INTERNATIONAL JOURNAL OF SCIENTIFIC & INNOVATIVE RESEARCH (A Bi-annual Official Journal of Sky Institute, a constituent Institution of BALA JI FOUNDATION established under Societies Registration Act 1860). The International Journal of Scientific & Innovative Research (IJSIR) is a bi-annual published on line/ offline journal which publishes innovative research papers, reviews, mini reviews

1. VOLUME – 4, ISSUE – 2 (JULY-DECEMBER) 2016

2. Editorial
I am happy to convey that second issue of third volume of "International Journal of Scientific
and Innovative Research (IJSIR)", a bi-annual journal has been published by Sky Institute,
Lucknow in an effort to promote multidisciplinary scientific and innovative research of
societal benefit. This journal covers all branches of science, technology, engineering, health,
agriculture and management. Research articles in the field of education are also encouraged in
order to promote educational technology aiming at improvement in present educational
system. As research and development (R & D) has been playing a significant role in overall
development of society, continuous multidisciplinary innovative research in science and
technology is needed to address the challenges in context to changing environmental
conditions in the present era of gradual increase in industrial and technological advancement
at global level. Efforts should be made to develop eco-friendly technologies in order to
providesolutionsfordevelopingsocially,economicallyandculturallysustainablesociety.
The present issue of International Journal of Scientific and Innovative Research (IJSIR)
contains 12 research papers I articles covering different areas of science and technology. All
these papers are well written and informative in content. I express my sincere thanks and
gratefulness to Mr. Mohit Bajpai, Chairman, Sky Institute, Lucknow (U.P.), India for his
support in publishing it. I express my thanks to members of Committee for Editorial
Assistance Dr. B.C.Tripathi, Dr. Pankaj Verma, Shri Sanjay Pandey, Shri Sanjay Dixit and
Mr. Shamshul Hasan Khan for their hard work and devotion in giving the final shape to the
journal. I am thankful to all faculty members, scientists and research scholars of different
universities, research organizations and technical institutions for contributing their research
articles for publication in the present issue of the journal. The help provided by faculty
members and supporting staff of Sky Institute in publishing the present volume of the journal
is also acknowledged. I hope scientists, academicians and young researchers will be greatly
benefitedbythispublicationfortheirresearchwork.
I request humbly to the readers and contributors of our journal to continue encouraging us for
regular publication of the journal. Any suggestion and comment for the improvement in the
qualityofthejournalarealwayswelcome.
Dr. B. R. Pandey
Editor-in-Chief

3. EDITOR-IN-CHIEF
Dr. B.R. Pandey
Director (Research)
Sky Institute, Kursi Road, Lucknow, U.P, India
Dean, Faculty of Science & Technology, Sai Nath University, Jharkhand, India
Former Joint Director, Council of Science & Technology, UP, Lucknow
(Department of Science and Technology, UP Government), India
Former Professor, International Institute of Herbal Medicine (IIHM), Lucknow, U.P., India
E-mail Id: editorijsir02@gmail.com, Mobile-: 9794849800
COMMITTEE FOR EDITORIALASSISTANCE
Dr. B.C.Tripathi
Assistant Prof., Deptt. of Education,
Rama P.G. College, Chinhat,
Lucknow, Uttar Pradesh
Dr. Pankaj Verma
Senior Research Fellow, Deptt. of
Oral & Maxillofacial Surgery, Faculty
of Dental Sciences, K.G. Medical
University, Lucknow, Uttar Pradesh
Shri Sanjay Pandey
Assistant Prof., National Institute
of Fashion Technology, Raebareli,
Uttar Pradesh
Shri Ashish Tiwari
Research Scholar, Sai Nath University,
Ranchi, Jharkhand
Shri Sanjay Dixit
Scientist, Sky Institute, Lucknow,
Uttar Pradesh
Shamshul Hasan Khan
Scientist, Sky Institute, Lucknow,
Uttar Pradesh
ADVISORY BOARD
Prof.(Dr.)S. P. Ojha
Former Vice Chancellor, CCS Meerut University, Meerut, Uttar Pradesh
Prof.(Dr.)V.K. Srivastava
Former Prof & Head, Deptt. of Community Medicine King George Medical
University, Lucknow.
Former Director, Integral Institute of Medical Sciences & Research, Integral
University, Lucknow
Former Vice -Chancellor,
Texila American University, Georgetown, Guyana, South America
Prof.(Dr.) M.I. Khan
Prof & Head, Deptt. of Mechanical Engg., Integral University, Lucknow, Uttar
Pradesh
Prof. (Dr.) S.K. Avasthi
Former Director, H.B.T.I., Kanpur, Uttar Pradesh
Prof.(Dr.) Amrika Singh
Prof & Head (Chemistry), Deptt. of Applied Sciences,
Institute of Engg. & Technology, Sitapur Road, Lucknow, Uttar Pradesh
Prof. (Dr.) U.N. Dwivedi
Prof & Ex- Head, Deptt of Biochemistry, Former Pro- Vice Chancellor, Former
Dean, Faculty of Science, University of Lucknow, Lucknow, U.P.
Prof. (Dr.) U.K. Misra
Head, Deptt. of Neurology, Ex Dean,
Sanjay Gandhi Post Graduate Institute of Medical Sciences, Lucknow, U.P.
Dr. A.K. Gupta
Former Deputy Director General,
Indian Council of Medical Research (ICMR), Ansari Nagar, New Delhi
Prof.(Dr.) V.K.Tondon
Former Prof & Head, Deptt. of Chemistry, Ex- Dean Faculty of Science,
University of Lucknow, Lucknow, Uttar Pradesh
Prof. (Dr.) Amod Kumar Tiwari,
Prof.- Director, Bhabha Institute of Engg.& Technology, Kanpur, U.P.
Prof.(Dr.) Chandra Dhar Dwivedi
Former Prof. & Chairman, Deptt. of Pharmaceutical Sciences, College of
Pharmacy, South Dakota State University, Borokings, South Dakota, USA
Prof. (Dr.) Vimal Kishore
Prof. & Chairman, Deptt. of Basic Pharmaceutical Sciences,
Xevier College of Pharmacy, University of Louisiana, 7325, Palmetto Street
New Orlens, Louisiana USA
Prof. (Dr.) S.P. Singh
Former Prof & Head, Deptt. of Pharmacology,
G. S. V. M. Medical College, Kanpur, Uttar Pradesh
Prof. (Dr.) R. L. Singh
Prof & Head, Department of Biochemistry & Coordinator Biotechnology
Program , Dr. R. M. L. University Faizabad, Uttar Pradesh
Dr. Sarita Verma
Head, Deptt. of Home Sci., Mahila P.G. College, Kanpur, Uttar Pradesh
Prof. (Dr.) S.K.Agarwal
Pro. & Ex-Head, Deptt. of Biochemistry, Lucknow University, Lucknow,
U.P.
Dr. Bharat Sah
Director,
National Institute of Fashion Technology, Raebareli, Uttar Pradesh
Prof. (Dr.)N.S. Verma
Prof., Deptt. of Physiology,
K. G. Medical University, Lucknow, Uttar Pradesh
Prof. (Dr.)A.K. Tripathi
Prof. & Head, Deptt. of Clinical Hematology & Medical Oncology,
K. G. Medical University, Lucknow, Uttar Pradesh
Prof.(Dr.)C.M. Pandey
Prof. & Head, Deptt. of Biostatistics & Health Informatics,
Sanjay Gandhi Post Graduate Institute of Medical Sciences, Lucknow,
Uttar Pradesh
Dr. Rupesh Chaturvedi
Associate Prof., School of Biotechnology,
Jawaharlal Nehru University, New Delhi, Former Asstt. Prof., Deptt. of
Pharmaceutical Sciences , College of Pharmacy, Vanderbilt University,
Tennessee, USA
Dr. S.Sinha
Asstt. Prof. Deptt. of Internal Medicine, CD University,
C. David Giffen School of Medi., University of California, Los Angeles, USA
Dr. K.Raman
Principal Scientist, Martek Biosciences Corporation, 6480 Dobbin Road,
Columbia, MD 21045, USA Dr. P.K.Agarwal
Editor –in – Chief, Natural Product Communication,
Natural Product Inc 7963, Anderson Park Lane West Terville, OH, USA
Dr. R.K.Singh,
Chief Scientist, Division of Toxicology, CSIR-Central Drug Research
Institute, Jankipuram Extension, Lucknow, Uttar Pradesh
Dr. Mohd. Tarique
Prof., Deptt of Physical Edu., Lucknow University, Lucknow, Uttar Pradesh
International Journal of Scientific and Innovative Research 2016; 4(2)
P‐ISSN 2347‐2189, E‐ ISSN 2347‐4971
www.ijsir.co.in

4. EDITORIAL BOARD
Prof.(Dr.) Y.B. Tripathi
Prof. & Head, Deptt. of Medicinal Chemistry,Institute of Medical Sciences, Banaras
Hindu University Varanasi, Uttar Pradesh
Prof.(Dr.) R.K. Singh
Prof. & Head , Deptt. of Biochemistry, Shri Guru Ram RaiInstitute of Medical & Health
Sciences, Dehradun, Uttarakhand & Former Prof. & Head, Department of Biochemistry,
K. G. Medical University, Lucknow, U.P.
Prof. (Dr.) R.S.Diwedi
Former Director, National Research Centre for Groundnut (NRCG) , ICAR, Junagarh,
Gujarat & Former Principal Scientist – Head, Deptt. of Plant Physiology, Indian Institute
of Sugarcane Research, Lucknow, Uttar Pradesh
Prof. (Dr.) Nuzhat Husain
Prof. & Head , Deptt of Pathology & Acting Director, R. M. L. Institute of Medical
Sciences, Lucknow,Uttar Pradesh
Prof. (Dr.) Amita Jain
Prof. Deptt. of Microbiology, K.G. Medical University, Lucknow, U.P.
Dr. Sudhir Mahrotra
Associate Prof., Deptt. of Biochemistry, Lucknow University, Lucknow, U.P.
Prof. (Dr.) Vibha Singh
Prof., Deptt. of Oral & Maxillofacial Surgery, Faculty of Dental Sciences,
K. G. Medical University, Lucknow, Uttar Pradesh
Prof. (Dr.) U.S. Pal
Prof. & Head, Deptt. of Oral & Maxillofacial Surgery, Faculty of Dental Sciences,
K. G. Medical University, Lucknow, Uttar Pradesh
Prof. (Dr. ) K.K. Pant
Prof. & Head, Deptt. of Pharmacology & Therapeutics,
K. G. Medical University, Lucknow, Uttar Pradesh
Dr. C.M.K.Tripathi
Former Deputy Director & Head, Division of Fermentation Technology, CSIR- Central
Drug Research Institute , Lucknow, Uttar Pradesh
Dr. R.D. Tripathi
Chief Scientist & ProfessorPlant Ecology & Environmental Science Division, Uttar
Pradesh CSIR-National Botanical Research Institute, Lucknow, U.P.
Prof.(Dr.) Ashwani K. Srivastav
Prof. & Head, Deptt. of Biosciences, Integral University,Lucknow,
Former Senior Scientist, Birbal Sbahani Institute Paleobotany, Lucknow, U.P.
Prof.(Dr.) L. Pandey
Prof. & Head , Postgraduate Deptt . of Physics,Former Dean, Faculty of Science, Rani
Durgawati University, Jabalpur, Madhya Pradesh, India
Prof .(Dr.) Bali Ram
Prof., Deptt. of Chemistry, Banaras Hindu University, Varanasi, Uttar Pradesh
Prof.(Dr.) J.P.N.Rai
Prof.& Head, Deptt. of Environmental Sciences, G.B. Pant University of Agr. &
Technology, Pant Nagar, Uttarakhand
Prof.(Dr. )R. S. Dubey
Prof. & Head, Deptt. of Biochemistry, Banaras Hindu University, Varanasi, U.P.
