Emerging role of MicroRNA-Based theranostics in Hepatocellular Carcinoma.

Hepatocellular carcinoma (HCC), with its high mortality and short survival rate, continues to be one of the deadliest malignancies despite relentless efforts and several technological advances. The poor prognosis of HCC and the few available treatments are to blame for the low survival rate, which emphasizes the importance of creating new, effective diagnostic markers and innovative therapy strategies. In-depth research is being done on the potent biomarker miRNAs, a special class of non-coding RNA and has shown encouraging results in the early identification and treatment of HCC in order to find more viable and successful therapeutics for the disease. It is beyond dispute that miRNAs control cell differentiation, proliferation, and survival and, depending on the genes they target, can either promote tumorigenesis or suppress it. Given the vital role miRNAs play in the biological system and their potential to serve as ground-breaking treatments for HCC, more study is required to fully examine their theranostic potential.

RNA Binding Proteins and Non-coding RNA's in Cardiovascular Diseases.

Cardiovascular disease (CVD) is the leading cause of mortality as well as morbidity worldwide. The disease has been reported to be chronic in nature and the symptoms of the disease worsen progressively over a long period of time. Inspite of noteworthy achievements have been made in the therapy of CVD yet the available drugs are associated with various undesirable factors including drug toxicity, complexity, resistance and many more. The versatility of RNAs makes them crucial therapeutics candidate for many human diseases. Deeper understanding of RNA biology, exploring new classes of RNA that possess therapeutic potential will help in its successful translation to the clinic. Understanding the mode of action of various RNAs such as miRNA, RNA binding proteins and siRNA in CVD will help in improved therapeutics among patients. Multiple strategies are being planned to determine the future potential of miRNAs to treat a disease. This review embodies the recent work done in the field of miRNA and its role in cardiovascular disease as diagnostic biomarker as well as therapeutic agents. In addition the review highlights the future of miRNAs as a potential therapeutic target and need of designing micronome that may reveal potential predictive targets of miRNA-mRNA interaction.

Small Interfering RNAs and RNA Therapeutics in Cardiovascular Diseases.

Ribonucleic acid (RNA) is being exploited and understood in its many aspects of function and structure for development of valuable tools in the therapeutics of various diseases such as cardiovascular etc. The expanded knowledge regarding function of RNA in the genomics and inside the cell has dramatically changed the therapeutic strategies in the past few years. RNA has become a spotlight of attention for developing novel therapeutic schemes and hence variety of therapeutic strategies is being coming into the picture that includes RNA interference, use of aptamers, role of microRNA (miRNA) that can alter the complex gene expression patterns. It is due to the fact that RNA offers various advantages in disease management as it can be edited and modified in its various forms such as secondary and tertiary structures. Although scientists are in process of manufacturing RNA-targeting therapies using variety of endogenous gene silencing regulators, Small interfering RNAs (Si RNAs), aptamers and microRNA for cardiovascular diseases yet the development of a novel, risk free therapeutic strategy is a major challenge and need of the hour in cardiovascular medicine. In this regard these agents are required to overcome pleothra of barriers such as stability of drug targets, immunogenicity, adequate binding, targeted delivery etc. to become effective drugs. Recent years have witnessed the progress of RNA therapeutic strategies in cardiovascular diseases that are likely to significantly expand the cardiovascular therapeutic repertoire within the next decade. The present manuscript has been compiled to summarize various approaches of siRNA based therapies in cardiovascular diseases along with the advantages, outcomes and limitations if any in this regard. In addition, the future prospects of RNA therapeutic modalities in cardiovascular diseases are summarized.

Association of CD4/CD8 Ratio with Viral Load, Genotype and Cirrhosis in Chronic Hepatitis C.

