Comparison of haematological and biochemical changes between non-anaemic and anaemic primigravid women in a north Indian population to establish normative values.

Pregnancy is accompanied by several haemodynamic, biochemical and haematological changes, which may lead to severe problems, if they are not suitably addressed. The current study highlights the haematological and biochemical differences observed in anaemic (AP) and non-anaemic primigravida (NAP), in the 2nd trimester, in a north Indian population. There were significant differences (p < 0.05) in the body weight and body temperature of NAP compared with AP. A significant decrease (p < 0.001) in haematological parameters including haemoglobin, haematocrit, erythrocyte count, MCH and MCHC, was observed in AP; however, MCV was found to be significantly higher (p = 0.038). Many biochemical parameters viz. potassium, albumin, total protein and calcium levels were significantly reduced (p < 0.01) in AP, except alkaline phosphatase whose level was found significantly increased (p < 0.01). The findings of the study suggest that haematological and biochemical changes take place in anaemia during pregnancy. Further, the results obtained should be used for establishing normative values for similar populations.

Increased oxidative stress is associated with the development of organophosphate-induced delayed neuropathy.

Organophosphate-induced delayed neuropathy (OPIDN) is a progressive neuropathic disorder that manifests in days to weeks following exposure to an acute dose of organophosphates. The precise mechanism involved in the development of OPIDN is not clear as it develops after many days of the cessation of cholinergic crisis. The present study has been designed to understand the role of oxidative stress in the development of OPIDN, wherein neuropathy was developed by the administration of acute dose of monocrotophos (MCP) or dichlorvos (2,2-dichlorovinyl dimethyl phosphate (DDVP)) to rats. Significant motor deficits in terms of reduced spontaneous locomotor activity and performance on narrow beam test were observed after 14 days of exposure to MCP or DDVP, which persisted even on day 28, suggesting the development of OPIDN. Rats with OPIDN also exhibited an increase in malondialdehyde levels along with a decrease in thiol content in cerebral cortex, cerebellum and brain stem. Concomitantly, the activities of antioxidant enzymes, superoxide dismutase, catalase, glutathione peroxidase and glutathione reductase were reduced in the three brain regions. The biochemical and functional changes were associated with histological alterations in the brain regions studied. The results clearly indicate that the development of OPIDN is mediated in part through an increased oxidative stress and suggest that the strategies aimed at restoration of antioxidant capacity may be beneficial for the individuals with OPIDN-like symptoms.

Alterations in Ca²âº homeostasis and oxidative damage induced by ethion in erythrocytes of Wistar rats: ameliorative effect of vitamin E.

Organophosphate (OP) insecticides have been reported to induce oxidative stress due to lipid peroxidation and alteration in defense mechanisms. It is known that calcium content in erythrocytes plays a very important in normal physiology of cells. Erythrocytes are a very convenient model to understand the susceptibility of membrane to oxidative damage induced by various xenobiotic compounds. The aim of present study was to investigate the effects of ethion induced oxidative damage, alterations in membrane bound enzymes and Ca(2+) homeostasis and a possible protective role of vitamin E. Adult male albino rats of Wistar strain were orally administered ethion and vitamin E daily for 28 days. Animals were randomly divided into four groups: control; ethion treated (2.7 mg/kgbw/day); vitamin E treated (50mg/kg of bw/day); ethion+vitamin E treated. The animals were sacrificed after 7, 14, 21 and 28 days. Erythrocyte membranes were prepared and analyzed for protein, lipid peroxidation (LPO) and membrane bound ATPases. Furthermore, Ca(2+) homeostasis as function of time and concentration was evaluated in erythrocytes. The results from the present study show that in vivo administration of ethion resulted in oxidative damage to erythrocyte membranes as evident by increased lipid peroxidation. The increased LPO following ethion intoxication was accompanied by significant decrease in the activities of Na(+)/K(+)-ATPase, Mg(2+)-ATPase and Ca(2+)-ATPase and disturbed Ca(2+)homeostasis in erythrocytes. Furthermore, vitamin E treatment had a beneficial effect by decreasing lipid peroxidation; partially restoring activities of membrane bound ATPases and Ca(2+) homeostasis. The present study suggests that ethion exerts its toxic effect by increasing LPO, altering the activity of membrane bound enzymes and disturbing Ca(2+) homeostasis. Vitamin E treatment ameliorated the toxic effects of ethion suggesting its role as a potential antioxidant.

