothesized that pomegranate could play a role in improving absorption and assimilation of iron. A cell free in vitro model simulating the gastric and intestinal digestive processes coupled with cell based (Caco-2 and HepG2) models were used to assess iron (FeSO4 form) dialysability in the presence of PJ. Iron assimilation into cells was measured in terms of the cellular ferritin content. PJ (containing ~13 mg/100 ml natural ascorbic acid equivalent) increased the dialysability of iron by >3 fold when compared to control in the cell free model. An equivalent concentration of ascorbic acid alone increased it only by 1.6-fold. PJ increased the iron uptake in Caco2 cells by ~6-fold and ferritin content by 30% when compared to the ascorbic acid control. Similarly, PJ enhanced the iron uptake in HepG2 cells by ~3 fold and iron assimilation by about 50%. This study establishes a scientific evidence for Ayurveda's claim of using pomegranate in the management of IDA by facilitating iron absorption and assimilation. It provides a simple solution for addressing the global problem of IDA. Synergistic action of multiple phytochemicals, over and above ascorbic acid, in PJ may be responsible for improving iron bioavailability.

Can Guduchi (Tinospora cordifolia), a well-known ayurvedic hepato-protectant cause liver damage?

Ayurveda is a centuries old traditional medicine practiced in India even today. There are certain safe medicinal plants with well-established medicinal properties both in clinical practice as well as in modern scientific publications. Guduchi or Tinospora cordifolia (Willd.) Miers (Menispermaceae), is one such medicinal plant that has well known anti-inflammatory, immune-modulatory and other safe therapeutic applications including hepato-protection, because of which it was recommended by the Ministry of AYUSH, Government of India to be used in COVID-19 care. Therefore, Aabha Nagral's article "Herbal Immune Booster-Induced Liver Injury in the COVID-19 Pandemic-a Case Series," published in 2021, was unanticipated. The article recounted histologically documented clinical cases of six patients who developed drug-induced autoimmune-like hepatitis after reported consumption of Guduchi or Guduchi containing formulations during the COVID-19 pandemic. Since the Ayurveda practitioners vouch by the safety of T. cordifolia (TC), it was felt that the story needed to be further scrutinized. This article reviews the botanical entities, the substitutes and adulterants of species used as Guduchi, their pharmacological and toxicological properties. While the authentic botanical entity of Guduchi is TC, Tinospora sinensis and Tinospora crispa are also commonly traded in the Indian subcontinent as Guduchi or Giloy. Among these species, T. crispa is known to induce heapto-toxicity. In Nagral's article, there were variations in the reported six cases in terms of patient history and TC/TC product consumption. More importantly, the botanical authenticity of the consumed products was not investigated. A review of published literature indicates that it is unlikely that the authentic TC could have induced autoimmune-like hepatitis of the patients. It is probable that a wrong species was self-administered by the patients. It is worth following up with the cases (patients), to investigate details of the products, so that other consumers do not suffer. Nagral's article however does highlight the serious issue of adulteration in herbal markets and the need for establishing a robust pharmacovigilant system in India.

Storing drinking-water in copper pots kills contaminating diarrhoeagenic bacteria.

Microbially-unsafe water is still a major concern in most developing countries. Although many water-purification methods exist, these are expensive and beyond the reach of many people, especially in rural areas. Ayurveda recommends the use of copper for storing drinking-water. Therefore, the objective of this study was to evaluate the effect of copper pot on microbially-contaminated drinking-water. The antibacterial effect of copper pot against important diarrhoeagenic bacteria, including Vibrio cholerae O1, Shigella flexneri 2a, enterotoxigenic Escherichia coli, enteropathogenic E. coli, Salmonella enterica Typhi, and Salmonella Paratyphi is reported. When drinking-water (pH 7.83 +/- 0.4; source: ground) was contaminated with 500 CFU/mL of the above bacteria and stored in copper pots for 16 hours at room temperature, no bacteria could be recovered on the culture medium. Recovery failed even after resuscitation in enrichment broth, followed by plating on selective media, indicating loss of culturability. This is the first report on the effect of copper on S. flexneri 2a, enteropathogenic E. coli, and Salmonella Paratyphi. After 16 hours, there was a slight increase in the pH of water from 7.83 to 7.93 in the copper pots while the other physicochemical parameters remained unchanged. Copper content (177 +/- 16 ppb) in water stored in copper pots was well within the permissible limits of the World Health Organization. Copper holds promise as a point-of-use solution for microbial purification of drinking-water, especially in developing countries.

