Toxic copper level increases erythrocyte glycolytic rate, glutathione production and alters electrolyte balance in male Wistar rats.

BACKGROUND: Copper is a micronutrient vital to several cellular energy metabolic processes and drives erythropoiesis. However, it disrupts cellular biological activities and causes oxidative damage when in excess of cellular needs. This study investigated the effects of copper toxicity on erythrocyte energy metabolism in male Wistar rats. METHODS: Ten Wistar rats (150-170 g) were randomly divided into 2 groups: control (given 0.1 ml distilled water) and copper toxic (given 100 mg/kg copper sulphate). Rats were orally treated for 30 days. Blood, collected retro-orbitally after sodium thiopentone anaesthesia (50 mg/kg i.p.) into fluoride oxalate and EDTA bottles, was subjected to blood lactate assay and extraction of red blood cell respectively. Red blood cell nitric oxide (RBC NO), glutathione (RBC GSH), adenosine triphosphate (RBC ATP) levels, RBC hexokinase, glucose-6-phosphate (RBC G6P), glucose-6-phosphate dehydrogenase (RBC G6PDH), and lactate dehydrogenase (RBC LDH) activity was estimated spectrophotometrically. Values (Mean±SEM, n = 5) were compared by Student's unpaired T-test at p < 0.05. RESULTS AND CONCLUSION: Copper toxicity significantly increased RBC hexokinase (23.41 ± 2.80 µM), G6P (0.48 ± 0.03 µM), G6PDH (71.03 ± 4.76nmol/min/ml) activities, ATP (624.70 ± 57.36 µmol/gHb) and GSH (3.08 ± 0.37 µM) level compared to control (15.28 ± 1.37 µM, 0.35 ± 0.02 µM, 330.30 ± 49.58 µmol/gHb, 54.41 ± 3.01nmol/min/ml and 2.05 ± 0.14 µM respectively, p < 0.05). Also, RBC LDH activity (145.00 ± 19.88mU/ml), NO (3.45 ± 0.25 µM) and blood lactate (31.64 ± 0.91 mg/dl) level were lowered significantly compared to control (467.90 ± 94.23mU/ml, 4.48 ± 0.18 µM and 36.12 ± 1.06 mg/dl respectively). This study shows that copper toxicity increases erythrocyte glycolytic rate and glutathione production. This increase could be connected to a compensatory mechanism for cellular hypoxia and increased free radical generation.

Mitochondrial diversity of Yoruba and Fulani chickens: A biodiversity reservoir in Nigeria.

Poultry are the most widely distributed type of livestock in Nigeria. Indigenous chickens are extremely common throughout the country. Indeed, approximately 83 million chickens are raised in extensive systems and 60 million in semi-intensive systems. To provide the first comprehensive overview of the maternal lineages in Southwest Nigeria, we analyzed 96 mitochondrial DNA control region sequences from 2 indigenous chicken ecotypes: Fulani and Yoruba. All samples belonged to the most frequent haplogroup (E) in Africa and Europe and showed noticeably low haplotype diversity. Although only 11 different haplotypes were detected, with 2 of them never found before in Nigeria, the presence of unique sequences among our indigenous samples testified to their status as an important genetic resource to be preserved. Furthermore, a total of 7,868 published sequences were included in the comparative analysis, which revealed an east-west geographic pattern of haplogroup distribution and led to the conclusion that the gene flow from Southeastern Asia mainly involved one mitochondrial clade. Moreover, owing to the extensive genetic intermixing among Nigerian chickens, conservation efforts are required to safeguard the extant mitochondrial variability in these indigenous ecotypes and establish future improvement and selection programs.

Amelioration of thyroid dysfunction by magnesium in experimental diabetes may also prevent diabetes-induced renal impairment.

