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Volume 11 (2020) - Issue 5, May
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INTERNATIONAL JOURNAL OF PHARMACEUTICAL SCIENCES AND RESEARCHA Web of Science - ESCI Indexed Journal
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Dr. Shashi AlokAssistant Professor, Institute ofPharmacy, Bundelkhand UniversityJhansi (U.P.), India
Mrs. Monika SabharwalManaging Editor, International Journal ofPharmaceutical Sciences and Research,Panchkula (HR), India
Dr. Raymond. C. JagessarDepartment of Chemistry, University ofGuyana Faculty of Natural Sciences,South America
Prof. (Dr) G. K. DashFaculty of Pharmacy & Health Science,University Kuala Lumpur, Perak ,Malaysia
Prof. (Dr) J.O.C. Silva JuniorSchool of Pharmacy, Federal Universityof Para Belem, Para, Brazil
Prof. (Dr) Mohamed Abdel HamidProfessor of Pharmaceutical Chemistry &Former Dean of Faculty of Pharmacy, AinShams University, Cairo, Egypt
Dr. Narazah Mohd YusoffAdvanced Medical and Dental Institute,University Sains Malaysia KepalaBatas, Pulau Pinang, Malaysia
Dr. Sandeep ChaudharyInstitute of Microbial Chemistry MicrobialChemistry Research Foundation,Kamiosaki, Shinagawa-Ku, Tokyo, Japan
Dr. Mayank ThakurCharite University of Medicine, BenjaminFranklin Campus, Hindenburgdamm,Berlin, Germany
Dr. Satya Dev SharmaCancer Research Scientist,R.P.C.I.,Buffalo, New York, USA
Dr. S.H. YulianiFaculty of Pharmacy, Gadjah MadaUniversity, Sekip Utara Yogyakarta,Indonesia
Dr. Havagiray R. ChitmeProfessor, Oman Medical CollegeMuscat, Sultanate of Oman
Dr. Mohd. Sohail AkhtarDepartment of Pharmacognosy, Schoolof Pharmacy, University of Nizwa,Sultanate of Oman
Dr. S. PalaniSchool of pharmacy, 7th Octoberuniversity, Misruta, Libya
Dr. Nisha JoshephSchool of Pharmacy, College of HealthSciences, Mekelle University, Mekelle,Ethiopia
Dr. Roman PaduchMaria Curie- Sklodowska University,Lublin, Poland
Md. Hemayet HossainSenior Scientific Officer ChemicalResearch Division , (BCSIR), Dhaka,Bangladesh
Editorial Board
EDITOR-IN-CHIEF
MANAGING EDITOR
INTERNATIONAL ADVISORY BOARD MEMBERS
5/5/2020 Editorial Board | INTERNATIONAL JOURNAL OF PHARMACEUTICAL SCIENCES AND RESEARCH
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Prof (Dr.) Lawrence H. BlockDivision of Pharmaceutical Sciences 437Mellon Hall, Duquesne UniversityPittsburgh, PA
Dr. Rakesh TekadeUniversity of Central Lancashire Preston,England, UK
Gaurav SharmaDepartment of Pharmaceutical Sciences,Idaho State University Pocatello, USA
Dr. M.A.M OshiFaculty of Pharmacy, Omdurman IslamicUniversity, Omdurman, Sudan
Dr. Md. A. A. El Aziz Aly El DegwyFormulation section head, MepacoPharmaceutical Company, Maadi, Cairo,Egypt
Dr. Shankar JothiFacuty of Science, Technology & Engg.La Trobe University, Australia
Dr. Siddharthya MujumdarBoehringer Ingelheim PharmaceuticalsInc, Ridgebury Rd, Ridgefield, CT, USA
Dr. Rajesh G KatareDepartment of Physiology Otago Schoolof Medical Sciences University of Otago,New Zealand
Mr. Nipun MahajanSchool of Chemistry National Universityof Ireland Galway, Ireland
Dr. Prabhakar Reddy PolepallyInstitute of Pharmaceutical SciencesUniversity of Mississippi, MS, USA
Dr. Shruti RawalPostdoctoral Fellow, NYU LangoneMedical Center, New York, USA
Dr. Prasoon GuptaScientist, Florida Atlantic University BocaRaton, FL ,USA
Dr. Shahenda M. El-MesseryDepartment of Pharmaceutical organicChemistry, Mansoura University,Mansoura, Egypt
Dr. Shivanand PuthliPrincipal Scientist, TrisPharma Inc. , NewJersey, USA.
Dr. Ravi S ShuklaScientist III, Amneal PharmaceuticalsLLC, Brookhaven, New York, USA
Dr. Prity TomarLondon, United Kingdom
Dr. Shivkanya FuloriaFaculty of Pharmcy, AIMST University.Semeling Campus, Bedong. Kedah DarulAman, Malyasia
Dr. Mohammad Reza AbediDepartment of Applied Chemistry IslamicAzad University, Quchan Branch (IAUQ),Quchan, Khorasan razavi, Iran.
