39 | Okereke Stanley Chukwuma1,Nwaogwugwu, Joel Caleb Chukwudoruo ,Chieme Sunday. Year of Publication- 2017 | In-Vitro Antioxidant Activity of Methanolic Leaf Extract of Tridax ProcumbensandTithoniaDiversifolia | | Journal of Medical Science and Clinical Research.(JMSCR) | Volume05/Issue 05/ Page: 21783-21789.
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40 | SC Okereke, UO Arunsi, AC Ngwogu, JO Onyike, C.S Chukwudoruo.Year of Publication- 2018. | Identification of the Active Ingredients in Methanolic Leaf Extract of Aspiliaafricana(Pers.)C.D. Adams and their Effects on Ibuprofen Induced Ulcer Model in Wistar Rats | Peptic ulcer is one of the most dreadful diseases in the world today. However, the present study was investigated to identifythe active ingredients in the methanolic leaf extract of Aspiliaafricana and their possible effects on Ibuprofen induced ulcer model inWistar rats using standard analytical methods. The GC-MS spectral analysis revealed thirty three (33) active ingredients with 9,12,15-Octadecatrienoic acid, methyl-ester(z,z,z)-, stigmasterol, hexadecanoic acid, methylester(z)-, Methyl linolelaidate, n-Hexadecanoic acid,squalene, phytol and vitamin E possessing important pharmacological and cytoprotective functions. The oral acute (LD50) toxicity study divulged that the extract did not cause mortality to any of the experimental animals even at the highest dose of 5000mg/kg. Other studies disclosed that the induction of ulcerogenesis by ibuprofen led to alterations in the mucosal membrane and this was evidenced by severe distortions in gastric contents, serum concentrations of blood glucose, antioxidant enzymes, lipid peroxidation marker, hepatocellular enzymes, renal function and haematologicalindexes. Pretreatment with the natural antidote (A. africana leaf extract) remediated this imbalance significantly more than a standard antiulcer drug (omeprazole). From these observations, we conclude that the methanolic leaf extract of A. africana offers more promising gastroprotection than omeprazole and due to low toxicity; it should be incorporated in traditional medicine for the management and treatment of peptic ulcer disease. Keywords: Aspiliaafricana, active ingredients, NSAIDs, antiulcerogenicity, GC-MS. | International Journal of Science and Research. | Volume 7,Issue 1,Page: 310-319.
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41 | Linus A. Nwaogu*., Chidi U. Igwe., Kingsley U. Obasi and Ujunwa L. Okeke (2017) | Chemical Composition of Siegesbeckia orientalis- A Valuable, But Less Known Ethnomedicinal Plant. | The chemical (phytochemical, vitamin, elemental and proximate) compositions of the leaves of Sigesbeckia orientalis was determined using standard methods. The results of the phytochemical composition in mg/100g indicated that saponins appeared to be the highest followed by alkaloids, flavonoids, tannins, oxalates, cynogenic glycosides and phytate. The leaf contains appreciable concentrations in mg/100g of vitamins A and C. The leaves contain vital concentrations in mg/100g of minerals which include calcium, magnesium, phosphorus, iron and zinc. It also has trace amount of potassium, sodium, manganese, copper and cobalt. The moisture content of the leaf was 75.50±1.00 %, ash 4.77 ± 0.03%, proteins 10.73 ± 0.03%, fiber 6.38 ± 0.11%, fats 2.27±0.07% and carbohydrate 0.35±0.02% respectively. The study revealed that Sigesbeckia orientalis leaf is a good source of vital vitamins and minerals. It also contains important phytochemicals which have been reported to have various biochemical and physiological effects which confer its medicinal benefits to users. Key words: Ethnomedicinal plant, phytochemical, proximate, mineral, Sigesbeckia orientalis | Biokemistri. Journal of Nigerian Society for Experimental Biology | 29 (2): 54-60
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42 | Emejulu A.A,AlisiA.P, Onwuliri, V.A. and AwurumC.S. 2017 | Effect of Ethanol Leaf Extract of Jatropha Curcas on Some Liver and Kidney Function Biomarkers of Normal Rabbits | This study investigated effects of ethanol leaf extract of Jatropha curcas on liver and kidney function biomarkers of male New Zealand rabbits. A total of 20 rabbits were divided into four groups of 5 animals each. Graded concentrations of ethanol extract of the leaf administered thus: 50 mg, 150 mg, 250 mg and 0 mg/kg bodyweight of animals in groups I, II, III and IV (control) respectively for 10 days by oral intubation. Serum liver enzymes and renal function were assessed using standard procedures. Alkaline phosphatase (ALP) activity of all treated rabbits significantly increased (p<0.05) in a dose-dependent manner. There was also a significant elevation in activity of aspartate aminotransferase (AST) of rabbits treated with 250 mg/kgbwt (31.0 ± 5.48 U/l) compared to control (10.00 ± 2.44 U/l). However, no significant difference was observed in alanine aminotransferase (ALT) activity. Conjugated bilirubin concentration increased significantly (p<0.05) in a non-dose dependent manner. Serum urea concentration of the rabbits treated with 250 mg/kg bodyweight of leaf extract (11.90 ± 0.24 mmol/l) was significantly (p<0.05) higher than that of the control (8.01 ± 0.89 mmol/l), while serum creatinine concentration increased significantly (p<0.05) in groups treated with 150 and 250 mg/kg body weight. Serum potassium concentration of rabbits showed a significant increase (p<0.05) at 150 mg/kg and 250 mg/kg bodyweight, while serum sodium significantly increased (p<0.05) in a non-dose dependent manner. These results suggest that ethanol extract of J.curcas has some hepatobiliary toxic effect, as well as renal dysfunction Keywords: Hepatic function, Renal function, Jatropha curcas, Rabbits, Phytochemicals. | FUTOJNLS 2017 Volume-3, Issue-2, pp- 213 - 224 | Futo Journal Series
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43 | AA Emejulu, KC Nwufo, AC Ene, UK Obasi.2017 | GC-FID Phytochemical Analysis and Intraperitoneal Lethal Dose (LD50) Determination of Ethanol Root Extract of Mucunapruriens | The study was designed to evaluate the qualitative and quantitative phytochemical profile and determine the lethal dose (LD50) of ethanol root extract ofMucunapruriens. This plant is extensively used in the South Eastern Nigeria for the management of certain ailments such as Parkinson's disease, diabetes mellitus and anaemia. The quality of the phytochemical contents was determined using standard analytical methods, while a Flame ionization detector equipped Gas chromatography (GC-FID) was used for further identification and quantification of the phytochemicals. The lethal dose (LD50) was also established. Preliminary phytochemical screening revealed the presence of Alkaloids, flavonoids, tannins, saponins and phenols. Further analysis with GC-FID revealed the presence of saponins (17.6643μg/ml), sapogenin (11.8864μg/ml), catechin (4.2495μg/ml), kaempferol (3.6267μg/ml), rutin (2.5745μg/ml), anthocyanin (0.6381μg/ml), ribalinidine (73.9745μg/ml), lunamarine (2.3099μg/ml), phenols (2.7962μg/ml), tannins (5.1494μg/ml), oxalate (4.3036μg/ml), phytate (0.1891μg/ml) and spartein (0.0001μg/ml). The intraperitoneal lethal dose (LD50) value for the root extract was found to be 2154mg/kg body weight. Mucunapruriens ethanol root extract contains therapeutically useful phytochemicals with considerably safe lethal dose value. Keywords: GC-FID Phytochemical Analysis, LD50, Keywords: GC-FID Phytochemical Analysis, LD50, Mucunapruriens, Intraperitoneal, root extract Mucunapruriens, Intraperitoneal, root extract. | International Journal of Research in Pharmacy and Biosciences Volume 4, Issue 7, 2017, PP 23-28 | Sryahwa Publications
