Assessment of antibacterial activity of smaller chain tripeptides and tetrapeptides.

Our objective is to investigate the smaller chain tripeptides and tetrapeptides with the efforts mainly directed towards the identification of compounds presenting a high bactericidal activity. In this connection, 5 peptides, Met-Arg-Tyr (MRY), Met-Val-Tyr (MVY), Met-Ile-Cys-Tyr (MICY), Phe-Trp-Lys-Tyr (FWKY) and Met-Trp-Lys-Tyr (MWKY) were synthesized by solid phase peptide synthesis. The column eluted pure synthesized compounds were tested for in-vitro antibacterial disc diffusion assay using 2 g positive S. aureus, B.subtilis and 1 g negative E.coli bacterial strains at different concentrations predicted by pH and inhibitory concentration findings. 2 of the tetrapeptides, Phe-Trp-Lys-Tyr (FWKY) and Met-Trp-Lys-Tyr (MWKY) were found to be highly efficient against all the gram positive and microbial strains tested. Maximum activity was observed at a concentration of 300 µg/ml (499.17 µM) for the tetrapeptide Phe-Trp-Lys-Tyr (FWKY) against B. subtilis and at a concentration of 450 µg/ml (748.75 µM) against E. coli with the corresponding zonal inhibition diameter readings (17 mm; 16 mm). The tetrapeptide Met-Trp-Lys-Tyr (MWKY) was found to have high potency against S. aureus at a concentration of 450 µg/ml (769.23 µM) with the corresponding zonal inhibition diameter as 13 mm. The experimental results are analysed statistically by t-test at 5% and 1% level of significance. Our tetrapeptides have shown antibacterial action against both gram positive and gram negative strains and are considered safer as par with commercial antibiotics available today. Further clinical studies will make sure to position our potent small chain tetrapeptides in the arsenal of new broad spectrum anti-gram positive and anti-gram negative agent.

Antimicrobial glycopeptides: synthesis and antibacterial activity of N-linked and O-linked smaller chain glycopeptides.

Our objective is to synthesize the smaller chain N-linked and O-linked glycopeptides using sugars belong to mono, di and polysaccharides, with the efforts mainly directed towards the identification of antibacterial compounds. 7 glycopeptides, viz., Arg-Asn-Mannose, Arg-Asn-Lactose, His-Asn-Mannose, His-Asn-Lactose (N-glycopeptides), Arg-Ser-Lactose, Arg-Thr-Lactose, Arg-Thr-Starch (O-glycopeptides) were prepared by dicyclohexyl carbodimide (DCC) coupling for amino acids using microwave oven (50W power; 15 min) and activated and coupled with respective sugar moieties using microwave oven at 120 W for 20-25 min. The column eluted compounds were tested for disc diffusion assay using 3 gram positive S. aureus, B. subtilis, S. caprae and 3 gram negative E. coli, P. aeruginosa and S. sonnei strains at different concentrations predicted by pH and inhibitory concentrations. One of the test glycopeptide, His-Asn-Lactose was found to be very effective against all the microbial strains tested and 3 other Test Compounds, viz., His-Asn-Mannose, Arg-Thr-Lactose and Arg-Thr-Starch are also proved to be effective against 2 gram positive and 2 gram negative strains tested. Maximum activity was observed at a concentration of 450 µg/ml (747.51 µM) for the N-glycopeptide His-Asn-Lactose with the corresponding zonal inhibition diameters (15 mm; 19 mm; 14 mm; 18 mm; 16 mm; 17 mm) against S. aureus, B. subtilis, S. caprae, E. coli, P. aeruginosa, S. sonnei. This is the first evidence based report that our N-glycopeptide, His-Asn-Lactose tested, has shown antibacterial action against both gram positive and gram negative strains. Further in vivo testing and clinical studies will make sure to position our potent small chain N-glycopeptide in the arsenal of new broad spectrum anti-gram positive and negative agent.

Dicer is required for proliferation, viability, migration and differentiation in corticoneurogenesis.