University Wellness Program-A Pedagogic Innovation to Nudge Wellness and Sustainability Among Students.

Anthropocentric activities have induced climate change, threatened planetary health, and harmed human health and wellness. The changing lifestyles, dietary patterns and digital obsession have affected the mental and physical health, particularly of the youth. University campuses reflect the challenges faced by the society at large and therefore make for an ideal ecosystem to initiate positive changes toward wellness and sustainability. The energy of ~200 million university students globally is largely unleveraged for facing these challenges. Values of empathy and sustainable living are crucial to be inculcated, alongside technical and managerial skills for leading the mass transformation. This article describes a novel pedagogic approach called the University Wellness Program (UWP). The aim of UWP is to equip students with technical and leadership skills to achieve wellness and campus sustainability. That is, UWP is a platform that facilitates the students to design and implement multi-disciplinary projects that address campus related challenges. In the process, they acquire the necessary soft and technical skills to solve real-life problems. The durability of UWP is secured since the projects and activities are explicitly linked to existing curricula and evaluation system of the university. The strategy and framework adopted, and the early experiences of implementing UWP are shared. UWP is amenable for replication globally and has the potential to create change-makers.

De novo genome assembly and annotation of gall-forming medicinal plant Pistacia chinensis subsp. integerrima (J. L. Stewart ex Brandis) Rech. f.

Pistacia chinensis subsp. integerrima is one of the medicinal plants, well known for gall formation and popularly used in Ayurveda to treat various systemic diseases such as chronic disorders, respiratory problems, etc. P. integerrima genome characterization will aid in the study of Pistacia genes and pathways involved in therapeutic application. To understand the biological characteristics of this plant and to gain the genetic insight into the biosynthesis of its natural compounds, the whole genome of P. integerrima and its leaf transcriptome was sequenced using Illumina sequencing technology. The sequenced genome was functionally annotated, and gene prediction was performed with integrated genome annotation workflow. The pathway analysis was carried out using KEGG database. We obtained a draft genome assembly of 462 Mb with N50 16,145 bp. A total of 39,452 genes were found, and 18,492 of these contained RNA or protein evidence. We characterized the genes involved in biosynthetic pathways of different plant secondary metabolites such as flavonoids and terpenoids. Also, we identified miR397 and miR828 family noncoding RNA; which mainly targets the laccase (LCA) and MYB protein functioning respectively. Phylogeneic analysis showed that P. integerrima is genetically more closer to P. vera. In this study, we attempt to explore the whole genome information of P. integerrima which will provide a genomic insight in the future for omics studies as well as serves as valuable resource for the molecular characterization of medicinal compounds.

Development of Randomly Amplified Polymorphic DNA Based SCAR Marker for Identification of Ipomoea mauritiana Jacq (Convolvulaceae).

Vidari is an Ayurvedic herbal drug used as aphrodisiac, galactagogue and is also used in the preparation of Chyavanaprash. Tubers of Ipomoea mauritiana Jacq. (Convolvulaceae), Pueraria tuberosa (Roxb. ex Willd.) DC (Fabaceae), Adenia hondala (Gaertn.) de Wilde (Passifloraceae) and pith of Cycas circinalis L. (Cycadaceae) are all traded in the name of Vidari, creating issues of botanical authenticity of the Ayurvedic raw drug. DNA-based markers have been developed to distinguish I. mauritiana from the other Vidari candidates. A putative 600-bp polymorphic sequence, specific to I. mauritiana was identified using randomly amplified polymorphic DNA (RAPD) technique. Furthermore, sequence characterized amplified region (SCAR) primers (IM1F and IM1R) were designed from the unique RAPD amplicon. The SCAR primers produced a specific 323-bp amplicon in authentic I. mauritiana and not in the allied species.

