Botany - Mphil Thesis

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Selection for Salt Tolerance in Petunia Grandiflora
(Central University of Punjab, 2014) Najar, Raoof Ahmad; Saini, R.G.
Petunia grandiflora a native of South America is a popular and high value ornamental plant in the world. Soil salinity is the major abiotic stress in semi-arid Malwa region of Punjab, adversely affecting its productivity, survival and quality. In order to develop salt resistant varieties of P. grandiflora, an attempt was made to select salt tolerant seedlings from varieties Violet Blue, Giant California and Nana compecta by using ex-vitro and in-vitro methods. Seedlings were subjected to salt treatments of 100 mM, 200 mM, 300 mM, 400 mM, 500 mM and 600 mM for durations of 4 hr, 6 hr, 12 hr and 24 hours. Salt treatment reduced shoot length, leaf number and survival percentages and delayed days to 50% flowering. Variety Nana Compecta was identified as tolerant to salt and variety Giant California was most sensitive to soil salinity. In in-vitro method of gradual increase in NaCl concentration (0, 25, 50, 75 and 150 mM) from low to high level was found to be a better approach for selecting salt tolerant calli as compared to direct method in which direct transfer of calli to high salt concentrations (50, 100, 150, 200, 250 and 300 mM) was found to be detrimental to callus survival and growth. Treatment of calli upto 50 mM salt concentrations was good for shoot regeneration in all the three varieties. Protocol was standardized for callus induction, direct shoot induction from leaves, shoot regeneration from callus and root induction from shoots. For callus induction, 2 mg/l 2, 4-D and 0.5 mg/l kinetin was optimum for variety Violet Blue and 1 mg/l both BAP and NAA was optimum for varieties Giant California and Nana Compecta. For direct shoot induction from leaves 2 mg/l BAP and 0.1 mg/l NAA were good for all the three varieties. For shoot regeneration from calli, 2 mg/l BAP in combination with 0.1 IAA for variety Violet Blue and 1 mg/l both BAP and NAA for varieties Giant California and Nana Compecta were ideal. For root induction, 0.5 mg/l NAA and 0.1 mg/l IBA were ideal for all the three varieties. Seeds from plants grown ex-vitro showing tolerance to salt were harvested and stored for further testing.
Development and characterization of genomic microsatellite markers in Melia azedarach
(Central University of Punjab, 2014) Thakur, Sapna; Bhardwaj, Pankaj
Melia azedarach is ecologically imperative species known for its innumerable biological benefits such as antiviral, anthelminthic, antibacterial, etc. In this study, we developed 43 genomic microsatellite markers from (AG)n enriched library and subsequently employed 23 of them for genetic diversity and population structure analysis of Melia azedarach growing in Indian Thar desert. Fourteen populations encompassing 95 genotypes were selected for analysis and we found a moderate level of diversity (Na = 3.211, Ho = 0.558, He = 0.549, P = 94.41%) in them. Gene diversity (h) among population pairs varied from 0.566 to 0.714 with very low overall genetic differentiation (F = 0.021). The highest value of ΔK estimated using STRUCTURE indicated 2 subpopulations (K=2) and admixed cluster occupied maximum area (75.79%) under Bar plot. Genetic distance based UPGMA dendrogram also identified 2 major clusters among 14 Melia azedarach populations. UNJ tree based on genetic dissimilarity clustered genotypes from different population together. No significant correlation between geographical and genetic distance was found in present study (Rxy = 0.261, P = 0.18). Allele frequency distribution under “mode-shift” indicator was normal L-shaped, suggesting populations under study are not experiencing any recent bottleneck. This study laid the foundation for more precise inference about the biogeography and management of M. azedarach in the Indian Thar Desert
To study the effects of insulin and metformin on pc-3 cell line
(Central University of Punjab, 2016) Singh, Pushpendra; Bast, Felix
Dietary habits, genetic factors, hormonal factors and environmental factors are the independent risk factors for prostate cancer as well as diabetes. Androgen is the primary growth factor for the prostate cancer initiation and progression, however, non androgen peptide growth factor like insulin and insulin growth factor also involved in the prostate cancer as well as diabetes. Insulin and insulin growth factor are peptide that regulates metabolism, growth, cellular proliferation and apoptosis. The anti-diabetic drug metformin is rapidly emerging as a potential anti-cancer agent that improves insulin homeostasis and decreased growth and cellular proliferation of the prostate cancer cell line. Thus it is necessary to understand the growth promoting role of insulin on prostate cancer cell line and the possible influences of metformin on the proliferation of prostate cancer cell line in the presence and absence of insulin has been studied.