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44 | Igwe C, Emejulu A, Nwankwo C, Onwuliri V. 2017 | Alterations in lipid profile and atherogenic predictor indices of type 2 diabetes mellitus patients in Awka, Anambra State, Nigeria. | Changes in lipid profile and atherogenic predictor indices of Type 2 diabetes mellitus (T2DM) patients attending Diabetic Clinic of ChukwuemekaOdumegwuOjukwu University Teaching Hospital Awka, Anambra State, Nigeria, between January 2016 and January 2017 were studied using standard methods. Fasting blood glucose (FBG), total cholesterol (TC), triglyceride (TG) and high density lipoprotein cholesterol (HDL-c) concentrations were determined spectrophotometrically, while the values of low density lipoprotein cholesterol (LDL-c), very low density lipoprotein cholesterol (VLDL-c) and the atherogenic lipid ratios were calculated. The subjects included willingly consented 60 T2DM patients and 40 apparently healthy, age-matched control subjects. The FBG concentration, lipid profile and atherogenic parameters were significantly (p<0.05) higher in the diabetics than the control subjects. Similar trend of results were observed within the male and female subgroups of the subjects. Majority of the diabetic subjects had abnormally high prevalence of dyslipidaemia. Over 60% of the diabetic subjects had high levels of atherogenic predictor indices in the range of patients with abnormal Castelli’s Risk Index-II >Atherogenic Index of Plasma > non HDL-c >Atherogenic Coefficient >Castelli’s Risk Index-I. FBG concentration correlated significantly (r=0.28; p=0.027) with non HDL-c of the diabetic subjects. The occurrence of dyslipidaemia and abnormality in atherogenic risk predictor indices among Nigerian T2DM patients is high. The significantly high values of lipid profile and atherogenic indices obtained might indicate potential and increased risk of coronary heart disease (CHD) among these patients. Thus, the need for increased awareness for regular lipid profile testing, monitoring and use of atherogenic predictor indices in the effective management of T2DM patients is very crucial. | Biokemistri, Vol 29, No 2 | Biokemistri |
45 | KC Nwufo, AC Ene, AA Emejulu, UK Obasi, CU Ene 2017 | Antidiabetic Properties of Ethanolic Root Extract of MucunaPruriens on Alloxan Induced Diabetic Rats | The study was aimed at evaluating the antidiabetic properties of ethanol root extract ofMucunapruriens. Ethanol root extract of Mucunapruriens was studied for its in vivoantidiabetic potentials using alloxan induced diabetic rats. Glucose tolerance test was performed using both diabetic and normal rats. Different groups of diabetic rats were subjected to daily intraperitoneal administration of Mucunapruriens at 100, 150 and 200mg/kg body weight for 14 days. Blood samples were collected at the end of the 14days treatment for biochemical and hematological examinations. Glucose tolerance curve showed a significant decrease in blood glucose concentrations of rats treated with a single dose of Mucunapruriens(300mg/kg body weight) compared to the untreated controls. Treatment of alloxan induced diabetic rats with 100 and 150 mg/kg body weight ofMucunapruriens for 14 days significantly (p<0.05) reduced fasting blood glucose concentration to normal from day 5. Following induction of diabetes mellitus, the serum activities of Aspartate amino transferase (AST), Alanine amino transferase (ALT) and Alkaline Phosphatase (ALP) and the serum concentrations of urea, creatinine, total cholesterol, low density lipoprotein cholesterol (LDL-C) and triglycerides were significantly (p<0.05) elevated with concomitant reduction in the serum concentrations of total protein, albumin, Na ion, K ion, bicarbonate ion and high density lipoprotein cholesterol (HDL-C), compared to the normal control. Treatments with different doses of Mucunapruriensandglibenclamide restored these parameters to basal level. There were serious decrease in all the hematological parameters of the diabetic untreated rats, however, these changes were restored to near normal with treatments with Mucunapruriens. This study therefore suggests that the ethanol root extract of Mucunapruriensstronglypossesantidiabetic properties and could be a potential source of treatment for diabetes mellitus. Keywords: Diabetes mellitus, Mucunapruriens, glucose tolerance, hematology, antidiabetic | International Journal of Research in Pharmacy and Biosciences Volume 4, Issue 5, 2017, PP 17-29 ISSN 2394-5885 (Print) & ISSN 2394-5893 (Online) | Sryahwa Publications
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46 | Ezeji-Chigbu NGN, EneAC, Emejulu AA.2018 | Sub-Acute Toxicity Evaluation of Aqueous Stem Bark Extract of SarcocephalusLatifolius | Aqueous extract of the stembark of Sarcocephaluslatifolius was administered to wistar albino rats to assess sub-acute toxicity. Twenty-four wistar albino rats were divided into four groups of six animals per group. Three groups were administered increasing doses of the extract 125, 250 and 500 mg/body weight whereas the fourth group was used as normal control. The duration of the experiment was two weeks. This study revealed that the aqueous extract of stem bark extract of Sarcocephaluslatifolius affected the biochemical parameters at the test doses of 125, 250 and 500 mg/body weight. The biochemical parameters studied showed some consistent changes. There was an elevation in ALT, ALP, AST, Total Bilirubin, Direct Bilirubin, Urea, Creatinine, Bicarbonate, Chloride, Potassium, Cholesterol and Triglyceride levels in the treated groups when compared to the control groups. There were statistically significant difference (p<0.05) observed when these biochemical parameters in the treated groups were compared with the normal control. This increase in biochemical parameters was higher as the doses of the aqueous extract of the stem bark of Sarcocephaluslatifolius increased. On the other hand, there was a decrease in the concentrations of Protein, Albumin, Sodium and HDL-Cholesterol in the treated groups when compared to the Control group. This decrease was observed to be statistically significant (p<0.05) when these biochemical parameters in the treated groups were compared with the normal control. There was no statistically significant difference (p>0.05) observed in the hematological parameters of the test animals compared to the control. The histopathology results of the sub-acute toxicity of the aqueous extract of the stem bark of Sarcocephaluslatifolius concurs with the results of the biochemical parameters. From the result, there were no severe pathological changes in the cell architecture of the liver, kidney and heart of the extract treated animals. These results indicate that the extract may be toxic at higher doses and short term exposure. KeywordsSarcocephaluslatifolius; Aqueous Extract; Albino Rats; Sub-Acute Toxicity | SciFed Journal of Analytical Biochemistry 1:1. | SciFed Publishers
