Locomotor behavior of bonnet monkeys after spinal contusion injury: footprint study.

Analysis of gait functions following spinal cord injury has been widely studied in rats, mice but limited in primates. This investigation was performed to quantitatively analyze the degree of functional recovery in bipedal locomotion in bonnet monkeys after induced spinal cord contusion. The degree of locomotor recovery was examined by measuring four gait variables, viz., tip of opposite foot (TOF), print-length (PL), toe-spread (TS), and intermediary toe-spread (IT) from the recorded hindlimb prints of monkeys using ink and paper technique. Contusion was induced in spinal cord at T12-L1 level in anaesthetized monkeys by using the Allen's weight drop technique. Postoperatively, all spinal contused animals initially showed a significant decrease in TOF, which then gradually increased for longer duration and attained the near normal values by the sixth month. On the other hand, PL, TS, and IT variables in hindlimb prints of contused animals were found to dramatically increase initially and then slowly decrease subsequently. Later there was a recovery to insignificant levels which differed from the corresponding preoperative values by the fifth month. The observations of this study suggest that the functional contributions of the spared fibers, especially in ventral and ventrolateral funiculi, through collateral sprouts or synaptic plasticity that were formed in the contused spinal cord may be responsible for substantial recovery of hindlimb movements. Moreover, based on analysis of footprint variables observed in locomotion in these subjected monkeys, we understand that spinal automatism and development of responses by afferent stimuli from outside the cord could possibly contribute to recovery of the paralyzed hindlimbs.

Regulation of protein kinases and coregulatory interplay of S-100beta and serotonin transporter on serotonin levels in diabetic rat brain.

Protein kinases are critical component in the regulation of signal transduction pathways, including neurotransmitters. Our previous studies have shown that serotonin (5-HT) altered under diabetic condition was accompanied by alterations of protein kinase C-alpha (PKC-alpha) and CaMKII, and those alterations were reversed after insulin administration. The current study showed that alloxan-induced diabetic animals revealed hyperglycemia and was associated with an increase in the content of 5-HT, PKC-alpha expression and PKC activity (P < 0.05) simultaneously in striatum (ST), midbrain (MB), pons medulla (PM), cerebellum (CB), and cerebral cortex (CCX) from 7 days to 60 days. Although the 5-HT levels in hippocampus (HC) and hypothalamus (HT) were not altered, the PKC-alpha expression and PKC activity showed increases (P < 0.05) in level in HC. Insulin administration reversed all these changes to a normal level. In contrast, the in vitro study has shown that the 5-HT levels correlated with PKC-alpha expressions as well as PKC activity (P < 0.05) only in ST, MB, and CB either after induction with phorbol 12-myristate 13-acetate (PMA) or blocking with chelerythrine, whereas PM and CCX remained elevated (P < 0.05), implying a regulatory role for PKC-alpha only in ST, MB, and CB. However, our consecutive studies have shown that the 5-HT level in PM was regulated by p38-mitogen-activated protein kinase (p38-MAPK) both in vivo and in vitro, whereas the 5-HT level in CCX was coregulated by S-100beta by protein-protein interaction with serotonin transporter (SERT) via 8-bromoadenosine 3',5'-cyclic monophosphate sodium salt (8-Br-cAMP)-induced cAMP/PKAII pathway(s).

An acute hyperglycemia or acidosis-induced changes of indolamines level correlates with PKC-alpha expression in rat brain.

Hyperglycemia and ketoacidosis are the two most serious factors in acute metabolic complications of both type 1 and type 2 diabetes. Dysfunction of the central nervous system is a well-documented complication of diabetes. We and others have previously reported that acute or chronic diabetes in animal's results in altered brain neurotransmitter levels. In this study, we investigated the effects of acute (7 days) glucose-induced hyperglycemia and sodium acetoacetate (NaAcAc) or ammonium chloride (NH4Cl) induced acidosis on the level of indolamines (5-hydroxytryptamine (5-HT) and 5-hydroxyindoleacetic acid (5-HIAA)) as well as PKC-alpha expression/activity in discrete areas of rat brain. Glucose-induced (500 mg/kg, bw) hyperglycemic ( approximately 249 mg%) rats showed significant (p<0.05) increase in 5-HT levels in mid brain (MB), pons medulla (PM) and cerebellum (CB), respectively. 5-HIAA level increased in hippocampus (HC) (p<0.05) as compared to control. The rats treated with sodium acetoacetate (NaAcAc) for 7 days (60 mg/kg, bw) showed significant decrease (p<0.05) of 5-HT level in hypothalamus (HT). Whereas, the 5-HIAA level increased in MB (p<0.05). Similarly, the PKC-alpha expression as well as the enzyme activity showed significant increase in HC, MB, PM and CB under glucose-induced hyperglycemia and that changes correlated the changes of indolamines, suggesting that the hyperglycemia may be the major metabolic disorder in diabetic complications.