ABSTRACT: The interplay between Hepatitis C virus (HCV) and immune system, especially T lymphocytes play a major role in the clearance of virus and in development of liver cell injury resulting in replacement of healthy tissue with fibrous scar tissue. OBJECTIVES: To evaluate the association of CD4/CD8 ratio with viral load and genotype of HCV and to evaluate the correlation of CD4/CD8 ratio and CD4 and CD8 cell counts with liver function tests in HCV infected patients. METHODS: Forty patients of Chronic Hepatitis C infection were enrolled for study. Immunophenotyping by flowcytometry for measurement of CD4 and CD8 T cell counts was used and the percentages of cells expressing CD4 and CD8 were estimated per lymphocyte population. HCV viral load quantitative was done by Roche Taqman Method. RESULTS: The CD4/CD8 ratio was not found to have any significant correlation with HCV viral load. However, it showed a significant difference in the two HCV genotypes, the ratio being higher in genotype 3 than in genotype 1. It showed no significant correlation with liver function tests except serum albumin which had significant positive correlation with CD4/CD8 ratio. The ratio was also found to be significantly decreased in patients with cirrhosis of liver. CONCLUSION: Hepatitis C virus genotype but not viral load influences the immune response to HCV infection. The CD4/CD8 ratio significantly decreases in patients with liver cirrhosis than in normal and fatty liver.

Ethanol content in traditionally fermented ayurvedic formulations: Compromised Good Manufacturing Practice regulations - compromised health.

BACKGROUND: A large part of the population of India prefers the traditional medicine (ayurvedic formulations) for primary health care. However, the effective quality control of herbal medicine is still a big challenge. Numerous reports indicate noncompliance with Compromised Good Manufacturing Practice (GMP) guidelines by the manufacturers which may lead to adverse drug reactions or toxic effects. Asava and arishta are the classical herbal dosage forms wherein fermentation occurs during production leading to the generation of ethanol. The presence of ethanol in these preparations may lead to their misuse. The self-generated ethanol is responsible for extraction of active constituents and acts as a self-preservative. As the procedure for preparation for asava and arishta is same, the ethanol content is also expected to be the same irrespective of the manufacturer. OBJECTIVE: The objective of the present study was to assess and compare the ethanol content of some traditionally fermented ayurvedic formulations available in the market. METHOD: In this study, 20 formulations from 3 different manufacturers available as over-the-counter products were obtained and their ethanol contents were determined using gas chromatograph with flame ionization detector. RESULTS: Statistically significant differences were noted in the ethanol content of various ayurvedic formulations. A simple, less time-consuming, economic, and validated gas chromatographic method for estimation of ethanol in fermented ayurvedic formulations was also developed successfully in present study. CONCLUSION: The data generated during study reflected poor compliance of GMP guidelines by the manufacturers and hence the quality is being grossly compromised posing a safety hazard.

Kitchen phytochemicals from Allium cepa - their role in multidrug resistance.

Bacterial resistance to antibiotics is increasingly becoming a concern to public health. Current antibiotics have failed to bring desirable results to many bacterial infections due to multi-resistant strains. So, concerted efforts are being made to curve this with the help of dietary phytohemicals. A number of dietary phytochemical are being put to trials for antimicrobial activity; however it is worthwhile to search such a part of diet that is very frequently used by major population. Keeping this view in mind an effort has been made to evaluate the antimicrobial activity of most common vegetable component Allium cepa. Methanolic and aqueous extracts were prepared from Allium cepa and antibacterial activity was tested on four pathogenic bacteria (Gram-positive and Gram negative) Staphylococcus aureus, Bacillus Subtilis, Escherichia coli and Pseudomonas aeruginosa using Disc diffusion method and Minimum inhibitory concentration by the Microtitre well plate method. Conventional antibiotic discs of Tetracycline and Gentamycin were used as positive control. Among the two extracts, methanolic extract exhibited a significant antimicrobial activity on the test organisms Ps. aeruginosa (17.5mm) and B. subtilis (11.3mm). The minimum inhibitory concentration value for Ps. aeruginosa was 500µl and for B. subtilis was 250µl for methanolic extract. The growth of other two bacteria i.e. E. coli and S. aureus were not inhibited by methanolic as well as aqueous extract that indicates non susceptibility to aqueous and methanol extract of Allium cepa. From the study, it was concluded that the commonly used Allium cepa possess potent antibacterial property against Bacillus Subtilis and Pseudomonas aeruginosa.

Algal lectins as promising biomolecules for biomedical research.

Lectins are natural bioactive ubiquitous proteins or glycoproteins of non-immune response that bind reversibly to glycans of glycoproteins, glycolipids and polysaccharides possessing at least one non-catalytic domain causing agglutination. Some of them consist of several carbohydrate-binding domains which endow them with the properties of cell agglutination or precipitation of glycoconjugates. Lectins are rampant in nature from plants, animals and microorganisms. Among microorganisms, algae are the potent source of lectins with unique properties specifically from red algae. The demand of peculiar and neoteric biologically active substances has intensified the developments on isolation and biomedical applications of new algal lectins. Comprehensively, algal lectins are used in biomedical research for antiviral, antinociceptive, anti-inflammatory, anti-tumor activities, etc. and in pharmaceutics for the fabrication of cost-effective protein expression systems and nutraceutics. In this review, an attempt has been made to collate the information on various biomedical applications of algal lectins.

