A special type of metal side disk rotating electrode has been demonstrated for the direct estimation of total arsenic [As(III) and As(V)] at low ppb (mg/L) level using anodic stripping differential pulse voltammetry (ASDPV). A conventional three electrodes cell is used equipped with side disk rotating gold electrode as working, graphite/ platinum electrode as auxiliary and Ag/AgCl/3M KCl as reference electrodes. Arsenic is estimated in various acidified samples without any digestion, containing a trace amount of copper at low ppb level. The major problems associated with ASDPV of high acidic condition (acid hazards), irreproducible results due to the interference of hydrogen bubbles, evolved at the cathode during the deposition of arsenic in acidified samples and poor detection level are overcome with the help of the specially designed gold side disk rotating electrode and modified electrolyte. The presence of a trace amount of copper(II) salt in the electrolyte is found to enhance the sensitivity of the technique. The shape and position of the metal disk at the electrode, rotation speed of the electrode and electrolyte are optimized to have less hydrogen gas bubbles interference and high reproducibility in the detection of arsenic up to 2 ppbAE 15% level in various samples. The electrode has been found very stable and reproducible even for more than 200 estimations.
Among trophic factors already known, glial cell line-derived neurotrophic factor (GDNF) and other members of its family have potent and specific action on dopaminergic neurons. In the present investigation an attempt has been made to validate the role of GDNF co-transplantation with fetal ventral mesencephalic cells (VMC) on functional viability and restoration using neurobehavioral, neurochemical and immunohistochemical parameters at 6 weeks post-transplantation in 6-hydroxydopamine (6-OHDA) lesioned rat model of Parkinson's disease (PD). A significant restoration (P<0.01) in D-amphetamine induced rotations, spontaneous and apomorphine induced locomotor activity in rats co-transplanted with VMC and GDNF was observed as compared to VMC alone transplanted rats. Level of dopamine (DA), 3,4-dihydroxy-phenyl acetic acid (DOPAC) and dopamine D2 (DA-D2) receptors in the caudate putamen (CPu) were significantly (P<0.001) restored in co-transplanted group as compared to VMC transplanted or GDNF administered animals. The functional viability of transplanted VMC was confirmed by tyrosine hydroxylase (TH) expression and quantification of TH-positive cells by image analysis revealed a significant restoration in TH-IR fibers density as well as TH-IR neurons counts in co-transplanted animals over VMC transplanted animals. Results suggest that co-transplantation of VMC and GDNF may be a better approach towards functional restoration in 6-OHDA lesioned rat model of Parkinson's disease.
The present study was undertaken to elucidate the role of circulating neutrophils if any in oxidative stress in migraine by evaluating free radical generation and activities of enzymatic antioxidants in the blood in 55 patients with migraine and 60 healthy controls. Free radical generation was assessed by flow cytometry, while activity of catalase, superoxide dismutase (SOD) and glutathione peroxidase (GPx) was estimated in blood polymorphonuclear neutrophils (PMNs) by standard procedures. Platelet SOD was also measured. No significant change was found in free radical generation and in the activity of catalase, SOD and GPx in migraine patients. Univariate analysis of PMN catalase level revealed that migraineurs with a positive family history had significantly lower catalase activity compared with those with a negative family history. No correlation was found in the activity of antioxidant enzymes with age, duration of disease, time since last attack and headache index. The platelet SOD also did not show any significant change in patients of migraine without aura. Platelet aggregation in the presence or absence of PMNs was also not altered significantly. Thus the findings of the present study suggest that neutrophils are not the cause of oxidative stress observed in migraine patients.
Exogenous administration of various neurotrophic factors has been shown to protect neurons in animal model of Parkinson's disease (PD). Several attempts are being made to search a tissue source simultaneously expressing many of these neurotrophic factors. Carotid body (CB) contains oxygen-sensitive glomus cells rich in dopamine (DA) and expresses glial cell line-derived neurotrophic factor, brain-derived neurotrophic factor and neurotrophin-3. We have attempted to study the functional restoration following co-transplantation of CB cells and ventral mesencephalic cells (VMC) in a 6-hydroxydopamine-lesioned rat model of PD. A significant recovery (p < 0.001) in D-amphetamine-induced circling behavior (80%) and spontaneous locomotor activity (85%) was evident in co-transplanted animals at 12 weeks post-transplantation as compared to lesioned animals. Similarly, a significant (p < 0.001) restoration was observed in DA-D 2 receptor binding (77%), striatal DA (87%) and 3,4-dihydroxyphenylacetic acid (DOPAC) (85%) levels and nigral DA (75%) and DOPAC (74%) levels. Functional recovery was accompanied by tyrosine hydroxylase (TH) expression and quantification of TH-positive cells by image analysis revealed a significant restoration in TH-immunoreactive (IR) fiber density in striatum, as well as TH-IR neurons in substantia nigra pars compacta in co-transplanted animals over VMC-transplanted animals. The result suggests that co-transplantation of CB cells along with VMC provides better and long-term functional restoration in the rat model of PD, possibly by supporting the survival of newly grafted cells as well as remaining host DA neurons.
Reactive dopamine (DA) metabolites have been implicated in both Parkinson's disease and manganese (Mn) neurotoxicity. Rat PC12 and genetically modified PC12 (PC12M) cells capable of producing higher DA content, on exposure to MnCl2 (10¡ 6 M) for 72 hours, exhibited a significant decrease in glutathione content. Activity of antioxidant enzyme catalase was also inhibited following 24 and 72-hour MnCl2 exposure. MnCl2 caused a concentration dependent (10¡ 7 to 10¡3 M) loss in mitochondrial activity after 24 and 72 hours and an impaired DNA synthesis after 72 hours with changes being more marked in PC12M cells. The results indicate that the free-radical-mediated toxicity of Mn at cellular level involves down-regulation of anti oxidants in normal and DA-rich PC12 cells. PC12M cells appeared to be more sensitive than PC12 cells.
A new solid-state pH sensor is developed using neutral poly(3-cyclohexyl thiophene) assembled over a Pt disk electrode. The new sensor is developed following two different approaches; 1) the neutral poly(3-cyclohexyl thiophene) dissolved in chloroform and subsequent coating on to a Pt disk electrode; 2) the neutral polymer is incorporated into plasticized poly(vinyl chloride) matrix membrane. In both cases the polymer modi®ed electrode is sensitive to pH and a reversible super Nernstian behavior is observed. The typical response of the pH sensor and its reversibility are reported. The polymer coated electrode is subsequently used to construct an all solid-state urea sensor. The construction of this new urea sensor involves the following two major steps; a) 20 mL of urease solution (40 mg /mL) is allowed to assemble overnight at 4 C over neutral poly (3-cyclohexyl thiophene) modi®ed electrode; b) an organically modi®ed sol-gel layer is allowed to form over the urease adsorbed polymer modi®ed electrode. The new solid-state urea sensor provides excellent reproducibility of the measurements and is stable for 3 months when stored at 4 C under dry condition. The typical response of the solid-state urea sensor and the calibration plot of urea analysis are reported.
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