In mice, microRNAs (miRNAs) are required for embryonic viability, and previous reports implicate miRNA participation in brain cortical neurogenesis. Here, we provide a more comprehensive analysis of miRNA involvement in cortical brain development. To accomplish this we used mice in which Dicer, the RNase III enzyme necessary for canonical miRNA biogenesis, is depleted from Nestin-expressing progenitors and progeny cells. We systematically assessed how Dicer depletion impacts proliferation, cell death, migration and differentiation in the developing brain. Using markers for proliferation and in vivo labeling with thymidine analogs, we found reduced numbers of proliferating cells, and altered cell cycle kinetics from embryonic day 15.5 (E15.5). Progenitor cells were distributed aberrantly throughout the cortex rather than restricted to the ventricular and subventricular zones. Activated Caspase3 was elevated, reflecting increased cortical cell death as early as E15.5. Cajal-Retzius-positive cells were more numerous at E15.5 and were dysmorphic relative to control cortices. Consistent with this, Reelin levels were enhanced. Doublecortin and Rnd2 were also increased and showed altered distribution, supporting a strong regulatory role for miRNAs in both early and late neuronal migration. In addition, GFAP staining at E15.5 was more intense and disorganized throughout the cortex with Dicer depletion. These results significantly extend earlier works, and emphasize the impact of miRNAs on neural progenitor cell proliferation, apoptosis, migration, and differentiation in the developing mammalian brain.

Development and physical characterization of chloramphenicol loaded biodegradable nanoparticles for prolonged release.

The objectives of our study were to prepare a biodegradable nanoparticulate system of chloramphenicol (CHL) and to evaluate its ability to prolong in vitro release of CHL compared to free drug suspension (FDS). CHL-loaded polylactide-co-glycolide nanoparticles (CHL-PLGA-NPs) were prepared by an emulsion/solvent evaporation method using ethyl acetate and polyvinyl alcohol. CHL-PLGA-NPs were characterized by particle size, zeta potential, infrared spectra, drug entrapment efficiency and in vitro release kinetics measurement. Sonication was done with an ultrasound pulse sonicator at 70 W, 30 kHz for 60 s to produce stable NPs of mean size range from 277 nm to 433 nm. Drug to polymer ratio (D:P) was selected as formulation variable and significantly influenced entrapment efficiency (approximately 30% to 66%) and release (p < 0.05). Entrapment of CHL in biodegradable NPs significantly prolonged drug release compared to FDS and thus implies potential antibiotic delivery system for ocular application.

Metal concentrations in water, sediment, and fish from sewage-fed aquaculture ponds of Kolkata, India.

The concentrations of lead, cadmium, chromium, copper, and zinc were investigated in the sewage-fed pond water, sediment, and the various organs of Labeo rohita, Catla catla, Cirrhinus mrigala, Oreochromis mossambicus, and Cyprinus carpio cultured in sewage-fed ponds, Kolkata, India. Among the metals, cadmium, lead, and zinc were detected in water and, except lead, were below the water quality guideline levels for the protection of freshwater aquatic life proposed by CEQG (Canadian Environmental Quality Guidelines) and AENV (Alberta Environment). Therefore, lead could pose danger to aquatic organisms. All the five metals were detected in the sediment and, except cadmium and lead, were below the sediment quality guideline levels for aquatic life proposed by EPA (Environmental Protection Agency). Therefore, these two metals could be toxic to aquatic organisms. Significant (P > 0.05) differences were observed among the five fish species for all these metals accumulation. Also, significant (P > 0.05) differences were noticed among these metals accumulation in fish organs. Cadmium showed the least bioaccumulation, while zinc showed the highest bioaccumulation in all the fish species. Though the metal concentration in the different fish tissues was variable, the highest concentration was found in kidney and the lowest in the muscle. Concentrations of these metals in the muscle tissue of all the fish species were well below the consumption safety tolerance in fish set by WHO/FAO, and thus, so far as these metals are concerned, these sewage-fed cultured fishes are safe and suitable for human consumption.

Antidiabetic Activity of the Ethanol Extract of Capparis sepiaria L Leaves.

