Botany - Mphil Thesis

Permanent URI for this collectionhttps://kr.cup.edu.in/handle/32116/30

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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.
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.
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.
Effect of metform on insuln mediated proliferation of lung cancer cell lines
(Central University of Punjab, 2012) Seema; Bast, Felix
Background: 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.
MicroRNA target prediction and comparative micromics; Study different cancers- special focus on breast cancer metastasis
(Central University of Punjab, 2012) Sarkar, Bibekananda; Bast, Felix
Cancer 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.
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.
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.