Prof. (Dr.) Omkar
Deptt. of Zoology, Lucknow University, Lucknow, Uttar Pradesh
Prof.(Dr.) Sudhir Kumar
Prof., Deptt. of Zoology, Lucknow University, Lucknow, Uttar Pradesh
Prof.(Dr.) Naveen Khare
Prof., Deptt. of Chemistry, Lucknow University, Lucknow, Uttar Pradesh
Prof.(Dr.) S. M. Natu
Prof., Deptt. of Pathalogy,K.G. Medical University, Lucknow, Uttar Pradesh
Dr. Kusum Lata Mishra,
In-charge, Coagulation Laboratory, Deptt. of Pathology,
K.G. Medical University, Lucknow, Uttar Pradesh
Prof.(Dr.)V.K. Sharma,
Prof., Deptt. of Chemistry, Lucknow University, Uttar Pradesh
Prof.(Dr.) R.K. Shukla
Prof., Deptt. of Physics, Lucknow University, Lucknow Uttar Pradesh
Prof.(Dr.)Anil Gaur
Prof., Deptt. of Biotechnology & Genetic Engg., G.B. Pant University of Agr. &
Technology, Pant Nagar, Uttarakhand
Dr. Mahesh Pal
Principal Scientist ,Phytochemistry Division, CSIR- National Botanical Research
Institute, Lucknow, Uttar Pradesh
Dr. Vinod Singh
Assoc. Prof. & Head, Deptt. of Microbiology, Baruktulla University, Bhopal,
Madhya Pradesh
Dr. K.K.Verma
Assoc. Prof., Deptt. of Physics & Electronics.Dr. R. M. L. Awadh University ,
Faizabad,Uttar Pradesh
Dr. Atul Gupta
Senior Scientist, CSIR- Central Institute of Medicinal & Aromatic Plants,
Lucknow, Uttar Pradesh
Dr. Saudan Singh,
Senior Principal Scientist, CSIR- Central Institute of Medicinal & Aromatic
Plants , Lucknow, Uttar Pradesh
Dr. S.K.Tiwari
Senior Principal Scientist ,CSIR- National Botanical Research Institute,
Lucknow, Uttar Pradesh
Dr. Shivani Pandey,
Asstt. Prof., Deptt. of Biochemistry,K.G.Medical University, Lucknow, U.P.
Dr. B.C. Yadav,
Lucknow Associate Prof. & Coordinator, Deptt. of Applied Physics, School for
Physical Sciences, Babasaheb Bhimrao Ambedkar University, Lucknow, U.P.
Dr. Anchal Srivastava,
Prof., Deptt of Physics, Lucknow University,Lucknow, Uttar Pradesh
Dr. Shalini Bariar
Associate Professor, Thakur Institute of Management Studies and
Research,, Mumbai, India
Dr.A.K.Pandey
Principal Scientist, National Bureau of Fish Genetic Resources,Lucknow, U.P.
Dr.S.K.Pandey
G.M. LML Factory, Kanpur Uttar Pradesh
Dr. Suneet Kumar Awasthi,
Asst. Prof, Deptt.of Physics J.P. University, Noida, Uttar Pradesh
Dr.G. N. Pandey
Asst. Prof, Deptt. of Physics Amity University, Noida ,Uttar Pradesh
Dr. Mukesh Verma
Asst. Prof., Deptt. of Physical Education, Dr. R.M.L. Avadh University,
Faizabad, Uttar Pradesh
Dr. Abhay Singh,
Head, Physical Education, Delhi Public School, Lucknow Uttar Pradesh
Dr. Santosh Gaur
Asst. Prof. Deptt. of Physical Education, Jawahar Lal Nehru P.G. College,
Barabanki, Uttar Pradesh
Dr.Sanjeev Kumar Jha
Senior Scientist, DEOACC Patna
Dr. Shivlok Singh
Scientist, DEOACC, Lucknow, Uttar Pradesh
Dr. Anurag Tripathi,
Asstt . Prof. , Deptt. of Electrical Engg., Institute of Engg. & Technology,
Sitapur Road, Lucknow, Uttar Pradesh
Prof. V.P.Sharma
Senior Principal Scientist, CSIR-Indian Institute of Toxicology Research,
Lucknow, Uttar Pradesh
Dr. Krishna Gopal
Former Deputy Director & Head , Aquatic Toxicology Division, CSIR- Indian
Institute of Toxicology Research, Lucknow, Uttar Pradesh
Dr. S.P. Shukla
Prof. , Deptt. of Civil Engg., Institute of Engg. & Technology, Sitapur Road ,
Lucknow, Uttar Pradesh
Dr. Ajay Mishra
Associate Prof. , Deptt. of Geology, Lucknow University, Lucknow , U. P.
Dr. Ashutosh Singh
Prof., Deptt. of Chemistry,Saket P.G. College, Ayodhya, Faizabad, U. P.
Dr. S.K. Singh
Principal, Gita College of Education , Nimbari, Panipat, Haryana
Shri Sudesh Bhat
Advisor (Education), Sky Institute, Lucknow, Uttar Pradesh
Dr. Krishna Gopal
Asst. Prof., Deptt. of English,Rama University, Kanpur, Uttar Pradesh
International Journal of Scientific and Innovative Research 2016; 4(2)
P‐ISSN 2347‐2189, E‐ ISSN 2347‐4971
www.ijsir.co.in

5. ABOUT EDITOR-IN- CHIEF : DR. B. R. PANDEY
Dr. B. R. Pandey is a well-known academician and scientist with brilliant academic career and research
accomplishments. He has done M.Sc. (organic chemistry) from Banaras Hindu University,Varanasi, India in
the year 1972. He has done PhD in Medicinal Chemistry under the guidance of world renowned Biochemist
& Medicinal Chemist, Professor S.S. Parmar, Professor of Medicinal Chemistry & Chemical Pharmacology,
Department of Pharmacology & Therapeutics, K. G. Medical College, Lucknow (Presently K. G. Medical
University), Faculty of Medicine, University of Lucknow, Lucknow, India in the year 1976. Dr. Pandey has
all throughout first class educational qualifications and his research interest covers medicinal chemistry,
biochemical pharmacology, neurochemistry, neuro-toxicology, environmental chemistry, herbal medicine
& natural products. He is having extensive research experience of more than 40 years and published several
research papers in peer reviewed journals of international repute. His research particularly on the studies of
central nervous system acting drugs and anti-inflammatory drugs and their biochemical mode of action
using animal models and enzymes such as monoamine oxidase, acetylcholine esterase, purine catabolizing
enzymes , proteolytic enzymes, membrane stabilizing enzymes, respiratory enzymes, microsomal enzymes
etc. has been well recognized as evidenced by his research publications. Further, his research on developing
herbal medicines has been found very useful in prevention and treatment of chronic diseases and other
refractory diseases for which modern system of medicine have no permanent cure. He has worked on the
position of Joint Director, Council of Science & Technology, U.P., Lucknow, Department of Science &
Technology, Uttar Pradesh Government, India from the year 1979 to 2011, where he successfully executed
several R & D projects in various disciplines of Science &Technology including chemical & pharmaceutical
sciences, medical sciences, biological sciences, environmental sciences etc. During his tenure as Joint
Director, he has been instrumental in launching and implementing important schemes: Young Scientists
Scheme,Young Scientist Visiting Fellowship Scheme, Establishment of Centre of Excellence- Encephalitis
Research Centre of Excellence in Sanjay Gandhi Post Graduate Institute of Medical Sciences ( SGPGIMS),
Lucknow , U. P. India ; Centre of Excellence in Materials Science ( nano materials) in Z. H. College of Engg.
& Technology,Aligarh Muslim University,Aligarh, U.P. India, Establishment of Patent Information Centre
in the premises of Council of Science & Technology , U.P. He has also worked on the post of Secretary ( as
additional charge ) , Council of Science & Technology, U.P. several times and functioned asAdministrative
Head of the Organization. Prior to taking over the position of Joint Director, Council of Science &
Technology, U.P. in the year 1979, he has worked as Junior Research Fellow/ Senior Research Fellow
(Council of Scientific & Industrial Research, New Delhi ), Assistant Research Officer ( Jawaharlal Nehru
Laboratory of Molecular Biology) at Department of Pharmacology & Therapeutics, K.G. Medical College
(presently K. G. Medical University), Faculty of Medicine, University of Lucknow, Lucknow, India from
the year 1972 to 1979 and involved in multidisciplinary biomedical research leading to drug development .
He has worked as Visiting Scientist / Faculty in the Department of Physiology, School of Medicine,
University of North Dakota, Grand Forks, North Dakota, USA and also visited scientific institutions in
Sweden, U.K. and U.S.A. under Training Program on Capacity Building in Environmental Research
Management (World Bank Funding Project). After his superannuation in the year 2011, he has been
associated with International Institute of Herbal Medicine (IIHM), Lucknow, India as Professor and is
presently associated with Sky Institute, Lucknow, India as Director (Research) and Dean, Faculty of Science
& Technology, Sai Nath University, Jharkhand, India and involved in programs related to higher education
andresearchof scientific&technologicalfields. Hehas organizedseveralnationalandinternational
International Journal of Scientific and Innovative Research 2016; 4(2)
P‐ISSN 2347‐2189, E‐ ISSN 2347‐4971
www.ijsir.co.in

6. conferences. He has actively participated in national and international conferences, symposia and
workshops and presented research papers and chaired scientific / technical sessions. He is life member and
fellow of many scientific societies such as National Academy of Sciences India, Society of Toxicology of
India, Indian Academy of Neurosciences, Bioved Research Society India, International Society for Herbal
Medicine (ISHM), Society of Biological Sciences and Rural Development, India. He has been member of
several scientific expert committees/ advisory committees to evaluate scientific research proposals. Dr.
Pandey has been actively associated with various universities and institutions in India as examiner for
conducting graduate, post graduate and doctoral level examinations in disciplines like chemical sciences,
pharmaceutical sciences, biochemical sciences, biotechnology and allied areas and member of Board of
Studies for the academic development in the department. He has been approved research supervisor for
guiding research in chemistry, biotechnology and related areas from various universities of India leading to
PhD Degree. In view of his vast research and administrative experience and broad R & D vision, Dr. Pandey
has been associated with International Journal of Scientific & Innovative Research (IJSIR) as Editor-in-
Chief.
International Journal of Scientific and Innovative Research 2016; 4(2)
P‐ISSN 2347‐2189, E‐ ISSN 2347‐4971
www.ijsir.co.in

7. FROM THE DESK OF CHAIRMAN, SKY INSTITUTE
It is my privilege to state that I have great desire to contribute to the development of our
country and to bring about social transformation through education, higher learning and
research. This inner feeling prompted me to establish Sky Institute in Lucknow (Uttar
Pradesh), the city known for its rich cultural heritage and vibrant academic institutions
of higher learning. Sky Institute, since its inception in the year 2006, has been
functioning to impart various educational and training courses with a vision to improving lives through
education, research and innovation.The institute provides a professional learning environment that acts as a
catalyst, for the exponential growth of student as well as extracurricular abilities. It conducts regular courses
at the level of graduate and post graduate followed by research courses leading to M Phil and PhD in all
subjectsinassociationwithuniversities.
I feel great pleasure to highlight that Sky Institute has started to publish a bi-annual journal “International
Journal of Scientific and Innovative Research ( IJSIR ) which encourages to publish research articles in all
branches of science, technology, engineering, health, agriculture and management. Research articles in the
field of education are also considered in order to improve educational standard in educational institutions
with innovative technologies. First volume of the journal has been successfully published.The present issue
of second volume of the journal contains useful and informative research articles which may be interesting
to readers and educational and research organizations. The association of eminent faculty and scientists of
reputedorganizationswithourjournalishighlyappreciable.
I call upon all the students who are willing to join various programs/courses being run at Sky Institute in
association with selected universities, to strive hard to gain knowledge, transform it into skills with right
attitudeandinculcatethehabitof learning,whichwilldrivethemtoselfdirectedlearning.
My bestwishes toalltheaspiringstudents.
Mohit Bajpai
Chairman
International Journal of Scientific and Innovative Research 2016; 4(2)
P‐ISSN 2347‐2189, E‐ ISSN 2347‐4971
www.ijsir.co.in

8. CONTENTS
International Journal of Scientific and Innovative Research 2016; 4(2)
P‐ISSN 2347‐2189, E‐ ISSN 2347‐4971
www.ijsir.co.in
GEOMETRICALAND CONFORMATIONAL STERIC AFFINITY INTERACTION 1
STUDIES OF ANTICANCER DRUGS WITH HEXANUCLEOTIDE SEQUENCES OF
DNA BY FLUORESCENCE AND ABSORPTION SPECTROPHOTOMETRY.
Sulabh Kharbanda
PREVALENCE AND STUDY OF DIFFERENT STAGES OF CHRONIC KIDNEY DISEASES (CKD) 9
IN PATIENTS.
