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Tissue Research » Past Abstracts
1.  Journal of  Cell and Tissue Research 13(1): 3431-3438  (2013)

Effect of  antioxidants (alpha-lipoic acid and bamboo shoot extract, either alone  or in combination), in lead induced oxidative stressed animals

Kalia, K., Chiragini, H.M. and  Sood, P.P.

BRD School of  Biosciences, Sardar Patel University, Vallabh Vidyanagar 388 120
E. Mail: kirankalia_in@yahoo.com

Abstract: The present study has been carried out to investigate the protective role of the bamboo shoot methanolic extract (BLME) against lead induced hepatic oxidative insult. Lead acetate at a dose of 12.5  mg/ kg body weight (b.w.) per day was preferred as the source of lead  in our study. We have included various liver cell injury markers like serum glutamate pyruvate transaminase (SGPT), serum glutamate oxaloacetate transaminase (SGOT) and oxidative stress markers as a parameter to measure degree of oxidative damage viz. advanced oxidation end product (AOPP), reduced glutathione (GSH), thiobarbituric acid reactive substances (TBARS), and healing due to bamboo shoot  extract. Activities of enzymes, like superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GPx), responsible for antioxidant defense system were also estimated. Bamboo shoot  extract at three different doses i.e. 50 mg, 250 mg and 500 mg per kg b.w. for last fifteen days after arsenic treatment prior to sacrifice altered most of the above parameters of oxidative stress caused by lead. Bamboo shoot  extract eliminate the effect of lead induced oxidative stress by being a potent free radical scavenger.

Key words: Bamboo shoot,  Lead, Hepatic protection


2.  Journal of  Cell and Tissue Research 13(1): 3439-3443  (2013)

JAK2 V617F mutation: a marker for CML progression?

Manjula, G., Prajitha, E.M., Sailaja, K., Raghunadha  Rao, D., Anuradha, C., Sugunakar, V.,   Sandhya, A.,  Nageswara Rao, D. and Vishnupriya, S.

Department of  Genetics, Osmania University, Hyderabad  500 007.  E. mail: sattivishnupriya@gmail.com

Abstract: Treatment with Imatinib mesylate leads to molecular remission in majority of Philadelphia chromosome (Ph) positive Chronic Myeloid Leukemia (CML) cases. However, sometimes persistent anemia or evolving myelofibrosis is observed due to a co-existing Ph negative chronic Myelo Proliferative Disorder (CMPD) clone. Polycythemia vera (PV) is the most common Myelo Proliferative Disorder (MPD) coexisting with CML and 80% of cases with PV showed JAK2V617F mutation. Janus kinase2 (JAK2) gene plays a major role in maintaining BCR-ABL stability as BCR-ABL exerts its oncogenic signaling through the involvement of Jak2 phosphorylation. JAK2V617F mutation results in hyperactivation of JAK2 gene and had been implicated in several hematologic malignancies. In our study, the JAK2V617F mutation was detected through Amplification Refractory Mutation Detection-Polymerase Chain Reaction (ARMS-PCR) method in 6 out of 480 CML cases. None of the 350 controls showed this mutation. Patients with JAK2 mutation except for one failed to show complete molecular remission despite the prolonged treatment with Imatinib. The results suggested that JAK2 V617F mutation could be a marker for CML progression and JAK2 could serve as alternative drug target in Imatinib resistant CML cases with persistent myelofibrosis.

Key words: CML, BCR-ABL, Imatinib mesylate, JAK2 V617F, ARMS- PCR


3.  Journal of  Cell and Tissue Research 13(1): 3445-3449  (2013)

Electron microscopic radioautopgraphic study on mitochondrial protein synthesis in colonic epithelial cells of aging mice

Nagata, T.

Department of Anatomy and Cell Biology, Shinshu University School of Medicine, Matsumoto, 390- 8621
Japan and Department of Anatomy, Shinshu Institute of Alternative Medicine, Nagano,  380-0816,  Japan. E. mail: nagatas@po.cnet.ne.jp

Abstract: For the purpose of studying the aging changes of macromolecular synthesis in the colonic cells of experimental animals, we studied 10 groups of aging mice during aging from fetal day 19 to postnatal month 24. They were injected with 3H-leucine, a precursor for protein synthesis, sacrificed and the colonic tissues were processed for light and electron microscopic radioautography. On many radioautograms the localization of silver grains demonstrating protein synthesis in colonic epithelial cells in respective aging groups were analyzed qualitatively. The results revealed that the number of mitochondria increased from embryonic day 19 to postnatal adult month 1 and 2, reaching the maximum, then decreased to senile year 1 to 2. On the other hand, the number of labeled mitochondria showing protein synthesis increased from embryonic day 19 to postnatal newborn days, adult month 1 and 2, reaching the maximum and decreased to month 24. To the contrary, the labeling index increased from embryonic day 19 to postnatal day 3, reaching the maximum, then decreased to day 14, increased to month 1, decreased to month 6, increased to month 12 and decreased to month 24, indicating the aging changes. These results demonstrated that intramitochondrial protein synthesis in the colonic epithelial cells increased and decreased again and again for 3 times repeatedly due to aging of individual animals depending upon the cellular activities at respective aging stages

Key words: Mitochondria, Colonic epithelia, EM radioautography, Protein synthesis


4.  Journal of  Cell and Tissue Research 13(1): 3451-3454 (2013)

In-vivo study of tissue distribution of tamoxifen  loaded stealth liposomes in mammary tumor

Mali, D.S., Nanjwade, B.K., Unger, B.S. and  Manvi, F.V.

