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Tissue Research » Past Abstracts
1. Journal of  Cell and Tissue Research 13(2): 3603-3609  (2013)
 
 
Human platelet lysate as a substitute for fetal bovine serum in human keratinocyte cell cultures conforming epithelia for transplantation
 
Altran, S.C., Yoshito, D., Isaac, C., Sufi, B.S., Herson, M.R., Conceição, R.O., Ferreira, M.C.  and Mathor, M B.
 
 Radiation Technology Center (CTR),Nuclear and Energy Research Institute, IPEN – CNEN/SP, Brazil
 E. mail: silaltran@yahoo.com.br
 
Abstract: Confluent epithelia of cultured human autologous keratinocytes have been transplanted in several critical clinical situations. However, the presence of xenobiotic components, in particular of fetal bovine serum in the cell culture media as a nutritional complement, introduces a risk to this practice. This study evaluates human platelet lysate in different concentrations, used to replace fetal bovine serum in primary keratinocyte plating and further cell culture maintenance, where the rate of cell adhesion and viability were used as parameters. Sodium heparin was added at different moments of the lysate preparation in order to avoid the coagulation of the complete culture medium. Filtration to obtain a homogeneous and sterile product was a cumbersome process, wherethe best option was also established based on the obtained supernatant volume, the quantity and the quality of suspended material and presence of cell pellet and/or surface aggregates. Overall results allow for the conclusion that human platelet lysate, to which heparin is added before the sterilization filtration steps can best replace fetal bovine serum in keratinocyte culture media when used in the concentration of 2.5% for cell plating and 2.5% or 5% for cell culture maintenance.
 
Key words: Platelet lysate, Human cell culture, Wound healing
 
2. Journal of Cell and Tissue Research 13(2): 3611-3617 (2013)
 
 
Molecular characterization as a significant tool for identification of Indian puffer fishes
 
Badhe, M.R. and Nandanpawar, P.C. 
 
Department of Fish Genetics and Biotechnology division, Central Institute of Fisheries Education,
Versova, Mumbai, 400 061. E. mail: mohanbadhe1212@gmail.com        .
                                                                  
Abstract: Biodiversity characterization is essential for sustainable utilization of resources and defining biological entities for conservation. India has blessed with rich biodiversity but few species have been formally documented till now. Due to the inherent problems with traditional taxonomy, DNA barcoding, i.e. the use of 655bp region of mitochondrial gene cytochrome c oxidase I (COI) has recently been promoted as efficient method for species-level identifications, biodiversity characterization and discovery of species. The present study was taken up to develop DNA barcodes for 3 species of puffer fishes. 15 specimens of puffer fishes were collected from various parts of India such Mumbai, Ratnagiri, Mandapam and Chennai. The specimens were identified by traditional taxonomical tools and then were preserved in 100% ethanol for further analysis. Specific primers were used to amplify the 680bp partial cytochrome C oxidase sub unit 1 (COI) gene region. The cytochrome C oxidase sub unit 1 barcode sequence clearly distinguished all the species with high percentage of interspecific distance than intra specific distance. The average interspecific distances were 0.20 between species of puffer fishes. In puffer fishes, the average transversional pairs were relatively more than average transitions. The overall transition and transversional (Ti/Tv) rate bias for cephalopods were 2.025. The neighbor-joining tree revealed distinct clusters shared by the species of same genera.
 
Key words:DNA Barcoding, puffe rfishes, Cytochrome C oxidase I
 
3, Journal of Cell and Tissue Research 13(2): 3619-3624 (2013)
 
Chitosan nanoparticles for gene delivery in Macrobrachium rosenbergii (De Man 1879)
 
Nandanpawar, P.,  Badhe, M., Rather, M. A. and Sharma, R.
 
Department of Fish Genetics and Biotechnology division, Central Institute of Fisheries Education,
Versova, Mumbai, 400 061. E. mail: mohanbadhe1212@gmail.com
 
Abstract: The present study demonstrates non viral gene delivery using chitosan nanoparticles in Macrobrachium rosenbergii as model organism. Chitosan nanoparticles were synthesized by ionic gelation method. The mean particle size of chitosan nanoparticles was 27.6nm. The Polydispersity index was found to be 0.448 and zeta potential was +24.79mV. Green fluorescent protein encoding plasmid (PDB-402) was used for conjugation with these nanoparticles. Chitosan (nanoparticle concentration: 40mg/L) - DNA nanoconjugates (plasmid concentration: 50ng/µl) were administered to prawns via immersion treatment for a period of 36 hrs. The gene expression was studied using PCR and fluorescence microscopy which showed green fluorescent protein expression in muscle, gills and pleopods indicating wide tissue distribution. This study also affirms that immersion treatment can be efficiently employed for DNA- nanoconjugate delivery as it minimises the stress in animals.
 
Key words:Chitosan nanoparticles, Gene Delivery, Macrobrachium rosenbergi
 
4      Journal of Cell and Tissue Research 13(2): 3625-3630 (2013)
 
Effects of cryoprotectants on freezing, culture behaviour of buffalo umbilical cord matrix cells
 
Singh, P., Rose, M.K., Yadav, P.S., Sharma, R. and Singh, J.
 
Department of Veterinary Physiology and Biochemistry, College of Veterinary Sciences,
Lala Lajpat Rai University of Veterinary and Animal Sciences, Hisar 125004. Haryana
E. mail: preeti0829@gamil.com Cell 09697025323
 
Abstract: Umbilical cord represents the link between mother and fetus during pregnancy. Postnatally umbilical cord is a discarded organ and the collections of umbilical cord cells do not require an invasive procedure with ethical concerns. It is composed of a special embryonic mucous connective tissue called Whartson’s jelly and its cells have the properties of stem cells. Umbilical cord matrix cells are cryopreserved to study their future applications. Cryopreservation is a method to protect cells from freezing injury. In the present study confluent cultures from buffalo umbilical cord matix cells were frozen at three different passages (P3, P6, P9) in three different combinations of cryoprotectants viz. DMEMculture medium containing 20% FBS and 10% DMSO,   1.5 M ethylene glycol and 0.2 M sucrose in PBS and 4% DMSO, 6% trehalose and 90% FBS. Pre and post freezing cell viability (%) was assessed using trypan blue dye exclusion method. The cells in (FBS-DMSO) and (EG-Sucrose) had more post thaw viability than (D text-autospace:none"> 
Key words: Cryoprotectant, Stem cell, Umbilical cord
 
5      Journal of Cell and Tissue Research 13(2): 3631-3638 (2013)
 
Preclinical safety assessment of silk protein derived biomaterial in laboratory animals
 
Padol, A.R.,Jayakumar, K., Shridhar, N.B., Mohan, K. and Manochaya, S.
 
