Indian Journal of Experimental Biology

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VOLUME 47

NUMBER 11

NOVEMBER 2009

CODEN: IJEB (A6) 47 (11) - (2009) 843-932

ISSN: 0019-5189 (Print); 0975-1009 (Online)

 

CONTENTS

 

Review Article

 

Biological activities of crude extracts and chemical constituents of Bael,
Aegle marmelos (L.) Corr.

 

849

      Pallab Maity, Dhananjay Hansda, Uday Bandyopadhyay & Dipak Kumar Mishra

 

 

 

Papers

 

Establishment of CHO cell line expressing human MCHR2 gene and research of its molecular characteristics

 

862

      Chengfu Yuan, Youquan Bu, Joyeeta Sengupta, Junxia Yang, Xiuning Huang,
Li Cheng, Qin Zhang, Faping Yi, Geli Liu & Fangzhou Song

 

 

 

Evaluation of aroA deletion mutant of Salmonella enterica subspecies enterica serovar Abortusequi for its vaccine candidate potential

 

871

      Javed Alam, B R Singh, D Hansda, V P Singh & J C Verma

 

 

 

Neuroprotective effect of MK-801 against intra-striatal quinolinic acid induced behavioral, oxidative stress and cellular alterations in rats

 

880

      Harikesh Kalonia, Puneet Kuamr, Bimla Nehru & Anil Kumar

 

 

 

Chronic prenatal restraint stress induced memory impairment in passive avoidance task in post weaned male and female Wistar rats

 

893

      Saju Binu Cherian, K L Bairy & Muddanna S Rao

 

 

 

Tamarindus indica L. and Moringa oleifera M. extract administration ameliorates fluoride toxicity in rabbits

 

900

      R Ranjan, D Swarup, R C Patra & Vikas Chandra

 

 

 

Solar and artificial ultraviolet-B induced erythrocytes Hemolysis with photosensitizers

906

      Sunil Kumar, Shoma Devi, Prashasti Misra & Priyanka

 

 

 

Quantitative PCR: A quality control assay for estimation of viable virus content in live attenuated goat pox vaccine

 

911

      D J Kallesh, M Hosamani, V Balamurugan, V Bhanuprakash, V Yadav &
R K Singh

 

 

 

Genetic engineering of avian pathogenic E. coli to study the functions of FimH adhesin

916

      H H Musa, S F He, S L Wu, C H Zhu, Z H Liu, Z N Zhang, V S Raj, R X Gu &
G Q Zhu

 

 

 

Notes

 

Protocol for improved extraction and PCR amplification of genomic DNA from liverwort, Plagiochasma appendiculatum

 

921

      Arvind Soni & Anil Kumar

 

 

 

Assessment of genetic fidelity of micropropagated apple rootstock plants, EMLA 111, using RAPD markers

 

925

      R Gupta, M Modgil & S K Chakrabarti

 

 

 

Notes & News

 

Influenza A (H1N1) pandemic: Preparedness and clinical management

929

Madhu Khanna & Neha Gupta

 

 

 

Announcement

 

National Symposium on Recent Advances in Research on Snake Venom and
Snakebite Therapy: National and International Perspectives (SnakSymp-09)

 

848

 

————————

 

Announcement

 

 

National Symposium on Recent Advances in Research on Snake Venom and Snakebite Therapy:
National and International Perspectives (SnakSymp-09)

18 and 19 December 2009, Tezpur

 

Sponsored by the Department of Biotechnology, Govt. of India and the Indian Council of Medical Research, the Symposium will cover following themes: (i) Biochemistry/
bioinformatics/biophysics/molecular biology of snake venom proteins/enzymes/toxins,
(ii) Evolution of snake venom proteins and toxins, (iii) Pathophysiology and treatment of snakebite, (iv) Medical/biotechnological/diagnostic application of snake venom peptides,
(v) Green medicine for snakebite and (vi) Biology of snakes and snakes in India. For details, please contact, Prof. A. K. Mukherjee, Organizing Secretary, or Dr Robin Doley, Joint Secretary, Department of Molecular Biology and Biotechnology, Tezpur University, Tezpur 784 028, India. Telephone: +91 9957 184351 (AKD)/ 91 94357 54830 (RD). E-mail: akm@tezu.ernet.in/
snaksymp@gmail.com; or doley@tezu.ernet.in

