Effect of Ayurvedic mercury preparation Makaradhwaja on geriatric canine--a preliminary study.

Makaradhwaja, an alchemical Ayurvedic mercury preparation is used as stimulant and vitalizer. Towards veterinary practices, the acceptability, tolerability and toxicity studies were undertaken in geriatric pet dogs aged more than 10 years irrespective of breed and sex for future use. Makaradhwaja (2.5 mg/kg) was used with honey once daily for 30 days. Before and after treatment, blood was collected for hematological studies as well as liver, kidney function and anti-oxidant activity. In control group, honey itself showed no appreciable change whereas, Makaradhwaja lowered neutrophil and total leucocyte count. Serum cholesterol, urea, glucose, alanine amino transferase, aspartate amino transferase, sodium, phosphorus and calcium were decreased. Haemoglobin and serum creatinine were significantly increased. There was appreciable physical, behavioral and body weight change including quality of life. The dose was used in replication of human dose (125 mg/50 kg). Anti-oxidant study showed significant increase of lipid per oxidation in experimental group while the values of ABTS radical cation decolorisation assay although decreased but did not show any significant changes. Decrease of serum urea and increase of serum creatinine could not be explained on single dose response. Different dose study could only explain the optimum dose to be required in canine practices.

Clinical evaluation of spermatogenic activity of processed Shilajit in oligospermia.

The safety and spermatogenic activity of processed Shilajit (PS) were evaluated in oligospermic patients. Initially, 60 infertile male patients were assessed and those having total sperm counts below 20 million ml(-1) semen were considered oligospermic and enrolled in the study (n = 35). PS capsule (100 mg) was administered twice daily after major meals for 90 days. Total semenogram and serum testosterone, luteinising hormone and follicle-stimulating hormone were estimated before and at the end of the treatment. Malondialdehyde (MDA), a marker for oxidative stress, content of semen and biochemical parameters for safety were also evaluated. Twenty-eight patients who completed the treatment showed significant (P < 0.001) improvement in spermia (+37.6%), total sperm count (+61.4%), motility (12.4-17.4% after different time intervals), normal sperm count (+18.9%) with concomitant decrease in pus and epithelial cell count compared with baseline value. Significant decrease of semen MDA content (-18.7%) was observed. Moreover, serum testosterone (+23.5%; P < 0.001) and FSH (+9.4%; P < 0.05) levels significantly increased. HPLC chromatogram revealed inclusion of PS constituents in semen. Unaltered hepatic and renal profiles of patients indicated that PS was safe at the given dose. The present findings provide further evidence of the spermatogenic nature of Shilajit, as attributed in Ayurvedic medicine, particularly when administered as PS.

Screening of antioxidant activity of three Indian medicinal plants, traditionally used for the management of neurodegenerative diseases.

A number of Indian medicinal plants have been used for thousands of years in the traditional system of medicine (Ayurveda). Amongst these are plants used for the management of neurodegenerative diseases such as Parkinson's, Alzheimer's, loss of memory, degeneration of nerves and other neuronal disorders by the Ayurvedic practitioners. Though the etiology of neurodegenerative diseases remains enigmatic, there is evidence, which indicates that defective energy metabolism, excitotoxicity and oxidative damage may be crucial factors (Ann. Neurol. 38 (3) (1995) 357). The part of the Ayurvedic system that provides an approach to prevention and treatment of degenerative diseases is known as Rasayana, and plants used for this purpose are classed as rejuvenators. This group of plants generally possesses strong antioxidant activity (Pharmacol. Biochem. Behav. 43 (1992) 1175), but only a few have been investigated in detail. In the present study, three such rasayana plants were tested for the first time for their toxicity and free radical scavenging activity both in vitro and ex vivo. All the three plant infusions (up to 1 mg/ml) showed no toxic effects on the viability of PC12 cell line as judged by MTT-test. Both ethanolic extracts and water infusions of the plants were tested for their antioxidant activity in the 2,2'-azinobis-3-ethyl-benzothiazoline-6-sulfonic acid (ABTS*(+)) radical cation decolorization assay; inhibition of lipid peroxidation by plant infusions was carried out using spontaneous lipid peroxidation of rat brain homogenate, and IC50 values were determined. The results from the ABTS assay showed that the ethanolic extract of Sida cordifolia was found to be most potent (IC50 16.07 microg/ml), followed by Evolvulus alsinoides (IC50 33.39 microg/ml) and Cynodon dactylon (IC50 78.62 microg/ml). The relative antioxidant capacity for the water infusions was observed in the following order: E. alsinoides (IC50 172.25 microg/ml)>C. dactylon (IC50 273.64 microg/ml)>S. cordifolia (IC50 342.82 microg/ml). The results of water infusions of the plants on lipid peroxidation were as follows: E. alsinoides (IC50 89.23 microg/ml)>S. cordifolia) (IC50 126.78 microg/ml)>C. dactylon (IC50 608.31 microg/ml).