Shailza Verma, Dheeraj Upadhyay, Rahul Singh, Neha Pandey, Saurabh Singh,Vishwajeet Pandey, Sulabh Kharbanda
HONEY- GOLDEN LIQUID FOR HEALTH 16
Vibha Singh
PHENYLHYDRAZINE INDUCED HAEMATOTOXICITY AND IT'S RECOVERY BY FUMARIA 22
INDICA PLANT EXTRACT IN CHARLES FOSTER RATS
Raj Kumar, Vivek Kumar Mishra, Anil K.Meena, Pallavi Singh, R.L.Singh and R .K. Singh
IN VIVO ASSESSMENT OF ANTI-GENOTOXIC POTENTIAL OF 32
ASPARAGUS ADSCENDENS
Sakshi Jaiswal, Anil K. Meena, Pallavi Singh, R.L. Singh and & R.K. Singh
BENZENE INDUCED HAEMATOTOXICITY AND IT'S AMELIORATION BY 43
QUERCETIN IN SPRAGUE DAWLEY RATS
Vivek Kumar Mishra, Raj Kumar, Anil K. Meena, Pallavi Singh, R.L. Singh & R.K. Singh
ANTIBACTERIALAND ANTIOXIDANT ACTIVITIES OFEUGENIA JAMBOLANUM, BUTEA 52
MONOSPERMAAND CASSIAAURICULATA LEAVES EXTRACT
Kiran Abha Singh, Anup Kumar
REHABILITATION OF SICK INDUSTRIAL UNITS 63
Niaz Ahmed Siddiqui
EMERGING MARKETING STRATEGIES IN TEXTILE INDUSTRIES 73
IN INDIA WITH SPECIAL REFERENCE TO BRAND BUILDING
Monika Yagnik Merh
DEVELOPMENT AND SCOPE OF TOURISM'S SECTOR IN UTTAR PRADESH 78
Rashmi Mishra, Kamlesh Kumar Shukla, Ishvinder Singh Ahluwalia
VARIOUS ASPECTS OF MARKETING OF FINANCIAL SERVICES 83
Kumar Kautilya
RE-ORIENTING THE ROLE AND REVAMPING OF FOOD CORPORATION OF INDIA 87
Shantanu Kumar Srivastava

9. GEOMETRICALAND CONFORMATIONAL STERIC AFFINITY
INTERACTION STUDIES OF ANTICANCER DRUGS WITH
HEXANUCLEOTIDE SEQUENCES OF DNA BY FLUORESCENCE
AND ABSORPTION SPECTROPHOTOMETRY.
*Sulabh Kharbanda
Department of Biotechnology, Saroj Institute of Technology and Management, Lucknow, India
Aromatic ring derivative molecules like
daunomycin, epiadriamycin and adriamycin are
anticancerous anthracycline drugs that are obtained
from Streptomyces peucetius strains of bacteria
through the process of fermentation.These drugs
are effective in the treatment of acute
lymphoblastic leukemia, and myelogenous
leukemia. These drugs intercalate in the major
groove and minor groove of various DNA
sequences and inhibit the replication of DNA
during the S-phase of cell cycle. The structure of
these drugs consists of a planar, hydrophobic
tetracycline ring structure linked to a sugar moiety
with a glycosidic bond linkage. The anthracycline
derivative has four fused rings (A-D).
Epiadriamycin molecule has 4' hydroxyl (OH)
group inversion on the sugar moiety as compared to
Adriamycin and Daunomycin molecules.There are
various forces between drug-DNAcomplexes such
as electrostatic forces, hydrogen bonds,
hydrophobic effects, salt bridges, and dispersion
forces. Electrostatic forces depend upon the
concentration of salt solutions as the concentration
increases, the electrostatic force decreases. The
electrostatic forces vary where there is a presence
ofwaterandor itsabsenceandtheforcesaremuch
*Address for correspondence: Dr. Sulabh Kharbanda, Assistant Professor, Saroj Institute of
Technology and Management, Lucknow, India. Email ID: sulabhchandra2000@gmail.com
ABSTRACT
The anticancer drugs like adriamycin,epiadriamycin and daunomycin intercalate with different
sequences of DNA and provide hindrance to the replication of DNA in cancerous cells by interacting with
DNA gyrase, DNA helicase and DNA topoisomerase which help in the winding and unwinding of DNA
respectively. The drugs bind in the major groove and minor groove of DNA according to the base
specificity of nucleotides in DNA. Various forces that help in the stabilization of drug with DNA are
electrostatic forces like salt bridges, dipole forces like hydrogen bonds, entropic forces like hydrophobic
effect, base stacking forces like dispersion forces. Adriamycin has a higher affinity, cooperativity and
conformational stability during binding to poly(dA.dT) than epiadriamycin and daunomycin Further
studies were done to study the binding of daunomycin, adriamycin, and epiadriamycin to hexanucleotide
sequence d-CGATCG by absorption and fluorescence measurements and energy minimization studies
were done with AMBER force field to study the hydrogen binding interactions between drug and DNA
sequence. In addition to these studies,the affinity of binding of hexanucleotide sequences like d-CGATCA,
and dCGTACA to drugs adriamycin, epiadriamycin and daunomycin was studied and compared with d-
CGATCG sequence. Sugar moiety in drug molecules interacts more strongly with d-CGATCAthan with d-
CGTACAdue to the formation of direct hydrogen bonds between d-CGATCAand not in altered nucleotide
conformation sequence d-CGTACA. In sequence d-CGATCG interaction with drug epiadriamycin, the
4'hydroxyl group is inverted in the sugar molecule as compared to the adriamycin drug where the 4-
hydroxyl group position helps in the formation of hydrogen bonds with adenine N atom in the DNA
hexanucleotidesequenced-CGATCG.
Keywords- Hexanucleotide drug; DNA; Absorption spectroscopy; Fluorescence spectroscopy;
Adriamycin;Epiadriamycin;Daunomycin;Anticancerdrugs; Intercalation;Drug-DNAbinding.
International Journal of Scientific and Innovative Research 2016; 4(2) : 1‐8
P‐ISSN 2347‐2189, E‐ ISSN 2347‐4971
INTRODUCTION
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10. stronger in the absence of water molecules due to
the dielectric constant and polarity of water which
forms hydrogen bonds and weakens the
electrostatic forces. The C-11 H–atom of hydroxyl
groups in ring C forms a hydrogen bond with ring B
oxygen atom in drug molecules and the C-9
carbonyl oxygen atoms in ring D of the drug
molecule also forms hydrogen bonds with guanine
NH –group (amine) nucleotide group in the DNA2
molecule. The other effects include the
hydrophobic effects caused due to the hydrophobic
aromatic anthracycline rings of drug molecules
when water molecules surround them. These water
molecules are released during the formation of
clusters and stacking of aromatic drug molecules
leading to the formation of aggregates. Further
these interactions stabilize the entropy of the
system and decrease the free energy. Further
interaction studies were studied in the intercalation
[1]
of salmon sperm DNA with daunomycin .The
effect of increase in the concentration of drug
daunomycin was studied by spectrophotometric
absorption spectroscopy to analyze the affinity of
intercalation of this drug with salmon sperm DNA
sequence and the concentration of bound, unbound
drug and extinction coefficients of the free and
DNA bound drugs like daunomycin was analyzed
[2].
experimentally
Absorption spectrophotometry
The results of the intercalation of the drug
daunomycin with salmon sperm DNA was
[ 2 ]
analyzed experimentally by absorption
spectrophotometry at wavelength 480nm in 10mM
0
MES buffer, pH=6.2 at 25 C. Further linear plots
and scatchard plots were drawn for the
spectrophotometric data according to the equation
given below, n=n -(1/K )(n/D ), wherem a x a f
n=number of moles of drug bound per mole of
DNA sequence, n = represents the maximummax
binding capacity of the drug with DNA, K =a
association constant for the drug-DNA complex,
and D = is the concentration of unbound drug.f
Further hydrogen bonding of the amino group
present at the 3' position of the sugar moiety in the
drug molecule with the nucleotide of the salmon
sperm DNA was studied and the substitution of the
amino group at the 3' position of the sugar of drug
molecule with peptides lead to an alternation in the
binding affinity of the drug with DNA, and
weakened the interaction and intercalation of the
drug with DNA. Further absorption experiments
were done to study the intercalation of daunomycin
[3]
drug with DNAsequence .The total concentration
of the drug was calculated from the Lambert-Beer's
Law at 540nm wavelength, C = A / wheret 540 540
A =absorbance at 540nm, ϵ = molar extinction540 540
[3]
coefficient at 540nm . Further formula was used to
calculate the amount of bound drug= C =b
A/∆ =(A -A )( - ), the amount of freeo observance f d
drug was calculated by taking the difference of total
drug concentration (C ) – the bound drugt
concentration (C ),A is the absorbance of drug at ab o
known concentration and A is the absorbanceobserved
at a particular ratio of DNA to drug. Further for
absorption studies the concentration of free drug
was calculated according to formula C =C [(A/A -f t o
P)/(1-P,) where C is the concentration of free drugf
C is the concentration of bound drug, A is the, t
absorbance at a given ratio of nucleic acid binding
to drug, A is the absorbance of drug ato
concentration 10µM , P=A /A where A is theα o α
absorbance when all the drug is bound to DNA
sequence. In addition to these experiments
fluorescence studies were done at wavelength
λ =480nm and λ =555nm. Ratio ofexitation emmission
Fluorescent intensity of drug daunomycin before
and after binding to DNA sequence was used to
calculate the amount the concentration of bound
drug to DNA, C =C (I/I -P)/(1-P), where C is thef t 0 t
concentration of bound drug and P=is the ratio of
quantum yield of fluorescence of total bound drug
to DNA to the fluorescent intensity of bound drug
at concentration 10 µM, where P=I /I Analysis ofα o..
data was done ahead by drawing the graphs of r/Cf
where r=number of moles of bound daunomycin
per mole of DNA base pair and C is thef
concentration of free drug(unbound drug), The
calculation of this ratio was explained by the
algorithm loop of Crothers (1968) and closed form
of Mc Ghee and Von Hippel (1974), r/C =K(1-f i
n-1
nr)[(1-nr)/{1-(n-1)}] , where K is the intrinsici
binding constant and n is exclusion parameter in
base pairs. The experiments were continued ahead
by analyzing the equilibrium binding of
daunomycin and adriamycin with calf thymus
[4]
DNA . The absorbance spectra of daunomycin
drug in free state and bound state from wavelength
480nm to 505 nm shows a red shift and
fluorescencespectrawas calculatedat555nmand
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11. 592nm for excitation and emission of the drug
daunomycin . The concentration ratio of drug to
DNA varies from 1:6 to 1:10 along with the
variation in temperature and ionic strength of salts.
Various plots were studied and analyzed for the
binding of drug to DNAby calculating the ratios of
r/C with r at various concentrations of drug from 3-f
0 0
7 µm at different temperatures of 15 C to 25 C. The
sigmoid curves were plotted with following
characteristics of r and free drug concentration (C )f
with drug concentration in the range from 4.4 µm to
5.5 µm for adriamycin. The result suggested that
the sigmoid curve decreased as the concentration of
[4]
the adriamycin decreased and next graphical
analysis of fractional drug saturation and DNA
concentration revealed that as the concentration of
daunomycin was increased, the sigmoid curve of
[4]
fractionalsaturationdecreased.
Cooperativity behavior of the binding of
anticancer drugs with different
sequences of DNA
Experiments suggested that the binding of
adriamycin to DNA takes place by a positive
cooperative mechanism and a positive slope is
formed in the curve of saturation kinetics of drug-
[5]
DNA interaction and is dependent upon the ionic
concentration of salts in the solution. Various
sequences of DNAnucleotides were compared and
it was shown that the positive cooperativity
decreases in the order as follows poly(dG-
dT)—poly(dA-dC) > poly(dAdT)---poly(dA-
[5]
dT),>poly(dG-dC)---poly(dG-dC) . The binding
of the drug decreases with the increase in the salt
concentration from 0.1M to 0.2M. Studies revealed
that daunomycin molecules interact immediately
upstream of the DNA. Further the interaction of
drug daunomycin with sequence dCGATCG has
more pronounced stacking pattern than interaction
of sequence dTGATCA with daunomycin drug.
