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   2010| October-December  | Volume 2 | Issue 4  
    Online since October 28, 2010

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Introduction to metallic nanoparticles
Vicky V Mody, Rodney Siwale, Ajay Singh, Hardik R Mody
October-December 2010, 2(4):282-289
DOI:10.4103/0975-7406.72127  PMID:21180459
Metallic nanoparticles have fascinated scientist for over a century and are now heavily utilized in biomedical sciences and engineering. They are a focus of interest because of their huge potential in nanotechnology. Today these materials can be synthesized and modified with various chemical functional groups which allow them to be conjugated with antibodies, ligands, and drugs of interest and thus opening a wide range of potential applications in biotechnology, magnetic separation, and preconcentration of target analytes, targeted drug delivery, and vehicles for gene and drug delivery and more importantly diagnostic imaging. Moreover, various imaging modalities have been developed over the period of time such as MRI, CT, PET, ultrasound, SERS, and optical imaging as an aid to image various disease states. These imaging modalities differ in both techniques and instrumentation and more importantly require a contrast agent with unique physiochemical properties. This led to the invention of various nanoparticulated contrast agent such as magnetic nanoparticles (Fe 3 O 4 ), gold, and silver nanoparticles for their application in these imaging modalities. In addition, to use various imaging techniques in tandem newer multifunctional nanoshells and nanocages have been developed. Thus in this review article, we aim to provide an introduction to magnetic nanoparticles (Fe 3 O 4 ), gold nanoparticles, nanoshells and nanocages, and silver nanoparticles followed by their synthesis, physiochemical properties, and citing some recent applications in the diagnostic imaging and therapy of cancer.
  38,248 2,007 271
Orphan drug: Development trends and strategies
Aarti Sharma, Abraham Jacob, Manas Tandon, Dushyant Kumar
October-December 2010, 2(4):290-299
DOI:10.4103/0975-7406.72128  PMID:21180460
The growth of pharma industries has slowed in recent years because of various reasons such as patent expiries, generic competition, drying pipelines, and increasingly stringent regulatory guidelines. Many blockbuster drugs will loose their exclusivity in next 5 years. Therefore, the current economic situation plus the huge generic competition shifted the focus of pharmaceutical companies from the essential medicines to the new business model - niche busters, also called orphan drugs. Orphan drugs may help pharma companies to reduce the impact of revenue loss caused by patent expiries of blockbuster drugs. The new business model of orphan drugs could offer an integrated healthcare solution that enables pharma companies to develop newer areas of therapeutics, diagnosis, treatment, monitoring, and patient support. Incentives for drug development provided by governments, as well as support from the FDA and EU Commission in special protocols, are a further boost for the companies developing orphan drugs. Although there may still be challenges ahead for the pharmaceutical industry, orphan drugs seem to offer the key to recovery and stability within the market. In our study, we have compared the policies and orphan drug incentives worldwide alongwith the challenges faced by the pharmaceutical companies. Recent developments are seen in orphan drug approval, the various drugs in orphan drug pipeline, and the future prospectives for orphan drugs and diseases.
  16,949 3 44
Emerging fungal infections among children: A review on its clinical manifestations, diagnosis, and prevention
Akansha Jain, Shubham Jain, Swati Rawat
October-December 2010, 2(4):314-320
DOI:10.4103/0975-7406.72131  PMID:21180463
The incidence of fungal infections is increasing at an alarming rate, presenting an enormous challenge to healthcare professionals. This increase is directly related to the growing population of immunocompromised individuals especially children resulting from changes in medical practice such as the use of intensive chemotherapy and immunosuppressive drugs. Although healthy children have strong natural immunity against fungal infections, then also fungal infection among children are increasing very fast. Virtually not all fungi are pathogenic and their infection is opportunistic. Fungi can occur in the form of yeast, mould, and dimorph. In children fungi can cause superficial infection, i.e., on skin, nails, and hair like oral thrush, candida diaper rash, tinea infections, etc., are various types of superficial fungal infections, subcutaneous fungal infection in tissues under the skin and lastly it causes systemic infection in deeper tissues. Most superficial and subcutaneous fungal infections are easily diagnosed and readily amenable to treatment. Opportunistic fungal infections are those that cause diseases exclusively in immunocompromised individuals, e.g., aspergillosis, zygomycosis, etc. Systemic infections can be life-threatening and are associated with high morbidity and mortality. Because diagnosis is difficult and the causative agent is often confirmed only at autopsy, the exact incidence of systemic infections is difficult to determine. The most frequently encountered pathogens are Candida albicans and Aspergillus spp. But other fungi such as non-albicans Candida spp. are increasingly important.
