FACULTY OF BASIC MEDICAL SCIENCES
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Item PROTECTIVE EFFECT OF Pterocarpus mildbraedii HARMS EXTRACT ON PROPANIL-INDUCED HEPATOTOXICITY AND ALTERATIONS IN APOPTOTIC-RELATED PROTEINS IN WISTAR RATS(2017-05) OTUECHERE, C.AOne of the probable causes of liver diseases is exposure to environmental chemicals. Agrochemicals containing propanil are known to induce hepatic toxicity. Pterocarpus mildbraedii leaf is used in traditional medicine to treat various disorders without scientific justification. This study was designed to investigate the protective role of extract of Pterocarpus mildbraedii against propanil-induced hepatotoxicity in rats. Pterocarpus mildbraedii leaves, purchased from Oyingbo market, Lagos State, were authenticated at the University of Lagos Herbarium (LUT/5913). Powdered leaf was extracted in soxhlet, using dichloromethane:methanol (1:1), to yield crude extract of Pterocarpus mildbraedii (PME). Sixty-four male Wistar rats (130-160 g), comprising of eight groups (n=8) were used for these experiments. Rats were treated orally with normal saline (control), PME (100 mg/kg), PME (200 mg/kg), PME (400 mg/kg), propanil (200 mg/kg), PME (100 mg/kg) + propanil (200 mg/kg), PME (200 mg/kg) + propanil (200 mg/kg) and PME (400 mg/kg) + propanil (200 mg/kg) for seven consecutive days. Hepatic tissues and serum were assayed for markers of hepatic damage, oxidative stress, inflammation, and apoptosis. Aspartate aminotransferase (AST), lactate dehydrogenase (LDH), bilirubin (BIL), superoxide dismutase (SOD), catalase (CAT), reduced glutathione (GSH), malondialdehyde (MDA), myeloperoxidase (MPO) and nitric oxide (NO) were assayed by spectrophotometry. Inducible Nitric Oxide Synthase (iNOS), Cyclooxygenase-2 (COX-2), Nuclear factor kappa B (NF-κB), Caspase 3, Caspase 9, Bax, Bcl-2 expressions were measured using immunohistochemistry. Tumour suppressor p53, Bcl-2 antagonist of cell death (Bad), NF-κB, inhibitor of total nuclear factor-kappa B α (IκB α), stress activated protein kinase/ C Jun NH2-terminal kinase (SAPK/JNK), p38 mitogen-activated protein kinase (p38) and signal transducer and activator of transcription 3 (STAT 3) were assessed by ELISA. Histopathology of liver was determined by microscopy and apoptosis by TUNEL assay. Data were analysed using ANOVA at α0.05. The yield of PME was 41.9%. Administration of propanil significantly increased AST (132.10±6.32 U/L), LDH (85.70±6.60 U/L), BIL (1.15±0.16 mg/dL), SOD (0.97±0.05 U/mg protein), MDA (1.03±0.08 µgMDA/mg protein), MPO (4.98±0.12 µmol/min/mg protein) and NO (0.38 µmol/mg protein) relative to control (115.90±8.65, 32.84±9.39, 1.15±0.16, 0.38± 0.01, 0.40±0.11, 2.47±0.10 and 0.19± 0.05, respectively). Pre-treatment of propanil-exposed rats with PME (200 mg/kg) significantly decreased LDH (83%), BIL (50%), SOD (50.5%), MDA (33.1%), MPO (63.3%) and NO (59.5%). Further, propanil administration decreased the levels of GSH (2.98±0.24 µg/mg protein) and CAT (52.7±0.24 µmol H202 consumed/min/g tissue) when compared with the controls (2.04±0.09 and 51.00±0.51). However, intervention with PME restored these serum biochemical indices and antioxidant parameters back to normal values. Expressions of iNOS, COX-2, NFκB, Caspase 3, Caspase 9 and Bax were higher in the propanil group relative to control. Levels of signaling mediators p38 (81.28±7.70), STAT 3 (88.80±4.40) and NF-κB (72.76± 5.30) were lower, while SAPK (125.39±9.30), IκB α (115.83±5.60) and Bad (112.48±4.70) were higher in propanil-treated rats relative to control value set at 100. TUNEL-positive nuclei and severe periportal fibrosis were observed in tissues following propanil exposure. However, pre-treatment with PME significantly attenuated the observed propanil-induced inflammation and apoptosis. Pterocarpus mildbraedii extract protected against propanil-induced hepatotoxicity via mechanisms that involved its antioxidant, anti-inflammatory and anti-apoptotic properties.Item DETERMINANTS OF RESPONSES TO ANTIMALARIAL DRUGS IN CHILDREN WITH UNCOMPLICATED PLASMODIUM FALCIPARUM MALARIA(2011-05) SIJUADE, A.ODrug resistance is a challenge to malaria control efforts and several factors including parasite genetics, host factors and pharmacokinetics may contribute