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Editor-in-Chief (Hon)

Dr Girish G

United Kingdom

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DrKLakshmi Narayana

India

Managing Editor

Prof Gopkumar P

India

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Dr Thawatchai PhaechamudThailand

Prof Jeanetta du PlessisSouth Africa

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DrAmrutha RadhakrishnanIndia

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India

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Dr Mahesh Kumar Gupta India

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Current Issue

RES J PHARM BIOL CHEM SCI

Volume 4 Issue 4 2013 (October - December)

1 Preliminary Physico-phytochemical Study and Pharmacognostical Standardization of Psidium guajava Leaves

Krishanu Samanta Romi Yadav Rishikant Tripathi Ashok Kumar and Emdad Hossain

2 Interaction Study of Curcumin with 1-Butanol Binary Mixture

Manjunath M S and J Sannappa

3 Formulation and Evaluation of Amlodipine Besylate Floating Tablets

Ramasubramaniyan P Palanichamy S Deepu VM and Rajesh M

4 Dried Blood Spot Sampling Analysis Recent Advanced and Applications

Jimi M Desai and Ravindra RP

5 A Report on Rubber Degrading Bacterial Sps from Vellore Soil Contaminated With Tyre Waste

KP Pramodh Kumar V Sai Shiva Shankar R Deepak Suneetha V and Bishwambhar Mishra

6 Synthesis and Characterization of Sebacoyl Bis-P-Bromophenyl Urea Polymer

Ghubde RS Bonde AD Gandhare NV and Juneja HD

7 Mercury Exposure Effects to Skin Tissue of Mus Muscullus at Fibroblasts Cell Proliferation and Collagen quantity

Titik Taufikurohmah Agoes Soegianto I Gusti Made Sanjaya Afaf Baktir and Achmad Syahrani

8 Study of the role of Copper Zinc and Magnesium in Diabetic Nephropathy

M Prasad Naidu Shiva Kumar S Mahaboob Vali Desai Madhav and G Subrahmanyam

9 Scarabiasis ndash A Rare Disease with a Rare Presentation

Rugmini Kamalammal and Soorya Rao R

10 Effect of Pioglitazone on Abdominal Fat Distribution in Type 2 Diabetic Patients

Gowri Thilagam T Tamilarasi SParameswari R Raadhika R and Mathivani M

11 Electrochemical behaviour of 2-hydroxy-5-methyl benzophenone benzoylhydrazone

D Manjula and P Venkata Ramana

12 Pattern of Use and Adverse Reactions to Antisnake Venom in Haemotoxic Snake Bite

Mathivani M Parameswari R Sarojini RGeetha K and Gowrithilagam T

13 Biosynthesis of Zirconium Oxide Nanoparticles using Marine Actinobacteria and its ApplicationsKV Bhaskara Rao Divyaa Srinivasan Shane Dan Christo Sruthi Laura George L Karthik and Gaurav Kumar

14 Preparation and Properties of Papain Immobilized onto Metal Ions Cross-linked Chitosan Beads

Sari Edi Cahyaningrum Narsito Sri Juari Santoso and Rudiana Agustini

15 Formulation Development and Evaluation of Orally Disintegrating Tablets of Losartan Potassium by Direct Compression Method

Jagadale Sachin K Patil Pradeep S and Navale Rajini




16 Evaluation of Memory Enhancement Activity and Shock Motivated Brightness Discrimination Response by using Y-Maze

D Eswar Tony T Vijaya D Sathish Kumar M Sathish Kumar A Narendra Babu and N Rama Rao

17 Prescribing Patterns of Antihypertensives in Pregnancy Induced Hypertension (PIH) in Relation with Food and DrugAdministration (FDA) Categories and Essential Drug List in Two Tertiary Care Hospitals Gulbarga

Tanuja V Hooli Sathisha Aithal and S Manjunath

18 Towards the Prediction of a Three-Dimensional Structure of Human GLP-1 Receptor

A Ranganadha Reddy Sreedhara R Voleti and Ch Lakshmi Padma

19 Effect of Substrate Temperature on Structural Optical and Surface Morphological Properties of Spray Deposited V2O5Thin Film

Mansur Bashaand L Akilasundari

20 Studies on Toxicity of Ag (I) on Plants and Microbes

Devlina Das Lakshmi V Nilanjana Das and Vimala R

21 Role Of Nano Encapsulation And Inclusion Complexation In Mouth Dissolving Strips Of Atorvastatin

M Abinaya D Ramya Devi and BN Vedha Hari

22 Risk Prediction of Morbidity and Mortality in Emergency Laparotomy by Possum Equation

Srinath S Naveen HSand Manjunath

23 DNA Binding and Antimicrobial Studies on Co (III) and Fe (II) Metal Complexes Containing Mixed LigandsSreekanth B Gopinath SM Veena Pillai V Ismail Shareef M Jagan Mohan Reddy VishnuvardhanTK Murali Krishna P and

Sridhara V

24 Treatment of Frozen Shoulder A Double Blind Study Ccomparing the Impact ofTriamcinolone Injection Alone or In Associationwith Joint Distention

Saeidian Seyed RezaForough Bijan Hemmati Ali Asghar and Tahmasebi Morteza

25 Development of New Analytical Method and Its Validation for the Determination of Loratadine in Bulk and MarketedFormulation

Megha J Solanki EVS Subrahmanyam and AR Shabaraya

26 Development of New Analytical Method and its Validation for the Determination of Metoclopramide Hydrochloride in Bulk andMarketed Formulation