Department of Pharmacology and Toxicology, Veterinary College, KVAFSU, Hebbal, Bangalore (560 024). E. mail: dramolpadol@gmail.com
 
Abstract: Silk proteins, fibroin and sericin have been extensively used as biomaterial in the management of tissue injuries. We have previously evaluated the novel silk protein based membranes (SP-membranes) for full skin thickness wound management. In the present study, we have evaluated the preclinical safety of the SP-membrane in the animal models to obtain its toxicological profile. For safety assessment of the SP-membrane, the acute toxicity test was performed in rats, dermal irritation study in new Zealand white rabbits and skin sensitization potential was assessed in guinea pigs according to Buehler test method. There was no mortality or clinical signs observed in the rats dermally treated with SP-membranes. Also there were no significant changes in body weight and serum biochemical parameters in the acute dermal toxicity. In acute dermal irritation study, the score of skin reaction in treated rabbits was “0” for all time points of observations; hence the SP-membrane was classified as non-irritant to skin. No skin reactions in terms of erythema and edema were observed in the treated guinea pigs in skin sensitization study at 24 and 48 h of challenge patch removal, which was scored “0” based on the Magnusson/Kligman grading scale and SP-membrane was considered as non-sensitizing to guinea pigs. The data obtained in this investigation provides powerful support for safety and biomedical application of SP-membrane in human therapeutics.
 
Key words: Biomaterial, Preclinical safety, Silk protein
 
6      Journal of Cell and Tissue Research 13(2): 3639-3642 (2013)
 
Diagnostic accuracy of creatine kinase muscle brain isoenzyme (ck-mb) in various cardiac diseases in dogs
 
Rajkumar, K., Ansar Kamran, C., Upendra, H.A. and Yathiraj, S.
 
Department of Veterinary Medicine, Veterinary College, KVAFSU, Hebbal, Bangalore 560024,
Email: rajvet10@gmail.com Mobile +918861638285
 
Abstract: Present research was undertaken to evaluate diagnostic accuracy of creatine kinase muscle brain isoenzyme (CK-MB) in various cardiac diseases in dogs. Dogs presented to the cardiology service of the Veterinary Teaching Hospital, Veterinary College Bangalore from October 2011 to September 2012 were eligible for entry into the study. All dogs were evaluated based on physical examination, radiography, electrocardiogram and echocardiography. CK-MB was estimated using semi-automatic biochemical analyser using commercial reagents. In the present study CK-MB values were more than 6.3 IU/L in 98.4 per cent of cases. In conclusion, CK-MB is not 100% specific for the heart. In presence of clinical signs like inappetence, coughing, ascites, dyspnoea, exercise intolerance and jugular pulsation elevated CK-MB suggests that there was a myocardial damage.
 
Key words: Createnine kinase, Cardiac disease
 
7       Journal of Cell and Tissue Research 13(2): 3643-3646 (2013)
 
Diagnostic accuracy of lactate dehydrogenase in various cardiac diseases in dogs
 
Rajkumar, K. and Ansar Kamran, C.
 
Department of Veterinary Medicine, Veterinary College, KVAFSU, Hebbal, Bangalore 560024
E. mail: rajvet10@gmail.com Mobile +918861638285
 
Abstract: The study was undertaken to evaluate diagnostic accuracy of lactate dehydrogenase (LDH) in various cardiac diseases in dogs. Dogs presented to the cardiology service of the Veterinary Teaching Hospital, Veterinary College Bangalore from October 2011 to September 2012 were eligible for entry into the study. All dogs were evaluated based on physical examination, radiography, electrocardiogram and echocardiography. LDH was estimated using semi-automatic biochemical analyser using commercial reagents. LDH values were less than 50 IU/L in 22 per cent, 50-380 IU/L in 45.1 per cent and more than 380 IU/L in 32.9 per cent of the study samples and expected range was between 23.72-43.66 at 95 per cent confidence interval. In the present study only thirty three per cent of the cases showed increased lactate dehydrogenase (LDH) concentration in blood. However, skeletal muscle injury can also result in increased in serum LDH values. In the absence of muscle injury elevated LDH concentration in the blood act as a sensitive indicator of myocardial cell damage, but our present study conclude that its sensitivity was low. New generation cardiac biochemical markers which are more specific for myocardial cell damage like cardiac troponin I and prohormone pro B-type natriuretic peptide are recommended for diagnosis of cardiac disease in dogs.
 
Key words: Lactate dehydrogenase, dogs, cardiac disease
 
8. Journal of Cell and Tissue Research 13(2): 3647-3650 (2013)
 
Lack of association between TLR2 gene and mastitis in Murrah buffaloes
 
Rana, V.P., Sangwan, M.L., Kumar, A., Verma, V., Kumar, V. and Khirbat, R.
 
Department of Animal Biotechnology, Lala Lajpat Rai University of Veterinary and Animal Sciences,
 Hisar 125004, Haryana. E. mail: virenderr@gmail.com, Cell 09468426467
 
Abstract: In the present study, the buffalo TLR2 gene was taken as a candidate gene for association study with mastitis resistance. Blood samples of 46 Murrah buffaloes were taken in which 23 animals were suffering from mastitis. The samples were collected from Buffalo Research Centre, TVCC, LLRUVAS, and local organized farms in Hisar. The genomic DNA was isolated by phenol chloroform method. PCR was standardized to amplify partial exon 2 (448 bp) of TLR2 gene using published primers [1]. PCR-RFLP analysis of the partial exon 2 region of TLR2 gene was studied using Dra I, Eae I and Hph I restriction endonucleases which gave restriction products of 300bp, 148bp; 317bp, 131bp; and 200bp, 148bp, 100bp, respectively, in all the animal under study. Hence role of exon 2 of TLR2 region with mastitis cannot be established.
 
