Indian Journal of Experimental Biology

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VOLUME 49

NUMBER 2

FEBRUARY 2011

CODEN: IJEB (A6) 49 (2) - (2010) 77-162

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

 

CONTENTS

 

Review Article

 

Therapeutic effects of proanthocyanidins on the pathogenesis of periodontitis — An overview

83

      Jayamathi Govindaraj, Pamela Emmadi & Rengarajulu Puvanakrishnan

 

 

 

Papers

 

Lentiviral-mediated RNAi targeting p38MAPK ameliorates high glucose-induced apoptosis in osteoblast MC3T3-E1 cell line

94

      Zhengping Feng, Huacong Deng, Jia Du, Danyan Chen, Rong Jiang &
Xiaoyan Liang

 

 

 

Silencing of Bmi-1 gene by RNA interference enhances sensitivity to doxorubicin in breast cancer cells

105

      Xiangmei Wu, Xing Liu, Joyeeta Sengupta, Youquan Bu, Faping Yi,
Changdong Wang, Yanyan Shi, Yong Zhu, Qingfang Jiao & Fangzhou Song

 

 

 

Human sodium iodide symporter (hNIS) in fibroadenoma breast—A immunohistochemical study

113

      Ruchi Rai, Ashutosh Shrivastava, Ashwani Tandon, Madan M Godbole,
Sandeep Kumar, Vinita Das, Varsha Dwivedi & Lily Pal

 

 

 

Proteomic analysis of cerebral synaptosomes isolated from rat model of Alzheimer’s disease

118

      Huiyun Yang, Haixuan Qiao & Xin Tian

 

 

 

Antioxidant effect of Lagerstroemia speciosa Pers (Banaba) leaf extract in streptozotocin-induced diabetic mice

125

      Saumya S M & P Mahaboob Basha

 

 

 

Ameliorative role of atorvastatin on methionine-induced hyperhomocysteinemia and hematological changes in albino rats

132

      Uma Bhandari, Rahila Ahmad Pathan, Vinay Kumar & Naresh Khanna

 

 

 

Development and in vitro characterization of a bivalent DNA containing HN and F genes of velogenic Newcastle disease virus

140

      Uttara Chaturvedi, Shahina Kalim, G Desai, Barkha Ratta, Rajiv Kumar,
P V Ravindra, Sudesh Kumar, B B Dash, Sangeeta Tiwari, A P Sahoo &
Ashok K Tiwari

 

 

 

Acute and sub-acute toxicity of ethanolic extract of Canthium mannii Hiern stem bark on
Mus musculus

146

      J Wabo Pone, Mpoame Mbida & C F Bilong Bilong

 

 

 

Induction of systemic resistance in different varieties of Solanum tuberosum by pure and crude elicitor treatment

151

      Himanshu S Bariya, Vasudev R Thakkar, Amit N Thakkar & R B Subramanian

 

 

 

—————————

Announcement

 

National Symposium on Microwave Field Measurement, Biological Effects and
Application in Nano-Science (MICRO-NANO-2011)

 

4 and 5 March 2011, JNU, New Delhi

 

Organized and sponsored by the Jawaharlal Nehru University, New Delhi and Microwave Applications Society of India (MASI), New Delhi in collaboration with IEEE-MTT Society, the symposium will be based on nanotechnology, biomedical, biophysical and environmental research issues.

The topics to be covered will include but not limited to the following: (i) Principle of microwave radiations (ii) Electromagnetic fields-bio-interactions and solution
(iii) Electromagnetic fields and environment (iv) Radiofrequency fields measurement and dosimetry (v) Radiation hazards and safety standards (vi) Nanotechnology in wastewater treatment, and (vii) Nanoscience applications in medical, biological and environmental issues.

