Safety and efficacy of an herbal formulation in patients with renal calculi - A 28 week, randomized, double-blind, placebo-controlled, parallel group study.

BACKGROUND: Urolithiasis is a growing problem worldwide. Many a times, asymptomatic stones are kept under observation. Many herbal preparations are available for the same, but they lack proper scientific documentation. OBJECTIVE: To study the anti-urolithiatic effect of an herbal preparation, Subap Plus (IP) capsules in patients with asymptomatic renal calculi of size ranging from 4 to 9 mm. MATERIAL AND METHODS: This was a prospective, randomized, double-blind, placebo-controlled clinical trial conducted in a tertiary care hospital in Pune, India. Patients with asymptomatic renal calculi of 4-9 mm size were randomized (1:1, block randomization) to one of the group Subap Plus (treatment group) or placebo (placebo group). The study outcome included change in visual analog scale (VAS), change in the surface area and density of calculi and their expulsion. Statistical analysis was performed using student's t-test and Chi-square test. RESULTS: A total of 120 patients were screened and 84 were enrolled who met the eligibility criteria, of which 65 patients completed the trial (treatment, n = 34; placebo, n = 31). The VAS score significantly decreased in the treatment group (6.9-1.8) than placebo group (7.2-6.8) (p < 0.001). The surface area and density were decreased by 47.58% (p < 0.008) and 43.01% (p < 0.001), respectively, in the treatment group than the placebo group. The expulsion of calculi was significantly higher in the treatment group than placebo group (20.59 vs. 3.23%, p < 0.03). CONCLUSION: Patients treated with herbal formulation showed better expulsion rate and reduction in surface area and density than the placebo group.

Studies on aerial parts of Artemisia pallens wall for phenol, flavonoid and evaluation of antioxidant activity.

Herbs have been used for medicinal purposes for centuries. According to recent investigations, they may help reduce the risk of chronic diseases, cardiovascular disease, and cancer due to antioxidant properties, which in turn can be attributed to the various phytoconstituents. With this intention, evaluation of antioxidant activity was performed. Methanol extract of aerial parts of Artemisia pallens Wall was screened for its antioxidant activity due to phenolic and flavonoid contents, by employing radical scavenging assays; 2,2 -diphenyl, 1-picryl hydrazyl (DPPH) and nitric oxide. Ascorbic acid was used as a standard. Quantitative determination of phenols and flavonoids were carried out using spectrophotometric method. Total flavonoid content was determined as quercetin equivalent and total phenolic content was determined as pyrocatechol equivalent using Folin-Ciocalteu reagent. Plant produces more phenolic compounds than flavonoids. IC(50) value of methanol extract for DPPH free radical scavenging activity was found to be 292.7 µg, whereas for nitric oxide it was 204.61 µg. The result obtained in the present study indicates that the aerial parts of this plant are a rich source of natural antioxidants.

Pre-treatment of Syndrex protects mice from becoming diabetic after streptozotocin injection.

Diabetes mellitus is a metabolic disorder characterized by hyperglycemia due to either insufficiency of insulin or inability of cells to respond to insulin. Many clinical and experimental evidence have suggested the strong association between hyperglycemia, oxidative stress and diabetic complications. Therefore, the antidiabetic drugs with antioxidant potential would have a higher therapeutic value. To check its antidiabetic and antioxidant properties in vivo, experiments were done wherein mice were fed with Syndrex in different schedules and/or made diabetic by intraperitoneal injection of streptozotocin. Animals fed with Syndrex prior to the induction of diabetes by streptozotocin injection showed resistance to an increase in blood glucose levels. This treatment increased the activities of antioxidant enzymes namely, catalase, glutathione reductase and superoxide dismutase and reduced serum triglyceride and cholesterol levels as compared to those found in uncontrolled diabetic mice. Among the three different schedules used for Syndrex treatment, the best effect was seen in the case of mice pretreated with Syndrex prior to STZ injection. In our opinion, Syndrex given along with insulin may reduce the amount of insulin dose required and because of its strong antioxidant activity would certainly help to reduce the development of diabetic complications.

CNS Depressant Activity of Ethanol Extract of Sterculia guttata seeds in mice.