to the selection of drug resistant Plasmodium falciparum. Understanding the role of these factors in patient response to antimalarial drugs is therefore essential in the management of malaria. The aim of the study was to determine the factors contributing to delay in malaria Parasite Clearance (PC) in children and evaluating the effects of pharmacokinetic variability on treatment outcome. Children (n=2,752), aged 6 months -12 years, with falciparum malaria, were enrolled over a period of eight years and treated with standard doses of Chloroquine (CQ), Sulphadoxine-Pyrimethamine (SP) or Amodiaquine (AQ) given alone or in combination with artemisinin. Each patient was followed up for at least 14 days. Age, axillary temperature, parasite density and gametocytaemia were assessed for their potential association with delay in PC and treatment outcomes. Filter-paper blood samples were collected from some of the children (n=148) before treatment and on days 1-7, 14, 28 and 35 after treatment for determination of CQ and sulphadoxine concentrations. In another subset of patients (n=7), treated with amodiaquine, blood and saliva samples were collected over 35 days. High performance liquid chromatographic techniques were used to determine concentrations of sulphadoxine in whole blood as well as AQ and Desethyl amodiaquine (DEAQ) in saliva. Mean maximum drug concentration (Cmax), half-life (t1/2) and area under concentration-time curve (AUC0-28d) were assessed for their association and predictive value for treatment outcomes. Data were analyzed using descriptive statistics, ANOVA, Chi-square, Students’ t-test, Kruskal-Wallis and multiple regressions at p = 0.05. Age ≤ 2 years (Adjusted odds ratio [AOR] = 2.13), presence of fever (AOR = 1.33) and parasitaemia > 50,000/µl (AOR = 2.21) at enrolment were independent risk 3 factors for delay in PC, while a body temperature >38OC and parasitaemia >20,000/µl were predictors a day after treatment regardless of the drug used. Day 3 concentration ≤ 1750ng/ml and AUC0-28d ≤ 950ng/ml.h were associated with chloroquine treatment failure. In a multivariate analysis, a terminal elimination t½ ≤ 220h (AOR = 0.28) and AUC0-28d ≤ 950ng/ml.h (AOR = 4.12) were identified as independent pharmacokinetic predictors of chloroquine treatment failure. Age stratified analysis showed that SDX concentrations were significantly higher in children > 5years compared to children <5years: Cmax; 295 vs 125µg/ml, AUC0-28d; 1562 vs 812µg/ml.d. In patients who received AQ, there was a rapid conversion of AQ to DEAQ, which was detectable in plasma and saliva within 40 minutes of administration. The mean day 7 concentration of DEAQ was significantly higher in plasma than in saliva (247.8 vs 125.1ng/ml). The t1/2 of DEAQ were similar in plasma (167.25±43.4h) and saliva (146.12±17.2h). The decline phases of DEAQ in saliva concentration-time curves were approximately similar to that in plasma. Delay in parasite clearance is specific and related to drug resistance. In addition pharmacokinetic variability of sulphadoxine in children has potential impact on dosage regimen and treatment outcome.Item EVALUATION OF THE ANTIPSYCHOTIC PROPERTY OF MORIN AND ITS MECHANISMS OF ACTION IN EXPERIMENTAL MICE(2018-06) BEN-AZU, B.Psychosis is a chronic neuropsychiatric disease characterised by severe behavioural perturbations. Current drugs used in the management of the disease are associated with serious side effects. Therefore, compounds with psychotropic-antioxidant effects are currently being sought as alternatives. Morin, a naturally-occurring neuroactive flavonoid isolated from Morusalba possesses strong psychotropic and antioxidant properties, however the mechanism of the antipsychotic property has not been fully elucidated. This study was designed to investigate the antipsychotic-like activity of morin and its mechanisms of action in mice. Morin was administered intraperitoneally to male Swiss mice. Ninety mice randomised into 6 groups of each experiment (n=5): vehicle (normal saline, 10mL/kg), morin (25, 50, 100mg/kg), haloperidol (1mg/kg) and risperidone (0.5mg/kg); were pre-treated to assess the acute antipsychotic effects of morin on apomorphine-(2mg/kg), ketamine-(10mg/kg) induced stereotypes and woodblock-catalepsy