27 Preliminary Phytochemical Investigation on A Few Cucurbitaceae Plants

Bhavani MB Leelavathi S and Ismail Shareef M

28 Superporous Hydrogel (SPH) An Innovative Approach of Gastro retention

Leena P Deore and Devidas G Bachhav

29 Clinical Study of Pulmonary Tuberculosis in Diabetes MellitusSharath Kumar D Shah Gireesh AS Mounika Kilariv Ghouse Pasha Anupam Biswas Neelesh Kumar S Shah Chandana SS and

Kanu Priya

30 Towards understanding Alzheimers Disease An Overview

Mayur Bagad Debajyoti Chowdhury and Zaved Ahmed Khan

31 Bacterial Profile and Antibiotic Susceptibility Pattern of Blood Culture Isolates from Pediatric Age Group Attending A TertiaryCare Centre

Bindu D Chitralekha S Menezes GA Illamani V

32 Thermo gravimetric Analysis of Copper (II) soaps Derived from Groundnut (Arachis hypogaea) and Sesame (Sesamumindicum) Edible Oils

Sharma R Heda LC Joram Aand Sharma

33 Investigation of In Vitro Anti-Inflammatory and In Vitro Anti-Oxidant Activity of Bark of Albizia procera Benth

Sangeetha M Chamundeeswari D Saravana Babu C Rose C and Gopal V

34 Acute Hemorrhagic Cystitis in Children A Case SeriesGopalakrishnan Manigandan Chandrasekaran Venkatesh Sadagopan Srinivasan Shyam Choudhury Puskar and Prasant

Nayak

35 Epigenetic Modulation Mechanisms in Psychiatric Disorders Gene and Trigger and Erase and Re-write Hypothesis

Akhil P Nair and Zaved Ahmed Khan

36 Antihyperlipidemic and Antiatherosclerotic Activity of Rimonabant in Wistar albino Rats

Ashish Mahajan Praful Patel and Patil SD

37 The Role of Serum Uric Acid in Acute Ischemic Stroke




Shashikala Lamani and HL Vishwanath

38 A Flexible Regulatory Approach for Different Raw Materials Suppliers Using QbD Principles

Buket Aksu Mehmet Aydogan Bayram Kanik and Ediz Aksoy

39 Prenatal Isotretinoin Exposure Reduces the Neuronal Population of Hippocampus in RatsJai Aditya Sampath Madhyastha Sudhanshu Sekhar Sahu Vasudha Saralaya Divya Premchandran Teresa Joy

40 Case Report A Rare Case of Primary Ovarian PregnancyHemant Deshpande Shilpa Chaudhari Chandrakantmadkar Charusheela Goreand Poorva Deshpande5

41 Coordination Possibility of Uracil and Applications of Some of Its Complexes A Review

Oladipo MAand Isola KT

42 Clinical Spectrum of Presentation in HIV Infected Children with Correlation to CD4 PercentagePrabhavathi R Basavaraj Veera Shankar M Taru Sharma Bhavya Vinodchandran Remya and Sreekantha

43 DNA Damage Assessment in Essential Hypertensive Patients Using the Single Cell Gel Electrophoresis (SCGE) Assay

Tarandeep Kaur and Gandhi G

44 The Effect of NF-Kappa B and Angiotensin II on the Proliferation in Human Breast Adenocancer Cell Line

Ozunal ZG Bayram R Yavuz MZ Ucbek A Aydogar A Uzun Ouml and Gepdiremen A

45 Free Radical Scavenging Capacity and Antioxidant Activity of an Ayurvedic Medicinal Plant Solanum rubrum Mill

Santhosh Kumar S Subramanian A SUJA SK Sudarshan M and Chakraborty A

46 Examination Screening and Statistical Analysis with respect to Prevalent Alzheimerrsquos Disease

Parul S Kamat and Suneetha Vuppu

47 Antibiotic Resistance in Food Poisoning Caused By Escherishia Coli O157H7 in Hospitalized Patients At 5 Years in Iran

Moghni M and Barati S

48 Development of New Analytical Method Validation for the Determination of Fluoxetine HCl in Bulk and Marketed Formulation byColorimetric Method

Bini Makadia EVS Subrahmanyam and Ramakrishna Shabaraya

49 Development of New Analytical Method Validation forthe Determination of Lamivudine in Bulk and marketed Formulation byColorimetric Method


50 Ameliorative Effect of Fish Oil on the Cisplatin Induced Hepatotoxicity and Nephrotoxicity in Rats

Mohamed SA El-Gerbed

51 Changing Spectrum of Antibiotic Sensitivity in Enteric Fever- A Six Year Retrospective Study in North India

Geetika Dheer Shaveta Kundra Atul Goel Tejinder Singh and Vandana Berry

52 Electrocautery versus Scalpel Incision in Inguinal Hernioplasty

Shekhar Upadhyay and Naval Bansal

53 Development of Validated Method for Determination of Residual Solvents in Guaifenesin and Imidazole Alcohol by GasChromatography (GCFID) with Head Space

Suresh Kumar Agrawal and Devendra Singh Rathore

54 Our Work on Capillary Zone Electrophoresis (CZE) and Micellar Electrokinetic Capillary Chromatography (MECC) under reviewSince 1989 and proposing a new mode of operation for anions experimentally called Reverse Direction Anion CapillaryElectrophoresis

KIUMARS GHOWSI and HOSEIN GHOWSI

55 RSM Mediated Optimization of Amylase Production from Marine Bacillus spVITRKHB

KV Bhaskara Rao H Bose K Richa K Singh L Karthik and G Kumar

56 Effect of Difference in the Dietary Galactose on the Rate of Onset and Progression of Cataract in RatsNurul Alimah Abdul Nasir Renu Agarwal Effat Omar Nor Salmah Bakar Sushil Vasudevan Renad Alyautdin and Nafeeza

