Different mechanisms of arsenic related signaling in cellular proliferation, apoptosis and neo-plastic transformation.

Arsenic is a toxic heavy metal vastly dispersed all over the earth crust. It manifests several major adverse health issues to millions of arsenic exposed populations. Arsenic is associated with different types of cancer, cardiovascular disorders, diabetes, hypertension and many other diseases. On the contrary, arsenic (arsenic trioxide, As2O3) is used as a chemotherapeutic agent in the treatment of acute promyelocytic leukemia. Balance between arsenic induced cellular proliferations and apoptosis finally decide the outcome of its transformation rate. Arsenic propagates signals via cellular and nuclear pathways depending upon the chemical nature, and metabolic-fates of the arsenical compounds. Arsenic toxicity is propagated via ROS induced stress to DNA-repair mechanism and mitochondrial stability in the cell. ROS induced alteration in p53 regulation and some mitogen/ oncogenic functions determine the transformation outcome influencing cyclin-cdk complexes. Growth factor regulator proteins such as c-Jun, c-fos and c-myc are influenced by chronic arsenic exposure. In this review we have delineated arsenic induced ROS regulations of epidermal growth factor receptor (EGFR), NF-ĸß, MAP kinase, matrix-metalloproteinases (MMPs). The role of these signaling molecules has been discussed in relation to cellular apoptosis, cellular proliferation and neoplastic transformation. The arsenic stimulated pathways which help in proliferation and neoplastic transformation ultimately resulted in cancer manifestation whereas apoptotic pathways inhibited carcinogenesis. Therapeutic strategies against arsenic should be designed taking into account all these factors.

Arsenic Induced Oxidative Neural-Damages in Rat are Mitigated by Tea-Leave Extract via MMPs and AChE Inactivation, Shown by Molecular Docking and in Vitro Studies with Pure Theaflavin and AChE.

BACKGROUND: Chronic arsenic-exposure causes neuromuscular disorders and other health anomalies. Damage to DNA and cytoskeletal/extracellular matrix is brought on by reactive-oxygen-species (ROS)-induced intrinsic antioxidant depletion (thiols/urate). Therapeutic chelating-agents have multiple side-effects. OBJECTIVES: The protection of (Camellia sinensis) tea-extract and role of uric-acid (UA) or allopurinol (urate-depletor) on arsenic-toxicity were verified in rat model. METHODS: Camellia sinensis (CS dry-leaves), UA or allopurinol was supplemented to arsenic-intoxicated rats for 4-weeks. Purified theaflavins and their galloyl-ester were tested in-vitro on pure AChE (acetylcholinesterase) and their PDB/PubChem 3-D structures were utilized for in-silico binding studies. The primary chemical components were evaluated from CS-extracts. Biochemical analysis, PAGE-zymogram, DNA-stability comet analysis, HE-staining was performed in arsenic-exposed rat brain tissues. RESULTS: Animals exposed to arsenic showed symptoms of erratic locomotion, decreased intrinsic antioxidants (catalase/SOD1/uric acid), increased AChE, and malondialdehyde. Cerebellar and cerebrum tissue damages were shown with increased levels of matrix-metalloprotease (MMP2/9) and DNA damage (comets). Allopurinol- supplemented group demonstrated somewhat similar biochemical responses. In the CS-group brain tissues especially cerebellum is considerably protected which is evident from endogenous antioxidant and DNA and cytoskeleton protection with concomitant inactivation of MMPs and AChE. Present study indicates theaflavin-digallate (TFDG) demonstrated the highest inhibition of purified AChE (IC50 = 2.19 µg/ml with the lowest binding free-energy; -369.87 kcal/mol) followed by TFMG (IC50 = 3.86 µg/ml, -347.06 kcal/mol) suggesting their possible restoring effects of cholinergic response. CONCLUSIONS: Favorable responses in UA-group and adverse outcome in allo-group justify the neuro-protective effects of UA as an endogenous antioxidant. Role of flavon-gallate in neuro protection mechanism may be further studied.

Avena sativa (Oat), a potential neutraceutical and therapeutic agent: an overview.