For further details, kindly contact Prof. J. Behari, School of Environmental Sciences, Jawaharlal Nehru University, New Delhi 110 067. Telephone: +91-011-2670 4323 (O),
+91-011-2617 5857 (R); Fax: +91-011-2616 5856; E-mail: masijnudelhi@gmail.com

 

————————

Editor’s Note

 

The Indian Journal of Experimental Biology is covered by the following international abstracting and indexing services:

 

Science Citation Index ExpandedTM

PubMed (http://www.ncbi.nim.nih.gov/)

MEDLINE

BIOSIS

Chemical Abstracts Service

Excerpta Medica

Informascience

Refrativnyi Zhurnal

Zoological Records

 

Author Index

Bariya Himanshu S

151

Basha P Mahaboob

125

Bhandari Uma

132

Bilong Bilong C F

146

Bu Youquan

105

 

 

Chaturvedi Uttara

140

Chen Danyan

94

 

 

Das Vinita

113

Dash B B

140

Deng Huacong

94

Desai G

140

Du Jia

94

Dwivedi Varsha

113

 

 

Emmadi Pamela

83

 

 

Feng Zhengping

94

 

 

Godbole Madan M

113

Govindaraj Jayamathi

83

 

 

Jiang Rong

94

Jiao Qingfang

105

 

 

Kalim Shahina

140

Khanna Naresh

132

Kumar Rajiv

140

Kumar Sandeep

113

Kumar Sudesh

140

Kumar Vinay

132

 

 

Liang Xiaoyan

94

Liu Xing

105

 

 

Mbida Mpoame

146

 

 

Pal Lily

113

Pathan Rahila Ahmad

132

Pone J Wabo

146

Puvanakrishnan Rengarajulu

83

 

 

Qiao Haixuan

118

 

 

Rai Ruchi

113

Ratta Barkha

140

Ravindra P V

140

Sahoo A P

140

Saumya S M

125

Sengupta Joyeeta

105

Shi Yanyan

105

Shrivastava Ashutosh

113

Song Fangzhou

105

Subramanian R B

151

 

 

Tandon Ashwani

113

Thakkar Amit N

151

Thakkar Vasudev R

151

Tian Xin

118

Tiwari Ashok K

140

Tiwari Sangeeta

140

 

 

Wang Changdong

105

Wu Xiangmei

105

 

 

Yang Huiyun

118

 

 

Yi Faping

105

 

 

Zhu Yong

105

 

Keyword Index

Antioxidant property

83

Alzheimer’s disease

118

Apoptosis

94

Atorvastatin

132

 

 

Bicistronic gene construct

140

Bmi-1

105

Breast cancer

105

 

 

Canthium mannii

146

 

 

Defense related enzymes

151

2-DGel electrophoresis

118

Diabetes

125

DNA vaccine

140

Doxorubicin

105

 

 

Elicitor

151

 

 

Fibroadenoma breast

113

Free radicals

125

 

 

High glucose

94

Human sodium iodide
symporter (hNIS)

 

113

Hyperhomocysteinemia

132

Hypersensitivity response

151

 

 

Immunohistochemistry

113

In vitro expression

140

 

 

Lagerstroemia

125

Lentivirus

94

Lipid profile

132

 

 

MC3T3-E1 cell line

94

Methionine

132

 

 

NDV

140

Nitric oxide

83

Oxidative stress

125

 

 

P38MAPK

94

Periodontitis

83

Phytophthora infestans

151

Proanthocyanidin

83

Proteomics

118

 

Reactive oxygen species

83

RNA interference

94,105

 

 

Solanum tuberosum

151

 

 

Stem-bark extract

146

Synaptosomes

118

 

 

Toxicity

146

 

 

Indian Journal of Experimental Biology

Vol. 49, February 2011, pp. 83-93

 

 

Review Article

 

 

 

Therapeutic effects of proanthocyanidins on the pathogenesis of periodontitis
An overview

Jayamathi Govindaraj1, Pamela Emmadi2 & Rengarajulu Puvanakrishnan3*

Department of 1Biochemistry and 2Periodontics, Meenakshi Ammal Dental College, Alapakkam Main Road,
Maduravoyal, Chennai 600 095, India
3Department of Biotechnology, Central Leather Research Institute (CLRI), CSIR, Chennai 600 020, India