Department of  Pharmaceutics, KLE University’s College of Pharmacy, Belgaum-590 010, Karnataka
E. mail: deepakmali.pharm@gmail.com Cell: 094481 34368

Abstract: This study was carried out to evaluate and compare the tissue distribution profile of tamoxifen citrate, when administered intravenously (i.v.) as a simple solution or when encapsulated into stealth liposomes with an average diameter of 479 nm. Passive targeting by stealth liposomes, once combined with efficient intracellular delivery, may be a very useful strategy to improve the antitumor efficacy of the anticancer agents. Natural ability of liposomes to target cancer cells (tumor) due to enhanced permeability and retention (EPR) effect and polyethylene glycol (PEG) coating of stealth liposomes which is inert in the body, allows for longer circulatory life for the drug delivery mechanism results in enhanced accumulation of tamoxifen in tumor. Tamoxifen loaded stealth liposomes (TSL) were prepared with thin film hydration method by using cholesterol, DMPC, DSPC, and polyethylene glycol-4000 (PEG-4000) in order to achieve prolonged circulation time and sustained release.  In vivo tissue distribution study was carried out in Sprague-Dawley rats having mammary tumor induced by chemical carcinogen DMBA (7,12-Dimethylbenz(a)anthracene). After intravenous administration, the primary site of accumulation for the TSL was tumor accounting up to 36% after 6 h post injection compared to 6% observed with non-formulated tamoxifen. The results indicate that, TSL may present a promising delivery system to achieve preferential tumor-targeting.

Key words: Tamoxifen, Polyethylene glycol, Stealth, Liposome


5.  Journal of  Cell and Tissue Research 13(1): 3455-3458 (2013)

Molecular characterization of alpha toxin gene of Clostridium perfringens from chicken meat

Nithin Prabhu, K., Wilfred Ruban, S., Naveen, B. R. and Raghunath, B. V.

Department of  Veterinary Microbiology, Veterinary College, KVAFSU, Hebbal, Bangalore  560 024,
Karnataka.  Email: nithinprabhuk@yahoo.com Mobile: +91 9844995078

Abstract: Present research contribution aims to investigate the prevalence of Clostridium perfringens in chicken meat obtained from retail outlets in and around Bangalore, Karnataka. In total 71 Chicken meat samples were collected and were processed for the isolation and identification of C. perfringens. Bacteriological investigation revealed the presence of C. perfringens in 58 samples (81.69 %) based on biochemical characterization. All these isolates were subjected to virulence determination for the presence of alpha (α) toxin gene by Polymerase chain reaction [PCR]. This PCR determination of virulence genes suggested that chicken meat sample may be considered a significant source C. perfringens. The high incidence of this bacterium in poultry meat may indicate insanitary conditions and improper handling at processing area.

Key words: Chicken meat, Clostridium perfringens, alpha toxin.


6.  Journal of  Cell and Tissue Research 13(1): 3459-3464 (2013)

Asiaticoside counters the parkinsonic variations in mice

Uvarajan, S.  and  Vanisree, A.J.

Department of Biochemistry, Indo-American College, Cheyyar, Tamilnadu; Tamilnadu. E. mail: uvasambath@gmail.com

Abstract: To evaluate the neuroprotective effect of asiaticoside a triterpenoid isolated from Centella asiatica against MPTP (1 – methyl 4 – phenyl 1,2,3,6 tetrahydropyridine) induced neurotoxicity in experimental mice. Mice were divided in to 4 groups : Group I received vehicle saline, Group II was treated with 20mg/kg of body weight of MPTP (2 doses with 2 h intervals), Group III received MPTP along with 50 mg/kg body weight of asiaticoside for  21 consecutive days starting from the day of MPTP intoxication. Group IV received 50 mg/kg body weight of asiaticoside for the same period serving as drug control. Animals were sacrificed at the end of experimental period. The striatum was taken and analyzed for enzymatic and non-enzymatic antioxidants and histopathological changes which could provide clues to the extent of neurodegeneration. Study shows that superoxide dismutase, catalyse, lipid peroxidation, reduced glutathione, glutathione peroxidase, vitamin C and E, total lipid, cholesterol  and Triglycerides were altered in MPTP treated mice.  The changes were found to be reversed significantly (p<0.05) on treatment with asiaticoside. The biochemical studies were supported by the histological observations in the striatal tissues. The observations suggest that asiaticoside may prove to be efficacious in protecting neurons from the oxidative damage caused by the insult of MPTP.

Key words: Centella asiatica, Asiaticoside, Parkinson disease


7.  Journal of  Cell and Tissue Research 13(1): 3465-3471 (2013)

Dynamics of dichotomous spermiogenesis in Spodoptera mauritia boisd (Lepidoptera: Noctuidae)

Manogem, E.M. and  Nair, V.S.K.

Department of  Zoology, University of Calicut, Thenhipalam, Malappuram- Kerala.
E. mail: manogemvinod@yahoo.com Phone: +91-9895325917

Abstract: The ultrastructural changes in the germ cells of S. mauritia testis, occurring during the last larval (sixth instar) and pupal stages is reported here. Germ cells of both larval and pupal stages comprises of spermatidal cysts.  During spermatogenesis, the spermatids are differentiated into typical eupyrene (nucleated) and atypical apyrene (anucleated) sperm bundles. Spermatid cysts of pharate pupa noticed elongated nucleus, with adjacently placed acrosomal bag and formation of single nebenkern, by fusion of mitochondria.  Early eupyrene spermatid cyst of day 1 pupa comprised mainly of nucleus, an axial filament/axoneme, multivesicular bodies with Golgi complex.  The single structure called nebenkern, later on divide to form unequal mitochondrial derivatives. The  axoneme possess, a typical 9+9+2 microtubular arrangement.  Likewise, apyrene spermatid cysts are characterized by the formation of nebenkern.  During later stages of apyrene spermiogenesis, acrosome formation is not detected and basal body is not attached to the nucleus.  Eupyrene spermatozoa are distinguished from apyrene ones by the possession of an elongated nucleus, with two apposed mitochondrial derivatives fusing into a crescent shaped structure, with lacinate and reticulate appendages, and an axoneme with 9+9+2 configuration.  Apyrene sperm is small, when compared to eupyrene.