Key words: TLR2 gene, Murrah buffalo
 
9. Journal of Cell and Tissue Research 13(2): 3651-3656 (2013)
 
Clinico-pathological and pathomorphological changes induced by Mycoplasma mycoides sub sp. Mycoides large colony in Wistar albino rats
 
Vijayalakshmi, T.C., Narayanswamy, H.D., Satyanarayana, M.L., Ayoub, S.M., Krishnaveni, N., Gajendragad, M., Uma, S., Suryanarayana, T. and Enakshi, G.
 
Veterinary College Hebbal, Bangalore, KVAFSU, Bidar. E. mail: mudasir581@gmail.com
 
Abstract: The present study was carried out to study the pathomorphological changes and haematological alterations induced by Mycoplasma mycoides subsp. mycoides large colony in wistar albino rats for 45 days. Thirty wistar albino rats in each group were administered with 108, 109 and1010 CFU/ml MmmLC suspension intranasally respectively, with phosphate buffered saline (PBS) control group. It was observed that Mmm LC suspension inoculated group showed pyrexia, dullness, depression, reduced feed intake, sneezing and serous nasal discharge. There was no significant difference between mycoplasma infected and normal control group with respect to TLC, DLC and Hb except for neutrophils. Pathomorphological lesions induced by MmmLC suspension were of congestion, haemorrhages,consolidation of lungs and thickening of synovial membrane and were concentration dependent.Trachea, lungs and joints(elbow, stifle joints) were the most affected organs of MmmLC infected in rats.
 
Key words: Mycoplasma mycoides, Pathomorphological changes
 
10 Journal of Cell and Tissue Research 13(2): 3657-3661 (2013)
 
Comparative study of surgical excision and chemot-herapeutic agents in the treatment of mammary tumours in canine
 
Talekar, S.H., Gahlod, B.M., Patil, S.N., Dakate, M.S. and Bhandarkar, A.G.
 
Department of Surgery and Radiology, College of Veterinary Science and Animal Husbandry, JAU,
Junagadh 362001. E.mail: shivaji.talekar@gmail.com
 
Abstract: Eighteen female dogs with mammary tumor were included in this study. Animals were randomly divided into three groups of six each. Surgical excision of mammary tumour was carried out in group I. Group II received an intravenous injection of 30mg/m2 doxorubicin hydrochloride twice at an interval of 21 days, whereas 0.025mg/kg vincristine sulphate was given to group III thrice at weekly interval. Clinical and haematological observations were recorded on 0, 1, 3, 7, 14, 21, 22 and 28 day. Representative samples of tumor mass were collected from all the animals for histological analysis. In group I, recurrence of tumor was observed in only one case. Complete regression of tumor was observed in four animals of group II and only one animal of group III. Mean body temperature was significantly higher in group I as compared to group II and III. Variations in mean heart rate were statistically non-significant. Increase in mean respiration rate was found to be significant either among periods or among groups. Mean total erythrocyte count (TEC) value showed significant variation among groups. Variation in mean total erythrocyte count (TLC) value was significant among groups. There were non-significant changes in mean haemoglobin and packed cell volume (PCV) value either within or among groups. Histopathological observations confirmed that 66.66% tumor was of malignant type and 33.33% of benign type. From the present study, it may be concluded that surgical excision of mammary tumor was found to be is more effective followed by doxorubicin hydrochloride for the treatment of mammary tumor in canine.
 
Key words: Mammary tumor
 
11. Journal of Cell and Tissue Research 13(2): 3663-3667 (2013)
 
Evaluation of dual drug eluting stent (heparin and sirolimus) into renal artery using retrograde trans-femoral canine model
 
Talekar, S.H., Patil, D.B.,Parikh, P.V.,Mathews, A., Rajapurkar, M.M., Patil P.B. and Kelawala, N.H.
 
Department of Surgery and Radiology, College of Veterinary Science and Animal Husbandry, JAU, Junagadh – 362001. E. mail: shivaji.talekar@gmail.com Cell: 09833149004
 
Abstract: In the present study, a new concept of dual DES (HSES) with biodegradable polymer was evaluated for ISR {due to increased neointimal hyperplasia (NH), peristrut inflammatory response (PIR), lumen narrowing (LN), non onstructive peristrut fibrin (NOPF), internal elastic lamina disruption (IELD) and delayed apparent reendothelization (AR)} in balloon injured renal polar artery model of dogs (n=12) which were randomly bred, apparently healthy mongrel breed, weighing 15kg (avg.). Out of 12 mongrel dogs, HSES and SES were deployed at right renal arteries in 6 dogs each, which, further sub-grouped on stent retrieval period (group A - 30 days and group B - 90 days) {i.e. HSES-A (n=3), HSES-B (n=3), SES-A (n=3) and SES-B (n=3)}. Analyzing the results, it was found that both the stents altered the renal function parameters non-significantly. Thus assessing the renal tolerability both of the stents can be graded equivalently except urinary enzymes where group SES showed significant alterations, while, HSES group showed superior histological alterations at 30 and 90 days occlusion study as compared with SES. Therefore, the use of HSES can be preferred in patients with arterial stenosis renal dysfunction.
 
Key words: Heparin, Sirolimus, Renal artery
 
12 Journal of Cell and Tissue Research 13(2): 3669-3672 (2013)
 
Guidelines for histological analyses of vascular stent in animal models
 
Talekar, S.H., Patil, P.B., Patil, D.B., Mathew, A., Parikh, P.V.,Kelawala, N.H., Rajapurkar, M.M. and Lele, S.S. 
 
