Zoology - Master Dissertation
Permanent URI for this collectionhttps://kr.cup.edu.in/handle/32116/63
Browse
12 results
Search Results
Item Oxidative stress responses to sub-lethal dose of Cry toxin in the larvae of castor semilooper, Achaea janata(Central University of Punjab, 2018) Singh, Kanika; Chaitanya,R.K.Development of synthetic insecticides to reduce the level of infestation led to deleterious effects on environment and human health. This lead to the development of ecofriendly pest management alternatives including Bacillus thuringensis (Bt). Bt produce Crystal (Cry), Cytotoxic (Cyt) and Vegetative (Vip) proteins with insecticidal activity against different orders of lepidoptera. Of late, pest resistance against Bt is reported in countries.The reduced toxicity of Bt formulation from degradation by UV light, wash-off by rain, drying, temperature, and soil acidity as well as its chemistry. Further, insects sense pesticides through odorant receptors and move away quickly, there is always a possibility of a population of larvae to get exposed to sub-lethal doses of toxin which might exhibit variable effects and escape mortality and eventually generate resistance. Sub-lethal dose lead to the generation of oxidative stress in the insect and eventually scavenged by anti-oxidant enzymes. These stress responses would enhance our understanding of adaptations for survival and resistance development. The current study is an attempt to monitor the antioxidative responses at the transcriptional level upon sub-lethal exposure of Cry toxin in the larvae of an polyphagous pest castor semilooper, Achaea janata. prevalent in the Indian subcontinent.Item Isolation and characterization of platelet microvesicles(Central University of Punjab, 2018) Kumar, Kapil; Singh,Sunil KumarIn the recent years, Platelet microvesicles (PMVs) are projected as potential diagnostic and prognostic biomarkers of many diseases like cardiovascular diseases, diabetes and infection. Due to the small size (100-1000 nm), diverse structure and functions, PMVs is associated with different challenges during isolation, quantification and characterization. Hence, it is pertinent to optimize methods to isolate, characterize and quantify MVs in blood plasma. Therefore, in this study it is aimed to isolate and characterize the PMVs release upon platelet activation. The released PMVs were isolated by differential centrifugation and characterized through scanning electron microscopy (SEM) and flow cytometer (FACS). SEM images showed resting platelets as smooth spherical cells of 2- 3 ?m in diameter separated from each other having presence of small hyaloplasmic extensions. In contrast, platelet activation elicits significant morphological change exhibited well-developed pseudopods (hyaloplasmic processes), which connected the cells forming large aggregates (clumps). Electron microscopy and flow cytometry data showed platelet microvesicles release of size in the range of 200-500 nm from the activated platelets. Thus, in this study an isolation and characterization technique for platelet microvesicles was optimized that can be further validated with clinical samples and exploit to study microvesicles structure, function and clinical importance.Item Neuro-Protective Role Of Ginkgolide B In A?induced Neurodegeneration And Ache Enzyme Activity In Human Neuroblastoma Sh-Sy5Y Cells(Central University of Punjab, 2018) Mukherjee, Ankita; Mantha,Anil K.Ginkgolide B (GB) is being used as medicine in China for treating neurodegenerative diseases for a long time. Its neuroprotective role is getting well established. Alzheimer's disease (AD) is a neurodegenerative disease that has multiple factors associated with its onsetand is one of the most common causes of dementia in the world. GB is known to reduce the oxidative stress caused due to accumulation of amyloid beta (A?), a major hallmark of AD associated strongly with the production of oxidative stress via production of ROS. The increase in the expression of AChE has been reported and it has been associated with increased toxicity of A?. This study tried to decipher the relationship between A?, GB and AChE activity. In this study, it was found that A?(25-35)-induced oxidative stress leads to increased production of ROS and decreased AChE activity. On the other hand, GB decreased ROS production and expression of AChE, thus pointing toward its protective effect. GB increased the activity of AChE, suggesting that due to its antioxidant potentialit possibly caused a decrease in protein oxidation, and thus increased the activity of the AChE enzyme. Therefore, the results of the present study show the modulatory role of GB an AChE enzyme activity under oxidative stress conditions as seen in AD, suggesting the potential of GB in AD therapeuticsItem To Test The Role Of Withaferin A In Neuroprotection On Sh-Sy5Y Cell Line Challenged By Kainic Acid(Central University of Punjab, 2018) Pruseth, Aishwarya; Parkash, JyotiWithania somnifera (WS) has been used in traditional medicine for a variety of neural disorders. Recently, chronic neurodegenerative conditions have been shown to benefit from treatment with this crude extract. Neurotoxicity induced by Kainic acid causes stroke, head trauma and leads to various neurodegenerative diseases. It is a common pathway for neuronal injury and death. Withaferin A (WA) is the steroidal lactone and phytochemical of medicinal plant WS. These days research on the medicinal plant is the major theme so as to identify the novel, natural and safe phytotherapies has been flourished. WS is a popular ayurvedic plant with an array of medicinal properties including neuroprotection. It also appears to exert a great influence on the cardiopulmonary, endocrine, and central nervous systems. Recently several