Anti-skin ageing activity of napthoquinones from Arnebia nobilis Reichb.f.

The present isolation and identification of napthoquinones from roots of Arnebia nobilis Reichb.f. can lead to the discovery of new anti-skin ageing ingredient in colour cosmetics. Four compounds have been isolated and purified by rigorous column chromatography. The compounds are identified as ß, ß-dimethylacryl alkannin (AN-I), acetoxyisovaleryl alkannin (AAN-II), acetyl alkannin (AN-III) and alkannin (AN-IV) by interpretation of spectroscopic data. This study is the first to report the isolation of Acetoxyisovaleryl alkannin (AAN-II) from A. nobilis. The IC50 values of the compounds, determined in human skin cells (human dermal fibroblasts and human keratinocytes) and mouse embryonic fibroblasts (NIH3T3) varied significantly among the four alkannins. Among the four compounds, ß-acetoxyisovaleryl alkannin (AAN-II) significantly inhibited hydrogen peroxide (H2O2)-induced red blood corpuscle haemolysis and cellular senescence in human dermal fibroblasts. Collagen-I, elastin and involucrin syntheses in human dermal fibroblasts or keratinocytes were up regulated by AAN-II. These results support the potential utility of alkannins as novel anti-ageing ingredients.

Yashada bhasma (Zinc calx) and Tankana (Borax) inhibit Propionibacterium acne and suppresses acne induced inflammation in vitro.

OBJECTIVE: Yashada bhasma (YB) and Tankana (TA) are well characterized minerals used in traditional medicine for the treatment of various skin ailments. Yashada bhasma and TA are a unique preparation of zinc and borax, respectively. The study was conducted to evaluate the in vitro inhibitory effect of YB, TA and its combination (YBTA) on Propionibacterium acne growth and P. acne-induced inflammation. METHODS: The minerals were tested for anti-P. acne activity by disc diffusion and broth microdilution methods. The effect of these minerals on P. acne induced TNF-α and IL-8 production and gene expression were studied in THP-1 cells. In vitro toxicity was tested on human keratinocytes (HaCaT) and mouse embryonic fibroblasts (NIH3T3) using MTT assay. RESULTS: The minimum inhibitory concentrations (MIC values) for YB, TA and YBTA against P. acne were 0.1 ± 0.2, 1.9 ± 0.5 and 0.3 ± 0.5 mg mL(-1) , respectively. YB, TA and YBTA inhibited TNFα by 57.57%, 59.09% and 68.93% and IL-8 production by 48.76%, 47.92% and 51.13% in P. acne-stimulated THP-1 cells, respectively. The CTC50 values on HaCaT and NIH3T3 was 17.44 ± 0.5 and 16.37 ± 0.2 µg mL(-1) for YB, 1023.03 ± 4.0 and 1286.17 ± 4.4 µg mL(-1) for TA and 89.12 ± 2.3 and 111.58 ± 3.5 µg mL(-1) for YBTA, respectively. CONCLUSION: The present study revealed the inhibitory effect of YB, TA and YBTA on P. acne growth and inflammation. Clinical studies have suggested the anti-acne benefits of formulations containing YB and TA. The findings obtained from the present in vitro studies provide evidence to support the mechanism of anti-acne properties of YB and TA.

Ameliorative effect of Koflet formulations against pyridine-induced pharyngitis in rats.