Studies revealed that daunomycin binds to DNA
specific nucleotides for every two base pairs, and
the binding affinity varies for the drug as it was
observed for guanine nucleotide along with 5'-
methyl group cytosine. The X-ray crystal structure
was developed after the intercalation of drug
daunomycin-(CGTACG) with 2-NH amine2 2
group with variation in ionic strength and
polyelectrolyte concentration with different ionic
[ 8 ]
strength . Crystallization studies of 4'-
epiadriamycin with hexamer dCGATCG revealed
that one drug molecule binds at each CpG site and
the sugar molecule attached to the anthracycline
ring of drug binds to the minor groove of DNA
whereas spermine molecules bind to the major
[9]
groove of DNA. There is variation in the binding
affinity of drug with hexamer sequence dCGATCG
than with sequence dCGTACG. Direct hydrogen
bonds are formed in the drug-dCGATCG complex
and not in drug-CGTACG complex in the minor
groove of DNA. In the major groove of DNA, the
sequence dCGATCG forms a complex with
[6]
spermine protein but not in dCGTACG sequence .
The methylene and amino groups of spermine lead
to the formation of Van-der-Waals forces in the
drug-dCGATCG complex but not in drug-
[6]
dCGTACG complex .Studies ahead proved that
adriamycin drug has much more binding affinity
for hexanucleotide d(TATATA) than for hexanuc-2
leotide d(CGCGCG) although in hexanucleotide2
d(CGTACG) strongest affinity for this sequence is2
observed than for other nucleotides along with
affinity of interaction at the 9'OH position in drug
[10]
and the DNA sequence . Base pair triplet in the
hexanucleotide sequence defines the affinity of
interaction of the drug-DNA complex than the
presence of different types of two base pairs during
drug-DNAintercalationstudies.
MATERIALSAND METHODS
Calf Thymus DNA and hexanucleotides
d(CGATCG) were intercalated with adriamycin,
epiadriamycin and daunomycin anticancer drugs
and the complex was mixed and the results were
analyzed by spectrophotometer and spectro
fluorometer. The concentrations of drugs were
taken in the range of 10µM. The concentration
was100µM for spectrophotometry and 50µM for
spectrofluorometry studies (DNA).0.1M
phosphate buffer was used at pH=7.1 with 5mM
EDTA which was diluted to 20mM phosphate
b u ff e r a n d 1 m M E D TA . A b s o r p t i o n
spectrophotometry was performed on Carry 100
Bio(Varian) UV-visible spectrophotometer with
peltier thermostatted cell holder and quartz cuvette
(path length=1cm).Further titration was done by
mixing 20µl of DNA to a drug solution 9-11µM
and recording the spectrum in the wavelength range
of 200nm-800nm after each addition.
Spectrofluorometrywas performedby adding3ml
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12. RESULTS AND DISCUSSION
1. Absorption spectrophotometry of DNA sequence dCGATCG with drug Adriamycin
The interaction of drug adriamycin with calf
thymus DNA was studied by absorption
spectrophotometry. The kinetics of intercalation of
the drug with DNAwas studied and the wavelength
scanning showed a change in the absorption peak
from 480nm to 503 nm, called as the bathochromic
(red shift). Further isosbestic point was observed at
540nm which is the point on the absorption curve at
which the concentration of bound drug and
unbound drug is the same. Scatter plots were
plotted for r and r/C values as already discussed.f
Absorption spectrophotometry graphs were plotted
for intercalation of adriamycin drug with sequence
dCGATCG(Fig no.1, Fig no.2), graphs for
epiadriamycin drug with sequence dCGATCG(Fig
no. 3,4), graphs for daunomycin with dCGATCG
(Fig no. 5,6) for different concentration ranges and
fluorescent studies were done for the intercalation
of drug adriamycin with sequence dCGATCG,
epiadriamycin with sequence dCGATCG, drug
daunomycin with sequence dCGATCG, and all
these fluorometric intensity graphs showed the
same results as the spectrophotometric graphs
alreadydiscussed.
It was analyzed from the graphs that the binding of
the drugs adriamycin, epiadriamycin, and
daunomycin with calf thymus DNA observes
noncooperative kinetics and the binding of the
drugs with sequence dCGATCG observes
cooperative kinetics The binding sites for the
intercalation of epiadriamycin with DNA was
found out to be maximum, then for daunomycin
and least for adriamycin whereas the association
constant i.e the strength of the binding of drug is
maximum for daunomycin ,then for epiadriamycin
and least for adriamycin as compared to the other
research works with different concentration limits
of drug and sequences of nucleotides of
[10]
DNA .The work can be compared to previous
works where the affinity and the strength of binding
of the anticancer drug adriamycin was analyzed
[10]
with different sequences of DNA . In the graph of
intercalation of drug adriamycin with DNA
sequence dCGATCG, the number of binding sites
of drug to DNA is equal to 0.52 and the Y-axis
depiction of n/C value which is equal to n/ Kf association
6
is2.14x 10 .
www.ijsir.co.in4
Fig- no.1 Graph plot of 1/DNA (dCGATCG) Vs 1/A -A of drug Adriamycin.0
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13. Fig no.2 Scatter plot of r (x-axis) Vs r/C (y-axis)for dCGATCG with drug Adriamycin.f
2. Absorption spectrophotometry of DNA sequence dCGATCG with drug epiadriamycin .
Fig no. 3 Graph plot of 1/DNA(x-axis) (dCGATCG) Vs 1/Ao-A(y-axis) with epiadriamycindrug.
In the same way graph of intercalation of
epiadriamycin with DNA sequence was studied by
absorption spectrophotometry (Fig no. 3, 4).The
number of binding sites of drug on DNA is 0.63,
and the Y-intercept (Fig no. 4) gives the value of
6
r/C whichis equalton/K equalto2.73x10 .f association
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14. www.ijsir.co.in6
Fig no.5 Graph plot of 1/DNA (dCGATCG) Vs 1/A -A of drug daunomycin.0
Fig no. 4 Scatter plot of r Vs r/C for dCGATCG with drug epiadriamycin drug.f
Further absorption spectrophotometry was performed by mixing daunomycin drug with DNA sequence
dCGATCG was calculated to be 0.52. The Y-axis value from fig no. 6 which gives the value of r/C (orf
6
n/K ) is equal to 4.72 x 10 .association
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15. 3. Absorption spectrophotometry of DNA sequence dCGATCG with daunomycin
Fig no. 6 Scatter plot of r Vs r/C for dCGATCG with drug daunomycin drug.f
Further spectrofluorometric quenching studies
were done for the anticancer drugs adriamycin,
epiadriamycin and daunomycin as depicted in the
graphicalfigure no. 7,8.
Fig no 7. The graph shows the ratio of DNA/Drug concentration to the percentage (%)
fluorescence intensity.
[7]
Studies were compared with intercalation of daunomycin with poly(dG-dC) sequence .
Fig no.8 Graph of 1/DNA (x-axis) concentration of dCGATCG and % fluorescence quenching
of drugs.
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16. CONCLUSION
The paper discusses about the strength, affinity,
steric and conformational interaction of anticancer
drugs like adriamycin, epiadriamycin and
daunomycin with different hexanucleotide
sequences of DNA by absorption spectroph-
otometry and fluorescent spectroscopy techniques.
The drugs bind to the major groove and minor
groove of the DNA as it is elucidated in this
scientific article and prevent the replication of
DNA by intercalating with it and further inhibiting
the activity of enzymes like gyrase, topoisomerase
inthecells.
ACKNOWLEDGEMENTS
Author is highly grateful to the faculty members,
Head of department and research scholars of Indian
Institute of Technology, Roorkee for providing the
necessaryinputsforexperiments.
REFERENCES
1. Gabbay E.J., Grier D, Fingerle R.E., Reimer R.,
levy R, Pearce S.W., Wilson W.D., (1976)
Interaction specificity of the anthracyclines
with deoxyribonucleic acid, Biochemistry,
May18, 15(10),2062-2070.
2. Hyman W. R and Davidson N., (1967), The
binding of actinomycin to crab dAT ; the nature
of the DNA binding site, Biochemical and
Biophysical Research communications, 26(2),
116-120.
3. Chaires B.J., Dattagupta N., Crothers D.M.,
(1985) Self-association of daunomycinAugust,
21(17), Biochemistry,3927-3932.
4. Barcelo F., Martorell Jordi., Gavilanes F.,
B,Gonzalez-Ros J.,(1988) Equilibrium binding
studies of Daunomycin and Adriamycin in calf
thymus DNA, Biochemical Pharmacology,
37,11,2133-2138.
5. Graves D.E., Krugh T.R., (1983) Adriamycin
and daunorubicin bind in a cooperative manner
to deoxyribonucleic acid, Biochemistry 2,
22(16),3941-3947.
6. Frederick C.A.,Williams L.D., Ughetto G.,Van
der Marel G.A., van Boom J.H., RichA., Wang
A., (1990) Structural comparison of anticancer
drug-DNA complexes: adriamycin and
daunomycin,Biochemistry,29, 2538-2549.
7. Chaires J.B.,(1986), Allosteric conversion of Z
DNA to an intercalated right-handed
conformation by daunomycin, Journal of
Biological Chemistry, July 5, 261(19), 8899-
8907.
8. Xodo L.E., Manzini.G., Ruggiero.J., and
Quadrifoglio.F.,(1988), On the interaction of
daunomycin with synthetic alternating DNAs:
Sequence specificity and polyelectrolyte on the
intercalation equilibrium, Biopolymers,
November,27(11),1839-1857.
9. Rich A., Ughetto G., Frederick C.A., Williams
L.D.,(1990), Ternary interactions of spermine
with DNA:4' epiadriamycin and other DNA:
anthracycline complexes, Nucleic Acid
Research,18(18),5533-5541.
10. Pullman B., Gresh N., Chen K.S., (1986) A
theoretical investigation on the sequence
specific binding of adriamycin to double
stranded polynucleotides, Nucleic Acid
Research,February,14(5),2251-2267.
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17. www.ijsir.co.in 9
PREVALENCE AND STUDY OF DIFFERENT STAGES OF
CHRONIC KIDNEY DISEASES (CKD) IN PATIENTS.
1 1 1 1
Shailza Verma , Dheeraj Upadhyay ,Rahul Singh ,Neha Pandey ,
1 1 2
Saurabh Singh ,Vishwajeet Pandey ,*Sulabh Kharbanda
1
Graduate Student,Department of Biotechnology, Saroj Institute of Technology and Management,
2
Lucknow, Uttar Pradesh, India, Assistant Professor, Department of Biotechnology, Saroj Institute of
Technology and Management, Lucknow,Uttar Pradesh, India.
Chronic kidney disease (CKD) is prevalent in
many countries of the world like USA, India
etc.The major causes of chronic kidney diseases
[22]
include lack of iron in diet which causes anemia ,
[1]
hypertension, diabetes and stress .Patients were
divided into groups in which one group was given
oral dose of iron(ferritin) and second group was
given intravenous dose of iron. The levels of
albumin, hemoglobin, proteinuria, transferrin,and
glomerular filtration rate(GFR) were unchanged in
both the groups of patient samples analysed by
plasma iothalamatemeglumine method but the
levels of serum ferritin and logarithmic-ratio of
concentration of protein in urine andcreatinine
were changed and increased and decreased for oral
group and intravenous administered iron group
respectively. Various infected samples from skin,
bone, lung, sepsis, urinary tract infections (UTI),
cardiovascular diseases, angina, stroke, and cancer
related patient samples.The rate values for the ratio
of intravenous iron to oral dose of iron and p-values
were reported to be as follows for each sample
(depending on the number of samples in each group
which were adjusted accordingly like skin-3.79,
and p value-0.013, lung value-4.53,and p-value-
0.022, bone value-0.59, and p-value 0.2 (values
adjusted further), urinary tract infections value(less
number of samples)-2.37, and p-value-0.2
(samples-less and less harmful effects), sepsis
(sample-more and more harmful effects) value-
2.59, p value-0.056, cardiovascular-value-2.51,p-
value-is equal to-0.001,arrhythmias value-1.29, p-
value-0.2,angina-value-1.03,p(statistical)-value-
0.2, hyperkalemia- value-0.69, p-value-0.2, cancer
[1]
related samples value-2.07, p-value-0.2 . A
comparative study was done for the analysis of
CKD in patient samples from 48 hospitals and
consisting of 4712 testing specimens. The samples
were analysed for different types of diseases
associated with kidney damage as mentioned in
[2]
Table1 below .
*Address for correspondence: Dr. Sulabh Kharbanda,Assistant Professor, Department of
Biotechnology, Saroj Institute of Technology and Management, Lucknow, India.
