Simple liquid chromatography-tandem mass spectrometry method for determination of novel anti-methicillin-resistant Staphylococcus aureus fluoroquinolone WCK 771 in human serum.

A simple, rapid, specific, precise, accurate and sensitive method for determination of WCK 771 in human serum has been developed. The method uses high performance liquid chromatography with tandem mass spectrometric detection. Sample preparation involves protein precipitation method by addition of acetonitrile. Gatifloxacin was used as internal standard. The response was found to be linear from 0.312 to 40 microg/ml of serum with correlation coefficient greater than 0.99. Limit of detection and lower limit of quantification for WCK 771 was found to be 0.078 microg/ml and 0.312 microg/ml, respectively. The intra-day precision and accuracy from analysis of quality control (QC) samples at four concentrations was in the range of 2.36-2.58% and from 96.71 to 103.2%, respectively. The inter-day precision and accuracy from analysis of quality control samples at four concentrations was in the range of 3.14-6.82% and from 96.84 to 105.76%, respectively. WCK 771 was found to be stable for 24 h at auto-injector environment. WCK 771 was also found to be stable for 2h in serum at 25+/-3 degrees C and for 3 months at -20 degrees C. Mean absolute recovery at four different concentrations was 86.92% with standard deviation of 1.79. Throughput of the method is approximately one sample every 4 min. The method was also reproduced with monkey serum. The method was employed for estimation of drug serum levels during pre-clinical and clinical trials.

The C of CHADS: Historical perspective and clinical applications for anticoagulation in patients with non valvular atrial fibrillation and congestive heart failure.

The risk stratification of patients with coexisting non valvular atrial fibrillation and congestive heart failure, is often a clinical challenge, as the definitions of congestive heart failure in the popular CHADS2 and CHA2DS2VASc scoring systems, and amongst major clinical trials on Warfarin and Novel Oral Anticoagulants (NOAC) have heterogeneity. Available evidence reveals that any heart failure and/or left ventricular systolic dysfunction is associated with higher rates of stroke/systemic embolism and bleeding in patients with non valvular atrial fibrillation compared to patients without heart failure and normal left ventricular function. Most standard dose NOAC regimens have a better safety and efficacy profile over warfarin in most heart failure sub-group types with a few exceptions including patients with NYHA III/IV on Dabigatran 150mg BID from the RE-LY trial, who had higher major bleeding events, and patients with asymptomatic left ventricular dysfunction (ejection fraction ≤40%) and heart failure with reduced ejection fraction on 20mg of Rivaroxaban in the ROCKET-AF trial, when compared to patients on Warfarin in the corresponding groups. With the gaining popularity and use of NOACs, understanding their safety profile in such situations is paramount.

Effect of peripheral administration of cinnarizine and verapamil on the abstinence syndrome in diazepam-dependent rats.

The effects of two calcium channel blockers (verapamil and cinnarizine) were evaluated on diazepam withdrawal symptoms. Rats were made diazepam dependent by chronic treatment with daily injections of the drug, 20 mg/kg IP for 3 weeks. On abrupt termination of the drug, animals showed withdrawal hyperactivity that was assessed by autonomic, behavioural and motor signs. The peak effect was seen 3 days after the withdrawal of diazepam. On IP administration, verapamil and cinnarizine (10, 20 and 40 mg/kg) given on eight occasions at an interval of 12 h reversed the withdrawal-induced increase in spontaneous motor activity. Cinnarizine in higher doses (20 and 40 mg/kg) was found to be effective in suppressing the behavioural signs but verapamil did not show any protective effect against startle response and irritability. These results suggest that modulation of the calcium influx in the CNS might influence withdrawal.

Diazepam-atenolol combination antagonizes aminophylline-induced convulsions and lethality in mice.

The present study was undertaken to identify protective drugs against aminophylline (240 mg/kg i.p.)-induced convulsions and lethality in mice. Diazepam (10 mg/kg) and valproic acid significantly prevented the convulsions, but were not effective in preventing mortality. Phenytoin, atropine, carbamazepine and atenolol were ineffective in protecting against convulsions and death. Ketamine gave partial protection against convulsions, but was not effective in preventing mortality. Diazepam (10 mg/kg) and atenolol (5 mg/kg) administered together gave total protection against convulsions and death. These results show that aminophylline-induced convulsions are relatively resistant to antiepileptic drugs, and that a combination of diazepam and a beta-blocker (atenolol) has potential as an anti-aminophylline agent.

Search for an antidote to electroconvulsion and lethality in aminophylline-sensitized mice.