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47 | Ene AC, EzejiChigbu NGN, EmejuluAA,Ene CU, Okwu GN, Ujowundu CO 2018 | Antiplasmodial and Antioxidant Evaluation of the Methanol and Aqueous Extracts of SarcocephalusLatifolius | The methanol and aqueous extracts of different parts of Sarcocephaluslatifolius plant were studied for their in vivo anti-plasmodial activity against Plasmodium bergheiNk 65in Swiss albino mice. Phytochemicals and Antioxidant studies were also conducted for all the plant extracts used. The aqueous extract of the stem bark of Sarcocephaluslatifolius showed the highest antiplasmodialactivity.The extract significantly suppressed the parasite. There was a significant difference (p<0.05) observed when the level of parasitemia of the animals treated with the aqueous extract of the stem bark of Sarcocephaluslatifolius was compared with the untreated, chloroquine standard control and other treatment groups. Detailed phytochemical screening indicated that the crude extracts contained secondary metabolites including phenols, tannins, alkaloids, saponins, steroids, phytate, oxalate, cyanide and anthraquinone. The antioxidant studies showed that the methanol extract of the stem bark has significantly the highest percentage activity of 2,2-diphenyl-1-picrylhydrazyl (DPPH). The aqueous extract of Sarcocephaluslatifolius yielded more of the secondary metabolites compared to the methanol extracts. The in vitroantiplasmodial activity exhibited by these extracts might be attributed to these secondary metabolites. Since Sarcocephaluslatifolius showed very good antiplasmodial activity, it is concluded that this plant has potentials in fighting malaria. Keywords AntiplasmodialActivity; Sacocephaluslatifolius; Plasmodium berghei; Albino mice | SciFed Journal of Analytical Biochemistry 1:1. | SciFed Publishers
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48 | Chikezie C.M, Ojiako O.A, Emejulu A.A. and Chikezie P. C. 2018 | Serum Lactate Dehydrogenase Activity and Visceral Organs and Body Weights of Diabetic Rats Administered Single and Combinatorial Herbal Formulations. | Background and Aim: The present study evaluated serum lactate dehydrogenase (LDH) activity. Visceral organs and body weights were used as measures of the capacities of single and combinatorial herbal formulations of Acanthus montanus, Asystasiagangetica, Gongronemalatifolium and Solanummelongenas to ameliorate systemic toxicity, visceral organs inflammation or necrosis and body tissues wasting in alloxan-induced diabetes mellitus (DM) rats. Materials and Methods: Alloxan-induced DM rats with fasting plasma glucose concentration (FPGC) > 5.71 mmol/L for 5 consecutive days were selected for the study. A total of 102 male Wistar rats were divided into seventeen (17) groups of six (6) rats each. Serum LDH activity and body weights and weights of visceral organs and were measured using standard methods. Results: Serum LDH activities of herbal treated rat groups varied within a relatively narrow range of 549.9 ± 12.10 – 500.6 ± 12.02 IU/L and were significantly lower (p < 0.05) than the untreated DM rat group. The body weights of the experimental rat groups after herbal treatment were significantly higher (p < 0.05) than their corresponding weights before herbal treatment. The ratios of liver weights to body weights were within the range of 0.0293 ± 1.4 x 10-3 – 0.0597 ± 2.3 x 10-3. The ratio of kidney weight to body weight of untreated DM rat group was 1.64 fold higher than that of normal rat group (p > 0.05). Conclusion: Overall, 200 mg/kg body weight double herbal formulations of A. gangetica + A. montanus and A. gangetica + G. latifolium offered the greatest therapeutic benefits to alloxan-induced DM rats, with respect to all diagnostic parameters considered in the present study. Keywords:Body weights, Diabetes mellitus, Herbal formulations, Lactate | Pharmacogn. Commn. 8(1): 37-42 | Pharmacognosy Communications
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49 | Chikezie C.M, Ojiako O.A, Emejulu A.A. and Chikezie P. C. | Atherogenicity of diabetic rats administered single and combinatorial herbal Extracts | Objective: The present study sought to investigate atherogenicity of alloxan-induced diabetic rats administeredsingle and combinatorial herbal formulations of Acanthus montanus, Asystasiagangetica, Gongronemalatifoliumand Solanummelongena. Methods: A single intra-peritoneal (i.p.) injection of 90 mg/kg b.w. of alloxan monohydrate was given to the ratsto induce diabetes mellitus (DM). Serum lipid profiles were measured using standard spectrophotometricmethods, whereas atherogenicity, serum lipid ratios and atherogenic coefficient/indices were calculated usingstandard formulae. Results: Serum total cholesterol (TC) concentrations of experimental rat groups varied between1.59 ± 0.10 mmol/L and 2.72 ± 0.16 mmol/L (p < 0.05). Serum high-density lipoprotein cholesterol (HDLC)concentration of untreated DM rat (DM-r) group was significantly lower (p < 0.05) than those of treated DMrgroups. Atherogenic risk indices (ARIs) of treated DM-r groups were within the range of 0.74 ± 0.03 and2.64 ± 0.21, whereas ARI of untreated DM-r was 4.04 ± 0.25. The linear regression analysis of atherognicindex of plasma (AIP) versus serum low-density lipoprotein cholesterol (LDL-C) concentrations of the experimentalrat groups gave a relatively close fitted regression line (R2=0.8275). Atherogenic protection of herbalextract treated DM-r groups was within the range of 33.4–81.7%. Conclusion: The present study showed that double herbal formulations (DHFs): A. gangetica+G. latifolium andA. gangetica+A. montanus offered comparatively high protection to DM-r against atherogenic outcomes, whichparalleled the capacities of these DHFs to reverse dyslipidemia. | Bulletin of Faculty of Pharmacy, Cairo University 56 | Faculty of Pharmacy, University of Cairo.