 

Periodontitis is a bacterially induced chronic inflammatory disease that destroys the connective tissue and bone that support teeth. Bacteria initiates periodontitis and destruction of the alveolar bone and periodontal connective tissue is clearly observed. But, the events occuring between these two points of time remain obscure and this study focusses on these aspects. The proanthocyanidins (PC) have variable pharmacological and nutraceutical benefits including improvement of ischemic cardiovascular disease, prevention of atherosclerosis and antiarthritic, anticancer and antimicrobial activities. The benefits associated with the antioxidant activity of PC have been evaluated both in vivo and in vitro. But, reports on the ameliorative effects of PC on oral diseases and specifically on periodontitis are very few. Hence, a novel attempt is made to review the possible protective effects of PC and its mechanism of action in periodontitis and also to show whether PC could be developed as a therapeutic agent for periodontitis.

 

 

 

Papers

Indian Journal of Experimental Biology

Vol. 49, February 2011, pp. 94-104

 

 

Lentiviral-mediated RNAi targeting p38MAPK ameliorates high glucose-induced apoptosis in osteoblast MC3T3-E1 cell line*

Zhengping Feng1, Huacong Deng1, Jia Du2, Danyan Chen1, Rong Jiang2 & Xiaoyan Liang2

1Department of Endocrinology, The First Affiliated Hospital, Chongqing Medical University, Chongqing, China, 400016

2 Laboratory of Stem Cell and Tissue Engineering, Chongqing Medical University, Chongqin, China, 400016

Received 5 April 2010; revised 12 October 2010

The p38 mitogen activated protein kinase (p38MAPK) pathway is an important signaling cascade involved in cell growth, differentiation and apoptosis. High glucose activates p38MAPK pathway in different cells, including osteoblasts. In the present study, role of p38MAPK in high glucose induced osteoblast apoptosis and potential of RNA interference (RNAi) targeting p38MAPK as a therapy strategy have been reported. Lentiviral-mediated RNAi effectively reduced p38MAPK and p-p38MAPK expressions in osteoblastic cell line (MC3T3-E1) following high glucose (22 mM) induction. Inhibition of p38MAPK activity significantly suppressed high glucose induced apoptosis of MC3T3-E1 cell and was confirmed by flow cytometry and ultra-structural examination by transmission electronic microscope. Inhibition of p38MAPK also significantly attenuates caspase-3 and bax protein expressions, but increased significantly bcl-2 expression as determined by Western blot analysis. The results suggested that p38MAPK mediates high glucose induced osteoblast apoptosis, partly through modulating the expressions of caspase-3, bax and bcl-2. Inhibition of p38MAPK with lentiviral-mediated RNAi or its specific inhibitor provides a new strategy to treat high glucose induced osteoblast apoptosis.

 

 

 

Indian Journal of Experimental Biology

Vol. 49, February 2011, pp. 105-112

 

 

 

Silencing of Bmi-1 gene by RNA interference enhances sensitivity to doxorubicin in breast cancer cells

Xiangmei Wu1,2, Xing Liu3, Joyeeta Sengupta2, Youquan Bu2, Faping Yi2, Changdong Wang2, Yanyan Shi2,
Yong Zhu2, Qingfang Jiao2 & Fangzhou Song2*

1 Department of Physiology, Chongqing Medical University, Chongqing 400016, PR China

2 Molecular Medicine and Cancer Research Center, Chongqing Medical University, Chongqing 400016, PR China

3 Department of Pediatric Urology, Chongqing Children’s Hospital, Chongqing Medical University, Chongqing 400014, PR China