Recommendations of apex health bodies remain localized: not in conformity with international implementation policy for urological disorders.

In developing/underdeveloped countries there is still a great burden of adverse drug reaction (ADR), morbidity and mortality because of poor regulations and implementation of preventive measures. These countries try to copy/follow guidelines from international bodies like American Urology Association (AUA), European Association of Urology (EAU), AGS, UMC and WHO irrespective of their country of origin and success in implementation. Although recommendations of these organizations are obligatory, yet these are taken as the gold standard for good clinical practices. This manuscript highlights difference in view point of various apex health organizations in formulating health policies for prevention, diagnosis, treatment and ADR monitoring for urological disorders. Lacking role of regulatory bodies in implementation of existing policies may lead to potentially inappropriate medication and produce a great economic burden. This analysis has prompted us to recommend that these apex bodies should have better coordination in producing a single value document, make it mandatory part of curricula in medical schools for better awareness, awareness campaigns and separate reporting column in ADR form.

Development of a High Penetration Safe Irrigant from Withania somnifera.

Background: Herbal medicine is regaining a strong position in health care by virtue of better safety and minimal toxicity as compared to conventional chemotherapies. It is making a place in dental care in the form of various toothpastes, lotions, and mouthwashes. It has been analyzed that very few discrete herbal irrigants have an action comparable to that of sodium hypochlorite. No data about any study on penetration depth and penetration area for herbal-based irrigants used in root canal treatment are available. Aim: The aim of this study was to assess the efficacy of Withania somnifera extracts by assessing its penetration depth and area inside dentinal tubules using a confocal laser scanning microscope. Materials and Methods: Freshly extracted 25 maxillary central incisors were divided into three groups: Group 1: W. somnifera; Group 2: sodium hypochlorite; and Group 3: distilled water. Results: W. somnifera showed better results with respect to penetration depth and area inside the dentinal tubules thansodium hypochlorite. Conclusion: From the study, it can be concluded that W. somnifera extract-based irrigant may be used as an alternative to sodium hypochlorite-based irrigant. It also shows that more herbal drugs need such types of screening and incorporation in dental practice to reduce the potential side effects of the conventional drugs used in daily practice.

Micronome Revealed miR-205-5p as Key Regulator of VEGFA During Cancer Related Angiogenesis in Hepatocellular Carcinoma.

The high ratio of global mortality rate to incidence rate and steep increase in incidence of liver cancer warrants need for advancement of innovative cancer treatment and therapy for hepatocellular carcinoma (HCC). miRNAs are fascinating prospects as treatments in the form of miRNA mimics or therapeutic targets because of their capacity to target various mRNAs that are changed in diseased states. Micronome is a tool to find signature miRNA for any disease and there is hardly any study on micronome in HCC. The aim of the present study was to identify the genes involved in tumor growth and angiogenesis in HCC patients and determine the signature miRNA by constructing a micronome. Herein, we performed a comprehensive analysis on dysregulated genes obtained from liver cancer gene databases. Only experimentally validated miRNA of angiogenesis genes were included in the study. Micronome was constructed using Cytoscape software and search tools for the retrieval of interacting genes (STRING) database. Dysregulated genes of HCC were integrated with miRNAs for identification of signature miRNA involved and identify genes (acting as positive or negative regulator) to elucidate the potential regulatory pathway or signaling. The study clearly reflects that hsa-mir-205-5p is the signature miRNA for positively regulating angiogenesis in HCC through VEGFA. These regulatory genes and signature miRNAs may be useful to understand the unique angiogenesis process of HCC and quick development of novel/better and cost effective molecular-targeted treatment strategies in HCC as the responsible regulatory molecules can be pinpointed with limited resources with use of bioinformatics.

Bioinformatics Paradigms in Drug Discovery and Drug Development.