Department of Surgery and Radiology, College of Veterinary Science and Animal Husbandry, JAU, Junagadh 362 001. E. mail: shivaji.talekar@gmail.com
 
Abstract: Interventional surgery has been a breakthrough for all obstructive disorders in hollow tissues. Stents are most commonly used devices, which are often delivered using catheter guided systems. Biomedical models in standardizing or evaluating stent materials and mechanics have been increased tremendously over the last few decades. Histological analysis has been the most commonly used method for concluding results or evaluating material. Among many biomedical animal models, we have used rabbit and dog, which were evaluated on few methods. On the basis of few years experience, in the field of stenting, we mentioned few guidelines to perform evaluation for such experiments with ease and efficacy.
 
Key words: Resin embedding, Vascular stent, Histomorphometry
 
13. Journal of Cell and Tissue Research 13(2): 3673-3677 (2013)
 
Evaluation of heparin-sirolimus eluting stent (hses) with sirolimus eluting stent (ses) for anti-thrombotic properties in external iliac arteries of rabbit
 
Patil, P.B., Patil, D.B.,Talekar, S.H., Parikh, P.V., Mathews, A., Lele, S.S., Rajapurkar, M.M. and Kelawala, N.H.
 
Department of Surgery and Radiology, AAU, Anand 388001. E. mail: shivaji.talekar@gmail.com
 
Abstract: In the present study, randomly bred, 21 apparently healthy NZW adult rabbits of either sex, with average body wt of 2.168 kg were used for comparative evaluation of HSES with SES1 in bilateral external iliac arteries for SAT and ISR. In 21 NZW rabbits, HSES and SES were deployed at left and right external iliac arteries, respectively using millennium matrix delivery system. Each group was allotted to three sub-groups VIZ., A (7 days, n=7), B (14 days, n=7) and C (21 days, n=7) and evaluated for increased neo-intimal hyperplasia (NH), peristrut inflammatory response (PIR), luminal narrowing (LN), non obstructive peristrut fibrin (NOPF), internal elastic lamina disruption (IELD) and delayed apparent re-endothelisation (AR) to study ISR. On histological examination, IELD was clearly and prominently seen than the external elastic lamina in rabbit iliac artery, similar to that of the human artery. On comparison with control (SESs), arteries with HSES showed marked reduction {11 out of 21 (52.29%) with SES versus 3 out of 21 (14.29%) with HSES} in non-occlusive peristrut fibrin deposition. Further, concomitant reduction in neointimal hyperplasia {38% versus 80% in SES}, peri-strut inflammatory response {19% versus 66% in SES}, lumen narrowing {9.52% versus 52.38% in SES} and early complete re-endotheliazation and intimal healing {14.29% versus 42.86% IELD in SES} establishes superiority of HSES over SES in prevention of stent thrombosis and ISR. On comparison with control (SESs), arteries with HSES showed superiority histological endpoints in percentage viz., NOPF, NH, PIR, LN, AR (14, 38, 19, 10, 90, 14)% compared with SES (52, 80,66, 52, 90, 43)% respectively.
 
Key words: External iliac arteries, HSES, SES, Heparin-coated stents
 
14. Journal of Cell and Tissue Research 13(2): 3679-3682 (2013)
 
Histomorphology of hair follicle pattern in wild big cats and sloth bear skin: A tool for forensic identification
 
Shambhulingappa, Y.B., Prasad, R.V., Jamuna, K.V. and Ramkrishna, V.
 
Department of Veterinary Anatomy and Histology, Veterinary College, Hebbal, Bangalore - 560 024,
E. mail: rvprashwin@gmail.com, Cell 09449855893
 
Abstract: This study comprised of horizontal skin sections obtained from big cats and sloth bear. The Leopard skin showed a single primary hair follicle surrounded by circumferentially arranged groups of compound follicles. Each compound follicle consisted of 6-8 fine secondary hair follicles interspersed with fine connective tissue fibers. Bengal tiger skin showed sparsely distributed compound follicles. Within each compound follicle a primary hair follicle located at one end underneath which 6-8 secondary hair follicles were observed. Lion skin comprised of clusters of compound hair follicles around a primary hair follicle bundle. Within each compound follicle a primary hair follicle was always noticed. In sloth bear skin sparsely distributed elliptical shaped compound hair follicles were recorded. Within each compound follicle 2-4 primary hair follicles were irregularly arranged. Thus on the bases of hair follicle structure of skins various wild animals can be identified.
 
Key words: Wild animals, Skin, Hair follicles
 
15. Journal of Cell and Tissue Research 13(2): 3683-3686 (2013)
 
Fine morphology of interstitial gland tissue in normal cycling goat (Capra hircus) ovary
 
Sharma, R. K. and Saini, S.
 
Reproductive Physiology Laboratory, Department of Zoology, Kurukshetra University,
Kurukshetra 136119. E. mail: sdhsaini@yahoo.co.in
 
Abstract: The fine structure of interstitial gland cells (IGCs) of mature normal cycling goat ovary was analyzed. The IG cells were found to be present in clusters or rows surrounding the developing and atretic follicles, corpus luteum and corpus albicans. During electron microscopic studies these cells were distinguished by their glandular appearance, large size and prominent nucleus. IGCs were characterized by large voluminous nucleus having indented to lobbed contour. Nucleus possessed a darkly stained eccentrically placed prominent nucleolus. The cytoplasm revealed the presence of a large aggregations comprised of different cell organelles such as Golgi complex, spherical mitochondria, pleiomorphic bodies, abundant smooth endoplasmic reticulum, glycogen granules and a numerous lipid droplets. The relative percentile of smooth endoplasmic reticulum was found to be highest (16.14%) followed by mitochondria (4.63%), lipid droplets (2.57%) and Golgi bodies (1.1%). The ultrastructural analysis and the relative abundance of the cell organelles observed in interstitial gland cells clearly indicate the steroidogenic potential of the interstitial gland cells. It can be suggested from the present study that the interstitial gland cells have role in the hormone production in goat ovary as they possess the steroidogenic morphological characters.
 
Key words: Goat, Ovary, Interstitial gland cells
 
16.   Journal of Cell and Tissue Research 13(2): 3687-3690 (2013)
 
Evaluation of antimicrobialand antifungal activities ofsome newly synthesized 2,4-disubstituted-1,5-benzodiazepines
 
 Salve, P.S. and Mali, D.S.
 