Shade dried, powdered seeds of Sterculia guttata were extracted using a Soxhlet extractor with ethanol. Ethanol was removed under reduced pressure and dried to obtained crude extract. This extract was evaluated for its effect on behavioral changes, exploratory activity and barbiturate-sleeping time, using appropriate standard methods in mice. The extract exhibited dose-dependent CNS depressant activity.

Anti-inflammatory activity of Dalbergia lanceolaria bark ethanol extract in mice and rats.

Topical anti-inflammatory activity of Dalbergia lanceolaria Linn. bark ethanol extract was demonstrated in albino mice using TPA-, EPP- and AA-induced ear edema models. The systemic activity of extract was confirmed using acute and sub-acute anti-inflammatory models in albino rats. The ethanol extract exhibited significant systemic anti-inflammatory activity in Carrageenan-induced rat paw edema, by inhibition of histamine and prostaglandin phases of acute inflammation. The extract also showed significant activity against turpentine-induced exudative changes and no activity against granular tissue formation in cotton pellet-induced granuloma in albino rats. Thus, resultant anti-inflammatory activity might be due to effects on several mediators involving cyclo-oxygenase pathway resulting in prostaglandin formation and leukocyte migration from the vessels.

Antidiarrhoeal activity of Acacia nilotica Willd. bark methanol extract.

Acacia nilotica Willd. Subsp. indica (Family- Mimosaceae) bark is used for treatment of diarrhoea, dysentery, liver disorders, inflammation in ethnomedicinal practices. This bark was undertaken to study antidiarrhoeal activity using suitable battery of test. Bark powder of Acacia nilotica was successively extracted with petroleum ether, methanol and distilled water. Solvents from these extracts were removed under reduced temperature and pressure. Exploratory studies revealed presence of significant antidiarrhoeal activity only to methanol extract of Acacia nilotica (AN). This extract was studied using battery of tests consisting of castor oil and magnesium sulphate induced diarrhoea and barium chloride induced peristalsis using Swiss albino mice and in vitro antimicrobial activity against common micro-organisms causing diarrhoea. This extract showed action against castor oil, magnesium sulphate induced diarrhoea and enteropooling activity due to castor oil treatment as well as on normal as well as barium chloride induced peristalsis of small intestine significantly in mice, so also antimicrobial activity against common pathogens responsible for diarrhoea in vitro. Thus, results of above study are supporting the ethnomedicinal use of Acacia nilotica bark for the treatment of diarrhoea.

Antidiarrhoeal activity of ethanol extract of the bark of Dalbergia lanceolaria.

The bark of Dalbergia lanceolaria was undertaken for evaluation of antidiarrhoeal activity in albino mice based on ethnobotanical lead. This bark was collected from Khanapur area of Western Ghats near Pune, Maharashtra, India. The successive solvent extraction of bark was carried out after drying, using petroleum ether (60-80 degrees C) and ethanol in soxhlet apparatus. The ethanol extract showed activity against castor oil, magnesium sulphate induced diarrhoea in albino mice. The extract was also studied for castor oil induced intraluminal fluid accumulation as well as charcoal transit in normal and barium chloride treated animals. The extract reduced significantly intraluminal fluid accumulation and intestinal motility.

Protection of liver cells from ethanol cytotoxicity by curcumin in liver slice culture in vitro.

Curcumin, a widely used spice and colouring agent in food has been shown to have a broad spectrum of biological activities such as anti-inflammatory, anti-neoplastic, antimutagenic and antioxidant. We have used liver slice culture model to demonstrate hepatoprotective activity of curcumin in vitro. Ethanol has been used as a hepatotoxin and the cytotoxicity of ethanol is estimated by quantitating the release of LDH. Ethanol induces 3.5 times more release of LDH from the liver cells and twice the amount of lipid peroxidation as compared to the cells from untreated liver tissue and this was significantly reduced in presence of curcumin (5 microM). We measured the activity of antioxidant enzymes (AOEs) namely superoxide dismutase, catalase and peroxidase and found that in ethanol treated cells activity of all three enzymes was elevated. However, when curcumin was added along with ethanol their levels were kept low. The fact that release of LDH is significantly reduced along with lipid peroxidation and the activity of AOEs is kept low indicates that curcumin by its antioxidant activity reduced the oxidative stress induced by ethanol and protected the liver cells in vitro.

Taraxer-14-en-3beta-ol, an anti-inflammatory compound from Sterculia foetida L.