Development and chacterization of genomic microsatellite markers in rhdodendron arboreum
(Central University of Punjab, 2014) Choudhary, Shruti; Bhardwaj, Pankaj
Rhododendron arboreum, a key species inhabiting Indian Himalayas, has ecological and economic importance. Global climate change and anthropogenic activities can pose a threat to the biodiversity richness. Population genetics characteristics are the fundamentals of conservation and management practices. However, no genetic background for the species is presently available. To provide an insight into the population structure of the species, an effective marker resource was developed. A genomic library enriched for microsatellites was constructed using biotin- streptavidin hybridization technique, followed by sequencing of positive clones. With 54% enrichment rate, 41 primers were designed from SSR clones, consisting of perfect or interrupted repeats of AG/CT, AC/GT motifs. 38 loci showed successful amplification on genotypes of three populations with number of alleles ranging from 2 to 14, observed and expected heterozygosity of 0.167 to 0.933 and 0.422 to 0.917, respectively and average polymorphic information content of 0.104 to 0.811. A high gene flow (Nm= 5.436) demonstrated high genetic diversity within the populations. 19 loci displayed significant deviations from Hardy Weinberg equilibrium. Overall no linkage disequilibrium and bottleneck was detected among sampled populations. DARwin and STRUCTURE analysis grouped the populations into two original clusters and admixed genetic stock irrespective of their geographical locations. These novel loci will support further genotyping studies in R. arboreum.
Selection for salt tolrance in petunia grandiflora
(Central University of Punjab, 2014) Najar, Raoof Ahmad; Saini,R.G
Petunia grandiflora a native of South America is a popular and high value ornamental plant in the world. Soil salinity is the major abiotic stress in semi-arid Malwa region of Punjab, adversely affecting its productivity, survival and quality. In order to develop salt resistant varieties of P. grandiflora, an attempt was made to select salt tolerant seedlings from varieties Violet Blue, Giant California and Nana compecta by using ex-vitro and in-vitro methods. Seedlings were subjected to salt treatments of 100 mM, 200 mM, 300 mM, 400 mM, 500 mM and 600 mM for durations of 4 hr, 6 hr, 12 hr and 24 hours. Salt treatment reduced shoot length, leaf number and survival percentages and delayed days to 50% flowering. Variety Nana Compecta was identified as tolerant to salt and variety Giant California was most sensitive to soil salinity. In in-vitro method of gradual increase in NaCl concentration (0, 25, 50, 75 and 150 mM) from low to high level was found to be a better approach for selecting salt tolerant calli as compared to direct method in which direct transfer of calli to high salt concentrations (50, 100, 150, 200, 250 and 300 mM) was found to be detrimental to callus survival and growth. Treatment of calli upto 50 mM salt concentrations was good for shoot regeneration in all the three varieties. Protocol was standardized for callus induction, direct shoot induction from leaves, shoot regeneration from callus and root induction from shoots. For callus induction, 2 mg/l 2, 4-D and 0.5 mg/l kinetin was optimum for variety Violet Blue and 1 mg/l both BAP and NAA was optimum for varieties Giant California and Nana Compecta. For direct shoot induction from leaves 2 mg/l BAP and 0.1 mg/l NAA were good for all the three varieties. For shoot regeneration from calli, 2 mg/l BAP in combination with 0.1 IAA for variety Violet Blue and 1 mg/l both BAP and NAA for varieties Giant California and Nana Compecta were ideal. For root induction, 0.5 mg/l NAA and 0.1 mg/l IBA were ideal for all the three varieties. Seeds from plants grown ex-vitro showing tolerance to salt were harvested and stored for further testing.