 

 

 

Author Index

Alam Javed

871

Anil Kumar

880, 921

 

 

Bairy K L

893

Balamurugan V

911

Bandyopadhyay Uday

849

Bhanuprakash V

911

Bu Youquan

862

 

 

Chakrabarti S K

925

Cheng Li

862

Cherian Saju Binu

893

 

 

Gu R X

916

Gupta Neha

929

Gupta R

925

 

 

Hansda D

871

Hansda Dhananjay

849

He S F

916

Hosamani M

911

Huang Xiuning

862

 

 

Kallesh D J

911

Kalonia Harikesh

880

Khanna Madhu

929

 

 

Liu Geli

862

Liu Z H

916

 

 

Maity Pallab

849

Mishra Dipak Kumar

849

Misra Prashasti

906

Modgil M

925

Musa H H

916

 

 

Nehru Bimla

880

 

 

Patra R C

900

Priyanka

906

Puneet Kuamr

880

 

 

Raj V S

916

Ranjan R

900

Rao Muddanna S

893

 

 

Sengupta Joyeeta

862

Shoma Devi

906

Singh B R

871

Singh R K

911

Singh V P

871

Song Fangzhou

862

Soni Arvind

921

Sunil Kumar

906

Swarup D

900

 

 

Verma J C

871

Vikas Chandra

900

 

 

Wu S L

916

 

 

Yadav V

911

Yang Junxia

862

Yi Faping

862

Yuan Chengfu

862

 

 

Zhang Qin

862

Zhang Z N

916

Zhu C H

916

Zhu G Q

916

 

 

Keyword Index

Aegle marmelos

849

Abortion

871

Amelioration

900

Antibacterial

849

Anticancer

849

Antidiabetic

849

Antifungal

849

Antihyperlipidaemic

849

Antioxidant`

849

Antiulcer

849

Antiviral

849

Apple rootstock

925

Avian pathogenic E. coli

916

 

 

Bael

849

 

 

Chloroquine

906

Cognition

893

CTAB

921

 

 

DNA fingerprinting

925

 

 

Eukaryotic expression vector

862

FimH

916

Fluoride-toxicity

900

 

 

Gene expression

862

Genetic fidelity

925

Genomic DNA

921

Glutathione

880

Goat pox vaccine

911

Guinea pig

871

 

 

H1N1

929

Hippocampus

893

Host cell

916

Huntington’s disease

880

 

 

Influenza A

929

 

 

MCHR2

862

Micropropagation

925

Mitochondrial dysfunction

880

MK-801

880

Molecular characteristics

862

Molecular markers

925

Moringa oleifera

900

Mutant aroA

871

 

 

Oxidative stress

880

Ozone depletion

906

 

 

Passive avoidance test

893

PCR

921

Plagiochasma appendiculatum

921

PPR vaccine

911

Prenatal stress

893

 

 

Quinolinic acid

880

 

 

Rabbit

900

Radioprotective

849

Riboflavin

906

 

 

Salmonella enterica Abortusequi

871

Striatum

880

Swine flu

929

 

 

Tamarindus indica

900

TaqMan QPCR

911

 

 

Ultraviolet-B

906

Vaccine

871

 

 

Indian Journal of Experimental Biology

Vol. 47, November 2009, pp.849-861

 

Review Article

 

 

Biological activities of crude extracts and chemical constituents of Bael,
Aegle marmelos (L.) Corr.#

Pallab Maity1,†, Dhananjay Hansda2, Uday Bandyopadhyay1,†† & Dipak Kumar Mishra3*

1 Division of Drug Target Discovery and Development, 2 Laboratory Animal Division and
3 Botany Division, Central Drug Research Institute, (CDRI), CSIR, Chatter Manzil Palace,
Mahatma Gandhi Marg, Lucknow 226 001, India

 

Bael (Aegle marmelos (L.) Corr.) is an important medicinal plant of India. Leaves, fruits, stem and roots of
A. marmelos have been used in ethno medicine to exploit its’ medicinal properties including astringent, antidiarrheal antidysenteric, demulcent, antipyretic and anti-inflammatory activities. Compounds purified from bael have been proven to be biologically active against several major diseases including cancer, diabetes and cardiovascular diseases. Preclinical studies indicate the therapeutic potential of crude extracts of A. marmelos in the treatment of many microbial diseases, diabetes and gastric ulcer. This review covers the biological activities of some isolated chemical constituents of A.marmelos and preclinical studies on some crude extracts and pure compounds to explore novel bioactive compounds for therapeutic application.