Received 8 February 2010; revised 18 November 2010

The oncogene Bmi-1 is highly up-regulated in breast carcinoma and is found to be efficient in preventing apoptosis of the cancer cells. Doxorubicin is an important chemotherapeutic agent against breast carcinoma. However, the effective therapeutic response to doxorubicin is often associated with severe toxicity. The present study is targetted at developing a strategy to increase doxorubicin sensitivity to lower doses without compromising its efficacy. A stable cell line with a persistent silencing of Bmi-1 was established. MTT assay was performed to evaluate 50% inhibitory concentration (IC50) values of doxorubicin. Apoptosis was detected by FCM and the expression of related genes [phosphor-Akt (pAkt), totle-Akt (tAkt), Bcl-2 and Bax] was studied by Western blot. In vivo, the sensitivity of the tumor tissues against doxorubicin was evaluated by transplanted MCF-7 nude mice model and the apoptosis of tissue cells was detected by TUNEL assay. The expression of pAkt and Bcl-2 was down-regulated, whereas Bax was up-regulated in Bmi-1 silencing cells. The results obtained indicated that silencing of Bmi-1 can render MCF-7 cells more sensitive to doxorubicin which induced a significantly higher percentage of apoptosis cells in vitro and in vivo. All together these results clearly demonstrate that Bmi-1 siliencing combined treatment of doxorubicin might be a new strategy for biological treatment on breast cancer.

 

 

Indian Journal of Experimental Biology

Vol. 49, February 2011, pp. 113-117

 

 

Human sodium iodide symporter (hNIS) in fibroadenoma breast—A immunohistochemical study

Ruchi Rai1, Ashutosh Shrivastava3, Ashwani Tandon4, Madan M Godbole3, Sandeep Kumar2,*, Vinita Das1, Varsha Dwivedi2 & Lily Pal4

Departments of 1Obstetrics & Gynaecology and 2Surgery, Chhatrapati Shahuji Maharaj Medical University,
Lucknow 226 003, India

and

3Departments of Endocrinology and 4Pathology, Sanjay Gandhi Postgraduate Institute of Medical Sciences,
Lucknow 226 014, India

Received 29 June 2010; revised 12 October 2010

Human sodium iodide symporter (hNIS), responsible for the active transport of iodine is an integral plasma membrane glycoprotein present in the thyroid cells and extrathyroid tissues like breast and salivary glands. If its functional form is unequivocally shown in benign or malignant breast tissues, then it may serve as a basis for diagnosis and treatment using radioactive iodine. With an aim to analyze the hNIS expression in a distinct benign breast condition of fibroadenoma, biopsy proven fibroadenoma tissues, normal non-lactating breast tissue and biopsy proven infiltrating duct carcinoma tissues were examined for hNIS expression using immunohistochemistry. Out of 20 biopsy proven fibroadenoma tissues, 19 (95%) showed positivity for hNIS protein and only one was negative. Of these 10% were mildly positive, 50% cases were moderately positive and 35% showed intense positivity. None of the control tissue obtained from reduction mammoplasty specimens or normal breast tissues samples (5 cms away from the tumor) were positive. hNIS was also intensely positive  in 9 out of 10 (90%) infiltrating duct carcinoma tissues and moderately positive in one case. These preliminary results show that hNIS was present in high frequency as demonstrated by immunohistochemistry in fibroadenoma breast.

 

 

Indian Journal of Experimental Biology

Vol. 49, February 2011, pp. 118-124

 

 

Proteomic analysis of cerebral synaptosomes isolated from rat model of Alzheimer’s disease

Huiyun Yang, Haixuan Qiao & Xin Tian*

Tianjin Medical University, Tianjin, China 300070

Received 14 October 2009; revised 22 October 2010

Alzheimer’s disease (AD) is a common and devastating disease and there is no readily available biomarker to aid diagnosis or monitor progression of it. To further understand the pathogenic mechanism of AD, proteomic approach was used to study the cerebral synaptosomes proteins of rats injected with Aβ1-40. Compared with the untreated samples,
14 proteins were found apparently altered through 2-dimensional gel electrophoresis. 12 of them were down-regulated
and 2 were up-regulated. Three proteins including alpha-2-globin chain, peptidyl-prolycis-trans isomerase A (PPIaseA)
and cofilin-1 protein were
identified by matrix-assisted laser desorption ionization time of flight mass spectrometry (MALDI-TOF-MS) and SWISS-PROT database query. Alpha-2-globin chain has not been shown to be associated with AD. PPIaseA and cofilin-1 protein are correlated with cell apoptosis and signaling. The altered proteins identified may help to understand the pathogenesis of AD.