Sequence-based phygeography and conservation of seaweeds from indian subcontinents
(Central University of Punjab, 2014) Rani, Pooja; Bast, Felix
Phylogeography is the study of historical events which are responsible for evolution and current distribution of a species in different geographical area. However very less record is available about marine macroalgae of Indian subcontinent. This study investigated the DNA barcoding and phylogeographical distribution of marine algae from the Indian subcontinent. Different algae samples collected from various coasts of Indian subcontinent are amplified using ITS, COX and rbcL primers. In our results, we found the occurrence of green algae like Ulva reticulata, Ulva intestinalis, Ulva fasciata, Ulva proliifera, Ulva ohnoi and one sample with Caulerpa scalpelliformis ; Red algae, Gracilaria foliifera, Gracilaria domingensis, Gracilaria corticata, Grateloupia Sp., Ceramium Sp., Centroceras clavulatum, Erythrocladia Sp., Erythrocladia irregularis, Acanthophora Sp., Dilsea socialis, Hypnea stelullifera, Sirodotia tenuissima and Dichotomaria Sp.; Brown algae, Sargassum zhangii, Sargassum megalocystum, Sargassum aquifolium and Turbinaria ornata in Indian subcontinent. Gracilaria domingenesis, Dilsea socialis, Sargassum megalosystum were first time reported in India. On the basis of molecular studies, we found that Ceramium Sp. Nov., Erythrocladia Sp. Nov., Acanthophora Sp Nov., Grateloupia Sp. Nov. and Dichotomaria Sp. Nov. were identified as new species. Erythrocladia irregularis was identified as endophytic algae inside green algae Cladophora glomerata. Phylogenetic tree was generated to analyse the evolutionary distance between different samples. Morphological and microscopic studies were performed for each sample. This study further helps in identification and documentation of new species and cryptic species. All samples were pressed for herbarium voucher.
DNA Barcoding and phylogography of brown seaweeds of coasts of indian subcontinent
(Central University of Punjab, 2013) Bhushan, Satej; Bast, Felix
Algae are one of the diverse groups of ubiquitous autotrophs. Their use as food was more or less initially confined to few East Asian and South American countries like China, Japan, Korea, Chile, Argentina and Brazil, but with the increased demand and limited supply for food combined with the medicinal properties of the marine macroalgae, they started getting the attention of policy makers and researchers alike all around the world. Brown seaweeds (Phaeophyceae) are mostly marine and characterized by presence of pigment fucoxanthin which gives them its coloured appearance. The present work aims to provide detailed molecular analysis of the brown seaweeds found in Indian coastal regions to study and characterize it taxonomically which has not been done till now in Indian context. Out of all the samples processed, one invasive species was detected, Sargassum zhangii, which is the first report of this algal species outside Chinese waters. The conspecificity was confirmed by a multi-faceted approach, including comparative morphology, microscopy, genetic distance analysis and computational phylogenetics using Maximum Likelihood and Bayesian Inference methods.
Development and charaterization of UGMS markers for genetic diversity analysis in Rhododendron arboreum
(Central University of Punjab, 2013) Sharma, Gagan; Bhardwaj, Pankaj
Rhododendron arboreum is an ecologically important species growing in Himalayan regions. It is widely popular due to its medicinal potential, edible, sacred & economic value, however very little is known about its genetic diversity. The development and characterization of UGMS markers will provide a major impact on genetic diversity analysis. Total 792 (615 singletons and 177 contigs) unigenes were predicted from 1,241 publicly available EST database from NCBI (http://www.ncbi.nlm.nih.gov/) site in R.catawbiense by clustering of 2 - 33 random EST sequences. From 50 SSR contained Unigenes, 36 UGMS primers were designed for the cross amplification approach and six pairs of microsatellite primers were produced clear PCR amplification. The cross related species/genera transferability rate was 16.66% from R.catawbiense to R.arboreum. The polymorphism information content (PIC) ranged from 0.2756 to 0.9212 with an average of 0.5765.The average observed heterozygosity (Ho) was 0.8666 i.e. greater than average expected heterozygosity (He) was 0.6792. The genetic diversity was found with an average of 0.6222 within population of R.arboreum and genetic diversity found with an average of 0.3436 among the population.This study revealed the insight of abundance & distribution of microsatellite in the expressed region of the Rhododendron arboreum genome.