Email ID: sulabhchandra2000@gmail.com
ABSTRACT
Chronic kidney diseases are quite prevalent in USA and western countries due to hypertension and
diabetes. Majority of the population in USA and western countries is affected by chronic kidney diseases
leading to increased levels of creatinine and albumin in blood due to increase in filtration pressure inside
the juxtaglomerular apparatus of kidney. In India also the rate of disease affliction has increased
tremendously to 7%, with a range varying from 5%-13% in various regions like Jharkhand and
Chandigarh.The symptoms of kidney disease include changes in the color of urine to darker in
appearance with froth due to the presence of albumins as a result of increase in porosity of basement cell
membrane cells and podocytes with changes in pressure affecting the levels of antidiuretic hormone,
angiotensinogen and renin in kidney along with cirrhosis and the appearance of blood in urine. In
addition to this there is retention of fluid in legs, feet, face, hand and ankles of patients suffering from
chronickidneydiseases.
Keywords:CKD; Nephritis; Nephropathy; Anemia; GFR; Biomarker; Cell Signaling; Transplant.
INTRODUCTION
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18. SNo. Disease name Percentage (%)
1. Diabetes 41%
2 Hypertension 22%
3 Chronic glomerular nephritis 16%
4 Chronic interstitial disease 5.4%
5 Ischaemic nephropathy 5.4%
6 Obstructive uropathy 2.7%
Table 1: The percentage of occurrence of different diseases associated along with kidney
[2]
damage in patient samples in A.I.I.M.S, New Delhi, India .
REVIEWOFLITERATURE
Diagnosis ofChronicKidneyDiseases
The diagnosis of chronic kidney diseases
includesimbalances in the concentrations of ions,
metabolites and hormones in the blood, and the
kidney and changes in the glomerular filtration
pressure (GFR).Adecrease in the concentration of
iron causes anemia and damage to the kidney along
with decrease in the secretion of erythropoietin.
There is increase in the levels of phosphate in
kidney leading to a condition of hyperpho-
sphatemia and less secretion of phosphate in blood,
changes in the concentration of calcium,
parathyroid hormone due to alteration in the levels
of parathyroid hormone leading to bone mineral
metabolism imbalance. Further hypertension is
observed in patients with CKD which further leads
to cardiovascular disease with altered blood
pressure. Diabetes also leads to the development of
[3]
this disease .The high levels of glucose metabolite
in blood leads to changes in the glomerular
filtration pressure in juxtaglomerular apparatus of
kidney, loop of henle, distal convoluted tubule and
proximal convoluted tubule. The alterations in the
levels of ions like sodium, potassium in the blood
and kidney due to the impaired transport in the
aquaporin membrane channel in kidney cells also
leads to the development of kidney diseases due to
the impaired osmolality concentration gradient.
The levels of urea,creatinine and albumin are also
altered in kidney diseases.The renin-angiotensin-
aldosterone system regulates the blood pressure in
the arteries and filtration pressure in the kidney
[ 4 ]
which gets altered in kidney diseases .
Angiotensin converting enzyme (ACE-1) converts
the angiotensin-I to angiotensin-II which constricts
the blood vessels in the kidney leading to changes
in the filtration rate of different metabolites like
glucose, maltose, sucrose, albumin, urea, uric acid,
hippuric acid and creatinine. ACE-1 inhibitor
drugs and angiotensin receptor blockers have been
used to treat hypertension due to altered blood
[4]
pressure .The flow chart depicted in figure no.1
gives the detailed analysis of the changes in
filtration pressure in kidney cells(nephrons).The
secretion of angiotensin and aldosterone in the
renal cells leads to the constriction of blood cells
which alters the filtration pressure in kidney
juxtaglomerular apparatus cells. This mechanism
leads to an alteration in the balance of sodium,
potassium ions across the membrane channels of
nephrons and increase in the absorption of sodium
ions and excretion of potassium ionsbetween
[5]
blood, interstitial fluid and tubules of kidney cells .
There is absorption, assimilation and excretion of
different ions and metabolites in different parts of
kidney namely, loop of henle, proximal convoluted
tubule, distal convoluted tubule and the collecting
duct before they are excreted in the form of urine
fromtheurinarybladder.
Vasoconstriction of blood vessels in kidney due to renin-angiotensin and lack of properfunctioning
of endothelialcellsliningblood vessels due to generationof freeradicals.
Secretion of angiotensin by kidney cells and endothelin-1 by blood vessels.
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19. Changes in glomerular filtration pressure in juxtaglomerular apparatus in kidney due to diabetes, hypertension,
anemia in patients, metabolic-acidosis, bone and mineral changes in calcium and phosphate leading to metabolism
disorder,and accumulationofuricacid,albumin inurine.
Damage to kidney cells (nephrons) leading to cell death and changes in sodium,potassium transport of ions from
membranesof kidney cellstointerstitialfluid and blood.
Figure 1:The flow-chart depicting the changes in the filtration pressure in kidney cells (nephrons).
Differentstages inthedevelopmentof chronickidney diseases.
The different stages of progression of kidney disease are mentioned in table no.2 below along with decrease
[3]
intheglomerularfiltrationrate .
SNo.
1.
2.
3.
4.
5.
Stage of kidney
disease
Stage 1
Stage 2
Stage 3
Stage 4
Stage 5
Glomerular filtration rate volume
2
90ml/min per 1.73m and albuminuria
2
60ml/min-80ml/min per 1.73m
2
30ml/min-59ml/min per 1.73m
2
15ml/min-29ml/min per 1.73m
2
Less than 15ml/min per 1.73m with end stage renal disease
Table 2: The stages of progression of kidney disease along with glomerular filtration rate. Major
types of Kidney diseases leading to chronic destruction of nephrons.
Granulointerstitialnephritis(GIN)
In this condition, there is inflammation at the site of
infection inside the nephrons which leads to the
aggregation of giant cells with multinucleated
condition. There is formation of granuloma inside
the nephrons with accumulation of eosinophils at
that site. Prednisolone drug is used for the
[6]
treatmentof thisnephritiscondition .
Acuteinterstitialnephritis(AIN)
In this disease condition, the interstitial cells of
kidney develop edema, which is the accumulation
of fluid along with inflammation of these cells. The
skin develops rashes, fever and destruction of
nephrons by bacteria. Mycophenolate mofetil drug
[7]
is used for thetreatmentof thisdisease .
Chronictubulointerstitialnephritis.
In this condition, there is destruction of glomerulus
cells of the kidney leading to apoptosis of these
cells. They are caused by bacteria,fungi,virus or
protozoan infection in kidney cells.It is identified
by observing inflammation, interstitial fibrosis, and
apoptosis, atrophy and degradation of proximal and
distal tubular cells. Skin rashes, fever develops in
patients with increase in eosinophils count in
[8,21]
blood.Itis alsocausedby heavymetaltoxicity .
IgAnephropathy
In this condition, there is deposition of IgA
antibody and C3 protein in the mesangial cells of
the kidney and along the walls of glomerulus cells.
In this there is a defect in the O-glycosylation of
IgA antibody during post-translational modifi-
cation of IgA chain and IgA-IgG aggregates are
formed by the recognition of these epitopic regions
on IgA by the immune system of the body leading
to the generation of IgG antibodies complexed with
IgA in the kidney cells leading to destruction and
apoptosis of kidney cells. Galactosyltransfease
enzyme is responsible for the modification and
addition of O-linked carbohydrate groups to the
IgA antibody. The complement system C3-C4-C5-
C9 gets activated along with the release of tumor
necrosis factor (TNF-alpha),TGF-beta, monocyte
chemokine factor-1, macrophage migration
inhibitory factor, interleukin(IL-6) which
promotes inflammation at the site of injury. There
are clusters of CD71-IgA complex, CD89-IgA
complexes formed in the mesangial cells in the
[9]
kidney . Drugs used for treatment of this disease
[10,20]
includecyclosporins .
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20. Lupus nephritis
It is caused by an auto-immune disease called as
systemic lupus erythematosus which causes
damage to the kidney cells. The symptoms include
blood in the urine, frothy appearance and bubbling
in urine and pain and swelling in the hands and legs
of thebody.
Focalsegmentalglomerulonephritis.
In this disease, lesions are formed which leads to
the destruction of podocytes in kidney cells.
Further crescent shaped pauci-immune glomerular
immune complexes are formed after infection in
kidney cells. The levesl of urokinase plasminogen
activator protein(uPAR) increases after inflam-
mation in podocytes. Rituximab, a monoclonal
antibody binds to the CD20 receptor of B-cells and
is used for the treatment of focal segmental
[17]
glomerulonephritis . Further HIV virus infection
alsocausesglomerulonephritis.
DiabeticNephropathy
In this condition, there is increase in the amount of
urinary albumin concentration from a stage of less
albumin concentration called as microalbuminuria
(20µg/min) to a stage of increase in albumin
concentration and the condition is called as
macroalbuminuria- (200µg/min).There is also an
increase in the amount of blood glucose due to
diabetes and the condition is called as hypergl-
ycemia. Drugs used for the treatment of diabetic
nephropathy include repaglinide and nateglinide.
There is a chance of developing anemia and levels
of low density lipoprotein go down (less than
70mg/dl). Sulodexide and pimagedine drugs
reduce the excretion of proteins and albumin during
[18]
diabeticnephropathy .
Drug-induced nephrotoxicity and chronic
kidney diseases
There are various side-effects and harmful effects
of drugs or medicinestaken during infectioncaused
to the body. The side-effects or harmful effects
caused by drugs in kidney are listed in table no.3
[11]
givenbelow .
SNo. Type of Drug Disease caused in kidney
1 Aminoglycosides Chronic tubule- interstitial nephritis
2 vancomycin Tubular destruction
3 acyclovir Crystal formation
4 AmphotericinB Renal tubular acidosis, necrosis
5 ciprofloxacin Needle crystals
6 Pencillins and cephalosporins Toxic tubular injury
7 Antiretroviral drugs Crystalline deposition
8 cetuximab Renal magnesium wasting and hypomagnesia.
9 pamidronate Focal segmental glomerulosclerosis
Table 3: Drug induced nephrotoxicity and chronic kidney disease
Cellsignaling inchronickidney diseases (CKD)
There are various cell signaling pathways which
are activated during chronic kidney disease
diagnosis in nephrons. The cell signaling molecule
cluster is mentioned in table no.4 as mentioned
[12]
below .It depicts the name of cell signaling
molecule and the receptor signaling pathway
activatedduringtheinfectioninkidney.
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21. SNo. Name of cell signaling molecule Signaling pathway
1 clusterin Tumour growth factor-beta
2 Epidermal growth factor pathway Hypoxia inducing factor(Hif - 1)
3 Lipocalin Hif-1, Lcn2, EGF
4 Hepatocyte growth factor pathway Nuclear factor -kappa B, tumour necrosis factor
alpha (less CKD progression), and apoptotic
proteins like Bcl-2(more CKD progression)
Table 4: Cell signaling molecules in chronic kidney diseases
Biomarkersof chronickidney disease(CKD)
There are various markers used for the analysis of chronic kidney diseases as mentioned in table no.5
[13,14,19]
below .
S.No. Name of biomarker moleculein
kidney disease
1 Podocin
2 nephrin
3 podocalyxin
4 Cystatin
5 lipocalin
6 Kidney injury molecule-1
7 N-acetyl glucosaminidase
8 Beta-microglobulin
9 Albumin, transferrin,
Liver FABP(liver type fatty acid
binding protein)
10 Tenascin,TIMP-1
11 Asymmetric dimethyl arginine
Table 5: Biomarkers involved in chronic kidney disease
Treatmentofkidney diseases
The treatment for end-stage kidney disease
involves continuous renal replacement therapy
(CRRT), peritoneal dialysis, and intermittent
hemodialysis. Peritoneal dialysis method involves
the use of catheter to remove solutes in circulation
in blood from kidney. Intermittent hemodialysis
involves the movement of solutes and ions across
the membrane by means of concentration gradient
in blood and kidney cells. This method involves
hemodialysis, hemofiltration, and combination of
two methods. Various salts like saline, anti-
coagulants like heparin, buffer like citrate, and anti-
coagulant drugs like danaparoid, argatobran and
[15]
nafomostatmesilateisused for renaldialysis .
Various immunosuppressive drugs like tacrolimus
is used during kidney transplantation to prevent
organ rejection and auto-immune response in
[16]
patients .
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22. CONCLUSIONS
[1,20]
1. CKD leads to the development of anemia in
patients which was further studied by
administering iron-sucrose in patients and
measuring the glomerular filtration
[1,20].
rates(GFR)
2. A population survey was conducted in patients
from A.I.I.M.S, New delhi which indicated the
percentage of factors responsible for
prevalence of chronic kidney diseases in the
statistical study and predominant factor for
[2]
CKD was found outtobediabetes .
