Dopamine Receptor Subtypes,IP3, cAMP and cGMP Functional Regulation in Parkinsonism Induced by Unilateral Infusion of Rotenone in Rats: Recovery with Bone Marrow Cells,Serotonin and GABA Supplementation

In the present study, the effects of 5-HT, GABA and Bone Marrow Cells infused intranigrally to substantia nigra individually and in combinations on unilateral rotenone infused Parkinsonism induced rats. Scatchard analysis of DA, DA D1 and D2 receptors in the corpus striatum, cerebral cortex, cerebellum, brain stem and hippocampus showed a significant increase in the Brain regions of rotenone infused rat compared to control. Real Time PCR amplification of DA D1, D2, Bax and ubiquitin carboxy-terminal hydrolase were up regulated in the brain regions of rotenone infused rats compared to control. Gene expression studies of -Synuclien, cGMP and Cyclic AMP response element-binding protein showed a significant down regulation in Rotenone infused rats compared to control. Behavioural studies were carried out to confirm the biochemical and molecular studies.Our study demonstrated that BMC administration alone cannot reverse the above said molecular changes occurring in PD rat. 5-HT and GABA acting through their specific receptors in combination with bone marrow cells play a crucial role in the functional recovery of PD rats. 5-HT, GABA and Bone marrow cells treated PD rats showed significant reversal to control in DA receptor binding and gene expression. 5-HT and GABA have co-mitogenic property. Proliferation and differentiation of cells re-establishing the connections in Parkinson's disease facilitates the functional recovery. Thus, it is evident that 5-HT and GABA along with BMC to rotenone infused rats renders protection against oxidative, related motor and cognitive deficits which makes them clinically significant for cellbased therapy. The BMC transformed to neurons when co-transplanted with 5-HT and GABA which was confirmed with PKH2GL and nestin. These newly formed neurons have functional significance in the therapeutic recovery of Parkinson’s disease.

Bone marrow cells differentiation to neurons in the rat brain using Serotonin and GABA:Their role on glutamate receptors, IP3, cAMP and cGMP functional regulation in unilateral Parkinsonism induced by 6-hydroxydopamine

Parkinson's disease is a chronic progressive neurodegenerative movement disorder characterized by a profound and selective loss of nigrostriatal dopaminergic neurons. Our findings demonstrated that glutamatergic system is impaired during PD. The evaluations of these damages have important implications in understanding the molecular mechanism underlying motor, cognitive and memory deficits in PD. Our results showed a significant increase of glutamate content in the brain regions of 6- OHDA infused rat compared to control. This increased glutamate content caused an increase in glutamatergic and NMDA receptors function. Glutamate receptor subtypes- NMDAR1, NMDA2B and mGluR5 have differential regulatory role in different brain regions during PD. The second messenger studies confirmed that the changes in the receptor levels alter the IP3, cAMP and cGMP content. The alteration in the second messengers level increased the expression of pro-apoptotic factors - Bax and TNF-α, intercellular protein - α-synuclein and reduced the expression of transcription factor - CREB. These neurofunctional variations are the key contributors to motor and cognitive abnormalities associated with PD. Nestin and GFAP expression study confirmed that 5-HT and GABA induced the differentiation and proliferation of the BMC to neurons and glial cells in the SNpc of rats. We also observed that activated astrocytes are playing a crucial role in the proliferation of transplanted BMC which makes them significant for stem cell-based therapy. Our molecular and behavioural results showed that 5-HT and GABA along with BMC potentiates a restorative effect by reversing the alterations in glutamate receptor binding, gene expression and behaviour abnormality that occur during PD. The therapeutic significance in Parkinson’s disease is of prominence.

Strength Predictions of Hat Stiffened Plate

Hat Stiffened Plates are used in composite ships and are gaining popularity in metallic ship construction due to its high strength-to-weight ratio. Light weight structures will result in greater payload, higher speeds, reduced fuel consumption and environmental emissions. Numerical Investigations have been carried out using the commercial Finite Element software ANSYS 12 to substantiate the high strength-to-weight ratio of Hat Stiffened Plates over other open section stiffeners which are commonly used in ship building. Analysis of stiffened plate has always been a matter of concern for the structural engineers since it has been rather difficult to quantify the actual load sharing between stiffeners and plating. Finite Element Method has been accepted as an efficient tool for the analysis of stiffened plated structure. Best results using the Finite Element Method for the analysis of thin plated structures are obtained when both the stiffeners and the plate are modeled using thin plate elements having six degrees of freedom per node. However, one serious problem encountered with this design and analysis process is that the generation of the finite element models for a complex configuration is time consuming and laborious. In order to overcome these difficulties two different methods viz., Orthotropic Plate Model and Superelement for Hat Stiffened Plate have been suggested in the present work. In the Orthotropic Plate Model geometric orthotropy is converted to material orthotropy i.e., the stiffeners are smeared and they vanish from the field of analysis and the structure can be analysed using any commercial Finite Element software which has orthotropic elements in its element library. The Orthotropic Plate Model developed has predicted deflection, stress and linear buckling load with sufficiently good accuracy in the case of all four edges simply supported boundary condition. Whereas, in the case of two edges fixed and other two edges simply supported boundary condition even though the stress has been predicted with good accuracy there has been large variation in the deflection predicted. This variation in the deflection predicted is because, for the Orthotropic Plate Model the rigidity is uniform throughout the plate whereas in the actual Hat Stiffened Plate the rigidity along the line of attachment of the stiffeners to the plate is large as compared to the unsupported portion of the plate. The Superelement technique is a method of treating a portion of the structure as if it were a single element even though it is made up of many individual elements. The Superelement has predicted the deflection and in-plane stress of Hat Stiffened Plate with sufficiently good accuracy for different boundary conditions. Formulation of Superelement for composite Hat Stiffened Plate has also been presented in the thesis. The capability of Orthotropic Plate Model and Superelement to handle typical boundary conditions and characteristic loads in a ship structure has been demonstrated through numerical investigations.