Dr. Srinivasan ShanmugamProject Leader, Formulation R&D, HanmiPharm. Co. Ltd., Hwasung-Si, Gyeonggi-Do, South Korea
Dr. Sunita SubramanianRollins Research Centre, Department ofBiochemistry, Emory University School ofMedicine, Atlanta, GA, USA
Dr. Bhagwat PrasadDepartment of Pharmaceutics, Universityof Washington Seattle, Washington, USA
Dr. Surendra Kumar JainNational Center for Natural ProductsResearch School of Pharmacy,University of Mississippi University, USA.
Dr. Pushkar M. KulkarniDepartment of Pharmaceutical Sciences,Bouve College of Health Sciences,Northeastern University, Boston, MA,USA
Dr. Monika SharmaPostdoctoral Fellow, NYU LangoneMedical Center, New York, USA
Sameer SachdevaResearch Scientist II, AmnealPharmaceuticals, NJ, USA
Dr. Ahmed S. ZidanCenter of drug evaluation and research,Food and Drug Administration, USA
Dr. Anekant JainUnited States of America
Dr. Animikh RaySchool of Pharmacy, University ofMissouri Kansas City (UMKC), USA
Dr. Javad Sharifi RadDepartment of Pharmacognosy Facultyof Pharmacy Zabol University of MedicalSciences Zabol, Iran
Dr. Chellappan Dinesh KumarDepartment of Life Sciences, School ofPharmacy, International MedicalUniversity, Kuala Lumpur, Malaysia
Dr. Ajaykumar N. SharmaThe University of Texas Health ScienceCenter, Houston, Texas, USA
Dr. Chin Jin HanDepartment of PharmaceuticalChemistry, Faculty of PharmaceuticalSciences, UCSI University, Cheras,Kuala Lumpur, Malaysia
Dr. Alptug AtilaDepartment of Analytical Chemistry,Faculty of Pharmacy, Ataturk University,Erzurum, Turkey
Dr. Gyanendra SinghSchool of Medicine, Louisiana StateUniversity Health Sciences Center NewOrleans, USA
CONSULTANT EDITORS
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Dr. Ajay Kumar GuptaUniversity Institute of Pharmacy,Chattrapati Sahu Ji MaharajUniversity, Kanpur, (U. P.), India
Dr. Padmini ShuklaAssistant Professor, UPRIMS & R, SafaiMedical College, Safai, Etawah (U.P.),India
Dr. Ramesh SabharwalMedical Officer, LNJP, Kurukshetra(Haryana), India
Dr. (Mrs.) Sanju NandaDepartment of Pharmaceutical Sciences M.D. University, Rohtak (Haryana), India
Prof. (Dr.) Amita VermaDepartment of Pharmaceutical Sciences,SHIATS, Allahabad (U.P.), India
Dr. Kuldeep SinghFaculty of Pharmacy, Integral University,Lucknow (U.P.), India
Dr. Prabodh ShuklaAssistant Professor, UPRIMS & R, SafaiMedical College, Safai, Etawah (U.P.),India
Dr. Shanti Bhushan MishraUnited Institute of Pharmacy, Allahabad(U.P.), India
Prof. (Dr.) Sanjay JainSmriti College of Pharmaceutical Sciences,Indore (M. P.), India
Prof. (Dr) B. GopalakrishnaR.R. College of Pharmacy, Bangalore(Karnataka), India
Prof. (Dr) A. R. KulkarniDepartment of Pharmacology, SETSCollege of Pharmacy, Dharwad(Karnataka), India
Prof. (Dr) Vimal JainInstitute of Pharmacy Nirma University,Ahmadabad (Gujarat), India
Prof. (Dr) A. K. GujjarInstitute of Pharmacy, Nirma University,Ahamdabad (Gujrat), India
Prof. (Dr.) Arun NandaDean, Faculty of Pharmaceutical SciencesM.D. University, Rohtak (Haryana), India
Prof. (Dr.) S. K. PrajapatiInstitute of Pharmacy, BundelkhandUniversity, Jhansi (U.P.), India
Dr. S. K. JainInstitute of Pharmacy, BundelkhandUniversity, Jhansi (U.P.), India
Prof. (Dr.) Shivesh JhaDepartment of Pharmaceutical Sciences,BIT, Mesra, Ranchi (Jharkhand), India
Dr. K. P. NamdevGuru Ghasidas Vishwavidhyalaya, Bilaspur(Chhattisgarh), India
Dr. (Mrs.) Savita VyasDepartment of Pharmacology, MGMMedical College, Indore (M. P.), India
Dr. Kumud UpadhyayaGIS Institute of Professional Studies,Dehradun, Uttarkhand, India
Dr. Abhishek MathurResearch Scientist (R&D) Sheetal LifeSciences, Dehradun (U.K), India
Prof. (Dr.) Ranjit SinghEx. Pro-Vice Chancellor, ShobhitUniversity, Meerut (U. P.), India
Dr. Ashutosh MishraPrincipal, A.N.D. College of Pharmacy,Babhnan, Gonda (U.P.), India
Dr. Asif HusainDepartment of Pharmaceutical ChemistryFaculty of Pharmacy, Hamdard University(Jamia Hamdard), New Delhi-110062
Dr. Vinod Kumar TiwariDepartment of Chemistry, Centre ofAdvanced Study , Banaras HinduUniversity Varanasi (U.P), India
Dr. N. K. TiwariThakral College of Technology, Orientalgroup of Institute, Bhopal (M. P.), India
Prof. (Dr.) Tejal A. MehtaInstitute of Pharmacy Nirma University,Ahamdabad (Gujrat), India
Dr. R. Irchhaiya Dr. Sushil K. Kashaw Dr. R. N. Gupta
EXECUTIVE EDITORS
ASSOCIATE EDITORS
EDITORIAL BOARD