  15,618 407 39
Fast dissolving strips: A novel approach for the delivery of verapamil
S Kunte, P Tandale
October-December 2010, 2(4):325-328
DOI:10.4103/0975-7406.72133  PMID:21180465
Objective: Fast dissolving drug delivery system offers a solution for those patients having difficulty in swallowing tablets/capsules etc. Verapamil is a calcium channel blocker used as an antianginal, antiarrhythmic, and antihypertensive agent with extensive first pass metabolism which results in less bioavailability. This work investigated the possibility of developing verapamil fast dissolving strips allowing fast, reproducible drug dissolution in the oral cavity; thus bypassing first pass metabolism. Materials and methods: The fast dissolving strips were prepared by solvent casting technique with the help of HPMC E6 and maltodextrin. The strips were evaluated for drug content uniformity, film thickness, folding endurance, in vitro disintegration time, in vitro dissolution studies, surface pH study, and palatability study. Results: Official criteria for evaluation parameters were fulfilled by all formulations. Disintegration time showed by formulations was found to be in range of 20.4-28.6 sec. Based on the evaluation parameters, the formulation containing 2% HPMC E6 and 3.5% maltodextrin showed optimum performance against other formulations. Conclusion: It was concluded that the fast dissolving strips of verapamil can be made by solvent casting technique with enhanced dissolution rate, taste masking, and hence better patient compliance and effective therapy
  11,891 773 39
Sensitive and selective method for the analysis of menthol from pharmaceutical products by RP-HPLC with refractive index detector
KA Shaikh, Sachin D Patil
October-December 2010, 2(4):360-364
DOI:10.4103/0975-7406.72141  PMID:21180473
Objective : Liquid chromatography with refractive index (RI) detection has been found to be very useful for the determination of menthol from pharmaceutical products. A simple and rapid HPLC method has been developed for this purpose compared to conventional GC methods, requiring no special sample pretreatment for the determination of menthol from pharmaceutical products. Materials and Methods : A chromatographic separation was achieved on a Inertsil ODS 3V (4.6mm×250mm, 5μm) column using water : methanol (30:70 v/v) as a mobile phase, at a flow rate of 1.0 ml/min. Results : Method was validated as per ICH guidelines for various parameters such as precision, linearity, accuracy, solution stability, robustness, limit of detection and quantification. Results were found to be within acceptable limits. Conclusion : The method has been successfully applied for the quantification of menthol from syrup formulations. The developed method can be conveniently used by the quality control department to determine assay of menthol from pharmaceutical preparations.
  9,590 290 10
Transdermal drug delivery of labetalol hydrochloride: Feasibility and effect of penetration enhancers
Saqib Zafar, Asgar Ali, Mohammed Aqil, Abdul Ahad
October-December 2010, 2(4):321-324
DOI:10.4103/0975-7406.72132  PMID:21180464
Objectives : The objective of this study is to investigate the feasibility of transdermal drug delivery of Labetalol Hydrochloride (LHCl) and to study the effect of different penetration enhancers on the skin permeability of LHCl. Methods : The permeability experiments were conducted using a horizontal glass diffusion cell with a diffusional area of 2.37 cm­2 on albino rat skin. The effect of various penetration enhancers namely turpentine oil, dimethyl formamide (DMF), menthol, dimethyl sulfoxide, pine oil, and 2-pyrollidone, and the effect of the concentration of drug and enhancer in the donor phase on the skin permeability of LHCl was studied. Results : The apparent partition coefficient of the drug was found to be 6.95, suggesting it to be a lipophilic drug. The preliminary skin permeation studies revealed that the permeation of LHCL through albino rat skin was moderate (Kp = 6.490 Χ 10 -2 cm hr -1 ) from isotonic phosphate buffer of pH 7.4. An appreciable increase in the LHCl permeability coefficient was observed on using a co-solvent (ethanol 95%) with the penetration enhancers in the donor phase. DMSO (10% v/v) was found to be the most effective enhancer for Labetalol hydrochloride (Enhancement Factor = 1.165). An increase in the concentration of drug and enhancer in the donor cell accentuated the permeability coefficient of LHCl. Conclusions : It was concluded that LHCl could be delivered via the dermal route with the use of 10% DMSO as the penetration enhancer.