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50 | Ibeawuchi-Onuoha, Chinyere (2017) | Cell cycle distribution response of human lymph node prostate carcinoma (LNCaP) cell line to selected diet-derived agents | Decline in prostate cancer incidence and mortality rates can be achieved with the adoption of healthier lifestyles and chemoprevention. The use of some notable diet-derived agents has been advocated based on cultural inclinations and basic scientific outcomes. However significant advances in chemoprevention can be made chiefly through the comprehensive knowledge of these naturally-sourced compounds. Alterations to the cell cycle mechanism of LNCaP cell lines were investigated for responses to selected doses of curcumin, 3,3’- diindolylmethane (DIM) and epigallocatechin-gallate (EGCG). The flow cytometry technique in combination with propidium iodide (PI) staining was used for the study. Results obtained reveal different levels and occurrences of cell cycle arrest. The understudied diet-derived agents possess the capabilities of inhibiting prostate cancer growth and survival by inducing cell cycle arrest although at varying degrees, which needs further investigations. | Biokemistri | Vol. 29 (no. 4, 2017) pp 146–150
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51 | Ibeawuchi-Onuoha, Chinyere (2017) | Metabolomic analysis of epigallocatechin-gallate (EGCG)–treated human lymph node prostate carcinoma (LNCaP) cell line | EGCG (Epigallocatechin-gallate) is frequently studied and the observed effects show potentials in prostate chemoprevention. The mechanism of action EGCG has on prostate cancer cells is still not fully understood.However, through metabolic profiling, the effect of EGCG on prostate cancer cells could be elucidated and potential metabolic targets identified. Here, the metabolic profile of EGCG treated LNCaP cells (n = 6) were compared to untreated controls (n = 6). Comparatively, there were significant changes in the metabolic levels of lactate, citrate, aspartate, succinate, asparagine, glycine and adenine nucleotides. The findings suggest that the polyphenol EGCG is effective in deregulating the metabolism of prostate cancer cells by targeting the Krebs cycle. | Biokemistri | Vol. 29 (no. 1, 2017) pp 12–18
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52 | Iheme CI, Omeh YN and Nwuke CP | Variation effects of mobile phone frequencies and exposure durations on selected oxidative stress biomarkers of male wistar albino rats | The effects of different frequencies and exposure duration on selected oxidative stress biomarkers of male albino wistar rats were studied. Sixty (60) rats were used for the study. The rats were divided into two major groups of thirty (30) rats each, and each major group were further divided into sub-groups, I-VI, of five (5) rats each according to their weights. The rats in major groups, A and B, were exposed for ninety (90) and one hundred and eighty (180) days respectively. The rats in sub-group I, II, and III were exposed to 900 MHz field from mobile phones for 5 h/day, 10h/day, 15 h/day respectively, while the rats in subgroup IV were exposed to 2.4 GHz field for 24 h/day. Rats in subgroups V and VI were the normal and negative controls respectively. At the end of 90 and 180 days, the rats were anaesthetized; blood samples were collected for assessment of oxidative stress biomarkers. The activities of SOD, CAT and the concentration of MDA from subgroup III were 12.69 ± 0.60, 14.74 ± 0.07, and 2.03 ± 0.10 respectively. The activities of SOD, CAT and the concentration of MDA at these varying exposure time for 90 days duration were significantly (p<0.05) higher than those of positive and negative controls which were 8.64 ± 0.35, 9.60 ± 0.47, 1.03 ± 0.02 and 9.04 ± 0.02, 10.43 ± 0.51, 1.20 ± 0.20 respectively. At the exposure duration of 180 days, the activities of the SOD, CAT and the concentration of MDA were assessed. The activities of SOD, CAT and the concentration of MDA from subgroup III were 13.47 ± 0.51, 14.84 ± 0.07, and 2.11 ± 0.10 respectively. The activities of SOD, CAT and the concentration of MDA at these varying exposure time for 180 days were significantly (p<0.05) higher than those of positive and negative controls which were 8.91 ± 0.10, 9.94 ± 0.13, 1.18 ± 0.19; and 9.09 ± 0.03, 10.45 ± 0.49, 1.38 ± 0.20 respectively. At the different frequency of 2.4 GHz, the activities of the enzyme and the concentration of MDA were also assayed. At the exposure of 90 days duration, the activities of SOD, CAT and the concentration of MDA were 14.49 ± 0.29, 15.77 ± 0.10, and 2.67 ± 0.04 respectively. At the exposure of 180 days duration, the activities of SOD, CAT and MDA were 14.96 ± 0.10, 15.86 ± 0.11, and 3.00 ± 0.08 respectively. In general, the activities of SOD, CAT and MDA at the frequency of 2.4 GHz for the exposure duration of 90 and 180 days were significantly (p<0.05) higher than those positive and negative controls. From our findings, it can be concluded that different frequencies (900 MHz and 2.4 GHz) and exposure durations (90 and 180 days), can induce oxidative stress in living system. | Biochemistry & Analytical Biochemistry | 7/3
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53 | Callistus I Iheme*, Doris I Ukairo, Chiedozie O Ibegbulem, Olivia O Okorom and Kelechi Chibundu | Analysis of Enzymes Activities on Domestic Waste Dump Sites | Introduction: Effects of physico-chemical parameters on microbial dehydrogenases from domestic waste dumpsites were studied. Methodology: The microorganisms (S. aureus, E. coli, P. aeruginosa, C. albicans, T. Mentagrophytes and F. oxysoprium) were isolated from three domestic waste dumps in Owerri metropolis and subculture. Microbial dehydrogenases were extracted with acetone, partially purified by ammonium sulphate precipitation, dialysis, DEAEcellulose column chromatography and Sephadex G200 gel filtration chromatography. Dehydrogenase activity at each stage of purification was assayed with 2,3,5-triphenyltetrazolium chloride (TTC) and the absorbance of the formazan formed was measured and used to ascertain the total dehydrogenase activity. The impacts of effectors (Ca2+, Mg2+, Zn2+, Fe2+ and EDTA, ethanol and butanol) on the microbial dehydrogenase were determined by incubating the partially purified enzyme with the effectors for 30 min at 4°C. Also, the effects of temperature and pH were assessed by varying the temperature and pH ranges from 10°C to 60°C and 2.0 to 8.0, respectively. Results: The specific activities of the enzyme from the microorganisms were 7.10, 7.73, 6.47, 6.26, 9.66 and 10.58 mg Formazan/mg cell dry wt/h, respectively. Calcium ion, Mg2+, ethanol and butanol significantly increased (p<0.05) dehydrogenase activities in all the microorganisms studied while Zn2+, Fe2+ and EDTA decreased the activities. Conclusion: These activators can be harnessed within the recorded optimum temperature and pH ranges to enhance microbial growth; which is essential for the degradation of domestic waste thereby promoting cleaner environment. | Journal of Bioremediation & Biodegradation | 4-Aug