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Department of Pharmacognosy, Institute ofPharmacy, Bundelkhand University, Jhansi(U.P), India
Department of Pharmaceutical Sciences,Dr. H S Gour Central University, Sagar (M.P.), India
Department of Pharmaceutical Sciences,BIT, Mesra, Ranchi (Jharkhand), India
Dr. Subha GangulyFaculty of Fishery Sciences, West BengalUniversity of Animal and Fishery Sciences,Kolkata, India
Dr. Amrish ChandraInstitute of Biomedical Education &Research, Manglayatan University,Beswan, Aligarh (U.P.), India
Dr. Sukhbir Lal KhokraInstitute of Pharmaceutical Sciences,Kurukshetra University, Kurukshetra(Haryana), India
Dr. Mahesh GuptaPrincipal Kota College of Pharmacy, Kota(Rajasthan), India
Dr. Himanshu PandeyDepartment of Pharmacy, SHIATS,Allahabad (U.P.), India
Dr. Nitesh KumarDepartment of Pharmacology & Toxicology,College Of Veterinary Science & AnimalHusbandry, Kuthulia, Rewa (M.P.), India
Mr. Rahul TanejaDepartment of Science &Technology,Government of Haryana ,Panchkula (H.R. ) ,India
Dr. Rohit GoyalDepartment of Pharmacology, ShooliniUniversity, Solan, (H.P.), India
Mr. Man SinghGovernment medical college, Allahabad(U.P.), India
Mr. Vivekanand KatareVivekanand College of Pharmacy Bhopal(M. P.), India
Dr. Prem Prakash SinghInstitute of Pharmacy, BundelkhandUniversity, Jhansi (U.P.), India
Dr. Anurag KhatkarDept. of Pharmaceutical SciencesM.D.University, Rohtak (Haryana), India
Dr. Lavkush DwivediInstitute of Biomedical SciencesBundelkhand University, Jhansi (U.P.),India
Dr. Ramesh SinghDepartment of Pharmacy RameshwaramInstitute of Tech. & Management Lucknow(U.P.), India
Ms. Preeti BohraResearch Scientist Ranbaxy LaboratoriesLtd. Mohali (Punjab), India
Mr. Sourabh KoseyDepartment of Pharmacy Practice, ISFCollege of Pharmacy, Moga, Punjab, India.
Dr. Aviral JainAdina Institute of Pharmaceutical SciencesSagar (M.P.), India
Dr. Pankaj ShuklaDepartment of Chemistry, Research lab,D.B.S. College Kanpur (U.P.), India
Mrs. Monika PandeyMahatma Ghandhi College ofPharmaceutical Sciences, Jaipur(Rajasthan), Iindia
Mr. Shobhit SinghInstitute of Pharmacy, BundelkhandUniversity Jhansi (U.P.), India
Mr. Devendra SinghInstitute of Pharmacy, BundelkhandUniversity, Jhansi (U.P.), India
Mr. V. K. SinghInstitute of Pharmacy, BundelkhandUniversity, Jhansi (U.P.), India
Mr. Piyush MantriAcropolis Institute ofTechnology & Research Indore (M.P.), India
Dr. Puspendra KumarAssistant Professor Department ofPharmaceutical Sciences Krishna Instituteof Engineering and Technology Ghaziabad,Uttar Pradesh, India
Mrs. Anuradha SinghInstitute of Pharmacy, Shree GanpatiInstitute of Technology, Ghaziabad (UP),India
Mr. Ramesh ChandAerosol Scientist and Chemist/SeniorResearch Associate Lovelace RespiratoryResearch Institute Albuquerque, NM, USA
Dr. Rupesh K. GautamAssociate Professor, Department ofPharmacology, ADINA Institute ofPharmaceutical Sciences, Sagar (M.P.),India
Dr. JagbirBPS Govt. Medical College for Women,Khanpur Kalan, District - Sonepat,Haryana, India
Mr. Rahul Deo YadavMotilal Nehru Medical College, Allahabad(U.P.), India
Dr. Neelesh MalviyaSmriti College of PharmaceuticalEducation, Indore (M.P.), India
Dr. Saurabh SatijaAssistant Professor , School ofPharmaceutical Sciences, LovelyProfessional University, Punjab, India
Dr. Meenu MehtaAssistant Professor, School ofPharmaceutical Sciences, LovelyProfessional University, Punjab, India
Dr. Nilesh JainAssociate Professor, Sagar Institute ofResearch Technology & Sciences -Pharmacy, Bhopal (MP), India
PUBLICATION COMMITTEE MEMBERS
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Dr. Pragati SainiLucknow (U.P.), India
Dr. S. RajvaidyaBhopal (M.P.), India
Mr. Vimal YadavJaunpur (U.P.), India
Dr. Namrata SinghNagpur (M.H.), India
Mr. Ashish MishraKanpur (U.P.), India
Mr. Dherendra GoswamiJhansi (U.P.), India
Mr. Gautam VermaGonda (U.P.), India
Ms. Neha SharmaBhopal (M.P.), India
Mr. Aman SantoshLalitpur (U.P.), India
Mrs. Swati JainGhaziabad (U.P.), India
Mrs. Archana OjhaGhaziabad (U.P.), India
Mr. Sumit Kant SinhaLucknow(U.P.) , India
Mrs. Noopur PandeyVadodara (Gujrat), India
Mr. Dilip Kumar ChanchalJhansi, (U.P.), India
Mr. Rohit Kumar BijauliyaJhansi, (U.P.), India
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Conference proceedings of the InternationalConference on Wellness will be published inIJPSR. The conference which will be held on8th-10th November 2019 at Chhatrapati ShahuJi Maharaj University, Kanpur (UP) India.