3. Various others diseases associated with CKD
include parathyroidism, bone-mineral
associated disorder, decreased erythropoetin
synthesis, anemia due to iron deficiency,
calcification of skull, and cardiovascular
[3]
diseasesafterstatisticalstudy inpatients .
4. Various biomarkers indicate the diagnosis of
[13,14,19]
chronickidneydisease inpatientsamples.
5. The chronic kidney diseases are characterized
[6,7,8,9,17,18]
asof varioustypesas discussed .
6. Many different types of immunosuppressive
drugs are used for kidney dialysis, replacement
[10,11,15,16]
andtransplantation .
REFERENCES
1. Rajiv Agarwal, MD, John W. Kusek, PhD, and
Maria K. Pappas, BA,A randomized trial of
intravenous and oral iron in chronic kidney
disease, Kidney Int. 2015 October ; 88(4):
905–914. doi:10.1038/ki.2015.163
2. Dash and Sanjay Aggarwal, Incidence of
chronic kidney disease in India, Advance
Accesspublication11,October-2005.
3. Chronic Kidney Disease and Its Complications
Robert Thomas, Abbas Kanso, and John R.
Sedor, –Prim Care. 2008 Jun; 35(2): 329vii.,
doi: 10.1016/j.pop.2008.01.008.
4. S a m i h H N a s r, J a i r a d h a k r i s h n a n ,
VivetteDD'Agati,Bacterial infection related
glomerulonephritis in adults, Kidney
International Journal ,may 2013, Volume 83,
Issue 5,Pages 792-803.
5. Atlas SA, The renin-angiotensin aldosterone
system: pathophysiological role and
pharmacologic inhibition-,J Manag Care
Pharm.2007 Oct;13(8SupplB):9-20.
6. Nicola Joss, Scott Morris, Barbara Young, and
Colin Geddes Granulomatous Interstitial
Nephritis, Clin J Am SocNephrol2:222–230,
2007,December6,2006.
7. Insara Jaffer Sathick, LadanZand, Afrin N.
Kamal, Suzanne M. Norby, Vesna D. Garovic,
Acute interstitial nephritis: etiology,
pathogenesis, diagnosis, treatment and
prognosis, Vol. 5 Issue 1 |, Jan-Dec 2013 | pp. 1
–20,Nephrologyresearchandreviews.
8. Chronic and tubulointerstitial nephritis, Sergey
V.Brodsky, Tibor Nadasky,Chapter
25,Hepistall'sPathologyoftheKidney.
9. Samuels JA, StrippoliGF,Craig JC, Schena FP,
Molony DA IgA NephropathyJonathan Barratt
a n d J o h n F e e h a l l y, I m m u n i t y a n d
Immunosuppressive agents for treating IgA
nephropathy., Cochrane Database Syst Rev.
2003;(4):CD003965.
10.ChabovaV, TesarV, Zabka J, Rychlik I, Merta
M, Jirsa M Jr,Stejskalova A, Long term
treatment of IgA nephropathy by cyclosporine
A,Renalfailure,Jan22(1):55-62.
11. George Sunny Pazhayattil, AnushreeC Shirali,
Drug-induced impairment of renal function,
International Journal of Nephrology and
RenovascularDisease2014:7.
12.Zeba Khan,Manoj Pandey Role of kidney
biomarkers of chronic kidney disease: An
update, Saudi Journal of Biological Sciences
(2014) 21,294–299.
13.Robert G. Fassett, Sree K. Venuthurupalli,
Glenda C. Gobe, Jeff S. Coombes, Matthew A.
Cooper and Wendy E. Hoy ,Biomarkers in
chronic kidney disease: a review, Kidney
International (2011) 80, 806–821;published
online22 June2011.
14.Sun Young Kim and AreeMoon, Drug-Induced
Nephrotoxicity and Its Biomarkers-
,BiomolTher20(3),268-272 (2012).
15.NeeshPannu, RT Noel Gibney, Renal
replacement therapy in the intensive care
unit,Therapeutics and Clinical risk
management,2005:1(2),141-150.
16. Robles-Piedras AL, Gonzalez-Lopez EH,
Tacrolimus levels in adult patients with Renal
transplant, Proc West PharmacolSoc, 2009,
52:33-34.
17. Agnes Fogo: Causes and pathogenesis of focal
segmental glomerulonephritis,Nat Rev
Nephrol.2015 February;11(2):76–87.
18.Jorge L Gross, MirelaAzevedo, Sandra P
Silveiro,LuisHenriqueCanani,Maria
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23. LuziaCaramori, and Themis Zeimanovitz
Diabetic nephropathy: Diagnosis, Prevention,
and treatment, Diabetes Care, 2005, Jan, 28(1):
164-176.
19. Rajiv Agarwal, Kevin L. Duffin, Dennis A.
Laska, James R. Voelker, Matthew D. Breyer,
and Peter G. Mitchell A prospective study of
multiple protein biomarkers to predict
progression in diabetic chronic kidney disease,
Nephrol Dial Transplant (2014) 0: 1–10,doi:
10.1093/ndt/gfu255.
20. Vecchio M, Bonerba B, Palmer SC, Craig JC,
Ruospo M, Samuels JA, Molony DA, Schena
F, Strippoli GFM, Kidney and transplant group,
Cochranegroup,3,Aug 2015.
21. Chronic Tubulointerstitial Nephritis, Edgar V.
Lerma, Chapter 37 Diagnosis and Treatment,
Nephrology and Hypertension,Access Medical
Online.
22.Sheth Nidhi V, Shaila N,Clinico-
Hemotological studies of Chronic Kidney
disease, International Journal of Science and
Research,2014.
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24. HONEY- GOLDEN LIQUID FOR HEALTH
*Vibha Singh
Department of Oral and Maxillofacial Surgery, K.G. Medical University, Lucknow, U.P. India.
Honey therapy is known as Apitherapy.As per
Rigveda this herb born of honey dripped in honey,
sweetened by honey is the remedy for all injuries.
Let's every wing that blow drops honey , let the
rivers and streams recreate honey let all our
medicine turn in to honey , let dawn and the evening
be full of honey our nourishes this sky above be
full of honey let our trees be honey let the sun be
[1]
honeyletourcows secretehoney.
Honey is one of the oldest traditional medicines
considers to be important in the treatment of several
human ailments. Treatment of infection has been
practiced since the origin of mankind. In most of
the ancient cultures honey has been used for both
nutritional and medical purpose. Honey was used
to treat infected wounds as long ago as 2000 years
before Bactria was discovered to be the cause of
infection. In 50AD Dioscorides described honey as
[2]
beinggood for allrottenandhollowulcers.
In Hinduism the Madhu (Honey) is one of the five
ingredients of Panchamrit the five nectars the other
four are ghee , sugar and butter milk . In the temples
honey is poured over the deities in ritual called
Madhuabhiseka. Jatakarma is performed to
welcome the child in the Hindu family by putting
some drops of honey in the childs mouth and
[1]
whisperingnameofgodintheearof child.
Honey is sweet sticky material produced by bees
following the collection of nectar and honey dew.
Its healing properties have been known for
hundreds of years. It was often used in early
decades of the 20th century but after the Second
World War it was gradually supplanted by more
modern and sophisticated products .Although it
was widely used in treating wounds by ancient
civilizations and it is still utilized in remote
communities for the same purpose. The alternative
medicine branch known as Apitherapy offers
treatment based on honey and other bees product
againstdiseaseincludingbacterialinfections.
In the past 25 years or so more and more studies
have been carried out all over the world to gain a
better insight into the efficacious ingredients of
honey, in the light of current state of knowledge
honey deserves more than our passing curiosity as
clinician we should honey full attention especially
in the hospital environment where the bacteria's
becomeresistanttoantibiotics.
The honey have both anti-inflammatory and
antibiotic properties, antioxidant and have greater
influences on healing wound. Honey has been
proven to create favourable condition in the wound
bed autolytic debridement and presence of
substance that promote and accelerate the healing
[3]
process.
Honey has been a well-known medicament since
ancient times but recently there has been a
resurgenceofinterestinusing honeyinthe
*Address for correspondence : Dr Vibha Singh, Professor, Department of Oral and
Maxillofacial Surgery, K.G. Medical University, Lucknow, U.P. India.
Email ID : vibhasinghraghuvanshi@gmail.com
ABSTRACT
Honey is one of the oldest traditional medicines considers to be important in the treatment of several
human ailments. Treatment of infection has been practiced since the origin of mankind. In most of the
ancient cultures honey has been used for both nutritional and medical purpose. Honey was used to treat
infected wounds as long ago as 2000 years before Bactria was discovered to be the cause of infection. In
50AD Dioscorides described honey as being good for all rotten and hollow ulcers. Honey was often used
for wound dressing in the early decade of 20th century but after second world war it as replaced by more
modern and sophistatcated products despite plethora of literature discribing the healing properties of
honey.
Keywords :Antioxidant;Apitherapy;Wound Healing
INTRODUCTION
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25. management of wounds. The excellent antimi-
crobial properties of honey have been reported in
[4]
variousstudies.
ANTIOXIDANTACTIVITY
The antioxidant activity of natural honey is due to
the presence of a variety of compounds such as
flavonoids namely, apigenin, kaempferol,
pinocembrin quercetin, chrysin, galangin,
hesperetin etc. , phenolic acids such as ellagic, p-
coumaric, caffeic, and ferulic acids), ascorbic acid,
catalase, tocopherols, superoxide dismutase,
[5]
reduced glutathione, amino acids and selenium
The anti-oxidant activity has been shown to
strongly correlate with the phenolic content of the
partucilar honey. Gheldofet. Al. reported a linear
correlation between phenolic content and oxygen
[6]
radicalabsorbancecapacity(ORAC).
In addition to neutralization of free radicals, honey
also exerts its anti-oxidant action by inhibiting the
formation of free radicals, catalysed by metal ions
such as iron and copper. Flavonoids and other
polyphenols, common constituents of honey have
the potential to trap these metal ions in complexes,
preventing the generation of free radicals .Honey
contains both hydrophilic and lipophilic anti-
oxidants which may act at different cellular/
physiologic levels making it the ideal source of
[7]
anti-oxidants.
Given its complex composition and diverse
medicinal properties coupled with its impressive
safety profile, it is not surprising that honey has
found a very significant role in modern wound care.
Several randomized controlled trials, investigating
the effect of honey on minor burns demonstrated
accelerated healing time compared with
conventional dressings, such as silver sulfadiazine
It was even found to be superior to non-
conventional dressings such as amniotic membrane
[8]
and potato peel. Furthermore, Analysis of current
evince also indicates the superiority of honey in
superficial and partial thickness burns therapy
Further a recent Cochrane review reported high
quality evidence that honey dressings heal partial
thickness burns more quickly than conventional
dressings Honey compared with 1% silver
sulfadiazine cream in the treatment of superficial
and partial thickness burns Till date, majority of the
studies conducted speak in favour of using honey
for management of radiation-induced oral
mucositis in head and neck cancer patients
receiving radiotherapy, especially in light of the
fact that the conventional therapies have failed to
offer a comprehensive and effective management
.[9,10,11,12]
for oralcomplications
In a meta-analysis, Cho et. Al. reported oral
administration of honey after radiotherapy could
prevent moderate to severe mucositis and
associatedweightloss.
In their meta-analysis, Song et. Al reported an
overall, pooled relative risk of developing severe
Mucositis to be almost 80% lower in patients
treated with honey compared to patients in the
control groups based on the studies included in the
[13]
review.
ANTIMICROBIAL AND WOUND HEALING
PROPERTIES
Honey selected for medicinal use should be
produced under hygienic condition from traceable
source with minimal contamination by pesticide
antibioticsorpollutants.
The antimicrobial nature of all honey is clearly
demonstrated by the ability to remain unspoiled by
micro organism. The inhibitory effect of pure
honey on various Gram-positive and Gram-
negative bacteria and found that most pathogenic
bacteria failed to grow in honey at a concentration
of 40% and above, In particular, Salmonella
Shigella, Enteropathogenic Escherichia coli and
[4]
Vibriocholera.
Honey can be divided in to those whose activity
was confined to their high sugar content low
moisture content and acidity or those that
exclusively generated low levels of hydrogen
peroxide on dilution or those that retained activity
that was independent of synthesis of hydrogen
peroxideondilution.
