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July-August 2016
Volume 48 | Issue 4
Page Nos. 347-471

Online since Wednesday, July 13, 2016

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EDITORIAL  

Fixed dose drug combinations: Issues and challenges in India Highly accessed article p. 347
Yogendra Kumar Gupta, Suganthi S Ramachandran
DOI:10.4103/0253-7613.186200  
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RESEARCH ARTICLES Top

Evaluation of CYP2C19, P2Y12, and ABCB1 polymorphisms and phenotypic response to clopidogrel in healthy Indian adults Highly accessed article p. 350
Kannan Sridharan, Rachna Kataria, Drishti Tolani, Shital Bendkhale, Nithya J Gogtay, Urmila M Thatte
DOI:10.4103/0253-7613.186191  
Introduction: CYP2C19 and P2Y12 polymorphisms have been claimed to alter the pharmacodynamic response to clopidogrel. ABCB1 polymorphism has been associated with the efflux of clopidogrel resulting in decreased bioavailability. Due to paucity of data from Indian population, the present study was undertaken to evaluate the association of genetic polymorphisms of CYP2C19, P2Y12, and ABCB1 with inhibition of platelet aggregation (IPA) by clopidogrel. Methods: Healthy adults (n = 90) of either gender were administered single dose of 300 mg clopidogrel. Baseline, 4 h postdose, and day 7 assessment of platelet aggregation and genotype of CYP2C19, P2Y12, and ABCB1 were carried out using standardized laboratory methods. The difference in the maximum platelet aggregation (MPA) between baseline and 4 h postdose was considered as delta-MPA (DMPA), and percentage change of MPA at 4 h from baseline was considered as IPA. Those with an IPA of <30% were considered as poor responders. Inferential statistics was applied to find out significant difference of these parameters between various groups of genetic polymorphisms. Results: Mean (standard deviation [SD]) of MPA (%) at baseline, 4 h postdose, and day 7 were 78 (5), 56 (16), and 71 (8), respectively. Similarly, mean (SD) of DMPA (%) and IPA (%) were 23 (17) and 29 (21), respectively. A total of 54/90 (60%) cases were found to be poor responders to clopidogrel. A wild genotype (*1/*1) of CYP2C19 was observed in 35 (40.2%), 42 (48.3%) had *1/*2, 2 (2.3%) individuals had *1/*3, and 8 (9.2%) had *2/*2 mutant genotypes. Although statistically not significant (P = 0.09), a trend was observed in having decreased inhibition values (both MPA and IPA) as we proceed from wild genotype (*1/*1) to mutant genotypes in the order of *1/*2, *1/*3, and *2/*2. Similarly, in P2Y12, a wild haplotype (H1/H1) was present in 77 (89.5%) and 9 (10.5%) individuals had H1/H2 type. A statistically significant difference in DMPA and IPA was observed with more IPA by clopidogrel in individuals with H2 haplotype. No association was observed between the carriers and noncarriers of mutant (T) allele of ABCB1. Conclusion: A trend of decrease in the IPA with CYP2C19 genotypes and an increase in the same with the H2 haplotype of P2Y12 following clopidogrel in Indian healthy adults were observed. Assessment of genetic polymorphisms of the same may aid in personalizing the therapy with clopidogrel.
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Platelet-to-lymphocyte ratio but not neutrophil-to-lymphocyte ratio predicts high on-treatment platelet reactivity in clopidogrel-treated patients with acute coronary syndrome p. 355
Edem Efe, Ibrahim Kocayigit, Pabuccu Mustafa Turker, Kucukukur Murat, Alpaslan Erkan, Tas Sedat, Cil Alper, Aksoy Murat Necati, Vural Mustafa Gokhan, Akdeniz Bahri
DOI:10.4103/0253-7613.186205  
Objectives: Dual antiplatelet therapy (DAPT), consisting of clopidogrel and aspirin, is the main-stay treatment of acute coronary syndromes (ACS). However, major adverse cardiovascular events may occur even in patients undergoing DAPT, and this has been related to the variable pharmacodynamic efficacy of these drugs, especially clopidogrel. Platelet-to-lymphocyte ratio (PLR) and neutrophil-to-lymphocyte ratio (NLR) are novel inflammatory markers for cardiovascular risk stratification, which may reflect an inflammatory state and thus high on-treatment platelet reactivity (HPR). Methods: We investigated the usefulness of PLR and NLR in predicting HPR in clopidogrel-treated patients with ACS. A total of 244 patients were enrolled in this study, and 43 of them were nonresponsive to clopidogrel. Results: Logistic regression analysis indicated that PLR was significantly associated with HPR (P < 0.001). Using a cutoff level of 331, PLR predicted HPR with a sensitivity of 73% and a specificity of 69% (odds ratio: 376.15, 95% confidence interval = 37.813–3741.728 P< 0.001, receiver operating characteristic curve: 0.885). Conclusions: We suggest that more attention should be paid to the PLR values of these patients on admission to identify individuals who may not benefit from clopidogrel during the course of ACS.