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Souvenir and Scientific abstract of 24th Annual NationalConvention of Association of Pharmaceutical Teachersof India (APTICON 2019), organized by Faculty ofPharmacy, DIT University, Dehradun, Uttarakhand inassociation with APTI Uttarakhand State Branch isavailable online.
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Search This Site...ISSN (Online): 0975-8232,ISSN (Print): 2320-5148
HomeHome About UsAbout Us Contact UsContact Us Search ArticlesSearch Articles
INTERNATIONAL JOURNAL OF PHARMACEUTICAL SCIENCES AND RESEARCHA Web of Science - ESCI Indexed Journal
Projected Impact Factor (2018): 0.83 , CiteScore (2017): 0.27
Five-Year Projected Impact Factor: 1.81
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HomeHome Volume 6 (2015) - Issue 3, March
Title Views PDF Cited
1.
Bibhas Kar and S. SivamaniCentre for Genetic Studies & Research, The Madras Medical Mission,Chennai-600037,TN, India.DOI: http://dx.doi.org/10.13040/IJPSR.0975-8232.6(3).940-50
940-950
Abstract HTML Full Text PDF Citation
APOPTOSIS: BASIC CONCEPTS, MECHANISMS AND CLINICAL IMPLICATIONS
Apoptosis, a well choreographed gene-directed cellular destruction, plays an important role in a variety ofbiological events, including morphogenesis, homeostatic maintenance of various tissues and removal of harmfulcells while dysregulated apoptosis has been implicated in a variety of disease such as cancer, systemic andorgan specific autoimmune disease and neurodegenerative disorders. Apoptos...
4106 1577 5
2.
Jyoti Mundlia , Mukesh Kumar and AmardeepAssistant Professor College of Pharmacy , PGIMS, Rohtak, Haryana, IndiaDOI: http://dx.doi.org/10.13040/IJPSR.0975-8232.6(3).951-60
951-960
Abstract HTML Full Text PDF Citation
NASAL DRUG DELIVERY- AN OVERVIEW
Over the past few decades the nasal route has gained widespread interest as a promising and an alternativeroute to oral and parenteral drug delivery. The nasal mucosa being highly vascularized and permeable provides arapid onset of therapeutic action. It is a convenient, compliant and needleless mode of drug delivery suitable forthe treatment of both acute and chronic diseases. In addition, nas...
4510 1351 9
3.
Anupam Kumar Sachan* and Ankita GuptaDayanand Dinanath College, Institute of Pharmacy, Kanpur, Uttar Pradesh,IndiaDOI: http://dx.doi.org/10.13040/IJPSR.0975-8232.6(3).961-70
961-970
Abstract HTML Full Text PDF Citation
A REVIEW ON NANOTIZED HERBAL DRUGS
A budding interest in Nanopharmaceuticals had generated a number of advancements throughout recent yearswith a focus on engineering novel applications. Nanophytomedicines are prepared from active phytoconstituentsor standardized extracts. The world market for nanomedicine is estimated to reach $130.9 billion by the fiscalyear 2016. Liposome nanoparticle (NP) with entrapped doxorubicin was repor...
5078 3094 12
4. MEDICINAL SIGNIFICANCE OF LOVASTATIN 4299 1425 2
Archive Most Popular Most Cited
Volume 6 (2015) - Issue 3, March
REVIEW ARTICLES
5/5/2020 Current Issues | INTERNATIONAL JOURNAL OF PHARMACEUTICAL SCIENCES AND RESEARCH
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9.
Marwa S. Elazazy *Analytical Chemistry Department, Faculty of Pharmacy, Zagazig University,Zagazig, EgyptDOI: http://dx.doi.org/ 10.13040/IJPSR.0975-8232.6(3).1022-32
1022-1032
Abstract HTML Full Text PDF Citation
INTERACTION OF TETRACYCLINE HYDROCHLORIDE WITH IRON: KINETIC SPECTROPHOTOMETRICAND CONDUCTOMETRIC INVESTIGATIONS
The interaction of the antibiotic, tetracycline hydrochloride (TC.HCl) has been investigated employing threesimple spectrophotometric and conductometric methods. In methods (A and B), a kinetic spectrophotometricprocedure based on the oxidation of (TC.HCl) by Fe3+ ion in the presence of 1, 10 - Phenanthroline (o-phen) (A)and 2, 2′- Bipyridyl (bipy) (B) was developed. The formation of the ferr...
