VDR Potentiation and NMDA R Inhibition Facilitates Axo-Dendritic Process Formation in Melanocyte Model for Pigmented Cells in Parkinsonism

Background: A major cellular change in dopaminergic neurons leading to Parkinsonism is the alteration of microtubule proteins that causes accumulation of tau protein, α-syn and β-amyloid plaque in the cells. In this study we investigate the role of Vitamin D 3 in relieving the symptoms of Parkinsonism as it is capable of stimulating polymerization of microtubules. The Microtubules (MT) system in the fish scale melanocytes has been modeled for the dopaminergic neurons of the Substantia Nigra (SN). These cells are capable of forming cellular processes similar to what is seen in the dopaminergic neurons; in this study, we investigate the protective effect of Vitamin D 3 Receptor Agonist (VDRA) and N-Methyl-D-Aspartate Receptor (NMDA R) inhibition in process formation, synaptic denervation and melanin loss in fish scale melanocytes modeled as pigmented adrenergic cells. Method: The Tilapia scale was isolated and sub cultured in Ringer’s solution following which the cells were prepared for imaging. We incubated the cells with VDRA, Ketamine and a combination of Ketamine and VDRA in separate set ups for 60 minutes. Using brightfield imaging techniques, the cells were viewed during the incubation period and recorded using a Cameroscope connected to a computer interface. Results/Conclusion:The cells incubated with VDRA and NMDA R inhibitor, showed an increase in the number of process and extent of the process formation; the increased number of process is an indication of a rapid rate of polymerization of microtubules. Also, the processes formed are combined long processes peculiar to the NMDA R1 inhibition and short processes characteristic of VDR potentiation as seen in VDRA treatment only. Most of the effects of the VDRA were restricted to process formation around the cell body; this is similar to the microtubule cytoskeletal system found in the dendritic nucleation assembly. This finding confirms the presence of VDR and its likely restriction to d cell body plus its role in facilitating short dendrite-like process formation while NMDA R is located on the processes and facilitates long process formation

Nicotine-Cadmium Interaction Alters Exploratory Motor Function and Increased Anxiety in Adult Male Mice

In this study we evaluated the time dependence in cadmium-nicotine interaction and its effect on motor function, anxiety linked behavioural changes, serum electrolytes, and weight after acute and chronic treatment in adult male mice. Animals were separated randomly into four groups ofn= 6 animals each. Treatment was done with nicotine, cadmium, or nicotine-cadmium for 21 days. A fourth group received normal saline for the same duration (control). Average weight was determined at 7-day interval for the acute (D1-D7) and chronic (D7-D21) treatment phases. Similarly, the behavioural tests for exploratory motor function (open field test) and anxiety were evaluated. Serum electrolytes were measured after the chronic phase. Nicotine, cadmium, and nicotinecadmium treatments caused no significant change in body weight after the acute phase while cadmium-nicotine and cadmium caused a decline in weight after the chronic phase. This suggests the role of cadmium in the weight loss observed in tobacco smoke users. Both nicotine and cadmium raised serum Ca 2+ concentration and had no significant effect on K + ionwhencomparedwith the control. In addition, nicotine-cadmium treatment increased bioaccumulation of Cd 2+ in the serum which corresponded to a decrease in body weight, motor function, and an increase in anxiety

Basic Principles of Fluorescence Microscopy

Fluorescence microscopy is a basic requirement in cell biology , molecular biology and biotechnology . Advancements over the years has helped scientist to trace molecules in live cells and understand the basis of cell metabolism, exchange, mutation and to xicity. In this short communication we seek to explain in simple terms the basic principles of how a fluorescence microscope works. The principles o f excitation and emission focuses on the ability of f luorophores to absorb energy from photons and to emit such absorbed energy. The difference between t he chemical structures of these fluorephores determines how much energy that is required to exci te them and how long a fluorescence signal from a fluorophore will last. The principles of epi-illumi nation on the other hand describe the arrangement a nd function of the various components of a fluorescenc e microscope

Light microscopic detection of Plasmodium falciparum in vitro through Pf histidine rich protein 2 (HRP 2) gold conjugate labeling: Rapid diagnosis of cerebral malaria in humans

Plasmodium falciparum (Pf) has been found to be the deadliest of all the known species of the parasite capable of infecting humans; this is because it is capable of causing severe cerebral tissue damage. This study was carried out to demonstrate the parasite in the host blood in vitro through immunogold labeling using antibodies against Plasmodium falciparum histidine rich protein 2 (HRP 2); a major metabolite released during the cause of the parasite infection and feeding in the erythrocyte. 12 known Pf positive samples were obtained from across the six geopolitical zones of Nigeria and were further characterized by Geimsa thick and thin film for parasite identification parasite count expressed as parasites/l of blood. An average of 400 parasites/l of blood was obtained in each of the samples used for this study. Pf-HRP 2 antibody was conjugated to freshly prepared colloidal gold of particle size 40nm. The conjugation process was blocked with bovine serum albumin (BSA) and the conjugate itself preserved by 1% glycerol and 0.01% sodium azide. The parasite count was titrated against the Pf-HRP 2gold conjugate and was analyzed under the light microscope with a fluorescent filter. Reactivity and specificity of Pf-HRP 2 gold conjugate was found to be highly specific and gave direct identification of the erythrocytes infected with the parasite. A good contrast was also obtained between uninfected erythrocytes, parasite and the infected erythrocytes

P53, Bax and Cathepsin D Dysregulation in Neurons Subjected to Cyanide Toxicity and Oxygen Deprivation

Cyanide is a potent neurotoxin capable of potentiation NMDA R1 (N-methyl-D-aspartate receptor 1) a form of glutamate receptor that is calcium gated, thus causing excitotoxicity. It is also well established that the glutamate-glucose exchange is dependent on the activity of the Na + /K + ATPase pump, thus we examine the role of the Na + /K + pump in the metabolism of the neuron during cyanide toxicity. Six separate perfusion set up of the rat brain cortical tissues were made with ACSF (ACSF, ACSF+KCN, ACSF+KCN + pump blocker, ACSF + pump blocker). The tissues were perfused for duration of 180 minutes. The tissues were processed immunohistochemically using antibodies against p53,Bax and Cathepsin D (CD) to demonstrate disregulation of cell cycle proteins associated with the induced DNAbreakage as a result of cyanide toxicity. The pumpblockers (methyldigoxin and promethazine) induced excitotoxicity when used in culture, and amplified cyanide toxicity when combined with KCN. Cell death induced by toxicity of cyanide and the blockade of the Na/K ATPasepump has been seen to be complimentary in driving the toxicity effects that drives the cell into apoptosis.The tumor suppressor/apoptosis inducing factors p53 and Bax were over expressed while cathepsin was suppressed to show that the cells are apoptotic as against an increased cathepsin D level that would have implied senescence