Capparis sepiaria L, a profusely branched hedge plant, is used in Indian traditional medicine. Capparis sepiaria leaves were extracted with ethanol and concentrated to dryness. The LD(50) value was determined as 894.43 mg/kg body weight by acute toxicity study. The ethanol extract was investigated for possible hypoglycemic effect produced by single oral administration at various dose levels 100, 200 and 300 mg/kg in the streptozotocin induced diabetic rats and compared against normal saline control and the standard glibenclamide. A maximum fall of plasma glucose level 9.40%; 13.57%; 15.25% and 18.80% was observed after 12 h of treatment when administered with ethanol extract of Capparis sepiaria at 100, 200 and 300 mg/kg, and glibenclamide 10 mg/kg dose, respectively. The findings from the study suggest that the Capparis sepiaria leaves may be prescribed as an adjunct to traditional formulation and drug treatment for controlling diabetes mellitus.

A validated UV spectrophotometric method for determination of duloxetine hydrochloride.

A simple, sensitive and accurate UV spectrophotometric method was developed for the assay of duloxetine hydrochloride in raw material and capsules. Validation of the method yielded good results concerning range, linearity, precision and accuracy. The absorbance was measured at 290 nm for duloxetine capsule solution. The linearity range was found to be 5-50 microg/mL for the drug. It was found that the excipients in the commercial formulation did not interfere with the methods.

A validated UV spectrophotometric method for estimation of nebivolol hydrochloride in bulk and pharmaceutical formulation.

A simple, sensitive and accurate UV spectrophotometric method was developed for the assay of nebivolol hydrochloride in raw material and tablets. Validation of the method yielded good results concerning range, linearity, precision and accuracy. The absorbance was measured at 282 nm for nebivolol hydrochloride tablet solution. The linearity range was found to be 5-50 microg/mL for the drug. It was found that the excipients present in the commercial formulation did not interfere with the method.

Comparative anti - microbial evaluation studies of the extracts and isolates of leaves & bark of wrightia tomentosa.

The Butanol and Ethanol extract of the leaves and bark of Wrightia tomentosa along with its seven pure component isolates (BLF(28), BLF(29*), BBF(29), ELF(3), ELF7, ELF(17*), EBF(7) ) after fractionation by column chromatography were evaluated for antimicrobial activity against Gram positive (S. aureus, S. fecalis, S.albus and B.subtilis) and Gram negative (Escherichia coli, Pseudomonas aeruginosa, Proteus vulgaris & Klebsiella aerogenes) bacteria and the fungi Candida albicans by disc diffusion method. The extracts and isolates showed different degree of activity against pathogenic microbes. The results obtained were compared with standard drugs Ciprofloxacin (10µg) and Clotrimazole (10µg). The isolates of butanol bark extract (BBF(29)) followed by leaf extract(BLF(29*)) were considerably more effective than the ethanol leaf and bark extract in inhibiting all the microbial strains.

Pharmacognostical studies on leaves of commiphora caudata (wight & arn) engl.

Commiphora caudata (Wight & Arn) is a potential medicinal plant used for its antispasmodic activity, cytotoxic activity and hypothermic activity. Owing to its medicinal importance, macroscopic and microscopic characters of leaves of Commiphora caudata were studied.

Structural changes of beta-lactoglobulin during thermal unfolding and refolding--an FT-IR and circular dichroism study.

We have quantitatively characterized by FT-IR spectroscopy the contents of secondary structure of beta-lactoglobulin during thermal unfolding and subsequent refolding. Our data clearly indicate that considerable amount of secondary structure, particularly beta-sheet, still remained intact even at 90 degrees C. Noticeable changes in secondary structure of beta-lactoglobulin were observed only above 70 degrees C. The refolded protein regained, within limits of experimental error, all of the secondary structure lost during thermal unfolding. The data also indicate that the refolding mechanism operating at pH 7.0 and 2.0 are the same. Identical secondary structure of native and refolded beta-lactoglobulin was also indicated by far-UV circular dichroic spectra of the two forms of protein. Near UV circular dichroic spectra of the same two forms showed considerable differences indicating less tertiary structure of refolded beta-lactoglobulin. The combined CD and FT-IR data indicated that refolded form of beta-lactoglobulin could be characterized as a molten globule state as it had native-like secondary structure and compromised tertiary structure.

Anomalous magnetic properties of mechanically milled cobalt oxide nanoparticles.