Mohd Ismail

57 Formulation and Evaluation of Floating Drotaverine Hydrochloride Tablets Using Factorial Design

Om Prakash S Saraf M Rahman Neeraj Agnihotri and Vinay Pathak

58 Synthesis Spectroscopic and Thermal Studies of Newly Synthesized Transition Metal Coordination Polymers

SS Bhuyar HD Juneja and LJ Paliwal




59 Preparation Characterization of In-Situ Prepared SilverPVDF-PVP Nano-Composites

AM Abdelghany EM Abdelrazek AH Oraby and E Lasheen

60 An Epidemiologic Study of Gender Differences in Dental Space Anomalies

Mahesh Shivaji Taru and Kiran Plikundwar

61 Genotoxic Effect of Lead Acetate on Drosophila Melanogaster

MZ Fahira Reshman R Sumitha and V Deepa Parvathi

62 Structural and Electronic studies on Al Substituted SnO2 nano structures a DFT Study

Saravanakannan V and Radhakrishnan T

63 Evaluation of Elemental Profile of Tecomella undulata (Seem) An Endangered Medicinal Plant

Saraf Aparna and Sankhla Shweta

64 A Study on Similar Look like and Sound like Brand

Kiron SS PL Rajagopal Saritha M and Sreejith KR

65 OxidationndashReduction of methionine Sulfoxide Induced by an ArgonndashHydrogen Plasma Jet

Wada T Munegumi T and Harada K

66 Antibacterial Activity of Essential Oils of Chamaerops humilis (Arecaceae) on Some Pathogenic Bacteria

HASNAOUI Okkacha ADLI Djallal Edineand SENNOUR Raja

67 Synthesis and Characterization of Some Biologically Active Heterocycles Containing Nitrogen

Ratnesh Das and Gulzar Khan

68 Chemical Composition of the Essential Oils from Flowers Stems and Roots of Dorema ammoniacum DDon from Iran

Masoud Sadeghei Takallo Sami Sajjadifar and Maziar Mansouji Avval

69 Prevalence of Hypertension and Diabetes in Ischemic Stroke Patients in South Indian Population

Binu Mathew Abhilash Thomas and Suchandra Sen

70 Analysis of Stability of Granisetron Hydrochloride in Nasal Formulations by Stability-Indicating RP-HPLC Method

BhaleraoAV Shirolkar SV and Chitlange SS

71 Induced Biosurfactant Production and Degradation of Lindane by Soil Basidiomycetes Yeast Rhodotorula sp VITJzN03

Jaseetha Abdul Salam and Nilanjana Das

72 Chemical Weapons Lethal Weapons of Uncivilized World

Inbaraj SD and Menezes GA

73 Biopharmaceutical and Physicochemical Study of Substance and Suppositories with Tamsulosin Hydrochloride

Vita Gritsenko Olena Ruban Inna Kovalevskaya and Dennis Pulyaev

74 Visual Development and Visual Acuity Testing In Children

Kashinatha Shenoy M Gopalakrishna K and Preetha

75 A Review on Leucoderma

Sukhbir KaurHarminder Pal Singh Kahlon and Kushal Dhir

76 Ftir Analysis of Some Pills of Forensic Interest

Rajvinder Singh R Ajit Kumar and Ramanjeet Kaur

77 Impact of Noise Stress on the Immune Status of Albino Rats

Archana R

78 Synthesis Characterization Thermal Studies of Copper (II) Amino Acid Complexes

Shraddha Shukla Anupama Kashyap and Anil Kashyap

79 Diagnosis and Classification of Level of Kidney Function UsingAssociative Neural Network and Polynomial Neural Network

Rajalakshmi M Neelamegam P and Bharathi N

80 Impact of an Insecticide rsquoEncounterrsquo (Herbal Plant Extract) on Carbohydrate Content in the Freshwater Fish Labeo Rohita

Binukumari S and Vasanthi J

81 Antioxidant and Anti-inflammatory Activities of Flowers ofPlumeria rubra L f rubra and Plumeria rubra f lutea AComparative Study

Kalam Sirisha Yegnambatla Rajendra Periasamy GomathiKasarla Soujanya and N Yasmeen




82 Screening of Latex Producing Plants for Their Milk Clotting Activity

Anusha R Maheshwari Kumari Singh and Bindhu OS

83 Unascended Kidney A Case Report

Gireeshand Nagashree MV

84 Good Clinical Practice (GCP) and Its Importance

Anjula Sachan Pratap Shankar and Rakesh Kumar Dixit

85 Bacopa monniera Treatment Reverses Chronic Unpredictable Stress Induced Depressive like Behavior by IncreasingExpression of Neurotrophins in Rat Brain

Somoday Hazra Sourav KumarRitabrata Banerjee Rudra Prasanna Banerjee and Amal Ch Mondal

86 Diversity and Distribution of Pteridophytes in the Flora of Ethiopia and Eretria

Fikre Dessalegn

87 Amelioration of Histopathological Effect of Lead Induced Testicular Damage by Vernonia Amygdalina Leaves

Ligha AEKola-Ojo OB and Jegede AE

88 Study of Probiotic and Antioxidant activity of Lactobacillus sp

Pradeep Kumar Sharma Rajdeep Roy Mythili Sathiavelu and Sathiavelu Arunachalam

89 Influence of Formulation and Process Variables on the Formation of Rifampicin Nanoparticles by Ionic Gelation Technique