BACKGROUND: Diabetes mellitus has been reported to cause thyroid dysfunction, which may also impair renal function. Magnesium has been reported to exert ameliorative effects in diabetes mellitus. This study investigated thyroid and renal functions in experimental type-2-diabetic Wistar rats. METHODS: Experimental type-2-diabetes was induced using short duration high-fat (30%) diet feeding followed by single-dose streptozotocin (35 mg/kg i.p.). Fifty rats were randomly divided into five equal groups consisting of control, diabetes untreated, diabetes treated with either magnesium (250 mg/kg) or metformin (250 mg/kg) and diabetes treated with both metformin and magnesium simultaneously.All treatments were carried out orally for 14days post-diabetes induction. Body weight and blood glucose was monitored using the tail tipping method before diabetes induction and thereafter on days 1,7,14 post-treatment respectively. Thereafter, blood samples were collected by cardiac puncture after light anesthesia into plain and EDTA sample bottles. Total protein, albumin, globulin (plasma) and insulin (serum) were assayed in all samples obtained. Thyroid stimulating hormone (TSH), triiodothyronine, thyroxine was also evaluated (n = 5/group) in serum while blood urea nitrogen (BUN), creatinine was assessed (n = 5/group) in plasma. Kidney homogenates were obtained per group and analyzed for renal superoxide dismutase (SOD), reduced glutathione (GSH) and lipid peroxidation (MDA). Kidney histology was also evaluated per group using both Haematoxylin and Eosin and periodic acid Schiff stains. RESULTS: Body weight, blood glucose, insulin, renal MDA was increased in diabetic untreated compared to other groups. Reductions (P < 0.05) in TSH, triiodothynine, Renal SOD and GSH levels where observed in diabetic untreated compared to other groups. Renal histology in diabetic untreated showed glomerula sclerosis, fused messengial cells and either collapsed tubular lumen or lumen with eosinophilic renal cast. These pathologies where partially reversed in the other experimental groups. CONCLUSION: This study suggests that thyroid and renal impairment may be present in experimental type-2-diabetes. Treatment with oral magnesium may cause a partial restoration of thyroid function that may impede the development of renal dysfunction.

Pathophysiology of iron overload-induced renal injury and dysfunction: Roles of renal oxidative stress and systemic inflammatory mediators.

Iron-overload has been recognized as a risk factor for organ dysfunction and damage resulting in diseases such as liver and heart disease, diabetes mellitus, and neurodegenerative diseases. This study investigated renal function and some systemic inflammatory indices in iron-overloaded male Wistar rats. Thirty animals were equally distributed into 3groups and treated daily i.p. with either normal saline (0.2 ml; control), iron (as ferrous sulphate) (15 mg/kg) or iron (30 mg/kg) for 21days respectively. Post-treatment, blood samples were obtained from each animal by cardiac puncture after light anaesthesia into plain sample bottles. Iron, ferritin, transferrin, creatinine, urea, albumin, total protein, interleukin-6 (IL-6), prostaglandins-E2 and tumor necrosis factor-α (TNF-α) were analysed in serum. Kidney homogenates were obtained per group and analysed for superoxide dismutase (SOD), total antioxidant capacity (TAC), reduced glutathione (GSH), lipid peroxidation (MDA) and nitric oxide (NO). Kidney histology was evaluated per group using both Haematoxylin and Eosin and periodic acid Schiff stains. Iron-overload caused a graded increase (p < 0.05) in serum iron, ferritin, transferrin, creatinine, urea, IL-6, TNF-α, TAC, MDA and NO levels as well as a reduction in albumin levels, renal SOD and GSH in groups 2 (iron 15 mg/kg) and 3 (iron 30 mg/kg) respectively compared to control. Histological evaluation of the kidney showed structural and tubular aberrations consistent with renal damage via inflammatory processes in iron overloaded rats. Our present study suggests that iron-overloading causes renal dysfunction by triggering the evolution of several inflammatory mediators which lead to a cascade of systemic and renal inflammatory processes that alter renal structure and function.

Metformin Effects are Augmented by Chronic Intermittent Cold Stress in High Fat Diet Fed Male Wistar Rats.

