Polyherbal formulations with wide spectrum antimicrobial activity against reproductive tract infections and sexually transmitted pathogens.

PROBLEM: Recent reports indicate high incidence of genital infections, most of which are sexually transmitted. Although specific drugs and antibiotics are available for some, a safe spermicidal formulation with wide spectrum antimicrobial action would be a desirable addition to the presently available spermicides. METHODS: Formulations at different dilutions were tested in culture systems on standard strains and clinical isolates including some isolates resistant to drugs. The effect on (HSV)-2 and Chlamydia trachomatis was determined in vivo in progestin sensitized mice. The effect on HIV-1 was investigated in two standardized systems. RESULTS: Polyherbal cream inhibited the growth in culture of clinical isolates of Candida albicans, Candida krusei and Candida tropicalis. Both the polyherbal cream and the Praneem polyherbal pessary inhibited urinary tract Escherichia coli (including multidrug resistant strains), and Neisseria gonorrhoeae (including 2 strains resistant to penicillin). Both formulations manifested virucidal activity against HIV-1 at >2 and 50% dilutions (in two different test systems) on contact for 1-2 min. Intravaginal inoculation of the cream and the pessary suspensions before inoculation of the pathogen prevented lesions and vaginal transmission of HSV-2 and C. trachomatis in progestin sensitized mice. CONCLUSIONS: Polyherbal formulations have wide spectrum antibacterial, antifungal and antiviral effect against the tested sexually transmitted pathogens.

Broad spectrum herbal therapy against superficial fungal infections.

Skin disease associated with keratinized tissues in animal and human beings has been investigated. The essential oil of Eucalyptus pauciflora in vitro showed strong antifungal activity at 1.0 microl/ml against human pathogenic fungi, viz. Epidermophyton floccosum, Microsporum audouinii, M. canis, M. gypseum, M. nanum, Trichophyton mentagrophytes, T. rubrum, T. tonsurans and T. violaceum. The oil has heavy doses of inoculum potential at 1.0 microl/ml. Moreover, it did not exhibit any adverse effects on mammalian skin up to 5% concentrations. Further, we formulated the oil in the form of ointment 'BSHT' (broad spectrum herbal therapy) (1% v/v) and subjected it to topical testing on patients attending the outpatient department of M.L.N. Medical College, Allahabad. Fifty patients were selected on the basis of KOH-positive results and diagnosed as either tinea pedis, tinea corporis or tinea cruris. After the second week of treatment, all patients were KOH-negative. At the end of medication, 60% of patients recovered completely and 40% showed significant improvement from the disease. No KOH-negative cases of relapse were observed when patients were re-examined after 2 months following the end of treatment. Thus, the ointment can be exploited commercially after undergoing successful multicenter clinical trials, which are in progress.

Progression of mouse buthionine sulfoximine cataracts in vitro is inhibited by thiols or ascorbate.

Mouse lens cultures were employed to study the progression of cataracts initiated by injection of buthionine sulfoximine, an inhibitor of glutathione (GSH) biosynthesis. Culture of lenses removed from untreated mice on postnatal day 7, for 48 hr in the presence of 4 mm BSO, resulted in only limited cataractous changes. To enable substantial progression of cataracts in vitro, it was therefore necessary to pretreat the mice with BSO prior to lens culture. A single injection of BSO (4 nmol mg-1 lens), administered on day 7, resulted in >90% depletion of lens GSH within 3 days, but no visible cataractous changes. The clear lenses were incubated for 29+/-1 hr at 37 degrees C in Medium HL-1, supplemented with EGF, insulin and Ca2+, in the presence or absence of BSO, and were scored for cataract development by previously described criteria. In the absence of BSO, only 4 of 10 lenses developed large opacities. However, in the presence of 4 mm BSO, 40 out of 45 experimental lenses developed opacities affecting at least 50% of the lens visual field and were scored as stages 1C-4, depending upon the extent and density of the cataracts. In addition, three lenses had opacities involving 20-50% of the field (stage 1B). By contrast, less than 10% of lenses from untreated mice incubated in the absence of BSO developed opacities. The cataracts developed in 4 mm BSO were accompanied by reduction of lens glutathione levels to <0.010 nmol mg-1 lens. They were almost completely prevented by 1 mm ascorbate, 2 mm GSH, 2 mm GSH monoethyl ester and 2 mm cysteamine. GSH and GSH ester maintained lens glutathione content between 0.1 and 0.2 nmol mg-1 in the presence of BSO, whereas ascorbate did not prevent near-total GSH depletion. The prevention of cataracts by thiols and ascorbate was confirmed by lens Na/K ratios not significantly different from those in control lenses. The above combination of GSH depletion in vivo by a single injection of BSO, followed 3 days later with lens culture in the presence of BSO, may yield a useful system to elucidate and control the biochemical mechanisms involved in oxidative cataract induction by this GSH-depleting agent.