Subashini Rajaram and Rajendran Natham

90 A Review Neurocysticercosis Diagnosis and Treatment Issue

Ayush Dubey Somnath Singh Raghuvanshi TN Dubey

91 Bran Bread Chemical Composition Fungal Load Biological Impacts and Intervention

Sherif EA Badr Hanan MA El Ghandour Eman M Ragheb and AS Abdel Rahman

92 Knowledge on Human Papilloma Virus Vaccination

Vineeta Jose Malathi G Nayak and Preethy Jawahar

93 Antagonistic Effect of Benzalkonium Chloride on Nicotine Induced ContractionsBhaskara Reddy Nallamilli Sucharita P Lakshmi Prasanna M Maruthi V Ramakrishna K Ravi Kishore B Satyanarayana S

94 Synthesis Characterisation and Screening of Anthelmintic Activity of Some Novel Schiff Bases

Sharmila Sutradhar Amtul Muneem Maliha and Ayeesha Humera

95 Antioxidant and Anti-Inflammatory Properties of Autolysed Extract of the Indian Earthworm Pheretima posthuma afterPreliminary Purification - An In Vitro Study

Mahendra Kumar Verma and Kota Sobha

96 Pharmacognostical Phytochemical and Antibacterial Evaluation of Berberis Tinctoria Lesch (Stem Wood and Stem Bark)

Saha Pradeep and Patel Kanu Bhai Ramesh Bhai

97 A Facile and Efficient Method for the Synthesis of Quinoxaline Derivatives Using [2-(Sulfooxy)Ethyl]Sulfamic Acid as a Novel

Difunctional Bronsted Acid Recyclable and Organocatalyst

Sami Sajjadifar Hadi Noorizadeh Hojat Veisi Omid Louie Maziar Mansouji Avval Sobhan Rezayati

98 RecentPharmacologicalReviewon Cinnamomum tamala

Borhade Pravin Lone Krishnkant Joshi Shreyas Kadam Ajay and Gaikwad Priyanka

99 Study of Utilization of Antenatal Care Services in Slum and Non- Slum Areas of Vijayawada City

Swetha R J Ravikumar and R Nageswara Rao

100 Vinca roseaNormalizes Oxidative Stress and Inhibits Hyperglycemia Induced Increase in VEGF in Zebrafish RetinaJayshree Nellore Cynthia Pauline P Surya Prabha MohananRamya Ravikumar and Ragavi Chidambarathanu Pillai

101 Recent Progress in Fiber Optic Biosensors Applications

C Bosch Ojeda and F Saacutenchez Rojas

102 Nasal Drug Delivery System as a Potential for Nasal Solution of Metoclopramide ydrochloride ndash In Vitro and In Vivo Properties

Menaka M and Pandey VP

103 Biology of Skin Aging A Review

L Chandersekar



L Chandersekar

104 Spectroscopic Investigation of Optical Parameters of Pure PMMA and Azo - dye Doped PMMA films

HM Zidan NA El-Ghamaz AM Abdelghany and A Lotfy

105 Validated Spectrophotometric Methods for the Determination of Nabumetone in Tablets Dosage Form Using ThreeDinitrobenzene Reagents

Mona M Bedair Azza A Gazy Tarek S Belal Karin M Guirguis

106 Nutrient Recovery from Sewage Wastewater and Bittern as Precipitated Struvite Using Zeolite and Activated Carbon asAdsorbent

Sh El Rafie Randa Othman Marwa M Shalaby and S Hawash

107 Auditing Waste Management Practices in an Indian City

Ashutosh Das R Vasanthi Mukesh Goel and K Ravi Kumar

108 Modeling of Freeboard Fluidized Bed Gasifier Integrated to Bubbling Dense Bed

A Venkata Narayana and M Vijaya Leela

109 Protein Modification during Germination of Sorghum Bicolor

Malomo Olu and Alamu E A

110 Salivarius K12 as A Probable Probiotic

Sheen Ann John B M Shantala and V Narashima Rao

111 Synthesis Spectroscopic and Thermal Investigation of New Nickel (II) Amino Acid Complexes


112 Performance of an Anaerobic Stage Reactor (ASR) Treating Synthetic Wastewater during Start-Up Phase Using Palm Oil MillEffluent (POME) Sludge

Mahat SB Chelliapan S Yuzir A Md Din MF Anwar AN Othman N and Shamsuddin S

113 Relation between Waist-Hip Ratio and Lipid Profile in Female Type 2 Diabetes Mellitus atients

Narasimha Rai K and Jeganthan PS

114 Spectrophotometric Determination of Drugs Using 23-Dichloro 56-dicyano p- benzoquinone as Analytical Reagent

Sayaji Rao

115 The Optimization ofSodiumCarboxymethylCellulose(NA-CMC) Synthesizedfrom Water Hyacinth(Eichhornia crassipes (Mart)Solm) Cellulose

Ida Musfiroh and Aliya Nur Hasanah Iman Budiman

116 PREs-Clustered motifs in Drosophila melanogaster

Sabahuddin Ahmad Abuzar Hamza and Khalid Raza

117 The Effect of Subculture on the Bacoside A Content in Adventitious Shoot Cultures of Bacopa monnieri (L)

PM Naik BR Patil LG Jaggal and VK Jangid

118 Synthesis and oxidative dehydrogenation of 3 4-dihydropyrimidin-2-(1H)-one by Ce (SO4)24H2O

SK Pardeshi D D Kumbhar B Y Waghamare and PD Lokhande

119 Nootropic Activity of Zingiber Officinale in Albino Mice A Behavioral and Neurochemical Approach

Abhisek Pal Monalisa Jena Swati Mishra

120 Studies on Comparative Larvicidal Efficacy of Methanol Extracted Latex of Calotropis Procera and Temephos against AedesAegypti in Arid Parts of Rajasthan

