Indian Spices Enhance the Activities of Antibiotics against Human Pathogens Synergistically.

Background: Bacterial pathogens are considered aspredominant cause of human diseases throughout theworld. Until recently, antibiotics were considered aspromising agents against most bacterial pathogens butrecent reports suggest that there is growing resistance tocommonly used antibiotics creating a global healthcareproblem. Aim and Objectives: To investigate thesynergistic antibacterial potential of three differentantibiotics including Vancomycin, Clindamycin andCefotaxime with three popular Indian spices namelyCinnamomum zeylanicum (Dalchini), Trachysparmumammi (Ova) and Syzygium aromaticum (Clove) againsthuman pathogens Staphylococcus aureus,Enterococcus faecalis, Escherichia coli and Klebsiellapneumoniae. Material and Methods: Fourier-transformInfrared (FTIR) spectroscopy analysis was performedto detect molecular changes occurring while synergisticexposure of antibiotics and spices on pathogenicmicrobes. The addition of spices extracts showedenhanced activity of antibiotics against the pathogenshowever degree of antibiosis was varied according tobacterial species. Inhibition Zones (IZ) ranged from 0.0-34 mm. The highest IZ of 34.33 mm was found againstS. aureus where a combination of Cefotaxime and C.zeylanicum were applied. The synergy of spice extractswith antibiotics revealed an increase in the bactericidalactivity of standard antibiotics against pathogens. FTIRspectral analysis showed that, microorganisms showingresistance to antibiotics (Vancomycin and Clindamycin), alters important functional groups of antibioticsmight be resulting in decreased antimicrobialperformance. FTIR spectra's revealed common bands inantibiotics and spices such as nitroamines, aromaticphosphorus, benzene, bromide, carboxylic group,aliphatic esters, sulphonamides, primary alcohols,aliphatic ether, acid anhydride conjugate ring withketone and azo compounds, aromatic ethers, sulphonilchloride, sulphoamide etc. Interestingly, there wasincreased antimicrobial response for synergism whendecreased concentration of antibiotics and increasedconcentration of spice extracts were used. Conclusion:This investigation suggests that, spice extract could beused independently and in combination to elevate theperformance of antibiotics which addresses the issue ofdrug resistance in human pathogens.

Synchronous gastrointestinal stromal tumor of stomach and neuroendocrine tumor - A rare case report

Gastrointestinal stromal tumor (GIST) is the most common mesenchymal tumor occurring in the stomach with characteristic morphological features. Approximately 7–8% of all neuroendocrine tumors (NETs) are gastric NETs. However, simultaneous occurrence of both of them in stomach is a rare event. Here we present a rare case of synchronous GIST and NET in stomach of a 73 years old female patient. In our case, GIST showed Low risk category morphology as well as characteristic CD117 positivity proven by Immunohistochemistry. NET, our case, was well differentiated Grade I type showing positivity for synaptophysin and chromogranin A. Reporting of such rare cases is important for surgeons as well as pathologists to provide better patient management.

Analytical Method Development And Validation For Simultaneous Estimation Of Curcumin And Cyclosporine By Rp-Hplc

Objective: The present work was undertaken with an aim to develop and validate a rapid reverse-phase high-performance liquid chromatography (RP-HPLC) method for the estimation of curcumin and cyclosporine in the capsule dosage form. Methods: The RP-HPLC method for the simultaneous estimation of curcumin and cyclosporine was developed using Agilent (Infinity 1260) HPLC system and Eclipse XDB-C18 (4.6 x 150 mm i.d., 5µ) stationary phase. The optimized mobile phase comprised of acetonitrile: water: methanol (50: 10: 40 v/v/v) pumped at a flow rate of 0.5 ml/min. Separation of drugs was achieved in an isocratic mode and elution was monitored using PDA detector at 214 nm. The method was validated as per ICH-Q2R1 guidelines. Results: Retention time of the curcumin and cyclosporine were found to be 3.073 min and 6.373 min with the correlation coefficient (R2) of 0.9993 and 0.998 respectively. The response of curcumin and cyclosporine was found linear in the concentration range of 8-48 μg/ml and 4-24 μg/ml respectively. The percent recovery values were found in the range of 97-103% indicating satisfactory accuracy of the method. The percent relative standard deviation (% RSD) values for the precision study was less than 2 which suggest that the method is precise. Conclusion: The proposed method was found accurate, precise and specific for the determination of curcumin and cyclosporine in bulk as well as in capsule dosage form. Thus, the present method can be used for routine analysis and quality control of curcumin and cyclosporine in bulk and capsule dosage form.

