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   2012| April-June  | Volume 4 | Issue 2  
    Online since April 10, 2012

 
 
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ORIGINAL ARTICLES
Moringa oleifera Lam. (Moringaceae) grown in Nigeria: In vitro antisickling activity on deoxygenated erythrocyte cells
Olufunmilayo E Adejumo, Adelodun L Kolapo, Akintomiwa O Folarin
April-June 2012, 4(2):118-122
DOI:10.4103/0975-7406.94812  
Context: Traditional medicine, which is more available and affordable for the poor uses medicinal plants for the treatment and management of various ailments, including the sickle cell disease (SCD). About 24 million Nigerians are carriers of this sickled cell gene, while approximately 2.4 million are SCD patients. Moringa oleifera Lam. (Moringaceae) possesses high nutritional value and has been used in folklore medicine to treat various ailments related to pain and inflammation. Chemical, pharmacological and pharmacognostical applications of Moringa oleifera have been reported. Objective: This study investigated the antisickling potential of polar and non-polar extracts of the seed, flower and leaf of Moringa oleifera for the first time. Materials and Methods: Using crude methanol extract, aqueous extract, ethyl acetate and butanol, the in vitro antisickling activities of Moringa oleifera fractions, were evaluated using erythrocyte cells deoxygenated with 2% sodium metabisulphite. p-Hydroxybenzoic acid and normal saline were employed as positive and negative controls. Results: Phytochemical screening revealed the presence of saponins, free anthraquinones, and alkaloids. Extracts of the seed and flower demonstrated a higher (P<0.05) antisickling activity in comparison to the leaf extract. The leaf extract, as well as those of the seed and flower, equally demonstrated a (P<0.05) reversal of sickled erythrocytes. Discussions and Conclusions: These findings suggest that Moringa oleifera may play a role in the management of SCD, by incorporation of its fractions into recipes. More extensive biological evaluations and further studies will be necessary for the chemical characterization of the antisickling principles.
  8,546 188 10
Preparation and in vitro/in vivo characterization of curcumin microspheres intended to treat colon cancer
M Madhavi, K Madhavi, AV Jithan
April-June 2012, 4(2):164-171
DOI:10.4103/0975-7406.94825  
Objective: The objective of the present investigation was to prepare colon targeted curcumin microspheres using Eudragit S100 and evaluate the same for in vitro/in vivo properties. Materials and Methods: A "O/O solvent evaporation" technique was used in the preparation of microspheres. The influence of various process variables including stirring speed, drug:polymer ratio and percentage of emulsifier on the fabrication were investigated and the formulation was optimized. Prepared microspheres were evaluated for in vitro and in vivo properties. Surface morphology, particle size, percentage drug entrapment, percentage yield, drug polymer interaction, in vitro drug release in simulated gastrointestinal transit conditions and stability were the in vitro parameters investigated. Using an optimized formulation, drug release into the systemic circulation and organ distribution were investigated as in vivo parameters. In vivo parameters were estimated in male albino rats. Results: Curcumin microspheres of Eudragit S100 were successfully prepared using o/o solvent evaporation method. Microspheres prepared using 1:2 drug:polymer ratio, with a stirring speed of 1000 rpm, and using 1.0% w/v concentration of emulsifying agent was selected as an optimized formulation. The release studies with optimized formulation demonstrated that aqueous solubility of curcumin was enhanced by 8 times with the formulation. FTIR studies demonstrated no change in drug characteristics upon microsphere fabrication. The enhancement in solubility is thus due to the increase in the surface area of the drug substance and not due to a change of drug to a different physical state. This was further confirmed by scanning electron microsphere pictures. Drug release followed Korsmeyer and Peppas release model. Accelerated stability studies indicated that the drug is stable in the formulation for a period of atleast 14 weeks at room temperature. In vivo studies demonstrated a sustained drug release into the systemic circulation after oral administration of the formulation. Further, colon target was affectively achieved using the optimized formulation. Eudragit microspheres delivered most of their drug load (79.0%) to the colon, whereas with plain drug suspension only 28.0% of the total dose reached the target site. Conclusion: This study successfully developed curcumin microspheres that can be used effectively in the treatment of the colon cancer.
