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Item Monostroma: the Jeweled Seaweed for Future Cultivation methods, Ecophysiology, Phylogeography and Molecular Systematics(Lambert Academic Publishing, 2011) Bast, FelixThis thesis aims at providing a comprehensive picture of the reproductive physiology, growth, phylogeography, and phylogenetics of M. latissimum-nitidum complex in Southern Japan. A review on agronomy and utilization of seaweeds is provided as Chapter 2 to present a comprehensive overview of the seaweed biology and applications. Seasonality in the growth and occurrence of Monostroma sp. at three environmentally distinct habitats along Tosa Bay, Kochi Prefecture, Japan is explored and results of correlation analyses between environmental conditions and thallus size are presented in Chapter 3. Also investigated in the same chapter is the homology of nuclear encoded rDNA Internal Transcribed Spacer (ITS) sequences between naturally occurring and commercially cultivated populations. Findings of the culture studies sought to identify the species naturally occurring at the study sites are also summarized in Chapter 2. In the course of research, I observed that thalli of the naturally occurring populations of M. latissimum, changes its color during maturation, as reported elsewhere in the literature. Further to that observation, a thorough cytological investigation on the gametangial ontogeny of naturally occurring M. latissimum is presented in Chapter 4 and possible taxonomic implications of this finding are discussed. Once the thalli of M. latissimum release gametes, which mode of syngamy do they have? Is sex of the progenies environmentally determined? These are some of the questions being investigated in Chapter 5. Reported in the Chapter 6 is a serendipitous discovery of an asexually reproducing ecotype of M. latissimum in the marginal populations at low-saline habitat. Findings of the culture studies to complete its life cycle are presented. Molecular studies to investigate homogeneity of nrDNA ITS sequences between the two ecot ypes (i.e., sexual vs asexual) are also investigated in the same chapter. Results of phylogenetic analyses of the newly generated ITS sequence of M. latissimum with that of the related monostromatic green algal taxa retrieved from GenBank are also presented in Chapter 6 to understand relative taxonomical position of this species in the class Ulvophyceae. Chapter 7 is an investigation on the morphologic and genetic homogeneity of natural and cultivated populations of the two closely related species M. latissimum and M. nitidum along the Southern Japanese coast where the warm-water Kuroshio Current influences throughout the year. Combined phylogeographical analysis of nuclear encoded first internal transcribed spacer (ITS1) sequences and rDNA 18s gene are presented in addition to the comparison of morphological traits, to understand if they belong to the same taxa. In the final experimental chapter, Chapter 8, taxonomic hypothesis for Monostromataceae were tested and the systematic position of this family is explored using multi-local phylogeny. Relationships of this family with over 40 Ulvophycean genera were investigated and phylogeny reconstruction was conducted using five independent genetic markers; viz., nrDNA ITS1, nrDNA ITS2, nrDNA 5.8S, nrDNA 18S and cpDNA rbcL. Chapter 9 summarizes and discusses the results of this thesis, places them in a regional context and discusses avenues of future work.Item Monostroma: the Jeweled Seaweed for Future(2011) Bast, FelixItem Augmented anticancer efficacy of doxorubicin-loaded polymeric nanoparticles after oral administration in a breast cancer induced animal model(2011) Jain, A.K.; Swarnakar, N.K.; Das, M.; Godugu, C.; Singh, R.P.; Rao, P.R.; Jain, S.The present investigation reports an extensive evaluation of in vitro and in vivo anticancer efficacy of orally administered doxorubicin-loaded poly(lactic-co-glycolic acid) (PLGA) nanoparticles (Dox-NPs) in a breast cancer induced animal model. Spherically shaped Dox-NPs were prepared with an entrapment efficiency and particle size of 55.40 ? 2.30% and 160.20 ? 