Department of Pharmaceutical Chemistry, KLE University’s College of Pharmacy, Belgaum
 590 010, Karnataka. E. mail: preeti.salve@gmail.com Cell: 09673834221
 
Abstract: In  present study the antimicrobial activity of a new series of 2,4-disubstituted-1,5-benzodiazepine derivatives through an eco-friendly synthesis are reported. This was achieved by the condensation of o-phenylenediamine and various substituted chalcones under microwave irradiation. The structures of the synthesized compounds were confirmed through IR and 1H NMR spectroscopy. The compounds were evaluated for their antibacterial and antifungal activities by using minimum inhibitory concentration (MIC) method. Synthesized compounds were tested against four strains of bacteria viz. Escherichia coli, Enterococcus faecalis, Pseudomonas aeruginosa, Staphylococcus aureus and two strains of fungi viz. Candida albicans, Aspergillus niger. Brain heart infusion (BHI) broth was used as nutrient medium. Ciprofloxacin and fluconazole were taken as standard drugs in antibacterial and antifungal activity, respectively. All the synthesized compounds showed moderate to good antibacterial and antifungal activities. Compounds 5b, 5c, 5d, 5e and 5h have shown significant antifungal activity against Aspergillus niger at a very low concentration of 0.4 µg/ml. Compounds 5b and 5c have shown significant antibacterial activity against Enterococcus faecalis at a low concentration of 3.12 µg/ml.
 
Key words: 1,5-benzodiazepines, Antibacterial activity, Antifungal activity
 
17. Journal of Cell and Tissue Research 13(2): 3691-3696 (2013)
 
Isolation and characterization of collagen from prawn (Penaeus indicus)
 
Shah, V. and Manekar, A P.
 
Department of Zoology, Institute of Science, Mumbai 400 032, India.
E. mail: ashok.manekar@rediffmail.com
 
Abstract: Prawn (Penaeus indicus) has been used as food for human consumption and valuable products having potential nutraceutical and pharmaceutical applications. In the present investigation, collagen from the prawn (Penaeus indicus) was isolated and characterized. Fourier transform infrared spectroscopy revealed that prawn tissue collagen molecules had the compact triple helical structure stabilized mainly by the hydrogen bond. The prawn tissue collagens were classified as type I collagen containing α1 chains and one α2 chains with no disulfide bond. To check the potential of the prawn tissue collagen,, human foreskin fibroblast (HFF) were cultured on prawn tissue collagen and their growth was evaluated by MTT method. We observed steady growth on placenta collagen. CD44 protein expression were observed by staining on HFF grown on prawn tissue collagen as well as vimentin compared to tissue culture dishes was evaluated by immunoflurosence as well as by Fluorescence-activated cell sorting (FACS) analysis. It is inferred that the collagen isolated from prawn (Penaeus indicus) has potential for use as an alternative to collagen in food and pharmaceutical industries.
 
Key words: Collagen, Prawn
 
18. Journal of Cell and Tissue Research 13(2): 3697-3699 (2013)
 
Studies on larval gut lipolytic activity in Helicoverpa armigera (hubner)
 
Pawar, A.S.,  Gejage, R.M. and Dubal, R.S.
 
Dept. of Zoology, Shri Vijaysinha Yadav Arts and Science College, Pethvadgaon, Tal. Hatkanangale, Dist. Kolhapur-416 112 (M.S).   E. mail: saishwaryapawar@yahoo.in
 
Abstract: Larval gut lipolytic activity in gram pod borer, Helicoverpa armigera (Hubner) revealed optimum pH 7.7, incubation time 20 minutes, temperature 370C, enzyme concentration 1 % and substrate concentration 6 % and Michaelis-Menten constant 0.129×10-2 mM. The gradual increase in larval gut lipaseactivity was observed from 7-day larvae to 10-day larvae and decrease from 10-day larvae to 15-day larvae. The maximum lipase activity was observed in 10-day larvae. The physiological role of larval gut lipase (EC 3.1.1.3) in H. armigera (Hubner) has been reported in present paper.
 
Key wordsLipase, Helicoverpa armigera, Larva gut
 
19. Journal of Cell and Tissue Research 13(2): 3701-3709 (2013)
 
Cloning of drought related genes in vigna aconitifolia through modified differential display
 
Soni, P., Mohapatra, T., Bhatt, K. V., Singh, G., Rizwan, M. and Sharma, R.
 
Plant Biotechnology centre, Swami Keshwanand Rajasthan Agricultural University, Bikaner 334 003,
Rajsthan. E. mail: priyasoni28@gmail.com, Cell 08447095503
 
Abstract: A total of 12 genotypes of moth bean were evaluated for desiccation tolerance at germination and seedling stage. Differential response was observed at 8% and 15% PEG on germination and seedling level respectively where Varieties RMO-435, RMM-12-Single, RMM-12-Poly and CZM-105 were found tolerant. Certain protein were found to be upregulated with treatment in all the samples while some showed upregulation only in tolerant types through SDS PAGE analysis. Twenty four primer combinations of anchored poly T (3) and random primers (8) were used to amplify the transcripts and amplicons were resolved on agarose gel with ethidium bromide. This modified DDRT-PCR assay amplified a total of 95 bands with an average of 4.04 per primer combination. 43 gene fragments were found to be up-regulated whereas 34 were down-regulated in response to drought as compared to the control. A total of 23 unigenes (4 contigs and 19 singletons), were derived by cluster assembly and sequence alignment of 29 ESTs; Out of 29 ESTs, 4 EST (13.79%) was found to be novel to moth bean. Significant BLASTX similarity (<1E-06) in the NCBI non-redundant (nr) database was detected for 25 ESTs (86.20%). Gene ontology functional classification terms (BLASTX results and GO terms), were retrieved for all sequences and 23 sequences were annotated with 8 enzyme commission (EC) codes and were mapped to 11 different KEGG pathways. Our study lists mainly the transcripts that are differentially expressed under PEG induced desiccation stress, where 9 chaperones including HSP 70, HSP 101 and Dnaj like proteins were found.
 