A short-term diabetes induced changes of catecholamines and p38-MAPK in discrete areas of rat brain.

Chronic diabetes is associated with the alteration of second messengers and CNS disorders. We have recently identified that protein kinases (CaMKII and PKC-alpha) and brain neurotransmitters are altered during diabetes as well as in hyperglycemic and acidotic conditions. In this study, we investigated the effects of acute diabetes on the levels of dopamine (DA), norepinephrine (NE), epinephrine (E) and p38-Mitogen-Activated Protein Kinase (p38-MAPK) in striatum (ST), hippocampus (HC), hypothalamus (HT), midbrain (MB), pons medulla (PM), cerebellum (CB) and cerebral cortex (CCX). Alloxan (45 mg/kg) diabetic untreated rats that showed hyperglycemia (>260 mg%), revealed significant increases of DA level in ST (1.5 fold), HC (2.2 fold) and PM (2.0 fold) and the E level also found to be increased significantly in HT (2.4 fold), whereas the NE level was decreased in CB (0.5 fold), after 7 days of diabetes. Immunoblotting study of p38-MAPK expression under identical conditions showed significant alterations in ST, HC, HT and PM (p<0.05) correlated with the changes of catecholamines (DA and E). On the other hand, the above changes were reversed in insulin-treated diabetic rats maintained under normal glucose level (80 -110 mg %). These results suggest that p38-MAPK may regulate the rate of either the synthesis or release of DA and E in corresponding brain areas, but not NE, under these conditions.

Effect of Ocimum sanctum Linn on the changes in central cholinergic system induced by acute noise stress.

The ethanolic extract from the leaves of Ocimum sanctum Linn was screened for its effects on the noise induced changes in the central cholinergic system of albino rats by investigating the acetylcholine content and acetylcholinesterase activity in discrete areas of brain. Exposure to noise (10 kHz:100 dB) stress for 30 min caused a significant reduction in total acetylcholine content and increase in the activity of acetylcholinesterase in cerebral cortex, corpus striatum, hypothalamus and hippocampus of brain. Pretreatment of the animals with ethanol extract of Ocimum sanctum leaves for 7 days prevented the noise induced changes in these two cholinergic parameters in all the four areas of brain. The results of the study indicate the protective nature of the plant material on the brain tissues against the detrimental effect of noise stress.

Modulation of specific immunity by ventral hippocampal formation in albino rats.

Rats with bilateral electrolytic ventral hippocampal formation (VHF) lesion show a significantly (p less than 0.001) enhanced migration of leucocytes in the presence of antigen. Immunization in these animals enhances (p less than 0.001) the leucocyte migration and presents a significant decrease in footpad thickness. Antibody titre (p less than 0.001) and plaque-forming cells (p less than 0.02) also show a significant decrease when compared with sham-lesion-immunized animals and control-immunized animals. These data indicate the influence of VHF in modulating the specific immunity.

Modulation of non-specific immunity by hippocampal stimulation.

Rats subjected to electrical stimulation of hippocampus (30 min/day for 4 days) showed an increase in the neutrophils in the peripheral blood when compared to sham (P < 0.01) and controls (P < 0.001). They also showed a significant decrease in lymphocytes, when compared to control rats (P < 0.001). Both sham and stimulated animals showed a significant decrease in total white blood cell count when compared to controls (P < 0.001). The phagocytic index of stimulated animals showed a significant increase from control (P < 0.001) and sham (P < 0.001). In addition, the stimulated animals showed a significant (P < 0.001) decrease in plasma corticosterone level when compared to sham and controls.

