Membrane function alterations in erythrocytes from mood disorder patients

Objectives: To examine erythrocyte membrane functions in mood disorder patients and to establish possible diagnostic marker parameter(s).Design: Collection of blood samples from mood disorder patients and age-matched control volunteers.Preparation of erythrocyte membranes for the proposed studies.Setting: Out patients / in patients, psychiatry ward, Civil Hospital, Ahmedabad, Gujarat, India, Department of Biochemistry, Faculty of Science, M.S.University of Baroda, Vadodara, Gujarat, India.Subjects: Unipolar and bipolar subjects. Control subjects (randomly selected volunteers).Results: The most significant results were a duration dependent decrease in the TPL/CHL ratio (mole:mole),changes in both the substrate and temperature kinetics properties of AChE and elevated plasma BChE activity in the mood disorder patients.Conclusion: The results suggest that the altered lipid profiles and the TPL/CHL (mole: mole) ratio and the altered temperature-dependent activity coefficients of erythrocyte membrane AChE and elevated plasma BChE activities could serve as useful diagnostic pointers for mood disorders.Keywords:Membrane function; Erythrocytes; Mood disorderSA Psych Rev 2003;6:11-2

Study of protein synthesis in rat liver mitochondria: use of cycloheximide

1. Effects of short-term and long-term administration of cycloheximide on rat liver mitochondrial protein synthesis have been examined and were found to be different. 2. Long-term administration of cycloheximide resulted in inhibition of total cellular protein synthesis including that of mitochondria while, at short-term intervals, 8–10% of mitochondrial protein synthesis was cycloheximide-resistant. 3. The inhibitory effect was also reflected in terms of protein synthesizing ability of mitochondria in vitro, the inhibition becoming apparent at 40 min and showing progressive increase with time. 4. The observed inhibition of mitochondrial protein synthesis by cycloheximide was not due to either inhibition of energy metabolism or alteration of amino-acid pool. 5. Cycloheximide did not enter mitochondria or sonic preparation under conditions in vitro. On the other hand, after administration of [3H]cycloheximide, significant quantities of the label were found to be associated with mitochondria and mitoribosomes. 6. These results indicated that cycloheximide reached the site of action in mitochondria under conditions in vivo but was unable to do so in vitro. 7. The results are discussed to elucidate the possible mechanisms involved in the inhibition of truly mitochondrial protein synthesis by cycloheximide

Effect of long-term aluminum feeding on lipid/phospholipid profiles of rat brain myelin

Effect of long-term (90–100 days) exposure of rats to soluble salt of aluminum (AlCl(3)) on myelin lipid profile was examined. The long-term exposure to AlCl(3 )resulted in a 60 % decrease in the total phospholipid (TPL) content while the cholesterol (CHL) content increased by 55 %. Consequently the TPL / CHL molar ratio decreased significantly by 62 %. The phospholipid composition of the myelin membrane changed drastically; the proportion of practically all the phospholipid classes decreased by 32 to 60 % except for phosphatidylcholine (PC) and phosphatidylethanolamine (PE). Of the latter two, proportion of PC was unchanged while PE increased in proportion by 47 %. Quantitatively, all phospholipid classes decreased by from 42 to 76% with no change in the PE content. However the membrane fluidity was not altered in Al-treated rats. Many of the changes we observe here show striking similarities with the reported phospholipid profiles of Alzheimer brains

Compositional alterations in erythrocyte membranes in Type II diabetes

671-679Loss of erythrocyte membrane deformability is one of the most crucial factors in developing complications associated with Type II diabetes. The observed loss of erythrocyte membrane deformability could be related to structural changes in the membrane. In this context, here, we have made an attempt at gaining a better insight (quantitative as well as qualitative) into the protein and lipid contents in erythrocyte membranes and their interrelationships in Type II diabetes. Age matched control (n=12) and Type II diabetic subjects (n=22) were selected for this study. Morphological characteristics were studied by atomic force microscopy (AFM). AFM study confirmed remarkable alterations in morphology of the diabetic erythrocytes. In diabetic erythrocytes following changes were noted: (i) Significant increase in membrane as well as cytosolic proteins with a marginal increase in phospholipids content; (ii) The membrane total lipids:protein, phospholipids:protein, cholesterol:protein and phospholipids:cholesterol (mole:mole) ratios decreased significantly; (iii). A reproducible decrease in docosahexaenoic acid (DHA) and Omega-3 index with increase in Omega-6:Omega-3 ratio in membrane fatty acids; and (iv) The SDS-PAGE analysis indicated that all membrane proteins increased in almost equal proportion leading to increased membrane protein content. The observed compositional and stochiometric changes in lipids, proteins and their ratios may underlie morphological alterations and loss of deformability