Cytotoxicity of Mycobacterium indicus pranii on Mia-Pa-Ca2 cells

Background: MIP is a cultivable, non-pathogenic organism, which shares several antigens with Mycobacterium tuberculosis and Mycobacterium leprae. It has several proposed clinical applications. However, its cytotoxic effect on pancreatic cancer has not been documented. Hence, the study was conducted to investigate MIP induced cytotoxicity on Mia-Pa-Ca2 cells. To determine the cytotoxic potential of heat killed Mycobacterium indicus pranii (MIP) on pancreatic cancer cells in vitro along with gemcitabine & 5-fluorouracil (5-FU). Mitogen-activated protein kinase (MAPK) level was also studied post MIP treatment. Methods: Cytotoxic effect of MIP, gemcitabine and 5-FU on Mia-Pa-Ca2 cells was determined. We have analyzed extent of apoptosis using flow cytometry and changes in p38 levels, c-Jun N-terminal kinases (JNK) and extracellular signal­regulated kinase (ERK) using ELISA. Results: MIP not only exhibits cell cytotoxicity in dose dependent manner, but also enhances efficacy of gemcitabine and 5-FU when used in combination. Flow cytometry analyses reveals apoptosis of Mia-Pa-Ca2 cells post MIP treatment compared to untreated cells. MAPK pathway study using ELISA shows that p38 and JNK levels are suppressed while there is no change in ERK level. Conclusion: With these results we conclude that MIP is a cytotoxic agent. Cytotoxicity is exhibited by apoptosis. Combining MIP with gemcitabine and 5-FU shows synergistic effect (AU)

Laboratory diagnosis of malaria-various method and it’s comparision.

This is a comparative review of various techniques used for the detection of various malarial species, it’s specificity, sensitivity and availability at tertiary care center with its cost effectiveness.

Role of antioxidant therapy in management of type – 2 diabetes mellitus.

Introduction: Diabetes Mellitus is a heterogeneous group of disorders characterized by variable degrees of insulin resistance, impaired insulin secretion and increased glucose production. Free radical injury is important contributing factor for the development of Insulin resistance and impaired insulin secretion. Recently it has been suggested that glycation of antioxidant enzymes could alter the structure and function of antioxidant enzymes such that they are unable to detoxify free radicals. Intake of vitamin E and vitamin C, due to antioxidant property, is associated with reduced risk of development of Diabetes Mellitus Type 2. With this background, this study was planned to explore the role of antioxidant therapy in the management of Diabetes Mellitus Type 2. Objectives: (1) To demonstrate increased oxidative stress in newly diagnosed Type 2 Diabetes Mellitus patients by measuring antioxidant enzymes activities. (2) To study the effect of oral hypoglycaemic agents on oxidative stress in Type 2 Diabetes Mellitus patients. (3) To evaluate the effects of vitamin C, vitamin E and their combination in patients having Type 2 Diabetes Mellitus managed with oral hypoglycaemic agents. Materials and Methods: The study included two groups consisting of 60 euglycemic healthy subjects and 64 newly diagnosed Diabetes Mellitus Type 2 patients. After 3 months of treatment with oral hypoglycaemic drug, the second group was divided into 4 subgroups with 16 subjects in each subgroup and were treated with oral hypoglycaemic agent alone, oral hypoglycaemic agent + vitamin C, oral hypoglycaemic agent + vitamin E, and oral hypoglycaemic agent + vitamin C + vitamin E respectively for further 3 months. Results : Hyperglycemia in patients with Diabetes Mellitus Type 2 is associated with reduced antioxidant status (superoxide dismutase and catalase activities, and reduced glutathione level). Treatment with oral hypoglycaemic agent for 3 months produced euglycemia with partial but statistically significant elevation of catalase activity and reduced glutathione level in blood. Following additional antioxidant therapy with vitamin C and vitamin E produced further significant increase in reduced glutathione level, however fasting plasma glucose level and activities of superoxide dismutase and catalase enzymes were found to be statistically non-significant.Conclusion: Antioxidant therapy with vitamin C and vitamin E in addition to oral hypoglycaemic agent reduces oxidative stress in patients having Type 2 Diabetes Mellitus.