This study investigated the effects of metformin on some glucose regulatory indices in high fat diet (HFD) fedmale Wistar rats exposed to room temperature and chronic intermittent cold stress (CICS). Thirty rats were randomly dividedinto 5 groups. Group 1(control) was maintained on standard rat chow while groups 2-5 were maintained on HFD for 8weeksrespectively prior to experimental procedures. Control, group 2(HFD untreated) and group 3(HFD+metformin (250mg/kg)were exposed to room temperature while groups 4(HFD untreated+CICS) and 5(HFD+CICS+metformin) were exposed toCICS for 21days. Blood glucose was monitored before initial exposure to HFD and on days 1,7,14 and 21 respectively.Blood samples (5mls) were thereafter collected by cardiac puncture following light ether anaesthesia, serum was obtainedand analysed for insulin, cortisol, and lipid profile using laboratory kits. Pancreatic ß-cell function and insulin resistancewere estimated using the Homeostasis Model Assessment equations. It was observed that blood glucose reduced significantlyin groups 2-4 on day21 compared to day1 values. At day 21 post-treatment, insulin level and insulin resistance were increasedwhile cholesterol levels were reduced in all HFD groups compared to control. Cortisol was increased in group 2 but reducedin groups 3-4 compared to control. HDL was reduced in groups 2-3 while liver glycogen was increased in groups 2, 3 and 5compared to control. Beta cell function and muscle glycogen were increased while LDL and triglyceride were reduced ingroups 2-4 compared to control. In conclusion, metformin ameliorates high-fat diet (HFD) induced impairment of glucoseand lipid regulatory indices by facilitating an increase in the storage of glycogen in the liver and muscle. Chronic intermittentcold stress exposure in HFD rats does not ameliorate insulin resistance but reduces impaired glucose and lipid regulatoryindices likely through an increase in adaptive thermo-genic mechanisms. The actions of metformin in reducing stressfulstimulus and preventing pre-diabetes syndrome in HFD fed rats are augmented by exposure to chronic intermittent coldstress.

Oral Magnesium Treatment Reduces Anemia and Levels of Inflammatory Markers in Experimental Diabetes.

Magnesium has been reported to improve glucose utilization in diabetes mellitus. However, information on its effects on anemic and inflammatory markers in diabetes mellitus is limited. This study investigated the effect of oral magnesium (Mg) treatment on some markers of anemia and inflammation in 25 male Wistar rats. Rats (200 ± 15 g) were randomly divided into five groups (n = 5). Group 1 was control (received orally 0.2 mL distilled water daily), group 2 (Diabetic Untreated), group 3 (Diabetic Mg treated-100 mg/kg bw), group 4 (Diabetic Mg treated-250 mg/kg bw), group 5 (Diabetic Insulin treated-1 IU/kg bw). Diabetes was induced with a single dose of alloxan (100 mg/kg intraperitoneal (i.p.)). All treatments were done for 14 days. Anemic and inflammatory markers were investigated on blood samples obtained from each animal using standard laboratory methods. Significant increase (p < 0.05) in total white blood cell (WBC) count was observed in diabetic untreated rats (7.67 ± 0.397 × 109/L) compared to control (5.88 ± 0.25 × 109/L), DMg 100 (5.86 ± 0.74 × 109/L) and DMg 250 (5.06 ± 0.78 × 109/L). Hemoglobin concentration, packed cell volume (PCV) and red blood cell (RBC) count was decreased (p < 0.05) in DU compared to control, DMg 100, and DI rats. Erythrocyte sedimentation rate (ESR) was significantly increased (p < 0.05) in DU compared to control, DMg 100, DMg 250, and DI groups. Fibrinogen level was increased (p < 0.05) in DU rats (0.44 ± 0.02 g/dL) compared to control(0.26 ± 0.02 g/dL). Values obtained in DMg 100 (0.30 ± 0.03 g/dL), DMg 250 (0.22 ± 0.04 g/dL), and DI (0.36 ± 0.02 g/dL) rats were comparable to control (0.26 ± 0.02 g/dL). Total protein, albumin, and globulin levels were decreased in DU rats compared to normal control, DMg 100, DMg 250, and DI rats. In conclusion, anemia and increased hematologic and metabolic inflammatory markers may be associated with untreated diabetes mellitus. Treatment of alloxan-induced diabetic rats with magnesium improved the anemic state and reduced hematologic and metabolic inflammatory markers.