 

 

Indian Journal of Experimental Biology

Vol. 49, February 2011, pp. 125-131

 

 

Antioxidant effect of Lagerstroemia speciosa Pers (Banaba) leaf extract in streptozotocin-induced diabetic mice

Saumya S M & P Mahaboob Basha*

Department of Zoology, Bangalore University, Bangalore 560 056, India

Received 26 November 2009; revised 21 October 2010

Aqueous leaf extract of L. speciosa (banaba) effectively decreased the blood glucose in streptozotocin-induced diabetic mice after 15th day of banaba exposure. Further, banaba leaf extract have the potential to inhibit lipid peroxidation
and effectively intercept/neutralize reactive oxygen species such as super oxide, H2O2 and NO based free radicals.
The aqueous banaba leaf extract (150 mg/kg bodyweight) duly reduced STZ generated reactive intermediates and radical species helping to regulate normal levels of antioxidative markers like superoxide dismutase, catalase, glutathione-
S-transferase and reduced glutathione.

 

Indian Journal of Experimental Biology

Vol. 49, February 2011, pp. 132-139

 

 

Ameliorative role of atorvastatin on methionine-induced hyperhomocysteinemia and hematological changes in albino rats

Uma Bhandari1,* Rahila Ahmad Pathan1, Vinay Kumar1 & Naresh Khanna2

1Department of Pharmacology, Faculty of Pharmacy, Hamdard University, New Delhi 110 062, India

2Department of Pharmacology, University College of Medical Sciences & Guru Teg Bahadur Hospital, Delhi 110 095, India

Received 8 February 2010; revised 18 October 2010

Methionine (1g/kg, po) administration to pathogenic control rats for 30 days significantly increased the levels of homocysteine, total cholesterol (TC), low density lipoprotein (LDL-C), very low density lipoprotein (VLDL-C) and triglycerides (TGs) and decreased the levels of high density lipoprotein (HDL-C) in serum. Hematological observations of the peripheral blood smears of pathogenic rats fed with methionine also showed crenation of RBCs cell membrane and significant increase in total leukocyte count, differential leukocyte count and platelet counts with significant decrease in the mean hemoglobin levels as compared to vehicle control rats. Administration of atorvastatin (0.2 mg/kg/po) to hyperhomocysteinemic rats significantly decreased the levels of homocysteine, TC, TGs, LDL-C and VLDL-C and increased the levels of HDL-C in serum. The present results provide clear evidence that oral treatment with atorvastatin exhibit homocysteine and lipid lowering activity and also reversal of hematological changes induced by methionine
in albino rats.

 

 

Indian Journal of Experimental Biology

Vol. 49, February 2011, pp. 140-145

 

 

Development and in vitro characterization of a bivalent DNA containing HN and F genes of velogenic Newcastle disease virus

Uttara Chaturvedi, Shahina Kalim1, G Desai2, Barkha Ratta, Rajiv Kumar, P V Ravindra, Sudesh Kumar, B B Dash3, Sangeeta Tiwari4, A P Sahoo & Ashok K Tiwari*

Molecular Biology Laboratory, Division of Animal Biotechnology

Indian Veterinary Research Institute, Izatnagar 243 122, India

Received 1 June 2010; revised 26 October 2010

Newcastle disease (ND) is highly contagious, economically important viral disease affecting most of avian species worldwide. Newcastle disease virus (NDV) has single stranded negative sense RNA genome which encodes for six structural and two non-structural proteins. Envelope glycoproteins i.e. hemagglutinin-neuraminidase (HN) and the fusion (F), elicit protective immune response. In this study, HN and F genes of velogenic (virulent) strain were amplified and cloned at multiple cloning sites A and B, respectively into pIRES bicistronic vector for use as bivalent DNA vaccine against ND. The recombinant plasmid was characterized for its orientation by restriction enzyme digestion and PCR. Expression of HN and F genes was assessed in transfected Vero cells at RNA level using RT-PCR in total RNA as well as protein level using IFAT, IPT and western blot using NDV specific antiserum. All these experiments confirmed that HN and F genes cloned in recombinant pIRES.nd.hn.f are functionally active. The recombinant construct is being evaluated as DNA vaccine against ND.