The aim of the present review article is to summarize the available information related to the availability, production, chemical composition, pharmacological activity, and traditional uses of Avena sativa to highlight its potential to contribute to human health. Oats are now cultivated worldwide and form an important dietary staple for the people in number of countries. Several varieties of oats are available. It is a rich source of protein, contains a number of important minerals, lipids, ß-glucan, a mixed-linkage polysaccharide, which forms an important part of oat dietary fiber, and also contains various other phytoconstituents like avenanthramides, an indole alkaloid-gramine, flavonoids, flavonolignans, triterpenoid saponins, sterols, and tocols. Traditionally oats have been in use since long and are considered as stimulant, antispasmodic, antitumor, diuretic, and neurotonic. Oat possesses different pharmacological activities like antioxidant, anti-inflammatory, wound healing, immunomodulatory, antidiabetic, anticholesterolaemic, etc. A wide spectrum of biological activities indicates that oat is a potential therapeutic agent.

Evaluation of the anti-arthritic activity of Rhuflex-F - A proprietary Ayurvedic herbomineral formulation in albino rats.

Background: Rhuflex-F is a proprietary Ayurvedic herbo-mineral formulation clinically used to combat and relieve stiffness in joints and muscles, reduce edema, restore mobility, and also effective in relieving the symptoms of other autoimmune illnesses that lead to rheumatism. Aims: The aim and objective of the research study is to evaluate the efficacy of Rhuflex-F against in vitro protein denaturation and in vivo Freund's adjuvant-induced arthritis in albino rats. Materials and methods: In vitro inhibition of protein denaturation activity was carried out using bovine serum albumin. For in vivo activity, arthritis was induced by complete Freund's adjuvant in albino rats. Rhuflex-F (135-270 mg/kg, po) was administered for 30th days in arthritic rats, and effects were assessed on primary and secondary paw edema, on pain response, hematological, serum biochemical parameters (serum transaminases, alkaline phosphatase, urea, uric acid, and orosomucoid), and serum anti-oxidant parameters and adrenal ascorbic acid. Results: Aqueous extract of Rhuflex-F showed in vitro protein denaturation inhibitory activity in a dose-dependent manner. Rhuflex-F showed nonsignificant decrease in primary and secondary paw edema with reduced pain response, some reversal effects on hematological parameters such as white blood cell and red blood cell related parameters and serum orosomucoid and adrenal ascorbic acid in comparison to Fruend's adjuvant control group. Further, Rhuflex-F reversed Freund's adjuvant-induced adverse effects on oxidant status in the serum of albino rats. Conclusion: Result of the present study suggested that Rhuflex-F formulation has anti-inflammatory activity, may be due to the inhibition of protein denaturation in vitro and in vivo anti-arthritic activity against complete Freund's adjuvant-induced arthritis in albino rats.

Relation of ACE2 with co-morbidity factors in SARS-CoV-2 pathogenicity.

In the last 3 years of the pandemic situation, SARS-CoV-2 caused a significant number of deaths. Infection rates for symptomatic and asymptomatic patients are higher than that for death. Eventually, researchers explored that the major deaths are attributed to several comorbidity factors. The confounding factors and gender-associated infection/death rate are observed globally. This suggests that SARS-CoV-2 selects the human system recognizing the internal comorbid environment. This article explored the influences of hypertension, diabetes, cardiovascular, and renovascular disorders in COVID-19 severity and mortality. Brief mechanistic layouts have been presented here, indicating some of the comorbidity as the critical determinant in the COVID-19 pathogenesis and related mortality.

ACE/ACE2 balance might be instrumental to explain the certain comorbidities leading to severe COVID-19 cases.

The outbreak of Severe Acute Respiratory Syndrome Coronavirus-2 (SARS-CoV-2) is a global catastrophe. The elderly and people with comorbidity are facing a serious complication of the disease. The entry and infection strategy of SARS-CoV-2 in a host cell is raised by an amazing way of angiotensin-converting enzyme (ACE) 2 (ACE2) receptor recognition and imbalance of ACE/ACE2 in various organs, especially in the lungs. Here it has been discussed the role of interferon and protease during the receptor recognition (begining of infection) and followed by the impact of cytokine and hypoxia in the context of the balance of ACE/ACE2. It has also very concisely delineated the biochemistry and mechanism of ACE/ACE2 balance in different stages of infection and its role in comorbidity.

Functional Role of the Rodlet Cell and Macrophage in Neural Protection of the Olfactory Neuroepithelium in a Teleostean: Gobiid (Pseudapocryptes lanceolatus [Bloch and Schneider, 1801]): An Ultrastructural Study.