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Drug dispensing practices at pharmacies in Bengaluru: A cross-sectional study p. 360
R Soumya, Vijayalakshmi Devarashetty, CR Jayanthi, M Sushma
DOI:10.4103/0253-7613.186204  
Objectives: Pharmacists are one of the crucial focal points for health care in the community. They have tremendous outreach to the public as pharmacies are often the first-port-of-call. With the increase of ready-to-use drugs, the main health-related activity of a pharmacist today is to assure the quality of dispensing, a key element to promote rational medicine use. Materials and Methods: A cross-sectional study of 200 pharmacies, 100 each in various residential (R) and commercial (C) areas of Bengaluru, was conducted using a prevalidated questionnaire administered to the chief pharmacist or the person-in-charge by the investigators. Results: Dispensing without prescription at pharmacies was 45% of the total dispensing encounters and significantly higher (c2 = 15.2, P< 0.001, df = 1) in pharmacies of residential areas (46.64%) as compared to commercial areas (43.64%). Analgesics were the most commonly dispensed drugs (90%) without prescription. Only 31% insisted on dispensing full course of antibiotics prescribed and 19% checked for completeness of prescription before dispensing. Although 97% of the pharmacies had a refrigerator, 31% of these did not have power back-up. Only about 50% of the pharmacists were aware of Schedule H. Conclusion: This study shows a high proportion of dispensing encounters without prescription, a higher rate of older prescription refills, many irregularities in medication counseling and unsatisfactory storage practices. It also revealed that about half of the pharmacists were unaware of Schedule H and majority of them about current regulations. Hence, regulatory enforcement and educational campaigns are a prerequisite to improve dispenser's knowledge and dispensing practices.
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Knowledge and perceptions on antibiotic use and resistance among high school students and teachers in New Delhi, India: A qualitative study p. 365
Anita Kotwani, Chand Wattal, PC Joshi, Kathleen Holloway
DOI:10.4103/0253-7613.186208  
Objective: To explore the perceptions and knowledge of school teachers and students about antibiotic use, resistance, and suggestions for practical interventions for the rational use of antibiotics. Methodology: Five focus group discussions (FGDs) with high school students (Class: 9–11) and five with teachers were conducted in two private and three public schools (one teacher and one student FGD per school) in five municipal wards of Delhi. Qualitative data on antibiotic knowledge, resistance, and behaviors with respect to antibiotics use were collected. There were 4–8 persons per teacher FGD and 15–20 persons per student FGD. FGDs were analyzed using “thematic analyses.” Results: Students had poor knowledge regarding antibiotics and antibiotic resistance, while only some teachers had a basic understanding. Four broad themes needing attention emerged: definition of antibiotic and antibiotic resistance, antibiotic use behavior, doctor–patient relationship, and interventional strategies suggested to curtail the misuse of antibiotics and to spread awareness. In order to tackle these problems, both groups suggested a multipronged approach including robust public awareness campaigns also involving schools, better doctor–patient relationships, and stronger regulations. Conclusions: Although students and teachers exhibited poor knowledge about antibiotic use and resistance, they were keen to learn about these issues. School education programs and public education could be used to shape correct perceptions about antibiotic use among all stakeholders including children. This may help in the containment of antibiotic resistance and thus preservation of antibiotics for future generations.
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Role of antioxidant property of carvedilol in mild to moderate hypertensive patients: A prospective open-label study p. 372
Saleh Ayashi, Ahmad Reza Assareh, Mohammad Taha Jalali, Samaneh Olapour, Hamid Yaghooti
DOI:10.4103/0253-7613.186206  
Objective: Carvedilol is a nonselective third generation β-blocker that does not display the negative effects of traditional β-blockers. Regarding the antioxidant, anti-inflammatory and distinct metabolic properties of carvedilol which are similar to that of high-density lipoprotein (HDL) and paraoxonase 1 (PON1), the present study intends to investigate the effects of carvedilol treatment on malondialdehyde (MDA) and soluble lectin-like ox-low-density lipoprotein (LDL) receptor (sLOX-1) as markers of oxidative stress in association to lipid profiles, apolipoproteins (apo), and PON1 activity in hypertensive patients. Patients and Methods: This clinical trial study was performed on forty patients with mild to moderate essential hypertension. Subjects were studied before and after 2 months treatment with carvedilol, 25 mg daily. Lipids and lipoproteins were measured using a biochemistry analyzer. PON and arylesterase activity were assayed using paraoxon and phenyl acetate as substrates, respectively. MDA was quantified using a chemical colorimetric assay. ELISA was used to measure sLOX-1. Results: Our results showed that carvedilol treatment decreased systolic and diastolic blood pressure as much as forty and 16 mmHg, respectively (P < 0.001). It also increased HDL, total cholesterol, and serum PON1 activity (P < 0.05), but the levels of triglyceride, LDL, apo A-I, and apo B did not significantly change. There was an inverse correlation between serum PON1 activity and serum MDA. Conclusion: This study confirmed the antihypertensive effect of the drug and its beneficial metabolic effects through augmenting HDL and PON1 activity. We propose that the antioxidant effects of carvedilol can be partially attributed to increased PON-1 activity.