Defect induced magnetic properties of CoO nanoparticles produced via mechanical ball milling have been assessed by detailed magnetic measurements. A progressive decrease in the particle size and a concomitant increase in the induced strain have been observed with the milling times. The mechanically milled nanoparticles of CoO exhibit anomalous magnetic properties such as FM hysteresis when compared with the unmilled CoO sample. The presence of weak ferromagnetism, with a highest value of magnetization of 0.532 emu/g at 10 K in the 100 h milled sample, is attributed to the uncompensated surface spins resulting from induced surface defects via mechanical milling. The ZFC coercive force, measured at 10 K, increases with milling time reaching a maximum value of 1066 Oe for the 100 h milled sample. The temperature dependent field-cooled (FC) and zero-field-cooled (ZFC) magnetic measurements indicate a presence of an exchange bias field arising from uncompensated moments generated by mechanical strain and the antiferromagnetic (AFM) core. The exchange bias field measured at 10 K reaches a value 210 Oe for the 50 h milled sample and decreases upon prolonged milling. The exchange bias field vanishes at a temperature approximately 200 K, a temperature much lower than the Neel temperature of CoO (TN approximately 291 K). The observed anomalous magnetic behavior of CoO could be interpreted in terms of the exchanged bias FM-AFM model.

Evaluation of purgative activity of roots of Rumex nepalensis.

The methanol extract of Rumex nepalensis Spreng. roots at the oral dose of 100-400 mg/kg exhibited significant and dose-dependent purgative activity.

Topographical distribution of laryngeal carcinoma - A study of fifty cases.

Regional and site-specific distribution of fifty cases of Laryngeal carcinoma is presented along with a review of the avialable literature.

Oxidative DNA damage in prostate cancer patients consuming tomato sauce-based entrees as a whole-food intervention.

BACKGROUND: Human prostate tissues are vulnerable to oxidative DNA damage. The risk of prostate cancer is lower in men reporting higher consumption of tomato products, which contain high levels of the antioxidant lycopene. We examined the effects of consumption of tomato sauce-based pasta dishes on lycopene uptake, oxidative DNA damage, and prostate-specific antigen (PSA) levels in patients already diagnosed with prostate cancer. METHODS: Thirty-two patients with localized prostate adenocarcinoma consumed tomato sauce-based pasta dishes for the 3 weeks (30 mg of lycopene per day) preceding their scheduled radical prostatectomy. Serum and prostate lycopene concentrations, serum PSA levels, and leukocyte DNA oxidative damage (ratio of 8-hydroxy-2'-deoxyguanosine [8-OHdG] to 2'-deoxyguanosine [dG]) were assessed before and after the dietary intervention. DNA oxidative damage was assessed in resected prostate tissue from study participants and from seven randomly selected prostate cancer patients. All statistical tests were two-sided. RESULTS: After the dietary intervention, serum and prostate lycopene concentrations were statistically significantly increased, from 638 nM (95% confidence interval [CI] = 512 to 764 nM) to 1258 nM (95% CI = 1061 to 1455 nM) (P<.001) and from 0.28 nmol/g (95% CI = 0.18 to 0.37 nmol/g) to 0.82 nmol/g (95% CI = 0.57 to 1.11 nmol/g) (P <.001), respectively. Compared with preintervention levels, leukocyte oxidative DNA damage was statistically significantly reduced after the intervention, from 0.61 8-OHdG/10(5) dG (95% CI = 0.45 to 0.77 8-OHdG/10(5) dG) to 0.48 8-OHdG/ 10(5) dG (95% CI = 0.41 to 0.56 8-OHdG/10(5) dG) (P =.005). Furthermore, prostate tissue oxidative DNA damage was also statistically significantly lower in men who had the intervention (0.76 8-OHdG/10(5) dG [95% CI = 0.55 to 0.96 8-OHdG/10(5) dG]) than in the randomly selected patients (1.06 8-OHdG/10(5) dG [95% CI = 0.62 to 1.51 8-OHdG/10(5) dG]; P =.03). Serum PSA levels decreased after the intervention, from 10.9 ng/mL (95% CI = 8.7 to 13.2 ng/mL) to 8.7 ng/mL (95% CI = 6.8 to 10.6 ng/mL) (P<.001). CONCLUSION: These data indicate a possible role for a tomato sauce constituent, possibly lycopene, in the treatment of prostate cancer and warrant further testing with a larger sample of patients, including a control group.