Preconditioning of chickpea seedlings for terminal heat stress: understanding associated mechanism and HSP's expression
(Central University of Punjab, 2014) Yadav, Renu; Kumar, Sanjeev
Chickpea (Cicer arietinium L.) is the second most important pulse crop grown worldwide. Changes in the cropping system, competition from other cash crops (wheat) and global warming are pushing chickpea to relatively warmer growing environment. In northern part of country chickpea come across with terminal high temperature stress during reproductive stage which lead to reduced grain yield. Therefore to prevent the plant from incoming heat stress, 11 day chickpea seedling were preconditioned with mild drought stress, then put on recovery for six days and then recovered seedlings were exposed to lethal stress (where temperature was increased step wise from 30?C to 36?C). This study revealed that % EL, Lipid peroxidation increased with the increase in temperature while percent TTC reduction and total protein content decreased with the increase in temperature. Antioxidative enzymes provide the major defence against the ROS generated during the abiotic stress, and it was found that activity of SOD, CAT and APX enzyme increased proportionately with the rising temperature. HSP's act as molecular chaperons and are over expressed at both mRNA and protein level in preconditioned seedlings exposed to high temperature stress as compared to non- preconditioned ones. To conclude the whole study, results obtained clearly reveal that preconditioning with drought stress has the ability to improve tolerance above ambient temperature (27?C 7?C), thereafter preconditioning did not have any influence in terms of the improvement in membrane damage and level of antioxidants. Higher expression of sHSP's is corroborated with the low expression of antioxidants.
Preconditiong chickpea for chilling stress: Understanding associated mechanisms and role of FADs in membrane protection
(Central University of Punjab, 2014) Saini, Rashmi; Kumar, Sanjeev
Chickpea has evolved as a spring crop in warmer climate of West Asia. Therefore, it is prone to chilling injury as in North India, chickpea is cultivated in winter season. Preconditioning the plants with mild drought stress might triggers the signaling pathways that will prepare the plants to combat lethal cold stress. In this study, one chilling tolerant genotype PDG3 and one chilling sensitive genotype GPF2 of chickpea were preconditioned with mild drought stress and then exposed to lethal cold stress at 40C. To see the effects of preconditioning various cell responses were monitored by measuring ELI, MDA content, LOX activity, antioxidative enzymes SOD, CAT, APX, GPOX and GR, and mRNA expression of fatty acid desaturase (FAD-2) and small heat shock proteins (sHsp18.5 & sHsp22.7) at different phases. Both the genotypes behave differently to preconditioning. This study shows that preconditioning increases the tolerance capacity of chilling-sensitive genotype towards chilling stress. Whereas, preconditioning brings about adaptable changes in chilling tolerant plant PDG3 so, they can cope up with chilling stress. To conclude whole study, preconditioning with mild drought stress has ability to improve the tolerance against cold stress in chickpea.
Oxidative stress and innate immune response in A549 lung carcinoma cells
(Central University of Punjab, 2014) Upadhyay, Shishir; Dhiman, Manisha
Cancer immunology is the study of cross-talk between the immune response and cancer cells. The immune response, including the recognition of cancer-specific antigens is of particular interest as knowledge gained drives the development of new vaccines and antibody therapies. The molecular mechanisms which are disturbed in the susceptible patients who proceed to develop cancer are very complicated and still largely unknown. It proposed that apart from the reported genetic modifications on tumor cells there are modifications due to oxidative stress (resulting in the formation of chemical adducts, e.g. 3-hydroxynonenal, 3- nitrotyrosine, carbonyl etc.) at the vicinity of tumor where the immune cells infiltrate. The central hypothesis of the present work is that respiratory burst which is host?s mechanism to kill the foreign particles (tumor cells) is used as defence mechanism by the tumor cells by forming neoantigen which in turn make them undetectable and can further help them to escape the host immune surveillance. The lung carcinoma A549 cells were treated with100μM H2O2 and using 1-D gel electrophoresis the oxidized tumor proteins in normal and treated cells were visualized. To confirm the oxidized modifications at membrane levels and at proteins levels the lipid peroxidation and protein carbonyls were detected respectively. It was observed that the oxidative stress induces the lipid peroxidation and protein carbonyls in tumor cells. To determine if neo (oxidized) tumor antigens elicit any alteration in immune responses where they show compromised phagocytosis, thus resulting in a failure to elicit effector immune functions were analyzed via phagocytosis and respiratory burst using spectrophotometry and microscopic techniques. The present study has identified a novel mechanism(s) of carcinogenesis initiation and which can further provide directions for the development of adjunct therapies to control cancer in its initial stages and at the same time it advocates for new ventures to increase the efficacy of the chemotherapeutic drugs.