Oral magnesium reduces gastric mucosa susceptibility to injury in experimental diabetes mellitus.

This study investigated the effect of magnesium on the gastric defence mechanism in alloxan-diabetic male Wistar rats. Sixty rats were randomly divided into 2 groups, A (n=40) and B (n=20). Each group was subdivided into control, diabetic untreated (DU), diabetic magnesium (250mg/kg) treated (DMg250) and diabetic insulin (3IU/kgs.c) treated (DI). Diabetes was induced with alloxan (120mg/kg) and both groups were treated for 14days. By day 14, group A rats were sacrificed, the stomach excised and evaluated for histopathology, mucus content, parietal and mucus cell counts. Blood was withdrawn from the orbital sinus of group B rats for biochemical evaluation (blood glucose, superoxide dismutase (SOD), lipid peroxidation (LP) and nitric oxide (NO)) and later sacrificed for gastric SOD, LP and NO evaluation. Blood glucose level was reduced (p<0.05) in all treatment groups compared to DU. Gastric SOD, parietal and mucus cell counts were increased (p<0.05) in the DMg250 and DI compared to DU. Serum LP and NO were reduced while gastric LP was increased in the DMg250 compared to DU. Gastric NO and mucous content were significantly reduced (p<0.05) in all diabetic groups compared to control. The gastric mucosa of the DU group had haemorrhage, inflammation and parasites embedded. The DMg250 and DI had normal submucus and muscle layers with reduced inflammation. Oral magnesium treatment in diabetes exerts hypoglycaemic effects, reduces serum nitric oxide and lipid peroxidation, increases gastric superoxide dismutase, mucous cell count and reduces the susceptibility of the gastric mucosa to ulceration.

Lipid Profile and Electrolyte Composition in Diabetic Rats Treated With Leaf Extract of Musa sapientum.

Diabetes mellitus affects lipid levels resulting in diabetic dyslipidemia as well as electrolyte loss from the body. Musa sapientum has been reported to possess antidiabetic properties. This study assessed the lipid profile and electrolyte composition in alloxan-induced diabetic rats treated with methanol leaf extract of M. sapientum (cMEMSL). Diabetes was induced with alloxan (120 mg/kg i.p.). Seventy-five male albino rats were divided into 5 groups of 15 rats each. Group 1 was control; groups 2-5 were made diabetic and treated with 0.2 ml 0.9% NaCl, cMEMSL (250 mg/kg and 500 mg/kg), and glibenclamide (5 mg/kg), respectively, for 14 days. Blood samples were obtained from the retro orbital sinus after light anesthesia from 5 animals in each group on days 2, 7, and 14 for lipids and electrolyte analysis. Lipid profile of diabetic treated (cMEMSL and glibenclamide) animals showed significant reduction (p < .05) in total cholesterol, triglyceride, and low density lipoprotein (LDL) levels. The high density lipoprotein (HDL) level in the treatment groups increased significantly (p < .05) compared with diabetic untreated. Sodium, potassium, and phosphate ions significantly increased in all diabetic treatment groups while chloride ion significantly decreased compared with diabetic untreated. There was no significant difference in calcium and bicarbonate ion concentration in all the groups. This study has showed additional properties of Musa sapientum to include its ability to restore electrolyte balance, reduce cholesterol, triglyceride, LDL, and increase the HDL levels in diabetic animals.

Anti-oxidative and reno-restorative effects of physalis angulata (whole plant extract) in alloxan-induced diabetic male Wistar rats.