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Prescribing pattern of analgesics in orthopedic in-patient department at tertiary care hospital in Guwahati, Assam, Northeast India p. 377
Dwijen Kumar Choudhury, Babul Kumar Bezbaruah
DOI:10.4103/0253-7613.186207  
Objectives: The aim of this study is to evaluate the prescribing pattern of analgesics and analyze the rational use of analgesic in orthopedic in-patient department of tertiary care teaching hospital, Guwahati, Assam. Subjects and Methods: An observational and cross-sectional study was carried out for 1 month from April to May 2014. Collected data included age, sex, diagnosis and line of management during the study. The generic name and the average cost of treatment per patient were evaluated using Indian Drug Review, 2014. The prescribed drugs were assessed with respective National Model List of Essential Medicines (NLEM), 2011 and the rationality of prescriptions was determined using the World Health Organization indicators of drug utilization. The patients' details were recorded in a predeigned data collection form and results were analyzed by descriptive statistics. Results: Out of 200 patients, 123 were male and 77 were female. The average number of analgesic per prescription was 1.46. In this study, 55.5% of patients had received single analgesic. Diclofenac was the most commonly prescribed analgesic (43.49%). During hospitalization, majority of the patients have received parenteral preparation. Gastroprotective agents and antimicrobials were frequently prescribed along with analgesics. Out of 292 analgesics prescribed, 183 (62.67%) were from the NLEM, India. Furthermore, 176 (57.19%) analgesics were prescribed by generic name. The average cost of treatment per patient was 2151.72 INR. Utilization of analgesic in terms of defined daily dose/100 bed-days was 104.01. Conclusion: The percentages of analgesics prescribing from NLEM and the use of analgesic by generic name were found satisfactory. Regular educational interventions to improve prescribing practices among physicians at different levels may further promote rational prescribing.
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Comparative evaluation of nephroprotective potential of resveratrol and piperine on nephrotic BALB/c mice p. 382
Rahul Motiram Kakalij, B Dinesh Kumar, Prakash Vamanrao Diwan
DOI:10.4103/0253-7613.186215  
Objective: The objective of this study was to evaluate the nephroprotective potential of resveratrol and piperine at same dose on cationic bovine serum albumin (cBSA) induced immune complex glomerulonephritis (ICGN) in BALB/c mice. Materials and Methods: Female BALB/c mice were divided into five groups. Group I served as normal control (complete Freund's adjuvant + Saline). Two weeks later, Groups II, III, IV, and V were administered cBSA (13 mg/kg) via the caudal vein 3 times/week every alternative day for 6 weeks to induce ICGN. Simultaneously, from the 3rd week, Groups III, IV were treated with resveratrol and piperine up to 6 weeks. Group V was treated with methylprednisolone considered as a reference standard. Results: There was a significant decrease in albuminuria, serum creatinine, and blood urea nitrogen in Group IV animals when compared with Group III. In addition, Group III and IV have comparable results with cBSA treated animals. Concurrently, same groups showed significantly comparable variance in antioxidant enzymes, phagocytic index, and neutrophil adhesion assay. Group IV found to be more significant in IgG1 reduction than Group III. Conclusion: The findings of this study well-demonstrated that piperine has potential immunomodulatory and anti-inflammatory activity than resveratrol; therefore, piperine needs special attention in autoimmunity and inflammation research.
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B1-kinin receptors modulate Mesobuthus tamulus venom-induced vasosensory reflex responses in anesthetized rats p. 388
Sanjeev K Singh, Shripad B Deshpande
DOI:10.4103/0253-7613.186202  
Objective: Intra-arterial injection of Mesobuthus tamulus (BT) venom produces reflex vasosensory responses modulating cardiorespiratory parameters in albino rats. The present study was conducted to understand the role of kinin receptors in modulating vasosensory reflexes evoked by BT venom. Materials and Methods: In urethane-anesthetized rats, tracheostomy was performed to keep the airway patent. The femoral artery was cannulated proximally, as well as distally, to record the blood pressure (BP) and to inject the chemicals, respectively. Electrocardiographic and respiratory excursions were recorded to compute the heart rate (HR) and respiratory rate (RR). A group of animals was pretreated with saline/kinin receptor antagonists intra-arterially (B1/B2 receptor antagonists) before the injection of venom. Results: After intra-arterial injection of BT venom (1 mg/kg), there was an immediate increase in RR, which reached to 40% within 30 s, followed by a decrease of 40%. Further, there was sustained increase in RR (50%) up to 60 min. The BP started to increase at 40 s, peaking at 5 min (50%), and remained above the initial level up to 60 min. The bradycardiac response started after 5 min which peaked (50% of initial) at 25 min and remained at that level up to 60 min. In B1 receptor antagonist (des-Arg) pretreated animals, venom-induced cardiovascular responses were attenuated (by 20–25% in mean arterial pressure and HR) significantly but not in B2 receptor antagonist (Hoe-140) pretreated animals. Either of the antagonists failed to alter the RR responses. Conclusions: BT venom-induced vasosensory reflex responses modulating cardiovascular parameters are mediated via B1-kinin receptors in anesthetized rats.