BACKGROUND: Neural protection of the olfactory epithelium against pathogenic invasion is still hardly addressed in fish chemosensory research. AIMS AND OBJECTIVES: The ultrastructural detail on the rodlet cell and macrophage has been studied within the olfactory neuroepithelium of Pseudapocryptes lanceolatus to correlate their role in the neural protection of the chemosensory system. MATERIALS AND METHODS: The cellular structures were examined under light microscope (LM: Primo Star; Carl Zeiss Microscopy, GmbH, Germany) and transmission electron microscope (Morgagni 268D). RESULTS: Three distinct stages of the rodlet cell (viz., immature, mature, and degenerative) and macrophages have been characterized at the various depths of the olfactory neuroepithelium in P. lanceolatus. The cytoarchitecture of degenerative rodlet cell indicates holocrine mode of secretion against pathogenic invasion into the nasal cavity. Macrophages possess prominent pseudopodia, extending toward invading pathogens. The interaction between macrophage and invading pathogens implicates the role of macrophage as a scavenger to eliminate the pathogens by phagocytosis from the neuroepithelial system. CONCLUSION: This study denotes a significant difference in the mode of action of rodlet cell and macrophages, but they are commonly involved in cell-mediated nonspecific immune response against the invading pathogens.

Safety profile of Ayurvedic poly­herbomineral formulation - Bacnil capsule in albino rats.

INTRODUCTION: Bacnil capsule is a poly­herbomineral formulation used to treat gastroenteritis. It contains many potential drugs derived from plant sources and Bhasma (calcined fine powder) preparations. AIMS: The study was designed to ascertain the safety of bacnil capsule orally in Charle's Foster albino rats. MATERIALS AND METHODS: As per the Organization for Economic Cooperation and Development (OECD) 425 protocol oral acute toxicity study, bacnil capsule was administered orally once only at the dose of 2000 mg/kg in rats. For repeated dose toxicity study, AYUSH 170 and OECD 407, it was administered at three dose levels, Therapeutic doses (TED) (196.2), TED × 5 (981) and TED × 10 (1962) mg/kg/day orally for 28 days in albino rats followed by a 15­day recovery period only on TED × 10 dose level. OBSERVATION AND RESULTS: Bacnil at the oral dose of 2000 mg/kg did not produce any toxicity or mortality in albino rats. Repeated dose 28­day oral toxicity revealed that test formulation did not produce any significant change in serum biochemical, hematological, and histopathological parameters at therapeutic dose level. Mild­to­moderate pathological changes were observed in the various serum biochemical and cytoarchitecture of the liver, heart, kidney, and stomach at a dose of 10 TEDs; however, the same was reversed after discontinuation in the recovery test. CONCLUSION: Bacnil at 196.2 mg/kg/day is safe at the therapeutic dose level in albino rats.

Insulin resistance in prostate cancer patients and predisposing them to acute ischemic heart disease.

Lack of insulin or insulin resistance (IR) plays a central role in diabetes mellitus and makes diabetics prone to acute ischemic heart disease (AIHD). It has likewise been found that many cancer patients, including prostate cancer patients die of AIHD. Previously it has been delineated from our laboratory that dermcidin could induce anomalous platelet aggregation in AIHD and also impaired nitric oxide and insulin activity and furthermore dermcidin was also found in a few types of cancer patients. To determine the role of this protein in prostatic malignancy, a retrospective case-control study was conducted and blood was collected from prostate cancer patients and healthy normal volunteers. So, we measured the level of dermcidin protein and analyzed the IR by Homeostasis Model Assessment (HOMA) score calculation. Nitric oxide was measured by methemoglobin method. HDL, glycated hemoglobin (HbA1c), BMI, hs-cTroponin-T were measured for the validation of the patients' status in the presence of Dermcidin isoform-2 (DCN-2). Multiple logistic regression model adjusted for age and BMI identified that the HOMA score was significantly elevated in prostate cancer patients (OR = 7.19, P<0.001). Prostate cancer patients are associated with lower level of NO and higher level of both proteins dermcidin (OR = 1.12, P<0.001) and hs-TroponinT (OR = 1.76, P<0.001). From the results, it can be interpreted that IR plays a key role in the pathophysiology of prostate cancer where dermcidin was the cause of IR through NO inhibition leading to AIHD was also explained by high-sensitive fifth generation cTroponin-T (hs-cTroponinT) and HbA1c level which are associated with endothelial dysfunction.

Ultrastructure Based Morphofunctional Variation of Olfactory Crypt Neuron in a Monomorphic Protogynous Hermaphrodite Mudskipper (Gobiidae: Oxudercinae) (Pseudapocryptes lanceolatus [Bloch and Schneider]).