The types of honey reported in the literature
peroxidase and non-peroxidase honey the ability to
generate hydrogen peroxide has been shown due to
the oxidation of glucose by glucose oxidase which
is an enzyme secreted by the bees at its deposits
nectar and honey dew in to the hive. Peroxidase
honey is not uncommon. Antibacterial activity of
42 Canadian honey against two bacterial species
shows that all the activity was associated with the
production of hydrogen peroxide. Non peroxide
honey however are less common Manuka honey
from New Zealand and Jelly bush honey from
Australia are two common examples of non-
peroxide honey which are postulated to possess
unidentified active components in addition to the
[14]
productionof hydrogenperoxide.
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26. The wound healing properties of honey are because
of three physical properties a very high
concentration of glucose and fructose (osmotic
action) a low Ph value (3.2 -5.4) and low water
content and four important ingredients honey has
three pharmacological characteristics
antimicrobial, wound healing and anti-
inflammatory. Honey promotes healing by
maintaining an environment which is moist (18%)
water and acidic. Its osmotic properties resulting
from high level of simple sugar (80%) honey helps
clear away the necrotic tissue in the wound bed .
This prevents the dressing sticking to the wound
and protects the granulation edges and this turn
stimulates cell division mitosis, because of this
high glucose fructose concentration honey
deprives the inflammatory bacteria of their
nourishment . Instead of metabolizing the amino
acids the amine and sulphur containing
decomposition products of amino acid causes
putrid smell the bacteria concentrate on the
producing lactic acid and acidifying environment
through this mechanism honey is able to inhibit
development of pathogenic bacteria that causes
infection.
The presence of glucose oxidase enzymes leads to
the formation of hydrogen peroxide in the honey in
accordancewithfollowingchemicalreaction
Glucose + water = gluconic acid + hydrogen
peroxide.
This release very gradually after 12 hours the
concentration is 4 to 5 microgram after 24 hour and
this is sufficient to disinfect wound to set autolytic
debridement in motion and promote granulation
without any risk of toxic effect that would be
causedby toohighconcentrationofhydro radicals.
A second ingredient with antibacterial properties
this is one non peroxide was demonstrated by
Professor Thomas Henle at Germany in 2008. This
is substance which have been known for many
years because it is present in all food stuffs with
higher sugar content Methylglyoxal. This is one of
the dicarbonyl components that are formatted
through Maillard reaction which occurs in all
products that have very high sugar content. It varies
according to the geographic origin and the type of
honey .Depending on the MGO content which can
range from 3-4 microgram to 750-800 microgram
per gram honey the honey will have weaker or
strong effect on a narrower or wider spectrum of
bacteria particularly on the methicillin resistant
Staphylococcus aureus strain, the vancomycin
resistant enterococci and pseudomonas aeruginnisa
whichareunaffectedby presentdayantibiotics.
The third effective ingredient was demonstrated by
Dr Zaat in 2009Aminute quantity of the substance
ranging from 2-3 nanogram per gram honey is
present in all types of honey . This substance is
similar to human beta defensin 1 (HBD-1) protein a
peptide molecule with cationic properties which
play an antimicrobial role by aggregation and
destruction of the host cell behaving like a true
peptideantibiotic.
A fourth group of substance which al so play an
important role in wound healing are the flavonoid a
group of molecules belonging to the polyphenols
which are known to be effective against type 1
radicals.
At high concentrationthese substances reduces any
inflammation present and moderate the pain the
important of these effects during the wound healing
process should not be under rated as they make the
[15]
episodemorebearablefor thepatient.
COMPOSTION
Honey comprise of 40% glucose, 40% fructose,
20% water with organic acid , vitamins enzymes
and minerals. It has specific weight 1.4 and ph 3.4.
Treatment with honey is simple and inexpensive
and it's not needs to be sterilize as it already
possesses a bactericidal property. Because of its
high viscosity it forms a physical barrier creating
moist environment which appears to be helpful and
[16]
acceleratewound healing.
The exact molecular mechanism of wound healing
using honey is yet to be elucidated .various studies
shows that it acts by reducing ROS levels beside
this it exert antibacterial activity and low pH and
high free acid content may assist wound healing
withhoney.
The types of wound and degree of severity al so
affect efficacy selected honey should be used in
sufficient quantity so that it remains there if diluted
with wound exudates .It should cover and extend
beyond the wound margin, the effect of wound
healing is result of combined effect of chemical
debridement of dead and devitalized tissue from
ulcer by catalase, absorption of edema by
hygroscopic properties of honey and promotion of
granulationandepithelisationfromwound edges.
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27. Even raw honey can be used directly for wound
care when all of the above facilities are not
available for sterilization of the honey. Wound care
by honey should be supervised by physician or
experienced wound care nurse in patients with
significant co morbidities. Any co morbidity
problem which contribute to the problem in wound
healing should be diagnosed and treated. It should
be contact with the wound for at least 12 hours,
preferably for 24 hours. If dressing is not
appropriate the honey may be wash out of the
wound byexudates.
In the case of infected wound systemic antibiotic
should be administered after local swab has been
taken or if fever or local sign revealed soft tissue
infection. The use of honey never has been
observedtofosterbacterialrésistance.
There are some adverse effect related to use of
honey. Stinging pain after administration of honey
is reported in 5% of cases, this can be managed by
applying Local anaesthetic cream but it causes
vasoconstriction which can result in reduction in
localperfusion.
It can also be used preoperatively to prepare
wounds for reconstruction or to make it possible to
use smaller or less complex graft. This will help in
reducing post-operative morbidity of the donor
site. The effect of treatment should be evaluated
[17]
continuallyforaperiodofuptotwoweeks.
Types of wound, care protocols and pharmaceutical
requirements for the medicinal use of honey Honey
therapy canal so play a palliative role in improving
quality of life for terminal patients with decubitus
ulcers. It helps in changing dressing without any
pain and al so eliminates unpleasant foul odour .But
if debridement is inadequate honey dressing will
not help because it does not replace good quality
[18]
basicwound care.
There is increase in use of honey as a dressing on
infected wounds burns and ulcers but there is
some common concern that risk of botulism form
clostridial spores sometimes present in honey. It is
established fact that antibacterial activity is heat
labile so would be destroyed if honey will be
sterilized buy autoclaving. So effect of gama
radiation in sterilization of honey and its
antibacterial properties was studied by Molan PC
et al showed that 25 kGy of gama radiation was
sufficient to achieve sterility without affecting its
[19]
antibacterial properties. Use of honey in
management of damaged intestinal mucosa, It
promote the repair of damaged intestinal mucosa
stimulate the growth of new tissue and work as an
anti-inflammatory agent. It also reduces symptoms
of inflammation when applied to the wounds. Raw
honey contains copious amount of compound like
flavonoids and other poly phenols which may
[20]
functionasantioxidants.
The mucositis a side effect of chemo and
radiotherapy that affects the entire gastrointestinal
tract from the mouth to the anus .the cancer
treatment breaks down the epithelial cell lining the
tract leaving the patient prone to ulceration and
infection. The use of honey in oral mucositis
followed by radiotherapy is very effective in terms
[21]
of ulcerhealingandqualityof lifeimprovement.
The use of honey in the cases of alveolar osteitis dry
socket followed by dental extraction and use of
honey in oral mucosistis.. The symptomatic relief
was significant, and results were quite promising
both of the studies were done in the department of
[21]
oralandmaxillofacialsurgeryK.G.M.U.
.Honey has a complex chemical composition and
neither the identities of all of its inhibitory
components nor its mechanisms of action are yet
completely understood. Laboratory tests have
demonstrated the effective inhibition of a wide
range of microbial species, with both antibiotic-
sensitive and antibiotic-resistant bacteria showing
susceptibility.The publication of case reports of the
eradication of MRSAfrom patients give validity to
in vitro observations, but large scale clinical trials
are needed to establish its clinical efficacy. With the
increased availability of licensed wound care
products containing honey, clinical use is expected
to increase and further evidence will become
available. Honey seems to have the potential to
clear infection as well as being an effective
prophylactic agent that may contribute to reducing
the risks of cross-infection. Time will demonstrate
whether the present optimism about honey is
[22]
justified.
The theapeutic effectiveness can be guaranteed by
measuring the peroxidase activity and antibacterial
properties against a selection of bacteria . active
ingradient contents of honey depends on the
production area and the way in which the honey is
collected and processed the bee keeping
production and collection of honey needs to be
carriedoutunderstrictlycontrolledconditions.
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28. It is essential that honey that is produced in
protected natural areas such as Regional Park is
selected and tested for pesticides and heavy metals.
Absence of bactrial contimainartion yeast and
botulism spore. Honey is quickly contaminated
when it is exposed to the air especially in the
polluted atmosphere .Sterlization with gama rays is
neccessary to produce honey with bacterial burden
of no morethan30CFU/gram.
Presence of pollen may cause alllergic reaction in
some patients .In moderate climate regions honey
can be stored at room temperature but temperature
25 degree should be avoided optimum storage
temperature cis between 12-25 degree temperature
. it should al so be protected against UV rays as
there are chances of changed in honey by UV rays.
In more than 500 publications th clinical use of
honey in open wound in the literature no adverse
reaction have been noted . some times a localised
stinging sensation described by some patients .
trhis may be due to acidity of the honey as it has not
[23]
beenreportedwhenacidityis nutrilsed.
CONCLUSIONS
Honey was often used for wound dressing in the
early decade of 20th century but after second world
war it as replaced by more modern and
sophistatcated products despite plethora of
literature discribing the healing properties of
honey. There are many possible reasons why the
pharmaceutical companies has ingnpred this
natural product including lack of knowledge lack
of research in to medicinal use of honey .Honey
seems to be have the potential to clear infection as
well as being in effective prophylactic agent that
may contribute to reducing the risk of cross
infection.
There are some practical consideration which
shoould be tken care of before using honey for
clinical cases. The amount of honey required for
the wound dressing should depends on the amount
of exudate released from wound will result in
dilution of the dressing. If there is no exudate
dressing needs to change twice /week. It should be
applied to absorbent dressing because if applied
directly on the wound it tends to rub off bfore
secondary dressing is applied. Soaking of honey in
to direct dressing facilitiated by warming honey to
body tempertaure or adding one part with 20 part
of honey. For moderately to heavy exudated wound
a secondary dressing may be needed .Honey can be
used to treat cavity wound by using adhesive film
dressing .A low adhrent dressing helps prevent he
honey sticking to the wound but it must be porous
to allow antibacterial component of the honey to
diffusefreelyintothewound bed.
Honey can be safely filled in to cavities and sinuses
. it is water soluble and easily rinse out any any
residue are biodegradable. For sinuses catheter or
syring can be used for applying honey . honey
dressing needs to extend beyond the inflammed
areasurrounding awound.
REFERENCES
1. Nayik et al. UJP 2014, 03 (01): Page 5-8
www.ujponline.com .Universal Journal of
Pharmacy,03(01), Jan-Feb2014 .
2. Gunther RT. The Greek Herbal of Dioscorides
The Greek Herbal of Dioscorides. New York:
Hafner,1934 (reprinted1959).
3. GMS Krankenhhyg Interdiszip. 2007; 2(2):
Doc51.Publishedonline2007 Dec28.
4. Mohammad JavedAnsari,AhmadAl-Ghamdi,
Salma Usmani, Noori S. Al-Waili, Deepak
Sharma, AdgabaNuru, and Yehya Al-Attal ,
Effect of Jujube Honey on Candida albicans
Growth and Biofilm Formation. Archives of
MedicalResearch44(2013) 352e360.
5. Ahmad Oryan, EsmatAlemzadeh , Ali
Moshiri, Biological properties and therapeutic
activities of honey in wound healing: A
narrative review and meta-analysis. Journal of
TissueViability(2016) 25, 98-118.
6. Gheldof N Wang XH , Engeseth NJ
identification and quantification of antioxidant
component of honey from various floral
sources. J.Agric Food Chem 2002 Oct 9 50
(21)5870-7.
7. Subrahmanyam M.Aprospective randomised
clinical and histological study of superficial
burn wound healing with honey and silver
sulfadiazine.Burns 1998;24:157-61.
8. Subrahmanyam M. Honey dressing versus
boiled potato peel in the treatment of burns: a
proDec;51(2-3):121-34.
9. doi: 10.1080/10376178.2016.1171727. Epub
2016Apr Mashhood AA, Khan TA, Sami AN.
spective randomized study. Burns
1996;22:491-3. . J Pak AssocDermatol
2006;16:14-9.