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Role of fosaprepitant, a neurokinin Type 1 receptor antagonist, in morphine-induced antinociception in rats p. 394
Pranav Prasoon, Shivani Gupta, Rahul Kumar, Mayank Gautam, Saroj Kaler, Subrata Basu Ray
DOI:10.4103/0253-7613.186198  
Objectives: Opioids such as morphine form the cornerstone in the treatment of moderate to severe pain. However, opioids also produce serious side effects such as tolerance. Fosaprepitant is a substance P (SP) receptor antagonist, which is used for treating chemotherapy-induced nausea and vomiting. SP is an important neuropeptide mediating transmission of pain at the spinal level. Thus, it was hypothesized that combining morphine with fosaprepitant would increase the antinociceptive effect of morphine. The objectives were to evaluate the effect of fosaprepitant on morphine-induced antinociception in rats and to investigate its mechanism of action. Methods: Sprague-Dawley rats were injected with morphine (10 mg/kg twice daily) and/or fosaprepitant (30 mg/kg once daily) for 7 days. Pain threshold was assessed by the hot plate test. Expression of SP and calcitonin gene-related peptide (CGRP) in the spinal cords of these rats was evaluated by immunohistochemistry. Results: Morphine administration resulted in an antinociceptive effect compared to the control group (day 1 and to a lesser extent on day 4). The decreased antinociception despite continued morphine treatment indicated development of tolerance. Co-administration of fosaprepitant attenuated tolerance to morphine (days 1 and 3) and increased the antinociceptive effect compared to control group (days 1–4). Expression of SP was increased in the morphine + fosaprepitant group. Conclusions: The results show that fosaprepitant attenuates the development of tolerance to morphine and thereby, increases the antinociceptive effect. This is likely linked to decreased release of SP from presynaptic terminals.
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Acidosis reduces the function and expression of α1D-adrenoceptor in superior mesenteric artery of Capra hircus p. 399
Ipsita Mohanty, Sujit Suklabaidya, Subas Chandra Parija
DOI:10.4103/0253-7613.186199  
Objective: The objective of this study was to characterize the α1-adrenoceptor (α1-AR) subtypes and evaluate the effect of acidosis on α1-AR function and expression in goat superior mesenteric artery (GSMA). Materials and Methods: GSMA rings were mounted in a thermostatically controlled (37.0°C ± 0.5°C) organ bath containing 20 ml of modified Krebs-Henseleit solution, maintained at pHoof 7.4, 6.8, 6.0, 5.5, 5.0, and 4.5. Noradrenaline (NA)- and phenylephrine (PE)-induced contractile response was elicited in the absence or presence of endothelium and prazosin at pHo of 7.4, 6.0, and 5.0. The responses were recorded isometrically by an automatic organ bath connected to PowerLab and analyzed using Labchart 7.1.3 software. Expression of α1D-AR was compared at physiological and acidic pHousing reverse transcription-polymerase chain reaction (RT-PCR). Results: NA- and PE-induced contractile responses were attenuated proportionately with a decrease in extracellular pH (pHo), i.e. 7.4 → 6.8 → 6.0 → 5.5 → 5.0 → 4.5. Endothelium denudation increased the contractile response at both normal and acidic pHo. Prazosin (1 nM, 10 nM, and 0.1 μM) inhibited the NA- and PE-induced contractile response at pHo7.4 and the blocking effect of prazosin was potentiated at pHoof 6.0 and 5.0. RT-PCR analysis for α1D-AR in GSMA showed that the mRNA expression of α1D-AR was decreased under acidic pHoas compared to physiological pHo. Conclusion: (i) Adrenergic receptor mediates vasoconstriction in GSMA under normal physiological pHo, and α1Dis the possible subtype involved in this event (ii) acidosis attenuates the vasocontractile response due to reduced function and expression of α1D-AR and also increased the release of endothelial-relaxing factors.