Product development studies for stabilization of oral liquid enzyme preparations containing diastase and papain.

A product development study was undertaken with a view to stabilize acid sensitive amylolytic enzyme diastase and alkali sensitive proteolytic enzyme papain. An unique high technology method was adopted for both enzymes with enteric coating and non-enteric film forming materials respectively to achieve site specificity of action, better stability and to protect diastase from the adverse acid pH of the stomach. Feasibility of incorporating various excipients like thickening agents, surfactants, preservatives, etc., to produce stable elegant oral liquid suspension was also studied. Release studies of the enzymes both in simulated gastric and simulated intestinal juices showed steady and consistent release. The proteolytic and amylolytic activities were assessed by standard pharmacopoeial methods. The stability of the product during the studies was satisfactory. Reduction in addition of overages of costly enzymes met the project technically feasible and economically viable.

Computer supported studies on design and evaluation of solid dispersions of carbamazepine.

The main advantages of solid dispersions (the drug could be maintained in a bioavailable form, dosage reduction and cleaner manufacturing conditions) provide scope for the continued interest in field. Additionally, their use in providing a sustained or controlled release of drugs has only been tentatively examined. In the present study, Carbamazepine has been used to develop a dosage form which will provide a booster amount followed by sustained release of the drug for effective control of epileptic seizures while keeping the serum level of the drug at minimum. Enteric polymers CAP and CAT have been used to retard the release till the formulation reaches the intestine. Either individually or combined fractions of the formulations may be used in the therapy of epilepsy.

Preparation and in vitro dissolution of isoniazid from ethylcellulose microcapsules.

Microcapsules of isoniazid were prepared by phase separation coacervation process induced by non-solvent addition and using ethylcellulose (EC) as coating polymer. When polyisobutylene (PIB)--a protective colloid was present at sufficient concentration, film coated drug particles were formed. At 0-6% PIB concentration, the microcapsules were aggregated. Increase of colloid concentration produced microcapsules of less aggregation and higher drug content because coating became progressively thinner. PIB concentration also controlled the particle size and the release rate of drug from microcapsules. Wall thickness and EC loss were calculated from drug content. Microcapsules coated with EC were prepared with 7-9% PIB. Scanning Electron Microscopy was used to study the nature of aggregation and coating behaviour. The in vitro dissolution study confirmed the first order release pattern and also the Higuchi Matrix model.

Dysphagia caused by a fetus-in-fetu in a 27-year-old man.

Mechanical obstruction of the distal esophagus by a fetus-in-fetu is an extremely rare condition that has not been previously reported. We present the case of a 27-year-old man who presented with dysphagia caused by fetus-in-fetu contained within a retroperitoneal cystic cavity. The tumor, noticed since childhood, did not cause any symptoms until a year before presentation when symptoms of dysphagia developed. We propose including this entity in the differential diagnosis of a retroperitoneal mass.

Study on the diuretic activity of Strychnos potatorum Linn. seed extract in albino rats.

Methanol extract of Strychnos potatorum Linn. seeds (SPSE) was evaluated for its diuretic activity in Wistar albino rats. The SPSE was administered at the graded doses of 200, 400, and 600 mg/kg body weight. The parameters which were taken into account during the experimental on each rat were: total urine volume (corrected for water intake during the test period), body weight before and after the experiment, and the concentration of sodium, potassium, and chloride ions in urine. The total urine volumes of the SPSE (600 mg/kg)-treated rats were evaluated nearly two and half fold then compared with the control (saline treated) group. Excretion of cations (sodium and potassium ions) and anions (chloride ions) also increased significantly with respect to the control group. The diuretic effect was comparable with that of the standard drug Furosemide. The increase of cations in the urine on treatment with Strychnospotatorum seed extract (SPSE) was dose-dependent. This effect supports the use of the Strychnos potatorum seeds as a diuretic in folk remedies.