0.99 nm, respectively, and freeze-dried with 5% trehalose using stepwise freeze-drying. Cytotoxicity, as investigated on C127I cell line, revealed insignificant differences between the IC 50 of free Dox and Dox-NPs treated cells in the first 24 h, while higher cytotoxicity was demonstrated by Dox-NPs, following 72 h of incubation. Confocal laser scanning microscopy (CLSM) imaging corroborated that nanoparticles were efficiently localized into the nuclear region of C127I cells. The cellular uptake profile of Dox-NPs revealed both time and concentration dependent increases in the Caco-2 cell uptake as compared to the free Dox solution. Further, Dox-NPs significantly suppressed the growth of breast tumor in female Sprague-Dawley (SD) rats upon oral administration. Finally, orally administered Dox-NPs showed a marked reduction in cardiotoxicity when compared with intravenously injected free Dox as also evident by the increased level of malondialdehyde (MDA), lactate dehydrogenase (LDH), and creatine phosphokinase (CK-MB) and reduced levels of glutathione (GSH) and superoxide dismutase (SOD). The reduced cardiotoxicity of orally administered Dox-NPs was also confirmed by the major histopathological changes in the heart tissue after the treatments of intravenously injected free Dox and orally delivered Dox-NPs. ? 2011 American Chemical Society.Item Intracellular delivery of redox cycler-doxorubicin to the mitochondria of cancer cell by folate receptor targeted mitocancerotropic liposomes(2012) Malhi, Sarandeep Singh; Budhiraja, Abhishek; Arora, Sumit; Chaudhari, Kiran R.; Nepali, Kunal; Kumar, Raj; Sohi, Harmik; Murthy, Rayasa S.R.Cancer cells reflect higher level of ROS in comparison to the normal cell, so they become more vulnerable to further oxidative stress induced by exogenous ROS-generating agents. Through this a novel therapeutic strategy has evolved, which involves the delivery of redox cycler-doxorubicin (DOX) to the mitochondria of cancer cell where it acts as a source of exogenous ROS production. The purpose of this study is to develop a liposomal preparation which exhibits a propensity to selectively target cancer cell along with the potential of delivering drug to mitochondria of cell. We have rendered liposomes mitocancerotropic (FA-MTLs) by their surface modification with dual ligands, folic acid (FA) for cancer cell targeting and triphenylphosphonium (TPP) cations for mitochondria targeting. The cytotoxicity, ROS production and cell uptake of doxorubicin loaded liposomes were evaluated in FR (+) KB cells and found to be increased considerably with FA-MTLs in comparison to folic acid appended, mitochondria targeted and non-targeted liposomes. As confirmed by confocal microscopy, the STPP appended liposomes delivered DOX to mitochondria of cancer cell and also showed higher ROS production and cytotoxicity in comparison to folic acid appended and non-targeted liposomes. Most importantly, mitocancerotropic liposomes showed superior activity over mitochondria targeted liposomes which confirm the synergistic effect imparted by the presence of dual ligands - folic acid and TPP on the enhancement of cellular and mitochondrial delivery of doxorubicin in KB cells. ? 2012 Elsevier B.V. All rights reserved.Item Temperature-stratified screening of chickpea (Cicer arietinum L.) genetic resource collections reveals very limited reproductive chilling tolerance compared to its annual wild relatives(2012) Berger, J.D.; Kumar, Sanjeev; Nayyar, H.; Street, K. A.; Sandhu, J. S.; Henzell, J. M.; Kaur, J.; Clark, H.C.Low reproductive chilling tolerance in chickpea impairs ovule fertilization, delaying pod set, exposing the crop to terminal drought throughout much of its distribution range. Despite this realization, little progress has been made because of the limited genetic variation available to breeders. To address this issue a wide range of domesticated (n= 1762) and wild Cicer (n= 200) germplasm collected from sites stratified by flowering phase temperature was extensively field evaluated, and compared with Lupinus angustifolius, a well-adapted Mediterranean winter annual. Chilling tolerance was estimated by regressing the time interval between pod set and first flower against mean post-anthesis temperature. Field screening was augmented by smaller scale experiments evaluating the effects of contrasting post-anthesis temperature regimes on plant growth and productivity, pollen function and subsequent pod set in temperature-controlled cabinets.Chickpea was less chilling tolerant than