Taraxer-14-en-3beta-ol ( 1) was shown to be the active ingredient in the leaves of Sterculia foetida L. The alcohol 1, its acetate and ketone showed anti-inflammatory activity against TPA induced mouse ear oedema with inhibition ratios of 60.0, 58.57 and 40.57 at 0.5 mg/ear, respectively. The percentage inhibition of inflammation increased with dose for each compound.

Anti-inflammatory activity of Jatropha curcas roots in mice and rats.

Anti-inflammatory activity of topical application of Jatropha curcas L. root powder in paste form in TPA-induced ear inflammation was confirmed in albino mice and the successive solvent extraction of these roots was carried out by ether and methanol. The methanol extract exhibited systemic and significant anti-inflammatory activity in acute carrageenan-induced rat paw edema. It also showed activity against formalin-induced rat paw edema, as well as, turpentine-induced exudative changes and cotton pellet-induced granular tissue formation after oral treatment for 7 days in albino rats. Thus, resultant anti-inflammatory activity might be due to effects on several mediators and arachidonic acid metabolism involving cyclo-oxygenase pathway resulting in prostaglandin formation, anti-proliferative activity leading to reduction in granular tissue formation and leukocyte migration from the vessels.

Isolation and X-ray study of an anti-inflammatory active androstene steroid from Acacia nilotica.

3beta-Acetoxy-17beta-hydroxy-androst-5-ene was isolated from aerial parts of Acacia nilotica (L.) Willd (Mimosaceae). The structure of this compound was established by spectral analysis and single crystal X-ray diffraction analysis. The steroid showed dose-dependent anti-inflammatory activity against TPA-induced mouse earedema.

Reproductive toxicity of piperine in Swiss albino mice.

Piperine (CAS 94-62-2) is a constituent of various spices which are used as common food additives all over the world. The reproductive toxicity of piperine was studied in Swiss albino mice. Relevant short-term tests were employed to assess the effect on estrous cycle, mating behaviour, toxicity to male germ cells, fertilization, implantation and growth of pups. Piperine (10 and 20 mg/kg b.w.) increased the period of the diestrous phase which seemed to result in decreased mating performance and fertility. Post-partum litter growth was not affected by the piperine treatment. Sperm shape abnormalities were not induced by piperine at doses up to 75 mg/kg b.w. Considerable anti-implantation activity was recorded after five days post-mating oral treatment with piperine. The sex ratio and post-implantation loss were unaffected after treatment with piperine. Intrauterine injection of piperine caused the total absence of implants in either of the uterine horns (16.66%) or one of the horns (33%) of treated females. No histopathological changes were detected in the ovary and the uterus at the cellular level. Prostaglandin E1-induced acute inflammation of rat paw was significantly reduced after piperine treatment. Our results show that piperine interferes with several crucial reproductive events in a mammalian model.

Studies on antidiarrhoeal activity of Jatropha curcus root extract in albino mice.

Use of Jatropha curcus L. roots in the treatment of diarrhoea is a common ethnobotanical practice in Konkan, a part of the Western coastal area of India. Roots of this species were undertaken for pharmacognostic studies and evaluation of antidiarrhoeal activity in albino mice. Successive solvent extraction was carried out using petroleum ether (60-80 degrees C) and methanol. The methanol extract showed activity against castor oil induced diarrhoea and intraluminal accumulation of fluid. It also reduced gastrointestinal motility after charcoal meal administration in albino mice. The results indicate that action of J. curcus root methanol extract could be through a combination of inhibition of elevated prostaglandin biosynthesis and reduced propulsive movement of the small intestine.

Pharmacological studies on sterculia foetida leaves.

An alcoholic extract of Sterculia foetida L. leaves was subjected to pharmacological screening using various animal models. The extract caused reduced exploratory activity in mice. Further it potentiated pentobarbitone sleeping time in normal and chronic pentobarbitone-treated mice. It also potentiated barbital sodium-induced hypnosis, indicating central nervous system depressant activity. The extract also exhibited significant antiinflammatory activity in the acute carrageenan-induced rat paw edema and the chronic granuloma pouch models. However, it was devoid of analgesic activity in the tail flick model.

Assessment of genotoxic effect of piperine using Salmonella typhimurium and somatic and somatic and germ cells of Swiss albino mice.