Manju Singhi And Anil Purohit

121 Phosphate Adsorption on Sediments from Rhumel and Boumerzoug Rivers in the Northeast of Algeria

Sarah Azzouz and Chahrazed Boukhalfa

122 Testicular Effects of Artemisinins Are They Reversible

Jonah Sydney Aprioku

123 Comparative Smooth Muscle Relaxant Activity Of Dihydropyrimidine Derivatives 5-Acyl-6-Methyl-4-Phenyl-2-S-Ethyl-14-Dihydropyrimidine (BK VI) 5-Acyl-6-Methyl-4(23 Methylenedioxy) Phenyl 2-S-Benzyl-14-Dihydropyrimidine (BK VII) andNifedipine on Isolated Rat Uterus

Shalini Salwan Poonam Salwan Walia R Bajaj V K And Kaur B

124 Practical Method for Isolation of Residuals Determined from Polynomial Fitting to Gravity Data (Case Study Bandar CharakHormuzgan Iran)

Reza Toushmalani and Amir Esmaeili

125 Confirmation Of Quantity Of Inactive Gradients Added In The Pharma Drugs By Determining Mass Attenuation Coefficient

Manjunath Aand Kerur BR




126 Phytochemical and Anti-inflammatory Activities of the Methanol extract and Fractions of Dalbergiella welwitschii Baker (Bakerf) leaves

Fred-Jaiyesimi Adediwura and Akinla Oluwakemi

127 Nutrient Intakes and Digestibility in Dairy Calves Fed Congo-Signal (Brachiaria Ruziziensis) Based Complete Feed Block inMizoram

R Buragohain P Saikia and H Bayan

128 Cumulative Effects of Septic System Disposal and Evolution of Nitrate Contamination Impact on Coastal Groundwater inTuticorin South Tamilnadu India

S Selvam G Manimaran and P Sivasubramanian

129 Clostridium Difficile Infection An Overview of the Disease and Its Pathogenesis Diagnosis Treatment Prevention AndManagement

Samir Issa Bloukh

130 Dental Caries Experience among Haemophilic Children

Mythri H Jagdish G R and Chandu G N

131 Estimation of Serum Nicotine Levels among Tobacco Users

Sujatha S Reddy and Radha Prashanth

132 Process Development for Lipase Extraction and the Effect of Extracted Lipase on Triglyceride Base System

Sasikan Kupongsak and Pattama Lucharit

133 Anaerobic Biotechnology for Pharmaceutical Wastewater Treatment

Chelliapan S and Sallis PJ

134 Prevalence and Antibiotic Susceptibility Pattern of Methicillin Resistant Staphylococcus aureus (MRSA) in Pus Samples in aTertiary Care Hospital

SandhiyaR Lakshmipriya R and Esthermary

135 Schizophrenia Interaction between Dopamine Serotonin Glutamate GABA and Norepinephrine

Senthilkumaran Jagadeesh J and Shalini Natarajan

136 Seroprevalenceof HBs Ag and HCV in Healthy Blood Donors at a Tertiary Care Hospital in India

Kanchan Bhardwaj Prasad KM Ravisha Bhardwaj and BL Bhardwaj

137 Purification of Beta Asarone from Acorus Calamus L

S Asha Devi Subhasini and S Babu

138 Synthesis Characterization Antimicrobial Investigations of Copper (II) Complexes with Some Benzylbenzimidazole Derivatives

Sunita B Garud and L P Shinde

139 Role of Inflammation in Growth Invasion and Metastasis of Oral Squamous Cell Carcinoma- A Molecular Insight

Shyamala K Sanjay Murgod and Girish HC

140 A comparative study of solubility enhancement of enalapril using formulation of solid dispersion and using hydrotropicsolubilization technique

Dahima R and Gangwal S

141 Contributions to the environmentof priority Polynuclear Aromatic Hydrocarbonsfrom the Coal Camp Mechanic Village (CCMV) inEnugu Nigeria

Dike Henry OGBUAGU and Kingsley Nnaemeka OGBONNA

142 Climacteric Symptoms and Health Status of Menopausal Women - North Karnataka

Chandramati J Rokhade Dhiraj J Trivedi and Khyrunnisa Begum

143 Antioxidant and Anti-inflammatory Activities of Agricultural By-products and Medicinal Herbs As Potential Functional AnimalProducts

Sunhee Cheong Byongtae Jeon Eunkyoung Kim PyojamPark Mirae Oh and and Sangho Moon

144 An Exciting Style of Teaching to Increase the Exam Score of Students

S Roopa Bagavad Geetha and Anitha Rani

145 16S rDNA Based Identification of Alkaline Protease Producing Alkaliphilic Bacillus Sp Isolated From Dairy Industry Soil AndEvaluation of the Enzyme Potential In Detergent Formulation

Jyoti Verma Sangeeta Saxena and Shikha

146 Novel Synthesis and Anti-tumour Activity of 2-Hydrazino-1H-benzimidazoles

Sh M Abu-Bakr Kh M Abu-ZiedMahmoud Youns Amel Hashim and Hoda I El-Diwan



147 Phylogenetic Relationships of Selected Kaempferia Plants in Thailand Based on RAPD MarkerOrawan Theanphong Witchuda Thanakijcharoenpath Nijsiri Ruangrungsi Chanida Palanuvej and Kanchana Rungsihirunrat

148 Synthesis and Evaluation of New Novel Heterocycles Containing Benzothiazoles (1 3) for Pharmacological Screening

Ashok Kumar KV B Gopalakrishna E Jayachandran Ramkrushna and Babarao Sawandkar

149 Development and Validation of an RP-HPLC method for the estimation of Tolterodine in Raw materials and Tablet dosage forms