in-vitro and in-vivo preclinical studies validating the therapeutic value of newly identified phytochemicals have been launched. The use of medicinal plants and its chemical component can be exploited in the treatment of neurodegenerative diseases like Parkinson, Alzheimer's, Epilepsy and so on. Ayurvedic products are the only promising way to treat these diseases with negligible side effects. In the current study the neuroprotective role WA has been studied on SH-SY5Y neuroblastoma cell line challenged by kainic acid. Cell viability and expression of neuronal cell differentiation marker NCAM was investigated on v Kainic acid challenged SH-SY5Y cells with and without the presence of WA. When the cells are challenged via Kainic acid there is a decrease in cell viability due to the phenomenon of excitotoxicity and this was reversed by WA treatment. NCAM is an important cell surface plasticity marker that have role in repair and regeneration. Potential mechanisms underlying the observed neuroprotection were examined. Additionally, morphological changes were monitored following KA treatment. Upon excitotoxicity with Kainic acid NCAM expression increased but the effect is reverse upon pre-treatment with WA. Altogether, the present study suggests that WA is neuroprotective and provides neuroprotection against KA induced excitotoxicity could have therapeutic potential to target factors involved in neurodegenerative diseases.Item Expression analysis of long non-coding RNA GAS5 and BANCR in lung cancer cell line A549 compared to IMR-90.(Central University of Punjab, 2018) Sharma, Uttam; Jain, AklankLung cancer is the major cause of death worldwide. Several chemotherapeutic drugs and therapies have been established, but the early diagnosis and prognosis of lung cancer is still a question. Long non-coding RNAs are important regulator molecules in the human genome, which serves as transcriptional modulator, post transcriptional processor, chromatin remodeletor and splicing regulator during the gene modification process. Emerging studies have suggested the role of long noncoding RNA as potential biomarker for cancer diagnosis and prognosis by functioning as tumor suppressors and oncogenes. Several studies have been reported on cell lines, tissues as well as tumor and the molecular mechanism is still not clearly understood. GAS5 and BANCR are two long non-coding RNAs, which are found to be down-regulated in multiple cancers such as lung carcinoma, breast cancer, etc. In the current study, we focus on the expression analysis of GAS5 and BANCR in A549 cell line compared to IMR-90 cell line to study the role of long non-coding RNA in the pathogenesis of lung cancer. Furthermore, we investigated the expression of GAS5 and BANCR using quantitative Real-Time PCR. The result showed that GAS5 and BANCR expression was significantly down-regulated in cancerous cell line compared to non-cancerous cell line. The fold change of lncRNAs GAS5 and BANCR was 14 times (P=0.0088) and 7 times (P=0.0088) down-regulated in A549 cell line respectively. The melt curve analysis showed that there was only one sharp peak obtained for both GAS5 and BANCR i iv which suggests that primers bind to their specific targets and no primer dimer was observed.Item To check the glioprotective effect of Withaferin A on C6 Glioma cell Culture challenged with Kainic Acid(Central University of Punjab, 2018) Dash, Bindu Balaya; Parkash, JyotiLast many years perception of glial cell function in CNS has been changed. In fact abnormalities in glial cells also contribute to various disorders. Glutamate induced excitotoxicity attributed to various CNS related diseases. Kainic acid is a potent agonist of kainate receptors, a subclass of glutamate receptors, is 30 fold more neurotoxic than glutamate. Natural herbal extracts are attracting researchers for their pharmacologic properties against diseases associated with CNS. Root and leaf extracts of Withania somnifera, used since many years to treat several diseases in traditional medicine system. Present study was designed to see the glioprotective potential of Withaferin A, a natural extract from Withania somnifera or Ashwagandha, against Kainic acid induced excitotoxicity in C6 glioma cell line. Pretreatment of 0.5 µM Withaferin A showed defensive potential against 100 µM and 200 µM concentration of Kainic acid. To check expression of GFAP (well known marker of astrocytes) and NCAM, Immunocytochemistry was performed. Withaferin A treatment helps in normalizing of GFAP (Glial Fibrillary Acidic Protein) and NCAM (Neural Cell Adhesion Molecule) expression in kainic acid exposed cells. Our result suggested that Withaferin A have defensive potential against kainic acid induced excitotoxicity. As a potent glioprotective agent, Withaferin A could be used as therapeutic drug to treat glioblastomas and other neurological disorders.Item To Study the Anti-Proliferative Effect of Pirfenidone via MAP Kinase Signaling Pathway(Central University of Punjab, 2018) Bharti, Pooja; Parkash, JyotiPirfenidone is a pyridine chemical compound used as a therapeutic drug used for the treatment of Idiopathic Pulmonary Fibrosis (IPF). The characteristic feature of the fibrosis is the deposition of extracellular matrix protein in the lungs and ultimate lead to the failure of the organ. Pirfenidone is an orally active small molecular comprising a modified phenyl pyridine that have been shown to inhibit the progression of fibrosis in animal model and in patient with IPF. The compound exhibits well?documented anti-fibrotic and anti? inflammatory activities but its molecular target has not been elucidated. Pirfenidone is a broad-spectrum anti-fibrotic drug with the ability to decreases the PDGF, TGF-?, TNF-? and COL (collagen). After treating A549 cell line