Pseudapocryptes lanceolatus (Bloch and Schneider) is a monomorphic protogynous hermaphrodite teleost that possesses ovotestis as gonadal unit of reproductive structure. At the onset of breeding season (i.e., June-July), the ovarian tissue is gradually differentiating into female-phased P. lanceolatus. At the same time, the pear-shaped crypt cells (a type of neuron) are frequently appeared at apical part of pseudostratified olfactory neuroepithelium of P. lanceolatus. The crypt neuron is characterized by the presence of sunken cilia and microvilli at the proximal region. The features of subcellular organelles are also explored in lieu of their probable functional significance. The nucleoplasm of mature crypt neuron shows chromatin granules having diameter: 15-25 nm. This cell undergoes neural apoptosis at the end of breeding phase (i.e., October-November). Fragmented chromatin fibers with numerous chromatin granules (diameter: 25-30 nm) in nucleoplasm and lysosomal diversity are the most notable characters of apoptotic crypt neuron. The large accumulation of heterochromatin chromatins in nucleoplasm is also marked under fluorescence microscope. The frequent presence of acetylcholinesterase-positive vesicles in axoplasm of crypt neurons is also a prime subcellular indicator for inhibition of neural transmission of olfactory signals. Therefore, it is concluded that the sex differentiation in P. lanceolatus and occurrence of crypt neuron in olfactory neuroepithelium are interrelated events during the reproductive period. Consequently, we hypothesized that the crypt neuron plays an active role in the implementation of unique reproductive strategy through recognition of pheromonal cues within the social organization of P. lanceolatus.

Acute toxicity and repeated dose 28-day oral toxicity study of metriviv syrup in female rats.

INTRODUCTION: Metriviv syrup is a poly-herbal formulation used as uterine tonic and for treating gynecological ailments such as infertility, leucorrhea, and menstrual disorders. There is no scientific data on the safety of this formulation available, therefore, its detail toxicity study in female albino rats was conducted. AIMS AND OBJECTIVE: Acute toxicity and repeated dose 28 days oral toxicity study of metriviv syrup in female rats. MATERIAL AND METHODS: In oral acute toxicity study, syrup metriviv was administered orally once only at the dose of 2000mg/kg in rats in a sequential manner. For repeated dose toxicity study, Metriviv syrup was administered orally for 28 consecutive days in albino rats at three dose levels, i.e., TED, TEDx5 and TEDx10 dose levels (1.35, 0.75, 13.5 ml/kg, twice a day respectively). RESULTS AND OBSERVATION: Results of acute toxicity study showed that drug did not produce any behavioral changes, signs of toxicity and mortality up to the dose of 2000 mg/kg in rats. In repeated dose 28-day oral toxicity study, Metriviv did not produce any signs of toxicity and changes in behavioral parameters. Metriviv did not affect cellular as well as non-cellular elements of the blood to significant extent. Serum biochemical profile and histological study clearly indicated that the test formulation is not likely to produce any serious changes at lower dose level while produced mild-to-moderate changes at TEDx10 dose level and same was reverted in recovery study after discontinuation of the drug. CONCLUSION: The study concluded that metriviv syrup is safe to administer up to dose of 2000 mg/kg in female rats during acute toxicity. In repeated dose 28-day oral toxicity study, test drug at TEDx10 has only mild-to-moderate adverse potential for liver and kidney while did not have any major toxic effects at therapeutic dose level in female albino rats.

Attenuation of utero-toxicity, metabolic dysfunction and inflammation by soy protein concentrate in rats exposed to fluoridated water: consequence of hyperlipidemia in parallel with hypohomocysteinemia.