In present study two formulations of Koflet (syrup and lozenges) were evaluated against pyridine-induced pharyngitis in rats. Topical application of 10% pyridine showed extravasation of Evans blue stain as a characteristic feature of on-going inflammation. In addition, the levels of TNF-α (p < 0.01) and IL-6 (p < 0.01) were significantly increased compared to control. Further, histopathology of the pharyngeal tissue showed submucosal gland hypertrophy, severe mucosal inflammation characterized by presence of mononuclear cells and neutrophils along with haemorrhages and congestion; however, saline applied animals (normal control) showed normal cytoarchitecture of the pharynx. Interestingly, pre-treatment with dexamethasone (1 mg/kg, p.o.), Koflet lozenges (KL) (500 and 1000 mg/kg, p.o.) and Koflet syrup (KS) (2 and 4 ml/kg, p.o.) for 7 days showed significant and dose dependent protection by decreasing the EB dye extravasation, and serum levels of TNF-α and IL-6. In addition, histopathological findings have further supported the protective effect of Koflet formulations. These findings suggest that, both Koflet syrup and Koflet lozenges are highly effective in treating non-infectious type of pharyngitis. Among the two formulations KS was found to be more potent than KL, and possible mechanism of action thought to be mediating through inhibition of TNF-α and/or phospholipids-arachidonic acid pathway.

Novel experimental model of non-infectious pharyngitis in rats.

INTRODUCTION: Currently, there is a paucity of scientific literature and reports related to screening models for non-infectious type of pharyngitis. In this context, we made a sincere attempt to establish a novel animal model for screening drugs against non-infectious pharyngitis in rats. We have considered the use of pyridine, croton oil and their combination for inducing non-infectious pharyngitis in rats. METHODS: Various concentrations of pyridine were applied topically to the pharyngeal region of rats and the extent of inflammation was assessed by Evans Blue (EB) dye exudation test, evaluating the serum levels of proinflammatory cytokines and histopathology. Dexamethasone and diclofenac were used as reference standards. RESULTS: Upon pyridine application (2.5%, 5%, 10%, 20%, 40% and 80% in saline), dose-dependent increase in EB dye extravasation was observed (increased vascular permeability). In addition, the levels of TNF-α (P<0.01) and IL-6 (P<0.01) were significantly increased compared to control. Furthermore, the histopathology of pharyngeal tissue showed hypertrophy of submucosal glands, severe inflammation of the pharynx characterised by presence of mononuclear cells, neutrophils along with haemorrhages and congestion; however, normal control animals showed normal cytoarchitecture of pharynx. Indeed, dexamethasone (0.25, 0.5 and 1 mg/kg, i.v.) and diclofenac (1, 2.5 and 5 mg/kg, i.v.) showed dose-dependent protection against pyridine-induced pharyngitis. Further, the possible mechanism of pyridine-induced pharyngitis is thought to be primarily mediated through phospholipase A2 and cyclooxygenase (COX) pathway. CONCLUSION: These findings suggest that pyridine-induced pharyngitis is a simple and versatile novel animal model for screening the drugs against non-infectious pharyngitis in rats.

Bresol inhibits phosphodiesterase 4 gene expression and modulates the levels of select mediators of inflammation in human monocytic cells.

Bresol-a poly-herbal formulation, has been reported to be effective against bronchial asthma and allergic rhinitis in children. In vivo studies have supported the anti-histaminic and anti-anaphylactic action of bresol. However, the mechanism of action of bresol in modulation of inflammation has not been studied at the cellular and molecular level. The present study was aimed to elucidate the mechanism(s) of action of bresol at the cellular and molecular levels, using human monocyte leukemia cells. The effects of bresol on phosphodiesterase 4B (PDE4B) gene expression were analyzed using human monocytic U937 leukemia cells. The ability of bresol to stimulate cAMP formation in these cells, as well as its effects on mediators of inflammation like tumor necrosis factor-α (TNFα), nitric oxide (NO), and cycloxygenase-2 (COX-2) in lipopolysaccharide (LPS)-stimulated U937 cells, were also studied. The results here indicated that bresol exhibited potential anti-inflammatory properties by inhibiting LPS-induced PDE4B gene expression in the cells. Bresol also dose dependently activated cAMP formation, and inhibited TNFα, NO, as well as COX-2 formation in the LPS-stimulated cells. Based upon the results, we concluded that the reported anti-inflammatory activity of bresol might be attributed to its abilities to inhibit PDE4B and thus elevate cAMP levels in human monocytes. The anti-inflammatory effects of bresol might also be a result of the capacity of bresol to modulate the formation of TNFα, NO, and COX-2 in monocytes.