  5,444 393 11
Economic evaluation of end stage renal disease patients undergoing hemodialysis
A Suja, R Anju, V Anju, J Neethu, P Peeyush, R Saraswathy
April-June 2012, 4(2):107-111
DOI:10.4103/0975-7406.94810  
Background: In India the incidence of end stage renal disease (ESRD) is increasing day by day and the option for the treatment of ESRD is dialysis or transplantation. In the present scenario, due to the cost of treatment normal people can afford only hemodialysis rather than transplantation. Since the cost of hemodialysis differs across the country, research is needed to evaluate its exact cost. Aim: This study is to analyze the healthcare cost of hemodialysis in a private hospital of South India. Materials and Methods: This is a prospective, observational study carried out in a tertiary care hospital. Patients who are undergoing routine hemodialysis in this hospital were selected for the study. Patient data as well as cost details were collected for a period of six months. Thirty patients were selected for the study and a total of 2160 dialysis sessions were studied. Patient perspective was taken for the analysis of cost. Both direct and indirect costs were analyzed. This includes cost of dialysis, investigations, erythropoietin, food, transportation, lost wages etc. Socioeconomic status of the patient was also studied. Result: The total cost per session was found to be around Rs. 4500. Fifty six percent contributes direct medical cost whereas 20% contributes direct non medical cost. Twenty four percent cost was due to indirect costs. Since the patients are paying from their own pocket, only the upper or upper middle class patient can undergo hemodialysis regularly. Conclusion: These findings are important to find out the impact of cost of hemodialysis on patients suffering from ESRD. Further studies related to costs and outcome, otherwise known as pharmacoeconomic studies, are needed to analyze the pros and cons of renal replacement therapy and to improve the quality of life of ESRD patients. Thus pharmacoeconomical studies are needed to realize that government has to take initiative to provide insurance or reimbursement for the common people.
  5,250 293 8
Development of span 80-tween 80 based fluid-filled organogels as a matrix for drug delivery
Charulata Bhattacharya, Nikhil Kumar, Sai S Sagiri, Kunal Pal, Sirsendu S Ray
April-June 2012, 4(2):155-163
DOI:10.4103/0975-7406.94822  
Background: Organogels are defined as 3-dimensional networked structures which immobilize apolar solvents within them. These gelled formulations are gaining importance because of their ease of preparation and inherent stability with improved shelf life as compared to the ointments. Aim: Development of span 80-tween 80 mixture based organogels for the first time by fluid-filled fiber mechanism. Materials and Methods: Span 80 and tween 80 were used as surfactant and co-surfactant, respectively. The surfactant mixtures were dissolved in oil followed by the addition of water which led to the formation of organogels at specific compositions. The formulations were analyzed by microscopy, X-ray diffraction (XRD), time-dependent stability test and accelerated thermal stability test by thermocycling method. Ciprofloxacin, a fourth-generation fluoroquinolone, was incorporated within the organogels. The antimicrobial activity of the drug loaded organogels and in vitro drug release from the gels was also determined. Results and Conclusions: Microscopic results indicated that the gels contained clusters of water-filled spherical structures. XRD study indicated the amorphous nature of the organogels. The release of the drug was found to be diffusion controlled and showed marked antimicrobial property. In short, the prepared organogels were found to be stable enough to be used as pharmaceutical formulation.
  4,255 201 16
Simultaneous HPTLC determination of strychnine and brucine in strychnos nux-vomica seed
Abid Kamal, YT Kamal, Sayeed Ahmad, FJ Ahmad, Kishwar Saleem
April-June 2012, 4(2):134-139
DOI:10.4103/0975-7406.94814  
Objective: A simple, sensitive, and specific thin layer chromatography (TLC) densitometry method has been developed for the simultaneous quantification of strychnine and brucine in the seeds of Strychnos nux-vomica. Materials and Methods: The method involved simultaneous estimation of strychnine and brucine after resolving it by high performance TLC (HPTLC) on silica gel plate with chloroform-methanol-formic acid (8.5:1.5:0.4 v/v/v) as the mobile phase. Results: The method was validated as per the ICH guidelines for precision (interday, intraday, intersystem), robustness, accuracy, limit of detection, and limit of quantitation. The relationship between the concentration of standard solutions and the peak response was linear within the concentration range of 50-1000 ng/spot for strychnine and 100-1000 ng/spot for brucine. The method precision was found to be 0.58-2.47 (% relative standard deviation [RSD]) and 0.36-2.22 (% RSD) for strychnine and brucine, respectively. Accuracy of the method was checked by recovery studies conducted at three different concentration levels and the average percentage recovery was found to be 100.75% for strychnine and 100.52% for brucine, respectively. Conclusions: The HPTLC method for the simultaneous quantification of strychnine and brucine was found to be simple, precise, specific, sensitive, and accurate and can be used for routine analysis and quality control of raw material of S. nux-vomica and several unani and ayurvedic formulations containing this as an ingredient.