Isolation and characterization of heavy metal resistant bacteria from soil
(Central University of Punjab, 2014) Mehra, Richa; Saini,R.G
The exploitation of microbes to remove heavy metals from contaminated sites and industrial effluents is an extensive area of research. In this study, isolation of heavy metal resistant bacteria was done from potentially heavy metal contaminated agricultural soil of Bathinda region. The ICAP-AES analysis of soil revealed toxic concentrations of lead and nickel whereas most other heavy metals were also present in considerable amounts. The organisms showing maximum resistance to eight heavy metals tested (Pb, Cu, Ni, Cr, Co, Cd, Fe and As) were screened. The 29 potent metal resistant organisms were subjected to biochemical characterization and 16S rDNA sequencing, and tentatively identified as strains of Bacillus and Streptomyces spp. All the strains showed multiple heavy metal resistance with varied degrees of resistance. A strain of B. megaterium FE1 exhibited maximum resistance to all the heavy metals- Pb (2.5 mg/ml), Cu (1.5 mg/ml), Ni (1.0 mg/ml), Cr (3.0 mg/ml), Co (0.5 mg/ml), Cd (2.5 mg/ml) and As (2.5 mg/ml). The incidence of such extent of heavy metal resistance in native bacteria suggest that agricultural soil is undergoing stress which needs more intensive studies.
Diversity of green and red macroalgee distributed in indian west-coast using morphometry and DNA barcoding
(Central University of Punjab, 2013) Aijaz Ahmad John; Bast, Felix
Modern algal systematists exploit genetic tools for molecular assisted alpha taxonomy and DNA barcoding is one such molecular tool that relies on the use of a standardized DNA region as a tag for rapid and accurate species identification. In this study Nuclear Internal Transcribed Spacer (ITS) was used as a marker for identification and phylogenetic analysis of marine green and red macroalgae in Indian subcontinent. Using Bayesian Inference for phylogenetic reconstruction with T3P model of molecular evolution and gamma distribution (T3P+G) using ITS dataset revealed that the bloom forming Ulva intestinalis found in west coast of Indian subcontinent showed strong endemism, supporting the view that that the genus Ulva encompasses a number of endemic cryptic species in addition to cosmopolitan species. Although there were two morphotypes present in Indian isolates, they constituted a single clade with robust Bayesian Posterior Probability support, confirming conspecificity of these morphotypes. Our results also indicate latitudinal gradients in the distribution of tubular Ulva, with a clade encompassing all nontropical isolates. Higher genetic heterogeneity of tropical isolates as evidenced by highest within-group T3P (Tamura-3-Parameter) distances comparing with that of non-tropical isolates is suggestive of tropics being the geographic origin of these species. While U. compressa and U. intestinalis were monophyletic within nontropical superclade, these morphotypes were polyphyletic within the tropical clade. Due to the polyphyly of currently accepted morphospecies concept and formation of distinct phylogenetic clade among Indian isolates forces us to propose a new bloom forming species of Ulva paschima. Further molecular assessment of invasive Carrageenophyte Kappaphycus alvarezii using ITS-1 region showed affinity to phylogenetic clade of mixed geographical origin confirming that the species was introduced in the subcontinent by human intervention. Surprising result of our study was an endophytic green algae Ulvella leptochaete that was found growing inside Caldophora glomerata, a first report of its kind from India.
Effect of amyloid beta (25-35) peptide on mitochondrial respiratiory function in neuroral cells over expressing ape1
(Central University of Punjab, 2014) Kaur, Navrattam; Mantha, Anil K.
Alzheimer's disease (AD) is an important public health problem which affects millions of people worldwide. The major pathological hallmarks associated with AD are the accumulation of amyloid beta (A?) in senile plaques and neurofibrillary tangles (NFTs) made up of hyperphosphorylated tau proteins. Accumulating evidences point towards the role of oxidative stress and mitochondrial dysfunction in the pathogenesis of AD. Aging is considered as one of the greatest risk factor for AD. In order to maintain genome integrity, base excision repair (BER) pathway is the predominant pathway for repairing oxidized base lesions in neuronal cells. APE1 is the central enzyme of the BER-pathway, having both repair and redox activities and shown to enhance neuronal survival after oxidative stress. In my study, effect of A?(25-35) on mitochondrial ROS/RNS levels and activities of respiratory complexes (I, III, & IV) in neuronal cells was studied with and without ectopic APE1 expression and the neuro- modulatory role of Ginkgolide B (from leaves of G. biloba) was evaluated. It was seen that A?(25-35) increases the ROS/RNS levels in these cells which was decreased when pre-treated with Ginkgolide B (G.B) before treating with A?(25-35). APE1 levels were found to be decreased on treating with A?(25-35) and were increased on pre- treatment with G.B and subsequent treatment with A?(25-35). These results indicate that ectopic APE1 expression in the mitochondria of the neuronal cells might overcome the oxidative damage caused by A?(25-35). Also, phytochemical G.B has shown to modulate the mitochondrial complex activity upon A?(25-35)-induced oxidative stress and modulate the ROS/RNS levels in the presence of APE1. Further studies are needed to understand the mechanism of action of APE1 in relation to the above results, which will be carried out during my Ph.D. work.