Backgroiound: Hyperglycemia has been reported to increase protein glycation and generation of free radicals which predispose to diabetic renal dysfunction. Physalis ahgulata has been shown to have hypoglycacmic and anti-lipidemic properties but there is dearth of information regarding its effect on kidney functions in diabetes. This study investigated the anti-oxidative and reno-restorative effects of methanol extract of whole plant of Physalis angulata (MEPA) in alloxan-induced diabetic rats. METHODOLOGY: Twenty male Wistar rats (150-180g) were randomly divided into four groups: Group 1 (control) received 0.2 ml distilled water, groups 2-4 were made diabetic by single intra-peritoneal dose of alloxan monohydrate (100mg/kg) and treated with 0.2 ml distilled water, 500 mg/kg MEPA and 150 mg/kg metformin respectively. All treatments were given orally for 14 days. Blood samples were collected from each animal through retro-orbital puncture. The serum obtained were analysed for fructosamine, glycated hemoglobin (HbAlc), creatinine and blood urea nitrogen (BUN). Kidney samples were harvested into cold phosphate buffer, homogenized and centrifuged at- 7500rpm for 15 minutes. The supernatant obtained was analyzed for malondialdehyde and superoxide dismutase (SOD) activities. Values were compared using ANOVA at P<0.05. RESULTS: The MEPA-treated groups showed significant decrease (P<0.05) in blood glucose, kidney weights, fructosamine,. HbAlc, malondialdehyde, creatinine and BUN, while the body weights and SOD significantly increased (P<0.05) compared to diabetic untreated group. CONCLUSION: Treatment with methanol extract of Physalis angulata (whole plant) reduced hyperglycemia, malondialdehyde and glycation end- products, which could have contributed to the development of diabetic nephropathy if diabetes is left untreated.

Effects of quail egg on kidney functions in alloxan induced diabetic Wistar rats.

UNLABELLED: Abstract INTRODUCTION: Uncontrolled diabetes mellitus has been reported to lead to renal dysfunction. Quail egg consumption has been reported to exert curative effects in some disease conditions like diabetes mellitus, tuberculosis, and asthma. This study investigated the effects of quail egg consumption on some kidney functions in alloxan induced diabetic rats. METHODS: Forty male Wistar rats with an average weight of 170g were randomly divided into four groups. Groups A-Control, B-Diabetic untreated, C-Diabetic treated and D-Normal treated. Groups B and C were made diabetic with a single dose of alloxan monohydrate (100 mg/kg i.p). Raw quail egg was administered orally (5 ml/kg) to groups C and D for 14 days. Body weight and blood glucose were monitored during the study. Blood and kidney samples were obtained from animals in each group, and analyzed for total protein, creatinine, blood urea nitrogen (BUN), renal malondialdehyde (MDA) and superoxide dismutase (SOD). Data were analyzed using ANOVA at P < 0.05. RESULTS: Diabetic group treated with quail egg showed significant (P < 0.05) increase in SOD, decrease in body weight, blood glucose, total protein, creatinine, BUN and MDA levels when compared to diabetic untreated group. However, values of these parameters obtained from diabetic group treated with quail were comparable to control. CONCLUSION: Quail egg consumption significantly reduced hyperglycemia, serum total protein, creatinine, BUN, MDA and increased SOD activities in alloxan induced diabetic Wistar rats which suggests that it lowers blood glucose and ameliorates renal impairment in diabetes mellitus.

Anti-diabetic property of Methanol extract of Musa sapientum leaves and its fractions in alloxan-induced diabetic rats.