Evaluation of chemical constituents and free-radical scavenging activity of Swarnabhasma (gold ash), an ayurvedic drug.

From ancient times, Swarnabhasma (gold ash) has been used in several clinical manifestations including loss of memory, defective eyesight, infertility, overall body weakness and incidence of early aging. Swarnabhasma has been used by Ayurvedic physicians to treat different diseases like bronchial asthma, rheumatoid arthritis, diabetes mellitus, nervous disorders, etc. In the present investigation, Swarnabhasma was prepared after proper purification and calcination as per Ayurvedic pharmacy which consisted of Realger (As(2)S(2)), Lead oxide (Pb(3)O(4)), Pure gold (Au) and Latex of Calotropis gigantea. Qualitative analyses indicated that Swarnabhasma contained not only gold but also several microelements (Fe, Al, Cu, Zn, Co, Mg, Ca, As, Pb, etc.). Infrared spectroscopy showed that the material was free from any organic compound. The metal content in the bhasma was determined by atomic absorption spectrometry. Acute oral administration of Swarnabhasma showed no mortality in mice (up to 1 ml /20 g b.w. of Swarnabhasma suspension containing 1mg of drug). Chronic administration of Swarnabhasma also showed no toxicity as judged by SGPT, SGOT, serum creatinine and serum urea level and histological studies. In an experimental animal model, chronic Swarnabhasma-treated animals showed significantly increased superoxide dismutase and catalase activity, two enzymes that reduce free radical concentrations in the body.

Wound healing activity of human placental extracts in rats.

AIM: To study wound healing activity of the human placental extract (HPE) in rats. METHODS: Full thickness wounds were inflicted on depilated dorsum of Charles foster rats with 8 mm Acu-punch biopsy. The HPE was applied both at topical and im routes (2.5 mL/kg). Effects were compared on the basis of physical criteria, biochemical criteria, and histopathological study with respect to untreated control, vehicle control (1.5 % benzyl alcohol), and framycetin topical treated groups. RESULTS: Significant lowering of wound size (P<0.05), wound index (P<0.05), and number of days required for complete healing (P<0.01); significant gain in tensile strength (P<0.01); appreciable increase of tissue DNA, total protein, and collagenesis were observed in HPE treated group. CONCLUSION: Human placental extract systematically helps collagenesis leading to potent healing of wounds.

A lethal cardiotoxic protein isolated from Bidder's organ of common Indian toad, Bufo melanostictus Schneider.

A lethal cardiotoxic (BO-CT; Bidder's organ cardiotoxin) protein was purified from the Bidder's organ of the common Indian toad B. melanostictus by gel filtration on Sephadex G-200. The homogeneity of cardiotoxin was tested by gel electrophoresis. The molecular weight of lethal BO-CT was 62 KDa and was devoid of glycoprotein. LD50 of the BO-CT was 50 micrograms/20 g (i.v.) in male albino mice. On isolated heart and auricle BO-CT initially increased the rate and amplitude of contraction and finally produced irreversible blockade of contraction. BO-CT induced auricular blockade, was not influenced by verapamil, propranolol and atropine. On isolated chick biventer cervicis preparation BO-CT produced irreversible blockade of electrically induced twitch response followed by contracture. This action was not antagonized by 4-aminopyridine and neostigmine. BO-CT induced contracture on chick biventer cervicis was increased by Ca2+, decreased by Na+ and abolished by K+. Cardiotoxic and neuromuscular activity of BO-CT was heat stable and abolished by proteolytic enzyme.

A lethal protein toxin (toxin-PC) from the Indian catfish (Plotosus canius, Hamilton) venom.