  9,533 237 7
Food and drug administration's critical path initiative and innovations in drug development paradigm: Challenges, progress, and controversies
Rajiv Mahajan, Kapil Gupta
October-December 2010, 2(4):307-313
DOI:10.4103/0975-7406.72130  PMID:21180462
During the last decade, despite increased investment in drug research and development related activity, stagnation in new drug discovery has been documented. Despite a 70% increase in investment in research and development-related activities, a 40% fall in launch of new chemical entities was seen during 1994-2004. A steep rise in the attrition rate of drug development has complicated the matter. Rising cost and increased attrition rates proved major barriers to investment in higher risk drugs or in therapies for uncommon diseases or diseases that predominantly afflict the poor. This prompted Food and Drug Administration (FDA) to highlight this problem in a 2004 white paper classified as "Critical Path Initiative" (CPI) and to initiate steps to target stagnation and rise in attrition rates. Many new drug development projects have started worldwide taking cue from CPI; adopting microdosing, adaptive designs and taking advantage of newly developed biomarkers under the CPI. This review discusses the various strategies adopted under CPI to decrease attrition rate and stagnation of new drug development, and the challenges and controversies associated with CPI.
  6,015 298 18
Contribution of CNS cells in NeuroAIDS
Ashish Swarup Verma, Udai Pratap Singh, Premendra Dhar Dwivedi, Anchal Singh
October-December 2010, 2(4):300-306
DOI:10.4103/0975-7406.72129  PMID:21180461
NeuroAIDS is becoming a major health problem among AIDS patients and long-term HIV survivors. As per 2009 estimates of UNAIDS report, more than 34 million people have been infected with HIV out of which ≥ 50% show signs and symptoms of neuropsychiatric disorders. These disorders affect central nervous system (CNS) and peripheral nervous systems (PNS). CNS is one of the most protected organ systems in body which is protected by blood-brain barrier (BBB). Not only this, most of the cells of CNS are negative for receptors and co-receptors for HIV infections. Neurons have been found to be completely nonpermissive for HIV infection.These facts suggest that neurotoxicity could be an indirect mechanism responsible for neuropsychiatric complications. In this review, we will discuss the importance of different cell types of CNS and their contribution toward neurotoxicity.
  5,645 121 8
Spectrophotometric methods for determination of zolpidem tartrate in tablet formulation
Rajiv Chomwal, Amit Kumar, Anju Goyal
October-December 2010, 2(4):365-368
DOI:10.4103/0975-7406.72142  PMID:21180474
Aim : The study aims to develop simple, sensitive, rapid, accurate and precise spectrophotometric method for estimation of Zolpidem tartrate in tablet dosage forms. Materials and Methods : For method I, II, III and IV in a series of 10 ml volumetric flask, aliquots of standard drug solution (100 μg/ml) in 0.1N HCl were transferred and diluted with same so as to give several dilutions in concentration range of 5-30 μg/ml, 5-30 μg/ml, 10-50 μg/ml and 5-40 μg/ml respectively of zolpidem tartrate. To 5 ml of each dilution taken in a separating funnel, (5 ml of bromo phenol red, bromo cresol purple, bromo cresol green and bromo phenol blue for method I, II, III and IV respectively) reagent and 5 ml of chloroform was added. Reaction mixture was shaken gently for 5 min and allowed to stand so as to separate aqueous and chloroform layer. Absorbance maxima measured at 407 nm, 417 nm, 412 nm and 415 nm for method I, II, III and IV respectively. Results : The recovery studies were found close to 100 % that indicates accuracy and precision of the proposed methods. The statistical analysis was carried out and results of which were found satisfactory. Standard deviation values were found low that indicated reproducibility of the proposed methods. Conclusion : Based on results the developed methods could be used for routine estimation of zolpidem tartrate from tablet formulations.