 

 

Indian Journal of Experimental Biology

Vol. 49, February 2011, pp. 146-150

 

 

Acute and sub-acute toxicity of ethanolic extract of Canthium mannii Hiern
stem bark on Mus musculus

J Wabo Poné* & Mpoame Mbida

Department of Animal Biology, Laboratory of Applied Biology and Ecology, Faculty of Science,
University of Dschang, PO Box 067 Dschang, Cameroon.

and

C F Bilong Bilong

Department of Animal Biology and Physiology, Laboratory of General Biology, Faculty of Science,
University of Yaoundé 1, PO Box 812 Yaoundé, Cameroon.

Received 23 July 2009, revised 1 November 2010

Acute and sub-acute toxicity of ethanolic extract (ETE) of C. mannii was assessed on white mice (Mus musculus). After 48 h of extract administration, no death was registered. It was deduced that the LD50 was indisputably higher than 16 g/kg body weight. The sub-acute toxicity test was based on the daily administration of three doses of ETE (300, 600 and 1200 mg/kg body weight) for four weeks; 1% DMSO served as negative control. As for the first experiment, no sign of toxicity was registered. Conversely, the sub acute doses stimulated and increased the weight-rate of mice after 7 days of treatment. Except for the spleen weight, the doses administrated did not modify the weight index. It was observed that, sub-acute doses induced and increased (a) the food (particularly) and water consumption according to time and (b) the number of red and white blood cells. It was thought that, ETE can stimulate the haematopoietic function. Finally, no time variation of the activity of alanine aminotransferase and aspartate aminotransferase enzyme was observed in the serum of euthanized mice. The results showed the innocuity of ETE of C. mannii and thus validated his utilization in cameroonian traditional pharmacopoea.

 

 

Indian Journal of Experimental Biology

Vol. 49, February 2011, pp. 151-162

 

 

Induction of systemic resistance in different varieties of Solanum tuberosum by pure and crude elicitor treatment

Himanshu S Bariya, Vasudev R Thakkar*, Amit N Thakkar & R B Subramanian

B & R Doshi School of Biosciences, Sardar Patel Maidan, Sardar Patel University,
Vadtal Road, Bakrol, Vallabh Vidyanagar 388 120, India

Received 28 January 2010; revised 13 July 2010

A 10 kD elicitor protein (infestin) produced by Phytopthora infestans was purified and its efficacy for induction of systemic resistance in resistant and susceptible varieties of Solanum tuberosum was studied. Culture filtrates from
P. infestans with and without purified elicitor (infestin) were used as elicitors to understand the effect of purified elicitor (infestin) on development of systemic resistance. Culture filtrate and purified elicitor (infestin) were found to induce hypersensitive reaction on the leaves of resistant varieties, but not on susceptible varieties after 48 h. Culture filtrate devoid of purified elicitor (infestin) did not induce any necrotic spots even on resistant variety. Purified elicitor (infestin) was found to induce glucose oxidase, NADPH oxidase, superoxide dismutase, glutathione reductase, catalase and peroxidase enzymes in resistant S. tuberosum plants, however the induction of these enzymes was low in susceptible varieties. The oxidative enzymes were found to induce earlier than antioxidative enzymes and there was negative correlation between these two groups of enzymes. Levels of salicylic acid, phenylalanine ammonia lyase (PAL), β-1, 3 glucanase and chitinase activities were also found higher in resistant than in susceptible varieties. It was observed that purified elicitor (infestin) was superior to crude culture filtrate, but was not capable of inducing systemic resistance in susceptible varieties.