Lipid peroxidation and ROS generation are the pathogenesis of chronic fluoride toxicity, and its detrimental effects on human reproduction are noted drastically. The aim of the present study was to elucidate the defensive effects of soy protein concentrate (SPC) against sodium fluoride (NaF)-induced uterine dysfunction at biochemical and histological level. Rats were randomly distributed into four groups as control, NaF-treated (200 ppm), and SPC co-administered groups (20 mg and 40 mg/ 100 g body weight) for 16 days. SPC reversed the toxic effects of NaF. SPC significantly ameliorated the NaF-induced alterations of the antioxidant system in the uterus by decreasing lipid peroxidation products and by increasing antioxidant activities. SPC significantly counteracted the adverse effects of NaF on serum level of lactate dehydrogenase (LDH) and inflammatory markers Interleukin-6 (IL-6), tumor necrosis factor alpha (TNF-α) and nuclear factor kappa-B (NF-κB). Our results also explored that lipid profile was meaningfully altered due to NaF and also focused a diminution of circulating homocysteine (Hcy) and altered lipid profiles along with a diminished quantity of serum B12 and B9. However, both the doses of SPC reverted back serum levels of B12, B9, and Hcy status in similar fashion along with its corrective action on lipid profile. NaF-treated group exhibited a marked degree of reduction in the weights of ovary and uterus with an alteration of normal tissue histology and significant diminution in serum estradiol (ES) levels without fluctuating uterine estradiol receptor-α (ER-α). However, SPC restored the normal tissue histoarchitecture and also increased the functional efficiency and expression of the ER-α receptor by overturning the ES levels in NaF-treated rats. Moreover, both the doses of SPC were effective against NaF-induced alterations, although 40 mg SPC/100 g body weight had better efficacy in ameliorating the NaF-induced adverse effects on the uterus and ovary.

Estriol Inhibits Dermcidin Isoform-2 Induced Inflammatory Cytokine Expression Via Nitric Oxide Synthesis in Human Neutrophil.

BACKGROUND: An increase in the level of cytokines like TNF-α and IL-6 causes the inflammatory surge in acute ischemic heart disease (AIHD). OBJECTIVE: A high-level dermcidin isoform-2 (DCN-2) occurrence in AIHD was subjected to determine a possible regulation of cytokines expression. The effect of estrogen to counteract the inflammatory response was determined. METHODS: Blood was collected from AIHD patients and normal volunteers with consent. Nitric oxide (NO) synthesis was done with methemoglobin method.TNF-α and IL-6 expression were determined by ELISA and Western blot. RESULTS: (DCN-2) incubation with 120nM to the normal neutrophil solution for 2h resulted in the increase of TNF-α from 3.82±1.53pg/ml to 20.7±6.9pg/ml and IL-6 from 3.27±1.52pg/ml to 47.07±3.4pg/ml. In AIHD patients, the cytokine level was18.3- 27.3pg/ml, with a median value 21.86pg/ml (TNF-α) and IL-6 level was 23.54- 52.73pg/ml, with a median value 42.16pg/ml. Treatment with 0.6nM estriol, a kind of female steroid hormone estrogen for 45min decreased the elevated cytokine level in 120nM DCN-2 treated normal neutrophils. DCN-2 induced TNF-α synthesis in neutrophils was further determined by Western blot technique with a thickened band intensity of TNF-α. Estriol (0.6nM) treatment also influenced the DCN-2 induced inhibition of nitric oxide (NO) synthesis from 0nmol NO/ml to 0.56nmol/ml. The subsequent reduction of TNF-α level correlates the increase of NO level. CONCLUSION: In conclusion, the stress-induced DCN-2 production in AIHD propagates the inflammatory response. Steroid molecule like estriol plays a protective role by reducing DCN-2 responses in the NO synthesis.

Electron microscope based X-ray microanalysis on bioaccumulation of heavy metals and neural degeneration in mudskipper [Pseudapocryptes lanceolatus].

The bioaccumulation of heavy metals and its probable cytological consequences in ciliated olfactory sensory receptor neuron (OSN) of two different groups of Pseudapocryptes lanceolatus has been studied using X-ray microanalysis in transmission electron microscopy (TEM-EDX) [i.e., Group I, collected near Kanchrapara (22.56°N 88.26°E) and Group II, collected near Tribeni (22.99°N 88.40°E) of West Bengal, India]. The ciliated OSN is a bipolar neuron and possesses a prolonged dendron with four to six cilia at the olfactory knob, perikaryon, and axon. Excess accumulation of copper (94.50%) and iron (83.81%) was noted under TEM-EDX in the cytoplasm of the olfactory knob as well as nucleoplasm of ciliated OSNs in P. lanceolatus (Group II). The degenerating ciliated OSNs show distinct features of lysis of the plasma membrane at the olfactory knob, disintegration of cytoskeletal structures in perinuclear cytoplasm and axoplasm, and fragmented chromatin fibers with granules (diameter, 20-30 nm) in the nucleoplasm. Crowding of acetylcholinesterase-positive vesicles (diameter:, 30-40 nm) at the terminal part of the axoplasm was related to accumulation of heavy metals in degenerating ciliated OSNs of P. lanceolatus (Group II). The recorded concentrations of heavy metals in the same organ among different groups of P. lanceolatus in varying geographical areas indicates the stress of concerned environmental health. This ultrastructural interpretation on the fish ciliated OSN is a prerequisite for monitoring environmental health as well as metallobiology of several neurodegenerative disorders in fish caused by bioaccumulation of heavy metals.