  4,192 175 6
New evidence on α-synuclein and Tau binding to conformation and sequence specific GC* rich DNA: Relevance to neurological disorders
P Vasudevaraju, Erika Guerrero, Muralidhar L Hegde, TB Collen, Gabrielle B Britton, KS Rao
April-June 2012, 4(2):112-117
DOI:10.4103/0975-7406.94811  
Background: Deoxyribonucleic acid (DNA) topology plays a critical role in maintaining the integrity of the genome and cellular functions. Although changes in DNA conformation and structural dynamics in the brain have been associated with various neurological disorders, its precise role in the pathogenesis is still unclear. Previous studies from our laboratory have shown that there is a conformational change in the genomic DNA of Parkinson's disease (PD) (B to altered B-DNA) and Alzheimer's disease brain (B to Z-DNA). However, there is limited information on the mechanism on DNA dynamics changes in brain. Objective: In the present study, we have investigated the DNA conformation and sequence specific binding ability of α-Synuclein and Tau with reference to B-DNA and Z-DNA using oligonucleotide (CGCGCGCG) 2 as a novel model DNA system. This sequence is predominantly present in the promoter region of the genes of biological relevance. Materials and Methods: Natively, (CGCGCGCG) 2 sequence exists in B-DNA conformation, but in the presence of high sodium concentration (4 M NaCl), the oligo converts into Z-DNA form. We used circular dichroism, melting temperature and fluorescence studies to understand protein-DNA interactions. Results: CD studies indicated that both α-Synuclein and Tau bind to B-DNA conformation of (CGCGCGCG) 2 and induce altered B-form. Further, these proteins increased the melting temperature and decreased the number of EtBr molecules bound per base pair of DNA in B-form indicating that DNA stability is favored to alter B-DNA conformation, which could be an intermediate form favoring Z-DNA conformation. Moreover, both α-Synuclein and Tau also bound to disease-linked Z-DNA conformation of (CGCGCGCG) 2 and further stabilized the Z-conformation. Conclusions: The present study provides vital mechanistic information on Synuclein and Tau binding to DNA in a conformation-specific manner causing conformational transition. Furthermore, both the proteins stabilize Z-DNA conformation. These have altered minor and major groove patterns and thus may have significant biological implications in relevance to gene expression pattern in neurodegeneration. We discuss the implications of α-Synuclein/Tau binding to DNA and stabilizing the altered conformations of DNA in neuronal cell dysfunction.
  3,779 85 10
Analgesic and cytotoxic activity of Acorus calamus L., Kigelia pinnata L., Mangifera indica L. and Tabernaemontana divaricata L.
Mohammad Ahad Ali Khan, Mohammad Torequl Islam
April-June 2012, 4(2):149-154
DOI:10.4103/0975-7406.94820  
Objectives: The aim of the study was to evaluate analgesic and cytotoxic activity of Acorus calamus L., Kigelia pinnata L., Mangifera indica L., Tabernaemontana divaricata L. extracts by using acetic acid-induced writhing method in mice and brine shrimp lethality assay. Materials and Methods: The ethanolic extracts of the plants were obtained by simple maceration method and were subjected to standardization by using pharmacognostical and phytochemical screening methods, which were followed by acetic acid writhing and brine shrimp lethality test methods. Dose selection was made on the basis of acute oral toxicity study (10-1000 mg/kg body weight). Results and Conclusion: In analgesic test, M. indica L. extract produced 28.16% and 22.02% writhing protection at the doses of 250 and 500 mg/kg body weight in mice, respectively. While the T. divaricata L. extract produced 22.02% and 33.93%, K. pinnata L. extract produced 11.55% and 47.29% and A. calamus L. extract produced 15.16% and 54.51% of writhing protection at the same doses. The percent mortality (mean ± SD) was found to be 58.7 ± 25.22, 56.25 ± 22.88, 52.50 ± 24.37, and 61.25 ± 26.66 with M. indica L., T. divaricata L., K. pinnata L., and A. calamus L., respectively. And the LC 50 and LC 90 values were found to be 100 and 300 μg/mL for M. indica L. and that were (200 and 350 μg/mL), (100 and 350 μg/mL) and (50 and 300 μg/mL) for T. divaricata L., K. pinnata L., and A. calamus L., respectively. Thus it can be concluded that bark of M. indica L., leaves of T. divaricata L., bark of K. pinnata L., and roots of A. calamus L. have significant analgesic and cytotoxic activity and can be preferred in the treatment of pain and tumor.