Key words: Spodoptera mauritia, Spermatogenesis


8.  Journal of  Cell and Tissue Research 13(1): 3473-3478 (2013)

SEM study of adhesive organ of typical hill stream fish Garra gotyla (Teleostei; cyprinidae)

Gaur, K. S., Sharma, V.  Verma, B. K. and  Sharma, M. S.

Limnology & Fisheries Research Laboratory, Department of  Zoology, University College of Science,
M. L. Sukhadia University, Udaipur 313 001, Rajasthan. E. mail: madhu_limnology@yahoo.co.in

Abstract: The study of adhesive organ (AO) of the typical hill stream fish Garra gotyla inhabiting the hill streams of South-eastern Rajasthan was carried out for surface image and morphological details. The scanning electron microscopy (SEM) revealed that the fish adhesive organ (AO) has upper and lower fringed lips bearing tubercles beset with two types of spines which are modified squamous epithelium cells. The adhesive organ (AO) secretes mucus which displaces water and air to create vacuum helping fish to fasten with substratum. From the study it appears that spine and mucus collectively function and realize adhesion in the speedier waters of the streams.

Key words: Garra gotyla, Adhesive organ


9. Journal of  Cell and Tissue Research 13(1): 3479-3484 (2013)

Venom in a sting and In Silico prediction of antigenic determinants of venom of Apis dorsata

Hemke, V.  

Shri Shivaji College Akot-444101, Distric Akola, Maharashtra. E. mail:  vjshri@ymail.com

Abstract: The honeybee differs from other flying Hymenoptera from both entomologic and allergic viewpoints. Beekeepers are particularly at risk for honeybee exposure and allergy.  Bee venom is a complex mixture of proteins (enzymes and peptides) with unique pharmacological activities. The main enzyme in A. dorsata venom is phopholiphase A2. Honeybee venom is complex, and the delivery mechanism provides for a large but often variable amount of injected venom.  However, in present study is designed  to find out quantity of honeybee venom injected in a single sting by A. dorsata and in silico prediction of antigenic determinants of their proteins.

Key words: Honeybee venom, Antigenic determinants, In silico


10.  Journal of  Cell and Tissue Research 13(1): 3485-3490 (2013)

Purification and identification of Aspergillus niger induced novel protein from groundnut (Arachis hypogeal L.)

Jajda, H.M., Patel, R.R. and Thakkar, V.R.

Food Quality Testing Laboratory, Navsari Agricultural University, Navsari 396 450;
E. mail: harsurbiochem@gmail.com  Phone 08128699171

Abstract: A 110-kDa protein was over-expressed in groundnut (Arachis hypogaea L.) seedlings due to infection of Aspergillus niger. Densitometric quantification revealed that the protein start inducing after 4hr post infection and maximum induction is seen after 42hr. When subjected to SDS page, protein showed three subunits.  The purification  of induced protein from maximum induced fraction was carried out using ammonium sulphate precipitation followed by size exclusion chromatography using shephadex G- 100. Purified tri-peptide were confirmed by  two-dimensional gel and  subjected to tryptic digestion, followed by identification using matrix-assisted laser desorption/ionization–time of flight mass spectrometry. This sequence segment was searched for homologous sequences using MASCOT data base program and no similarity was found with other reported sequences of groundnut proteins. Even when the expected value parameters were increased, it yielded the same result. The searches were done against non redundant database. Further, no match to this sequence was found from a search on the Express Sequence Tag. This shows that the sequence may be unique. Thus, we assume that induced protein is a unique and novel protein which is not reported earlier in groundnut plants.

Key words: Groundnut, Aspergillus niger, Unique Protein


11. Journal of  Cell and Tissue Research 13(1): 3491-3493 (2013)

Electrophoretic patterns of esterases of parotoid gland of common indian toad Bufo melanostictus (Schneider)

Raju, N. and  Venkaiah, Y.

Department of Zoology, Kakatiya University, Warangal 506 009. E. mail: venkaiah_yanamala@
rediffmail.com, Mobile No. +91-9849848705

Abstract: The present study was carried out to investigate the patterns of esterase isoenzymes (commonly known as carboxyl esterases) in parotoid gland secretion in Bufo melanostictus and its extraction through electrophoresis. The results revealed that the activity of individual esterase was not completely inhibited by pCMB and physostigmine. However, a complete inhibition was observed in the presence of Paraxon an Organo phosphate.

Key words: Bufo melanostictus, Esterases.


12. Journal of  Cell and Tissue Research 13(1): 3495-3498 (2013)

Free radicals  spare  KCL  but reversibly impair ne mediated  vascular contractility in goat isolated pulmonary artery in vitro

Sharma, D.K., Telang, R.S. and  Varshneya, C.  

Department of Pharmacology and  Toxicology, COVAS, CSKHPKVV, Palampur (H.P.).  
 E. mail: dineshcovas79@gmail.com. Mobile : 09418477233

Abstract: The present study was designed to determine in vitro effect of free radicals on the pharmacological behavior of goat isolated pulmonary artery (GPA).  Isolated organ bath studies were undertaken on GPA stripes to determine the effect of hydroxyl (Fenton mixture) radicals on pre- and post-agonist (NE or KCl) exposure. Control experiments were done with individual ingredients of free radical generation mixture. Recovery studies were done to assess the nature of impairment. The free radicals had no effect on non-stimulated tissues. Pretreatment of  the tissues with free radicals abolished the NE induced contractions; while the KCl induced contractions remained unaffected. Similarly the free radicals relaxed the plateau phase of NE induced contractions, while again sparing the KCl induced plateau. This indicates a deleterious effect of free radicals on receptor proteins and second messenger systems. The recovery studies revealed that this impairment was transient and the tissue regained its contractile ability after sufficient rest period. The study reports a significant detrimental but reversible effect of free radicals on norepinephrine induced contractions, while sparing high K+ induced contractions in vitro.