Concentration of nitrite in respirable particulate matter of ambient air in Vadodara, Gujarat, India.

Water extract of respirable particulate matter (RPM) was analyzed by Ion chromatography technique to investigate the presence of nitrite (NO2) as secondary aerosol in ambient environment. The nitrite particulates undergo photo hydroxyl radical reaction in environment produce nitrous acid, which reacts with metal and absorbs on RPM as water-soluble metal salt. The mean concentration of nitrite was 20.86 .g m-3 in ambient environment. Regression analysis showed that the relationship for respirable particulate matter and nitrite (RPM-NO2, R2=0.742) was positively significant. We are reporting the presence of nitrite as an aerosol in ambient environment.

Myotonic dystrophy in a patient of celiac disease: a new association?

Celiac disease (CD) has long been known to be associated with neurological and psychiatric manifestations; its in association with myotonic dystrophy however has not yet been reported. We report the case of a 27-year old female patient who presented to us with diarrhoea, weight loss, easy fatigability, irritability and alopecia of 8 months duration and was diagnosed to have celiac disease and put on gluten free diet. 8 weeks later she developed neurological symptoms and was found to have myotoni dystrophy in addition. At six month follow up patient had gained 5 kg, but the neurological symptoms remained the same. Treatment of neurological symptoms associated with gluten hypersensitivity depends on the type of neurological syndrome associated. Only exceptionally do these symptoms improve with gluten restriction and, in some patients, the neurological manifestations even progress despite resolution of both pathologic findings and intestinal symptoms.

Can anti-Helicobacter pylori and anti-CagA antibodies be used to select patients with dyspepsia for gastroscopy?

BACKGROUND: agA IgG antibody in sera may indicate presence of peptic ulcer disease among dyspeptic patients and therefore may be used as a serological marker to identify high risk patients for peptic ulcer who can be subjected to endoscopy. Present study was performed to identify association of CagA IgG antibody in patients with peptic ulcer. METHODS: Consecutive patients with dyspepsia were subjected to endoscopy and sera was collected from each. Rapid urease test in antral tissue collected from each patient by endoscopic biopsy was performed. Antral tissue was also examined histologically. IgG Antibody against H. Pylori and CagA IgG antibody was tested in each patients sera. RESULTS: Out of 82 patients with dyspepsia included in the study 28 had peptic ulcer. Of whom 26 were positive for anti IgG H. Pylori antibody. More than 80% patients with peptic ulcer patients had detectable anti Cag A antibody in contrast to 33% patients with non ulcer dyspepsia (P < 0.001). CONCLUSION: Anti-Cag A antibody may be used as a screening test in patients with dyspepsia to select high risk patients for peptic ulcer for upper gastrointestinal endoscopy.

A correlation of secondary aerosol (nitrate and sulfate) with respirable particulate matter (RPM) in ambient air at different traffic junctions of Vadodara city.

The correlation study of secondary aerosol (nitrate and sulfate) with RPM in ambient air at different traffic junctions of Vadodara city is reported. RPM was analyzed using Ion Chromatography technique and measured the level of nitrate and sulfate in ambient air. The correlation studies of these particulates with RPM have been established. The average concentration of sulfate and nitrate in ambient air was found 35.74 microg/m3 and 24.22 microg/m3, which ranged of 5.33-84.69 and 1.93-77.86 microg/m3 respectively. The correlation of RPM and SO4 (r = 0.813, P<0.01), RPM-NO3 (r = 0.5549, P<0.01) and SO4-NO3 (r = 0.6133, P<0.01) were found significant. The presence of sulfate and nitrate in RPM is 8.25% and 5.60% . The pH of water extract of RPM averaged 6.81, which ranged 6.17-7.28. Regression analysis result showed that the relationship between RPM-SO4 was significantly (R2=0.66215) correlated. This indicate that probably the secondary aerosols such as nitrate and sulfate in excess may cause irritation and increasing lung disease.