Key words: Moth bean. Drought. SDS PAGE
 
20. Journal of Cell and Tissue Research 13(2): 3711-3714 (2013)
 
Effect of growth hormones on in vitro mass- propagation of orchid species Rhyncostylis retusa
 
Kalita, R.,   Das, A.K., Gogoi, H.K. and Veer, V.
 
Defence Research Laboratory, Tezpur 784 001 (Assam) E. mail: ajit_drl@rediffmail.com
 
Abstract: Indehiscent mature pods of orchid species Rhyncostylis retusa from North-East India were cultured on Knudson’s culture medium for seed germination. The production of orchid seedlings from seeds involved three sequential phases – Germination, Protocorms formation and Seedling development. Best response for seed germination (75-95%) in 15 days was observed on Knudson’s medium supplemented with NAA 3.0 mg/l, BAP 5.0 mg/l and GA3 1.0 mg/l. Again, shoot initiation was recorded in 75 days and root initiation in 95 days. Complete plantlets with well defined shoot and root were cultured in half strength Knudson’s medium. After 180-190 days, in vitro plantlets were transferred in to the pots. 75-85% survival rate was recorded. The hormone concentration was different in each phase.
 
Key words: Orchid seed, In vitro propagation, Hormones
 
21. Journal of Cell and Tissue Research 13(2): 3715-3720 (2013)
 
Radical burst: A pivotal mechanism of plant bio-defense and signal transduction
 
Ahanger, R.A., Bhat, H.A., Dar, N.A., Ganie, S.A., Lone, A.A. and Shah, A.H.
 
Division of Plant Pathology, Sher-e-Kashmir University of Agricultural Sciences and Technology of Kashmir, Kashmir (J&K). E.-mail: rayees83@yahoo.in Cell: 099067 79151
 
Abstract: Plants have evolved an array of defensive tools to resist stresses, including those from attack by pathogens. One of the earliest events upon recognition of a pathogen is the accumulation of reactive oxygen species (ROS), represented by hydrogen peroxide (H2O2), superoxide (O2-) and hydroxyl radical (•OH). Biotrophs prefer living, while necrotrophs prefer dead cells for nutritional purposes and therefore, tissue necrosis caused by reactive oxygen species (ROS) during pathogen infection increases host susceptibility to necrotrophic, but resistance to biotrophic pathogen. ROS serve as a second messenger in a systemic signalling network in plant immunity and leading to the expression of defense genes, phytoalexin production, and triggering of a hypersensitive response (HR). Avirulent pathogens often induce a biphasic ROS accumulation with a small, first phase, followed by a continuous phase of much higher intensity. Nitric acid (NO) signaling also participates in defense responses against necrotrophic pathogens in a manner other than the antioxidant of ROS. A variety of enzyme systems have been implicated in ROS generation following pathogen recognition viz., (NADPH) oxidase and superoxide dismutase . Salicylic acid, jasmonic acid and ethylene can influence induction of ROS and antioxidants thereby influencing resistance or susceptibility of plants to pathogens.
 
Key words: Plant bio-defense, Signal transduction, ROS
 
22. Journal of Cell and Tissue Research 13(2): 3721-3727 (2013)
 
Genetic augmentation of kinnow through direct gene transformation with OsglyII gene construct
 
Kour, A., Rattanpal, H.S. and Sidhu, G.S.
 
Department of Fruit Science, Punjab Agricultural University, Ludhiana 141004
E. mail: amardeepkour81@gmail.com, Cell: 08427600775
 
Abstract: The present study was carried out with the objective to undertake the genetic transformation through micro projectile using Osgly II gene, capable of imparting salt tolerance. The salinity tolerance Glyoxalase II gene (OsglyII) construct from Oryza sativa with GUS and hpt with 35S promoter in plasmid pCAMBIA 1304 was used for transforming epicotyls segments. The optimum concentration of hygromyvin B was 2 mg l-1and the transient GUS expression percentage 48 h after bombardment was 22.00. The transformed epicotyls segments exhibited the selection percentage of 9.5 per cent on MS + BAP 1.0 mg l-1+ 2 mg/l hygromycin. The selected segments showed the regeneration percentage of 3.1 respectively. 13 putative transgenic plants were obtained out of 421 epicotyl segments. Histochemical GUS analysis revealed two GUS positive plants from randomly selected 14 putative Kinnow plants (14.29 per cent expression). PCR analysis revealed 3 out of 13 plants positive for Osgly II gene and the per cent transformation efficiency was 0.71.
 
Key wordsOsglyII gene, Kinnow, Transgen
 
23. Journal of Cell and Tissue Research 13(2): 3729-3737 (2013)
 
Protective effect of aqueous bark extract of Terminalia arjuna against copper-ascorbate induced oxidative stress in vitro in goat liver
 
Ghosh, A.K., Mitra, E., Das, N., Dutta, M., Bhattacharjee, S., Chattopadhyay, A., Roy, S. S.  and Bandyopadhyay, D.
 
Oxidative Stress and Free Radical Biology Laboratory, Department of Physiology, University of Calcutta, University College of Science and Technology, 92, A / pc Road, Kolkata 700 009
 E. mail: debasish63@gmail.com
 
 
Abstract: Reactive oxygen species (ROS) are generally produced by the redox imbalance within a tissue, when its metabolic key runs in a frantic reduction mode. These ROS take the major initiative to cause tissue damage in any disease. So removal of ROS may be a major initiative to alleviate tissue damage in various diseased conditions. Our present study was aimed at investigating the protective effect of Terminalia arjuna (TA) aqueous bark extract against ROS induced liver tissue damage using Cu-ascorbate system as an in vitro model of oxidative stress. Incubation of goat liver homogenate with Cu-ascorbate showed an increased level of lipid peroxidation, protein carbonyl content, DNA smearing in agarose gel indicating direct tissue damage. The sharp increase in the activity of pro-oxidant enzymes like xanthine oxidase (XO) is also an indication of tissue damage as this enzyme is involved in formation of uric acid and O2-. The increase in the level of O2- is also evident from increased Cu-Zn SOD activity in cytosol and Mn-SOD activity in mitochondrial fraction. Catalase activity of hepatic tissue is concomitantly decreased, which is an indication of excess H2O2 accumulation in liver tissue. The TA extract dose-dependently prevented the changes of oxidative stress biomarkers, pro-oxidant and antioxidant enzymes in liver tissue when co-incubated with the above mentioned system. These results suggest that TA aqueous bark extract possesses a strong protective ability against oxidative stress in liver tissue, in vitro.
 