Despite breakthroughs in medical sciences, drug development remains a timeconsuming, expensive, challenging, and inefficient process with a high failure rate for novel therapeutic discoveries. Bioinformatics analysis can speed up drug target identification, drug candidate screening, and refining, but it can also help characterise adverse effects and anticipate drug resistance. Integrated genomics, proteomics, and bioinformatics have resulted in potent new tactics for resolving numerous biochemical problems and establishing new methodologies that result in new biomedical products. As a result, a new research trend emerged to demonstrate the mechanism of therapeutic action, forecast drug resistance, and uncover biomarkers for various disorders. The development of new medications is a complicated procedure. There are two basic approaches to drug design: ligand-based drug design and structure-based drug design. The study of protein structure and function was essential for drug development. Current techniques based on combinatorial approaches such as proteomics, genomics, bioinformatics, molecular docking, and mass spectrometry were applied. This article provides an overview of the combinatorial techniques of proteomics, genomics, and bioinformatics that aid in understanding the drug-creation process.

Knowledge, attitude, and practice of medical interns and postgraduate residents on American Geriatric Society updated Beers criteria.

BACKGROUND: Potentially inappropriate medication (PIM) prevalence in older adults is increasing day by day due to lack of knowledge among medical students and professionals. OBJECTIVE: To determine the knowledge of medical students toward American Geriatric Society (AGS) Beers criteria for PIM prescribing. MATERIALS AND METHODS: This cross-sectional study employed a self-administered questionnaire which was validated and designed to assess the knowledge, attitude, and practice of medical interns and postgraduate residents on Beers criteria for PIMs in older adults. The study was conducted in Faridkot region of Punjab in 2019. This study will utilize a purposive sampling strategy and a convenience sampling of up to 183 participants. Mann-Whitney U or Kruskal-Wallis tests were used to compare different issues as appropriate. P values of <0.05 were considered significant. RESULTS: Out of 183 questionnaires distributed, only 155 participants (response rate 84.6%) had filled the survey and were included in the study. 61.3% (n = 95) of the respondents were males. The mean knowledge score of 155 participants was (5.16 ± 1.56), where the highest score was 9 and the lowest score was 2 out of 10. 15.5% (n = 24) of the participant strongly agreed that Beers criteria use is necessary in clinical setting and would be very helpful. While 22.5% (n = 35) of the participants strongly agreed that PIMs cause adverse drug event (ADE) and drug-related problems (DRPs) in older adults. Age of the participant had a significant effect on the knowledge score (p = 0.009), and participant aged between 31 and 40 years had significantly higher knowledge scores compared with participants falls under 20-30 years. CONCLUSION: Medical students and postgraduate residents had average knowledge of PIMs and are unaware of the standard guidelines in older adults such as the Beers criteria. Lack of formal education or training about Beers guidelines was the main reason responsible for average knowledge among participants.

In vitro evaluation of antimicrobial activity of Kutajghan vati - an Ayurvedic formulation.

The present investigation focuses to determine the antimicrobial potential of an Ayurvedic formulation Kutajghan vati. In this study the activity of this formulation was compared with the standard antibiotics like Amikacin and Norfloxacin. Ethanol, methanol and acetone extract of Kutajghan vati demonstrated good antimicrobial activity and thus can form the basis for the development of a novel antibacterial formulation.

Chronic Kidney Disease (CKD) - A Brand Ambassador/Alarming Bell for Potentially Inappropriate Medication in Elderly Inpatients.

BACKGROUND: Since the past decade, prevalence of Potentially Inappropriate Medication (PIM) among elderly inpatients has increased drastically. However, limited data is available on PIM indicators and PIMs use among the elderly in patients suffering from Chronic Kidney Disease (CKD). OBJECTIVE: The objective of this study is to determine the prevalence of PIMs in elderly hospitalized patients with CKD. METHODS: A cross-sectional study was carried out on 102 patients in a tertiary care hospital. PIMs were identified using Beers criteria 2019. Chi-square test was used to identify the association between variables and PIMs use. RESULTS: PIMs, as assessed according to AGS updated Beers criteria 2019 was found to be in more than 68.6% of patients of median age 65 years and 3 diagnoses and 7 days median length of stay. Most of the patients (47.1%) had ≥4 diagnosis. The most common comorbidities in patient were diabetes mellitus (n=54) and hypertension (n=55). Most of the subjects (66.7%) were on polypharmacy (5-9 medications/day) and 25.5% were on a higher level of polypharmacy (>10 medicines/day). Approximately 90% of the patients were having very low CrCl < 21ml/min (calculated with the help of Cockcroft- Gault formula). A significant association between PIM use and an increased number of diagnoses, polypharmacy or high-level polypharmacy, was observed. CONCLUSION: The prevalence of PIMs in elderly inpatients suffering from CKD is quite high. The study clearly indicates negligence/ lack of awareness amongst physicians leading to increase in PIM use. Authors propose that the CKD patients should attract special attention of physician and should be treated as brand ambassadors/alarming bell for PIM use.