its wild relatives, the flower-pod interval increasing curvi-linearly as sites became cooler, with a strong effects between 11 and 16. ?C, tailing off after 17.5. ?C, but remaining statistically significant. There is little useful variation for chilling tolerance within domesticated chickpea. Small, albeit statistically significant differences in pod set delay in chickpea collected from contrasting flowering phase habitats, were marginal compared to more tolerant species such as Cicer bijugum, Cicer judaicum and L. angustifolius, and to a lesser extent Cicer reticulatum, Cicer pinnatifidum, and Cicer echinospermum. No differences were observed between desi and kabuli types. Field screening identified robust chilling tolerance in a C. echinospermum accession that commenced podding earlier, at lower temperatures (10.0. ?C), and yielded 5 times more than Rupali, the most productive chickpea. Controlled temperature experiments confirmed that in contrast to chickpea, pollen germination, viability, frequency on the stigma surface and subsequent pod set were unaffected by low post-anthesis temperatures (13/7. ?C) in C. echinospermum and L. angustifolius. Our results indicate that chickpea is even more chilling sensitive than previously thought. Because C. echinospermum is inter-fertile with chickpea, it has considerable potential both as a donor of robust chilling tolerance and as an agent for investigating resistance mechanisms. ? 2011.Item Chromosomal location of non-hypersensitive leaf rust resistance genes in bread wheat cultivar PBW65 using microsatellite markers(2012) Khan, M.A.; Kamaluddin, Saini, R.G.Microsatellite or simple sequence repeat (SSRs) markers have been powerful tool for genetic mapping in wheat. Indian bread wheat (Triticum aestivum L.) cultivar PBW65 has shown significant level of resistance to most virulent race 77-5 of leaf rust (Puccinia triticina). It has been indicated that PBW65 expresses non-hypersensitive type of resistance against race 77-5. F2 and F3 crossing of PBW65 with WL711, a leaf rust susceptible wheat cultivar, and allelic tests with such already known genes (present in cultivars RL 6058 and HD 2009) revealed that cultivar PBW65 could be a potential source of novel nonhypersensitive leaf rust resistance genes. So far, only non-hypersensitive leaf rust resistance gene Lr34 was found to be effective under Indian conditions. Attempts to locate such durable leaf rust resistance genes in PBW65 through microsatellite markers showed 2B, 2D and 3D as critical chromosomes for PBW65. The primer Xgwm341 (3D) was found located 41.5 cM away from gene LrPBW1 in PBW65.Item Insulin resistance induces a segmental difference in thoracic and abdominal aorta: Differential expression of AT1 and AT2 receptors(2012) Karpe, P.A.; Gupta, J.; Marthong, R.F.; Ramarao, P.; Tikoo, K.Objectives: The study was pursued to understand and compare the vascular reactivity to angiotensin II (Ang II) and its receptor expression in thoracic and abdominal aorta under insulin resistance. Methods: Vascular reactivity to Ang II was recorded isometrically, AT1/AT2 receptor gene and protein expression was checked by RT-PCR and western blotting, respectively, and abundance of phospho (serine-10 Ph) H3 on promoter regions of Agtr1/Agtr2 genes was done by chromatin immunoprecipitation assay in aortic rings isolated from high fat diet (HFD)-fed rats. Results: Our functional studies showed an increased (Emax in mg/mm 2: Con: 319 ? 29 and HFD: 1095 ? 72, P < 0.001) and unaltered (E max in mg/mm 2: Con: 299 ? 29 and HFD: 350 ? 20, mean ? SEM, n = 6) Ang II-induced contractile responses in thoracic and abdominal aorta of HFD rats, respectively, as compared to control rats. Interestingly, AT2R-mediated relaxation was increased in abdominal aorta (% relaxation: Con: 25 ? 5.3 and HFD: 76.4 ? 8.9, P < 0.001) of HFD rats but not in thoracic aorta (% relaxation: Con: 25 ? 5.2 and HFD: 32 ? 5.2, mean ? SEM, n = 6). At the molecular level, increased mRNA (?14-folds) and protein expression (?2.5-folds) of AT2R in abdominal aorta of HFD rats was found as compared to control rats. However, AT1R mRNA and protein expression did not show any change. Chromatin immunoprecipitation with phospho H3 showed increased abundance of ser-10 phosphorylation on Agtr1 and Agtr2 gene promoter regions in thoracic and abdominal segments, respectively. But it got decreased on Agtr2 and Agtr1 genes promoter regions in thoracic and abdominal segments, respectively. Conclusion: We provide first evidence that insulin resistance induces segmental difference in thoracic and abdominal aorta and this may provide reason of heterogeneity for incidence of aneurysms. ? 