Compositional alterations in erythrocyte membranes in Type II diabetes

Loss of erythrocyte membrane deformability is one of the most crucial factors in developing complications associated with Type II diabetes. The observed loss of erythrocyte membrane deformability could be related to structural changes in the membrane. In this context, here, we have made an attempt at gaining a better insight (quantitative as well as qualitative) into the protein and lipid contents in erythrocyte membranes and their interrelationships in Type II diabetes. Age matched control (n=12) and Type II diabetic subjects (n=22) were selected for this study. Morphological characteristics were studied by atomic force microscopy (AFM). AFM study confirmed remarkable alterations in morphology of the diabetic erythrocytes. In diabetic erythrocytes following changes were noted: (i) Significant increase in membrane as well as cytosolic proteins with a marginal increase in phospholipids content; (ii) The membrane total lipids:protein, phospholipids:protein, cholesterol:protein and phospholipids:cholesterol (mole:mole) ratios decreased significantly; (iii). A reproducible decrease in docosahexaenoic acid (DHA) and Omega-3 index with increase in Omega-6:Omega-3 ratio in membrane fatty acids; and (iv) The SDS-PAGE analysis indicated that all membrane proteins increased in almost equal proportion leading to increased membrane protein content. The observed compositional and stochiometric changes in lipids, proteins and their ratios may underlie morphological alterations and loss of deformability

A simplified fluorimetric method for corticosterone estimation in rat serum, tissues and mitochondria

48-53A simplified procedure has been developed for the extraction and estimation of corticosterone from rat serum, tissues and mitochondria. The suitably diluted samples were treated with freshly prepared chloroform: methanol mixture (2:1, v/v) and then extracted directly with the chloroform. Almost quantitative recoveries (90% and above) were obtained with the present method, compared to poor recoveries (65-81%) and variable results obtained by earlier procedure. Quantification of corticosterone content in tissues, such as liver, brain and heart, and in the mitochondria indicated significant concentration of corticosterone in tissues and mitochondria, as compared to the serum. The presence of corticosterone in the mitochondria suggests that the hormone may play a role in regulation of mitochondrial gene expression and/or their turnove

Kinetic attributes of rat liver microsomal adenosine 5’ triphosphate phosphohydrolase (ATPase)

252-259The kinetic properties of the rat liver microsomal ATPase, with respect to Na+, K+ and ATP requirements were examined. Presence of Na+ and K+ or both hardly caused any stimulation of the enzyme activity. The Km values for Na+ and K+were substantially low (0.32 and 0.05 mM, respectively), compared to those reported for the Na+, K+ ATPases from different tissues. Substrate kinetics studies revealed that in the absence of Na+ and K+, ATP is an activator of the enzyme. The enzyme displayed increased activity with increase in the energy of activation in the absence of Na+ and K+. The activity was partially inhibited by ouabain only in the presence of Na+ and K+. The results suggest that the liver microsomal enzyme is not a Na+, K+ ATPase, but has requirement of monovalent cations for the regulation of its activity. Also, the β3 subunit of the enzyme has a Km lowering effect

Kinetic Attributes of Na +

Kinetic properties of Na , K ATPase of membranes from rat and human erythrocytes were examined. The enzyme stability decreased with incubation time. The V of the human enzyme was about 4 times lower than the values of the rat enzyme. However the energies of activation were higher. Phase transition temperature for the rat and the human enzyme was 24 °C and 17 °C, respectively. The human erythrocyte membranes were characterized by lower total phospholipid and cholesterol contents and were relatively more fluid. The human membranes contained lower proportions of acidic phospholipids which correlated well with the lower V of the enzyme; the proportion of lysophosphoglyceride and sphingomyelin was higher in the human membrane

Paracetamol hepatotoxicity and microsomal function

The effect of paracetamol-induced hepatotoxicity in rats (650 mg/kg) on microsomal function was examined. Paracetamol treatment resulted in lowered Na +,K +-ATPase activity in the microsomes with decrease in V max of the low affinity high V max component II. However, the temperature kinetics was not influenced significantly. The total phospholipid and cholesterol contents as well as lipid peroxidation in the microsomes were unchanged. However, content of acidic phospholipids: phosphatidylserine and phosphatidylinositol decreased by 50% with a reciprocal increase in the sphingomyelin content; the lysophosphoglyceride content increased by 12-fold. The microsomal membrane appeared to be more fluidized following paracetamol treatment. Paracetamol treatment also resulted in a significant reduction in the sulfhydryl groups content