Indian Journal of Experimental Biology

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VOLUME 52

NUMBER 1

JANUARY 2014

CODEN: IJEB (A6) 52 (1) 1-98 (2014)

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

CONTENTS

 

Review Article

 

 

 

Stud male-originating chemosignals: A luteotrophic agent

5

 

 

G Archunan

 

 

 

Papers

 

 

 

Toll like receptor 2 and CC chemokine receptor 5 cluster in the lipid raft enhances the susceptibility of Leishmania donovani infection in macrophages

17

 

 

Suchandra Bhattacharyya Majumdar, Parna Bhattacharya, Surajit Bhattacharjee, Saikat Majumder, Sayantan Banerjee & Subrata Majumdar

 

 

 

Response of male mice to odours of female mice in different stages of oestrous cycle:
Self-grooming behaviour and the effect of castration

30

 

 

Shanmugam Achiraman, Devaraj SankarGanesh, Soundarapandian Kannan, Soundararajan Kamalakkannan, Natarajan Nirmala & Govindaraju Archunan

 

 

 

Hypolipidemic and antioxidant activity of ethanolic extract of Symplocos racemosa in hyperlipidemic rats: An evidence of participation of oxidative stress in hyperlipidemia

36

 

 

A M Durkar, R R Patil, S R Naik

 

 

 

Antidiabetic activity of a triterpenoid saponin isolated from Momordica cymbalaria Fenzl

46

 

 

Raju Koneri Balwanth, Suman Samaddar & Channakeshava Thimmasandra Ramaiah

 

 

 

Behavioural and neuroendocrine effects of aqueous extract of Boerhaavia diffusa Linn. in mice using tail suspension and forced swim tests – A preliminary study

53

 

 

Dinesh Dhingra & Rekha Valecha

 

 

 

Quercetin and β-sitosterol prevent high fat diet induced dyslipidemia and hepatotoxicity in Swiss albino mice

60

 

 

Kunal Sikder, Nilanjan Das, Swaraj Bandhu Kesh & Sanjit Dey

 

 

Human malaria in C57BL/6J mice: An in vivo model for chemotherapy studies

67

 

 

K Sunita, M Rajyalakshmi, K Kalyan Kumar, M Sowjanya, PVV Satish & D Madhu Prasad

 

 

 

A common HPLC-PDA method for amino acid analysis in insects and plants

73

 

 

M K Dhillon, Sandeep Kumar & G T Gujar

 

 

 

Establishment of an in vitro plantlet regeneration protocol for unique varieties of brinjal (Solanum melongena L.) var. Mattu Gulla and Perampalli Gulla

80

 

 

A Muthusamy, KS Vidya, PK Pratibha, M Radhakrishna Rao, SB Vidhu, KP Guruprasad, U Raghavendra, PM Gopinath & K Satyamoorthy

 

 

 

Potential application of β-1, 3 glucanase from an environmental isolate of
Pseudomonas aeruginosa MCCB 123 in fungal DNA extraction

89

Divya Jose, P Jayesh, Prem Gopinath, A Mohandas & I S Bright Singh

 

 

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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

 

————————

 

Indian Journal of Experimental Biology in Open Access Mode

 

      The Indian Journal of Experimental Biology (IJEB) is now an open access journal in the repository, NISCAIR Online Periodicals Repository (NOPR) [http://nopr.niscair.res.in].

      Full text of all articles published in IJEB from 1999 onwards can now be accessed at NOPR in the open access mode. Papers in the current issue shall be uploaded immediately. Papers published in earlier years shall be added soon.

      NOPR is based on DSpace, a digital repository software, and allows document browsing, document searching and various search options like title, author name, keywords, year, issue, etc.

