Medical Biochemistry

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Assessment of Urine Cotinine Levels in Active and Passive Smokers, and Studies of Cardiovascular Risk Factors in Ogolcho Town, Oromia, Ethiopia
(Addis Ababa University, 2015-06) Dedefo, Gobena; Ashall, Frank (PhD)
Introduction: Tobacco use is a global escalating public health problem, especially in low-income countries, one-third of the adult males. Non-communicable diseases (NCDs) are a growing problem in Africa and it is partially caused by tobacco use. There are few studies of smoking practices and other cardiovascular risk factors in Ethiopia and it is unclear how prevalent many of these problems are, because their prevalence varies in different areas of the country. A more recently survey of smoking prevalence in 187 countries showed that global smoking prevalence fell from 41% in 1980 to 31% in 2012, and that Ethiopia ranks in the bottom five countries with the lowest smoking prevalence (below 10%), but there is evidence that smoking prevalence is increasing in Ethiopia. Objective: To assess active tobacco and passive smoking, for further screening of major cardiovascular risk factors (serum cholesterol levels, hypertension, fasting blood glucose, Body Mass Index and waist circumference) among residents of Ogolcho town, Oromia; and to examine carbon monoxide levels in homes in Ogolcho town. Methods:Across-sectional study was conducted on tobacco users, passive smokers and non-smokers in a rural population of Ethiopia in Ogolcho, from May 2014– May 2015.Subjects were interviewed, using a standardized WHO questionnaire, about their lifestyle habits and substance use. Blood specimens from individuals over 18 years old were taken for serum lipid profile and blood glucose levels. Urine was screened for cotinine (a nicotine metabolite) in active and passive smokers, using a One Step cotinine test that detects urine cotinine levels above 200 ng/mL.Anthropometric indices (BMI, waist circumference) and blood pressure were measured. Frequencies ofthe different variables were analyzed statistically. Results: Of 95 total participants, 33% (31) were active smokers, 56.3% (54) were passive smokers, 8.6% (8) were neither active nor passive smokers, and 2% (2) were purely shisha smokers. Over 80% of active and passive smokers chewed khat regularly, whereas about a half of smokers and non-smokers used alcohol. All active smokers smoked either in their house or in public places, where exposure of many nonsmokers, especially children and women, occurs. A high percentage of smokers smoked at home (41%), and passive smokers that were strongly positive for urine cotinine included two children aged 4 and 5. Of 85 passive and active smokers, 67 % (57) were positive for urine cotinine. Almost all (90%) active smokers, and 54% of passive smokers, were positive for urine cotinine. Cardiovascular risk factors other than smoking were common in the population studied. Active smokers had 25.8% prevalence of abdominal obesity, 48.4% were underweight, 3.2% overweight, 54.8% had elevated triglycerides, 42% were hypertensive, 48.4% had low HDL and 3.2% high fasting glucose. 10.3% of passive smokers had abdominal obesity, 17% were underweight, 24% overweight, 55% hypertensive, 38 % had elevated triglycerides and 24% had low HDL. Prevalence of CVD risk factors among non-smokers were: 12.5% elevated blood pressure, 50% abdominal obesity, 12.5% overweight and 37.5% overweigh. Non- smoker lipid profile prevalence was: elevated total cholesterol in 37.5% of subjects and decreased HDL level prevalence in 25% of non-smokers. Carbon monoxide levels were generally safe in homes, except within centimeter distances of charcoal stoves. Conclusion: This cross-sectional analysis indicates that khat and alcohol consumption, or both combined, are common among smokers and non-smokers in Ogolcho, and the prevalence of passive smoking is high. Urine cotinine testing was an effective way of studying passive and active exposure to cigarette smoke. The prevalence of CVD risk factors was high in the studied population, strongly supporting the importance of screening, prevention and treatment of smoking and other cardiovascular risk factors in Ethiopia.