Biochemical and morphological perturbations in rat erythrocytes exposed to ethion: protective effect of vitamin E.

Erythrocyte membranes are an excellent model system to study interaction of pro-oxidants with membranes. The aim of the present study is to examine the effect of vitamin E on ethion-induced biochemical and morphological alterations in erythrocytes. Ethion was administered to the rats orally at a daily dose of 2.7 mg/kg body weight for a period of 7, 14, 21 and 28 days. The results from the present study show that administration of ethion resulted in oxidative damage to erythrocyte membranes as evident by increased lipid peroxidation and decreased phospholipid content. This was accompanied by decrease in membrane cholesterol levels. In addition, ethion exposure inhibited the activities of membrane bound enzymes; Na+ K+ ATPase and Mg2+ATPase. Scanning electron micrographs of erythrocytes from animals exposed to ethion revealed morphological changes. Supplementation of vitamin E (50 mg/kg body weight) to ethion exposed animals ameliorated the ethion-induced oxidative stress, restored membrane lipid composition and activity of membrane bound enzymes along with erythrocyte shape. The results clearly demonstrate that ethion-induced damage involves increase in oxidative stress that results in alterations in erythrocyte membrane structure and function. Furthermore, supplementation with vitamin E reversed ethion induced alterations suggesting its beneficial role in individuals exposed to ethion.

Effects of Azadirachta indica on certain hematological parameters during benzo(a)pyrene induced murine forestomach tumorigenesis.

OBJECTIVES: Exposure to environmental toxicants/carcinogens, the process of carcinogenesis itself and cancer treatments lead to several secondary pathologies including hematological complications. Considering versatile pharmacological potentials of Azadirachta indica (A. indica), the present study was designed to evaluate its effects on certain hematological parameters in benzo(a)pyrene [B(a)P] induced murine forestomach tumorigenesis bioassay protocol. METHODS AND RESULTS: For tumor induction, starting from 3rd week of experimentation, female Balb/c mice received B(a)P intragastric instillations (40 mg/kg bwt) twice a week for 4 weeks. Aqueous A. indica leaf extract (AAILE) was orally administered (100 mg/kg bwt) using blunt-tipped canula on alternate days throughout experimentation. The study was continued for 24 weeks and certain hematological parameters were examined at 4 week intervals. In mice receiving only B(a)P, hemoglobin (Hb), red blood cells (RBCs), white blood cells (WBCs), lymphocytes and monocytes decreased whereas neutrophils increased when compared to controls. However, A. indica reversed these alterations as seen in mice that received AAILE along with B(a)P when compared to only B(a)P receiving mice. In only AAILE receiving mice, increased Hb, RBCs, WBCs, lymphocytes and monocytes with decreased neutrophils were observed in comparison to control. Also, changes in eosinophils and basophils upon B(a)P exposure and their modulation by AAILE was observed. CONCLUSIONS: These findings strongly suggested the favorable effects of A. indica on hematological parameters studied and their significance with respect to overall well being, process of tumorigenesis and its chemoprevention have been discussed in the current research manuscript.

Hyperglycemia-induced alterations in synaptosomal membrane fluidity and activity of membrane bound enzymes: beneficial effect of N-acetylcysteine supplementation.