Key words: Terminalia arjuna,  Cu-ascorbate, , Oxidative stress, Antioxidant
 
 24. Journal of Cell and Tissue Research 13(2): 3739-3744 (2013)
 
Molecular characterization of micropropagted off type banana (Musa paradisiaca) using DNA markers (ISSR & RAPD)
 
Patel, S. and Anil Kumar
 
Departement of Biotechnology, Govt. V.Y.T.P. G Autonomous College, Durg 491001..
E. mail:aimum_aishley@yahoo.co.in , Cell 09827491253
                    
Abstract : Molecular characterization of the off type of Grand naine variety of micropropagated banana was carried out with the aim to morphologically characterize the off type banana plants under in vitro sub-cultures and hardening stages and their genotyping using ISSR and RAPD markers. In this context six ISSR and three RAPD markers were used which yield 85 and 27 amplicons with an average of 14.16 and 9 bands per marker respectively. The molecular analysis of these off types showed that the variations in these plants are due to the epigenetic factors which do not bring about any change in the genomic sequences.
 
Key words: Musa paradisiaca; Somaclonal Variation; Off types; ISSR; RAPD
 
25. Journal of Cell and Tissue Research 13(2): 3745-3748 (2013)
 
Evaluation of anti-oxidant and hepatoprotective activity of Couroupita guianensis leaves
 
Elumalai, A., Bargavi, K., Krishna, S. and Chinna Eswaraiah, M.
 
Department of Pharmacognosy, Anurag Pharmacy College, Ananthagiri (V), Kodad (M),
Nalgonda 508 206 (A.P.). E. mail: malairx@gmail.com
 
Abstract: The present study was conducted to evaluate the hepatoprotective activity of ethanolic leaf extract of Couroupita guianensis against CCL4 induced liver damage in rats. Anti-oxidant activity was evaluated by DPPH (2, 2-diphenyl-1-picrylhydrazyl) and hydrogen peroxide scavenging. Ascorbic acid was used as the standard antioxidants for comparison. Healthy male Wistar albino rats of either sex (150-200g) were used for the in-vivo investigations. Liver damage was induced by administration of 30% CCl4 suspended in olive oil (1ml/kg body weight). Activities of liver marker enzymes, serum glutamate oxaloacetate transaminase (SGOT), serum glutamic pyruvic transaminase (SGPT), asparate aminotransferase (AST), alkaline phosphatase (ALP), total bilirubin (TB) and total protein (TP) at doses of 150mg/kg and 300 mg/kg showed a significant hepatoprotective effect in comparison with the standard (silymarin). The present study demonstrates that the extract of Couroupita guianensis has hepatoprotective effect against CCl4 induced hepatotoxicity.
 
Key words: Couroupita guianensis, hepatoprotective effect, Silymarin.
 
26. Journal of Cell and Tissue Research 13(2): 3749-3754 (2013)
 
Antioxidant profile of organically and conventionally grown fresh turmeric (Curcuma longa L.): A comparative study
 
Roghelia. V. and Patel, V.H.
 
Laboratory of Foods and Nutrition, P. G. Department of Home Science, Sardar Patel University, Vallabh
Vidyanagar 388120, Gujarat. E. mail: virajshalin@gmail.com, Phone : +91-2692-226877/78
 
Abstract : Turmeric (Curcuma longa) is a an important spice in India from ancient time due to its colouring properties by its key pigment curcumin and medicinal efficacy as anti-atherosclerotic, antidiabetic, antimutagenic and anticancer agents. The effect of cultivation practices on antioxidant capacity of fresh turmeric was evaluated from organically and conventionally grown samples collected from Anand (Gujarat, India). Total phenol, flavonoid, curcumin contents were measured using standard methods. Total antioxidant capacity of fresh turmeric was measured by DPPHRSA, ABTSRSA and FRAP. Results from this study showed that in organically grown turmeric, total phenol, flavonoid and curcumin contents were 708.06 /100g, 84.57mgRutinEq/100g and 41.24 mgRutinEq/100g respectively, which were significant (p<0.01) higher than the same of conventionally grown turmeric. A significant and positive correlation was observed in curcumin with DPPHRSA and FRAP in organically grown turmeric and similar correlation of curcumin with ABTSRSA was observed in conventionally grown turmeric. The study concludes that organically grown turmeric offers higher amount of antioxidant properties and significant differences between two cultivation practices were evident.
 
 Key words : Turmeric (Curcuma longa L.), Antioxidant
 
27. Journal of Cell and Tissue Research 13(2): 3755-3759 (2013)
 
Callus development and indirect shoot regeneration from leaves and stem explants of Stevia  Rebaudiana bertoni
 
Ojha, A. and Choudhary, R.
 
Department of Biotechnology, Meerut Institute of Engg. and Technology, Meerut 250 005, U.P.;
 E. mail: ashishbt26@gmail.com, Cell 09996640894
 
Abstract: A protocol for callus induction and multiplication was developed to produce large no. of calli in short period. Surface sterilized leaf and Stem explants were cultured on Murashige and Skoog (MS) medium with different concentration and different combination of plant hormone like, IAA, kinetin, NAA, 2,4-D, and BAP with alone and combinations. 100% callusing was observed from leaf explants cultured on combination of low 2,4-D and high NAA (0.5 mg/L-1 +1.5 mg/L-1 ) after 12 days while alone 2,4-D only 30 % callusing was observed. 68% callusing was observed in 0.5 mg/L-1 BAP + 0.5 mg/L-1 2, 4-D from stem explants. 14 % callusing was observed in combination of high concentration of auxines and cytokinins (1.5 mg/L-1 + 1.5 mg/L-1 2,4-D) in stem explants were shiny green. The maximum number of shoot (9.1±0.4) was observed on the MS medium with BAP (2.0 mg/L-1). The well Develop shoots used for rooting in vitro and 100% rooting found in 0.5 mg/L-1 IBA. The in vitro developed rooted plantlets were transferred to medium size pots and they were grown in greenhouse for hardening and finally they were planted in the open filed. Around 92% of plants were successfully established in natural field condition. The objective of present study was to develope a simple, rapid, economical, and high frequency regeneration protocol from leaf and stem explants of S. rebaudiana for potential application in large scale propagation.
 