Diabetes mellitus is a metabolic disorder resulting from necrosis of ß-cell and insulin resistance at the cellular level. Musa sapientum has been shown to possess anti-diabetic properties, however, the mechanism of its action is unknown. The effect of Methanolic extract of Musa sapientum leaves (MEMSL) and its fractions were assessed for in vitro inhibitory activity of α-amylase enzyme, in vivo hypoglycemic properties and liver glycogen content in alloxan-induced diabetic rats. Dried plant powder of Musa sapientum was successively extracted using n-hexane, ethyl acetate, dichloromethane and methanol respectively. The filtrate obtained was evaporated using rotary evaporator and the extract was stored at 4°C until use. The methanolic extract obtained was further fractionated using column chromatography. In vitro alpha amylase inhibitory activity of the methanolic extract at different doses (2.5mg/ml, 5mg/ml, 10mg/ml, 25mg/ml and 50mg/ml) and column fractions (100ug/ml) were assessed and compared with that of acarbose (5mg/ml), a standard oral α-amylase inhibitor. Hypoglycemic activity and liver glycogen content was studied using alloxan -induced diabetic male rats treated with MEMSL (250mg/kg and 500mg/kg), column fractions F2 and F5 (100µg/kg) for 14 days respectively. Results obtained showed a dose -dependent increase in α-amylase inhibitory activity of the methanolic extract at 5, 10, 25 and 50mg/ml exhibiting 29%, 61%, and 72% and 80% inhibitory activities respectively. Column fractions 2 and 5 showed the highest α-amylase inhibitory activity of 79% and 74% respectively. The MEMSL at 250mg/kg and 500mg/kg exhibited 66% and 59% hypoglycemic activities respectively compared with diabetic controls. Fractions 2 and 5 showed 48% and 75% reduction in blood glucose level respectively. Liver glycogen in diabetic animals treated with MEMSL (250mg/kg and 500mg/kg), F2 and F5 were significantly increased (5.5±0.5, 5.9±0.7, 3.6±0.5, 8.0±0.4 mg/100gwt. liver) compared with Diabetic controls (1.2±0.3 mg/100gwt. liver) respectively suggesting an increase in glucose storage or reduction in glycogen breakdown. It seems possible that the anti-diabetic properties in the leaf extract of Musa sapientum and its fractions maybe due to the inhibition of α-amylase, increased storage of glucose as glycogen in the liver and/or reduced breakdown of liver glycogen stores.

Effect of magnesium on gastrointestinal transit time in normal and diabetic rats: possible mechanism of action.

BACKGROUND: Many gastrointestinal complications in diabetes are connected to neurohumoral dysfunction resulting in abnormalities of intestinal motility, secretion and absorption. Minerals have been reported as essential cofactors for basic cellular reactions but there is dearth of information on effect of Magnesium on gastrointestinal transit time (GITT) and the mechanism of action. METHODS: Sixty male albino Wistar rats (180 - 200g) were grouped into twelve of five animals each. Group 1 (control) received 0.2ml saline. Groups 2-6 were normal rats treated with magnesium sulphate (as magnesium) (500mg/kg), adrenaline (0.5mg/kg), magnesium (500mg/kg) and adrenaline (0.5mg/kg), prazosin (1mg/kg) and both magnesium (500mg/kg) and prazosin (1mg/kg) respectively. Groups 7 - 12 were diabetic rats treated as in groups 1- 6. Diabetes was induced intraperitoneally with alloxan (120mg/kg bwt). RESULTS: There was significant (p<0.05) reduction in GITT index in normal rats treated with magnesium, prazosin and combination of magnesium and prazosin compared with control. Treatment with adrenaline alone produced significant increase in GITT. However treatment with both magnesium and adrenaline produced significant reduction compared with control. This reduction in GITT was similar to that obtained in magnesium only and prazosin only treated groups. Diabetic groups showed significant reduction in GITT in all treated groups except the adrenaline only treated group which produced significant increase in GITT. CONCLUSION: The significant reduction in GITT produced by magnesium in both normal and diabetic animals was comparable to that produced by prazosin (an á-adrenoceptor antagonist) indicating that magnesium may be inhibiting gastrointestinal smooth muscle contraction through á-adrenoceptor antagonist pathway.

Effect of methanolic extract of Musa sapientum leaves on gastrointestinal transit time in normal and alloxan induced diabetic rats: possible mechanism of action.

