Safety and probiotic evaluation of two Bacillus strains producing antioxidant compounds.

Bacillus species are becoming increasingly relevant for use as probiotics or feed additives where their heat stability can ensure survival in the food matrix or enable long-term storage at ambient temperature. Some Bacillus species are pigmented and in this study, we have examined two strains, one Bacillus pumilus (pigmented red) and the other Bacillus megaterium (pigmented yellow) for their safety for potential use in humans as dietary supplements. In addition, we have set out to determine if they might confer any potential health benefits. Both strains produce C30 carotenoids while the B. pumilus strain also produced large quantities of riboflavin equivalent to genetically modified Bacillus strains and most probably contributing to this strain's pigmentation. Riboflavin's and carotenoids are antioxidants, and we have evaluated the ability of vegetative cells and/or spores to influence populations of Faecalibacterium prausnitzii in the colon of mice. While both strains increased levels of F. prausnitzii, spores of the B. pumilus strain produced a significant increase in F. prausnitzii levels. If found to be reproducible in humans such an effect might, potentially, confer health benefits particularly for those suffering from inflammatory bowel disease.

Study of 7 Cases of Giant Cell Tumor of Soft Tissue.

BACKGROUND: Primary giant cell tumour of soft tissues is a distinct but uncommon group of neoplasms morphologically identical to osseous giant cell tumor. METHODS: 7 patients with painless growing soft tissue mass, having no attachment to underlying bone, were identified in a four years retrospective study from two zonal hospitals of armed forces. Histopathology of these lesions revealed admixture of multinucleated giant cell with mononuclear cells. All patients were treated by surgical resection and followed up for recurrence. Results : There were 5 male and 2 female patients in the age group of 18 to 56 years. All lesions were superficial, circumscribed and involved extremities except one. Histologic transition between benign and malignant lesion was present in only one of the 7 patients that recurred after three months of surgery for which she had to be operated again. 2 of our 7 cases were lost in follow up. CONCLUSION: Primary giant cell tumour of soft tissues usually present as a painless mass and needs to be differentiated from other giant cell rich soft tissue tumors. Benign clinical course is expected if the lesion is excised adequately. Its biological behaviour to have low malignant potential is recognized; but this cannot be predicted and metastasis does occur rarely.

Expression of PPAR, RXR isoforms and fatty acid transporting proteins in the rat and human gastrointestinal tracts.

Dietary fatty acid (FA) absorption across the gastrointestinal (GI) tract is of critical importance for sustenance, however, excessive FA absorption has also been linked to metabolic syndrome and associated disorders. The expression of isoforms that regulate the dietary FA absorption are not as well characterized in the GI tract as they are elsewhere. Peroxisome proliferator-activated receptors (PPARalpha, beta, and gamma) and 9-cis-retinoic acid receptors (RXRalpha, beta, and gamma) are nuclear hormone transcription factors that control FA homeostasis, in part through the regulation of expression of membrane-bound FA transporting proteins. The present study was designed to elucidate the expression of PPAR and RXR isoforms and FA transporting proteins (FABPpm and FAT/CD36) in the rat and human GI tracts using reverse transcriptase-polymerase chain reaction (RT-PCR), immunoblotting, and immunohistochemical staining. The results revealed rat GI expression of all the PPAR and RXR isoforms, FABPpm and FAT/CD36. PPARalpha, PPARbeta, PPARgamma, RXRalpha, FABPpm, and FAT/CD36 isoforms exhibited ubiquitous expression in human GI tract, whereas RXRbeta was not detected. RXRgamma was observed in a majority of the human GI samples. These results provide a physiological foundation for rational drug design and drug delivery for the mitigation of metabolic syndrome and associated disorders to normalize intestinal FA absorption.

Comparison of analgesic and anti-inflammatory activity of meloxicam gel with diclofenac and piroxicam gels in animal models: pharmacokinetic parameters after topical application.

Meloxicam, a non-steroidal anti-inflammatory drug, is a preferential inhibitor of cyclooxygenase-2 and has demonstrated potent analgesic and anti-inflammatory activity after oral administration. The present work was carried out to elucidate the anti-inflammatory and analgesic activity of a newer topical gel formulation of meloxicam (1% w/w gel) and compare it with 0.5% w/w piroxicam and 1% w/w diclofenac gels in experimental animal models. The study was also extended to determine the pharmacokinetic profile of a newer formulation of meloxicam gel after topical application on depilated skin of rats. The anti-inflammatory activities of meloxicam, piroxicam and diclofenac gels were compared using carrageenan-induced acute paw oedema and complete Freund's adjuvant-induced chronic paw oedema in rats. Meloxicam gel showed increased protection against inflammation as compared to piroxicam and diclofenac gels. Acetic acid-induced writhing and formalin-induced phase I and phase II pain models were used to compare their analgesic activity. Meloxicam gel showed significant protection in formalin-induced phase II pain whereas its analgesic activity was less as compared to diclofenac and piroxicam gels in writhing test and formalin-induced phase I pain. The pharmacokinetic studies showed peak plasma drug concentration (C(max)) of 48.48 +/- 6.57 microg/ml at 2 h (T(max)) after topical application of 500 mg of meloxicam gel formulation. The area under the curve as calculated from 0 to 6 h was found to be 114.18 +/- 4.23 and 194.13 +/- 3.78 microg x h/ml for 0 to infinity. The results indicate that topical preparation of meloxicam could be an effective alternative to diclofenac and piroxicam gels in inflammatory conditions and its associated pain with the possibility of less systemic side-effects.