Observation-execution matching system for speech: a magnetic stimulation study.

Observation of limb movements in human subjects resulted in increased motor-evoked potential (MEP) amplitude elicited by magnetic stimulation of motor cortex in the muscles involved in that movement, suggesting that an observation-execution matching (OEM) system exists in humans. We investigated whether the OEM system is activated by speech gestures presented in the visual and auditory modalities. We found that visual observation of speech movement enhanced MEP amplitude specifically in muscles involved in production of the observed speech. In contrast, listening to the sound did not produce MEP enhancement. The findings suggest that the OEM system may be modality specific. It may be involved in action recognition in the visual modality, but is not responsible for perception of simple items of sound.

Norepinephrine and epinephrine levels in the brain of alloxan diabetic rats.

Thirty days after induction of experimental diabetes the brain catecholamines namely, norepinephrine (NE) and epinephrine (E) were studied in discrete brain regions (striatum, hippocampus, hypothalamus, midbrain, pons and medulla, cerebellum and cerebral cortex) in control, alloxan-diabetic untreated and insulin-treated diabetic rats. E showed significant increase in striatum, hippocampus and hypothalamus, whereas NE was increased in hypothalamus, and decreased in pons and medulla significantly in untreated diabetic rats. These effects were not seen in the insulin-treated diabetic rats.

Behavioural assessment of functional recovery after spinal cord hemisection in the bonnet monkey (Macaca radiata).

In spinal cord research, current approaches to behavioural assessment often fail in defining the exact nature of motor deficits or in evaluating the return of motor behaviour from lost functions following spinal cord injury. In addition to the assessment of gross motor behaviour, it is often appropriate to use complex tests for locomotion to evaluate the masked deficits in the evaluation of functional recovery after spinal cord injury. We designed a series of sensitive quantitative tests for reflex responses and complex locomotor behaviour in the form of a combined behavioural score (CBS) to assess the recovery of function in the Bonnet monkey (Macaca radiata). Monkeys were tested for various motor/reflex components, trained to cross different complex runways, and to walk on a treadmill bipedally. The overall performance of animal's motor behaviour and the functional status of individual limb movement during bipedal locomotion was graded and scored by the CBS. Surgical hemisection was then performed on the right side of the spinal cord at the T12-L1 level. Spinal cord hemisected animals showed a significant alteration in certain reflex responses such as grasping, extension withdrawal, and placing reflexes, which persisted through 1 year of follow-up. The spinal cord hemisected animals traversed the complex locomotor runways (Narrow beam and Grid runway) with more steps and few errors, at similar levels to control animals. These observations indicate that the various motor/reflex components and bipedal locomotor behaviour of spinal cord hemisected monkeys return to control levels gradually. These results are similar to those obtained in rat models by other investigators. These results demonstrate that the basic motor strategy and the spinal pattern generator for locomotion (SPGL) in adult monkeys for the accomplishment of complex motor tasks is similar, but not identical, to that in adult rats. This suggests that the mechanisms underlying recovery are probably similar in rats and monkeys, but that primates may take a longer duration to achieve the same functional end point.

In vitro effect of methanol on folate-deficient rat hepatocytes.

Methanol is primarily metabolized by oxidation to formaldehyde and then to formic acid. These processes are accompanied by formation of superoxide anion and hydrogen peroxide. This paper reports the in vitro antioxidant effect of vitamin E on isolated hepatocytes of folic acid deficient rats rendered so as to emulate a human hepatocyte model. These hepatocytes were treated with 320 microM of methanol per million cells and incubated for 30 min. The microsomal fraction of these hepatocytes showed a decreased level of superoxide dismutase (SOD), with increase in lipid peroxidation (LPO) shown by increase in recorded levels of malondialdehyde (MDA). Catalase activity was shown to be increased. Levels of reduced glutathione (GSH) were decreased and the activity of glutathione peroxidase (GSH-Px) and of glutathione reductase (GSSG-R) were not altered. The hepatocytes of folate deficient rats pretreated with vitamin E, when subjected to methanol treatment, showed no significant change in SOD levels and a significant decrease in MDA levels. The catalase activity in this group of animals showed a highly significant decrease. These animals had normal levels of GSH, while a significant fall in GSH-Px and GSSG-R levels were observed. These results suggest that Vitamin E exerts a protective effect on hepatocytes by acting as a free radical scavenger, proving its usefulness in treating methanol toxicity.