Nuclear ribosomal DNA - ITS region based molecular marker to distinguish Gmelina arborea Roxb. Ex Sm. from its substitutes and adulterants.

Roots of Gmelina arborea (Gambhari) is a medicinally important raw drug traded in India. However, Gmelinaasiatica and Mallotus nudiflorus are also found in the raw drug markets as Gambhari. The current study aims to identify molecular markers based on the nuclear ribosomal DNA - ITS1 region to distinguish the authentic species from substitute/adulterants. The nuclear ribosomal internal transcribed spacer 1 (ITS1) was amplified to identify species-specific markers using universal primers. Based on the sequence of the ITS region, specific primers were designed for G. arborea, G. asiatica and M. nudiflorus which efficiently amplified 142 bp, 93 bp and 150 bp of the ITS1 region of the respective species. The notable feature of this molecular method is that it is technically accurate, practically convenient and suitable for analyzing large numbers of samples. This study demonstrates that the ITS1 region can be used for reliable authentication of medicinal plants and detection of adulterants and substitutes of Gambhari.

Sugarcane Molasses - A Potential Dietary Supplement in the Management of Iron Deficiency Anemia.

Iron deficiency anemia (IDA) is a serious public health problem that debilitates ∼1.6 billion people globally every year, the majority being pregnant women and children from developing countries. In India, for example, in spite of several operational programs at the national level using iron-folic acid and other allopathic interventions, IDA is still prevalent. Traditional medicines, such as Ayurveda, prescribe herbal formulations containing sugarcane derivatives for the management of pandu, a condition similar to IDA. This article reviews molasses, a sugar industry by-product, as a potential raw material to develop nutraceutical products for IDA. Molasses contains iron and its absorption enhancers, such as sulfur, fructose, and copper, which make it a potential dietary supplement for IDA. More research, product development, and evidence of safety and efficacy of molasses in IDA management can provide a tasty and cost-effective dietary supplement, particularly for children. However, there are challenges, such as competition for raw material from refined sugar manufacturers, quality control, etc., that need to be overcome.

A prospective comparative field study to evaluate the efficacy of a traditional plant-based malaria prophylaxis.

BACKGROUND: An unceasing threat of resistance of malarial parasites to available antimalarial drugs makes the development of new drugs imperative. Natural plant-based products are an alternative source for discovering new antimalarial drugs. AIM: To determine the prophylactic efficacy of a traditionally used plant-based drug on prevention of malaria in endemic villages of Odisha, India. METHODS: A total of 267 healthy human volunteers of both sexes, aged 18-60 years were enrolled in Odisha, India, to receive either minimum 20 doses of aqueous extract of Traditional Plant-based Malaria Prophylactic drug 74, twice a week (experimental group), or no drug (control group) for 14 weeks. The primary criterion was the occurrence of malaria positive cases confirmed through expert microscopy during the study period. Analyses were by per-protocol (PP) and modified intention-to-treat (mITT). RESULTS: A significant (P < 0.01) reduction (64%) of malaria incidence was observed in the experimental group compared to control group, 12.3% and 26.6%, respectively, as PP analysis. However, the reduction was nonsignificant as per mITT analysis (P = 0.22). The experimental group showed a relative risk of 0.36 compared to control group. CONCLUSION: This preliminary study constitutes a potential "proof of concept" for the development of malaria prophylactic drug and provide a scientific basis for the use of traditional remedy as a malaria preventive by tribal populations in India.

Exploring Ayurvedic Knowledge on Food and Health for Providing Innovative Solutions to Contemporary Healthcare.