Immunogenicity of a new, low-cost recombinant hepatitis B vaccine derived from Hansenula polymorpha in adults.

BACKGROUND: Hepatitis B virus (HBV) infection is highly prevalent world over, especially in developing countries. A new recombinant hepatitis B virus (GeneVac-B; Serum Institute of India Ltd.) vaccine is developed using Hansenula polymorpha yeast. We decided to assess the immunogenicity, and reactogenicity of this vaccine in a large adult population. MATERIAL AND METHODS: Seven hundred eighty-eight adults subjects (age: 19-57 years, male:female ratio 35:1) received three 20 microg doses of a H. polymorpha-derived recombinant hepatitis B vaccine in months 0, 1, and 6. All the eligible subjects had negative baseline serum HBs Ag, and anti-HBs. The anti-HBs titer was obtained 1 month after the last dose of vaccine and was considered seroconverted if more than 1 mIU/ml, and seroprotective if more than 10 mIU/ml. RESULTS: The seroprotection rate was 96% and seroconversion rate was 97%. Seroconversion and seroprotection rates declined with increasing age. The minimum geometric mean titre of anti HBs was 443 mIU/ml (95% CI 407-482). Seroprotection was 96% in age group<40 years, while the same was 91% in >40 years group (Odd's ratio-2.9100, Z value-2.6183, highly significant). No other factor like smoking, tobacco-chewing, alcohol consumption, chronic diseases, and obesity, affected the immune response. No significant adverse reactions were reported in any of the subjects. CONCLUSIONS: Three standard doses of the H. polymorpha-derived recombinant HBV vaccine are highly immunogenic and safe in a predominantly male adult population. Young adults respond better with this vaccine. Because of its low cost, the vaccine may be a good choice in prevention of hepatitis B infection.

Prescribing patterns among medical practitioners in Pune, India.

In 1975 the World Health Assembly requested the Director-General to advise Member States on the selection and procurement of essential drugs corresponding to their national health needs. We report here the results of a study of the prescribing patterns and rational drug utilization of medical practitioners of Pune, an industrial city in the west of India, which was undertaken by analysing their prescriptions. The results indicated a lack of rational prescribing practices by a significant number of practitioners. Fixed-dose formulations dominated the prescribing pattern and generic prescriptions were negligible, with prescriptions for essential drugs accounting for less than 60% of the total number of drugs prescribed. More than 30% of prescriptions were irrational, with the probability of such prescriptions increasing significantly with the number of drugs per prescription. A study of sources of drug formulations available for prescription revealed significantly more fixed-dose combinations, many of which were irrational. These results call for intervention strategies to promote rational drug therapy in India.

Down regulation of tumour necrosis factor activity in experimental hepatitis by a herbal formulation, Liv. 52.

A herbal hepatoprotective formulation Liv 52 down regulated the tumour necrosis factor (TNF) production in Charles Foster Rats treated with CCl4. Inhibition of TNF activity was proportional to the hepatoprotective activity.

Induction of tumour necrosis factor alpha in experimental animals treated with hepatotoxicants.

Administrations of hepatotoxicants namely carbon tetrachloride (CCl4:0.4 ml in 1.2 ml of liquid paraffin) and ANIT (1 ml of 1.5% solution in liquid paraffin) in Charles foster rats (force fed) and D-galactosamine (8 mg in water per swiss albino mouse, ip) induce the release of TNF-alpha in case of CCl4 and D-galactosamine. High TNF-alpha level was observed up to 48 hr in CCl4 and up to 24 hr in D-galactosamine treated animals. Elevated levels of biochemical like ALP and SGPT are also recorded. TNF-alpha level can be measured of tissue damage and prognosis in case of hepatitis.

Treatment of osteoarthritis with a herbomineral formulation: a double-blind, placebo-controlled, cross-over study.