Diabetic encephalopathy is characterized by impaired cognitive functions that appear to underlie neuronal damage triggered by glucose driven oxidative stress. Hyperglycemia-induced oxidative stress in diabetic brain may initiate structural and functional changes in synaptosomal membranes. The objective of the present study was to examine the neuroprotective role of N-acetylcysteine (NAC) in hyperglycemia-induced alterations in lipid composition and activity of membrane bound enzymes (Na(+),K(+)-ATPase and Ca(2+)-ATPase) in the rodent model of type 1 diabetes. Male Wistar rats weighing between 180 and 200 g were rendered diabetic by a single injection of streptozotocin (50 mg/kg body weight, i.p.). The diabetic animals were administered NAC (1.4-1.5 g/kg body weight) for eight weeks and lipid composition along with membrane fluidity were determined. A significant increase in lipid peroxidation was observed in cerebral cortex of diabetic rats. NAC administration on the other hand lowered the hyperglycemia-induced lipid peroxidation to near control levels. The increased lipid peroxidation following chronic hyperglycemia was accompanied by a significant increase in the total lipids which can be attributed to increase in the levels of cholesterol, triglycerides and glycolipids. On the contrary phospholipid and ganglioside levels were decreased. Hyperglycemia-induced increase in cholesterol to phospholipid ratio reflected decrease in membrane fluidity. Fluorescence polarization (p) with DPH also confirmed decrease in synaptosomal membrane fluidity that influenced the activity of membrane bound enzymes. An inverse correlation was found between fluorescence polarization with the activities of Na(+),K(+)-ATPase (r(2)=0.416, P<0.05) and Ca(2+) ATPase (r(2)=0.604, P<0.05). NAC was found to significantly improve lipid composition, restore membrane fluidity and activity of membrane bound enzymes. Our results clearly suggest perturbations in lipid composition and membrane fluidity as a major factor in the development of diabetic encephalopathy. Furthermore, NAC administration ameliorated the effect of hyperglycemia on oxidative stress and alterations in lipid composition thereby restoring membrane fluidity and activity of membrane bound enzymes.

Role of the oestrogen receptors GPR30 and ERalpha in peripheral sensitization: relevance to trigeminal pain disorders in women.

Oestrogen increases facial allodynia through its actions on activation of the MAPK extracellular-signal regulated kinase (ERK) in trigeminal ganglion neurons. This goal of study was to determine which oestrogen receptor is required for behavioural sensitization. Immunohistochemical studies demonstrated the presence of oestrogen receptor alpha (ERalpha) in nuclei of larger neurons and cytoplasm of smaller neurons, and the novel oestrogen receptor G-protein coupled receptor 30 (GPR30) in small diameter neurons that also contained peripherin, a marker of unmyelinated C-fibres. Specific agonists for ERalpha (PPT) and GPR30 (G-1), but not ERbeta (DPN), activated ERK in trigeminal ganglion neurons in vitro. Both G-1 and PPT treatment increased allodynia after CFA injections into the masseter of ovariectomized Sprague-Dawley rats. Treatment with oestrogen increased expression of ERalpha but not GPR30, while masseter inflammation increased GRP30 but not ERalpha. Differential modulation of these ERK-coupled receptors by oestrogen and inflammation may play a role in painful episodes of temporomandibular disorder and migraine.

Oestrogen increases nociception through ERK activation in the trigeminal ganglion: evidence for a peripheral mechanism of allodynia.

The mitogen-activated protein kinase, extracellular signal-regulated kinase (ERK), is activated in experimental models of chronic pain, and is also activated by oestrogen. We used an established model of inflammatory trigeminal pain, injection of Complete Freund's Adjuvant (CFA) into the masseter muscle, to determine whether ERK activation may play a role in hormone-related trigeminal pain disorders. We measured withdrawal responses to stimulation of the masseter (V3, primary allodynia) and whisker pad (V2, secondary allodynia) using graded monofilaments. Oestrogen treatment in the presence of inflammation increased withdrawal response to stimulation of both masseter and whisker pad compared with inflammation alone, indicating an additive effect of inflammation and oestrogen on both primary and secondary allodynia. We examined ERK activation in trigeminal ganglia from each treatment group using western blot and immunohistochemistry. Both masseter inflammation and oestrogen treatment increased ERK activation, and combined treatment had an additive effect. Both masseter inflammation and oestrogen increased the percentage of pERK immunoreactive neurons in divisions 1 and 2 (V1/2), and combined treatment increased pERK immunoreactivity in V1/2 compared with inflammation alone. We stereotactically administered ERK antagonist U0126, or inactive control U0124, to the trigeminal ganglion of CFA+E2-treated rats. U0126 decreased withdrawal responses to mechanical stimulation of the whisker pad compared with U0124-treated rats. Because the secondary allodynia in V2 after inflammation in V3 was reduced by antagonizing ERK activation in the periphery, these data suggest a peripheral component to secondary allodynia mediated through ERK activation.

Marjolin's ulcer of the scalp invading calvarial bone, dura and brain.