Key words: Stevia rebaudiana, shoot regeneration, explants and auxins
 
28. Journal of  Cell and Tissue Research 13(2): 3761-3764 (2013)
 
Effect of Parsley extract on sublingual gland of naturally aged male mice
 
Khandare, S.N., Pillai, M.M. and Khandare, N. K.
 
Vidnyan Mahavidyalaya, Sangola, Dist. Solapur 413 307 (MS). E. mail: khandaresupriyanil@yahoo.com
 
Abstract: The aging is generally characterized by the declining in ability to respond to stress, increasing homeostatic imbalance and increased risk of diseases. Aging promotes free radical formation in the cell. Free radicals formed due to aging or various reasons are scavenged by   antioxidants. In present study antioxidant rich plant, Petroselinum crispum (mill), was used to assess to remove free radicals generated during aging. Three groups of mice (control, naturally aged and parsley receiving) were used during the experiments. Study was restricted sublingual gland. Histological sections of glands were stained with haematoxylene and eosine. Study shows damage in lingual gland in agd mice as compare to young control. Parsley (Petroselinum crispum), also corrected the histological structure of sublingual gland as well as weight of mice in naturally aged animal.
 
Key words: - Aging, sublingual gland, Parsley
 
29. Journal of Cell and Tissue Research 13(2): 3765-3768 (2013)
 
Callus induction and plantlet regeneration in tomato (Solanum lycopersicum L.)
 
Bahurupe, J.V., Patil, S.C., Pawar, B.D., Chimote V.P. and Kale, A.A.
 
State Level Biotechnology Centre, Mahatma Phule Krishi Vidyapeeth, Rahuri-413 722, Maharashtra.
E. mail: jyotibahurupe2012@gmail.com
 
Abstract: In vitro plant regeneration protocol was developed using shoot tip and hypocotyl explants in tomato variety ‘Dhanashree’. Earliest callus initiation was observed within 6.73 days in shoot tip explants and in 7.53 days in hypocotyl explants. Shoot tip explants showed 100% callus induction when cultured on MS medium supplemented with 2.0 mg/l benzylaminopurine (BAP) and 0.2 mg/l indole acetic acid (IAA). The highest frequency of shoot regeneration (78.40 and 74.93 %) and maximum number of shoots per explant (4.90 and 4.33) was obtained from the hypocotyl and shoot tip explants, respectively. Developed shoots were subjected to rooting on MS medium devoid of growth regulators and the rooted plants were hardened in the greenhouse.
 
Key words: Solanum lycopersicum;Callus induction
 
30. Journal of Cell and Tissue Research 13(2): 3769-3774 (2013)
 
Isolation and characterization of potential probiotic lactic acid bacteria from human infant fecal samples
 
Pandya, N., Patel, V.H., Subhash, R. and Elias, J.
 
P. G. Department of Home Science, Sardar Patel University, Vallabh Vidyanagar, Anand, Gujarat. E. mail: jishaelias@yahoo.co.in
 
 
Abstract: Lactose intolerance, diarrhea, colon cancer and various gastrointestinal disorders are most commonly found in the developing countries. Control of these could be achieved by potentially probiotic lactic acid bacteria. Primary aim of this study was to isolate lactic acid bacteria from the feces of 25 days old breast fed infants and analyzing their probiotic properties such as acid resistance, bile resistance, antimicrobial activity, antibiotic resistance, carbohydrate utilization and milk fermentation. Approximately twenty isolates from infant fecal sample were screened for their antimicrobial activity. Out of these, three strains were able to meet the basic selection criteria of probiotics. These three isolates were tolerant to pH 2 (45 min.) and could grow in 4% oxgall (45 min.). They showed good antimicrobial activity against S. typhi, S. aureus, E. coli. The isolates were also found to be able to ferment a variety of sugars and were also resistant against a large number of antibiotics (penicillin, ampicillin, amphotericin b, amoxicillin, clostrimazole and ciprofloxacin). These three isolates showed efficient curdling ability in 4 hours. Out of three strains two were identified to be Enterococcus faceium by 16 s r DNA-cataloguing. Enterococcus faceium isolated from infant fecal sample possess potentially probiotic properties and could be effective against enteropathogens.
 
Key words: Probiotics, Lactic acid bacteria
 
31. Journal of Cell and Tissue Research 13(2): 3775-3778 (2013)
 
Antiovulatory and antiimplantation effect of ethanolic extract of root of Achyranthes aspera Linn. in the female rat
 
Bodare, R.D., Birje, K.N., Bansode, D.L., Deshmukh, A.J., Inamdar, I.I., Mane, A.B. and More, R.B.
 
Post Graduate Department of Zoology, S. G. M. College, Karad 415 124 (M. S.).
E. mail: bodare_62@rediffmail.com
 
Abstract: The antifertility effect of Achyranthes aspera was studied in the female Wister rats. Antiovulatory effect was assessed by observing oestrous cycle and antiimplantation effect was studied by observing the implantation sites in the uterus. The ethanolic root extract of Achyranthes aspera arrested the nomal oestrous cycle at dioestrous phase. The oestrous cycle of treated rats resumes to normal after withdrawal of treatment. The control showed normal oestrous cycle. There was decrease in the number of the implantation sites in the treated rats as compared to the controls. The results suggest that ethanolic extract of root of Achyranthes aspera has antiovulatory and antiimplantation properties which were found to be reversible.
 
Key words: Achyranthes aspera, Antiovulatory, Antiimplantation
 
 
 
 
 
 
 
 

 
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