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Evaluation of effects of melatonin and caffeic acid phenethyl ester on acute potassium dichromate toxicity and genotoxicity in rats p. 407
Mujgan Cengiz, Nurnisa Oya Alansal, Matem Tuncdemir, Gamze Tanriverdi, Burcu Bayoglu
DOI:10.4103/0253-7613.186213  
Objective: The aim of this study is to investigate the possible protective effects of melatonin and caffeic acid phenethyl ester (CAPE) on potassium dichromate (K2Cr2O7)-induced nephrotoxicity and genotoxicity. Methods: A total of 40 Wistar albino rats were divided into five groups: control, K2Cr2O7(K2Cr2O7 15 mg/kg, one dose, i.p.), K2Cr2O7+ melatonin, K2Cr2O7+ CAPE, and K2Cr2O7+ melatonin + CAPE. Urine and blood samples were collected from rats before scarification. One kidney was collected for histopathological studies, and the other was stored at −80°C for further determination of catalase (CAT), superoxide dismutase (SOD), malondialdehyde (MDA), glutathione (GSH), glutathione S-transferase (GST), and glutathione reductase (GR) levels with spectrophotometric method. Comet assay was used to evaluate the genotoxicity. Results: We observed a significant amelioration in genotoxicity by melatonin and simultaneous melatonin + CAPE treatment compared to K2Cr2O7group (p1, p2< 0.05). SOD, CAT, GSH, GST, and MDA levels did not change when compared with controls. When K2Cr2O7applied group was treated with melatonin and CAPE, neither melatonin nor CAPE made any changes in kidney GSH, GST, SOD, and MDA levels (P > 0.05). We noted that treatment with CAPE and melatonin + CAPE together caused a significant decrease in renal tissue damage, an upregulation in the kidney CAT levels (P < 0.05) and a slight healing at GR levels when compared with the K2Cr2O7group. Conclusion: Our results revealed, CAPE and melatonin may have protective effects on K2Cr2O7induced nephrotoxicity and cellular damage in rats.
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Immunomodulatory activity of aqueous extract of Nyctanthes arbor-tristis flowers with particular reference to splenocytes proliferation and cytokines induction p. 412
Chandrabhan Kumar Bharshiv, Satish Kumar Garg, AK Bhatia
DOI:10.4103/0253-7613.186210  
Objectives: To investigate the immunomodulatory activity of aqueous extract of Nyctanthes arbor- tristis flowers (NAFE) with particular reference to splenocytes proliferation and induction of cytokines. Materials and Methods: Antibody titer was determined by tube agglutination and indirect ELISA assay in four groups of mice-control, antigen alone, and NAFE-treated (400 and 800 mg/kg for 21 days) after immunization with Salmonella antigen while cellular immunity was studied in three groups of rats (control and NAFE-treated - 400 and 800 mg/kg) following DNCB application. Splenocytes from untreated and NAFE-treated rats were stimulated using concanavalin-A (Con-A) and optical density (OD) and stimulation index were determined. Splenocytes from control rats were also treated in vitro with NAFE (50–1600 μg/ml) and Con-A to determine the effect on splenocytes proliferation. Interleukin-2 (IL-2) and IL-6 levels in splenocytes supernatant from control and NAFE-treated rats and following in vitro treatment of splenocytes with NAFE (50–1600 μg/ml) were determined using ELISA kits. Results: Marked to a significant increase in antibody titer by both the methods in NAFE-treated mice and a significant increase in skin thickness in rats after challenge with DNCB, respectively suggested humoral and cell-mediated immunostimulant potential of NAFE. Significant increase in OD and stimulation index following e x vivo and in vitro exposure of splenocytes and sensitization with Con-A and significant elevation in IL-2 and IL-6 levels in splenocytes supernantant was also observed after their ex vivo and in vitro exposure to NAFE. Conclusion: Humoral and cell-mediated immunostimulant activity of NAFE seems to be mediated through splenocytes proliferation and increased production of cytokines, especially IL-2 and IL-6.
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Phytochemicals, antioxidant, and anthelmintic activity of selected traditional wild edible plants of lower Assam p. 418
Ananta Swargiary, Abhijita Daimari, Manita Daimari, Noymi Basumatary, Ezekiel Narzary
DOI:10.4103/0253-7613.186212  
Objective: Clerodendrum viscosum, Eryngium foetidum, Lippia javanica, and Murraya koenigii are one among the common wild edible plants in Northeast India which are also used as antidiabetic, stomach-ache relieving drugs, etc., The present study was aimed to reveal the phytochemical, antioxidant, and anthelmintic activity of the plants. Materials and Methods: The antioxidant capacity of methanolic extract of plants was studied by 1,1-diphenyl-2-picrylhydrazyl (DPPH), ferric reducing antioxidant power, TBARS, and total antioxidant activity (TAA). Total phenolics, flavonoids, Vitamin C, carbohydrate, and protein are also estimated following standard protocols. Anthelmintic activity of the extracts has also been studied in vitro against trematode parasites. Results: The result showed that the methanolic extracts of plants possess a substantial quantity of alkaloids, phenolics, flavonoids, proteins, carbohydrates, and Vitamin C. Phenolics, flavonoids, and Vitamin C contents were found higher in C. viscosum followed by M. koenigii, L. javanica, and E. foetidum. The in vitro antioxidant assays revealed substantial free radical scavenging property in all the plants. TAA increased in the order C. viscosum > M. koenigii > L. javanica > E. foetidum. Similarly, C. viscosum displayed a better antioxidant capacity with IC50values 29.74 ± 3.63 μg and 148.77 ± 18.38 μg for DPPH and thiobarbituric acid reactive species, respectively. In addition, the plant extracts also showed good anthelmintic activity against Paramphistomum sp. Time taken for paralysis and death were 0:56 ± 0:09 h and 1:35 ± 0:07 h for L. javanica at 50 mg/mL concentration. Conclusion: The study therefore suggests the importance of tested plants as a natural source of free radical scavenger and plausible veterinary uses.