Phytogenetic analysis of bacterial diversity of heavy metal affected
(Central University of Punjab, 2012) Kumar, Rajeev; Kumar, Sanjeev
The soil of Bathinda region is affected by heavy metals due to anthropogenic activity, industrial effluent and fly ash. Three soil samples in triplicate were collected from three different places of Bathinda region, abandoned soil, agricultural soil, thermal power plant fly ash soil. Based on 16S rRNA gene cloning and sequence analysis, phylogentic analysis of bacterial diversity was performed. 366 clones were picked and through colony PCR 128 positive clones were chosen. Out of 128 clones, 126 clones were sequenced in three fragments and all three fragments aligned using SeqMan Lasergene ver.10.0 (DNA STAR) software. BLAST analysis of sequenced and aligned samples was carried out at NCBI to find out the homology with different groups of bacteria. All 126 clones aligned for phylogenetic analysis by MEGA 5 (Neighbor Joining method). 32 genus belong to 8 phyla were observed in all three samples. Most dominant phylum is Proteobacteria followed by Bacteriodates. Pseudomonas spp. is dominant in agricultural and thermal power plant soil. Abandoned soil showed maximum diversity followed by thermal power plant soil. In agricultural soil minimum diversity was observed. So with this investigation it can be concluded that lands without anthropogenic activities are rich in bacterial diversity as compared to more anthropogenic active areas.
Cytotoxic activity of saragassum wightii on pc-3 cancer cell-line
(Central University of Punjab, 2012) Jahid, Mohd.; Bast, Felix
Finding novel antitumor compounds with low side effects could be an interesting proposal. Antioxidant and anticancer potentials of seaweed extracts can possibly be explored for developing the new anticancer drugs. Polyphenols are extensively distributed in seaweeds and these are reported to be free radical scavengers. This study was designed to check the cytotoxic potential of the methanolic and hydromethanolic extracts from the brown algae S. wightii. The total phenolic content in the hydromethanolic extract and the methanolic extract of S. wightii was determined by the Folin-Ciocalteu method. The total phenolic content in methanolic extract was found to be greater than hydromethanolic extract. The percentage inhibition or scavenging activity of both the extracts was calculated by using the DPPH assay and was more for the methanolic extract. PC-3 cancer cell-line was used as an experimental model. The methanolic crude extract is significantly cytotoxic against the PC-3 cancer cells. On the other hand the hydromethanolic crude extract was not found to be the significantly cytotoxic against the PC-3 cancer cells.
To Study the Dose and Time Dependent Effect of Human Insulin and Metformin on the Growth of Breast Cancer Cells
(Central University of Punjab, 2012) Cholia, Ravi Prakash; Thakur, Sanjeev
Cancer and diabetes, both are leading causes of mortality globally. Both the diseases are multifactorial and share number of common risk factors. Hyperglycemia and hyperinsulinemia which are the characteristic features of diabetes influences the growth rate and proliferation of tumor cells directly or indirectly. Type 2 diabetes shows stronger association with various cancers. Breast cancer is one of the malignancy affecting females worldwide. This study demonstrates that glucose not only acts as energy source in tumor cell but also acts as mitogen. Insulin not only regulates the blood glucose level but also induces growth and proliferation in MCF 7 and MDA MB 231 breast cancer cell lines independently and in combination with glucose. Metformin inhibit proliferation of MCF 7 and MDA MB 231 breast cancer cell lines independently and also in presence of glucose and insulin, but shows more inhibitory effect in presence of insulin as compare to glucose. Recently discovered insulin receptor antagonist S961 did not showed any significant response in breast cancer cell lines MCF 7 and MDA MB 231. The ineffectiveness is probably due to blocking effect of higher insulin dose. So with this investigation it can be concluded that metabolic alteration leads to proliferation of breast cancer cell lines.