Disorders of gastrointestinal motility have been associated with diabetes mellitus. Hyperglycaemia particularly has been reported to inhibit gastrointestinal transit time while glibenclamide, a sulphonylurea and insulin, both increased transit time. Musa sapientum has also been reported as an antidiabetic agent but there is dearth of information on the effect of this plant on gastrointestinal motility. This study was therefore carried out to investigate the effect of methanolic extract of Musa sapientum leaves (MEMSL) on gastrointestinal transit time (GITT) in male albino rats with and without hyperglycaemia and to elucidate possible mechanism by which this extract functions. Fifty five albino rats were divided into 11 groups of five animals each. All animals were fasted for 24hrs before the begining of the experiment. Group 1 served as control; while the remaining groups (2 - 11) were treated with 250mg/kg; 500mg/kg MEMSL; diabetic control; diabetic treated with 250mg/kg; 500mg/kg MEMSL; diabetic treated with glibenclamide (5mg/kg); normal rats treated with nifedipine (50mg/kg); normal rats treated with calcium chloride (CaCl2) only (10mg/kg); groups 10 and 11 were both pretreated with CaCl2 and subsequently treated with 250mg/kg and 500mg/kg MEMSL respectively. All plant extracts used for treatments were dissolved in normal saline and administered orally using orogastric tube. Charcoal meal was used as marker in the estimation of GITT. The study showed significant decrease in GITT in the normal rats treated with 250mg/kg and 500mg/kg of extract. However, in the diabetic rats treated with 500mg/kg MEMSL, there was significant increase in GITT and this is comparable with the gut response to glibenclamide (5mg/kg). Musa sapientum extract produced significant decrease in transit time in the calcium chloride pre-treated normal rats and this is comparable to the effect observed in Nifedipine treated group. The significant reduction in GITT produced by MEMSL in the normal rats reflects a strong possibility of MEMSL acting as calcium channel antagonist through the voltage gated calcium channel which may be due to the presence of alkaloids, saponins, cardenolides. There is the possibility of the extract acting as an inhibitor of potassium channel at higher concentration as observed in glibenclamide treated groups.

Effect of diet of varying protein concentrations on the activity of erythrocyte membrane Ca2+Mg2+ ATPase in dogs.

Alterations in protein diet have been reported to result in alterations in calcium homeostasis in the body. Ca2+Mg2+ATPase is an ubiquitous enzyme important in calcium homeostasis in the body. The effect of varying protein diet on the activities of Ca2+ pump across cell membranes is however yet to be fully elucidated. In this study, the activity of erythrocyte membrane calcium pump in response to varying protein concentration in diet was therefore studied in the dog. The study was carried out in 24 dogs, randomly divided into 4 groups. The groups were fed with diets containing 30%, 26%, 16% and 0% proteins (high, medium, low and zero) for six weeks respectively. Blood samples were collected from each animal to determine packed cell volumes, hematocrit, blood urea, electrolyte studies and erythrocyte ghost membrane studies. The effects of Ca2+ and ATP on the activity of Ca2+Mg2+ ATPase were determined in the isolated ghost membrane. The result of the study shows that there was a protein diet dependent increase in the activity of Ca2+Mg2+ ATPase in the presence and absence of ATP in all the groups with the highest activity recorded in the high protein diet group and the lowest activity observed in the zero protein group. There was also a protein diet dependent increase in the protein concentration of the membranes in all groups observed with the highest protein concentration recorded in the high protein diet group and the lowest activity observed in the zero protein group. There was a significant decrease in K+ concentration (P <0.05) and a significant increase in urea concentration of animals fed with high protein diet (P <0.05). There was also a significant increase (P <0.05) in HCO3- concentration in the animals fed with medium protein diet and no significant difference in the PCV and heamatocrit values in all groups. This study has shown that high protein diets increase the activity of the Ca2+Mg2+ ATPase in the presence and absence of ATP.

Anti-inflammatory activities of a kolaviron-inhibition of nitric oxide, prostaglandin E2 and tumor necrosis factor-alpha production in activated macrophage-like cell line.

Kolaviron (KV), a biflavonoid fraction from the seeds of Garcinia kola has been shown to posess antiinflammatory properties in animal models of inflammation. In this study, the effect of KV on carrageenan-induced paw edema was investigated in mice. Furthermore, the effects of KV on the production of the pro-inflammatory mediators- nitric oxide (NO) and tumor necrosis factor alpha (TNF-alpha) were examined in an activated macrophage-like cell lines, RAW 264.7 cells. Administration of KV prior to injection of carrageenan significantly reduced the paw inflammation in a dose-dependent manner. KV consistently inhibited in-vitro production of NO and secretion of TNF-alpha in a dose-dependent manner. In addition, KV reduced the production of PGE2 in LPS-stimulated macrophages. Viability of cells at all concentrations studied was unaffected as determined MTT [3-(4,5- dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide] cytotoxicity assay. These results suggest that KV has inhibitory effects on LPS-induced TNF-alpha, NO and PGE2 production.