Effect of Ginger Rhizome Homogenate, Ginger Rhizome Steam Distillate, and Simvastatin on Fasting Blood Glucose and Serum Lipid Profiles of Streptozotocin-induced Diabetic Balb/c Mice
(Addis Ababa University, 2015-07) Mulu, Aweke; Ashall, Frank (PhD)
Introduction: Diabetes is an increasing problem in Ethiopia, affecting up to 6.5% of Ethiopian adults. There are serious complications associated with diabetes, particularly macrovascular and microvascular complications. Controlling blood glucose significantly reduces the risk of complications of diabetes. Insulin therapy is the only current treatment for type 1 diabetes. Type 2 diabetes can be treated with insulin as well numerous oral and injectable medications. However, current diabetes treatment has disadvantages, such as drug-related hypoglycemia and high cost. Traditional medicine is used to treat diabetes across the world and is readily available and inexpensive. Ginger (Zingiber officinale) is widely consumed as a spice, and numerous studies suggest that ginger may have beneficial effects for diabetes and dyslipidemia. However, further studies are needed to evaluate these effects of ginger. Objectives: To evaluate the effects of homogenized ginger rhizome, and a steam distillate of ginger rhizome containing 25% gingerol, on blood glucose and fasting lipid profiles of streptozotocin (STZ)-induced diabetic mice and to compare these effects with those of simvastatin. Methods: Balb/c mice were randomly assigned to receive ginger rhizome homogenate, steam distillate of ginger rhizome or simvastatin treatment. Two doses of streptozotocin (33mg’kg) were given intraperitoneally a week apart to induce diabetes. Mice were treated with 2g/kg/day of ginger rhizome homogenate by oral gavage, or about 3 mL/kg/day of steam distillate of ginger rhizome by oral gavage, or 40mg/kg/day simvastatin intraperitoneally. Body weights, blood glucose and lipid profiles were measured. Results: Oral gavage with ginger rhizome homogenate caused significant blood glucose increases, whereas a steam distillate of ginger rhizome showed significant blood glucose lowering effects, in STZ-induced diabetic mice. Diabetic mice treated with ginger rhizome homogenate for 56 days showed significantly lowered serum LDL and triglyceride, and higher HDL levels compared with non-treated diabetic mice but had no significant effect on total cholesterol. The steam distillate of ginger rhizome, given by oral gavage, lowered LDL but had no significant effect on HDL, total cholesterol or triglyceride levels of STZ-diabetic mice. Simvastatin (40mg/kg) given intraperitoneally decreased blood glucose, decreased LDL and increased HDL, but had no significant effect on triglycerides and total cholesterol in STZ-diabetic mice. None of these treatments, ginger rhizome homogenate, steam distillate, or simvastatin, had a major effect on weights of diabetic mice. Conclusions: Ginger rhizome homogenate treatment exacerbated hyperglycemia in STZ-diabetic mice, but improved diabetic dyslipidemia, while a steam distillate of ginger rhizome and simvastatin each decreased fasting blood glucose and improved lipid profiles of STZ-induced diabetic mice.
Effect of resveratrol, Metformin and Eucalyptus Oil on Visceral Fat Deposition, Serum glucose, Lipid Profiles and Liver Function Tests of Swiss Albino Mice Fed a High-fat diet
(Addis Ababa University, 2016-02) Matheos, Tigist; Ashall, Frank (PhD)
Introduction: Obesity results from environmental and genetic factors and is rapidly growing public health problem with its leading co-morbidity type 2 diabetes. Great emphasis has been put on establishing treatments for obesity, including pharmacological intervention using animal models of obesity. Objective: To develop an obesity model in Swiss albino mice using a high fat diet, that is feasible and affordable in Ethiopia, and to assess the effect of resveratrol, metformin and Eucalyptus oil on body weights, visceral fat deposition liver histology, serum glucose, lipid profiles and liver function tests in these mice. Methods: Male Swiss albino mice were fed a normal chow diet, high fat diet (52% fat, w/w) containing lard made from choma (crude beef fat), a high fat diet with resveratrol (75 μg/ml) in the drinking water for 12 weeks, a high fat diet with intraperitoneal metformin injections (250 mg/kg/day) and a high fat diet with Eucalyptus oil (10μL/mouse/day) administered intraperitoneally for 8 weeks. Body weight, visceral fat mass, blood glucose, serum lipid profiles, serum liver enzymes, and liver histopathology were assessed. . Results: The high fat diet did not cause significant changes in total body weight of mice, but there was significantly more visceral fat deposition as large fat pads in mice fed a high fat diet for 12 weeks (1140.0 ± 248.8 mg, as compared with 66.5 ± 27.6 mg in normal diet fed mice) and also in mice fed a high fat diet for 8 weeks (862.4±148.7 mg, as compared with 611.7 ±178.3 mg for mice fed a normal diet). The high fat diet did not induce any obvious prediabetic or diabetic state and did not raise fasting blood glucose level above the normal range. Resveratrol administration to Swiss albino mice fed a high fat diet did not cause any significant effect on body weight, visceral fat mass, fasting blood glucose, liver enzymes, or serum lipid profile. Treatment