Key words: Vascular contractility, Hydroxyl radicals, Pulmonary artery


13. Journal of  Cell and Tissue Research 13(1): 3499-3502 (2013)

Antihyperglycaemic and antioxidant effects of methanolic extract of Tinospora cordifolia in alloxan induced diabetic rats

Sharma, D.K., Varshneya, C. and Chauhan, S.

Department of  Pharmacology and Toxicology, COVAS, CSKHPKVV, Palampur (H.P.).
E. mail: dineshcovas79@gmail.com. Cell: 09418477233

Abstract: In the present study, methanolic extracts of Tinospora cordifolia (T. cordifolia) stem were administered orally @ 100 and 200 mg/kg body weight to alloxan induced diabetic rats for 15 days. This resulted in significant reduction (p<0.01) in blood glucose level in diabetic animals. The levels of blood glycosylated hemoglobin (HbA1c), serum AST and ALT significantly (p<0.01) increased in diabetic rats, however these levels returned to normal in T. cordifolia extract treated animals. There was a significant (p<0.01) increase in LPO and reduction in the activities of antioxidant enzymes CAT, GSH & SOD   in diabetic rats. The administration of methanolic extracts of T. cordifolia (200) significantly reversed the condition. CAT level also elevated significantly in treated animals. However, glibenclamide restored blood glucose, HbA1c, ALT, AST, LPO, CAT and SOD and the restoration was greater than the treated groups. The study, therefore, indicated that T. cordifolia possesses antidiabetic activities along with good antioxidant properties and can be used as an adjunct therapy in diabetes.

Key words:   Diabetes, Alloxan, Tinospora cordifolia


14. Journal of  Cell and Tissue Research 13(1): 3503-3506 (2013)

Economic, easy and fast method for isolating high quantity and quality DNA from cotton (Gossypium spp) suitable for molecular studies

Sangannavar, P.A., Vanti, G.L., Anupama, S., Methre, R.M., Bhute, N.B., Savita, S.G., Vamadevaiah, H. M. and Katageri, I.S.

Agricultural Research Station, University of Agricultural Sciences, Dharwad -580007, Karnataka.
E. mail: dr.sangannavar@gmail.com

Abstract: The extraction of DNA from cotton is difficult due to the presence of secondary compounds. In this study an effort was made to have easy, rapid and economical protocol for isolation of good quality and quantity DNA.  This method involves no expensive equipment and chemicals, require fewer steps, tubes and allows DNA extraction from many samples per day per person, making this method more economical. DNA yield from this protocol was incredibly high ranging up to 1000 μg/g of fresh sample. Purity of extracted DNA was excellent as A260/A280 ratio was 1.88-1.97 and A260/A230 ratio was > 2, suggesting that the preparations were sufficiently free of proteins and polyphenolic compounds, respectively. Cotton DNA extracted using this method is completely digested with restriction enzymes and supports PCR.

Key words: DNA extraction, Gossypium, Cotton.


15.  Journal of  Cell and Tissue Research 13(1): 3507-3511 (2013)

Evaluation of antimicrobial efficacy of aqueous extract and essential oil of clove (Eugenia caryophylata Thunb) in chicken meat against  various food born pathogens

Shekar R. Badhe., Fairoze, M. N.,  Raghunath, B.V., Sudharshan. S.,  Wilfred Ruban , S. and Chandrashekaran, V.

Department of  Livestock Products Technology, College of Veterinary Science and Animal Husbandry, JAU, Junagadh  362 001.  E. mail: drshekhar15@gmail.com

Abstract: A study on antimicrobial efficiency of aqueous extract and essential oil of clove on food borne pathogens was carried out.. 100 per cent clove aqueous extract was found to be more effective against Staphylococcus aureus and Bacillus cereus. Bacillus cereus and E coli, which were most sensitive to the action of essential oil of clove, Salmonella typhimurium, E. coli O 157 and Klebsiella pneumoniae, which were moderately sensitive, whereas, Bacillus subtilis and field isolates of Salmonella, E coli and Staphylococcus aureus were comparatively less sensitive to the action of essential oil of clove. Dilutions of essential oil of clove at 1:150 and 1: 250 brought about a significant decrease in bacterial counts compared to 1:500.

Key words: Clove, Essential oil, Food borne pathogens


16. Journal of  Cell and Tissue Research 13(1): 3513-3517  (2013)

Regeneration protocol for chilli  (Capsicum  annuum L.) variety Mathania

Rizwan, M., Sharma, R., Soni, P., Gupta, N.K.  and Singh, G.

Plant Biotechnology Centre, Swami Keshwanand Rajasthan Agricultural University, Bikaner
 Rajasthan. E. mail: nkgupta69@yahoo.co.in  Tel.: 09460987039

 Abstract: A protocol for regeneration has been developed for chilli (Capsicum annum L. variety Mathania). High frequency regeneration of shoot buds from cotyledonary leaves and hypocotyls was achieved on Murashige and Skoog  (MS) medium supplemented with 10 mgL-1 BAP and 1 mgL-1 IAA. Cytokinine free MS media  was found competent for further growth and elongation of regenerated shoots. Both elongation of shoot buds and subsequent rooting was obtained on MS medium supplemented with 1 mgL-1 IAA. Rooted shoots were transplanted in magenta box filled with soilrite, where they grew well and attained maturity. Regenerated plants were phenotypically normal. The use of this protocol opens prospects for transformation in Mathania variety of chilli in to save it from leaf curl virus infestation.

Key words: Capsicum annum L., Tissue culture regeneration.