2760 895 0
10.
R Lattanzi , C Congiu , V Onnis , A Deplano , S Salvadori , V Marconi , DMaftei , A Francioso , C Ambrosio , I Casella ,T Costa , G Caltabiano , M TMatsoukas , G Balboni and L Negri *Department of Physiology and Pharmacology ,Sapienza University of Rome,Rome, ItalyDOI: http://dx.doi.org/10.13040/IJPSR.0975-8232.6(3).1033-42
1033-1042
Abstract HTML Full Text PDF Citation
HALOGENATED TRIAZINEDIONES BEHAVE AS ANTAGONISTS OF PKR1: IN VITRO AND IN VIVOPHARMACOLOGICAL CHARACTERIZATION
Different prokineticin receptor antagonists, based on the triazinedione scaffold, were synthesized by a newefficient method. Here we demonstrated that 5-benzyltriazinediones substituted in position para of the benzylgroup with halogens provide compounds endowed with interesting selectivity for the Prokineticin receptor 1(PKR1). BRET technology indicates that such substitution results in increas...
2217 770 1
11.
M. A. A. Mamun , A. Rahman , S.H. Belal , M. A. Islam , M. E. H. Sarker , M. S.I. Arman , A. E. Ekram and K.M.F. Hoque *Protein Science Lab , Department of Genetic Engineering and Biotechnology,University of Rajshahi, Rajshahi , BangladeshDOI: http://dx.doi.org/10.13040/IJPSR.0975-8232.6(3).1043-48
1043-1048
Abstract HTML Full Text PDF Citation
HISTOLOGICAL STUDY OF THE EFFECT OF MALATHION ON LIVER AND KIDNEY TISSUES OF MICEMODEL
Malathion is a widely used organophosphorous pesticide that a large number of populations are undesirablyexposing themselves to severe health risk due to taking up the contaminated foods, water and vegetablescontaining malathion. The present study was carried out through histopathologic test to evaluate the extent ofdamage caused by malathion in the liver and kidney tissues of mice. Twenty five...
4188 1142 3
12.
Hamong Suharsono , Sumarno Reto Prawiro *, I Nyoman Mantik Hamong and MadeAgus HendrayanaLaboratory of Microbiology, Medical Faculty of Brawijaya University Malang,IndonesiaDOI: http://dx.doi.org/10.13040/IJPSR.0975-8232.6(3).1049-53
1049-105
Abstract HTML Full Text PDF Citation
IN – VIVO CONFIRMATION OF A 49.6 KDA PROTEIN PILI OF HELICOBACTER PYLORI TO PREVENTDESTRUCTION OF GASTRIC CELLS AGAINST LIVE HOMOLOGOUS BACTERIA IN MICE
Peptic ulcers is one of the major gastrointestinal disorder in human being that generally associated with theinfection of Helicobacter pylori, a gram-negative microaerophilic bacterium in stomach. It is also linked to thedevelopment of the stomach cancer. The aim of this study was to investigate the in vivo properties of the sub-unit pili proteins with molecular weight of about 49,6 kDa in m...
2157 740 1
13.
F. C. Saputri *, A. Mun’im , D. Lukmanto, S. N. Aisyah and J. S. RinandyPharmacology Laboratory, Faculty of Pharmacy, University of Indonesia,Kampus UI Depok-West Java , IndonesiaDOI: http://dx.doi.org/10.13040/IJPSR.0975-8232.6(3).1054-59
1054-1059
INHIBITION OF ANGIOTENSIN CONVERTING ENZYME (ACE) ACTIVITY BY SOME INDONESIA EDIBLEPLANTS
Antihypertensive properties of plant can be evaluated by in vitro method on inhibition of Angiotensin ConvertingEnzyme (ACE) activity. In this research, we investigate the inhibitory effect of several common edible plants onblocking ACE activity. ACE activity was evaluated by using N-hippuryl-L-histidyl-L-leucine (HHL) as substrate andthe inhibitory effect of extracts were determined based on t...