  5,024 182 2
Spectrophotometric method for simultaneous estimation of atenolol in combination with losartan potassium and hydrochlorothiazide in bulk and tablet formulation
Sanjay Bari, Shital Sathe, Pritam Jain, Sanjay Surana
October-December 2010, 2(4):372-375
DOI:10.4103/0975-7406.72144  PMID:21180476
Aim : To develop a simple, accurate, rapid and precise UV spectrophotometric method for the estimation of atenolol in combination with losartan potassium and hydrochlorothiazide. Materials and Methods : The method employs formation and solving simultaneous equation using 251.60 nm and 224.20 nm for losartan potassium and atenolol, 224.20 and 271.60 for atenolol and hydrochlorothiazide as two analytical wavelengths, using methanol water as a solvent. Results and Conclusion : The linearity was observed in the concentration range of 5-30 μg/ml (r=0.9991) for losratan pottassium, 2 - 12 μg/ml (r = 0.9995) for atenolol and 2 - 14 μg/ml (r = 0.9993) for hydrochlorothiazide. The results of the method were validated statistically and by recovery studies.
  4,848 242 4
Effect of Erythrina variegata seed extract on hyperlipidemia elicited by high-fat diet in wistar rats
G Balamurugan, A Shantha
October-December 2010, 2(4):350-355
Objective : To investigate the effect of the methanolic extract of Erythrina variegata (Linn.) var Orientalis (Fabaceae) seeds (MEEV) in reducing the cholesterol levels and as well as antioxidant in experimentally induced hyperlipidemic rats. Materials and Methods : Doses of 200 and 400 mg/kg of the extract were evaluated for its effect on lipid profile, HMG-CoA reductase, and on antioxidant enzymes in high-fat diet (HFD) induced hyperlipidemia. Results and Conclusion : The elevated levels of total cholesterol, triglycerides, low-density lipoprotein, and very low density lipoprotein due to HFD was reduced by concurrent treatment with MEEV (200 and 400 mg/kg) significantly (P<0.001). A significant reduction (P<0.001) in high-density lipoprotein was noticed in HFD fed groups; however, a nonsignificant increment was produced by the administration of MEEV (200 and 400 mg/kg). The HMG-CoA reductase activity was increased in HFD fed animals significantly (P<0.001) and was reduced by MEEV 400 mg/kg significantly (P<0.001). There was a noticed increase in the body weight and mesenteric fat pad weight in HFD fed group (P<0.001), which was reduced by the administration of MEEV (200 and 400 mg/kg). The antioxidant enzymes such as superoxide dismutase and catalase were reduced significantly in the HFD fed group, whose levels were increased significantly (P<0.001) by the administration of MEEV (200 and 400 mg/kg). Lipid peroxidation was increased in HFD fed animals, which was reduced significantly (P<0.001) by the treatment with MEEV (200 and 400 mg/kg).
  4,542 223 -
A pharmacovigilance study on patients of bronchial asthma in a teaching hospital
AN Jamali, M Aqil, MS Alam, KK Pillai, P Kapur
October-December 2010, 2(4):333-336
DOI:10.4103/0975-7406.72135  PMID:21180467
Objective : The present study was conducted to monitor adverse drug reactions in patients of bronchial asthma in outpatient department and inpatient department of a university teaching hospital in South Delhi. Materials and Methods : About 200 patients irrespective of age and sex with established asthma were interviewed during the time period of January 2006 to April 2006 using structured questionnaire. Naranjo's adverse drug reaction probability scale was used to assess the adverse drug reactions. Results : A total of 15 adverse drug reactions were reported in 13 out of 200 asthmatic patients. Among the 13 patients reported with adverse drug reactions, 5 (38.5%) were male and 8 (61.5%) patients were female. Maximum percentage of ADRs (2 in 15 prescriptions, 13.3%) observed with montelukast, followed by beclomethasone (1 in 12 prescriptions, 8.3%), salbutamol (6 in 109 prescriptions, 5.5%), and ipratropium (3 in 63 prescriptions, 4.8%). Conclusions : Montelukast was found to be associated with greater percentage of adverse drug reactions as compared to other antiasthamatics. The above findings are constrained by a small sample size and need to be corroborated by conducting long-term studies using a larger sample size.