17. Journal of  Cell and Tissue Research 13(1): 3519-3524  (2013)

Effect of dried fruit extract of Benincasa hispida on brain behaviour in laboratory animals

Ambikar, D.B. and  Mohanta, G. P.

Karpagam University, Coimbatore-641021, Tamilnadu and Marathawada Mitra Mandal’s College of
 Pharmacy, Thergaon, Pune 411033 .  E. mail: pharmascholy@gmail.com

Abstract: The present investigation was aimed at determining the spectrum of neuropharmacological activity of different doses (100, 200, 400 mg/kg) of petroleum ether (BHP), methanolic (BHM) and aqueous extract (BHA) obtained from Benincasa hispida in laboratory animals. Benincasa hispida was studied for its effect on motor coordination, locomotor activity, cognitive behavior, anxiolytic activity, haloperidol induced catalepsy and anticonvulsant activity. The BHM and BHA 400 mg/kg showed significant (P< 0.01) decrease in locomotor activity and exploratory behavior but no effect on motor-coordination. Same doses showed significant (P< 0.01) increases in discrimination index in object recognition test. BHM 400 found to prolong haloperidol induced catalepsy in mice (P< 0.05). Moreover the BHM and BHA 400 mg/kg extracts also showed significant (P< 0.05) analgesic activity in hot plate method. The result points towards the potential activity of the Benincasa hispida as anxiolytic, analgesic and nootropic activity.

 Key words: Benincasa hispida, Anxiolytic, Anticonvulsant


18. Journal of  Cell and Tissue Research 13(1): 3525-3530  (2013)

In vitro and in vivo antioxidant activities of Seabuckthorn berries and seed oil

Chauhan, S., Varshneya, C. and Ghabru, A.

Department of  Pharmacology and Toxicology, College of Veterinary and Animal Sciences, CSK Himachal Pradesh Krishi Vishvavidyalaya, Palampur 176 062 (H.P). E. mail: shivanic9@gmail.com

Abstract: The present study was aimed at evaluating the Seabuckthorn (SBT) seed oil and fruit pulp for antioxidant effect in chromate induced oxidative stressed rats. The antioxidant activity of SBT seed oil and different pulp extracts (100% methanolic, 70% ethanolic, 50% methanolic and 100% aqueous) was assessed out by in vitro  models such as scavenging of free radicals using 1,1-Diphenyl-2-picryl-hydrazyl (DPPH), nitric oxide, 2,2-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid (ABTS), hydroxyl and superoxide. For in vivo studies oxidative stress was induced in male Wistar rats by oral administration of potassium-di-chromate (K2Cr2O7) 30mg/kg b.w. for 56 days. Effect of concurrent administration of SBT seed oil and lyophilized SBT pulp extract {(70% ethanolic extract (ETE)}, each at two doses of 500, 1000mg/kg b.w. and 100, 200mg/kg b.w. respectively, was also given by oral route. Lipid peroxidation (LPO), reduced glutathione (GSH), superoxide dismutase and catalase activity were determined in both liver and kidneys on day 56. Both SBT seed oil and pulp ETE produced free radical scavenging activity. There was a significant (P<0.05) decrease in lipid peroxidation in liver and kidneys of pulp ETE treated groups as compared to chromium treated groups. The SBT pulp ETE reverted the GSH level, SOD and catalase activity in both liver and kidney more towards the control as compared to oil treatment. On the basis of these observations it can be concluded that SBT pulp ETE has good in vitro as well as in vivo antioxidant activity as compared to seed oil.
 
Key words: Seabuckthorn, Pulp, Seed oil, Antioxidant


19. Journal of  Cell and Tissue Research 13(1): 3531-3534 (2013)

Gas chromatography-mass spectrometry analysis of methanol extract of Michelia champaca L. flowers

Ananthi, T. and  Chitra, M.

P. G. and Research Department of Biochemistry, S.T.E.T Women’s College, Mannargudi 614 001.
 E. mail: mschitra21@yahoo.com

Abstract: Michelia champaca (Magnoliaceae), commonly known as Svarna champa is a glorious ancient Indian medicinal plant was investigated for GC-MS (Gas Chromatography-Mass Spectrometry) analysis to determine the chemical constituents present in methanol extract of flowers. Powdered flower plant materials were subjected to successive extraction with methanol by Soxhlet extraction method. Totally, 27 different compounds from methanol extract were identified. All the compounds were medicinally valuable for the treatment of various human ailments. In addition, all the phytochemical compounds were needed further investigations on toxicological aspects for the development of new lead of therapeutic interest.

Key words: Michelia champaca flower, Phyto-chemical analysis


20. Journal of  Cell and Tissue Research 13(1): 3535-3540 (2013)

Cost effective plant regeneration protocol for rough lemon (Citrus jambhiri lush) using gum konda-gogu as gelling  agent in plant tissue culture media

Singh, B., Singh, J. and  Kaur, A.

Plant Tissue Culture Laboratory, Department of Biotechnology, Khalsa College Amritsar 143 002 (Punjab).  E. mail: bbs171@rediffmail.com
 
Abstract.  The goal of this study was to develop cost effective regeneration protocol for rough lemon using natural exudate gum from Cochlospermum gossypium (gum kondagogu) as gelling agent in plant tissue culture media. Nodal segments of rough lemon (Citrus jambhiri Lush) were cultured on MS medium supplemented with kinetin (KN) 2 mg/l and gelled with agar or gum kondagoggu alone and in combinations. Maximum 66.66% cultures showed shoot regeneration on 0.8% agar gelled medium (control) with an average shoot length of 0.74 cm. The shoot regeneration response on media gelled with 3% gum kondagogu was 63.88 % with an average shoot length of 0.79 cm.  Use of agar (0.1 to 0.3%) along with gum kondagogu (2.5 and 3%) improves shoot regeneration response but there is decrease in average shoot length. Regenerated shoots were rooted on MS medium supplemented with Indole-3-butyric acid (IBA) 2 mg/l and gelled with agar or gum kondagogu alone and in combinations. In agar gelled medium (control), 52.77 % cultures showed root regeneration with an average root length of 5.48 cm. In 3% gum kondagogu gelled medium maximum of 52.08 % cultures showed root regeneration with an average length of 5.53 cm. Addition of agar in 3% gum gelled medium decreases root regeneration response and average root length. Firmness of gum gelled medium was less as compared to agar gelled medium. However, there was no significant difference in regeneration response of nodal explants on agar (0.8%) and gum (3%) gelled medium. Cost of gum kondagogu is 6.5 times less than that of agar so it can be used as gelling agent to reduce production cost of tissue culture raised plantlets.