2011 Wolters Kluwer Health | Lippincott Williams & Wilkins.Item Cellular uptake, intracellular trafficking and cytotoxicity of silver nanoparticles(2012) Singh, R.P.; Ramarao, P.Silver nanoparticles (Ag NPs) are used in consumer products and wound dressings due to their antimicrobial properties. However, in addition to toxic effects on microbes, Ag NPs can also induce stress responses as well as cytotoxicity in mammalian cells. We observed that Ag NPs are efficiently internalized via scavenger receptor-mediated phagocytosis in murine macrophages. Confocal and electron microscopy analysis revealed that internalized Ag NPs localize in the cytoplasm. Ag NPs cause mitochondrial damage, induce apoptosis and cell death. These effects were abrogated in presence of Ag ion-reactive, thiol-containing compounds suggesting the central of Ag ions in Ag NP toxicity. Quantitative image analysis revealed that intracellular dissolution of Ag NPs occurs about 50 times faster than in water. In conclusion, we demonstrate for the first time that Ag NPs are internalized by scavenger receptors, trafficked to cytoplasm and induce toxicity by releasing Ag ions. ? 2012 Elsevier Ireland Ltd.Item Conflict and peace in Eurasia(Taylor and Francis, 2012) Mahapatra, D.A.Focusing on a range of Eurasian conflicts, including Nagorno-Karabakh, South Ossetia and Abkhazia, this book offers contemporary perspectives on the ongoing conflicts in the Eurasia, with an emphasis on the attempts towards peace.The book brings into focus how various factors such as ethnicity, religion, border disputes, resources, and animosities inherited from the past play crucial role in these conflicts. it questions whether developments in Eurasia affect other conflicts across the globe, and if differences between parties can be resolved without pulling the relations beyond adjustable limits. The book goes on to look at how tricky the path to peace would be, and furthers the development of a framework of study of Eurasian conflicts in the post-Soviet world, while taking into account both internal and external variables in analyzing these conflicts. it is a useful contribution to Central Asian and Caucasian Politics and Security Studies. ? 2013 Selection and editorial matter, Debidatta Aurobinda Mahapatra.Item Phytogenetic analysis of bacterial diversity of heavy metal affected(Central University of Punjab, 2012) Kumar, Rajeev; Kumar, SanjeevThe 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.Item Cytotoxic activity of saragassum wightii on pc-3 cancer cell-line(Central University of Punjab, 2012) Jahid, Mohd.; Bast, FelixFinding 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.Item 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, SanjeevCancer 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.Item To study the effect of insulin on advanced androgen-indpendent prostate cancer (pc-3) cells(Central University of Punjab, 2012) Kumar, Abhimanyu; Kumar, SanjeevProstate 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.Item Histopathological staging in putative prostate cancer tissues and reviewing litearture of correlation between prostate specific antigen levels and prostate cancer inci. 2012.(Central University of Punjab, 2012) Thakur, Shweta; Thakur, SanjeevProstate cancer (PCa) remains the most significant cause of cancer specific mortality in elderly men. Asymptomatic behavior, non-modifiable risk factors and metastatic nature is the main problem of PCa. It remains clinically silent and presents itself only at advanced stage. Thus, diagnosing PCa at an early stage can result in increased chances of better treatment and hence, increased survival rate. An accurate biomarker can detect the cancer at an early stage and hence, at curable stage. Clinical parameters can only suspect prostate cancer. Whereas, histopathological examination can establish definite diagnosis of PCa. Various histological patterns are associated with cancer aggressiveness. Therefore, better understanding of clinical relevance of these