Effect of cassava consumption on open-field behavior and brain neurotransmitters in albino rats.

Diet exerts a critical influence on human biology and thus studies on the interrelationship of nutrition and behavior continues to be a major and important focus of research in the natural experimental sciences. Cassava is known to cause metabolic and neurological derangement on long-term consumption as a staple diet in the tropics. In this article we present the effects of cassava consumption on open-field behavior and catecholamine levels in the hypothalamus of albino rats. Cassava consumption for 30 days alters the emotional status of the rats, with changes in the basal neurotransmitter levels in the hypothalamus. The role of the cyanide (liberated from cassava) and protein deficiency (associated with cassava consumption) has been discussed.

Neurochemical and behavioural correlates in cassava-induced neurotoxicity in rats.

Chronic cyanide intoxication from cassava has been implicated as the cause for a degenerative neuropathy known widely as tropical ataxic neuropathy. An attempt has been made in this study to identify the specific cause for neuropathy caused by cassava using Wistar strain albino rats as the experimental animal model. The results of cassava fed animals were compared with control animals, animals given cyanide, malnourished animals and malnourished animals fed cyanide, to identify the causative factors. This study revealed that though the behavioural pattern in motor coordination of the cassava fed animals was similar to the other groups studied, the neurochemical basis for the observed behavioural pattern was unique for cassava. Hence the neurotoxicity of cassava could be attributed to unmetabolized linamarin, more than its nutritional status and/or cyanide toxicity.

An experimental analysis of the catecholamines in hyperglycemia and acidosis induced rat brain.

Hyperglycemia and acidosis are the hallmark of diabetes. Since these factors play an important role in the diabetic complications, we have studied the brain catecholamine levels in hyperglycemic and acidotic conditions per se. Experimentally induced hyperglycemia and acidosis are accompanied by significant alterations in the catecholamine levels in discrete areas of the brain. We and others have shown that chronic or acute diabetes in animals as well as in human results in altered neurotransmitter levels. In the present study, hyperglycemia maintained by daily external administration of glucose for thirty days showed increased level of dopamine in striatum and hippocampus, elevation of norepinephrine in hippocampus, and increased level of epinephrine in hypothalamus, midbrain and pons medulla. The ammonium chloride induced acidosis demonstrated significant elevation of dopamine in midbrain and significant increase of norepinephrine in hypothalamus and midbrain, and increased level of epinephrine in hypothalamus, pons medulla and cerebral cortex. On the other hand, sodium acetoacetate induced acidosis did not show any significant change in the level of catecholamines in any of the areas studied. In conclusion, the changes in catecholamine levels observed in experimentally induced hyperglycemic as well as in acidotic conditions are closely related to the changes observed in spontaneous or alloxan or streptozotocin diabetic animals, thereby suggesting that these conditions may be responsible for the changes observed in diabetic animals.

T lymphocyte subsets and immunoglobulins in intracranial tumor patients before and after treatment, and based on histological type of tumors.

It has been reported that nervous system and peripheral immune system communicate with each other and the peripheral immune status is depressed in some intracranial tumor (ICT) patients pre operatively. Little is known about the immune status of intracranial tumor patients during the post operative survival period. We thus investigated total T cells (CD 11+), helper/inducer (CD4+) T cells, suppressor/cytotoxic (CD8+) T cells, B cells (CD19+) and serum immunoglobulins in peripheral blood in certain ICT patients before and after treatment, and based on the histological type of the tumors. Post treatment analysis were conducted 30 days after surgical removal of tumor tissue in benign brain tumor patients and 30 days after chemo therapy (CT)/radiotherapy (RT) following surgical removal of tumor tissue in malignant brain tumor patients. Decreased CD11+, CD4+ and increased CD8+ T cell counts were observed in both benign and malignant tumor cases before treatment compared with control subjects. After treatment, CD4+ T cell count increased and CD8+ T cell count decreased than their pre treatment levels. Serum IgA and IgG levels were decreased in both benign and malignant brain tumor patients before treatment than in control subjects. Serum IgM level has been increased in both benign and malignant tumor patients before and after treatment than in control subjects. Anaplastic malignant astrocytoma, medulloblastoma and glioblastoma multiforme patients showed higher IgM level than astrocytoma, meningioma and ependymoma patients. In conclusions, the depressed host cellular immunity in benign and malignant tumor patients before treatment may be due to the changes in CD4+ and CD8+ counts in addition to tumour specific immunosuppressive factors. Treatment procedures such as surgery, CT and RT may play certain role in the post operative depressed immunosuppression in malignant tumor patients. Humoral immune mechanism (CD19+) in the ICT patients was less markedly affected.