Isolation and purification of a lethal protein toxin from the Indian catfish Plotosus canius, Hamilton, venom is described. The purification procedure involved ammonium sulfate precipitate of crude venom followed by DEAE-ion exchange chromatography. The purified toxin (toxin-PC) was homogeneous on one-dimensional PAGE and PAS-negative, and had a molecular weight 15 Kd. Toxin-PC was lethal (LD50 225 micrograms/kg, intravenous, in mice) and cardiotoxic, having neuromuscular blocking activity. Toxin-PC produced cardiac arrest on isolated toad and guinea pig hearts. Prior administration of atropine and propanolol failed to counteract toxin activity on isolated heart preparations. On isolated chick biventer cervicis, toxin-PC produced total blockage of electrically-induced twitch response without affecting carbachol- and acetylcholine-induced contraction. The tension developed by the muscle was Ca++ ion-dependent. Neuromuscular blocking time was reduced when K+ ion concentration was increased in the medium. Antiserum raised against toxin-PC failed to antagonize lethal activity of toxin-PC in mice. Toxin-PC probably represents a major toxic component of catfish venom (P. canius), and was responsible for the pathophysiological changes.

Isolation, purification and partial characterization of viper venom inhibiting factor from the root extract of the Indian medicinal plant sarsaparilla (Hemidesmus indicus R. Br.).

An organic acid, isolated and purified from the root extract of an Indian medicinal plant sarsaparilla Hemidesmus indicus R. Br, possessed viper venom inhibitory activity. The compound (designated HI-RVIF) was isolated by solvent extraction, silica gel column chromatography and thin layer chromatography, and was homogeneous in nature. The white needle-shaped crystals were soluble in water, methanol and chloroform and had a melting point of 155-158 degrees C and lambda max 260 nm. Spectral analysis confirmed the presence of a benzene ring, methoxy group, and hydroxyl group; the mol. wt of the compound was 168. HI-RVIF significantly antagonized viper venom-induced lethal, haemorrhagic, coagulant and anticoagulant activity in experimental rodents.

Isolation, purification and partial characterization of a haemolytic protein from Bidder's organ of toad Bufo melanostictus (Schneider).

A haemolytic protein toxin (BO-HT) from Bidder's organ of toad, B. melanostictus, purified by DEAE-cellulose column chromatography was electrophoretically homogeneous and was glycoprotein in nature (PAS-positive). The molecular weight was estimated to be 14.4 kDa by SDS-polyacrylamide gel electrophoresis. The sensitivity of the haemolysin of different RBC ghost cell preparation was in the order: buffalo > goat > ox > guinea pig > mice > human > chick > rabbit > rat. The haemolytic activity was increased with the decrease in RBC concentration and was produced over a wide range of temperature. Maximum haemolytic effect was produced at 2 hr of incubation. The toxin showed maximum activity at 3 and minimum at 10 pH. Divalent cations (Ca2+, Zn2+, Cu2+, Mg2+) showed inhibitory effect on BO-HT induced haemolysis, whereas sucrose, EDTA, cholesterol, 2-mercaptoethanol and oxygen did not alter the haemolytic activity. Haemolytic activity was reduced by proteolytic enzymes (trypsin, protease) and was totally antagonized by the toad serum.

Pharmacological actions of the venom of the Indian catfish (Plotosus canius Hamilton).

The venom of the common Indian catfish P. canius Hamilton (locally called 'Kanmagur') was examined for its pharmacodynamic activity. The LD50 of the venom in mice was found to be 3.9 mg/kg (ip). At lower doses, the venom produced a positive inotropic effect on toad and rabbit hearts, while at higher doses it produced cardiac arrest. In the isolated guinea pig auricle, the venom increased the rate and amplitude of contraction. The venom increased rat blood pressure--an action antagonised by alpha-adrenergic blocker (phenoxybenzamine). It reduced the rate and amplitude of rat and guinea pig respiration leading to respiratory arrest and death. The venom did not alter the cutaneous capillary permeability of guinea pig but produced vasoconstrictor effect on rat hindquarter perfusion. It induced contractions in several smooth muscle preparations viz., ileum and colon of guinea pig, fundus, uterus and ileum of rat. On isolated guinea pig ileum, the venom produced contraction which was not antagonised by atropine and mepyramine, but was partially antagonised by methysergide associated with a residual contraction which was abolished by SC 19220, a prostaglandin receptor blocker. The venom produced irreversible blockade of electrically induced twitch response on isolated rat phrenic nerve diaphragm and chick biventer cervicis preparation. Haemolysis was not produced by the venom on mice, guinea pig and human RBC (washed).