A Marjolin's ulcer arising in a chronic scalp scar is reported. Malignant transformation occurred after four decades. The tumour had invaded the skull, dura mater and underlying brain tissue. This case illustrates the importance of appropriate and expeditious initial wound management of scar ulcers with high malignant potential to prevent this devastating outcome and also the role of reconstructive surgical techniques in radical surgery for scalp cancers.

Identification of the pathway of alpha-oxidation of cerebronic acid in peroxisomes.

Cerebronic acid (2-hydroxytetracosanoic acid), an alpha-hydroxy very long-chain fatty acid (VLCFA) and a component of cerebrosides and sulfatides, is unique to nervous tissues. Studies were carried out to identify the pathway and the subcellular site involved in the oxidation of cerebronic acid. The results from these studies revealed that cerebronic acid was catabolized by alpha-oxidation to CO2 and tricosanoic acid (23:0). Studies with subcellular fractions indicated that cerebronic acid was alpha-oxidized in fractions having particulate bound catalase and enzyme systems for the beta-oxidation of VLCFA (e.g., lignoceric acid), suggesting peroxisomes as the subcellular organelle responsible for alpha-oxidation of cerebronic acid. Etomoxir, an inhibitor of mitochondrial fatty acid oxidation, had no effect on cerebronic acid alpha-oxidation. Further, cerebronic acid oxidation was found to be dependent on the presence of NAD+ but not FAD, NADPH, ATP, Mg2+, or CoASH. Intraorganellar localization studies indicated that the enzyme system for the alpha-oxidation of cerebronic acid was associated with the peroxisomal limiting membranes. Studies on cultured fibroblasts from normal subjects and patients with peroxisomal disorders indicated an impairment of alpha-oxidation of cerebronic acid in cell lines that lack peroxisomes [e.g., Zellweger syndrome (ZS)]. On the other hand, alpha-oxidation of cerebronic acid was found to be normal in cell lines from X-linked adrenoleukodystrophy, adult Refsum disease, and rhizomelic chondrodysplasia punctata. Our results clearly demonstrate that alpha-oxidation of alpha-hydroxy VLCFA (cerebronic acid) is a peroxisomal function and that this oxidation is impaired in ZS. Furthermore, this alpha-oxidation enzyme system is distinct from the one for the alpha-oxidation of beta-carbon branched-chain fatty acids (e.g., phytanic acid).

Increased peroxisomal fatty acid beta-oxidation and enhanced expression of peroxisome proliferator-activated receptor-alpha in diabetic rat liver.

To determine whether the increased fatty acid beta-oxidation in the peroxisomes of diabetic rat liver is mediated by a common peroxisome proliferation mechanism, we measured the activation of long-chain (LC) and very long chain (VLC) fatty acids catalyzed by palmitoyl CoA ligase (PAL) and lignoceryl CoA ligase and oxidation of LC (palmitic acid) and VLC (lignoceric acid) fatty acids by isotopic methods. Immunoblot analysis of acyl-CoA oxidase (ACO), and Northern blot analysis of peroxisome proliferator-activated receptor (PPAR-alpha), ACO, and PAL were also performed. The PAL activity increased in peroxisomes and mitochondria from the liver of diabetic rats by 2.6-fold and 2.1 -fold, respectively. The lignoceroyl-CoA ligase activity increased by 2.6-fold in diabetic peroxisomes. Palmitic acid oxidation increased in the diabetic peroxisomes and mitochondria by 2.5-fold and 2.7-fold, respectively, while lignoceric acid oxidation increased by 2.0-fold in the peroxisomes. Immunoreactive ACO protein increased by 2-fold in the diabetic group. The mRNA levels for PPAR-alpha, ACO and PAL increased 2.9-, 2.8- and 1.6-fold, respectively, in the diabetic group. These results suggest that the increased supply of fatty acids to liver in diabetic state stimulates the expression of PPAR-alpha and its target genes responsible for the metabolism of fatty acids.

Hepatoprotective effects of Liv-52 on ethanol induced liver damage in rats.