Ayurveda, a traditional system of medicine that originated over three millennia ago in the South Asian region, offers extensive insights about food and health based on certain unique conceptual as well as theoretical positions. Health is defined as a state of equilibrium with one's self (svasthya) but which is inextricably linked to the environment. Ayurvedic principles, such as the tridosa (three humors) theory, provide the relationship between the microcosm and the macrocosm that can be applied in day-to-day practice. Classical Ayurveda texts cover an array of themes on food ranging from diversity of natural sources, their properties in relation to seasons and places and to their specific function both in physiological and pathological states. The epistemic perspective on health and nutrition in Ayurveda is very different from that of biomedicine and modern nutrition. However, contemporary knowledge is reinventing and advancing several of these concepts in an era of systems biology, personalized medicine, and the broader context of a more holistic transition in sciences in general. Trans-disciplinary research could be important not only for pushing the boundaries of food and health sciences but also for providing practical solutions for contemporary health conditions. This article briefly reviews the parallels in Ayurveda and biomedicine and draws attention to the need for a deeper engagement with traditional knowledge systems, such as Ayurveda. It points out that recreation of the methodologies that enabled the holistic view point about health in Ayurveda may unravel some of the complex connections with Nature.

Abhava pratinidhi dravya: A comparative phytochemistry of Ativisha, Musta and related species.

Authentic Ativisha (Aconitum heterophyllum) is a rare, endangered Himalayan species. Ayurveda classical texts of c. 15(th)-16(th) century, introduced "abhava-pratinidhi dravya" concept, wherein Ativisha was categorized as an abhava dravya (unavailable drug) and Musta (Cyperus rotundus) was suggested as a pratinidhi dravya (substitute) for it. C. rotundus is a weed, abundantly available pan-India. Cryptocoryne spiralis (Naattu Athividayam) and Cyperus scariosus (Nagarmotha) are also traded as Ativisha and Musta, respectively. Yet, there are no scientific studies to validate the use of substitutes. A. heterophyllum bears no similarity in terms of botanical classification with the other candidates. This article reviews published literature with an emphasis to look for similar phytochemicals or groups of phytochemicals in the species that could contribute to similar pharmacological activities, thereby supporting the drug substitution from a bio-medical perspective. Alkaloids like atisine were found to be the main focus of studies on A. heterophyllum, whereas for the Cyperus spp., it was terpenoids like cyperene. Although alkaloids and terpenoids were reported from both species, alkaloids in C. rotundus and terpenoids in A. heterophyllum were minor constituents. Reports on phytochemicals on Cryptocoryne spiralis and C. scariosus were very limited. Despite no significant similarities in chemical profiles reported, the dravyaguna (Ayurvedic drug classification) of Ativisha and Musta was quite similar warranting further exploration into the bio-functional aspects of the drug materials.

Pharmacology of Ativisha, Musta and their substitutes.

The Ayurvedic literature during the medieval period suggests the use of Musta (Cyperus rotundus), a common weed, as a pratinidhi dravya (substitute) for Ativisha (Aconitum heterophyllum), an endangered species. Contemporary Ayurvedic practice also uses Cryptocoryne spiralis, (known as Naattu Atividayam in South India) and Nagaramusta (Cyperus scariosus) as substitutes for Ativisha and Musta, respectively. This article reviews published literature on the pharmacology of the above four species. Both A. heterophyllum and C. rotundus are reported to possess antiinflammatory, antipyretic, antibacterial and antidiarrhoeal properties, while antiinflammatory and antibacterial activities are attributed to C. scariosus. No reports exist on the bioactivity of Cryptocoryne spiralis. It is interesting to note that other than the veerya which is different, the biological properties of Ativisha and Musta are similar according to Ayurvedic classification of dravyaguna. This is also supported by modern pharmacological studies, which show that, both A. heterophyllum and C. rotundus have antidiarrheal, antipyretic, antiinflammatory, antihyperlipidemic and hypoglycemic activities. However, the similarities between the discussed species cannot be attributed to their phytochemical composition or taxonomical classification as these are quite distinct. The dravyaguna method of classifying materials, which we are calling as "pharmaco-taxonomy", offers a unique way of classifying those plant materials which lack similarity at the botanical or chemical level, but are similar at the level of biological functions.