 

Papers

 

Indian Journal of Experimental Biology

Vol. 47, November 2009, pp.862-870

 

 

Establishment of CHO cell line expressing human MCHR2 gene and research of its molecular characteristics

Chengfu Yuan1,2, Youquan Bu1, Joyeeta Sengupta1, Junxia Yang1, Xiuning Huang1, Li Cheng1, Qin Zhang1, Faping Yi1,
Geli Liu1 & Fangzhou Song1,*

1Department of Biochemistry & Molecular Biology, Molecular Medicine & Cancer Research Center,

Chongqing Medical University, YiXueYuan Road No. 1, Chongqing 400016, China

2Department of Biochemistry, Medical School of HuBei Institute for Nationalities, EnShi 445000, China

Received 22 May 2009; revised 31 July 2009

The whole length of MCHR2 gene cDNA fragment was amplified by PCR using human fetal brain cDNA library as template. The pcDNA3.1 (+)/MCHR2 eukaryotic expression vector was constructed successfully. The recombinant pcDNA3.1 (+)/MCHR2 plasmid was transfected into Chinese hamster ovary (CHO) cell by lipofectamineTM2000, after G418 selection and then the CHO cell line expressing MCHR2 gene was established. The MCHR2 gene expression was tested by RT-PCR, western blotting and immunofluorescence. The maximum binding (Bmax) of CHO cell line was 309.97±1.14 fM·mg-1protein and the dissociation constant (Kd value) was 0.170±0.0006 nM. MCH could stimulate Ca2+ release, its 50% effective concentration (EC50) was 2.32±0.01 nM. The construction of the CHO cell line and the research of MCHR2 molecular characteristics have established a good experimental basis for the further research about the function of MCHR2 gene.

 

 

Indian Journal of Experimental Biology

Vol. 47, November 2009, pp.871-879

 

 

Evaluation of aroA deletion mutant of Salmonella enterica subspecies enterica serovar Abortusequi for its vaccine candidate potential

Javed Alam, B R Singh*, D Hansda, V P Singh & J C Verma

National Salmonella Centre (Vet.), Division of Bacteriology and Mycology
Indian Veterinary Research Institute (IVRI), Izatnagar 243122, India

Received 26 February 2009; revised 20 July 2009

The present study on a defined deletion aroA mutant (B-26) of Salmonella enterica subspecies enterica serovar Abortusequi (S. Abortusequi) for residual virulence and safety in experimental model revealed that the virulence of the strain was at no difference in any of the cell assays (caprine alveolar macrophages, bovine alveolar macrophages, guinea pig blood mononuclear cells and horse blood mononuclear cells) than that of its parent virulence plasmid cured (S-787) and wild type (E-156) strains. The mutant did not cause any apparent illness in baby guinea pigs (15 days old), adult male and female guinea pigs and also not in pregnant (54-55 days of gestation) guinea pigs through oral (4.2 × 109 cfu/ animal) and intramuscular (im) routes (4.2 × 107 cfu/ animal). In pregnant females the mutant also induced abortion as its parent (E-156) though to lesser extent (33%) than the parent strain (100%) on inoculation through intravaginal (4.2 × 109 cfu/ animal) and intraperitoneal (4.2 × 107 cfu/ animal) routes. The babies born from mutant inoculated mothers survived better and were also resistant to intraperitoneal lethal challenge (7.82 × 109 cfu/ animal) with 100% protection. Female guinea pigs challenged after 135-165 days of inoculation with the mutant afforded 100% protection from abortion and mortality caused by lethal infection (7.82 × 109 cfu/ animal) of wild type S. enterica Abortusequi (E-156). The study revealed that aroA mutant (B-26) was safe through oral and im routes for immunization and afforded 100% protection against salmonellosis for more than 5.5 months in guinea pigs. Although immunization with aroA mutant in experimental model afforded good protection against abortion and mortality induced by S. Abortusequi, further studies are needed in horses to exploit the strain’s vaccine potential in the natural host.