Celiac disease in osteoporotic Indians.

OBJECTIVE: 1) The aim of the study was to identify the atypical celiac disease (CD) in a cohort of symptomatic osteoporotic patients, younger than 55 years of age and 2) To study associated clinical and laboratory features and outcome with gluten-free diet. MATERIAL AND METHODS: We studied 33 patients (F:M = 28:5), mean age 29 years (range 15-52 years) with osteoporosis (WHO diagnostic criteria, T-score less than -2.5 on DEXA scan) from January 2000 - June 2002. Serological screening for CD was done by detecting circulating IgA antibodies to tissue transglutaminase by ELISA. Patients with presence of antibodies to transglutaminase were subjected to biopsy from the 2nd part of the duodenum by upper GI endoscopy. The biopsies were reported independently by two pathologists who were blinded for the serology report. Measurement of mucosal thickness, crypts and villi were done with an ocular micrometer. Other parameters like complete hemogram, serum iron, total iron binding capacity (TIBC), calcium profile, 25-OH-D, parathyroid hormone (PTH) were evaluated. Assessment of clinical and laboratory parameters was performed within 4-12 weeks of starting gluten-free diet (GFD). RESULTS: Thirteen patients had circulating IgA antibodies to transglutaminase. Intestinal biopsies were performed on 11 patients and were consistent with the diagnosis of CD (total villous atrophy--two, subtotal villous atrophy with crypt hyperplasia--nine). Patients with CD had significant anaemia when compared with non-CD osteoporotic patients. Other important observations in these 11 patients were low serum calcium and phosphorus, low 25-OH-D, high PTH. Significant improvement in clinical and laboratory parameters was noted in all patients within 6-12 weeks of starting GFD. CONCLUSION: Symptomatic osteoporotic patients (younger than 55 years of age) especially with associated anaemia should be investigated for CD. Simple measures like omission of wheat from diet (GFD) lead to significant improvement in symptoms within weeks.

Validity of self-report of recent opiate use in treatment setting.

Self-report validity of recent drug use among heroin abusers depends on many factors including the population being studied and the setting in which the study is carried out. This study was conducted by the treating physicians to assess the self-report validity of recent heroin use by heroin dependent patients in the outdoor setting using 'thin layer chromatography' (TLC) and two highly sensitive methods of urinalysis viz. 'gas liquid chromatography' (GLC) and 'high performance liquid chromatography' (HPLC). Out of seventy-six heroin dependent patients who entered the study, 64 provided urine sample on the same day. Patients' self-report about recent opiate use was found to have a moderate agreement with urinalysis report. However, it is important to validate it with urinalysis during the treatment process because a substantial proportion of patients fails to report recent opiate use. It is recommended that all drug dependence treatment centres should be equipped with a sensitive urinalysis facility. Otherwise, the outcome of the treatment process should be considered with caution.

Efficacy of low dose intradermal hepatitis B vaccine: results of a randomized trial among health care workers.