Vijaya Bhaskar S Seshagiri Rao JVLN and Vijaya Sree

150 Development and Validation of an RP-HPLC Method for the Estimation of Trapidil in Raw Materials and Tablet Dosage Forms

Vijaya Bhaskar S Seshagiri Rao JVLN and Seetha Ramaiah P

151 Screening Identification and Fatty Acid Composition Analysis of Mercury Resistance Microalgae from West SumateraIndonesia

RL Amza K Oh-Hashi Y Yamaguchi K Tanaka S Yoshida A Dharma E Munaf and M Koketsu

152 Investigation of Massage and Moxibustion Treatment Efficacy for 270 Cases of Waist and Haunch Pain

Li Cheng and Zhao Chang-Li

153 Microbiological Characteristics of Sour-Milk Feed Supplements and their Influence on Intestinal Micro-Biocenosis of Piglets

O Vichko V Chervetsova and V Novikov

154 In Vitro Anti Oxidant Activity of Extracts and Stigmasterol from Leaves of Clerodendrum inerme Linn

B Shanthakumar M Sathish and A Jerad Suresh

155 In Vitro Antioxidant Activity and Phenolic Contents of the Leaves of Olax subscorpioidea and Distemonanthus benthamianusKouassi Konan Nrsquoguessan Jean David Meacuteiteacute Souleymane Yapi Ahoua Yapi Houphoueumlt Feacutelix Djaman Allico Joseph

156 The Effect of Sintering Process on the Characteristics of Hydroxyapatite from Cuttlefish Bone (Sepia Sp)

Aminatun Siswanto YM Penga Istifarah and R Apsari

157 Prediction of Pb (II) and Cu (II) Ions Biosorption by Annona muricata L Seeds Using Artificial Neural Network (ANN) Approach

Zulfi Abdullah Melia Innes Kurniawan Rahmiana Zein Hermansyah Aziz and Edison Munaf

158 A Prevalence Study of Visual Impairment and Blindness among Elderly in an Urban Slum Community of Pune CantonmentIndia

R Harnagle and PS Chawla

159 Isolation Oil Producing Micro algae chlamydomonas snowii from Tropical Fresh Water Indonesia

Afny Varitha Abdi Dharma Edison Munaf Nasril Nasir and Afrizal

160 Synthesis of New Fused Tricyclic Quinoid Systems and Studying of Their Biological Activity In-SilicoYuliia Dumanska Yurii Shakh Anastasiia Kudrinetska Khrystyna Bolibrukh Andrii Karkhut Bohdan Lytvyn Oleksii Kovalchuk

Olha Marshalok Mykola Platonov Sviatoslav Polovkovych and Volodymyr Novikov

161 Potential Products of Coconut Shell Wood Vinegar

Ervina Mela Yandra Arkeman Erliza Noor and Noerazam Achsani

162 Influence of Cement Dust on Ceramic Properties of Basalt Bricks

Randa M Osman H Abu-El-Naga EA El-Alfi

163 RESIN BASED ROOT CANAL SEALERS AN OVERVIEW

Sandeep Thakur Mithra N Hegde and Darshana Devadiga

164 Potential Drug Candidates for Fast Dissolving Drug Delivery - A Review

K Kavitha Kumutha Subramaniam Boey Jia Hui K Santhi SA Dhanaraj and M Rupesh Kumar

165 Lasers in Restorative Dentistry An Overview

Suvidh Virmani Mithra N Hegde and Chitaranjan Shetty

166 Preparation Characterization 1H 13C NMR Study and Antibacterial Studies of Schiff Bases and Their Zn (II) Chelates

V Prakash and MS Suresh

167 A Study on the Pattern of Self-medication for Acne Vulgaris in MedicalParamedical Students

Jyothi R Deepa R Pundarikaksha HP and Girish K

168 In Vitro Antitumor Activity of Alcoholic Extract ofPiper Betel Leaf

VA Kangralkar and AR Kulkarni



169 A Prevalence Study of Dental Caries in Different Socio- Economic Status Among Clientele of A Tertiary Dental Centre atJabalpur India

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170 Ecotoxicological Studies on Heavy Metal Tolerant Microbes Isolated From Marine Ecosystem

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171 Synthesis and Biological Evaluation of Substituted Imidazo [21-b]-134-Thiadiazole Derivatives as Anti-Inflammatory Agents

Ramjith US Karki S S and Cyril Mathews Jacob

172 Preparation and Characterization of Sustained Release Tablet Containing Solid Dispersion Granules of an Anti-HypertensiveDrug

G Nisha Shetty and T Mukesh

173 Microalgae Application for Treatment of Textile Effluents

M Jaya Chitra Ashutosh Das Mukesh Goel and K Ravi Kumar

174 Primary MALT Lymphoma of Appendix A Rare Case Report

Sharique Ahmad Nishi Tandon Ruchi Dinkar Sonal Dhingra and Nirupama Lal

175 Molecular Line Probe Assay Genotype MTBDR plus for Rapid Detection of Primary Drug Resistance in Mycobacteriumtuberculosis Isolates From A Tertiary Care Hospital in Western India

Patil Seema Angadi Kalpana Modak Meera and Bodhankar MG

176 Studies on Novel Azetidinone and Their Biological Studies

Jigar D Bhatt Kiran S Nimavat and Kartik B Vyas

177 Hydrothermal Synthesis of MnO2 Nanoparticles using Teflon Lined Autoclave

M Senthilkumar Balamurugan and BG Jeyaprakash

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179 Role of Thymidine Kinase Gene in White Spot Syndrome Virus