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54 | Chidi Uzoma Igwe*, Linus A. Nwaogu, Emeka E. Ezeokeke, Callistus I. Iheme, and Love Nma Alison | Effect of Ethanol Leaf Extract of Moringa oleifera on Oxidative Stress and Atherogenic Indices of Otapiapia-Exposed Albino Rats | Objective: Otapiapia is a locally produced, easily accessible, non-regulated household pesticide, which could easily contaminate human food. The ameliorative effect of ethanol leaf extract of Moringa oliefera on otapiapiainduced changes in oxidative stress, lipid profile and atherogenic indices of albino rats was investigated using standard methods. Methodology: Twenty-four apparently healthy Wister male albino rats (150-180 g) were divided into 4 groups (IIV) of 6 animals each. Group I served as the control, while group II animals were given otapiapia contaminated feed and distilled water. Group III were fed uncontaminated diet and M. oleifera extract drink. Group IV animals were also given the contaminated diet and M. oliefera extract. The feed and drinks were provided ad libitum for a period of 14 days. Result: The results show that intake of otapiapia significantly (p<0.05) reduced serum superoxide dismutase and catalase activities and HDL-c concentration but increased significantly (p<0.05) the concentrations of malondialdehyde, total cholesterol, triacylglycerol, LDL-c and VLDL-c. The derangements were significantly more pronounced in the atherogenic predictor indices than with lipid profile values. The observed effects of otapiapia were significantly (p<0.05) countered by administration of M. oliefera leaf extract. Conclusion: The results indicate that M. oleifera is a potent anti-poison with ameliorative effect against otapiapia-induced changes in biomarkers of oxidative and atherogenic damages in animals. | Journal of Clinical Toxicology | 3-Jul
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55 | C. O. Ujowundu, G. N. Okw, J. J. Achilike, L. A. Nwaogu, A. C. Ene and C. I. Iheme | Lead induced Oxidative Stress and Chemoprotective Role of Dietary Supplements on Wistar Albino Rats | The heavy metal lead (Pb) is a common environmental pollutant with widespread distribution, and oxidative stress has been implicated in the pathogenesis of its toxicity. The ameliorative effect of nutritional contents of palm oil and cod liver oil (dietary supplements) following exposure to sublethal concentration of Pb on adult Wistar albino rats was studied. Toxicity was induced by administering intraperitoneally, 30 mg/kg body weight of lead acetate at alternate days for 21 days. Groups treated with supplements received daily oral dose of 2.5 ml palm oil or cod liver oil or 1.25 ml palm oil and 1.25 ml cod liver oil (synergy). Increased activities of alanine aminotransferase, aspartate aminotransferase, alkaline phosphatase, decreased activities of superoxide dismutase, catalase, glutathione peroxidase and increased concentrations of total bilirubin and lipid peroxidation product were observed in Pb exposed rats without nutritional supplement treatment. However, these negative oxidative states were ameliorated in rats by the concomitant administration of nutritional supplements, singly and in combination. Furthermore, non-significant changes were observed in the haematological parameters determined. These observations indicate potential therapeutic benefits in the use of palm oil and cod liver oil in the management of lead-induced toxicity. | Annual Research & Review in Biology | 13/6
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56 | Nwufo, K.C., *Ene, A.C., Emejulu, A.A. Obasi, U.K. and Ene C.U. (2017) | Antidiabetic Properties of Ethanolic Root Extract of Mucuna Pruriens on Alloxan Induced Diabetic Rats | The study was aimed at evaluating the antidiabetic properties of ethanol root extract ofMucunapruriens. Ethanol root extract of Mucunapruriens was studied for its in vivoantidiabetic potentials using alloxan induced diabetic rats. Glucose tolerance test was performed using both diabetic and normal rats. Different groups of diabetic rats were subjected to daily intraperitoneal administration of Mucunapruriens at 100, 150 and 200mg/kg body weight for 14 days. Blood samples were collected at the end of the 14days treatment for biochemical and hematological examinations. Glucose tolerance curve showed a significant decrease in blood glucose concentrations of rats treated with a single dose of Mucunapruriens(300mg/kg body weight) compared to the untreated controls. Treatment of alloxan induced diabetic rats with 100 and 150 mg/kg body weight ofMucunapruriens for 14 days significantly (p<0.05) reduced fasting blood glucose concentration to normal from day 5. Following induction of diabetes mellitus, the serum activities of Aspartate amino transferase (AST), Alanine amino transferase (ALT) and Alkaline Phosphatase (ALP) and the serum concentrations of urea, creatinine, total cholesterol, low density lipoprotein cholesterol (LDL-C) and triglycerides were significantly (p<0.05) elevated with concomitant reduction in the serum concentrations of total protein, albumin, Na ion, K ion, bicarbonate ion and high density lipoprotein cholesterol (HDL-C), compared to the normal control. Treatments with different doses of Mucunapruriensand glibenclamide restored these parameters to basal level. There were serious decrease in all the hematological parameters of the diabetic untreated rats, however, these changes were restored to near normal with treatments with Mucunapruriens. This study therefore suggests that the ethanol root extract of Mucunapruriensstrongly possesantidiabetic properties and could be a potential source of treatment for diabetes mellitus. Keywords: Diabetes mellitus, Mucunapruriens, glucose tolerance, hematology, antidiabetic | International Journal of Research in Pharmacy and Biosciences | Volume 4, Issue 5, PP 17-29