Diazepam (10 mg/kg i.p.) or MK-801 (0.25 mg/kg i.p.) offered complete protection against corneal electroshock (30 mA x 0.2 s)-induced tonic seizures and lethality but failed to protect from aminophylline (150 mg/kg i.p.) + electroshock (15 mA x 0.2 s)-induced tonic seizures and lethality in mice. The diazepam (2.5 mg/kg i.p.) and MK-801 (0.25 mg/kg i.p.) combination completely protected the mice from aminophylline + electroshock-induced seizures and lethality. Sodium valproate (500 mg/kg i.p.) protected the mice from electroshock (30 mA) per se and aminophylline + electroshock (15 mA)-induced seizure and lethality. The present study established the neurosensitizing potential of a single, non-convulsive dose of aminophylline for electroconvulsion due to subthreshold intensity electroshock and demonstrated the prophylactic efficacy of sodium valproate and the synergistic therapeutic potential of diazepam and MK-801 combination against such seizure attacks.

Memory enhancing effects of granisetron (BRL 43694) in a passive avoidance task.

Recent evidence suggests that serotonin plays an important role in learning and memory processes in animals. The present study examined the effect of the 5-HT3 receptor antagonist, granisetron (BRL 43694), on acquisition, retention and retrieval of a passive avoidance response in mice. Granisetron (1 and 10 micrograms/kg) administered 30 min before presentation of footshock increased the step-down latency when tested 24 h after footshock. The acquisition process was not affected by a dose of 100 micrograms/kg. Granisetron (10 and 100 micrograms/kg) produced a significant increase in latency to step out of the safety zone, when administered immediately after or 23.5 h after footshock. However, at 1 microgram/kg, granisetron had no effect. These results confirm the important role played by 5-HT in the process of learning and memory, and also suggest that memory enhancement may be possible with non-cholinergic treatments.

MK-801 antagonizes the lethal action of centrally and peripherally administered cypermethrin in mice and rats.

The present study investigated the effect of MK-801, an N-methyl-D-aspartate antagonist, on the convulsant lethal action of cypermethrin administered centrally or peripherally. Cypermethrin produced severe convulsions and death in a dose-dependent manner. MK-801 (0.5, 1 and 2 mg kg-1, intraperitoneally) significantly increased the onset time of convulsions and decreased the mortality in the peripherally treated cypermethrin group. MK-801 (1.0 and 2.0 mg kg-1) attenuated the convulsant action of cypermethrin (50 micrograms, intracerebroventricularly) significantly. Survival rate was also increased significantly. However, MK-801 (0.5 mg kg-1) did not produce any significant protective effect against centrally administered cypermethrin. These results suggest excitatory amino acids to be a target for pyrethroid-induced neurotoxicity.

Interactions of verapamil and diltiazem with ketamine: effects on memory and sleeping time in mice.

The effects of ketamine (3, 10 and 30 mg/kg) alone and in combination with verapamil (10 mg/kg) or diltiazem (30 mg/kg) on the acquisition, consolidation and retrieval of memory using a passive avoidance task in mice were studied. Ketamine significantly inhibited the acquisition and consolidation of memory at 10 and 30 mg/kg dose levels and these effects were antagonized by diltiazem 30 mg/kg but not by verapamil 10 mg/kg. Studies of sleeping time demonstrated that pretreatment with verapamil 10 mg/kg increased the duration of sleeping time. Diltiazem, however, did not potentiate the effects of ketamine on sleeping time. The present findings indicate that diltiazem can counter the effects of ketamine on memory. The data also indicates that pretreatment of surgical patients with verapamil may reduce the dose of ketamine required for anesthesia.

Possible mechanism of haloperidol-induced enhancement of memory retrieval.

The effects of pretest administration of haloperidol on retention of a step-through passive avoidance task was studied with a 3-day training and retention test interval. Haloperidol at doses of 0.3 mg/kg and 0.5 mg/kg i.p. enhanced memory retrieval. This effect of haloperidol is possibly mediated by interaction with alpha 2-adrenergic receptors, that results in increasing the noradrenergic transmission, or it could be due to direct interaction with dopamine receptors. Yohimbine (2.5 mg/kg) also enhanced memory retrieval, as shown by haloperidol. Clonidine (0.05 mg/kg) significantly antagonized the effect of haloperidol on memory retrieval. Scopolamine (0.5 mg/kg) did not have any effect on enhancement of memory produced by haloperidol.

Effect of subacute DDT on pharmacokinetics of isoniazid and liver function in rabbits.