  4,303 196 2
Evaluation of immunomodulatory potential of ethanolic extract of Roscoea procera rhizomes in mice
Mahesh S Sahu, Prashant Y Mali, Shekhar B Waikar, Vinod D Rangari
October-December 2010, 2(4):346-349
DOI:10.4103/0975-7406.72138  PMID:21180470
Purpose : The aim of present study was to evaluate immunomodulatory potential of ethanolic extract of Roscoea procera (Zingiberaceae) rhizomes by using delayed-type hypersensitivity (DTH) and carbon clearance method in comparison to standard established immunosuppressant drug, cyclophosphamide (30 mg/kg, i.p.) in mice. Material and Methods : The extract was comprised to acute toxicity (OECD-423 guideline), DTH and carbon clearance method for their immunomodulatory potential. Ethanolic extract of Roscoea procera rhizomes administered orally at doses 300 mg/kg and 600 mg/kg, p.o. to mice. Result and Conclusion : Result of our study revealed that, the foot pat thickness of ethanolic extract group (P<0.05) significantly enhanced the production of circulating antibody titre in response to Sheep red blood cells (SRBC) and phagocytic functions of mononuclear macrophages and non-specific immunity. Result were also supported by serological and haematological tests data. Hence, the present investigation reveals that, ethanolic extract of Roscoea procera rhizomes possesses immunostimulant properties. Further studies to identify the active moieties and elucidation of the mechanism of action are recommended.
  4,079 248 9
Assessment of asthma and chronic obstructive pulmonary disorder in relation to reversibility, IgE, eosinophil, and neutrophil count in a University Teaching Hospital in South Delhi, India
Virender P. Singh Rathod, Prem Kapoor, KK Pillai, Razia Khanam
October-December 2010, 2(4):337-340
DOI:10.4103/0975-7406.72136  PMID:21180468
Objectives : The physiological and clinical similarities between asthma and chronic obstructive pulmonary disorder (COPD) make their differentiation difficult. In the present study, we compared reversibility to bronchodilator, immunoglobulin E (IgE), blood eosinophil and neutrophil levels among asthma and COPD patients to differentiate these diseases. Materials and Methods : The study was carried on 20 asthmatics and 29 patients of COPD that reported to the outpatient and inpatient department in University Teaching Hospital, Jamia Hamdard, New Delhi, India. The parameters evaluated included pulmonary function (FEV 1 , FVC, and FEV 1 /FVC), IgE levels, and eosinophil and neutrophil count. Results : It was observed that reversibility was significantly higher in asthmatic patients, while irreversibility predominates in COPD patients. There was no significant difference in pre- and post-FEV 1 and pre- and post-FVC and in their percentage predicted. However the percentage change in FEV 1 significantly varies in asthma and COPD patients. No significant changes in neutrophil and eosinophil levels were observed in these patients. The serum IgE levels were found significantly higher in asthmatic patients. Conclusions : We conclude that reversibility in FEV 1 levels or percentage change in FEV 1 and serum IgE levels are promising lab parameter to distinguish these two conditions. However, further research is required to fully understand the role of neutrophil and eosinophil in the onset and development of asthma and COPD.
  4,193 125 3
Hepatoprotective potential of Decalepis hamiltonii (Wight and Arn) against carbon tetrachloride-induced hepatic damage in rats
R Harish, T Shivanandappa
October-December 2010, 2(4):341-345
DOI:10.4103/0975-7406.72137  PMID:21180469
Hepatoprotective activity of the roots of Decalepis hamiltonii (Wight and Arn) was studied using carbon tetrachloride (CCl 4 ) induced liver injury model in albino rats. The hepatotoxicity produced by acute CCl 4 administration was found to be inhibited by pretreating the rats with crude methanolic extract of the roots of D. hamiltonii (Dh) prior to CCl 4 induction. Hepatotoxic inhibition was measured with the decreased levels of hepatic serum marker enzymes (glutamate-pyruvate transaminase (GPT), glutamate oxaloacetate transaminase (GOT), alkaline phosphatase (ALP), and lactate dehydrogenase (LDH) and lipid peroxide formation. Imbalance level of glutathione (GSH) and antioxidant enzymes such as catalase, glutathione peroxidase, and glutathione reductase were normalized in rats pretreated with Dh extract followed by CCl 4 administration. Pathological changes of hepatic lesions caused by CCl 4 were also improved by pretreatment with the Dh root extract. The results of this study indicate that roots of D. hamiltonii could afford a significant protective action in the alleviation of CCl 4 -induced hepatic damage in rats.