with metformin did not have any significant effect on body weight, fasting blood glucose level, visceral fat mass, serum liver enzymes, serum total cholesterol level or serum triglyceride level, but metformin significantly increased serum HDL level, and decreased serum LDL level, and raised serum AST level. Eucalyptus oil did not have any significant effect on body weight and visceral fat mass, serum ALT level, serum ALP level, serum total cholesterol and on serum triglyceride level, but it significantly lowered serum LDL levels and raised serum HDL level, as well as weakly increasing serum AST. Eucalyptus oil also lowered blood glucose levels even though they were within the normal range to begin with. Conclusion: The high-fat lard diet developed in a Swiss albino mouse model may be a useful model for visceral obesity, but, without modification, was not a model for prediabetes or diabetes. Resveratrol had no effect on body weight, fat deposition and metabolic profiles. Metformin caused elevation of HDL and lowering of serum LDL. Eucalyptus oil lowered fasting blood glucose and serum LDL levels and it elevated serum HDL levels. The significance of these findings are discussed. Keywords: Obesity, High fat diet, AMPK, Resveratrol, Metformin, Eucalyptus oil
The Effects of Coriandrum Sativum Seed Extracts on Hyperglycemia, Lipid Profile and Renal Function in Streptozotocin Induced Type - 2 Diabetic Swiss Albino Mice
(Addis Ababa University, 2014-05) Yibru, Endris; Ashall, Frank (PhD)
Background: Convectional drug treatment for diabetes mellitus carries risks that lead to many adverse effects such weight loss, hypoglycemia and many others. Ethiopia is rich in natural resources and medicinal plants useful in the treatment of diabetes. Aim of the study: To investigate the effect of Coriandrum sativum seed extracts on hyperglycemia, lipid and renal profile in streptozotocin induced diabetic mice. Methods: Thirty six male Swiss albino mice were kept in six different groups for 21 days. Group I served as normal controls; Group II served as diabetic control; Groups III, IV and V were given 300 mg/kg, 400 mg/kg and 500mg/kg of Coriandrum sativum seed extracts (70% ethanol), respectively; and Group VI received 5mg/kg glibenclamide drug. The effect of extracts on hyperglycemia, lipid profile and renal function were tested by chemistry analyzer. Results were analyzed using one way ANOVA at a 5% level of significance. Results: The fasting blood glucose level was significantly (p<0.05) reduced at 400mg/kg and 500mg/kg of Coriandrum sativum extract concentration as compared to the diabetic group. It also reduces total cholesterol, triglyceride, low density lipoprotein, urea and creatinine, and improves high density lipoprotein level and total protein in treated diabetic mice. Conclusion: Reduction in the fasting blood glucose, total cholesterol, triglyceride level, low density lipoprotein, urea, creatinine, and improvement in the high density lipoprotein and total protein by Coriandrum sativum extract indicates that it has anti-hyperglycemic, anti-hyperlipidemia and renal failure restoration effect in streptozotocin induced diabetic mice. Keywords: Diabetes mellitus, Coriandrum sativum, Hyperglycemia, hyperlipidemia, Renal function
Investigation of Toxicity of Cigarette Butts Collected in Addis Ababa to Swiss Albino Mice
(Addis Ababa University, 2016-01) Tefera, Tigist; Ashall, Frank (PhD)
Background: Cigarette butts are the most common form of litter in the world; approximately 5.6 trillion cigarettes are smoked yearly worldwide. Over 7000 chemicals may be introduced to the environment in cigarette butts and smoke constituents. These include chemicals such as nicotine, carbon monoxide, hydrogen cyanide, polycyclic aromatic hydrocarbons, heavy metal ions, nitrosamines, ammonia, acetaldehyde, formaldehyde, benzene, phenol and acetone, over 70 of which are carcinogenic to humans. Environmental cigarette butts are toxic to microorganisms, fish, other marine organisms and birds, but no studies have been done on their toxicity to mammals. Methods: In this study the toxicity of cigarette butt leachates to Swiss albino mice was investigated, and the effects of cigarette butts with and without associated tobacco were compared. Cigarette butt leachates were made by soaking different numbers of cigarette butts in 250 mL of tap water overnight, and leachates were given to mice to drink. The weights of mice as well as amounts of food and fluids consumed were followed over time. Necropsies were performed and tissues were weighed and evaluated by histological staining with eosin and hematoxylin. Blood glucose and liver function tests (ALT, AST and ALP) were also measured. Results: Mice given high-concentration cigarette butt leachates during their growth from 6 weeks to 16 weeks preferred water over any of the leachates, whereas mice given lower butt concentrations of leachates representative of the relative frequencies of butt types in the environment drank butt leachates as well as water. Food intake was not affected by cigarette butt leachates. Mean fasting blood glucose and activities of serum liver enzymes were not different between mice given leachates or water to drink. The weights of the mice, as well as tissues (liver, lung, spleen, heart, kidney, epididymal fat pads) were significantly