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57 | Ibegbulem CO, Ene AC, Nwanpka P, Chikezie PCand Igwe CU. (2017) | Effect of Wood Charcoal Powder on Rate of Microbial Production of Lactic Acid in Dehulled and Undehulled Vigna unguiculata Pastes | Effect of Wood Charcoal Powder (WCP) on rate of microbial production of Lactic Acid (LA) in ground dehulled and an undehulled Vigna unguiculata (cowpea) paste was studied. The pastes were analysed for proximate compositions, energy values, tannin contents and microbial loads. Later, 4.0 g pastes were treated with incremental concentrations (0.03-0.05 g) of WCP diluted with 50 mL distilled water and fermented for 2 h at ambient temperature (29.5 ± 2.0ºC). Results showed that decortication of cowpea significantly reduced (p<0.05) the crude protein, crude fibre, crude fat and tannin contents as well as energy value and microbial loads. Fermentation produced ethanol and LA. The WCP inhibited LA production in dehulled cowpea paste, whereas LA fermentation continued unhindered in Undehulled Cowpea Paste (UCP). A WCP-tannic acid-cellulose interaction study suggested that WCP interacted more with the fibre and tannin components of UCP thereby reducing the amount that would have bound the fermentative microorganisms. In conclusion, wood charcoal inhibited microbial production of lactic acid in ground dehulled cowpea paste. | Journal of Nutrition & Food Sciences | 7:02:00 AMDOI: 10.4172/2155-9600.1000587
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58 | C. O. Ujowundu1, G. N. Okwu, J. J. Achilike, L. A. Nwaogu1, A. C. Eneand C. I. Iheme (2017) | Lead-induced Oxidative Stress and Chemoprotective Role of Dietary Supplements on Wistar Albino Rats | The heavy metal lead (Pb) is a common environmental pollutant with widespread distribution, andoxidative stress has been implicated in the pathogenesis of its toxicity. The ameliorative effect of nutritional contents of palm oil and cod liver oil (dietary supplements) following exposure to sublethal concentration of Pb on adult Wistar albino rats was studied. Toxicity was induced by administering concentration of Pb on adult Wistar albino rats was studied. Toxicity was induced by administering with supplements received daily oral dose of 2.5 ml palm oil or cod liver oil or 1.25 ml palm oil and 1.25 ml cod liver oil (synergy). Increased activities of alanine aminotransferase, aspartate aminotransferase, alkaline phosphatase, decreased activities of superoxide dismutase, catalase, glutathione peroxidase and increased concentrations of total bilirubin and lipid peroxidation product were observed in Pb exposed rats without nutritional supplement treatment. However, these negative oxidative states were ameliorated in rats by the concomitant administration of nutritional supplements, singly and in combination. Furthermore, non-significant changes were observed in the haematological parameters determined. These observations indicate potential therapeutic benefits in the use of palm oil and cod liver oil in the management of lead-induced toxicity. | Annual Research & Review in Biology | 13(6): 1-14
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59 | Ezeokeke EE, Ene ACand Igwe CU. (2017) | Sub-Acute Toxicity Studies of Alchornea cordifolia Leaf Extract in Swiss Albino Rats | The ethanolic leaf extract of Alchornea cordifolia (Schum. and Thonn.) Müll. Arg (Euphorbiaceae), a widely used traditional medicinal plant was assessed for possible sub-acute toxicity in Swiss albino rats. The rats were randomly distributed into five groups of four animals each. The groups were respectively administered 125, 250, 500 and 750 mg/kg body weight ethanolic leaf extract of Alchornea cordifolia intra peritoneally daily for (two weeks) 14 days. Normal saline was administered to the control group according to their body weights. The experimental animals were observed for another 14 days before the termination of the experiment. The weight of the animals was recorded daily throughout the duration of the study. The number of deaths in any group was recorded. All the surviving animals were sacrificed after 28 days. Blood samples were collected for biochemical and haematological analysis. Selected organs of the animals i.e., liver and kidney of both the dead and sacrificed animals were removed and stored in 10% formal saline ready for histopathological analysis. Administration of Alchornea cordifolia (0.125-0.75 g/kg, po daily) for two weeks (14 days) did not affect significantly the relative organ weights, blood chemistry and renal function. Histology of liver and kidney at dose levels up to 0.5 g/kg was normal and similarto vehicletreated controls. However, liver sections of mice treated with 0.75 g/kg Alchornea cordifolia ethanolic leaf extract showed cloudy swelling of hepatocytes with vascular degeneration. These results suggest that Alchornea cordifolia is relatively non-toxic but has the propensity to induce hepatic injury at high doses. | Journal of Analytical & Bioanalytical Techniques | 8:02:00 AMDOI: 10.4172/2155-9872.1000353
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60 | Emejulu, A.A1., Nwufo, K.C1., Ene, A.C and Obasi, U.K. (2017 | GC-FID Phytochemical Analysis and Intraperitoneal Lethal Dose (LD50) Determination of Ethanol Root Extract of Mucuna pruriens | The study was designed to evaluate the qualitative and quantitative phytochemical profile and determine the lethal dose (LD50) of ethanol root extract ofMucunapruriens. This plant is extensively used in the South Eastern Nigeria for the management of certain ailments such as Parkinson's disease, diabetes mellitus and anaemia. The quality of the phytochemical contents was determined using standard analytical methods, while a Flame ionization detector equipped Gas chromatography (GC-FID) was used for further identification and quantification of the phytochemicals. The lethal dose (LD50) was also established. Preliminary phytochemical screening revealed the presence of Alkaloids, flavonoids, tannins, saponins and phenols. Further analysis with GC-FID revealed the presence of saponins (17.6643μg/ml), sapogenin (11.8864μg/ml), catechin (4.2495μg/ml), kaempferol (3.6267μg/ml), rutin (2.5745μg/ml), anthocyanin (0.6381μg/ml), ribalinidine (73.9745μg/ml), lunamarine (2.3099μg/ml), phenols (2.7962μg/ml), tannins (5.1494μg/ml), oxalate (4.3036μg/ml), phytate (0.1891μg/ml) and spartein (0.0001μg/ml). The intraperitoneal lethal dose (LD50) value for the root extract was found to be 2154mg/kg body weight. Mucunapruriens ethanol root extract contains therapeutically useful phytochemicals with considerably safe lethal dose value. Keywords: GC-FID Phytochemical Analysis, LD50, Mucunapruriens, Intraperitoneal, root extract | International Journal of Research in Pharmacy and Biosciences | Volume 4, Issue 7, PP 23-28