histopathological findings can help to evaluate a robust biomarker for early detection of PCa. Present study was divided into two parts. First part was aimed to study the histopathology of putative prostate cancer tissue specimens. In second part, the literature of association of pre-operative serum prostate specific antigen levels with cancer detection and aggressiveness was reviewed. Protocol for histopathology of prostate tissues was established. Results of histopathological findings in putative PCa specimens were evaluated. Prevalence of histological PCa was not found in putative PCa tissues. Image library of results of the study was prepared for future analysis. Review of literature of correlation of serum PSA levels with PCa incidence suggests that PSA screening for PCa is a two-sided debate. No doubt that PSA holds the probability of detecting early PCa before development of symptoms; certain v limitations are associated with it. First, it is not reported to be 100% accurate. Second, it generates false positive and false negative results. A false positive result leads to over-treatment whereas a false negative result generates false sense of security against PCa in patient, both affects quality of life. Another main concern with PSA screening is its inability to differentiate between indolent and aggressive cancer. Therefore; accurate and economical molecular biomarkers for early detection and distinction of indolent versus aggressive cancer are urgently required. Until such biomarkers are developed and more convincing evidences regarding efficacy of PSA screening becomes available, research should focus on improving the diagnostic clinical utility of PSA by utilizing novel PSA isoforms. Identifying and validating correlation of serum PSA with tissue PSA and histological grade would be beneficial in terms of requirement of less invasive diagnostic methods to be used to measure PSA expression level as well as to confirm PCa. Future research may focus on evaluating the histological expression of other putative biomarkers and comparative serum proteomic profiling in different PCa stages.Item MicroRNA target prediction and comparative micromics; Study different cancers- special focus on breast cancer metastasis(Central University of Punjab, 2012) Sarkar, Bibekananda; Bast, FelixCancer or carcinoma is uncontrolled growth of abnormal cells. The transition of normal cell to cancerous cell is very complicated process and the cause of transition varies with situation. Metastasis is the main cause of death in cancer. There are genes which not only promote metastasis but also maintain microenvironment of tumor cells, and initiate the process of epithelium' mesenchymal transition (EMT). MicroRNA (micro RNA) is small, highly conserved noncoding RNAs that control gene expression post-transcriptionally. MicroRNA controls various cellular events like division, differentiation and apoptosis. Their deregulation may result in to cancerous growth. Most (50%) of the microRNA genes are located in the fragile chromosomal regions, which are more susceptible to amplification, deletion or translocation during tumor development. It is predicted that 30% of the all mRNA are directly or indirectly controlled by the microRNA. MetastamiR are that microRNA which have role in regulation of metastasis. MicroRNA expression profile in different cancers showed that it can act as Oncogene as well as Tumor suppressor gene. Till date there are 1921 mature human microRNA sequences registered in miRBase. MicroRNA target prediction is the first step in functional analysis of microRNA. Target prediction is complicated due to partial complementarity between microRNA and its target. There are many target prediction programs available, but the efficiency and sensitivity of these programs are not known. To enhance its efficiency, we need to know problems during prediction. Comparative analysis of different microRNA prediction tools provides an insight into the above parameter. In this study comparative analysis of seven prediction tools is carried out with help of validated microRNA targets of metastatic breast cancer.Item Effect of metform on insuln mediated proliferation of lung cancer cell lines(Central University of Punjab, 2012) Seema; Bast, FelixBackground: The incidence of cancer in diabetes mellitus (DM) patients is a