Proposed correlation of modern processing principles for Ayurvedic herbal drug manufacturing: A systematic review.

Quality Ayurvedic herbal medicines are potential, low-cost solutions for addressing contemporary healthcare needs of both Indian and global community. Correlating Ayurvedic herbal preparations with modern processing principles (MPPs) can help develop new and use appropriate technology for scaling up production of the medicines, which is necessary to meet the growing demand. Understanding the fundamental Ayurvedic principles behind formulation and processing is also important for improving the dosage forms. Even though Ayurvedic industry has adopted technologies from food, chemical and pharmaceutical industries, there is no systematic study to correlate the traditional and modern processing methods. This study is an attempt to provide a possible correlation between the Ayurvedic processing methods and MPPs. A systematic literature review was performed to identify the Ayurvedic processing methods by collecting information from English editions of classical Ayurveda texts on medicine preparation methods. Correlation between traditional and MPPs was done based on the techniques used in Ayurvedic drug processing. It was observed that in Ayurvedic medicine preparations there were two major types of processes, namely extraction, and separation. Extraction uses membrane rupturing and solute diffusion principles, while separation uses volatility, adsorption, and size-exclusion principles. The study provides systematic documentation of methods used in Ayurveda for herbal drug preparation along with its interpretation in terms of MPPs. This is the first step which can enable improving or replacing traditional techniques. New technologies or use of existing technologies can be used to improve the dosage forms and scaling up while maintaining the Ayurvedic principles similar to traditional techniques.

nrDNA ITS sequence based SCAR marker to authenticate Aconitum heterophyllum and Cyperus rotundus in Ayurvedic raw drug source and prepared herbal products.

To authenticate Ayurvedic medicinal plants Ativisha (Aconitum heterophyllum) and Musta (Cyperus rotundus) at the raw drug source and in prepared herbal products, nrDNA ITS sequence based SCAR markers were designed and validated spp.-specific SCAR primers gave amplicon of 415 bp and 134 bp, respectively, in authentic species. The SCAR primers (Cyr-FP and Cyr-RP) could identify tissue sample containing 750 µg to 4.76 mg/100mg of Musta in complex mixtures of DNA extracted from commercial herbal drugs. Ativisha could not be identified through SCAR markers suggesting that authentic species may not been used to prepare herbal drugs despite its being labelled as one of the ingredients in formulations. Analysis of individual tubers of Ativisha and Musta assures the presence of admixtures in raw drug trade of Ativisha, indicates the need to monitor the basic raw material supply and concludes, supplying plant materials through cultivation to manufacturing industries can minimize the risks of adulteration.

Botanical identity of plant sources of Dasamula drugs through an analysis of published literature.

BACKGROUND: Dasamula (DM) is a top-traded group of medicinal plants used by the Ayurvedic industry. Through literature survey and analysis, this article has enlisted the botanical sources of DM, as correlated by several scholars. Such a list is not available from any single, earlier publication. It brings to light the confusion that exists in terms of botanical sources correlated to Ayurvedic entities. There is quite a bit of difference in the botanical correlation, parts, and substitutes reported in the different scholarly works, particularly for Prsniparni, and Agnimantha. For e.g., is Uraria picta the original intended Prsniparni, as the Ayurvedic Formulary of India (AFI) stipulates or is it U. lagopoidiodes or Desmodium gangeticum as other scholars report? While AFI provides two botanical correlations to Agnimantha in its two editions, namely Premna integrifolia and Clerodendrum phlomidis, other scholars correlate it to other Premna and Clerodendrum species. Why has AFI provided stem bark and whole plant as substitutes for roots of DM? Are substitutes recommended by AFI only for ecological or practical convenience or is there an Ayurvedic or pharmacological explanation for the same? AIM: There are many species used in the name of Dasamula,, in this article all the species are listed out to find the differences in the usage of the drugs. MATERIALS AND METHODS: Ayurveda texts and lexicons along with the texts which have done correlation work were considered to arrive at a list of various species used as Dasmula. RESULTS AND CONCLUSION: Since neither the methodology nor the logic behind the correlation have been discussed in these scholarly works, including the AFI, the same is not available for analysis or scrutiny. Such a list as provided in this article can form an essential base for a much needed systematic approach at etymological analysis, botanical correlation, and further scientific work to establish legitimacy of substitutes prescribed.