3829 1518 18
Suharsono et al., IJPSR, 2015; Vol. 6(3): 1049-1053. E-ISSN: 0975-8232; P-ISSN: 2320-5148
International Journal of Pharmaceutical Sciences and Research 1049
IJPSR (2015), Vol. 6, Issue 3 (Research Article)
Received on 21 July, 2014; received in revised form, 29 September, 2014; accepted, 01 December, 2014; published 01 March, 2015
IN - VIVO CONFIRMATION OF A 49.6 KDA PROTEIN PILI OF HELICOBACTER PYLORI TO
PREVENT DESTRUCTION OF GASTRIC CELLS AGAINST LIVE HOMOLOGOUS
BACTERIA IN MICE
Hamong Suharsono 1, Sumarno Reto Prawiro
*2, I Nyoman Mantik
3 and Made Agus Hendrayana
4
Laboratory of Biochemistry 1, Laboratory of Virology Veterinary
3, Laboratory of Microbiology
4, Faculty
of Udayana University Denpasar 80232, Bali-Indonesia
Laboratory of Microbiology 2, Medical Faculty of Brawijaya University Malang, Indonesia
ABSTRACT: Peptic ulcers is one of the major gastrointestinal disorder in human being
that generally associated with the infection of Helicobacter pylori, a gram-negative
microaerophilic bacterium in stomach. It is also linked to the development of the stomach
cancer. The aim of this study was to investigate the in vivo properties of the sub-unit pili
proteins with molecular weight of about 49,6 kDa in mice. The bacterium was firstly
cultured on the plate of TSA-B (Trypticase Soy Agar with 5% Sheep Blood) to prepare
the protein of interes using bacterial cutter and SDS-PAGE. The purified protein was
used for a vaccine emulsified with commercial cholera toxin and give orally. The
immunized mice showed a significant protection against challenge with live H. pylori
cells. In contrast, animals that received the 49, 6 kDa protein without adjuvant as well as
the negative control with PBS failed to inhibit adherence of the bacteria, as indicated by a
severe damages of gastric tissues. This study has indicated that the sub-unit pili proteins
trigered the release of protective antibodies againts the microorganism if combined with
cholere toxin adjuvant. Further study is required to investigate the biological functions of
this protein as a vaccine candidate for protecting the infection by this microorganism in
causing gastric ulcers.
INTRODUCTION: During the last decade it has
been established that the presence of microbes in
the stomach has been associated with gradual
increase of gastric cancer. The perception of the
bactericidal activity of stomach acid to hamper the
ability of bacteria to cause this condition has been
arguable. A number of investigators have
demonstrated that Helicobacter pyloriwas
considered as a major cause of gastric cancer 1, 2
.
The presence of the bacteria in the human stomach
coincided with a variety of gastric disorders such as
peptic ulcer, gastritis chronic and gastric carcinoma
even gastric lymphoma 3.
QUICK RESPONSE CODE
DOI: 10.13040/IJPSR.0975-8232.6(3).1049-53
Article can be accessed online on: www.ijpsr.com
DOI link: http://dx.doi.org/10.13040/IJPSR.0975-8232.6(3).1049-53
However, morbidity may varied in different some
individuals associated with acquired influences that
stimulate defenses of host against the infection4.
The ideal regimen to treat this infection has been
difficult primarily due to human habits including
smoking, overweigh, poor compliance 5, and re-
infections associated with antibiotic resistance 6.
However, eradication of the bacteria using various
therapeutic schemes could potentially prevent
gastric cancer 7. It has been demonstrated that H.
pylori colonizes the epithelial surface of the gastric
mucosa by forming a specific adhesion mechanism 8, with the involvement of immunogenic proteins
such as outer membrane proteins and sub-unit
proteins: Cag A, Vac A, adhesion A and Oip A 9
.
With this process, the microorganism must leave
the mucus membrane and adhere to the underlying
epithelium 10
. However, the precise mechanisms
Keywords:
Pili 49, 6 kDa, H. pylori,
cholera toxin, in vivo.
Correspondence to Author:
Sumarno Reto Prawiro
Laboratory of Microbiology, Medical
Faculty of Brawijaya University
Malang, Indonesia.
E-mail: [email protected]
Suharsono et al., IJPSR, 2015; Vol. 6(3): 1049-1053. E-ISSN: 0975-8232; P-ISSN: 2320-5148
International Journal of Pharmaceutical Sciences and Research 1050
underlying H. pylori adhesion have yet massively
to be identified.
It has been established that a sub-unit protein of H.
pylori with a molecular mass of about 49.6 kDa has
a pathogenic effect in causing peptic ulcer. A very
recent study has demonstrated that this protein was
found to be adherence on mice gastric epithelia
cells in vitro. Moreover, the attachment of intact H.
pylori cells on the purified mice gastric epithelial
cells could be protected by the presence of
polyclonal antibodies produced against the
homologous protein, indicating the protein was
dominant and immunogenic 11
.
However, the in vivo studies to demonstrate the
biological activities of the protein in causing
pathological consequences in the stomach are very
limited. The purpose of this study was to confirm
the in vivo pathogenesis of the 49.6 kDa protein of
H. Pylori in causing gastric epithelial cells damages
in mice. Furthermore the in vivo protection of the
epithelial cells by providing local vaccine
contained the pili protein against intact
Helicobacter pylori was also evaluated.
MATERIALSAND METHODS:
H. pylori isolate and cultivation:
A stock sample of H. pylori strain was kindly
provided by Biomedical Research Unit West Nusa
Tenggara Provincial Hospital. The bacterium was
originally isolated from patient with gastritis and
duodenum ulcer, and then re-cultured using media
Trypticase Soy Agar (TSA) and Trypticase Soy
Broth (TSB) supplemented with 10% sheep blood,
Dent supplement and Isovitalex and incubated at
37oC on microaerophilic atmosphere.
Subsequently, the bacteria were transferred into 10
ml sterile tubes containing about 106 cells/ ml and
kept for not more than1 hr at 5oC until used.