BACKGROUND: Hepatitis B virus (HBV) infection is an occupational health hazard among the healthcare workers. Vaccination against HBV has been established to be the most effective preventive strategy. The present study was designed to assess the efficacy of low dose intradermal HBV vaccine among the nursing staff in a tertiary care hospital setting. PATIENTS AND METHODS: Staff nurses working in our hospital were included in the study as vaccine recipients. Each staff nurse was tested for HBsAg and anti-HBs (commercial ELISA). Those who tested negative for both the above markers were randomized to receive either three doses of intramuscular (i.m.) HBV vaccine (20 micrograms m each dose) at 0, 1 and 6 month interval or three doses of intradermal HBV vaccine (2 micrograms m each dose) at similar intervals. Each vaccine recipient was tested for the presence of anti HBs (commercial ELISA) at the end of 1 month and 1 year after the last dose of the vaccine. The anti-HBs titres were also estimated simultaneously in them. RESULTS: Out of 153 staff nurses screened, 19 were either positive for HBsAg (n = 1) or anti HBs (n = 18). 96(72%) of the remaining 134 nurses agreed to receive HBV vaccine (i.m.--48, intradermal--48). At the end of 1 month after last dose of the vaccine, all vaccinees in both the group tested positive for anti-HBs. However the anti-HBs titres at 1 month were significantly higher among intramuscular vaccinees than the nurses receiving the vaccine through intra-dermal route (253 +/- 127.7 mIU/ml vs 151.3 +/- 92.8 mIU/ml, P < 0.001). Eighty four (85.5%) of these 96 vaccine recipient were available for evaluation of anti-HBs titre at the end of 1 year after the last dose of vaccine (1M group = 40, Intradermal group = 44). All the nurses continued to be positive for anti-HBs at the end of 1 year but the anti HBs-titre among i.m. vaccine recipient continued to remain at a significantly higher level than the similar titre among the intradermal vaccine recipients (256.4 +/- 124.7 mIU/ml vs 121.6 +/- 122.4 mIU/ml p < 0.001). CONCLUSION: Intradermal route for HBV vaccine had similar immunogenic efficacy as the conventional intramuscular route, but the dose required in the former route is one tenth of the intramuscular route. Therefore intradermal route may reduce the cost of HBV vaccine markedly.

Duodenal leiomyoma--a rare cause of haematemesis.

Leiomyoma of the duodenum is a rare tumour. Small intestinal tumours contributing to upper gastrointestinal bleed is still rare. They usually present with malena and anaemia, rarely hematemesis. We report a case of leiomyoma of duodenum diagnosed on endoscopic ultrasound that presented with massive haematemesis.

Comparative bioavailability study of a conventional and two controlled release oral formulations of Tegretol (carbamazepine)--200 mg.

OBJECTIVES: To assess the bioquivalence of carbamazepine (CBZ) controlled release formulation A (Tegretol CR, local) vs formulation B (Tegretol CR, Basel) and confirm their controlled release characteristics by comparing with conventional formulation (Tegretol). METHODS: A three-way randomized cross-over bioavailability study was carried out using CBZ 200 mg tablets of conventional and two controlled release formulations in twelve healthy volunteers. Coded plasma samples were analysed for levels of CBZ by HPLC method. RESULTS: The mean Cmax, Tmax, t1/2 and AUC for formulation A were: 1.67 +/- 0.26 mcg/mL, 24 +/- 0 hr, 47.8 +/- 9.7 hr and 136.7 +/- 25.4 mcg/ml. h; for formulation B were 1.41 +/- 0.31 mcg/mL, 25 +/- 8 hr, 46.9 +/- 7.9 and 119 +/- 32.3 mcg/ml.h and for conventional formulation were 2.43 +/- 3.6 mcg/mL, 9.5 +/- 7.4 hr, 44.6 +/- 9.8 hr and 178.8 +/- 41.9 mcg/ml.h respectively. The fluctuation in plasma concentration within 24 h (peak:trough) were 11.7 +/- 8.14% with conventional formulation as compared to 0% and 1.2 +/- 3.98% with formulation A and B respectively. The mean Tmax for both the controlled release formulations was not statistically significant. On the basis of 90% confidence interval, mean AUC and Cmax values obtained after controlled release formulation A, though statistically significant (P < 0.05) lie well within the prescribed limits of 80-120% as compared to formulation B. Thus both the controlled release formulations were bioequivalent. In comparison to conventional formulation, both controlled release formulations gave lower Cmax, lower AUCs, higher Tmax values, less fluctuation in CBZ plasma concentrations, reduction in ratio of Cmax/AUC values, thus demonstrating controlled release characteristics of the formulation. CONCLUSIONS: Based on the above mentioned parameters both controlled release formulations are bioequivalent and demonstrate controlled release characteristics.

Comparative study of platelet histamine and serotonin with their corresponding plasma oxidases in asthmatics with normals.