Piperine (CAS 94-62-2) is a constituent of various spices and is used as a common food additive all over the world. The genotoxic potential of piperine was assessed using four different test systems, namely, Ames test using Salmonella typhimurium, micronucleus test, sperm shape abnormality test and dominant lethal test using Swiss albino mice. In the Ames test, six different doses of piperine, in the range of 0.005-10 mumol/plate, did not induce his+ revertants, with or without metabolic activation, indicating its nonmutagenic nature. In the bone narrow micronucleus test using two doses in the range of therapeutic usage (10 and 20 mg/kg body weight), piperine itself was non-mutagenic. Like in somatic cells, piperine (10 and 50 mg/kg body weight) failed to induce mutations in male germ cells of mouse as assessed by using the sperm shape abnormality and dominant lethal tests. Piperine thus appears to be a non-genotoxic chemical.

Toxicological profile of aureofungin: a potent plant antifungal antibiotic.

Aureofungin is a heptaene type of antifungal antibiotic used for controlling plant fungal infections and diseases, during pre and post harvesting period of various crops. Acute and subacute oral toxicity of aureofungin in rats was studied along with haematological, urine analysis and other biochemical parameters related to liver and kidney organ functions. The results of these studies indicate mild toxic symptoms at higher doses which were reversible following its withdrawal.

A comparative pharmacological investigation of Ashwagandha and Ginseng.

The aqueous suspensions of roots of an Indian drug Ashwagandha and the Korean drug Ginseng were tested comparatively for 2 pharmacological activities, namely the anti-stress activity by the 'mice swimming endurance test' and anabolic activity by noting gain in body weights and levator ani muscle in rats. A significant increase in mice swimming time was shown by Ginseng (P < 0.001) and Ashwagandha (P < 0.01) as compared to the control group. Significant increase in body weights in the Ashwagandha treated group (P < 0.05) was better than Ginseng (P < 0.5). Gain in wet weights of the levator ani muscle were also significant in Ginseng (P < 0.001) and Ashwagandha (P < 0.01) treated groups, however, the weight gain of dried levator ani muscles showed comparable results for both these drugs (P < 0.01).

Genotoxicity assessment of the antifungal antibiotic aureofungin in Salmonella typhimurium and Swiss albino mice.

The widely used agricultural antifungal agent aureofungin (ARF) was subjected to genotoxicity assessment using the Ames Salmonella assay as well as the in vivo micronucleus test and dominant lethal test in Swiss mice. In the Ames Salmonella spot test, ARF slightly elevated the number of histidine revertants after metabolic activation over a wide dose range (1-1000 micrograms/plate) in TA102 but not in TA97a, TA98 or TA100. In the preincubation plate incorporation assay with TA102, ARF increased the number of revertants in a dose-dependent manner only after metabolic activation. ARF failed to significantly elevate the frequency of micronucleated polychromatic erythrocytes (PE) in the bone marrow of Swiss mice. It elevated the frequency of dominant lethal mutations in the 7th and 8th weeks at 30 mg/kg body weight, a concentration much higher than the actual concentration used in the field. We conclude that ARF is non-mutagenic in somatic cells in vivo at doses used in the present study, probably mutagenic in stem-cell spermatogonia and may be classified as an equivocal promutagen, possibly acting as a cross-linker.

Inhibition of lipid peroxidation by piperine during experimental inflammation in rats.

Carrageenin induced rat paw oedema shows a direct co-relationship with liver lipid peroxidation and not with kidney or brain. Pretreatment with piperine or oxyphenylbutazone reduced the liver lipid peroxidation, acid phosphatase and oedema induced by carrageenin. However, no such co-relationship was observed with treatment of these anti-inflammatory agents in control animals. It is, therefore, suggested that the inhibition of these liver enzymes is non specific in nature.

Anti-inflammatory activity of piperine.

Piperine (1-peperoyl piperidine) was isolated from Piper nigrum Linn for the evaluation of anti-inflammatory activity in rats. Different acute and chronic experimental models like carrageenin-induced rat paw edema, cotton pellet granuloma, and croton oil-induced granuloma pouch, were employed. Simultaneously, biochemical estimations were made to elucidate the underlying mechanism of the action. Piperine acted significantly on early acute changes in inflammatory processes and chronic granulative changes. It also acted partially through stimulation of pituitary adrenal axis. Exudative changes in both acute and chronic models, however, were insignificant.