Isolation of H. pylori pili:
Isolation of H. pylori pili was performed by the
method 12
with a slight modification. Bacteria pili
was cut by using pili bacterial cutter which was
carried out for 30 sec at speed 5000 rpm while the
second to five cutting used the same speed.
Subsequently, the isolation of pili fraction by
centrifugation of cutting result was done at 12000
rpm at 4oC. The supernatant containing the
bacterial pili were stored at 4oC, a sample of it was
checked under an electron-microscope for
confirmation.
Isolation of H. pylori sub unit pili 49, 6 kDa
protein: Research methods
12 with slight modification. The
supernatant containing pili was done
electrophoretically by SDS-PAGE based on the
method 13
. The product of electrophoresis in the
form of gel was cut straight at a molecular weight
of about 49, 6 kDa. The gel pieces were then sliced
and inserted into the dialysis membrane by using
electrophoresis running buffer fluid. Subsequently,
the desired protein was electroeluted by placing the
membrane horizontally in the negative electrode
with current 20 mA for 15 minutes.
The dialysis was performed on the product of
electroellusion with PBS pH 7.4 buffer fluide as
much as 2 liters during 2 x 24 hours. Dialysis fluid
was changed three times, and dialysis fluid in
membrane dialysis as a result of electrophoellusion
of SDS-PAGE band was collected. Total protein
was measured using a method derived DC Protein
Assay (Bio-rad), suspended to a concentration of
about 10 ng per ml and kept at -20oC until used.
Confirmation of the sub unit pili 49, 6 kDa
protein that inhibits colonization of H. pylori in
gastic mucosa of mice:
Three groups of experimental mice consisting 4
mice in each group designed as group A, B and C
were used in this study. Mice in group A were
orally immunized with 0.5 ml of immunogen
contained 200 µg of sub unit pili 49,6 kDa protein
emulsified with 10 µg cholera toxin sub unit B, as
recommended by the factory (Sigma, USA). This
was repeated 3 times in 1 week intervals.
In contrast, animals in group B and C were only
orally given cholera toxin and PBS respectively. At
fourth week of experiment, all animals in the three
groups were challenged orally with 0.5 ml of live
H. pylori at a concentration of about 106 cells/ml,
the clinical signs were observed daily. The animals
were kept for one week before being killed for
gastro-intestinal sample collections. The samples
were then process using H&E staining as published
previously.
Suharsono et al., IJPSR, 2015; Vol. 6(3): 1049-1053. E-ISSN: 0975-8232; P-ISSN: 2320-5148
International Journal of Pharmaceutical Sciences and Research 1051
RESULTS:
Confirmation that H. pylori pili was isolated:
To confirm if pili of H. pylori could be isolated as
desired was checked by observing a sample of it
under electron-microscope, before being used for
further study. This study found that the pili was
isolated as expected, indicated by a uniform white
sediments without any cellular morphology (Fig.
1A), compared to the whole cells indicated by
intact cells connected with pili (Fig. B)
A B FIGURE 1: MORPHOLOGICAL FEATURE OF
ISOLATED PILI (A) AND WHOLE CELLS (B) OF H.
PYLORI UNDER ELECTRON MICROSCOPE
EXAMINATIONS, THE MAGNIFICATIONS ARE
NOTED.
Isolation of H. pylori subunit pili 49, 6 kDa
protein:
A sample of the isolated pili was subjected into
SDS-PAGE to localized the position of the sub unit
pili 49, 6 kDa protein, before being cut and
purified. As it was published previously, the
position the protein of interest was quite clear with
molecular weight of about 49, 6 kDa, suggesting
the desired protein could precisely be localized
(Fig.2 A). Furthermore, this protein was assumed
to be immune-dominant and immunogenic,
indicating by a strongest protein band compared to
other proteins in Western blot analysis (Fig. 2 B).
This protein band was then purified for the
immunization of mice, although product of the
purified protein is not shown.
In Vivo confirmation of the sub unit pili 49, 6
kDa protein to prevent colonization of H. pylori
in gastic mucosa of mice:
The immunized mice with the sub unit pili 49, 6
kDa H. pylori demonstrated a protective reaction
against live infection of H. pylori. There were no
significant damages of gastrical tissues found after
one week incubation in vivo (Fig. 3a). In contrast, a
severe lost of epithelial cells was observed in
animal without the presence of cholera toxin
adjuvant in the inoculums given (Fig. 3b) and in
negative control with PBS (Fig. 3c). No lesions
were observed in normal tissues (3d).
A B
FIGURE 2: THE PRECISE LOCATION OF THE SUB
UNIT PILI 49,6 kDa PROTEIN OF H. PYLORI WAS
CONFIRMED BY USING SDS-PAGE (A) AND
WESTERN BLOT ANALYSIS (B). THE POSITION OF
THE PROTEIN IS INDICATED (ARROWED).
FIG. 3A FIG.3B FIG. 3C FIG. 3D
FIGURE 3: DEMONSTRATION INFECTION OF
INTACT CELL OF H. PYLORI IN VARIOUS ANIMAL
TREATMENTS.