with Pirfenidone with different concentrations and different time interval, cell viability was measured by the MTT assay. Pirfenidone cause the cytotoxicity in model A549 cell line and anti-proliferation through different Smad dependent and independent pathway. Pirfenidone treatment was given to check downstream signaling pathway i.e. MAP kinase activity dependent. Although previous studies have shown that Pirfenidone is associated with renal fibrosis in TGF??1 expression the mechanism remains poorly understood. Taken into account these, we hypothesized that pirfenidone may have an anti?fibrotic effect through antagonizing the MAPK signaling pathway.Item Role of midgut digestive proteases upon sub-lethal exposure of Cry toxin in the larvae of castor semilooper, Achaea janata(Central University of Punjab, 2018) Sureshgopi, D; Chaitanya, R.K.Development of synthetic insecticides to reduce the level of infestation led to deleterious effects on the environment and human health. This lead to the development of eco-friendly pest management alternatives including Bacillus thuringensis (Bt). Bt produces Crystal (Cry), Cytotoxic (Cyt) and Vegetative (Vip) proteins with insecticidal activity against different orders of Lepidoptera. Of late, pest resistance against Bt is being reported in many parts of the country. The foliar spray may lead to the reduced toxicity of Bt formulation from degradation by UV light, wash-off by rain, drying, temperature, and soil acidity as well as its chemistry. Further, insects sense pesticides through odorant receptors and move away quickly. Under these conditions, there is always a possibility of a population of larvae to get exposed to sub-lethal doses of toxin which might exhibit variable effects and escape mortality and eventually generate resistance. The role played by digestive proteases in the activation of the Cry toxin and digestion when administered in sub-lethal concentration could help in the elucidation of key phenomena like adaptations for survival and resistance development. Elucidation of these digestive proteases would enhance our understanding of resistance development. The current study is an attempt to monitor the transcript levels of digestive proteases upon sub-lethal exposure of Cry toxin in the larvae of an economically important polyphagous pest castor semilooper, Achaea janata prevalent in the Indian subcontinent.Item Traumatic Brain Injury (TBI) causes hypersomnia in Drosophila melanogaster(Central University of Punjab, 2018) Birua, Sunil Kumar; Lone,Shahnaz RahmanTraumatic brain injury (TBI) is one of the deadly event which is a leading cause of neurological impairments and loss of life. The TBI outcomes arise from primary and secondary injuries. It damages neurons as well as causes neuronal death in the brain. Primary injuries appear when the first jolt occurs to the brain and are prompted by external mechanical forces, which causes distortion of the brain, whereas secondary injuries are elicited by cellular and molecular responses in response to the primary injuries. The underlying mechanisms to prevent various consequences associated with TBI are yet unknown. We have designed a model of TBI in Drosophila melanogaster. The model facilitates us to trace out valuable experimental tools available in flies. The spring operated High impact trauma (HIT) device allows us to expose the flies to a rapid back and forth injuries. Drosophila activity monitor systems (DAMS) are used for the recording of activity. The TBI exposed flies shows sleep alteration compared to control groups. Flies die faster when subjected to the TBI as compared to controls. We confirmed by the two separate methods inactivity observed is sleep and is not due injury in other body parts. We further showed circadian clock may have important role to play in protecting organism from injuries as clock null flies are more susceptible to the injuries Thus our study demonstrates the potential importance of D. melanogaster as a model organism for addressing the questions related to TBI and hypersomnia.Item Development of lactadherin based electrochemical biosensor for the detection of platelet microvesicles(Central University of Punjab, 2018) Santra, Sneha; Singh, SunilPlatelet derived microvesicles (PMVs) are the most abundant membrane vesicles in the blood having a potent pro-inflammatory effect, promote coagulation and affect vascular function which are involved in the pathogenesis of cardiovascular disease including diabetes, thrombosis, and coronary artery diseases. Therefore, it is pertinent to detect PMVs level in blood of an individual which have prognostic potential for cardiovascular diseases. As per literature available, lactadherin is a small (53-66 kDa) multifunctional glycoprotein which plays an important role in the clearance of microvesicles. In the present study, lactadherin based electrochemical biosensor for the detection of PMVs was explored. Polythionine film (as a good electron mediator) was electrochemically deposited on ITO-coated glass through electrochemical process involving cyclic voltammetry (CV) and chronoamperometry. Electrochemically deposited electrode provides ideal adsorbing platform for immobilization of RGDS peptide sequences (Lactadherine binding motif) having binding affinity against active conformation of integrin on PMVs surface. CV and Diffusion Pulse Voltammetry (DPV) measurements showed gradual decrease of v current with the subsequent adsorption of microvesicles poor plasma (MPP), microvesicles rich plasma (MRP) and activated platelets. Decrease of current clearly depicts the presence of microvesicles in blood plasma. Our developed fabricated electrode can have a promising potential for its efficient application in clinical testing of various pathological conditions.