Key words: Citrus jambhiri, Gum kondagogu, Tissue culture



21. Journal of  Cell and Tissue Research 13(1): 3541-3544 (2013)

Non-enzymatic antioxidant expression and nutritional composition of Calocybe indica under different organic supplementations

Sharma, S. K. and Lall, A. M.

School of Biotechnology SKUAST, Jammu, Main Campus Chatha, Jammu 180 009 (J&K);
 E. mail: princesatish1@gmail.com  Cell:  09419281730

Abstract: The modulation of the mushroom used now-a-days in the field of medicine, which contains vitamins, nutrients, minerals etc. Among these, the “antioxidants” which are capable of deactivating the free radicals follow either enzymatic or non-enzymatic pathways. The aim of this work was to improve the nutrient composition   as well as antioxidant status (non enzymatic) of Calocybe indica (milky mushroom). The present study revealed that supplementation improves the production, non-enzymatic antioxidant and nutrients in the samples as compare to control. The striking feature of present work is to maximize the usage of mushroom as food to maintain health and got benefits against various dreadful diseases when there are certain unstable conditions for the usage of mushroom in the fresh form.

Key words: Calocybe indica, Non-enzymatic antioxidant


22. Journal of  Cell and Tissue Research 13(1): 3545-3548 (2013)

Improved protocol for in vitro propagation of gloxinia ( Sinningia sp.)

Sharma, S. K.  and  Sharma, M.

School of Biotechnology, SKUAST Jammu Main Campus Chatha 180 009 (J&K).
E. mail:  princesatish1@gmail.com   Cell:  09419281730

Abstract: An efficient and reproducible protocol for in vitro multiplication of gloxinia has been developed. Leaf discs used as explants were surface sterilized with HgCl2 (0.1%) and Bavistin (2%). Sterilized explants were cultured on MS media augmented with BAP (2 mg/l) and NAA (0.5 mg/l) which proved most appropriate for shoot induction. Sprouted shoots were sub cultured on medium with different concentrations of BAP which resulted in varying degrees of multiple shoots.  Increasing  the concentration of BAP resulted in reduction in number of shoots per explant. Maximum proliferation of shoots was observed in MS medium augmented with BAP (2.0 mg/l)  and NAA (0.5 mg/l)  within 2 weeks and  average number of shoots per explant was 7.3. The in vitro raised shoots  of gloxinia were successfully rooted in MS media fortified with NAA and IBA. Rooted plantlets could be successfully established in potting mixture of cocopeat and sand (1:1).

Key words: In vitro, Gloxinia propagation, Sinningia


23. Journal of  Cell and Tissue Research 13(1): 3549-3556  (2013)

Bacopa monniera herb as an antioxidant in reducing lipid peroxidation levels in cultured human endothelial cells

Almelkar, S.I., Divate, S. and  Patwardhan, A. M.

Tissue Culture Lab (Applied Medical Biology), First Floor Immunopathology CVTC, Department of  Cardiovascular and Thoracic Surgery, CVTS Building, Seth G. S. Medical College and  King Edward Memorial Hospital, Parel, Mumbai - 12, Maharashtra.
E. mail: shahdabalmelkar@gmail.com  

Abstract: Bacopa monnieri Linn. (Family Scrophulariaceae) is commonly known as Brahmi is a component of several popular drugs of ayurvedic system medicine and is clinically used for memory enhancing, insomnia and as mild sedative. Bacopa monniera extract (BME) is a natural antioxidant and used to cure many gerontological disorders but their effect on vascular cells and especially in cultured endothelial cells is unknown. Oxidative stress induced lipid peroxidation enhances the levels of malondialdehyde (MDA) in vascular cells which leads to blood vessels diseases such as atherosclerosis or stenosis. The main goal of this research was to investigate effect of BME on MDA levels in normal cultured human endothelial cells (HUVECs) and stress induced cultured HUVECs. We investigated antioxidant effect of BME by examining cell viability (MTT) assay. Oxidant effect H2O2 by examining cell viability (MTT) assay, immunocytochemistry for vonWillebrand Factor (vWF) expression with BME , H2O2 , Pre BME + Post H2O2 , Pre H2O2 +Post BME exposure, lipofuscin granule formation in H2O2 exposure and absence in BME exposure, lactate dehdyrogenase (LDH) release and comparative lipid peroxidation judged by comparing the levels of MDA. MTT assays provided us the selection range for BME (6.25μg-100μg) and oxidative stress ((H2O2 -6.25µM-100µM) concentration. Expression of vWF was up regulated in BME exposure while vWF expression was extremely down regulated in H2O2 exposed HUVECs. Pre BME + Post H2O2 exposure resulted in moderate expression of vWF while Pre H2O2 + Post BME showed improved expression of vWF. When these cells were exposed to H2O2 they  showed the presence of copious lipofuscin granule formation while BME exposure showed lack of lipofuscin granule. LDH release was slow down after exposure of BME (6.25µg -100µg) and protected cell against LDH release. We also found reduction of  MDA levels in stressed HUVECs during  dose-dependent exposures of BME.  The investigation shows that BME is an excellent natural antioxidant which plays a defensive role against free radical attack and reduce the lipid peroxidation  in stressed cultured HUVECs.