 

 

 

Indian Journal of Experimental Biology

Vol. 47, November 2009, pp.880-892

 

 

 

Neuroprotective effect of MK-801 against intra-striatal quinolinic acid induced behavioral, oxidative stress and cellular alterations in rats

Harikesh Kalonia1, Puneet Kumar1, Bimla Nehru2 & Anil Kumar1,*

1Pharmacology Division, University Institute of Pharmaceutical Sciences,

UGC Centre for Advanced Studies, Panjab University, Chandigarh 160 014, India

2 Department of Biophysics, Panjab University, Chandigarh 160014, India

Received 22 February 2009; revised 22 June 2009

Huntington’s Disease (HD) is a common neurodegenerative disorder characterized by motor disturbances, subcortical dementia and psychiatric disturbances. Pathogenesis of HD revolves so far around excitatory amino acids as the primary cause of neuronal loss. However, number of recent reports suggests the involvement of excitotoxicity and oxidative damage. In the present study, first the dose of quinolinic acid that mimics the symptoms of HD was standardized and then the neuroprotective effect of MK-801 (noncompetitive NMDAr antagonist) was evaluated against intrastriatal quinolinic acid induced behavioral, oxidative stress and cellular alterations in rats. A single unilateral (ipsilateral striatum) injections of quinolinic acid (100, 200 and 300 nM) were made in to striatum. Animals were tested for motor functions using actophotometer and rotarod apparatus. Quinolinic acid (300 nM) significantly reduced the body weight and caused motor in-coordination and produced oxidative damage in the cortex and striatum as indicated by raised lipid peroxidation, nitrite concentration, depletion of superoxide dismutase, catalase and different glutathione levels. Beside, quinolinic acid (300 nM) significantly altered the mitochondrial enzymes complex levels and caused histopathological alterations in the striatum. MK-801(0.02, 0.04, 0.08 mg/kg, ip) treatment significantly improved body weight, behavioral alterations (locomotor activity and rotarod performance) and attenuated oxidative damage and mitochondrial enzymes complex dysfunction. Besides, MK-801 treatment significantly reversed histopathological alterations in striatum. The results suggest antioxidant and neuroprotective action of MK-801 against the quinolinic acid induced Huntington’s like behavioral, oxidative stress and cellular alterations in rats.

 

 

 

Indian Journal of Experimental Biology

Vol. 47, November 2009, pp.893-899

 

 

Chronic prenatal restraint stress induced memory impairment in passive
avoidance task in post weaned male and female Wistar rats

Saju Binu Cherian1, K L Bairy2 & Muddanna S Rao3

1Department of Anatomy, Melaka Manipal Medical College, Manipal Campus, ICHS, Manipal, Karnataka, 576 104, India
2Department of Pharmacology, and 3Department of Anatomy, Kasturba Medical College, Manipal, 576 104, India

Received 22 February 2008; revised 19 August 2009

With a view to examine the effect of chronic maternal stress on cognitive function in the offspring during young age, pregnant Wistar rats were subjected to restraint stress from embryonic day 11 till delivery. Male and female pups born to these stressed rats were subjected to passive avoidance test on postnatal day 30 and 31. Results were compared with rats of the same age and sex born to control mothers, which were not stressed. The results showed that prenatal maternal restraint stress impairs the memory retention during young age in both sexes. The memory retention deficit induced by maternal restraint stress was evident in the decreased latency to enter the dark compartment of passive avoidance apparatus by the rats born to stressed mothers. The observed behavioral deficit may be due to the insult of stress on the developing hippocampus, a structure of the brain concerned with learning and memory. The results suggest that prolonged prenatal stress leads to long lasting malfunction in the behavioral development during young age in both male and female young rats. However when compared to their respective stress naïve controls, it seems evident that prenatal restraint stress has a less effect on females which could be due to their oesterogenic effects. These data reinforce the view that prenatal stress affects cognitive development in a sex-specific manner.