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Effect of nitrergic system on colonic motility in a rat model of irritable bowel syndrome p. 424
Tijen Kaya Temiz, Omer Demir, Fatma Simsek, Yusuf Cem Kaplan, Selen Bahceci, Baris Karadas, Asli Celik, Gokhan Koyluoglu
DOI:10.4103/0253-7613.186189  
Aim: The aim of this study is to investigate whether nitric oxide (NO)-mediated colonic motility was altered in rat irritable bowel syndrome (IBS) model, using different isoforms of NO-synthase (NOS) inhibitors. Materials and Methods: The animal model of IBS-like visceral hypersensitivity was induced by intra-colonic infusion of 0.5% acetic acid (AA) in saline once daily from postnatal days 8 to 21. Control animals received saline instead of AA. Experiments were performed at the end of 8 weeks. Distal colon tissues were resected and direct effects of different NOS inhibitors; N-omega-nitro-L-arginine methyl ester hydrochloride, (L-NAME), ARL-17477 dihydrochloride hydrate (ARL 17477), N-[3-(Aminomethyl) phenyl] methyl]-ethanimidamidedihydrochloride (1400 W), and N5-(1-Iminoethyl)-L-ornithine dihydrochloride (L-NIO) were evaluated concentration-dependently in vitro tissue bath. Besides, morphology of both groups was assessed with hematoxylin and eosin (H and E) staining and the impact of NO antibodies was determined using the immunohistochemical method. Results: The mean pressure values of spontaneous contractions and KCL (80 mmol/L) responses of distal colonic segments were similar in normal and IBS rats. L-NAME and ARL-17477 significantly increased the mean pressure of spontaneous colonic contractions in normal rats versus own base values (P < 0.05), but this increase did not significantly different when compared to IBS rats. In H and E staining, there was no difference with regard to morphology between two groups. Neuronal NOS (nNOS) immunoreactivity was found to be significantly decreased in IBS when compared to control groups (P < 0.05). Conclusion: L-NAME and ARL-17477 mediated mean pressure values were found to be slightly decreased in IBS rats. These findings may be related to a decrease in nNOS level in IBS.
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Evaluation of antioxidant effect of Nerium indicum in anxious rats p. 430
Deepak Suresh Mohale, Alok Shiomurthi Tripathi, Abhijit V Shrirao, Amol G Jawarkar, Anil V Chandewar
DOI:10.4103/0253-7613.186201  
Aim: The aim of this study was to analyze the ethyl acetate extract of Nerium indicum (NIE) flower for its antioxidant effect in anxious Sprague–Dawley rats. Materials and Methods: Animals were divided into six groups (n = 6) and treated with 200 mg/kg and 400 mg/kg p.o. of NIE for 21 days to assess its preventive and curative effects. Anxiety was induced by isolating animals socially for 21 days. Elevated plus maze (EPM) and light and dark model were used for measuring anxiety in animals. Oxidative stress parameters such as lipid peroxidation (LPO), superoxide dismutase (SOD), catalase (CAT), and reduced glutathione (GSH) in blood and brain tissue homogenate were monitored after 21 days of social isolation in animals. Results: Rats were treated with NIE 200 mg/kg and 400 mg/kg p.o. Both the treatments showed a significant (P < 0.001) increase in the number of open arm entries and time spent in open arm in EPM when compared with the negative control. Results also demonstrated that there was a significant (P < 0.001) increase in the number of lightbox entries and time spent in light box in light and dark model when compared with negative control. There was a significant (P < 0.001) improvement in endogenous anti-oxidants such as SOD, CAT, reduced GSH, and decreased levels of LPO in blood and brain tissue when compared with the negative control. Conclusion: The present study suggests the role of NIE in the treatment of anxiety, possibly by modulating the oxidative stress.
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Effect of hydro-alcoholic extract of Vernonia cinerea Less. against ethylene glycol-induced urolithiasis in rats p. 434
Ravindra D Hiremath, Sunil S Jalalpure
DOI:10.4103/0253-7613.186211  
Objective: Aim of this study is to evaluate antiurolithiatic potential of whole plant hydro-alcoholic (30:70) extract of Vernonia cinerea Less. in accordance to its claims made in ancient literature and also being one of the ingredients of cystone, a marketed formulation widely used in the management of urolithiasis. Materials and Methods: To induce urolithiasis, 0.75% v/v ethylene glycol was administered orally for 14 days. The curative dose of 400 mg/kg b.w. and preventive doses of 100, 200, and 400 mg/kg b.w. were administered from 15th to 28th and 1st to 28 days, respectively. Cystone 750 mg/kg b.w. was selected as the reference standard for both curative and preventive doses. On 28th day, urinate of 24 h was collected and subjected for estimation of calcium, oxalate, and phosphates. Serum biochemical and kidney homogenate analysis was done for determination of renal oxalate contents. Results: The diseased Group II showed marked increase (P < 0.001 vs. normal Group I) in levels of urine calcium, oxalate, and phosphate. Serum creatinine, urea, and uric acid levels were also increased. Histopathological studies of kidney sections revealed significant changes. Treatment with hydro-alcoholic extract of V. cinerea showed significant (P < 0.01 vs. calculi-induced Group II) dose-dependent activity. A progressive increase in urine output, body weight, and decline in concentrations of stone-forming components such as calcium, oxalates, and phosphates was observed. Conclusion: It can be inferred that V. cinerea Less. is effective in ethylene glycol-induced urolithiasis and may have a potential in preventing and curing urolithiasis.