Effects of certain atmospheric pollutants (SO2, NO2 and CO) on the soluble amino acids, molecular weight and antigenicity of some airborne pollen grains.

The pure pollen grains of Red Oak (Quercus rubra), Meadow Fescue (Festuca elatior) and Chinese Elm (Ulmas pumila) were exposed to carbon monoxide (CO), sulphur dioxide (SO2) and nitrogen dioxide (NO2). After exposure, the soluble free amino acids were determined from the extracts using two-dimensional thin layer chromatography, and the molecular weight of the extracts were determined by SDS-gel electrophoresis (PAGE). The results indicated that after contamination, both the amino acids and molecular weight profiles were changed. In addition, the double immunodiffusion method was used against rabbit-antisera to determine the antigenicity of contaminated and non-contaminated pollen grain extracts. The results also showed that there were antigenic changes after contamination.

Garlic as a natural agent for the treatment of hypertension: a preliminary report.

The major objective of this study was to re-evaluate the effects of garlic on blood pressure with respect to its ability to provoke a decrease in blood pressure and to determine the length of time that this decrease would require. Spontaneously hypertensive rats were given three doses of garlic extract (0.1 ml/kg, 0.25 ml/kg, and 0.5 ml/kg) by oral injection. The blood pressures of these ether-anaesthetized rats were measured immediately before the extract was given, and then 0.5, 2, 4, 6, and 24 h after the extract was given. A blood pressure measurement was also taken at 48 h after extract administration for the 0.5 ml/kg dose. The Gilson Duograph System was used to measure blood pressure by the tail-cuff method. There was a marked decrease in the systolic blood pressure of all of the rats after three doses and the decrease occurred within 30 min in each case. Even though the average decreases for the 0.1 ml/kg and the 0.25 ml/kg doses were calculated as 51,25 mm Hg and 56.25 mm Hg, respectively, these doses were not sufficient to sustain the blood pressure in a normal range for more than 1 or 2 h. The 0.5 ml/kg dose, showing an average decrease of 65.7 mm Hg, was sufficient to provoke a decrease to a normal level and to sustain this decrease for up to 24 h. The results indicate that garlic is effective as a natural agent for the treatment of hypertension.

Antistress and antifatigue properties of Panax ginseng: comparison with piracetam.

The antistress and antifatigue properties of a Chinese ginseng preparation were tested on Swiss albino mice, exposed to various experimental models of stress, and were compared with those of piracetam. Both ginseng and piracetam were administered chronically in drinking water for 16-18 days as well as acutely, by injection, 30-60 min prior to the experiments. Reactivity of the mice, loss in body weight, amount of faeces, length of endurance and incidence of mortality were graded and measured. Both piracetam and ginseng treatment provided good protection against electroshock stress when compared to the untreated mice; fighting scores, incidence of tonic convulsion and mortality were significantly less in the treated groups. In the heat stress experiments, both piracetam and ginseng provided significant protection to the treated mice against exposure to heat. In the fatigue stress of forced swim test, ginseng treatment provided effective adaptation to fatigue and increased endurance in both male and female mice; piracetam showed some antifatigue effects on the male mice only. In the locomotor activity tests, ginseng did not depress motility, while piracetam did so in the later part of the tests. These results are discussed in the light of the antistress properties of the drugs as reported in the literature.

The temporal dimensions of anticonvulsant action of some newer benzodiazepines against metrazol induced seizures in mice and rats.

In a time-distribution study, the anticonvulsant effects of four benzodiazepine compounds were compared with those of three standard antiepileptics against metrazol-induced seizures in mice and rats. Ethosuximide and trimethadione had the shortest duration of action in mice, but protected the rats up to 6 hr. Phenobarbitone, diazepam, flurazepam and nitrazepam protected the mice up to 12 hr, but the rats were effectively protected only up to 3-4 hr. Clonazepam, the most potent and effective agent, protected the mice from clonic-tonic seizures up to 18-20 hr and the rats up to 6-7 hr. Comparison of the PD50 from clonic seizure at the peak-effect hours revealed that the benzodiazepines were 16 to 96 times more potent than phenobarbitone on a molar basis, while phenobarbitone itself was 12 to 26 times more potent than ethosuximide and trimethadione. Tonic seizures and mortality were largely suppressed by all drugs until 18-20 hr in mice and 6-7 hr in rats. Seizure latency and mortality patterns varied from drug to drug but not in a dose-dependent manner.