The mechanism of protective effects of Liv-52, a multiherbal hepatoprotective drug, on ethanol induced hepatic damage has been investigated. The results indicate that Liv-52 treatment prevents ethanol induced increase in the activity of the enzyme gamma-glutamyl transpeptidase. Concomitantly there was also a decrease in ethanol accentuated lipid peroxidation in liver following Liv-52 treatment. The activity of antioxidant enzymes; superoxide dismutase, glutathione peroxidase and the levels of glutathione were decreased following ethanol ingestion. Liv-52 treatment was found to have protective effects on the activity of superoxide dismutase and the levels of glutathione. The results obtained from the study indicate hepatoprotective nature of Liv-52 which might be attributed to its ability to inhibit lipid peroxidation.

Localization of nervonic acid beta-oxidation in human and rodent peroxisomes: impaired oxidation in Zellweger syndrome and X-linked adrenoleukodystrophy.

Studies with purified subcellular organelles from rat liver indicate that nervonic acid (C24:1) is beta-oxidized preferentially in peroxisomes. Lack of effect by etomoxir, inhibitor of mitochondrial beta-oxidation, on beta-oxidation of lignoceric acid (C24:0), a peroxisomal function, and that of nervonic acid (24:1) compared to the inhibition of palmitic acid (16:0) oxidation, a mitochondrial function, supports the conclusion that nervonic acid is oxidized in peroxisomes. Moreover, the oxidation of nervonic and lignoceric acids was deficient in fibroblasts from patients with defects in peroxisomal beta-oxidation [Zellweger syndrome (ZS) and X-linked adrenoleukodystrophy (X-ALD)]. Similar to lignoceric acid, the activation and beta-oxidation of nervonic acid was deficient in peroxisomes isolated from X-ALD fibroblasts. Transfection of X-ALD fibroblasts with human cDNA encoding for ALDP (X-ALD gene product) restored the oxidation of both nervonic and lignoceric acids, demonstrating that the same molecular defect may be responsible for the abnormality in the oxidation of nervonic as well as lignoceric acid. Moreover, immunoprecipitation of activities for acyl-CoA ligase for both lignoceric acid and nervonic acid indicate that saturated and monoenoic very long chain (VLC) fatty acids may be activated by the same enzyme. These results clearly demonstrate that similar to saturated VLC fatty acids (e.g., lignoceric acid), VLC monounsaturated fatty acids (e.g., nervonic acid) are oxidized preferentially in peroxisomes and that this activity is impaired in X-ALD. In view of the fact that the oxidation of unsaturated VLC fatty acids is defective in X-ALD patients, the efficacy of dietary monoene therapy, "Lorenzo's oil," in X-ALD needs to be evaluated.

Altered cholinergic metabolism in rat CNS following aluminum exposure: implications on learning performance.

The effects of Al on the central cholinergic system have been studied. Al, at a dose of 10 mg/kg of body weight/day for 4 weeks, had a deleterious effect on the activities of biosynthetic (choline acetyltransferase) and hydrolytic (acetylcholinesterase) enzymes of the neurotransmitter acetylcholine. The levels of acetylcholine were also significantly lowered in different brain regions at the end of the dose regimen. There was a significant decrease in high-affinity choline uptake following Al exposure. Muscarinic acetylcholine receptor binding studies revealed a decreased number of binding sites (Bmax), with the maximum effects being manifested in the hippocampus. Exogenous addition of 10 microM desferrioxamine restored the muscarinic receptor binding completely. The impaired cholinergic functioning had severe effects on cognitive functions. Neurobehavioral deficits were manifested in terms of decreased active (52%) and passive (73.30%) avoidance tests. The results suggest that Al exerts its toxic effects by altering cholinergic transmission, which is ultimately reflected in neurobehavioral deficits.

Effect of lead on lipid peroxidation in liver of rats.

The present study was undertaken to understand the biochemical mechanisms of lead toxicity in liver. We observed a significant accumulation of lead in liver following lead treatment, resulting in accentuation of lipid peroxidation. Concomitant to the increase in lipid peroxidation, the activities of antioxidant enzymes, viz., superoxide dismutase, catalase, glutathione peroxidase, and glutathione reductase, were significantly inhibited. A decrease in reduced glutathione with a simultaneous increase in oxidized glutathione was observed following lead exposure, resulting in a reduced GSH/GSSG ratio. These results indicate that lead exerts its toxic effects by enhancing peroxidative damage to the membranes, thus compromising cellular functions.