  3,313 162 8
Anticonvulsant and related neuropharmacological effects of the whole plant extract of Synedrella nodiflora (L.) Gaertn (Asteraceae)
Patrick Amoateng, Eric Woode, Samuel B Kombian
April-June 2012, 4(2):140-148
DOI:10.4103/0975-7406.94816  
Purpose: The plant Synedrella nodiflora (L) Gaertn is traditionally used by some Ghanaian communities to treat epilepsy. To determine if this use has merit, we studied the anticonvulsant and other neuropharmacological effects of a hydro-ethanolic extract of the whole plant using murine models. Materials and Methods: The anticonvulsant effect of the extract (10-1000 mg/kg) was tested on the pentylenetetrazole-, picrotoxin-, and pilocarpine-induced seizure models and PTZ-kindling in mice/rats. The effect of the extract was also tested on motor coordination using the rota-rod. Results: The results obtained revealed that the extract possesses anticonvulsant effects in all the experimental models of seizures tested as it significantly reduced the latencies to myoclonic jerks and seizures as well as seizure duration and the percentage severity. The extract was also found to cause motor incoordination at the higher dose of 1000 mg/kg. Conclusions: In summary, the hydro-ethanolic extract of the whole plant of S. nodiflora possesses anticonvulsant effects, possibly through an interaction with GABAergic transmission and antioxidant mechanisms and muscle relaxant effects. These findings thus provide scientific evidence in support of the traditional use of the plant in the management of epilepsy.
  3,040 134 3
Validated stability-indicating high-performance thin-layer chromatographic method for estimation of cefpodoxime proxetil in bulk and in pharmaceutical formulation according to International conference on harmonization guidelines
Pritam Jain, Amar Chaudhari, Anup Bang, Sanjay Surana
April-June 2012, 4(2):101-106
DOI:10.4103/0975-7406.94809  
Aim: A simple, selective, precise, and stability-indicating high-performance thin-layer chromatographic (HPTLC) method for analysis of cefpodoxime proxetil both in bulk and in pharmaceutical formulation has been developed and validated. Materials and Methods: The method employed HPTLC aluminum plates precoated with silica gel 60 RP-18 F 254 as the stationary phase. The solvent system consisted of toluene:methanol:chloroform (4:2:4 v/v). The system was found to give compact spot for cefpodoxime proxetil (R f value of 0.55 ± 0.02). Densitometric analysis of cefpodoxime proxetil was carried out in the absorbance mode at 289 nm. Results: The linear regression analysis data for the calibration plots showed good linear relationship, with r 2 = 0.998 ± 0.0015 with respect to peak area in the concentration range of 100-600 ng per spot. The mean value±SD of slope and intercept were 3.38 ± 1.47 and 986.9 ± 108.78 with respect to peak area. The method was validated for precision, recovery, and robustness. The limits of detection and quantification were 3.99 and 12.39 ng per spot, respectively. Cefpodoxime proxetil was subjected to acid and alkali hydrolysis, oxidation, and thermal degradation. The drug undergoes degradation under acidic and basic conditions, indicating that the drug is susceptible to both acid and base. The degraded product was well resolved from the pure drug, with significantly different R f value. Statistical analysis proves that the method is repeatable, selective, and accurate for the estimation of the investigated drug. Conclusion: The proposed HPTLC method can be applied for identification and quantitative determination of cefpodoxime proxetil in both bulk drug and pharmaceutical formulation.