The clinical efficacy of a herbomineral formulation containing roots of Withania somnifera, the stem of Boswellia serrata, rhizomes of Curcuma longa and a zinc complex (Articulin-F), was evaluated in a randomized, double-blind, placebo controlled, cross-over study in patients with osteoarthritis. After a one-month single blind run-in period, 42 patients with osteoarthritis were randomly allocated to receive either a drug treatment or a matching placebo for a period of three months. After a 15-day wash-out period the patients were transferred to the other treatment for a further period of three months. Clinical efficacy was evaluated every fortnight on the basis of severity of pain, morning stiffness, Ritchie articular index, joint score, disability score and grip strength. Other parameters like erythrocyte sedimentation rate and radiological examination were carried out on a monthly basis. Treatment with the herbomineral formulation produced a significant drop in severity of pain (P less than 0.001) and disability score (P less than 0.05). Radiological assessment, however, did not show any significant changes in both the groups. Side effects observed with this formulation did not necessitate withdrawal of treatment.

Efficacy of potassium and magnesium in essential hypertension: a double-blind, placebo controlled, crossover study.

OBJECTIVE: To evaluate the antihypertensive activity of potassium given alone or in combination with magnesium in patients with mild hypertension. DESIGN: A double blind, randomised, placebo controlled, crossover trial of 32 weeks' duration. SETTINGS: Cardiology outpatient department, Sassoon General Hospitals, Pune, India. PATIENTS: 37 Adults with mild hypertension (diastolic blood pressure less than 110 mm Hg). INTERVENTION: Patients received either placebo or potassium 60 mmol/day alone or in combination with magnesium 20 mmol/day in a crossover design. No other drug treatment was allowed. MEASUREMENTS: Blood pressure and heart rate assessed at weekly intervals and biochemical parameters at monthly intervals. RESULTS: Potassium alone or in combination with magnesium produced a significant reduction in systolic and diastolic blood pressures (p less than 0.001) and a significant reduction in serum cholesterol concentration (p less than 0.05); other biochemical variables did not change. Magnesium did not have an additional effect. Urinary potassium excretion increased significantly in the groups who received potassium alone or in combination with magnesium. The drug was well tolerated and compliance was satisfactory. CONCLUSION: Potassium 60 mmol/day lowers arterial blood pressure in patients with mild hypertension. Giving magnesium as well has no added advantage.

Clinical evaluation of terminalia belerica in diarrhoea.

Terminalia belerica (behada) fruit has been evaluated as a promising agent against E. histolytica and a variety of bacteria associated with dysentery and diarrhoea. These in vitro studies were extended further for evaluating clinical efficacy among patients of acute and chronic cases of diarrhoea and dysentery. The Studies were carried out on 25 patients by five medical practitioners practicing at different clinics in the urban areas of Pune. The data was collected as per the protocol given to the concerned clinicians. Inclusion and exclusion criteria were critically followed. Record of patients' history, clinical evaluations and investigations like stool, urine examination were carried out where necessary.The maximum treatment period was 14 days and the dose was 150 mg. tablets of bioactive fraction three times a day. 11 out of 12 patients responded to therapy and required around twelve tablets for recovery. Seven patients having the presence of cyst of amoeba, E. coli etc. became negative at the end of treatment. Improvement started on second day. No side effects of any nature were observed.

Effect of 'Arogyavardhini' against carbon tetrachloride induced hepatic damage in albino rats.

'Arogyavardhini'-an indigenous formulation was evaluated for its hepatoprotective activity in rats, using two models of carbon tetrachloride (CCl4) hepatic damage, one simulating vital hepatitis and the other simulating fatty change. The protective effect was assessed from serum aspartate transaminase (AST) and alkaline phosphatase levels and from histopathological changes in liver. The results revealed that 'Arogyavardhini' (5 mg/100g, PO daily) was effective in minimizing the changes in serum levels of AST and alkaline phosphatase induced by CCI. The protective effect was also evident on histopathological examination.