OBJECTIVE: Asthma is well controllable but non-curable disease. Exact pathophysiology involved is unresolved till today. Role of allergic hypersensitivity reaction in asthmatic on-set is well established. Present work is an effort to elucidate some basic points of unresolved pathophysiology of asthma taking platelets as marker. MATERIAL AND METHODS: A group of 52 normal human subjects in the age group of 20-60 years were studied for platelet histamine and serotonin levels and also for their plasma metabolising enzymes diamine oxidase (DAO) and monoamine oxidase (MAO). The data was collected for 79 asthmatic patients at different stages of asthma and accordingly were studied as four different groups of seventy nine asthmatics those were on regular treatment and were comfortable with drugs and were free from symptomatic attack formed gr. I; these (79) patients were followed-up during their symptomatic phase (gr. II) and same (79) patients immediately after their recovery from symptomatic stage studied as gr. III members. All the 79 asthmatic patients fall in gr. I, II and III in a serial manner i.e. all (n = 79) in each group. A separate group of thirty seven patients with known history of asthma but were symptom free and also off drugs for last 2-4 years formed gr. IV. RESULTS: Results showed mean platelet count in asthmatics at all four stages were in the normal range but were slightly low in comparison with normals. Both the enzymatic levels (DAO and MAO) in gr. I, II and III were significantly higher than normals but were same in the case of gr. IV patients. Low levels of platelet biogenic amines were observed in asthmatics (gr. I to gr. IV) than normals. CONCLUSIONS: Thus, study parameters showed significant difference in asthmatics and normals. Findings of the study have been utilized to understand unanswered hypersensitivity shown by the asthmatics over normal individuals (non-asthmatics).

Comparison of absorption rate and bioavailability of two brands of carbamazepine.

OBJECTIVE: To assess the bioavailability of carbamazepine from two brands of carbamazepine--Tegretol 200 and Zen-200. METHODS: A two-way randomised cross-over bioavailability of carbamazepine was carried out in twelve healthy male volunteers. Coded plasma samples were analysed for levels of carbamazepine by high performance liquid chromatography (HPLC) method. Tegretol 200 and Zen-200 were tested for in-vitro dissolution profiles. RESULTS: The mean Cmax, Tmax and t1/2a for Tegretol 200 were: 2.17 +/- 0.42 mcg/mL, 11.67 +/- 6.37 h and 2.72 +/- 1.87 h; for Zen-200 were 3.10 +/- 0.05 mcg/mL, 3.50 +/- 2.11 h and 0.76 +/- 0.76 h respectively. These values were statistically significant. However AUC (0-96 h) value of 150.16 +/- 27.13 mcg/ml.h after Zen-200 was not statistically significant as compared to 128.68 +/- 20.22 mcg/ml.h after Tegretol 200. The in-vitro dissolution profiles of the two formulations were dissimilar. The fluctuations in CBZ levels after Tegretol 200 was significantly less as compared to Zen-200. The absorption profile as judged by parameter 'A' was 50.44 +/- 10.95 for Tegretol 200 and 42.49 +/- 18.89 for Zen-200. CONCLUSION: Based on parameter 'A' and other pharmacokinetic parameters, the marketed generic carbamazepine product, Zen-200 is not bioequivalent to Tegretol 200.

Synthesis and anti-HIV activity of some non-nucleoside 2,3-disubstituted quinazoline derivatives (Part-V).

Several [2-phenyl-4(3H)-oxo-3-quinazolinylamino]-N-substituted- arylacetamides (1a-j) have been synthesized and tested at the National Cancer Institute, Bethesda, Maryland, USA, for their anti-HIV activity against susceptible human host cells (CEM cell line) over a wide range of concentrations (6.35 x 10(-8) to 2.00 x 10(-4) M). The highest protection observed is 45.67%. The structures of these compounds have been established on the basis of elemental analysis and spectral data.

Neuroleptic malignant syndrome: an experience of four cases.

We report four cases of neuroleptic malignant syndrome occurring after administration of a typical antipsychotic haloperidol and a newer atypical antipsychotic clozapine. The management of these patients is discussed.