  2,865 240 3
REVIEW ARTICLE
Embryonic stem cells: An alternative approach to developmental toxicity testing
S Tandon, S Jyoti
April-June 2012, 4(2):96-100
DOI:10.4103/0975-7406.94808  
Stem cells in the body have a unique ability to renew themselves and give rise to more specialized cell types having functional commitments. Under specified growth conditions, these cell types remain unspecialized but can be triggered to become specific cell type of the body such as heart, nerve, or skin cells. This ability of embryonic stem cells for directed differentiation makes it a prominent candidate as a screening tool in revealing safer and better drugs. In addition, genetic variations and birth defects caused by mutations and teratogens affecting early human development could also be studied on this basis. Moreover, replacement of animal testing is needed because it involves ethical, legal, and cost issues. Thus, there is a strong requirement for validated and reliable, if achievable, human stem cell-based developmental assays for pharmacological and toxicological screening.
  2,648 170 6
ORIGINAL ARTICLES
3D-QSAR studies on CCR2B receptor antagonists: Insight into the structural requirements of (R)-3-aminopyrrolidine series of molecules based on CoMFA/CoMSIA models
Swetha Gade, Shaik Mahmood
April-June 2012, 4(2):123-133
DOI:10.4103/0975-7406.94813  
Objective: Monocyte chemo attractant protein-1 (MCP-1) is a member of the CC-chemokine family and it selectively recruits leukocytes from the circulation to the site of inflammation through binding with the chemotactic cytokine receptor 2B (CCR2B). The recruitment and activation of selected populations of leukocytes is a key feature in a variety of inflammatory conditions. Thus MCP-1 receptor antagonist represents an attractive target for drug discovery. To understand the structural requirements that will lead to enhanced inhibitory potencies, we have carried out 3D-QSAR (quantitative structure-activity relationship) studies on (R)-3-aminopyrrolidine series of molecules as CCR2B receptor antagonists. Materials and Methods: Comparative molecular field analysis (CoMFA) and comparative molecular similarity indices analysis (CoMSIA) were performed on a series of (R)-3-aminopyrrolidine derivatives as antagonists of CCR2B receptor with Sybyl 6.7v. Results: We have derived statistically significant model from 37 molecules and validated it against an external test set of 13 compounds. The CoMFA model yielded a leave one out r 2 (r 2 loo ) of 0.847, non-cross-validated r 2 (r 2 ncv ) of 0.977, F value of 267.930, and bootstrapped r 2 (r 2 bs ) of 0.988. We have derived the standard error of prediction value of 0.367, standard error of estimate 0.141, and a reliable external predictivity, with a predictive r 2 (r 2 pred ) of 0.673. While the CoMSIA model yielded an r 2 loo of 0.719, r 2 ncv of 0.964,F value of 135.666, r 2 bs of 0.975, standard error of prediction of 0.512, standard error of estimate of 0.180, and an external predictivity with an r 2 pred of 0.611. These validation tests not only revealed the robustness of the models but also demonstrated that for our models r 2 pred, based on the mean activity of test set compounds can accurately estimate external predictivity. Conclusion: The QSAR model gave satisfactory statistical results in terms of q 2 and r 2 values. We analyzed the contour maps obtained, to study the activity trends of the molecules. We have tried to demonstrate structural features of compounds to account for the activity in terms of positively contributing physicochemical properties such as steric, electrostatic, hydrophobic, hydrogen bond donor, and acceptor fields. These contour plots identified several key features, which explain the wide range of activities. The results obtained from models offer important structural insight into designing novel CCR2B antagonists before their synthesis.
  2,715 82 -
EDITORIAL
Systemic delivery of biopharmaceuticals: Parenteral forever?
Ana Grenha
April-June 2012, 4(2):95-95
DOI:10.4103/0975-7406.94807  
  1,746 92 2
LETTERS
Periodontal therapy: A useful adjunct to improve glycemic control
Lata Goyal, ND Gupta, Afshan Bey
April-June 2012, 4(2):173-174
DOI:10.4103/0975-7406.94830  
  1,665 77 1
Essential medicines concept for quality assurance of health care facilities
Dixon Thomas, G Seetharam, Gerardo Alvarez-Uria
April-June 2012, 4(2):172-172
DOI:10.4103/0975-7406.94826  
  1,481 82 1
Rising incidence of high MIC for vancomycin among Staphylococcus aureus strains at a tertiary care hospital in South India
Banda Venkata Ramana, Abhijit Chaudhury
April-June 2012, 4(2):173-173
DOI:10.4103/0975-7406.94829  
  1,311 82 1
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