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61 | Ene AC, Ezeji Chigbu NGN , Emejulu AA, Ene CU, Okwu GNand Ujowundu CO. (2018) | Antiplasmodial and Antioxidant Evaluation of the Methanol and Aqueous Extracts of SarcocephalusLatifolius | The methanol and aqueous extracts of different parts of Sarcocephalus latifolius plant were studied for their invivo anti-plasmodial activity against Plasmodium bergheiNk 65in Swiss albino mice. Phytochemicals and Antioxidantstudies were also conducted for all the plant extracts used. The aqueous extract of the stem bark of Sarcocephaluslatifolius showed the highest antiplasmodial activity.The extract significantly suppressed the parasite. There wasa significant difference (p<0.05) observed when the level of parasitemia of the animals treated with the aqueousextract of the stem bark of Sarcocephalus latifolius was compared with the untreated, chloroquine standard controland other treatment groups. Detailed phytochemical screening indicated that the crude extracts contained secondarymetabolites including phenols, tannins, alkaloids, saponins, steroids, phytate, oxalate, cyanide and anthraquinone. Theantioxidant studies showed that the methanol extract of the stem bark has significantly the highest percentage activity of 2,2-diphenyl-1-picrylhydrazyl (DPPH). The aqueous extract of Sarcocephalus latifolius yielded more of the secondarymetabolites compared to the methanol extracts. The in vitroantiplasmodial activity exhibited by these extracts mightbe attributed to these secondary metabolites. Since Sarcocephalus latifolius showed very good antiplasmodial activity,it is concluded that this plant has potentials in fighting malaria.Keywords;Antiplasmodial Activity; Sarcocephalus latifolius;Plasmodium berghei; Albino mice | SciFed Journal of Analytical Biochemistry | Volume 1 · Issue 1 · 1000005
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62 | Ezeji-Chigbu NGN, Ene AC, Emejulu AA. (2018) | Sub-Acute Toxicity Evaluation of Aqueous Stem Bark Extract of SarcocephalusLatifolius | Aqueous extract of the stembark of Sarcocephalus latifolius was administered to wistar albino rats to assess sub-acute toxicity. Twenty-four wistar albino rats were divided into four groups of six animals per group. Three groups were administered increasing doses of the extract 125, 250 and 500 mg/body weight whereas the fourth group was used as normal control. The duration of the experiment was two weeks. This study revealed that the aqueous extract of stem bark extract of Sarcocephalus latifolius affected the biochemical parameters at the test doses of 125, 250 and 500 mg/body weight. The biochemical parameters studied showed some consistent changes. There was an elevation in ALT, ALP, AST, Total Bilirubin, Direct Bilirubin, Urea, Creatinine, Bicarbonate, Chloride, Potassium, Cholesterol and Triglyceride levels in the treated groups when compared to the control groups. There were statistically significant difference (p<0.05) observed when these biochemical parameters in the treated groups were compared with the normal control. This increase in biochemical parameters was higher as the doses of the aqueous extract of the stem bark of Sarcocephalus latifolius increased. On the other hand, there was a decrease in the concentrations of Protein, Albumin,Sodium and HDL-Cholesterol in the treated groups when compared to the Control group. This decrease was observed to be statistically significant (p<0.05) when these biochemical parameters in the treated groups were compared with the normal control. There was no statistically significant difference (p>0.05) observed in the hematological parameters of the test animals compared to the control. The histopathology results of the sub-acute toxicity of the aqueous extract of the stem bark of Sarcocephalus latifolius concurs with the results of the biochemical parameters. From the result, there were no severe pathological changes in the cell architecture of the liver, kidney and heart of the extract treated animals. These results indicate that the extract may be toxic at higher doses and short term exposure. Keywords;Sarcocephalus latifolius; Aqueous Extract; Albino Rats; Sub-Acute Toxicity | SciFed Journal of Analytical Biochemistry | Volume 1 · Issue 1 · 1000002
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63 | Ibegbulem, C.O., Ene, A.C., Nwankpa, P., Chikezie, P.C. and Igwe, C.U. (2017) | Effect of wood charcoal powder on rate of microbial production of lactic acid in dehulled and undehulled Vigna unguiculata pastes | Effect of Wood Charcoal Powder (WCP) on rate of microbial production of Lactic Acid (LA) in ground dehulled and an undehulled Vigna unguiculata (cowpea) paste was studied. The pastes were analysed for proximate compositions, energy values, tannin contents and microbial loads. Later, 4.0 g pastes were treated with incremental concentrations (0.03-0.05 g) of WCP diluted with 50 mL distilled water and fermented for 2 h at ambient temperature (29.5 ± 2.0ºC). Results showed that decortication of cowpea significantly reduced (p<0.05) the crude protein, crude fibre, crude fat and tannin contents as well as energy value and microbial loads. Fermentation produced ethanol and LA. The WCP inhibited LA production in dehulled cowpea paste, whereas LA fermentation continued unhindered in Undehulled Cowpea Paste (UCP). A WCP-tannic acid-cellulose interaction study suggested that WCP interacted more with the fibre and tannin components of UCP thereby reducing the amount that would have bound the fermentative microorganisms. In conclusion, wood charcoal inhibited microbial production of lactic acid in ground dehulled cowpea paste. Keywords: Cowpea; Decortication; Lactic acid; Microbial fermentation; Wood charcoal | Journal of Nutrition and Food Sciences | 7 (2)
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64 | Chiedozie O. Ibegbulem and Paul C. Chikezie (2018) | Comparative proximate composition and cyanide content of peeled and unpeeled cassava roots processed into garri by traditional methods | ABSTRACT: Investigations on the consequential effects of the presence of cassava peelings and palm oil on the proximate composition and cyanide content during the stages of processing cassava roots into gelatinized garri meal were carried out. The peeled and unpeeled cassava roots were processed into garri and preparation of gelatinized garri meals were according to standard traditional methods. The cassava roots and garri samples were stored at ambient room temperature and 30-55% relative humidity until used for analyses. Proximate composition and cyanide contents of the samples were measured using standard methods. The moisture content of the cassava roots was significantly (p< 0.05) higher than the processed garri samples. Protein and lipid contents of the samples were relatively low, whereas the ash value of unpeeled cassava roots was significantly (p < 0.05) higher than those of the peeled samples. The carbohydrate content of the unprocessed cassava roots was comparatively lower than those of the processed roots; p< 0.05. The cyanide content of garri sample produced from unpeeled cassava roots was 4.89 folds lower than that produced from peeled cassava roots (p < 0.05). Palm oil treated garri samples gave marginal low cyanide content, which was further lowered when processed into gelatinized garri meal. The presence of cassava peelings and the addition of palm oil to cassava mash during the production of garri caused critical readjustments of samples’ physicochemical properties and impacted on the food value of the cassava-based meal.Keywords: Cassava; cyanide; garri; palm oil; proximate composition | Research Journal of Food and Nutrition | 2(2)