Advances in the molecular understanding of GABA(B) receptors.

The molecular nature of the metabotropic GABA(B) receptor was for some time a mystery, however it was recently discovered that two related G-protein-coupled receptors have to heterodimerize to form the functional GABA(B) receptor at the cell surface. This review discusses the most recent findings in the rapidly expanding field of GABA(B) receptor research, and includes a summary of all splice variants of both receptor subunits identified to date. It also evaluates emerging evidence that certain splice variants might play a role in determining pharmacologically distinguishable receptors, and reviews receptor localization at the sub-cellular level and involvement in neuronal development.

Cellular and sub-cellular localisation of GABA(B1) and GABA(B2) receptor proteins in the rat cerebellum.

Following the recent discovery that GABA(B) receptors expressed in cell lines are only functional when both GABA(B1) and GABA(B2) are expressed, the present study reports on the development of polyclonal antisera specific for carboxyl-terminal portions of the two related GABA(B) receptor components respectively. Western blotting indicated the specificity of affinity-purified antibodies for native or recombinant expressed GABA(BR1) and GABA(BR2), with no cross-reactivity, both antisera detecting the heterodimer in rat cerebellar membranes. Immunohistochemistry revealed a distinct distribution of both receptor proteins in rat cerebellum. GABA(B1) immunoreactivity was primarily located in the granule cell layer and Purkinje cells, with discrete immuno-positive cell bodies being present in the molecular layer. GABA(B2) staining revealed intense immunoreactivity in the molecular layer, with weaker staining in the granule cell layer. Purkinje cell bodies were less intensely immuno-positive for GABA(B2). Co-localisation of both receptor proteins was observed using double immunofluorescence techniques, consistent with the notion that both proteins are required for the formation of functional GABA(B) receptors in vivo. Immunofluorescence also indicated that GABA(B) receptors did not co-localise with glial fibrillary acid protein, confirming a neuronal localisation for GABA(B) receptors. Electron microscopic analysis of the molecular layer revealed that the distribution of immunolabelling for both GABA(B1) and GABA(B2) was mainly located on the membrane of Purkinje cell dendrites and spines and in parallel fibre terminals.

GABA(B) receptor heterodimer-component localisation in human brain.

In recombinant cell lines, functional GABA(B) receptors are only formed by the heterodimerisation between two related G-protein coupled receptor proteins GABA(B)R1 (GBR1) and GABA(B)R2 (GBR2), whilst the individual GBR1 or GBR2 do not produce fully functional receptors. To determine whether the heterodimerisation occurs in vivo, novel polyclonal antibodies targeting the C termini of GBR1 and GBR2, were raised in different species, characterised, and used to determine the relative localisation of the reported heterodimer components in human brain tissue, using immunohistochemistry. The use of different species for the raising of the antisera allowed double immunofluorescent labelling of the receptors as an indication of GBR1/GBR2 receptor co-localisation in human brain. The presence of both proteins is reported in cerebellum, hippocampus, cortex, thalamus and basal ganglia. Regions of the brainstem including pons and medulla, also express GBR1 and GBR2 protein. The double immunofluorescence demonstrated that GBR1 and GBR2 are co-localised in the human cerebellar cortex. Together these results suggest the widespread distribution of GABA(B) receptors in human brain, and that GABA(B) receptors GBR1 and GBR2 can exist in the same cell, and therefore may function as a heterodimer in the human brain.

African histoplasmosis (Duboisii) of the skull with neurological complication--a case report and review of literature.

A case of African histoplasmosis of the skull associated with neurological deficit has been reported. There was complete recovery of neurological features after excision of the lesion followed by a course of co-trimoxazole. A review of the available literature indicates the rarity of this particular mode of presentation. The reversibility of the neurological complications makes it important that clinicians increase their awareness of this treatable condition.