matter of concern. Lung cancer is the most commonly diagnosed cancer and leading cause of death in males. Smoking is the main risk factor contributing to lung cancer progression. The occurrence of cancer is more with the type 2 DM. Besides, hyperglycemia and endogenous insulinaemia exist together for a long duration as a result of insulin resistance. As a consequence of this, the mitogenic activity of insulin is amplified. Moreover, other growth factors, and hormones are activated under the influence of insulin that further enhances this effect. It is also related to obesity, central fat accumulation, physical inactivity and smoking. The nicotine of smoke induces oxidative stress and endothelial malfunction creating metabolic abnormalities in lung cancer. In this perspective, role of insulin sensitizing drug, metformin in inhibiting the growth proliferation of lung cancer cells is hereby explored. Objective: The present study was aimed to evaluate the growth proliferation effect of insulin on non small cell lung carcinoma cell lines. It also proposed to evaluate role of metformin in preventing insulin mediated proliferation in p53 and liver kinase B1 (LKB1) mutant and wild type cell lines. Materials and methods: Two non small cell lung carcinoma cell lines, A549 and H1299 (p53 and LKB1 wild type and mutant) were used to analyze the mitogenic role of insulin by incubating for 24 hours with human recombinant insulin at a range of concentrations from 1nM to 10'M. This was followed by the metformin (concentrations from 1'M to 50mM) treatment for 24 hours along with insulin (500'M for A549 and 1mM for H1299). The proliferations were assessed by MTT dye reduction test and the percentage of the survival of the treated cells was compared with the control. One way ANOVA was used for the data analysis and the proliferation between cell lines were evaluated by student's t-test and two way analysis of variance (Two way ANOV). Results: Both the cell lines exhibited a significant proliferation (p<0.001) with the concentrations of insulin. Insulin stimulated the proliferation approximately by two fold and 1.78 times for A549 cells and H1299 cell line respectively compared to control cells. The growth of two lung carcinoma cell lines were significantly (p<0.001) inhibited by metformin treatment for 24 hours. The maximum reduction in growth was 73% and 67% for A549 and H1299 respectively for a concentration of 50mM of metformin compared to the control. The results followed a dose dependant response pattern for insulin as well as metformin treatment. Concentration at which 50% inhibition of growth observed (IC50) was comparable for both the cell lines. Conclusions: Insulin in high circulating concentrations can augment the growth proliferation of lung cancer cells. Metformin can inhibit this insulin mediated proliferation of lung cancer cells in a multifaceted way. The mechanism of action is independent of p53 and liver kinase B1.Item Effect of natrium fluoride on growth and physiology of oryza sativa L.(Central University of Punjab, 2012) Das, Anamika; Mittal, SunilThe present study was undertaken to investigate the effect of natrium fluoride on growth and physiology of Oryza sativa L. Two varieties of the O. sativa (rice) viz. Basmati 1121 and PR147 were selected for the study. PR147 is a local variety of the region. The effect was studied on growth (root length and shoot length), physiological parameters (chlorophyll content and percent respiration), macromolecules and their hydrolyzing enzymes (carbohydrate and protein content, ?-amylase and protease activity) and antioxidant enzymes (peroxidase and superoxide dismutase) under controlled conditions in the laboratory. Studies revealed that NaF (in different concentrations ranging from 16, 32 and 64 mg/l) affected the growth and physiology of both varieties as compared to control. When the seed of both varieties of rice were exposed to 64 mg/l NaF concentration, the seedling growth of test plants was highly affected as compared to control. Basmati 1121 variety was found to be more sensitive than PR147. In chlorophyll content, the percent reduction at 64 mg/l of NaF was 62% in Basmati 1121 and 30% in PR147. Similarly, reduction in percent cellular respiratory ability on exposure to 64 mg/l of NaF was 75% and 22% in Basmati 1121 and PR147 variety