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180 Amperometric Trace Determination of Se (IV) and Se (VI) using Thioglycolic Acid

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181 Anthelmintic Activity of Alcoholic and Aqueous Extract of Anisochiluscarnosus (Wall)

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ISSN 0975-8585

October-December 2013 RJPBCS Volume 4 Issue 4 Page No 120

Research Journal of Pharmaceutical Biological and Chemical

Sciences

Preparation and Properties of Papain Immobilized onto Metal Ions Cross-linked Chitosan Beads

Sari Edi Cahyaningrum1 Narsito2 Sri Juari Santoso2 and Rudiana Agustini1

1

Department of Chemistry Surabaya State University Indonesia 2 Department of Chemistry Gadjah Mada University Indonesia

ABSTRACT

Chitosan beads were prepared by using a cross-linking agent Cu(II) and Zn(II) metal ions and the metal ion- chitosan beads were employed in papain immobilization processes Studies on free and immobilized papain systems for determination of optimum pH optimum temperature thermal stability and reusability were carried out The results showed that free papain have been optimum pH 65 and optimum temperature 55

oC while the

immobile papain had optimum pH 8 and optimum temperature 85 oC The thermal stability of the immobilized

papain relative to that of the free papain was markedly increased The residual activity of papain immobilized on chitosan bead- metal ion was about 25 after 12 cycles of batch operation Keywords papain immobilization chitosan beads metal ions Corresponding author

ISSN 0975-8585


INTRODUCTION

Enzymes have a wide variety of biotechnological biomedical and pharmaceutical applications They are used as biosensors in bioengineering clinically as therapeutic agents in modern diagnostic tool and as catalyst for chemical and biochemical reactions A large research of work has been devoted to the polymeric carriers especially to immobilization of the proteins onto carriers [1-4] Since the recovery yield and reusability of free enzymes as industrial catalysts are quite limited attention has been paid to enzyme immobilization which may offer advantages over free enzymes for example possibility of continuous process controlled product formation ease of enzyme removal from the reaction mixture and adaptability to various engineering designs

Chitosan a poly-N-acetyglucosamine is a transformed oligosaccharide obtained by deacetylation of chitin and it is the second most abundant biopolymer after cellulose [24] Chitosan exhibits a unique set of characteristics such as biocompatibility biodegradability nontoxicity hydrophilicity remarkable affinity to proteins and high mechanical strength These characteristics make chitosan as desirable biomaterial for enzyme immobilization [24510] It can provide an optimal microenvironment for the immobilized to maintain relatively high biological activity and stability Increasingly over the last decade chitosanndashbased biomaterials were employed as enzyme immobilization in the form beads and membranes [67]

In this study chitosan beads were crosslinked with Zn(II) and Cu(II) for matrix papain

immobilization Various attempts have been made to stabilize papain for a more efficient use Papain and other proteolytic enzymes have been immobilized by radiation polymerization of various monomers [816] Covalent coupling of papain has also been shown in different studies performed by several workers [91112] However the biomatrices with entrapped enzymes tend to leak proteins with time This resulted in the activity losses as well as contamination of the product with the enzymes which is not acceptable for pharmaceutical applications The covalent coupling of enzyme can produce a considerable loss of activity due to the influence of the coupling conditions and to conformational changes in enzyme structure However irreversible binding of enzyme to the carrier during covalent coupling does not allow the recovery of the carrier from the carrier-enzyme complex [111314] A method is therefore needed in which the carrier should be easily regenerated and reused without reducing the immobilization yield Attempts have been made in this direction and a metal chelate regenerable carrier has been used to immobilize the papain This immobilization is based on the ability of protein side chains of cysteine histidine and tryptophan to substitute weakly bonded ligands in the metal complexes This method has a big potential and may be more versatile since it allows a selection among many chelating metal ions

MATERIALS AND METHODS

Papain (EC 34222) and Casein were obtain from Sigma Chem Co (st Louis USA) Chitosan was obtained from shell of shrimp with Meyer Methods (1989) All other chemicals were of analytical grade

ISSN 0975-8585


Preparation of Swollen Chitosan Beads

To prepare highly swollen beads an amount of chitosan flakes (1g) was completely dissolved in 01l of 1-moll acetic acid The resulting solution was sprayed into 125 of ml deionized water containing 15 g NaOH and 25 ml of 95 ethanol through a nozzle (12 mm diameter) The chitosan beads were swelled and washed with deionized water until the solution became neutral The diameter of wet beads approximately 23 mm The BET surface area of swollen beads was not measured because the drying was difficult The morphology of chitosan beads was analysis with SEM Determination of Immobilization Papain

The protein content of the chitosan-papain conjugate was calculated by subtracting the amount of protein determined in the centrifuged and washings following immobilization from the amount of papain used for immobilization The papain in the solutions was determined by the Bradford method [13] Immobilization Papain in Different pH

Papain was dissolved in 15 mM PBS with pH ranging from 40 to 80 respectively Each kind of chitosan beads-Zn(II) was incubated individually with above papain solution and shaken in a vibration for 12 h After equilibration the pH of each solution was detected with a pH meter and adjusted to certain value with PBS The adsorption capacity of papain in different condition was calculated by following equation Activity Assay of Free and Immobilized Papain

The activity of soluble papain was determined by the method of Kunitz as described by others using casein as substrate at 37ordmC and pH 82 The enzyme activity of immobilized papain was determined in a similar manner except that the reaction mixture was continuously stirred during the reaction One unit of enzyme activity is the amount of enzyme which produces TCA soluble peptides or amino acids giving a blue color equivalent to that of 05 mg tyrosine per minute at 37ordmC