Key words: Bacopa monniera, Lipid peroxidation, Human endothelial cells


24. Journal of  Cell and Tissue Research 13(1): 3557-3563 (2013)

Analysis of maize inbred lines for their response to somatic embryogenesis

Dhillon, N.K. and  Gosal, S.S.

Punjab Agricultural University, Department of Plant Breeding and Genetics, Ludhiana 141 004, Punjab.
 E-mail: dhillon.navjot@gmail.com

Abstract: Screening of five maize inbred lines for their ability to form somatic embryos done. The immature embryos were used as explants for callus induction. Embryogenic calli were induced on MS supplemented with ten different concentrations of growth hormones, 2, 4-Dichlorophenoxyacetic acid, 1-naphthalene acetic acid, 6-benzylamino purine, kinetin, picloram and sucrose solidified with agar. LM 13 showed highest callus induction and subsequent embryogenesis, although the results were statistically non-significant. The somatic embryos obtained were further subcultured on to regeneration medium for studying the regeneration potential of the inbreds.

Key words: Somatic embryos, Immature embryos, embryogenesis, Inbred lines


25. Journal of  Cell and Tissue Research 13(1): 3565-3569 (2013)

Callus induction and somatic embryogenesis from maize mature embryos (Zea mays L.)

Dhillon, N.K.  and  Gosal, S.S.

Department of Plant Breeding and Genetics, Punjab Agricultural University, Ludhiana -141004.
E. mail: dhillon.navjot@gmail.com

Abstract: Ten maize lines prevalently used by breeders in the maize program were screened for their ability to form type II callus. Mature seeds were used as explant for in vitro culturing. Inbred lines showed high embryogenic response and formed friable, type II calli.  And also exhibited successful plant regeneration. Various culture media compositions based on Murashige and Skoog (1962) salts thus  investigated differed with respect to the combinations of concentrations of  auxins (picloram, 2, 4-D) and cytokinins (kinetin). Highest percent callus induction and subsequent somatic embryogenesis were observed on MS medium supplemented with picloram. The embryogenic calli were maintained and multiplied on medium containing reduced levels of auxins. Embryogenesis was characteristed, using callus morphology such as colour/nodulation and scanning electron microscopy. This inbred is considered suitable for further evaluation using a transformation system

Key words: Maize, Mature seed, Somatic embryogenesis


26.  Journal of  Cell and Tissue Research 13(1): 3571-3576  (2013)

Histology of somatic embryos from maize embryo cultures

Dhillon, N.K. and  Gosal, S.S.

Department of  Plant Breeding and Genetics, Punjab Agricultural University, Ludhiana 141004.
E mail: dhillon.navjot@gmail.com

Abstract: The histology of somatic embryo (= embryoid) formation in cultured immature embryos maize inbred (Zea rnays L.) is described. Maize embryogenic calli were obtained from immature embryos culture on Murashige & Skoog (1962) medium supplemented with Picloram (5.0 mgL-1) + 2,4-D (1.5 mgL-1) Histological examination of somatic embryogenesis revealed that somatic embryo development began with a cluster of proembryogenic cells in the peripheral region of the calli. Small and isodiametric cells with a dense cytoplasm and a prominent nucleus and nucleolus were formed on the outer cell layer of embryogenic calli; whereas the inner cell layer is composed of large cells with small nucleus and large vacuole.

Key words: Somatic embryogenesis, Maize, Histology


27.    Journal of  Cell and Tissue Research 13(1): 3577-3580  (2013)

Pathological studies on the effect of garlic  extract in acetaminophen induced liver damage in rats

Srikanth, M., Satyanarayana, M.L., Manjunatha, K. P., Rao, S. and Mallikarjuna, G.

Department of Pathology, Veterinary college, Hebbal, KVAFSU, Bangalore 560 024
E. mail: srikanthammaji@gmail.com  Telephone: 09964258029

Abstract: Garlic (well ripened) extract has been shown to protect acetaminophen induced liver injury through its antioxidant property in rats. Acetaminophen (paracetamol) is found to be a classical hepato-toxicant at the dose of 500 mg/kg. In the present study mature garlic extract was used at the dose of 200 mg/kg orally against acetaminophen induced liver injury. Garlic extract was given orally to rats for a period of 28 days. Liver damage was induced by inoculating acetaminophen at 500 mg/kg on 12th day of the experiment. Study shows centri-lobular necrosis of hepatocytes in acetaminophen treated rats.  However, when garlic extract was given along with acetaminophen the hepatic parenchyma was apparently normal indicating the beneficial effect of garlic extract in protecting liver against acetaminophen induced injury.

Key words: Garlic extract, Acetaminophen


28.    Journal of  Cell and Tissue Research 13(1): 3581-3584  (2013)

Callus culture and organogenesis in strawberry cv. Senga sengana

Jan, A.,   Bhat, K. M., Mir, M. A.,  Bhat, S. J. A. and Hassan, S.

Division of Fruit Science; Sher-e-Kashmir University of Agricultural Science and
Technology, Kashmir. 191121.  E. mail: javaidahmad44@yahoo.com

Abstract: A protocol for callus induction and regeneration of complete plantlets from leaf explants have been developed in strawberry cv. Senga Sengana. In vitro leaf explants cultured on MS medium supplemented with different concentrations of NAA, 2, 4-D alone and in combination with BAP  were kept in dark for 3 weeks.  MS medium supplemented with NAA (4.00 mg l-1) in combination with BAP (1.00 mg l-1) resulted in highest percentage (66.66%) of callus induction. Callus cultures were kept in growth chamber for 4±1 weeks on same medium for multiplication. Multiplied callus cultures were kept on regeneration medium containing different concentrations of BAP alone and in combination with NAA. Maximum shoot regeneration percentage (60.00 %) and average number of shoots (14) was obtained in MS medium supplemented with BAP (2.00 mg l-1) plus NAA (0.50 mg l-1). Rooting in regenerated microshoots was achieved in the MS half   medium containing IBA (1.00 mg l-1) and activated charcoal (100 mg l-1).