respectively. At 64 mg/l the carbohydrate content was increased 2.9 folds in Basmati 1121 and 1.35 folds in PR147. However the protein content was variably affected as it was increased 2 folds in PR147 and decreased to nearly 55% in Basmati 1121. The macromolecular contents were affected as the activity of their hydrolyzing enzymes was also affected. The increase in oxidative stress due to high concentrations of fluoride exposure has lead to an increase in antioxidant enzyme activity of both peroxidase and superoxide dismutase enzymes. The study concludes that high content of fluoride affects the growth and physiology of O. sativa L. by inducing oxidative stress.Item Systematics and Taxonomic Keys for the Marine Green Algal Family Monostromataceae(Nova Science Publishers, 2012) Bast, FelixMarine green algal family Monostromataceae consists of single cell-layered green seaweeds distributed throughout the world, yet identification of it being one of the most challenging. Monostromatic green seaweeds are all edible and are commercially cultivated in Japan for centuries. A thorough understanding of its systematics is indeed a necessity for any empirical investigation. Working dichotomous taxonomic key for this family is presented for the first time for the ready benefit of field phycologists. Recent advances in the systematics of this family with a special emphasize on the Japanese varieties are also presented.Item Cancer phylogenetics: Computational modeling of tumor evolution(Nova Science Publishers, Inc., 2012) Bast, FelixThe field of phylogenetics is one of the core areas of Bioinformatics which deals with computational methods to infer evolutionary heritage of organisms and genes. While phylogenetics has been extensively utilized in taxonomy and systematics of organisms, it is only very recently that the system started expanding to other fields- most importantly in cancer biology where it profoundly transformed our understanding of clonal evolution. Many of our findings in cancer phylogenetics credit to the fact that the tumor is not merely a collection of transformed cells with random mutation events; rather it is an evolving population. Many of the facets underpinning modern evolutionary synthesis can be applied to classify cancers and track its progression from initiating somatic mutation to symptomatic neoplasm. It is now widely accepted that all sub-clones within cancer are phylogenetically related and probability of a particular sub-clone progressing into neoplasm depending upon its time of initiation and evolutionary fitness. Computational models of tumor evolution have also contributed in identifying common clades- "cancer sub-types"- associated with particular cancers in different patients that in turn helped in translating our understanding of oncogeny to the development of "targeted therapeutics"- rationally designed drugs that are molecularly targeted to particular sub-types. Advent of next generation ultra-deep genome sequencing technologies has been rapidly trans-forming the very landscape of cancer phyogenetics. This chapter introduces the concept of cancer phylogenetics and reviews some of the recent advances in this field. This chapter also summarizes various phylogenetic approaches including distance matrix methods, parsimony, maximum likelihood, Bayesian methods and probabilistic inference that have potential applications in cancer research. ? 2012 Nova Science Publishers, Inc. All rights reserved.Item Sequence Similarity Search, Multiple Sequence Alignment, Model Selection, Distance Matrix and Phylogeny Reconstruction(Nature, 2013) Bast, FelixThis is a generic sequence analysis protocol suitable for plant and algal phylogeographic studies. Generated sequences from bidirectional Sanger sequencers are first assembled using Geneious. Sequence assembly is then trimmed and similarity search is conducted using BLASTn within Geneious. BLAST hits and other target taxa are selected and multiple sequence alignment is constructed. The alignment is then refined by checking using eye and exported as .fasta. Using MEGA, best-fitting nucleic acid substitution models will be calculated in MLModelTest. Model with lowest BIC score is selected and used for further phylogenetic analysis using MEGA or Geneious, which include distance matrix construction, phylogeny reconstruction using ML and BI.