Effect of pH and Temperature on the Papain Activity

The pH stabilities of the free and immobilized papain were by immersing the sample in PBS 15 mM in the pH range 4-10 Their thermal stabilities were assay by a standard activity assay in the themperature 40 to 90 oC

ISSN 0975-8585


Reusability of Immobilized Papain

For the reusability after each reaction run the immobilized papain was removed and wash with water to remove any residual substrate on the matrix It was then reintroduced into fresh reaction medium and enzyme activity was assayed at optimum condition

RESULTS AND DISCUSSION

Immobilization Efficiency in Different pH

The effect of pH on the adsorption of papain onto chitosan beads-Zn and chitosan beads-Cu chelated could be found that the maximal immobilized capacity onto matrix in pH 80 The decrease in the papain adsorption capacity in more acidic and more alkaline regions could be attributed to electrostatic repulsion effects between the opposite changed groups Proteins have no net charge at their isoelectric points and therefore the maximum adsorption from aqueous solutions is usually observed at their isoelectric points The isoelectric pH papain is 875 Papain immobilized on chitosan flake found that maximal immobilized capacity onto chitosan flake in pH 75 [13]

Picture 1 Effect of pH value on papain immobilization

However in the present study the maximum immobilization was not at this pH but had

slighty shifted toward more neutral pH values This could be due to preferential interactions between molecules and metal ion incorporated in polymeric matrix at neutral pH So in the following experiment the adsorption of papain on matrix was conducted in pH 80 Chitosan do not carry a charge at neutral pH If the pH value decrease in solution chitosan beads can be charged positively at lower pH because hydrogen ions can bind to free amino groups However limited number of available amino groups on cross-linked chitosan beads reduces number of bound hydrogen ions

ISSN 0975-8585



The pH dependence of the immobilized papain activity was compared with that of the free enzyme for casein in the pH range of 40-90 at 65 oC It can be seen from Fig 1 that optimum pH for the immobilized papain shifted slighty from 65 to 80 when compared with free one The relative activity of the papain immobilized was improved in a broad pH range compared with the free one The immobilization of enzymes to charged supports often leads to displacements in the immobilized enzyme and the bulk phase due the electrostatic interactions with the matrix

Picture 2 Effect of pH on papain activity

The temperature dependence of the activities of the free and immobilized papain was

studied in 15 mM PBS at temperature range 40-100 oC and temperature profiles of free and immobilized papain shown in Fig2 The optimum temperature range for free and immobilized papain was found to be about 50-60 and 75-85 oC respectively The conformational flexibility of the papain was affected by immobilization The immobilization of papain on chitosan beads with Zn(II) bifunctional agent caused an increase in papain rigidity which is commonly reflected by increase in stability towards denaturation by raising the temperature[813]

Picture 3 Effect of temperature on papain activity

ISSN 0975-8585


The thermal stability of immobilized papain was markedly increased relative to that of the native enzyme The thermal stability pf chitosan beads-Zn(II) papain at 80oC was improved dramatically

Picture 4 Thermal stability of papain


To investigate the reusability the enzyme-immobilized chitosan beads-Zn(II) was

washed with deionized water after one catalysis run and reintrodused into a casein solution for another hydrolysis Fig 4 shows the effect of repeated use on the activity of the immobilized papain It can be seen that the activity of the immobilized papain decay with recycled The residual activity of papain immobilized on chitosan beads-Zn(II) was about 25 after 12 cycles of bath operation The activity loss could be related to the inactivation of the enzyme caused by the denaturation of the protein and the leakage of protein and metal ions from the supportrsquos surface

CONCLUSIONS

The main advantage of immobilization papain on chitosan beads-Zn and chitosan beads-Cu consists its simplicity university stability and cheapness In this study a novel metal immobilized adsorbent was prepared Papain could be directly immobilized on the prepared chitosan beads-Zn and chitosan beads-Cu Papain immobilized has high stability and activity retaining

REFERENCES

[1] Betigeri SS Neau SH Biomater 2002 233627-36 [2] Chandy T amp Scharma CP Biomater 1990 18 1ndash24 [3] Chiou SH amp Wu WT Biomater 2004 25 197ndash204 [4] Dutta PK Dutta J Chattopadhyaya MC Tripathi VS J Polym Mater 2004 21321ndash333 [5] Huckel M Wirth HJ and Hearn MT J Biochem Biophy Meth 1996 31165-179

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[6] Jianmin Wu Mingming Luan Jiayin Zhao Int J Biol Macromol 2006 39185ndash191 [7] Kennedy JF amp Melo EHM Chem Eng Prog 1990 81-89 [8] Kiling Ali Onal Secil and Telepong Azmi Turk J Chem 2002 26 311-316 [9] Kise H and Hayakawa A Enz Microbial Technol 1991 13 584-588 [10] Krajewska B Enz Microbial Technol 2004 35 126ndash39 [11] Kumar M React Funct Polym 2000 46 1ndash27 [12] Kumakura M and Kaetsu I Int J Biochem 1994 161159-1161 [13] Liang Ding Zihua Yao Tong Li Qiang Yue Jia Chai Turk J Chem 2003 27 627- 637 [14] Li J Du YM Sun LP Liang HB Feng T Wei YN J Appl Polym Sci 2006 1013743ndash3750 [15] Pereira EB De Castro HF De Moraes FF Zanin GM K Appl Biochem Biotech 2001 91

739-52 [16] Sarah Afaq Electronic J Biotechology Catolica de Velparaaiso Chile 2001 6 234-237

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