 

——————————————

 

 

 

 

Author Index

Achiraman Shanmugam

30

Archunan G

5

Archunan Govindaraju

30

 

 

Banerjee Sayantan

17

Bhattacharjee Surajit

17

Bhattacharya Parna

17

Bright Singh I S

89

 

 

Das Nilanjan

60

Dey Sanjit

60

Dhillon M K

73

Dhingra Dinesh

53

Durkar A M

36

 

 

Gopinath PM

80

Gopinath Prem

89

Gujar G T

73

Guruprasad KP

80

 

 

Jayesh P

89

Jose Divya

89

Kalyan Kumar K

67

Kamalakkannan Soundararajan


30

Kannan Soundarapandian

30

Kesh Swaraj Bandhu

60

Koneri Balwanth Raju

46

 

 

Madhu Prasad D

67

Majumdar Subrata

17

Majumdar Suchandra Bhattacharyya


17

Majumder Saikat

17

Mohandas A

89

Muthusamy A

80

 

 

Naik S R

36

Nirmala Natarajan

30

 

 

Patil R R

36

Pratibha PK

80

Raghavendra U

80

Rajyalakshmi M

67

Rao M Radhakrishna

80

 

 

Samaddar Suman

46

Sandeep Kumar

73

SankarGanesh Devaraj

30

Satish PVV

67

Satyamoorthy K

80

Sikder Kunal

60

Sowjanya M

67

Sunita K

67

 

 

Thimmasandra Ramaiah Channakeshava


46

 

 

Valecha Rekha

53

Vidhu SB

80

Vidya KS

80

 

 

Keyword Index

Amino acids

73

Arteether

67

Artesunate

67

 

 

Boerhaavia diffusa

53

 

 

CC chemokine receptor 5

17

Chemosignals

5

 

 

Depression

53

DNA content

80

DNA extraction

89

Dyslipidemia Hepatotoxicity


60

 

 

Eggplant

80

 

 

Flow cytometry

80

Forced swim test

53

Fungus

89

 

 

 

 

Glucose uptake

46

Gradient profile

73

Grooming behaviour

30

 

 

HPLC-PDA

73

Hyperlipidemia

36

Hypolipidemia

36

 

 

Insect

73

Insulin secretion,

46

 

 

Leishmania donovani

17

 

 

Macrophages

17

Momordica cymbalaria

46

Monoamine oxidase

53

Mouse

30

 

 

Oestrus

30

Olfactory luteotrophic memory Pheromone


5

Organogenesis

80

 

 

 

 

Parasitemia

67

Plant

73

Plant-antioxidants

36

Plantlet regeneration

80

Plasmodium falciparum

67

Pseudomonas aeruginosa

89

 

 

Quercetin

60

 

 

Standardization

73

Steatosis

60

Stud male

5

Symplocos racemosa

36

 

 

Tail suspension test

53

Testosterone

30

Tissue culture

80

Toll-like receptor 2

17

 

 

Urine

30

 

 

β-1,3 glucanase

89

β-sitosterol

60

 

 

            Correspondent author is marked by *

 

Indian Journal of Experimental Biology

Vol. 52, January 2014, pp. 5-16

 

 

 

Review Article

 

 

Stud male-originating chemosignals: A luteotrophic agent*

G Archunan

Centre for Pheromone Technology, Department of Animal Science, Bharathidasan University, Tiruchirappalli 620 024, India

The chemosignals from mating male are found to be responsible for protecting his coital partner against pregnancy failure induced by strange male or food-deprivation. The stud male pheromone not only provides luteotrophic support in female of vulnerable condition but also exerts luteotrophic effect in pregnancy-blocked females by inducing pseudopregnancy. The luteotrophic stimulus rendered by stud male to prevent pregnancy failure is mediated through the main olfactory system, and not through the accessory olfactory system, since the accessory olfactory system is primarily involved in perceiving the luteolytic stimulus produced from strange male for causing pregnancy failure. It has been shown that pericopulatory period seems to be crucial in females in the formation of stud male chemosignals, and the olfactory luteotrophic memory of stud male is further proved to be a short-term one. The precise mechanism involved in accessing and retaining the stud male chemical cues is unclear. In this brief review an attempt has been made to bring out the luteotrophic process of stud male chemosignals, the olfactory pathway and critical period to access the signals. The possible neural mechanism and neural chemistry underlying the formation and recognition of mating male chemical cues are also highlighted.