Key words: Strawberry, Callus culture


29.    Journal of  Cell and Tissue Research 13(1): 3585-3590  (2013)

Ameliorative effect of Aloe vera against beryllium induced hepatorenal toxicity

Agrawal, N. D., Agrawal, K., Shukla, S., Mathur, R. and  Rai, M.

Reproductive Biology and Toxicology Laboratory, School of  Studies in Zoology, Jiwaji University,
Gwalior 474 011 (M.P).  E. mail: agrawalnarottam@gmail.com  

Abstract: Aloe vera (50, 100, 150 and 200mg/kg p.o.) was screened to encounter the characteristic biochemical alterations followed by subchronic exposure to beryllium (1mg/kg i.p. once a day for 35 days). Exposure to beryllium nitrate disturbed the carbohydrate metabolism by significantly decreased the level of blood sugar, glycogen and the activity of G-6-Pase in liver. Beryllium nitrate significantly elevated leakage of ALT, AST, urea, bilirubin and creatinine in serum, decreased hemoglobin and induced oxidative stress resulting in elevation of lipid peroxidation and reduction in reduced glutathione. A significant fall was observed in tissue protein, activitities of ALPase, ATPase in liver and kidney due to beryllium intoxication. The different doses of Aloe vera reversed the disturbed carbohydrate metabolism and the alterations of other variables more towards control. It was concluded that dose of 150 mg /kg of Aloe vera was found to be most effective in attenuating beryllium induced hepatorenal toxicity in rats.
         
Key words: Beryllium toxicity, Aloe vera


30. Journal of  Cell and Tissue Research 13(1): 3591-3596  (2013)

Interleukin-8 gene polymorphism and its association with mastitis in Murrah buffalo

Sharma, R., Sangwan, M.L., Singh, P. and  Rana, A.

Department of Animal Biotechnology, L L R University of Veterinary and Animal Sciences,
Hisar  125004, Haryana. E. mail: raulsikandar@gmail.com Cell: 09697025323

Abstract: Present study was carried out on Interleukin-8 gene of Murrah buffalo with an objective to find out different variants of Interleukin-8 gene and its association with mastitis. Genomic DNA was isolated from randomly selected lactating buffalo maintained at from Govt. Livestock Farm, Hisar; Central Institute for Research on Buffaloes, Hisar, Buffalo Research Centre, College of Animal Sciences and Veterinary clinics, CCSHAU, Hisar by phenol-chloroform method. Quality and quantity of DNA was checked and it was found to be of good quality as determined by agarose gel electrophoresis. A set of primers was designed by using Fast-PCR and primer 3 software covering 853 bp nucleotides of IL-8 gene which was reported highly polymorphic in previous studies. PCR products of 853 bp were obtained. PCR-RFLP was done for all the samples from healthy and mastitis animals using HaeIII and HinfI restriction enzymes. Pattern of resolution of digested fragments by both the restriction enzymes was found same in both two groups of animals. All monomorphic bands were found. Target sequence IL-8 gene of Murrah buffalo was sequenced which has been found to span over 754 bp long. This sequence was compared to the sequence of Bubalus bubalis sequence at NCBI. Multiple alignments of IL-8 target sequences of Bubalus bubalis between two groups have revealed no variation. Basic Local Alignment Search Tool (BLAST) analysis revealed sequence identity of target region (IL-8 gene) of Murrah buffalo with Bubalus bubalis sequence at NCBI was 98 to 100%, 90 to 95% with Bos taurus, 90 to 97% with Bos indicus and 89-92% with Ovies aries.

Key words: Murrah buffalo, Interleukin-8, RFLP, BLAST, HaeIII, HinfI

31.    Journal of  Cell and Tissue Research 13(1): 3597-3601 (2013)

Anti-diabetic effects of Withania somnifera  root  and  leaf extracts  on  streptozotocin induced diabetic rats

Sarangi, A., Jena, S., Sarangi, A.K. and  Swain, B.

P. G. Department of  Botany, Utkal University Bhubaneswar  751024, Odisha.
E. mail: anshumanbbsr2@gmail.com

Abstract:The present investigation explores the possibilities of using the root and leaf extracts of an important plant Withania somnifera targeting DM and to examine their hypoglycaemic and hypolipidaemic effects on Streptozotocin-induced diabetic rats. Withania somnifera root extracts (WSREt)  and leaf extracts (WSLEt) were orally administered daily to diabetic rats for eight weeks. After the treatment period, blood glucose and serum enzymes like aspartate transaminase (AST), alanine transaminase (ALT) and alkaline phosphatase (ALP), total cholesterol, triglycerides, HDL-c high density lipoprotein-bound cholesterol, LDL-c low density lipoprotein-bound cholesterol l, LDH and serum proteins levels were determined. The levels of blood glucose, AST, ALT,  ALP, LDH, serum lipids except high density lipoprotein-bound cholesterol (HDL-c) were significantly increased, but total protein albumin, albumin : globulin (A : G) ratio, were significantly decreased in streptozotocin-induced diabetic rats. Treatment of the diabetic rats with root and leaf extracts  altered the changes of the above parameters and restored them after eight weeks of treatment, indicating that the extracts possess hypoglycaemic and hypolipidaemic properties, hence useful in  diabetes mellitus.

Key words:  Diabetes mellitus, Withania somnifera, Streptozotocin

 
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