Detection of tallow adulteration in cow ghee by derivative spectrophotometry.

CONTEXT: Ghee is a widely consumed dairy product in India and that prepared from cow milk is mentioned in ayurvedic texts as an ingredient of many formulations/additive as well. Detection of cow ghee adulteration with vegetable oils/fats and animal body fats is a key concern. Indicated values for commonly used parameters to differentiate pure and adulterated ghee materials are many a times overlapping. Among reported techniques, ultraviolet fluorescence and paper chromatography technique are not that much sensitive while other methods require sophisticated instrumental facilities (such as gas chromatography, mass spectrometry) and costly analytical processes. AIMS: The present paper deals with a promising spectroscopic method to determine the tallow adulteration in cow ghee. MATERIALS AND METHODS: Ghee and tallow (taken in chloroform) as such and mixed in different proportions were scanned by spectrophotometer and their second order spectra were analyzed. RESULTS: The value of the ratio of the absorbance of peaks at about 238 nm and 297 nm steadily decreases with the increasing proportion of tallow. This decrease shows consistent linearity suggesting its applicability for quantitative estimation of tallow in cow ghee. CONCLUSION: The developed derivative spectroscopic method is a rapid, sensitive, cost-effective method for detection of tallow adulteration in cow ghee.

Trividha Snehapaka of Panchagavya Ghrita: A critical comparative evaluation.

BACKGORUND: The three stages of Snehapaka formulations namely Mridu, Madhyama and Khara Paka have been characteristically advocated for different routes of administration-Nasya, Pana/Basti and Abhyanga, respectively. Guidelines or established method for post-formulation characterization for the same is hardly available. OBJECTIVE: The present communication is the comparative study of Mridu, Madhyama and Khara Paka of Panchagavya ghrita (PGG). MATERIALS AND METHODS: Laboratory prepared samples of PGG following classical method were analyzed for different physicochemical, spectroscopic, chromatographic parameters, and antioxidant activity. RESULTS: No significant difference was found among Mridu, Madhyama and Khara Paka in physicochemical parameters as well as chromatographic profiles. The ratio of absorbance at 240 and 294 nm showed steady increase from Mridu to Madhyama to Khara Paka in the ultraviolet (UV)-visible spectra of unsaponifiable matter. The high performance thin layer chromatography (HPTLC)-2,2 Diphenyl-1-picryl hydrazil (DPPH) bioautography assay revealed presence of two antioxidant compounds in low concentration in all the samples. This was further supported by estimation of total reducing power and DPPH assay. No significant difference was found among the three samples. CONCLUSION: Comparison of various physicochemical parameters, chromatographic profiles, and in vitro antioxidant activity determination is of little help in establishing any significant difference among the samples. However, spectrophotometric analysis of unsaponifiable matter reveals some encouraging characteristic findings which will be useful in establishing difference among the three stages of processing of PGG as well as Snehapaka in general.

Clinical efficacy of "Bhadra Mustadi Paste" and "Nagaradi Kwatha Gandusha" in Shitada (Gingivitis).

Shitada is one of the Dantamulagata Rogas mentioned by Acarya Susruta under the heading of Mukha Rogas. It is characterized by symptoms like spontaneous bleeding, halitosis, blackish discoloration of gums, and so on. The symptomatology of Shitada can be compared with gingivitis, which is caused mainly by the accumulation of debris, plaque, and calculus at the tooth margin due to ignorance of oral care. It can progress into periodontitis - the disease in which the firmness, contour, and position of the gums are altered and teeth become mobile. The prevalence of gingivitis is as high as 50% in most of the population. In the present study, a total of 33 patients were registered in three groups. Bhadramustadi yoga has been used in paste form for local application along with Nagaradi Kwatha Gandusa in Group A and in powder form for Pratisarana along with Nagaradi Kwatha Gandusa in Group B. In Group C (Control) Sphatika Jala was used. The patients were diagnosed and assessed on the basis of Ayurvedic as well as modern classical signs and symptoms. Further assessment was done with the different indices used in modern dentistry. All the signs and symptoms were given a score depending upon the severity, to assess the effect of the drugs objectively. The effect of therapy was highly significant in group A in most of the signs, symptoms, and indices; and in group B it was highly significant in some signs and symptoms.