lower in mice that drank leachates with associated tobacco than in mice that drank water alone or leachates made from butts without associated tobacco. This suggests that a component(s) of tobacco is/ are responsible for reducing weight gain during mouse growth. Cigarette butt leachates had no effect on histopathology of liver, heart and kidney, whereas lung tissue of the mice that drank leachates made with tobacco- associated butts showed increased air space volumes and alveolar fibrosis, compatible with pulmonary emphysema. Conclusions: Cigarette butts are toxic to laboratory mice, causing reduced weight gain during growth, reduced mass and size of tissues and organs, and pulmonary emphysematous changes. An average cigarette butt thrown into the environment contains 1.5 mg of nicotine, enough to potentially kill 15 mice. The implications of these studies to environmental animals in Ethiopia and elsewhere are discussed
Investigation of Urine Cotinine Levels as Indicators of Tobacco Use and Passive Smoking, with Parameters of Metabolic Syndrome, Among Adults in Chena Wacha
(Addis Ababa University, 2015-03) Wondimu, Anteneh; Ashall, Frank (PhD)
Background: Active and passive cigarette smokers produce the nicotine metabolite, cotinine, which can be detected in the urine. Smoking increases the risk of metabolic syndrome, includes abnormalities in lipid profile, blood glucose, blood pressure, BMI and waist circumference. Objective: The study, investigate urine cotinine levels as indicators of tobacco use and passive smoking, with parameters of metabolic syndrome among adult in Chenna wacha, SNNPR. Method: A cross sectional study was done on a total of seventy smokers (46 active and 24 passive). Smoking history was obtained from the participants which was then used to calculate smoking consumption in pack-years. In this study, current active smokers were classified based on the number of pack-years as into mild to moderate (1 – 10 pack-years) and heavy (more than 10 pack-years. Parameters of metabolic syndrome were determined using standard methods, while urine cotinine was determined using COT® one step cotinine test device at a cut-off of 200 ng/mL. Data analyses were performed with SPSS16.0 system. Result: The finding of the study indicates that 88.7 % of active smokers and 11.3 % of passive smokers were positive for urine cotinine, with substantial agreement between cotinine test results and self-reported cigarette use (kappa 0.63). Among five children taken from four families in which there was an active smoker in the home, two out of the five (40%) were positive for urine cotinine. The study also shows that the prevalence of metabolic syndrome, based on the occurrence of at least three components of metabolic syndrome, was 5/46 (10.8 %) in active smokers and 2/19 (10.5 %) among passive smokers. Taking mild to moderate smokers as reference, heavy smokers were relatively underweight with statistical significance (OR 7.90; p= 0.004). Adult passive smokers were not significantly underweight compared with mild-to- moderate smokers (OR 2.09; p= 0.234). As the number of pack-years of smoking increased among smokers, they had an increased number of parameters of metabolic syndrome. Conclusion: This study shows that urine cotinine measurements are a practically convenient and valuable tool for objectively assessing smoking and passive smoking in Ethiopia, and detected passive smokers, including children as young as 6 years old. Some parameters of metabolic syndrome showed a significant association with the level of smoking. Key words: Cotinine, urine, metabolic syndrome
Screening of Parameters of Metabolic Syndrome and C-Reactive Protein (CRP) in Cigarette Smokers in Adama, Ethiopia
(Addis Ababa University, 2014-01) Teshome, Abinet; Ashall, Frank (PhD)
Background – Several previous studies indicated conflicting findings on parameters of Metabolic Syndrome and CRP in cigarette smokers. The aim of this study was to determine whether there is difference in levels of parameters of Metabolic Syndrome and CRP in cigarette smokers and non- smokers in Adama, Ethiopia. Methods – subjects were 99 healthy male individuals from age 15 to 60 years. The subjects were divided into two groups of 50 smokers and 49 non- smokers. BMI, blood pressure plasma levels of total cholesterol, triglycerides, HDL, and CRP were measured and LDL cholesterol was estimated using the Friedewald formula. Results – Overall subjects’ plasma levels of total cholesterol, triglyceride, LDL, HDL and serum CRP were significantly elevated in smokers. No significant difference was found in BMI, systolic and diastolic blood pressure between cigarette smokers and non-smokers. In smokers alcohol drinking was significantly correlated with increased plasma HDL, systolic and diastolic blood pressure while khat chewing was correlated with increased plasma total cholesterol. Conclusions – The results showed no difference in BMI between smokers and non smokers and also there was no significant difference in blood pressure in smokers and non smokers. Cigarette smoking is correlated with increased total cholesterol, TG, LDL, HDL and CRP. Among smokers blood pressure, total cholesterol and HDL were significantly higher in those whose alcohol consumption was higher.

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