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SHORT COMMUNICATIONS Top

Anti-inflammatory activity of fenugreek (Trigonella foenum-graecum Linn) seed petroleum ether extract p. 441
Kilambi Pundarikakshudu, Deepak H Shah, Aashish H Panchal, Gordhanbhai C Bhavsar
DOI:10.4103/0253-7613.186195  
Objectives: The aim of the present work was to study the anti-inflammatory and anti-arthritic activities of petroleum ether extract of fenugreek seeds. Materials and Methods: Fenugreek seed powder was extracted in petroleum ether by cold maceration. This fenugreek seed petroleum ether extract (FSPEE) was analyzed by gas–liquid chromatography (GLC) and tested on rats against carrageenan and formaldehyde-induced paw edema, complete Freund's adjuvant (CFA)-induced arthritis and cotton pellet-induced granuloma. Changes in serum glutamic oxaloacetic tansaminase (SGOT), serum glutamate-pyruvate transaminase (SGPT), and alkaline phosphatase (ALP) activities in liver and serum were also studied in cotton pellet-induced arthritic rats. Data were analyzed by Student's t-test. P <0.05 was considered statistically significant. Results: GLC of FSPEE showed oleic (33.61%), linoleic (40.37%), and linolenic (12.51%) acids. With 0.5 mL/kg FSPEE treatment, there was 37% (P < 0.05) and 85% (P < 0.05) reduction in inflammation of the paw in carrageenan and formaldehyde-induced paw edema. In CFA-induced arthritis, a biphasic increase in paw volume followed by decrease was seen. There was 42.5% (P < 0.01) reduction in the weight of cotton pellets and significant (P < 0.01) reductions in the elevated SGPT and ALP activities in serum and liver of FSPEE (0.5 mL/kg) treated rats. Conclusion: Thus, petroleum ether extract of fenugreek seeds has significant anti-inflammatory and anti-arthritic activities which are due to the presence of linolenic and linoleic acids.
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Methanolic extract of Anthocephalus cadamba induces apoptosis in Ehrlich ascites carcinoma cells in experimental mice p. 445
Narayan Dolai, Aminul Islam, Pallab Kanti Haldar
DOI:10.4103/0253-7613.186190  
Objective: Anthocephalus cadamba (Roxb.) Miq. (Family: Rubiaceae), a folk medicine commonly known as “Kadam” in Bengali, has been used for the treatment of tumor. The methanolic extract of A. cadamba (MEAC) showing antitumor activity on Ehrlich ascites carcinoma (EAC) cells treated mice was already reported. This study was designed to study the apoptosis-inducing property of MEAC and its mechanism in EAC cells in mice. Materials and Methods: Apoptogenic morphology was determined by fluorescent DNA-binding double staining method using dyes acridine orange (AO)/ethidium bromide (EB). Comet assay was estimated to check the DNA damage. Flow cytometry (fluorescence-activated cell sorting [FACS]) was used to detect the apoptotic rate quantitatively by double labeling techniques using annexin V FITC/propidium iodide staining. Apoptotic protein expression was done using Western blotting assay method. Statistical Analysis: Results are expressed as mean ± standard deviation. Statistical analysis was performed using ANOVA followed by Dunnett's post hoc test of GraphPad Prism software. * P < 0.05, ** P < 0.01 and *** P < 0.001 were considered statistically significant. Results: Apoptosis-inducing effect of MEAC on EAC cells was confirmed from AO/EB staining and FACS analysis. MEAC treatment showed dose-dependent induction of DNA damage. Apoptosis was induced by increasing the expression of multiple downstream factors such as pro-apoptotic protein p53 and p21 in EAC. Bax was up-regulated and anti-apoptotic protein Bcl-2 was down-regulated resulting in decrease of the Bcl-2/Bax ratio by MEAC treatment. Conclusion: Experimental results revealed that MEAC induces apoptosis by modulating the expression of some pro-apoptotic and anti-apoptotic proteins in EAC and thus exerts its anti-tumor activity.