 

 

Indian Journal of Experimental Biology

Vol. 47, November 2009, pp.900-905

 

 

 

Tamarindus indica L. and Moringa oleifera M. extract administration
ameliorates fluoride toxicity in rabbits

R Ranjan1*, D Swarup1, R C Patra1 & Vikas Chandra2

Division of Medicine1 and Physiology and Climatology2 Indian Veterinary Research Institute, Izatnagar 243 122, India

Received 13 March 2009; revised 16 July 2009

Aqueous extracts of T. indica fruit pulp (100 mg/ kg body weight) and M. oleifera seeds (50 mg/ kg body weight) orally once daily for 90 days lowered plasma fluoride concentrations in rabbits receiving fluorinated drinking water (200 mg NaF/ Liter water). Cortical indices and metaphysial width in animals receiving extracts also revealed beneficial effects of plant extracts. Changes in plasma biochemistry suggested less hepatic and renal damages in animals receiving plant extracts along with fluorinated water in comparison to that receiving fluorinated water alone. Preliminary results revealed these plant extracts have some potential to mitigate fluoride toxicity.

 

 

 

Indian Journal of Experimental Biology

Vol. 47, November 2009, pp.906-910

 

 

Solar and artificial ultraviolet-B induced erythrocytes
hemolysis with photosensitizers

Sunil Kumar*, Shoma Devi, Prashasti Misra & Priyanka

Environmental Toxicology Laboratory, Department of Zoology, D AV (P G) College, Dehradun 248 001, India

Received 27 January 2009; revised 5 August 2009

Aim of this study was to monitor the solar ultraviolet-B intensity and to compare the phototoxic effect of different intensity of natural and artificial ultraviolet-B on human red blood cells in presence of compounds as riboflavin and chloroquine. Photohemolysis of erythrocytes was studied under natural solar radiation and artificial ultraviolet-B radiation of 312 nm. Monitoring of solar ultraviolet-B radiation was performed in Garhwal region of Uttarakhand, India. Level of solar ultraviolet-B measured show seasonal and altitudinal variations. Monthly average of solar UV-B intensity was minimum in the month of December and January (0.299 mw/cm2) and maximum in the month of July and August (1.027 mw/cm2). Natural solar radiation intensities 0.402 mw/cm2 and 0.824 mw/cm2 of the month of January and June were used in the photohemolysis experiment. Two intensities of artificial UV-B i.e. 0.824 mw/cm2 and a double intensity 1.65 mw/cm2 were also used. Results on human erythrocytes hemolysis indicate that haemolysis was highest i.e. 71% in chloroquine + artificial ultraviolet-B intensity (1.65 mw/cm2) followed by 62% in chloroquine + artificial ultraviolet-B (0.824 mw/cm2) exposed groups and 54% in natural solar radiation intensity 0.824 mw/cm2 + chloroquine. Natural solar UV-B alone caused 17% hemolysis and show dose response relationship.  A difference in phototoxicity was observed in natural solar and artificial UV-B of same intensity. Artificial UV-B was found more toxic. Riboflavin was more phototoxic in presence of solar light, while chloroquine was more phototoxic with artificial UV-B.

 

Indian Journal of Experimental Biology

Vol. 47, November 2009, pp.911-915

 

 

Quantitative PCR: A quality control assay for estimation of viable virus content in live attenuated goat pox vaccine

D J Kallesh1, M Hosamani2*, V Balamurugan1, V Bhanuprakash1, V Yadav1 & R K Singh1

1 Division of Virology, Indian Veterinary Research Institute, Mukteswar 263 138, Nainital, India

2. Indian Veterinary Research Institute, Hebbal Campus, Bangalore 560 024, India

Received 26 August 2008; revised 29 July 2009

Efficacy of live viral vaccine and vaccine-induced sero-conversion depends on the optimum number of live virus particles in a vaccine dose, which is one of the important aspects of quality control. In the present study, TaqMan® probe quantitative polymerase chain reaction (QPCR) based on conserved DNA pol gene of capripoxvirus was developed for the quality control of attenuated monovalent goatpox and/or combined attenuated goatpox and peste des petits ruminants (PPR) vaccines. Cells infected with vaccine virus were harvested at critical time point and subjected to QPCR. A critical time point for harvest of Vero cells infected with various log10 dilutions of reference virus was determined to be 36 h (highest slope 3.062), by comparison of slopes of standard curves established with harvests at different time intervals. The assay method was evaluated using different batches of goatpox vaccine, and bivalent goatpox and PPR vaccine. The titers estimated by QPCR and TCID50 method were comparable to each other. The QPCR assay thus, could be used as an alternate method or supplementary tool for estimation of live GTPV particles in monovalent goatpox or bivalent goatpox and PPR vaccines.