 

Papers

 

Indian Journal of Experimental Biology

Vol. 52, January 2014, pp. 17-29

 

 

Toll like receptor 2 and CC chemokine receptor 5 cluster in the lipid raft enhances the susceptibility of Leishmania donovani infection in macrophages

Suchandra Bhattacharyya Majumdar*, Parna Bhattacharya, Surajit Bhattacharjee, Saikat Majumder,
Sayantan Banerjee & Subrata Majumdar

Division of Molecular Medicine, Bose Institute, P-1/12, CIT Scheme VIIM, Kolkata 700 054, India

Received 24 June 2013; revised 26 August 2013

In experimental visceral leishmaniasis the causative obligate protozoan parasite, L. donovani invades and multiplies inside of macrophages, one of the sentries of the mammalian immune system. The initial host-parasite interaction between the Leishmania promastigote and the macrophage takes place at the plasma membrane interface. To trace any possible interaction between Toll-like receptor 2 (TLR2) and CC chemokine receptor 5 (CCR5) during early Leishmania-macrophage interactions, it was observed that the expression of both TLR2 and CCR5 were significantly increased, along with their recruitment to the lipid raft. TLR2 silencing attenuates CCR5 expression and restricts L. donovani infection, indicating a regulatory role of TLR2 and CCR5 during infection. Silencing of CCR5 and TLR2 markedly reduced the number of intracellular parasites in macrophages by host protective cytokine responses, while raft disruption using β-MCD affected TLR2/CCR5 cross-talk and resulted in a significant reduction in parasite invasion. In vivo RNA interference of TLR2 and CCR5 using shRNA plasmids rendered protection in Leishmania donovani-infected mice. Thus, this study for the first time demonstrates the importance of TLR2/CCR5 crosstalk as a significant determinant of Leishmania donovani entry in host macrophages.

 

 

Indian Journal of Experimental Biology

Vol. 52, January 2014, pp. 30-35

 

 

Response of male mice to odours of female mice in different stages of oestrous cycle: Self-grooming behaviour and the effect of castration

Shanmugam Achiraman1,2*, Devaraj SankarGanesh1, Soundarapandian Kannan3,
Soundararajan Kamalakkannan1, Natarajan Nirmala4 & Govindaraju Archunan2*

1 Department of Environmental Biotechnology,

2 Centre for Pheromone Technology, Department of Animal Science, Bharathidasan University, Tiruchirappalli 620 024, India

3 Department of Zoology, Periyar University, Salem 636 011, India

4 Department of Zoology, Periyar E.V.R College, Tiruchirappalli 620 023, India

Received 12 December 2012; revised 20 September 2013

The behavioural assays were carried out in a Y-maze wherein intact, castrated and testosterone-treated male mice were exposed to oestrus and non-oestrus urine samples. The intact male mice investigated more frequently and spent more time in the Y-maze arm with oestrus urine than in that with non-oestrus urine. In contrast, the castrated mice were not attracted to oestrus urine, whereas testosterone-treated mice showed preference for oestrus urine. The rate of self-grooming was higher in intact males in case of exposure to oestrus urine while the rate was lower with respect to non-oestrus urine. However, castrated mice exhibited less self-grooming behaviour which was partially restored by testosterone treatment. The results suggest that self-grooming behaviour is an indicator of detection and discrimination of oestrus by males, and supports the androgen role in male chemosensory ability to discriminate between oestrus and non-oestrus female odours.

 

 

Indian Journal of Experimental Biology

Vol. 52, January 2014, pp. 36-45

 

 

Hypolipidemic and antioxidant activity of ethanolic extract of
Symplocos racemosa Roxb. in hyperlipidemic rats: An evidence of
participation of oxidative stress in hyperlipidemia

 

A M Durkar1, R R Patil2, S R Naik*2

1Department of Pharmacology, Sinhgad College of Pharmacy, Pune, India

2Department of Pharmacology, Sinhgad Institute of Pharmlaceutical Science, Lonavala, Pune 410 401, India