DDT administration (30 mg/kg per day, po, for 21 consecutive days) to rabbits showed an increase in peak plasma concentration and a decrease in time to reach peak plasma concentration of isoniazid whereas no change was observed in elimination rate constant and area under the plasma concentration-time curve. DDT treatment caused increased absorption of isoniazid. Early signs of hepatic damage were also observed. Since there was no change in the levels of serum glutamate oxaloacetate transaminase and serum glutamate pyruvate transaminase, it can be concluded that DDT does not significantly affect liver function at the dosage used. The observed elevated levels of alkaline phosphatase could be due to direct activation of the enzyme. Leukopaenia and neutropaenia with relative lymphocytosis indicated that DDT might have suppressant effect on granulocyte cell line of WBCs.

Possible mechanism of endosulfan-induced enhancement of memory acquisition and retention in mice.

The effects of pre-training and post-training administration of endosulfan on retention of a step-down passive avoidance task was studied in mice. Endosulfan at doses of 1.0 mg/kg(ip) and 2.0 mg/kg(ip) enhanced memory acquisition and retention. This effect of endosulfan was possibly mediated by interaction with cholinergic neurotransmission, as scopolamine (0.5 mg/kg, ip) significantly antagonized the memory enhancing effects of endosulfan. Clonidine (0.05 mg/kg, ip) did not have any effect on enhancement of memory produced by endosulfan, thus indicating possibly no role of noradrenergic system.

Effects of post-training administration of (-)-baclofen and chlordiazepoxide on memory retention in ICRC Swiss mice: interactions with GABAA and GABAB receptor antagonists.

The effects of post-training administration of chlordiazepoxide and (-)-baclofen on memory retention was studied in ICRC Swiss mice by measuring the retest stepdown latency 24 hr after foot-shock in a passive avoidance task. Chlordiazepoxide 20 mg/kg impaired memory retention and a similar effect was produced by 10 mg/kg of diazepam. The effect of chlordiazepoxide was antagonised when combined with picrotoxin but not by the addition of a specific GABAB antagonist CGP 35348. The effect of chlordiazepoxide on memory retention seems to be mediated by action at the GABAA-benzodiazepine receptor complex. (-) Baclofen, the active isomer of the GABAB agonist enhanced memory in ICRC mice and this effect was antagonised by CGP 35348 at a dose of 10 mg/kg. The inactive isomer of baclofen, (+)-baclofen did not produce any effect. This indicates that GABAB receptors contribute to the effects of (-)-baclofen on memory.

Enhancement of memory retrieval and attenuation of scopolamine-induced amnesia following administration of 5-HT3 antagonist ICS 205-930.

Experimental evidence suggests an important role of serotonin in the process of learning and memory. The present study investigated the effect of 5HT3-receptor antagonist (ICS 205-930) on retrieval of a previously learned aversive habit in the mouse. The effect of ICS 205-930 on scopolamine (3 mg/kg) induced amnesia was also studied. ICS 205-930 (1, 10 & 100 micrograms/kg) produced a dose-dependent increase in latency to cross into the dark chamber. The scopolamine induced memory impairment was significantly attenuated by ICS 205-930 (10 micrograms/kg). These results suggest that memory deficits may be susceptible to attenuation with non-cholinergic treatments.

BRL 38227--a potassium channel opener, antagonizes digoxin-induced convulsions.

Experimental evidence suggests that potassium channel openers play an important role in convulsions. In this study, the anticonvulsant activity of BRL 38227, a new potassium channel opener against digoxin-induced convulsions, is reported. Intraventricular administration of digoxin (7.5 micrograms), included "popcorn-type" convulsions in rats. BRL 38227, injected centrally increased the onset time of convulsions and decreased the mortality rate in a dose-dependent manner. Pretreatment with 4-aminopyridine, a potassium channel blocker antagonized the protective effect of BRL 38227. These findings show the involvement of potassium channels in digoxin-induced convulsions. Further these results indicate that in the future potassium channels might be a target for new anticonvulsant drugs.

Comparative bioavailability of two enteric-coated capsules of omeprazole in healthy volunteers.

A comparative bioavailability study was carried out on two enteric-coated capsules (20 mg each) of omeprazole (omeprazole, Alembic: "A" and Losec, Astra, England: "B"). The in-vitro dissolution of both products "A" and "B" met the prescribed USP standard. The bioavailability of single dose (20 mg) and multiple doses (20 mg once daily for 7 days) of both products "A" and "B" were carried out in eight healthy male volunteers in a crossover design. The rate and extent of bioavailability of omeprazole was higher in product "A" following a single oral dose, suggesting its therapeutic advantage over the product "B" in the prevention of acid aspiration during surgery. In multiple dose study, the two products were found bioequivalent as assessed by AUC0-infinity, Cmax, tmax and t1/2 elimination.