Erdafitinib as a Novel and Advanced Treatment Strategy of Metastatic Urothelial Carcinoma.

Urothelial carcinoma has become the ninth most common malignancy in the world. Since the 1980s, diverse studies and treatment methods came out with their possible effects along with certain limitations. Initially, platinum chemotherapy was considered as first-line treatment of the disease. Although it was proved to be effective initially, the most number of cases reported the reoccurrence of the disease. Furthermore, aberrant ligand- dependent and constitutive ligand-independent fibroblast growth factor receptor (FGFR) signaling has been reported in a large number of solid tumors, including urothelial carcinoma that became the basis for FGFR inhibition for the treatment of the disease. Erdafitinib is a pan-FGFR inhibitor that was recently approved in the USA for the treatment of locally advanced or metastatic FGFR3 or FGFR2 urothelial carcinoma. The drug is also being investigated as a treatment for other cancers, including cholangiocarcinoma, liver cancer, non-small cell lung cancer, prostate cancer, lymphoma cancer and oesophageal cancer. This article summarizes the various treatments that evolved for bladder cancer till now, a brief description of the biology of FGFR inhibition, clinical pharmacology, and various clinical trials of erdafitinib.

Prevalence and predictors of Potentially Inappropriate Psychotropic Medication in older adults with psychiatric illness.

BACKGROUND: There is limited information available on the use of Potentially Inappropriate Psychotropic (PIP) medication in older adults having a psychiatric illness. OBJECTIVE: To determine the prevalence of and predictors of prescribing PIP medications in older adults with psychiatric illness. METHODS: A cross-sectional study was carried out at a tertiary care hospital on 456 patients of either sex, with a median age of 65 years attending the psychiatry outpatient department. Evaluation of PIP medication was done using Beers criteria 2019 and Screening tool of Older Persons Potentially Inappropriate Medication (STOPP) criteria 2015. Bivariate logistic regression was used to find out the predictors of PIP prescribing. RESULTS: Results of the study reflect that a staggering number of older adults, (more than 91% and 73%) out of a total of 456 patients were prescribed with at least one PIP medication identified by Beers criteria and STOPP criteria, respectively. Long-acting benzodiazepine (LABZD) like clonazepam was identified as one of the most commonly prescribed PIP medications by both sets of criteria. Further analysis revealed that older adults from rural background (Odds Ratio (OR) 2.60, 95% Confidence Interval (CI) 1.20-5.65; P = 0.015), Tricyclic Antidepressant (TCA) (OR 0.30, 95% CI 0.12-0.75; P = 0.010), LABZD (OR 33.72, 95% CI 11.27-100.85; P = < 0.001), atypical antipsychotics (OR 22.35, 95% CI 5.31-93.99; P = < 0.001) use were most common predictors for PIP medication prescribing. CONCLUSION: The study suggests that the Beer criteria detects more PIP medication than the STOPP criteria.

Clonazepam tops the list of potentially inappropriate psychotropic (PIP) medications in older adults with psychiatric illness: A cross-sectional study based on Beers criteria 2019 vs STOPP criteria 2015.