Received 29 March 2013; revised 24 July 2013

Hypolipidemic and antioxidant activity profiles of ethanolic extracts of Symplocos racemosa (EESR) were studied by triton-WR1339 (acute) and high fat diet induced (chronic) hyperlipidemic rat models. In both the models, a significant increase in total cholesterol (TC), triglycerides (TG), very low density lipoproteins (VLDL), low density lipoproteins (LDL) and decrease in high density lipoproteins (HDL) in serum were observed. EESR (200 and 400 mg/kg) and simvastatin (10 mg/kg) administered orally reduced the elevated serum lipids (TC, TG, VLDL, LDL), restored the decreased HDL and improved the atherogenic index. In high fat diet induced hyperlipidemic model, EESR treatment prevented the increased formation of malondialdehyde (MDA) in liver, restored the depleted liver antioxidants, glutathione, superoxide dismutase, catalase significantly. The increased liver cholesterol, HMG-CoA reductase activity and body weight of hyperlipidemic rats were significantly reduced by EESR treatment. The EESR inhibited HMG-CoA reductase, a rate limiting enzyme in cholesterol biosynthesis, thereby causing hypolipidemic effects. EESR treatment also improved histoarchitecture of hepatocytes in hyperlipidemic rats. Experimental findings demonstrated anti-hyperlipidemic and antioxidant activity of EESR, which may be directly or indirectly related to its antioxidant activity. The hypolipidemic activity of EESR may be due to the presence of flavonoids phenolic compounds, phenolic glycosides and steroids.

 

 

Indian Journal of Experimental Biology

Vol. 52, January 2014, pp. 46-52

 

 

Antidiabetic activity of a triterpenoid saponin isolated from Momordica cymbalaria Fenzl

Raju Balwanth Koneri*, Suman Samaddar & Channakeshava Thimmasandra Ramaiah

Department of Pharmacology, Karnataka College of Pharmacy, Bangalore 560 064, India

Received 8 January 2013; revised 2 August 2013

Glucose uptake by isolated diaphragms of both diabetic, following streptozotocin administration, and non-diabetic animals increased in presence of an oleanane-type triterpenoid saponin isolated from the roots of M. cymbalaria. Insulin release was augmented by the presence of the saponin of M. cymbalaria (1 mg/mL) in rat insulinoma cell line (RIN-5F) pre-exposed to adrenaline (5 ΅M) and nifedipine (50 ΅M). Pancreatic histology also indicated considerable quantitative increase in β-cells (75%) when treated with the saponin. The results suggest that the saponin of M. cymbalaria possesses potential antidiabetic activity with respect to insulin secretion, which may be attributed to modulation of calcium channel, and β-cell rejuvenation.

 

 

Indian Journal of Experimental Biology

Vol. 52, January 2014, pp. 53-59

 

 

Behavioural and neuroendocrine effects of aqueous extract of Boerhaavia diffusa Linn. in mice using tail suspension and forced swim tests – A preliminary study

Dinesh Dhingra* & Rekha Valecha

Department of Pharmaceutical Sciences, Guru Jambheshwar University of Science and Technology, Hisar 125 001, India

Received 11 February 2013; revised 22 July 2013

The present study was done to evaluate the effect of aqueous extract of B. diffusa on depression in mice using behavioral models such as tail suspension test (TST) and forced swim test (FST). The extract (50, 100 and 200 mg/kg, po) was administered for 14 successive days to Swiss young albino mice. On 14th day, 60 min after administration, mice were subjected to TST and FST. The administration of aqueous extract of B. diffusa (50, 100 and 200 mg/kg, po) significantly decreased immobility period in both TST and FST, indicating significant antidepressant-like activity. The lowest dose (50 mg/kg) of the extract decreased the immobility period most significantly in FST, showing most potent antidepressant-like action. The efficacy of the extract (50 mg/kg) was comparable to fluoxetine (20 mg/kg). The extract did not show any significant effect on locomotor activity. The extract showed significant monoamine oxidase -A inhibitory activity. There was no significant effect of the extract on plasma corticosterone levels. Prazosin (α1-adrenoceptor antagonist), sulpiride (selective D2-receptor antagonist), baclofen (GABAB agonist), and p-CPA (tryptophan hydroxylase inhibitor) significantly attenuated the extract-induced antidepressant-like effect, when tested in TST. The extract might produce antidepressant-like effect by interaction with α1-adrenoceptors, dopamine-D2 receptors, serotonergic, and GABAB receptors. Thus, aqueous extract of B. diffusa showed significant antidepressant-like activity in mice probably through involvement of monoaminergic and GABAergic systems.