Microinjections of tubocurarine, leptazol, strychnine and picrotoxin into the cerebral cortex of anaesthetized cats.

1. In cats anaesthetized with intravenous chloralose, microinjections of tubocurarine, leptazol, strychnine or picrotoxin, in a volume of 1 mul, were made into the grey matter of the cerebral cortex and the electrical activity was recorded from the site of injection with the microinjection cannula which, insulated except at its tip, served as recording electrode.2. Routinely the injections were made into the gyrus splenialis or into the underlying gyrus cinguli close to the mid-line, because the injections would then most likely be in grey and not in white matter. Injected in this way all four drugs set up foci of excitation which gave rise to synchronous firing of a large number of neurones with the result that high voltage negative spikes were recorded from the microinjection cannula.3. On injection into the gyrus splenialis the threshold dose was about 0.04 mug for picrotoxin, about 0.2 mug for tubocurarine, about 5 mug for strychnine and 25 to 50 mug for leptazol. Following the injection of larger doses the spike discharge continued for a few hours after picrotoxin and tubocurarine, for over an hour after strychnine, but for a few minutes only after leptazol. On injection into the gyrus cinguli the threshold doses were slightly greater and with larger doses the spikes occurred at greater frequency but were of lower voltage than in the gyrus splenialis.4. With large doses of picrotoxin injected into the gyrus splenialis the spikes developed an after-positivity and an after-discharge which sometimes passed into a short period of fast activity.5. The foci of excitation set up by the drugs were restricted to the site of injection because on raising or lowering the microinjection cannula the spikes recorded from it quickly decreased in voltage and then disappeared. When the injections were made close to a sulcus and the microinjection cannula, on being lowered, traversed the sulcus, the spikes changed their polarity.6. The spike discharge appears to be a consistent response to the injections of the drugs into grey matter of any part of the cerebral cortex since it was also obtained on their injection into the pyriform cortex, amygdala and area retrolimbica anterior, but not on their injection into white matter or caudate nucleus, thalamus or hypothalamus.

Temperature effects of reserpine injected into the cerebral ventricles of rabbits and cats.

1. In unanaesthetized rabbits and cats reserpine was injected through a chronically implanted cannula in the left lateral cerebral ventricle, and rectal temperature was recorded.2. In rabbits the reserpine (0.5-0.6 mg) caused a rise in temperature, frequent defaecation and sedation. On repeating the intraventricular injections at 24 hr intervals the rise in temperature was not obtained with the second or third injection, but defaecation and sedation still occurred. When the hyperthermic response to intraventricular reserpine had disappeared the anterior hypothalamus still responded to intraventricular noradrenaline which produced a rise in temperature.3. In cats the reserpine (0.5-0.75 mg) caused a biphasic change in temperature, i.e. an initial fall followed by a rise, frequent defaecation, and catalepsy. On repeating the intraventricular injections at 24 hr intervals the initial hypothermic phase of the temperature response was not obtained with the second or third injection, but the late rise, defaecation and catalepsy were still produced. When the hypothermic phase had disappeared the hypothalamus still responded to intraventricular noradrenaline or adrenaline which produced a fall, and to intraventricular 5-hydroxytryptamine (5-HT) which produced a rise in temperature.4. It is concluded that the rise in temperature in rabbits and the initial fall produced in cats is not due to a direct action of reserpine on the cells of the anterior hypothalamus but to noradrenaline released from adrenergic fibres ending at these cells. When these fibres are depleted of their noradrenaline by one or two injections of reserpine, these effects are not obtained because noradrenaline is no longer available to be released in sufficient amounts to raise temperature in rabbits and to lower it in cats.

Effect on temperature of 5-hydroxytryptamine injected into the cerebral ventricles of cats.

1. In unanaesthetized cats the effect on rectal temperature was examined of 5-HT injected through a Collison cannula chronically implanted into the left lateral ventricle. The response depended on the amount of 5-HT injected and on the solvent employed.2. An intraventricular injection of 200 mug 5-HT creatinine sulphate dissolved in 0.9% NaCl solution resulted in a long-lasting rise often interrupted initially by a transient fall in temperature.3. This fall became more prominent with larger doses of 5-HT; even more when the 5-HT was dissolved in distilled water, and then the hyperthermic effect was attenuated.4. It is concluded that intraventricular 5-HT raises rectal temperature in cats when the amount is not too large, and that a hypothermic effect when it occurs results from paralysis of cells in the anterior hypothalamus which are excited by small doses. This would be similar to the actions of acetylcholine in the perfused superior cervical ganglion of the cat, where small doses excite but large doses paralyse the ganglion cells.5. An intraventricular injection of distilled water produced a steady rise in temperature which is attributed to release of 5-HT.