Development of ITS sequence-based markers to distinguish Berberis aristata DC. from B. lycium Royle and B. asiatica Roxb.

The stems of Berberis aristata DC. (Berberidaceae) are used in the South Asian traditional medicine as a key ingredient in formulations for eye care, skin diseases, jaundice, rheumatism and also in diabetes. B. lycium Royle and B. asiatica Roxb. are traded as equivalents of B. aristata. Conventional macro-morphology and microscopic examination does not aid in critically distinguishing the three species. DNA markers were developed by amplifying and sequencing the complete internal transcribed spacer region (ITS1, 5.8S rRNA and ITS2) from the genomic DNA, using universal primers. The markers developed are efficient and reliable in authenticating B. aristata, B. asiatica and B. lycium. These are useful as molecular pharmacognostic tool in quality control of raw drugs.

Plant-based Rasayana drugs from Ayurveda.

Rasayana tantra is one of the eight specialties of Ayurveda. It is a specialized practice in the form of rejuvenative recipes, dietary regimen, special health promoting behaviour and drugs. Properly administered Rasayana can bestow the human being with several benefits like longevity, memory, intelligence, freedom from diseases, youthful age, excellence of luster, complexion and voice, optimum strength of physique and sense organs, respectability and brilliance. Various types of plant based Rasayana recipes are mentioned in Ayurveda. Review of the current literature available on Rasayanas indicates that anti-oxidant and immunomodulation are the most studied activities of the Rasayana drugs. Querying in Pubmed database on Rasayanas reveals that single plants as well as poly herbal formulations have been researched on. This article reviews the basics of Rasayana therapy and the published research on different Rasayana drugs for specific health conditions. It also provides the possible directions for future research.

Cyperus rotundus, a substitute for Aconitum heterophyllum: Studies on the Ayurvedic concept of Abhava Pratinidhi Dravya (drug substitution).

In the absence of a desired first choice medicinal herb, classical Ayurveda recommends use of a functionally similar substitute. Post 16th century Ayurvedic texts and lexicons give specific examples of possible substitutes. Here we report a preliminary study of one such Ayurvedic substitution pair: Musta (Cyperus rotundus L., Cyperaceae), a common weed, for the rare Himalayan species, Ativisha (Aconitum heterophyllum Wall. ex Royle; Ranunculaceae). The study's strategy was to use modern phytochemical and pharmacological methods to test the two herbs for biochemical and metabolic similarities and differences, and literary studies to compare their Ayurvedic properties, a novel trans-disciplinary approach. No previous scientific paper has compared the two herbs' bioactivities or chemical profiles. Despite being taxonomically unrelated, the first choice, but relatively unavailable (Abhava) plant, A. heterophyllum, and its substitute (Pratinidhi) C. rotundus, are not only similar in Ayurvedic pharmacology (Dravyaguna) profile, but also in phytochemical and anti-diarrheal properties. These observations indicate that Ayurveda may attach more importance to pharmacological properties of raw drugs than to their botanical classification. Further research into the nature of raw drugs named could open up new areas of medicinal plant classification, linking chemistry and bioactivity. Understanding the logic behind the Ayurvedic concept of Abhava Pratinidhi Dravya (drug substitution) could lead to new methods of identifying legitimate drug alternatives, and help solve industry's problems of crude drug shortage.