 

 

Indian Journal of Experimental Biology

Vol. 52, January 2014, pp. 60-66

 

 

Quercetin and β-sitosterol prevent high fat diet induced dyslipidemia and hepatotoxicity in Swiss albino mice

Kunal Sikder, Nilanjan Das, Swaraj Bandhu Kesh & Sanjit Dey*

Department of Physiology, University of Calcutta, 92, A. P. C Road, Kolkata 700 009, India

Received 14 March 2013; revised 23 July 2013

High fat diet group showed a significant rise in serum and hepatic total cholesterol, triglyceride and atherogenic index which are major biomarkers of dyslipidemia and cardiovascular risk. The liver function markers, lipid peroxidation and proinflammatory cytokine levels were elevated in high fat diet group whereas antioxidant levels significantly reduced. These findings manifest hepatic damage which was further confirmed by histological findings. Quercetin and β-sitosterol though structurally different yet both ameliorate the sickening changes in different mechanism. The current investigation is perhaps the first report of the mechanistic role of two polyphenols over dyslipidemia and subsequent hepatotoxicity.

 

 

Indian Journal of Experimental Biology

Vol. 52, January 2014, pp. 67-72

 

 

Human malaria in C57BL/6J mice: An in vivo model for chemotherapy studies

K Sunita*, M Rajyalakshmi , K Kalyan Kumar , M Sowjanya , PVV Satish & D Madhu Prasad

Department of Zoology & Aquaculture, Acharya Nagarjuna University, Nagarjunanagar 522 510, Guntur, India

Received 28 February 2013; revised 26 August 2013

The present work deals with the development of Plasmodium falciparum stages in mouse model and its potential for the study of efficacy of antimalarial drugs. C57BL/6J mice were infected with multidrug resistant P. falciparum strain then treated with arteether and artesunate. A response was observed to antimalarial drugs in terms of decrease in parasitemia. Mice infected with P. falciparum strain were successfully cured after treatment with either arteether or artesunate. The speed of parasite clearance time and burden of parasitemia differed for each drug and matched the previously reported observations, hence stressing the relevance of the model. These findings thus suggest that P. falciparum. infected human RBC (iRBC) – C57BL/6J mice can provide a valuable in vivo system and should be included in the short list of animals that can be used for the evaluation of P. falciparum responses to drugs.

 

 

Indian Journal of Experimental Biology

Vol. 52, January 2014, pp. 73-79

 

 

A common HPLC-PDA method for amino acid analysis in insects and plants

 

M K Dhillona*, Sandeep Kumarb & G T Gujar

aDivision of Entomology, Indian Agricultural Research Institute (IARI), New Delhi 110 012, India

bBiochemistry Laboratory, Germplasm Evaluation Division

National Bureau of Plant Genetic Resources, New Delhi 110 012, India.

Received 16 May 2013; revised 29 August 2013

A common method for analysis of 17 amino acids from various insect species and plant parts was standardized using HPLC-PDA. Prior to hydrolysis, lyophilization of test samples was found indispensible to remove excess moisture, which interferes in hydrolysis and separation of amino acids. After the hydrolysis of plant and insect samples, 500 and 100 ΅L of boiling HCl, respectively for reconstitution, and 20 ΅L of hydrolyzed samples used for derivatization, provided best results. Gradient profile of mobile phase and run time up to 65 min were standardized to (i) overcome the problems related to eluting underivatized sample part, (ii) optimize the use of mobile phase and run time, and (iii) get better separation of different amino acids. Analysis of Chilo partellus larvae reared on sorghum seedling powder based artificial diet indicated that arginine and histidine quantities were on par in both samples. However, methionine was higher, and leucine, isoleucine, lysine, phenylalanine, threonine and valine were lower in sorghum seedlings than in C. partellus larvae, suggesting compensation of these amino acids by the insect through voracious feeding, as is being expected from artificial diet. This method was found highly sensitive, reproducible and useful for the analysis of amino acids for better understanding of insect-plant interactions.