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Assessment of anxiolytic effect of nerolidol in mice p. 450
Rajesh Kumar Goel, Dilpreet Kaur, Priyanka Pahwa
DOI:10.4103/0253-7613.186188  
Aim and Objectives: The present study was to assess the anxiolytic effect of nerolidol in mice. Materials and Methods: The anxiolytic activity was examined using the elevated plus maze (EPM) and open field test (OFT), and motor coordination by rotarod test. Thirty Swiss albino mice were divided into five groups of six mice each. Group 1 received vehicle control (normal saline); Group 2 received diazepam (1 mg/kg); Groups 3, 4, and 5 received nerolidol 12.5, 25, and 50 mg/kg, respectively. Results: Nerolidol (12.5, 25, and 50 mg/kg) significantly (P < 0.05) increased the time spent and a number of entries in open arm as compared to vehicle control in EPM test. In OFT, the nerolidol showed a significant (P < 0.05) increase in number of rearings and time spent in center and periphery, suggesting exploratory behavior of animals. Furthermore, nerolidol did not alter the fall down latency in rotarod test. Conclusion: Our findings indicated that nerolidol exerts an anxiolytic effect without altering the motor coordination.
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DRUG WATCH Top

Melasma: A rare adverse effect of clomipramine p. 453
Sujita Kumar Kar
DOI:10.4103/0253-7613.186203  
Melasma is a hyperpigmented dermatological condition common in females. Drugs such as steroids, cosmetics, and photosensitizing agents are known to cause melasma. We report here a case of an adult male with obsessive-compulsive disorder, receiving clomipramine, who developed melasma.
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A clinically suspected case of Anaphylactoid reaction to vitamin K injection in a child – a case report and review of literature p. 455
Bharat Choudhary, Sweta Kumari, Bhavna Dhingra, Ratinder Jhaj
DOI:10.4103/0253-7613.186216  
Vitamin K is commonly indicated in pediatric patients with coagulopathies. Its commercial preparations are available in the market since long. Minor complications are very well-known, but life-threatening events are rarely reported in children. We present a case of 8-year-old child who developed life-threatening event following intravenous Vitamin K injection. She was survived after cardiopulmonary resuscitation. The reaction was most probably due to anaphylactoid reaction to Vitamin K.
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Imatinib-induced thyroiditis in Philadelphia chromosome-positive chronic myeloid leukemia p. 458
Surjit Singh, Pramod Kumar Sharma
DOI:10.4103/0253-7613.186214  
Here, we present a case of chronic myeloid leukemia for which imatinib therapy was initated. Triiodothyronine (T3), thyroxine (T4), and thyroid-stimulating hormone was normal, and thyroid microsomal autoantibodies (TMA) were positive and patient was diagnosed as thyroiditis treated with corticosteroids for 1½ months which lead to resolution.
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Phenytoin-induced encephalopathy in a child p. 460
Sumit Mehndiratta
DOI:10.4103/0253-7613.186209  
Phenytoin is a commonly used antiepileptic medication in the pediatric age group, but it has a narrow therapeutic range. Various adverse effects have been reported commonly. We report a relatively rare case of encephalopathy in a child from overdose of injectable phenytoin due to ignorance of the previous treatment. Scrutiny of medical records and history is of utmost importance while administering such medications.
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Torsades de pointes induced by concomitant use of chlorpheniramine and propranolol: An unusual presentation with no QT prolongation p. 462
Altug Osken, Nizamettin Selcuk Yelgec, Regayip Zehir, Tugba Kemaloglu Oz, Selcuk Yaylaci, Ramazan Akdemir, Huseyin Gunduz
DOI:10.4103/0253-7613.186193  
Drug-induced torsades de pointes (TdP) is a rare but potentially fatal adverse effect of commonly prescribed medications including cardiac and noncardiac drugs. Importantly, many drugs have been reported to cause the characteristic Brugada syndrome-linked electrocardiography (ECG) abnormalities and/or (fatal) ventricular tachyarrhythmias. Chlorpheniramine and propranolol have the arrhythmogenic effects reported previously. A review of literature revealed a large number of case reports of chlorpheniramine or propranolol use resulting in QTc prolongation, TdP, or both. However, we wish to report the case of a patient who was treated with a combination of chlorpheniramine and propranolol, whose ECG showed no QT prolongation but who suffered from cardiac arrest due to TdP.
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LETTERS TO THE EDITOR Top

Bedaquiline versus placebo for management of multidrug-resistant tuberculosis: A systematic review p. 466
Surjit Singh, Pramod Kumar Sharma, Rimplejeet Kaur
DOI:10.4103/0253-7613.186196  
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Author Reply p. 467
Jaykaran Charan, Tea Reljic, Ambuj Kumar
DOI:10.4103/0253-7613.186197  
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Irrational fixed dose combinations of psychotropic drugs in India: Cause of concern p. 468
Madhusudan Singh Solanki, Girish Banwari
DOI:10.4103/0253-7613.186192  
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Psychobiotics: A paradigm shift in psychopharmacology p. 469
Suravi Patra
DOI:10.4103/0253-7613.186194  
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OBITUARY Top

Dr. H. Venkata Krishna Bhatt, PhD FAMS FZS (August 12, 1942, to October 28, 2014) p. 471
P.S.R.K. Haranath
DOI:10.4103/0253-7613.186217  
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