 

 

Indian Journal of Experimental Biology

Vol. 47, November 2009, pp.916-920

 

 

Genetic engineering of avian pathogenic E. coli to study the functions
of FimH adhesin

H H Musa1,2§, S F He1§, S L Wu1, C H Zhu1, Z H Liu1, Z N Zhang1, V S Raj3, R X Gu 4 & G Q Zhu1*

1College of Veterinary Medicine, Yangzhou University, Yangzhou 225009, China

2Faculty of Veterinary Science, University of Nyala, Nyala 155, Sudan

3Ranbaxy Laboratories Limited Research & Development Center, Gurgaon 122015, India

4Institute of Dairy Science and Technology, Yangzhou University, Yangzhou 225009, China

 

Received 13 February 2009; revised 22 July 2009

Adhesion of pathogen to host cells is an important prerequisite for successful colonization and establishment of the pathogenesis. The aim of this study is to examine the function of FimH adhesin in the adherence of avian pathogenic E. coli to porcine intestinal epithelial cell lines (IPEC-J2) and human lung epithelial cell line (A549) in an in vitro infection model. Three strains of avian pathogenic Escherichia coli (APEC) and one strain of non-pathogenic E coli were used. The isogenic FimH mutants were constructed by λ Red-mediated recombination system. The wild types and mutants strains were adhered to the host cells with different adherence patterns in certain incubation time. The results demonstrated that the adherence of the isogenic FimH mutants to the porcine intestinal epithelial cells (IPEC-J2) were similar to those of wild types. However, the adherences of isogenic FimH mutants to human lung epithelial cells (A549) were significantly different from the wild types. A549 cell can be used as a type of cell model for colonization of the chicken extraintestinal. FimH offers a unique opportunity to investigate the role of the strength of adhesion independently from the many other factors that may affect surface colonization.

 

Notes and News

Indian Journal of Experimental Biology

Vol. 47, November 2009, pp.921-924

 

 

Protocol for improved extraction and PCR amplification of genomic DNA
from liverwort, Plagiochasma appendiculatum

Arvind Soni & Anil Kumar*

Plant Genomic Lab, Molecular Biology and
Genetic Engineering Division,
National Botanical Research Institute, Luckonw 266 001, India

Received 9 February 2009; revised 22 June 2009

A simplest method was followed for isolation of high quality genomic DNA form thallus of Plagiochasma appendiculatum that contained large quantities of polyphenols, terpenoids, tannins, contamination of high amount of RNA and polysaccharides. The method involved a modification of CTAB procedure using PVP (1%) and LiCl (4M) solution to remove polyphenols and RNA and some other binding proteins. The present protocol was found suitable for restriction enzyme digestion and random amplified polymorphic DNA (RAPD) analysis.

 

 

Indian Journal of Experimental Biology

Vol. 47, November 2009, pp.925-928

 

 

Assessment of genetic fidelity of micropropagated apple rootstock plants, EMLA 111, using RAPD markers

R Gupta, M Modgil* & S K Chakrabarti1

Department of Biotechnology, Dr Y S Parmar University of Horticulture & Forestry, Nauni, Solan 173230, India

1Division of Crop Improvement, Central Potato Research Institute, Shimla, India

Received 23 December 2008; revised 16 July 2009

EMLA 111(East Malling Long Ashton), a clonal rootstock of apple (Malus pumila Mill.) was micropropagated using axillary bud and shoot apices. The present work was carried out to assess the genetic fidelity of micropropagated plants using random amplified polymorphic DNA (RAPD). Ten arbitrary decamer primers have been used to amplify genomic DNA from in vitro raised field material and mother plant. A total of 57 amplified products were obtained, out of which 53 were monomorphic across the mother tree and its tissue culture raised progenies. Of the ten primers used, 8 showed RAPD profiles which were identical to the mother plant. Similarity matrix based on Jaccard’s coefficient revealed that pairwise value between the mother plant and its tissue cultured plants ranged from 0.93 to 1.00 and among tissue cultured plants, it was 0.92 to 1.00, thus indicating a high degree of genetic fidelity.

 

Indian Journal of Experimental Biology

Vol. 47, November 2009, pp.929-932

 

 

Influenza A (H1N1) pandemic: Preparedness and clinical management

Madhu Khanna & Neha Gupta

 

Department of Respiratory Virology

VP Chest Institute, University of Delhi, Delhi 110 007, India