Effect of chronic cyanide intoxication on memory in albino rats.

Cyanide is a chemical widely used in industry, and is a major environmental pollutant. Its toxicity is caused by inhibition of cytochrome oxidase resulting in histotoxic hypoxia. The effect of sublethal doses of cyanide on memory and hippocampal neurotransmitters was studied in male Wistar strain albino rats. Cyanide reduced the memory along with reduction in the levels of dopamine and 5-hydroxytryptamine in the hippocampus. Pre-existing malnutrition in the animals exaggerated these effects.

Effect of chronic protein restriction on motor co-ordination and brain neurotransmitters in albino rats.

In this study we evaluated the motor co-ordination in Wistar strain albino rats that were maintained on a protein-restricted diet for a period of 1 year immediately after the weaning period, by substituting 75% of the normal diet with a carbohydrate-rich diet deficient in protein, for a period of 1 year immediately after the weaning period. This type of chronic protein restriction caused disturbances in motor co-ordination. It also caused a significant reduction in the basal levels of dopamine, norepinephrine, epinephrine and serotonin along with their metabolites homovanillic acid (HVA), vanillyl mandelic acid (VMA) and 5-hydroxy indoleacetic acid (5HIAA) and precursor L-dopa in the corpus striatum and cerebellum. Changes in these neurotransmitters could have caused altered co-ordination in the protein-restricted animals.

Protective effect of diltiazem on cyanide-induced neurotoxicity in Wistar strain rats.

Cyanide is a well-established poison known for its rapid lethal action and toxicity. The central nervous system is one of the main target sites for cyanide toxicity. Cyanide not only alters brain biogenic amine levels but also the intracellular calcium levels in the neuronal cells. In the present study the role of calcium channel blocker diltiazem (DIL) in cyanide induced biogenic amine changes was evaluated in the Wistar strain rats. The protective effect of diltiazem pretreatment and diltiazem treatment along with cyanide on the dopaminergic system and the serotonergic system in the corpus striatum were studied. Diltiazem pretreatment was found to prevent cyanide induced changes in both the amines in the corpus striatum. These results suggest that diltiazem may mitigate the harmful effects of cyanide by interfering with influx of calcium ions and release of the biogenic amines.

Calcium ions: its role in cyanide neurotoxicity.

Cyanide is a well-established poison known for its rapid lethal action and toxicity. The central nervous system is one of the main target sites for cyanide toxicity. Cyanide also alters the brain biogenic amine levels. In the present study the role of calcium ions in cyanide toxicity was evaluated using the calcium channel blocker diltaizem (DIL) in Wistar strain albino rats. This study showed that DIL pretreatment prevented cyanide induced changes in the dopaminergic and serotonergic system in the corpus striatum.

Effect of Cassava on motor co-ordination and neurotransmitter level in the albino rat.

The root of Cassava, a tropical plant, is consumed in the tropics and has been attributed as the cause for various tropical neuropathies. This study aims to discover the neurotoxic effects of chronic cassava consumption of Indian origin and the effect of malnutrition. The assessment is based on the motor co-ordination and brain neurotransmitters in rats. Cassava consumption reduced the motor co-ordination, but the changes in neurotransmitter levels due to cassava consumption (except for 5HT in corpus striatum) was identical with malnutrition-induced changes, indicating that the toxicity of chronic cassava consumption (of Indian origin) is mainly due to the associated protein calorie malnutrition (PCM).