BACKGROUND: In older adults, polypharmacy and potentially inappropriate psychotropic (PIP) medication use are prominent prescription challenges. However, there is limited information available on the use of PIP medication in older adults having psychiatry illness. OBJECTIVE: To find out the most commonly prescribed PIP in tertiary care hospitals of developing countries with respect to Beers criteria 2019 and Screening Tool of Older Persons' Potentially Inappropriate Prescriptions (STOPP) and predictors of PIP. METHODS: A cross-sectional analysis of 456 patients of either sex with a median age of 65 years visiting the outpatient department of psychiatry was performed at the tertiary care hospital of North India with respect to Beers criteria 2019 and STOPP criteria 2015. Bivariate logistic regression was used to figure out the predictors of PIP medication. RESULTS: Results of the study reflects a staggering number of older adults, (more than 91 % and 73 %) out of total 456 patients were prescribed with at least one PIP medication as per Beers criteria and STOPP criteria, respectively. Long-acting benzodiazepine like clonazepam, chlordiazepoxide were identified as one of the most commonly prescribed PIP medications with respect to the both set of criteria. Further analysis revealed that polypharmacy (≥5 medications with odds Ratio (OR) 17.33, 95% Confidence Interval (CI) 1.42-210.66, P-0.025) as the sole important predictor for PIP medication. CONCLUSION: According to the Beers criterion and the STOPP criteria, the use of PIP medicine is very prevalent among older adults with psychiatric illness. The Beers criteria dramatically diagnose more PIP medication than STOPP criteria.

Integration of System Biology Tools to Investigate Huperzine A as an Anti-Alzheimer Agent.

Aim: The present study aimed to investigate huperzine A as an anti-Alzheimer agent based on the principle that a single compound can regulate multiple proteins and associated pathways, using system biology tools. Methodology: The simplified molecular-input line-entry system of huperzine A was retrieved from the PubChem database, and its targets were predicted using SwissTargetPrediction. These targets were matched with the proteins deposited in DisGeNET for Alzheimer disease and enriched in STRING to identify the probably regulated pathways, cellular components, biological processes, and molecular function. Furthermore, huperzine A was docked against acetylcholinesterase using AutoDock Vina, and simulations were performed with the Gromacs package to take into account the dynamics of the system and its effect on the stability and function of the ligands. Results: A total of 100 targets were predicted to be targeted by huperzine A, of which 42 were regulated at a minimum probability of 0.05. Similarly, 101 Kyoto Encyclopedia of Genes and Genomes pathways were triggered, in which neuroactive ligand-receptor interactions scored the least false discovery rate. Also, huperzine A was predicted to modulate 54 cellular components, 120 molecular functions, and 873 biological processes. Furthermore, huperzine A possessed a binding affinity of -8.7 kcal/mol with AChE and interacted within the active site of AChE via H-bonds and hydrophobic interactions.

Estimation of heavy metals in commonly used medicinal plants: a market basket survey.

Popularity of herbal drugs is increasing all over the world because of lesser side effects as compared to synthetic drugs besides it cost effectiveness and easy availability to poor people particularly in developing countries. Keeping in view the increased market demand of herbal drugs, it is essential to ensure their chemical quality prior to use. Raw drugs and herbs are usually collected from different places, which might be contaminated with various contaminants. It is pertinent to estimate the levels of heavy metals and other micronutrients, which could be affected by their presence in the surrounding environments. Heavy metals are known to pose a potential threat to terrestrial and aquatic biota. Keeping this in view, samples of ten plants or plant parts used in drug making were collected from local markets of Punjab for heavy metal and micronutrient estimation. It was found that the samples were contaminated having cadmium, lead, chromium, iron, manganese, and zinc. The highest mean level of cadmium (23.1 µg/kg) was found in Haritaki sample. Chromium concentration of the plant samples ranged between 7.25 and 1.34 µg/kg with the highest values was in Daruharidra and lowest in Pippali. The levels of these heavy metals were within permissible limits.

Ayurvedic formulations containing benzoic and ascorbic acids as additives: benzene formation during storage and impact of additives on quality parameters.

OBJECTIVES: Ayurvedic formulations are becoming the prior choice of people as health care supplements. The increasing demand for these formulations has led to extensive development of Ayurvedic pharmaceutical industries worldwide. The reaction between the preservatives (sodium benzoates and ascorbic acid) used in these formulations could generate benzene. Benzene is classified as class-1 human carcinogen and responsible for various short and long term health effects. METHODS: In this study, 25 formulations (containing ascorbic acid and sodium benzoate) of various manufacturers available as over the counter products were obtained and their benzene content were determined using gas chromatograph with flame ionization detector. RESULTS: The result showed that 64% of the formulations were free from benzene contamination whereas 36% of formulations were found to be contaminated with benzene. A simple, less time-consuming, economic, and validated gas chromatographic method for estimation of benzene in Ayurvedic formulations was also developed successfully in present study. CONCLUSIONS: The data revealed that the level of benzene was within permissible limits, yet the presence of a carcinogen in the marketed formulations intended for internal use is an alarming situation.