International Journal of Scientific and Innovative Research 2016; 4(2) : 16‐21
P‐ISSN 2347‐2189, E‐ ISSN 2347‐4971
www.ijsir.co.in20

29. 10. Bangroo AK, Katri R, Chauhan S. Honey
dressing in pediatric burns. J Indian
AssocPediatrSurg2005;10:172-5.
11. Subrahmanyam M, Sahapure AG, Nagane NS,
Bhagwat VR, Ganu JV. Effects of topical
application of honey on burn wound healing.
Ann Burns FireDisasters2001;14(3).
12. Subrahmanyam M. Honey-impregnated gauze
versus amniotic membrane in the treatmentof
burns. Burns 1994;20:331-3.
13. Song JJ, Twumasi-Ankrah P, Salcido R.
Systematic review and meta-analysis on the
use of honey to protect from the effects of
radiationinduced oral mucositis. Adv Skin
Wound Care2012;25(1):23-8.
14. GMS Krankenhhyg Interdiszip. 2007; 2(2):
Doc51.Publishedonline2007 Dec28.
15. Dr David LECHAUX, gastrointestinal
surgeon - Hôpital Yves Le Foll 22000 ST-
BRIEUC (France). Previously specialist in
internal medicine at various hospitals in
Rennes (Fr). Chairman of the Food and
NutritionLiaisonCommittee
16. Topical Application of Honey in The
Treatment of Wound Healing: A Metaanalysis
B Medhi, A Puri*, S Upadhyay,** L
Kaman***PMCID: PMC2686636
17. Arne Simon KirstenTraynor Kai Santos Gisela
Blaser UdoBode and Peter Molan Medical
Honey for Wound care Still a latest Resort .A
Evid Based Complement Alternat Med 2009
Jun 6(2) 165-173.
18. Dr David Lechaux, gastrointestinal surgeon -
Hôpital Yves Le Foll 22000 ST-BRIEUC
(France). Previously specialist in internal
medicine at various hospitals in Rennes (Fr).
Chairman of the Food and Nutrition Liaison
Committee.
19. Molan PC Allen K The effect of gama
irradiation on antibacterial activity of honey .
J PharmaPharmacol1996 Nov 48(11)1206-9.
20. Manisha Deb Mandal and Shyam APADA
Mandal -Honey ita medicinal property and
antibacterial activity Asian Pac J Trop
Biomed2011Biomed2011Apr 1(2) 154-160
21. Vibha Singh ,US PAL , Nikita V Soni . Honey
a sweet approach to avlveolar osteitis National
journal of maxillofacial surgery 20144 Jan
5(1) 31 -4.
22. Rose Cooper Centre foe Biomedical Sciences
Cardiff Scool of health Sciences University of
Wales Institute Cardiff . United Kingdom
GMS Krankenhaushyg Interdiszip 2007 2(2)
D o c 5 1 .
http://www.egms.de/de/journals/dgkh/2007-
2/dgkh000084.shtml
23. Honey and wound care dr david lechaux
D.Lechaux Yves Le Foll Hopital St Briuc. D.
Lechaux, Yves Le Foll Hôpital,St--Brieuc
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30. PHENYLHYDRAZINE INDUCED HAEMATOTOXICITY AND IT'S
RECOVERY BY FUMARIA INDICA PLANT EXTRACT IN CHARLES
FOSTER RATS
1 1 1 2 3 1
Raj Kumar , Vivek Kumar Mishra , Anil K.Meena , Pallavi Singh , R.L.Singh and *R .K. Singh
1
Division of Toxicology ,CSIR-Central Drug Research Institute,Lucknow-226031, Uttar Pradesh, India,
2 3
Department of Biotechnology, CEAT, IILM Academy, Greater Noida , Uttar Pradesh, India, Dr.
Ram Manohar Lohia Avadh University, Faizabad, Uttar Pradesh, India
Blood is a body fluid in animals that delivers
necessary substances such as nutrients and oxygen
to the cells and transports metabolic waste products
away from those same cells. In vertebrates, it is
composed of blood cells suspended in blood
plasma. Plasma, which constitutes 55% of blood
fluid, is mostly water and contains dissipated
proteins, glucose, mineral ions, hormones, carbon
dioxide and blood cells themselves.Albumin is the
main protein in plasma, and it functions to regulate
the colloidal osmotic pressure of blood. The blood
cells are mainly red blood cells (also called RBCs
or erythrocytes), white blood cells (also called
WBCs or leukocytes) and platelets. The most
abundant cells in vertebrate blood are red blood
cells. These contain haemoglobin, an iron-
containing protein, which facilitates oxygen
transport by reversibly binding to this respiratory
gas and greatly increasing its solubility in blood. In
contrast, carbon dioxide is mostly transported
extracellular as bicarbonate ion transported in
plasma. The word toxicity is used to describe the
potential for a material to produce injury in
biological system. Phenyl hydrazine is inducing to
formation of reactive oxygen species, and it
changes the haemoglobin in oxihaemoglobin.
Phenyl hydrazine causes lipid peroxidation and
degration of membrane skeleton and cause
haemolyticanaemia.It also decreaseshaemoglobin
level, red blood cell concentration and packed cell
volume, and impairs erythrocyte deformability.
Due to lipid peroxidation, RBCs enter in the
spleen and causes splenomegaly. Methanolic
extract of the plant Fumaria indica contains
monomethy fumarate, which showed significant
protection against hepatotoxicity induced by
carbon tetrachloride,paracetamoland rifampicinin
vivo. The plant extract is used as antibacterial, anti-
implantation anti estrogenic activity and anti-
cancer. The Fumaria indica treats fever and
influenza. Fumaria indica stem is used as a tonic. It
is also used in scrofula, constipation, and jaundice.
It is also used as a component of various herbal
product such as esano capsule and Ayurveda
capsule.
* Address for Correspondence: Dr. R.K. Singh , Chief Scientist & Professor , Division of Toxicology,
CSIR-Central Drug Research Institute, Lucknow-226031, Uttar Pradesh, India,
Email ID : rktox@yahoo.com
ABSTRACT
Phenyl hydrazine (PHZ) is a potent chemical causing haemolytic anaemia in various tissues at various
levels. Phenyl hydrazine is used as a model for the induction of haemolytic anaemia in Charles foster rats.
It causes the destruction of red blood cells by oxidative stress within erythrocytes and changes at cellular
level resulting haemolytic anaemia. Flowers of Fumaria indica showed the remarkable haemo protective
activity against the haemolytic anaemia. Methanolic extract of Fumaria indica showed the
haemoprotective activity against phenyl hydrazine induced haemolytic anaemia in Charles foster rats at
different doses. The plant extract contains this haemoprotective activity due to the presence of high
phenolic,flavonoidandalkaloidcontents.
Keywords: Phenyl Hydrazine (PHZ); Haemolytic Anaemia; Red Blood Cells; Oxidative Stress;
CytochromeP FumariaIndica;MedicinalandPharmacologicalActivities450,
INTRODUCTION
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31. 1. Contents of Blood
The blood of all vertebrates contains the following
contents.
 Erythrocytes(RBC)
 Leukocytes (WBC)
 Platelets (Thrombocyte)
 Erythrocyte(RBC)
Red blood cells also called erythrocytes are the
most common type of blood cell and the vertebrate
organism's principal means of delivering oxygen
(O2) to the body tissues—via blood flow through
the circulatory system. RBCs take up oxygen in the
lungs or gills and release it into tissues while
squeezingthroughthebody's capillaries.
 Leukocytes (WBC)
White blood cells (WBCs), also called leukocytes
or leucocytes, are the cells of the immune system
that are involved in protecting the body against
both infectious disease and foreign invaders. All
white blood cells are produced and derived from
a multi-potent cell in the bone marrow known as
a hematopoietic stem cell. Leukocytes are found
throughout the body, including the blood and
lymphatic system.
 Platelet
Platelet also called thrombocytes are a component
of blood whose function is to stop bleeding by
clumping and clotting blood vessel injuries.
Platelets have no cell nucleus: they are fragments of
cytoplasm that are derived from the megakaryocytes,
of the bone marrow, and then enter the circulation.
These inactivated platelets are biconvex discoid
(lens-shaped) structures, 2–3 µm in greatest
diameter.
2. Phenylhydrazine
Phenyl hydrazine (PHZ) was the first hydrazine
derivative characterized by Hermann Emil Fischer
in 1875. This compound used worldwide mainly as
a chemical intermediate in pharmaceutical,
agrochemical, and chemical industries. PHZ,
C6H8N2 has a molecular weight 108; it exists as
yellow to pale brown crystal or as a yellowish oily
0
liquid, with a freezing point of 19.6 C and a boiling
0
point of 243 C. PHZ metabolism seems to occur
via ring hydroxylation and conjugation, excretion.
PHZ derivatives were used firstly as antipyretics
but the toxin action on red blood cells made their
used dangerous. For many years, phenyl hydrazine
is used for experimental induction of anaemia in
animals until Morawitz and Pratt suggested it as a
drug for polycythaemia Vera (Falconer 1933), a
clonal disorder (Spivak 2002) which is known by a
net increase in a total number of erythrocyte in the
body.
Chemical formula‐ C6H8N2
Molar mass‐ 108.14g/mol
Boiling point ‐ 243.5°C
Mel ng point ‐ 19.5C
Colour‐ Yellow oily Liquid
Fig.1: Structure of Phenyl hydrazeen
1. Mechanism of Phenyl hydrazine (PHZ)
induced haematotoxicity
PHZ reacts with carbonyl group (C=O) which is
common among biological molecule so it directly
attaches with biological molecule. PHZ is taken up
by the inhalation, orally, and dermal roots in
animals and humans. After absorption, PHZ is
rapidly taken up by the red blood cells. In RBCs,
PHZ interacts with hemoglobin and cytochrome
P and causes oxidation reaction and generates450
destructive free radicals that are mainly responsible
for hemolysis.
Phenyl hydrazine
Oxidation of oxy hemoglobin
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32. p–HYDROXYPHENYLHYDRAZINE
(It causes hemolysis and lipid peroxidation of RBCs)
FORMATION OF PHENYL FREE RADICAL (REACTIVE OXYGEN SPECIES)
(Released in to haemoglobin)
CONVERSION OF OXYHAEMOGLOBIN IN TO METHEMOGLOBIN,
HEMICHROMES AND OTHER HEMOGLOBIN BREAKDOWN PRODUCTS
(Oxidative alteration of RBC protein)
HEINZ BODIES FORMATION
(Formation of phenyl radical and the replacement of hemi with phenyl- substituted
protoporphyrin causes destabilization of hemoglobin to induce Heinz Bodies)
UPTAKE OF RBCs BY MACROPHAGES IN SPLEEN
TRANSLOCATION OF PHOSPHATIDYLSERINE FROM THE INNER
TO OUTER OF THE PLASMA MEMBRANE
PHAGOCYTOSIS OF CELL UNDER PROGRAMMED DEATH BY MACROPHAGES
Fig.2: Process of Phenyl hydrazine induced haematotoxicity
1. Haemolytic anaemia
Hemolysis is the type of destructive process in
which destruction or removal of red blood
Cells occurs from the circulation before their
normal life period of 120 days. When the
haemolysis exceeds to be lifelong
asymptomatic condition, it often causes
anaemia. And this particular condition is
known as haemolytic anaemia. Hemolytic
anaemiacanbeextrinsicorintrinsic.
 Extrinsichaemolyticanaemia
Extrinsic hemolytic anaemia is also known as
autoimmune hemolytic anaemia. This type of
anaemia develops when the spleen traps and
destroys healthy red blood cells. It can also
come from red blood cell destruction due to
infection, tumours, autoimmune disorders,
medication side effects, leukemia,
lymphoma.
 Intrinsic hemolyticanaemia
Intrinsic hemolytic anaemia develops when
the red blood cells produced by the body are
defective. This condition is often inherited,
such as in people with sickle cell anaemia or
thalassemia. Anyone of any age can develop
hemolytic anaemia. However, according to the
Na onal Heart, Lung, and Blood Ins tute (NHLBI),
hemolytic anaemia seems to affect more
AfricanAmericansthanCaucasians.
 Causes of Hemolytic Anaemia
Some underlying causes of extrinsic hemolytic
anaemia are enlarged spleen, hepatitis,
Epstein-Barr virus, typhoid, fever,
Escherichia coli, streptococcus, leukaemia,
lymphoma.
 Symptoms of Hemolytic Anaemia
These are some common symptoms
haemolytic anaemia. These are paleness of
the skin, fatigue, fever, light-headedness,
dizziness, weakness or inability to do physical
activity.Therearesomeless commonsigns
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