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65 | Chioma Y. Ezirim, Paul C. Chikezie, Kizito M. Iheanacho and Ngwu, R. Nwachukwu (2017) | Comparative Activities of Soil Enzymes from Polluted Sites in Egbema, Imo State, Nigeria. | The levels of activities of dehydrogenase, hydrogen peroxidase, catalase and lipase of different soil strata from hydrocarbon polluted site (HCPS), domestic waste polluted site (DWPS) and palm oil mill effluent polluted site (POMEPS) as well as unpolluted site were evaluated. Soil samples, namely, subsoil from 50 cm below soil surface,and topsoil were collected from different locations in Egbema urban area in Ohaji/Egbema local government area of Imo state, Nigeria. Soil enzymes activities were measured using spectrophotometric and titrimetric methods. Dehydrogenase activities of topsoil and subsoil from POMEPS were significantly (p<0.05) higher than those from unpolluted site. Comparative analysis of soil enzyme activity showed that topsoil and subsoil from HCPS gave the highest hydrogen peroxidase activity of 8.48 ± 0.01x10-3 U/g and 7.52 ± 0.02x10-3 U/g respectively. Topsoil from POMEPS gave the highest lipase activity of 1.90 ± 0.02 U/g and was followed by lipase activity of subsoil from DWPS=1.85 ± 0.02 U/g. Catalase activity of the subsoil was significantly (p<0.05) higher than that of the topsoil from all soil sites. The overall variability in enzymes activities of soil strata from different polluted sites defined the pattern of soil contamination, which could serve as biomarkers for ascertaining level of soil pollution as well as monitorial indices for bioremediation. | Journal of Pollution Effects and Control | 5(1)
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66 | Iheanacho, K.M.E., Offiah, S.A.U., Udo, M., Ugonabu, M.C., Opara, C.A., Chigbu, L., Onwubiko, D, Okechi, O.O and Igboh, N.M. (2017) | Lipid and Reductive Enzyme Profile in Hypertensive Patients | Hypertension is a major public health problem in developed and developing countries. Essential or primary hypertension is the world’s leading risk for global disease burden, and it is expected to cause more than half of the estimated 17 million deaths per year resulting from cardiovascular disease (CVD) worldwide. The objective of this study was to determine the association between serum lipid profiles and the activities of gamma glutamyl and lactate dehydrogenase in hypertensive patients and normotensive control subjects in Nigeria. This study was carried out among 60 participants of whom, 50 were hypertensive patients and ten normotensive patients. The blood pressure, lipid profile (Total cholesterol, Triglyceride, low density lipoprotein,high density lipoprotein) including enzymatic assay (gamma glutamyltransferase and lactate dehydrogenase) were determined using standard methods. Multivariate analysis of variance was used for the statistical analysis. The mean blood pressure for hypertensive were SBP and DBP were 150 and 89mmHg and 138 and 79 mm Hg for normotensive respectively. The result showed that HDL (38.48±9.457)mg/dl, was lower in hypertensive patients compared to normotensive patients with (mean ±standard deviation) of 57.00±6.06 (p>0.05).The serum level of TC, LDL, TG and LDH were higher in hypertensive patients compared to normotensive patients was significantly higher p>0.05. With increase in the TG and LDL level showed by hypertensive patients, they may be prone to cardiovascular disease compared to normotensive. Keywords: Total cholesterol, Lipo-protein, Lactate dehydrogenase(LDH), and gamma glutamyltransferase (GGT). | International Journal of Research Studies in Medical and Health Sciences. | 2 (11)
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67 | Ukairo DI, Ojiako OA, Nwaoguikpe R, Ibegbulem CO, Igwe CU and Iheme CI. (2017). | Ameliorative effects of Nigerian recipes on histopathological and immunohistochemical changes of CCl4-induced hepatic fibrosis in male Wistar rats. | The effects of methanol extracts of cooked Sphenostylis stenocarpa, Buchholzia coriacea, Chromolaena odorata and Monodora myristica on the collagen deposition of hepatic stellate cells (HSCs) in hepatic fibrotic male Wistar rats were investigated. Ninety-six adult male rats of weight 120 – 190 g were grouped into twelve (groups A-L) of eight rats each, representing normal control, olive oil, negative control, silymarin, 250 mg/kg and 500 mg/kg body weight of cooked S. stenocarpa, B. coriacea, C. odorata and M. myristica methanol extracts, respectively. Groups A and B received 1 ml of normal saline and 1 ml of olive oil, respectively, group C received CCl4/olive oil mixture only and group D was treated daily with 100 mg/kg of the standard drug, silymarin, after two weeks of CCl4 administration. Groups E - L were pre-treated with the respective extracts twice a week for four weeks by intubation before intraperitoneal intoxication with 1 ml of carbon tetrachloride (CCl4) and olive oil (1:1 v/v) mixture per kilogram body weight of the experimental animal and then extract-treatment was continued for another four weeks. Liver specimens were procured, weighed and processed for immunohistochemical studies. Necrosis and collagen deposition were evident in the liver histology of the negative control. Treatment with methanol extracts of the cooked samples, except 250 mg/kg body weight of C. odorata, ameliorated the effect of CCl4. In conclusion, methanol extracts of cooked S. stenocarpa, B. coriacea and M.myristica have anti-fibrotic and hepato-protective potentials against CCl4-induced liver injury in male Wistar rats. | Journal of Pharmacognosy and Phytochemistry | 6(6): 1624 -1631.
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