 

 

Indian Journal of Experimental Biology

Vol. 52, January 2014, pp. 80-88

 

 

Establishment of an in vitro plantlet regeneration protocol for unique varieties of brinjal (Solanum melongena L.) var. Mattu Gulla and Perampalli Gulla

 

A Muthusamy, KS Vidya, PK Pratibha, M Radhakrishna Rao, SB Vidhu, KP Guruprasad, U Raghavendra,
PM Gopinath & K Satyamoorthy*

Division of Biotechnology, School of Life Sciences, Manipal University, Planetarium Complex, Manipal 576 104, India

Received 17 July 2013; revised 20 August 2013

Brinjal (Solanum melongena L.) var. Mattu Gulla (MG) and var. Perampalli Gulla (PG) are unique varieties with distinct flavour cultivated in Udupi, Karnataka State, and are exposed to several biotic and abiotic stresses. An efficient and reproducible in vitro regeneration method is required to expedite the manipulation of these brinjal varieties to cope up with stress by tissue culture and gene transfer methods. The present study, reports a rapid and efficient in vitro regeneration protocol for these two varieties. The in vitro growth response was studied on Murashige and Skoog (MS) medium supplemented with 2, 4-D, BAP and IAA, and the plantlets were regenerated efficiently from callus cultures of leaf, cotyledon and hypocotyl explants. Among the three explants, the hypocotyl explants were found to have better callus induction and multiple shoot regeneration. High frequency of shoot initiation was achieved from hypocotyl derived calluses in MS media with 2.0 mg/L BAP and 0.5 mg/L IAA in MG and PG. Efficient and rapid shoot proliferation, and elongation were noted in MS medium with 1.0 mg/L BAP and 0.3 mg/L GA3. The in vitro regenerated shoots produced healthy roots when they were cultured on MS medium supplemented with 0.5 mg/L IBA. A significant difference was observed in percentage of callus induction, number of shoots per callus, shoot elongation and number of hardened plantlets of MG and PG. MG showed maximum response in all stages of culture than PG. Hardening of plantlets in tissue culture was achieved in three weeks. The hardened plantlets were grown in pots for further acclimatization in green house and finally transplanted to experimental garden where they developed into flowering plants and produced mature fruits with viable seeds.

 

 

Indian Journal of Experimental Biology

Vol. 52, January 2014, pp. 89-96

 

 

Potential application of β-1, 3 glucanase from an environmental isolate of Pseudomonas aeruginosa MCCB 123 in fungal DNA extraction

 

Divya Jose, P Jayesh, Prem Gopinath, A Mohandas & I S Bright Singh*

National Centre for Aquatic Animal Health, Cochin University of Science and Technology, Lakeside Campus,
Fine Arts Avenue, Cochin 682 016, India

Received 7 March 2013; revised 8 August 2013

Pseudomonas aeruginosa MCCB 123 was grown in a synthetic medium for β-1,3 glucanase production. From the
culture filtrate, β-1,3 glucanase was purified with a molecular mass of 45 kDa. The enzyme was a metallozyme as its β-1,3 glucanase activity got inhibited by the metal chelator EDTA. Optimum pH and temperature for β-1,3 glucanase activity on laminarin was found to be 7 and 50 °C respectively. The MCCB 123 β-1,3 glucanase was found to have good lytic action on a wide range of fungal isolates, and hence its application in fungal DNA extraction was evaluated. β-1,3 glucanase purified from the culture supernatant of P. aeruginosa MCCB 123 could be used for the extraction of fungal DNA without the addition of any other reagents generally used. Optimum pH and temperature of enzyme for fungal DNA extraction was found to be 7 and 65 °C respectively. This is the first report on β-1,3 glucanase employed in fungal DNA extraction.