Piperine in food: interference in the pharmacokinetics of phenytoin.

This study was carried out to explore the effect of piperine-containing food in altering the pharmacokinetics of phenytoin, an anti-epileptic drug with a narrow therapeutic index. A preliminary pharmacokinetic study was carried out in mice by administering phenytoin (10 mg) orally, with or without piperine (0.6 mg). Subsequently, oral pharmacokinetics of phenytoin was carried out in six healthy volunteers in a crossover design. Phenytoin tablet (300 mg) was given 30 minutes after ingestion of a soup (melahu rasam) with or without black pepper. A further study of intavenous pharmacokinetics of phenytoin (1 mg) in rats with or without oral pretreatment with piperine (10 mg) was also conducted. The phenytoin concentration in the serum was analyzed by HPLC. The study showed a significant increase in the kinetic estimates of Ka, AUC(0-10) and AUC(0-infinity) in the piperine-fed mice. Similarly, in human volunteers piperine increased Ka, AUC(0-48), AUC(0-infinity), and delayed elimination of phenytoin. Intravenous phenytoin in the oral piperine-treated rat group showed a significant alteration in the elimination phase indicating its metabolic blockade. The significance of this finding in epileptic patients maintained on phenytoin therapy requires further investigation. This study may also have implications in the case of other drugs having a low therapeutic index.

Comparative anti-nociceptive, anti-inflammatory and toxicity profile of nimesulide vs nimesulide and piperine combination.

Piperine is an inhibitor of various hepatic and other enzymes involved in the biotransformation of drugs. Preliminary pharmacokinetic studies conducted by us suggested the increased bioavailability of nimesulide co-administered with piperine. The present study was, thus, conducted to evaluate the antinociceptive, anti-inflammatory and toxicity profile of a new nimesulide-piperine combination administered orally as compared with nimesulide alone. Antinociceptive efficacy was tested using an acetic acid writhing test and tail flick latency test (TFL). The ED50 value of a nimesulide-piperine combination in writhing test was calculated to be significantly lower (1.5 mg kg(-1)) as compared to (11.2 mg kg(-1)) of nimesulide alone. The antinociceptive effect was lesser in the tail flick latency test as compared to what was observed in the writhing test indicating the peripheral action of the Non-Steriodal Anti-Inflammatory Drug (NSAID). In carrageenan-induced inflammatory tests, the nimesulide-piperine combination was found to be dose-to-dose superior than nimesulide alone. Acute toxicity studies on mice revealed a reduction in lethal dose (LD50) of the combination (980 mg kg(-1)) as compared to nimesulide (1500 mg kg(-1)) alone. Results from the present study suggest a better therapeutic index for the nimesulide-piperine combination indicating that this combination would further reduce the frequency of adverse effects associated with nimesulide alone.

Development and validation of a new high-performance liquid chromatographic estimation method of meloxicam in biological samples.

A simple, HPLC method was developed to estimate meloxicam (COX-2 inhibitor) using piroxicam as the internal standard. The mobile phase containing methanol, acetonitrile and an aqueous solution of diammonium hydrogenorthophosphate (50 mM) in the ratio of 4:1:5 was pumped at the rate 1 ml/min. Lichrocart RP-18 (125 x 4 mm) was used as an analytical column and the analytes were detected at 364 nm using a UV detector. Acidified plasma samples were extracted with chloroform, evaporated to dryness, reconstituted in the mobile phase and then a volume of 10 microl of the prepared sample was injected in the column. The retention time of meloxicam and piroxicam was found to be 2.7 and 1.9, respectively. This method showed an accuracy of 102.3% at 0.52 microg/ml and was capable of detecting a minimum concentration of 0.029 microg/ml meloxicam from biological samples. The analytical method was successfully utilized for estimating meloxicam in biological samples.

Anti-inflammatory activity and pharmacokinetic profile of a new parenteral formulation of nimesulide.

Nimesulide, a selective COX-2 inhibitor, exerts potent anti-inflammatory and analgesic effects when administered orally, rectally or topically. The present study was designed to evaluate the anti-inflammatory activity of a new parenteral formulation of nimesulide and to correlate it with the pharmacokinetic profile. Nimesulide was administered intramuscularly at increasing doses of 1. 5, 3, 6, 12.5 and 25 mg kg-1 which produced dose-dependent anti-inflammatory effects in the carrageenan-induced rat paw edema. The anti-inflammatory activity of nimesulide was greater than that of diclofenac which was administered at identical doses though the difference was not statistically significant. Peak anti-inflammatory effects with nimesulide were observed between 2 and 3 h post-treatment which correlates well with the tmax of 115 min. The plasma concentration of nimesulide at different time points was assayed using HPLC after administration at a dose of 25 mg kg-1. Peak plasma concentration (Cmax) was 23 microgram ml-1 while t1/2 was derived as 4.2 h. Area Under Curve (AUC(0-6 h)) was calculated as 83. 31 microgram ml-1 h-1. No toxicity or adverse effects were noted at the doses administered. The present study demonstrates that nimesulide administered intramuscularly may be superior to other routes of administration when fast onset of action is required with high plasma concentration.