A significant protection of tissue in animals
immunized with sub unit pili 49,6 kDa (3a); a
serious tissue damages observed in non-immunized
animals (3b and 3c) after challanged with live
bacterial cells; and a normal feature of gastrical
tissues (3d).
DISCUSSION: The initial step of colonization
process by H. pylori is their ability to adhere the
mucosal surface of gastric epithelial cells. The
adhesion of H. pylori to mucus constituents in the
human stomach is a predisposition site for the
attachment of this unique niche, which has become
the major habitat of the microorganism 14
. In other
bacteria, it was reported that a sub unit pili protein
of Shigella dysentriae and Salmonella typhi with
molecular weight of 49, 8 kDA and 48 kDa
Suharsono et al., IJPSR, 2015; Vol. 6(3): 1049-1053. E-ISSN: 0975-8232; P-ISSN: 2320-5148
International Journal of Pharmaceutical Sciences and Research 1052
respectively were hemaglutinin with adhesion
properties 15
. With H. pylori it selves, it has just
recently published that a sub unit pili 49, 6 kDa
protein of this microorganism was found to be
immunogenic and immune dominant in vitro 11
. For
this, we reasoned that this protein may be a
potential candidate for protection against natural H.
pylori infections.
In this study, the in vivo biological properties of the
sub unit pili 49,6 kDa protein of H. pylori were
investigated. It was previously published that this
protein could be purified for the in vitro studies
which showed its immunogenicity 11
. In animal
models, application of this protein mixed with a
commercial cholera toxin adjuvant, revealed a
significant inhibition against live H. pylori
challenge in the mucosal gastric. It was reported
that H. pylori possess a a type IV secretion system
encoded by the cag pathogenicity island, the
effector protein CagA, the vacuolating cytoxin
(VacA) and others that responsible in causing
pathological effect in H. pylori infections 16
.
This feature was not experienced in this study,
suggesting that the sub unit pili 49, 6 kDa protein
may has a crossed-protected reactivity to the toxin
in vivo. However, compared to the normal samples
prepared from normal animals, the degree of
protection was less complete.
This may be due to vaccination regime used, in
which the dose of antigen and adjuvant need to be
further optimized. In contrast, the introduction of
the protein alone, without the adjuvant failed to
protect gastric mucosa, suggesting that protein was
immunogenic as reported previously and the
adjuvant play important role in providing a good
protection. With this regard, inhibition of H. pylori
adhesion to human gastric mucus was proved by
giving a high-molecular weight constituent of
cranberry juice 10
. However, it seemed likely that
the inhibition by using the juice may be due to
physical protection, rather than biological reactions
as it was reported in the current study.
In line with our previous data that, application of
the isolated IgG on the pre-treated mice gastric
epithelial cells with the purified sub unit pili 49, 6
kDa proteins, repealed an inhibition of H. pylori
cells to adhere the cells. This further suggested that
the sub unit pili 49, 6 kDa proteins had a specific
adhesion molecules to bind the target gastric cells
of mice. The inability of the bacteria to attach the
gastric epithelial cells particularly when a high titer
of IgG was added, confirming the sub unit pili 49,6
kDa protein was a functional protein that may
associated with the pathogenesis of the bacteria.
The in vivo study reported here has confirmed that
the protein could provide a protection against
natural infection as demonstrated by histological
observations.
However, the protection was specific only against
the H. pylori cells, but not against cholera toxin
(data not shown).
This result suggested that the both the sub unit pili
49,6 kDa protein of the bacteria and cholera toxin
as an adjuvant had synergetic actions in protecting
the gastric tissues in mice against live challenge of
H. pylori, and therefore it may be necessary to use
this protein as a vaccine candidate for the
prevention against H. pylori infections.
CONCLUSION: The in vivo study reported here
has further confirmed the in vitro characteristic of
the 49, 6 kDa sub unit pili protein the H. pylori that
was previously published. The current result has
suggested that the protein was also found
immunogenic, causing significant damages to
gastric tissues of infected mice. However, the
corporation this protein with a commercial cholera
toxin adjuvant and given orally, indicated that the
49, 6 kDa sub unit pili protein the H. pylori was an
useful antigen associated with protection against
this microorganism when emulsified with the of
cholera toxin. Further study is required to
investigate the biological functions of this protein
as a vaccine candidate for protecting the infection
by this microorganism in causing gastric ulcers.
ACKNOWLEDGMENT: The authors express
many thanks for giving fund and opportunity by
our Ministry of Health Indonesia in the format of
IPTEDOK for the study.
CONFLICT OF INTEREST: The authors report
no conflict of interest.
Suharsono et al., IJPSR, 2015; Vol. 6(3): 1049-1053. E-ISSN: 0975-8232; P-ISSN: 2320-5148
International Journal of Pharmaceutical Sciences and Research 1053
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How to cite this article:
Suharsono H, Prawiro SR, Mantik IN and Hendrayana MA: In - Vivo Confirmation of A 49.6 Kda Protein Pili of Helicobacter Pylori to
Prevent Destruction of Gastric Cells Against Live Homologous Bacteria In Mice. Int J Pharm Sci Res 2015; 6(3): 1049-53.doi:
10.13040/IJPSR.0975-8232.6(3).1049-53.