  3,680 200 9
Journal of Pharmacy and Bioallied Sciences
Mohd. Aqil
October-December 2010, 2(4):281-281
DOI:10.4103/0975-7406.72126  PMID:21180458
  3,332 156 -
Neuropharmacological safety evaluation of jigrine: A polyherbal hepatoprotective formulation
AK Najmi, KK Pillai, SN Pal, M Akhtar, M Mujeeb, A Aftab
October-December 2010, 2(4):329-332
DOI:10.4103/0975-7406.72134  PMID:21180466
Objective : Jigrine is a herbal hepatoprotective formulation containing aqueous extracts of 14 medicinal plants. Present study was designed to evaluate per se neuropharmacological effects of jigrine in mice. Materials and Methods : Jigrine was evaluated in a number of pharmacological test paradigms, viz. open field arena, actophotometer, hole board, rotarod, traction test, grip strength test, spontaneous alternation behavior, passive avoidance task, and phenobarbital sleeping time. Results and Conclusions : Jigrine pretreatment (1 and 2 ml/kg, p.o.) did not produce any significant effect as compared to normal saline treated animals and was found to be free from any acute undesirable central effects at these two dose levels.
  3,298 132 2
Monitorization of drug content in furosemide and lorazepam tablets stored in multidose pill boxes
Jessica Martins, Magda Oliveira, Maria Tapiço, Tânia Nascimento, Ana Grenha, Luis Braz
October-December 2010, 2(4):356-359
DOI:10.4103/0975-7406.72140  PMID:21180472
Background : Therapeutic nonadherence is a major health problem, particularly when therapeutic regimens are complex and long-lasting. Therefore, tools such as multidose pill boxes have been designed to provide the means for higher therapeutic compliance. However, no studies are available reporting on their capacity to keep the drug content of the stored tablets unaltered. Objective : This work aimed at monitoring the drug content of tablets stored in multidose boxes for a period of two weeks. Materials and Methods : Furosemide and lorazepam were selected as model drugs, given their frequent chronic use, which is coherent with the profile of medicines susceptible of storage in the referred boxes. Variations of the tablets drug content were assessed as a function of temperature (25°C and 40°C) and the presence of blister. Results and Discussion : The obtained results allowed concluding that concerning temperature, only lorazepam tablets registered drug content alterations and only when stored at 40°C. On the other side, it was concluded that the absence of blister does not compromise the drug content of the studied tablets. Conclusion : In the specific conditions of this study, the storage of these medicines in multidose boxes is considered reliable and adequate.
  3,238 73 1
Fluorescent labeling of human albumin using the new aromatic dialdehyde labels and the study of innerfilter effect
Muhammad Aminuddin, Sofia Ahmed, Muhammd Ali Sheraz, Iqbal Ahmad, Karamat Mahmood, JN Miller
October-December 2010, 2(4):369-371
DOI:10.4103/0975-7406.72143  PMID:21180475
The labels naphthalene-2,3-dicarboxaldehyde (NDA), 1-phenylnaphthalene-2,3-dialdehyde (fNDA), and anthracene-2,3-dialdehyde (ADA) have been used as fluorigenic reagents. They formed fluorescent derivatives with proteins. The derivatives formed are in fact isoindoles. The fluorescence decay of the labels-antibody was found to extend over a period of 4, 8, and 10 h for ΦNDA, ADA, and NDA-derivative, respectively. Protein formed is comparatively less stable as compared to simple amino acids. In relation to innerfilter effect, the addition of cytochrome C, myoglobin, and ATP as absorbers to label-human albumin fluorophores appeared to have quenched the fluorescence. In the case of using NDA as label, the fluorescence was quenched roughly 70%, 24%, and 58% for addition of cytochrome C, myoglobin, and ATP, respectively. The labels used were found to give rapid, reproducible, and reliable results.
  2,753 73 -
Bacterial virulence traits: A potential area of study for drug development
Roopesh Jain, Susmit Kosta, Archana Tiwari
October-December 2010, 2(4):376-376
DOI:10.4103/0975-7406.72145  PMID:21180477
  2,366 94 -
Periodontitis leads to VAP in ICU patients: A dental note
Rajiv Saini, Santosh Saini, Sugandha Sharma
October-December 2010, 2(4):377-377
DOI:10.4103/0975-7406.72146  PMID:21180478
  2,398 59 2