To study the effect of insulin on advanced androgen-indpendent prostate cancer (pc-3) cells
(Central University of Punjab, 2012) Kumar, Abhimanyu; Kumar, Sanjeev
Prostate cancer is one of the major causes of mortality in males over the age of fifty all over the world. Many factors including genetics and diet have been associated with the development of prostate cancer. Hyperinsulinemia has been found to be associated with higher risk of Prostate cancer. Diabetes type-2 is accompanied with hyperinsulinemic state. Both cancer and diabetes are metabolic disorders and often diabetes is correlated with cancer. This study reveals that insulin acts as a mitogen hence increases proliferation in PC-3 cells. Reactive oxygen species are by product of cellular metabolism. Insulin treatment increases cellular metabolism due to which ROS level also increases at higher insulin doses. ROS is necessary for many cells signalling process, abnormal increase in ROS level can cause mutational DNA damage and affects protein folding. Antioxidants and free radical balance is critical for normal cellular functioning. Superoxide dismutase is an important antioxidant enzyme, which keeps ROS level low by dismutation of superoxide anion into hydrogen peroxide. This is further metabolised by catalase. In our study we have found that at lower insulin doses SOD level increases but at higher insulin doses SOD expression decreases significantly. This may be the possible reason of ROS increase. Matrix metalloproteinase's expression is modulated by insulin, which can lead to increase in malignancy. All factors stated above indicate that hyperinsulinemia can lead to tumor progression.
Genetic diversity and polpulation analysis of melia azaderach L. by Rapd markers in bathinda and adjoing regions
(Central University of Punjab, 2013) Navgeet; Bhardwaj, Pankaj
Melia azedarach L. is an ecologically imperative species growing in Thar Desert. Biological effects such as antiviral, anthelminthic, antibacterial etc. makes it important, yet a little is known about its genetic diversity and structure. In this study, 30 RAPD primers were employed for DNA profiling of 47 individuals representing 6 populations from different geographic locales. A total of 87 bands were scored with an average of 2.9 bands per primer. The PIC ranged from 0.1195 to 0.4998 with an average of 0.4160. Nei's genetic diversity (h) and Shannon's information index (I) ranged from 0 to 0.5 and 0 to 0.6931 with an average Nei's genetic diversity of 0.2422. The gene flow (Nm) 1.4381 and the genetic diversity of 0.2381 at species level demonstrated overall high level of genetic diversity. Cladistics analysis using DARwin and Bayesian cluster analysis using STRUCTURE placed 47 individuals into two main clusters or original genetic stocks (K=2) which showed little or no association with the geographic origin. The lack of clear assignment of individuals to geographic regions of sampling and consideration of different populations in the same genetic cluster suggests a recent common evolutionary history.
Population analysis of media azesarach L. by RAPD markers in the region of rajasthan
(Central University of Punjab, 2013) Ahmad, Kamal; Bhardwaj, Pankaj
Melia azedarach L is ecologically imperative species growing in Thar Desert. Biological effects such as antiviral, anthelminthic, antibacterial, etc. makes it important, yet a little is known about its genetic diversity and structure. In this study, we employed 30 RAPD primers for DNA profiling of 91 individuals representing 8 geographically isolated populations. A total of 98 bands were scored with an average of 3.2 bands per primer. The PIC ranged from 0.0681 to 0.5351 with an average of 0.4103. Nei's genetic diversity (h) and Shannon's information index (I) ranged from 0.1237 to 0.2375 and 0.1845 to 0.3500 respectively with average Nei'sgenetic diversity of 0.2570. The gene flow (Nm) 0.9295 and the genetic diversity of 0.3498 at species level demonstrated overall high level of genetic diversity. Cladistics analysis using DARwin and Bayesian cluster analysis using STRUCTURE placed 44 individuals into two main clusters or original genetic stocks (K = 2) which show little or no association with the geographic origin. The lack of clear assignment of individuals to geographical regions of sampling and consideration of different populations in the same genetic cluster suggests a recent common evolutionary history. M. azedarach is a good reforestation tree because of its fast growth and drought hardy nature. This study adds a foundation for more precise inference about the biogeography and management in the reforestation projects in the Thar Desert.

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