test. For the chronic studies, fifty mice were given preventive treatments (n=5) with morin (100mg/kg/day), haloperidol (1mg/kg/day), risperidone (0.5mg/kg/day), or vehicle for 14 days prior to injection of ketamine (20mg/kg/day) from the 8th-14th day. For the reversal treatment, animals received ketamine for 14 days prior to the treatments. The antipsychotic and neuroinflammatory effects were also assessed in 25 mice following pretreatments with vehicle, morin, haloperidol and risperidone, in combination with lipopolysaccharide (0.1mg/kg/day) induced neuroinflammation for 14 days prior to ketamine (20mg/kg/day) treatment from the 8th-14th day. Schizophrenia-like behaviours in all chronic studies were evaluated using open-field, social-interaction and Y-maze tests. Thereafter, brain biomarkers of oxidative/nitrergic stress were determined, spectrophotometrically in specific-brain regions (striatum, prefrontal cortex and hippocampus) in preventive-reversal and neuroinflammatory studies. Specific-brain regions of dopamine, glutamate and serotonin concentrations, Glutamic Acid Decarboxylase-67 (GAD67), Brain-Derived Neurotrophic Factor (BDNF) and gp91-phox expressions were measured in the preventive-reversal study using ELISA or immunohistochemistry. Brain Tumor Necrosis Factor-alpha (TNF-α),interleukin-6 levels, cyclooxygenase-2 (COX-2) and Nuclear Factor-κB (NFκB) expressions were determined in the neuroinflammatory study using ELISA or immunohistochemistry. Dendritic arborization of the cortical pyramidal neurons of lipopolysaccharide-ketamine treated mice was assessed using silver-impregnation stain. Data were analysed using descriptive statistics and ANOVA at α0.05. Morin (25, 50, 100mg/kg) significantly suppressed stereotypy induced by apomorphine (23.4, 34.5 and 60.1%) and ketamine (33.7, 73.4 and 83.4%) relative to controls,and was devoid of extrapyramidal side effects in catalepsy test.Morin (100mg/kg) prevented and reversed ketamine-induced social and cognitive deficits relative to controls and ketamine-induced hyperlocomotion (61.6±5.2 vs109.8±5.3; 47.0±6.1 vs103.2±4.5), respectively. Morin prevented and reversed altered dopaminergic, glutamatergic, GABAergic and serotonergic neurotransmissions in the striatum, prefrontal cortex and hippocampus, respectively. Morin increased BDNF, glutathione, and decreased malondialdehyde, nitrite levels and pg91-phox expressions in the three brain regions. Morin reduced TNF-α (124.7±8.6 vs212.7±9.4; 117.3±9.7 vs278.5±13.9 pg/g tissue) in the striatum and prefrontal cortex, and morin also reduced interleukin-6 (321.3±24.2 vs704.7±26.3, 295.1±19.7 vs581.3±47.4 pg/g tissue) in the prefrontal cortex and hippocampus. It also reduced COX-2 and NFκB expressions in the three brain-regions, and increased dendritic arborization of the cortical-pyramidal neurons. Morin demonstrated antipsychotic-like activity via mechanisms related to modulation of neurotransmitters, enhancement of neurotrophin, inhibition of oxidative/nitrergic stress and neuroinflammation.Item Endocrine disruptors-arsenic, cadmium and lead in pre and postmenopausal black women with breast cancer(University College Hospital, Ibadan, 2017) Ajayi, O. O.; Charles-Davies, M. A.; Anetor, J. I.; Ademola, A. F.Background: The involvement of toxic metals in adiposity has been suggested to be contributory to the high incidence of breast cancer, particularly in sub-Saharan Africa. This study is aimed at evaluating serum arsenic, cadmium and lead in relation to adiposity and blood pressure in Nigerian women with breast cancer. Methodology: The study comprised 85 women newly diagnosed with breast cancer pre-therapy (cases) matched with 84 apparently healthy women without breast cancer (controls) according to age and menstrual phase. Arsenic (As), cadmium (Cd) and Lead (Pb) levels were determined by atomic absorption spectrophotometry. Blood pressure and anthropometry were determined by standard methods. Data analysed by Student’s t-test and Pearson correlation coefficient were considered statistically significant at p<0.05. Results: Cd and Pb levels were significantly higher in cases, compared with controls (p<0.05). Waist circumference (WC), hip circumference (HC), weight, height, waist hip ratio (WHR), waist height ratio (WHtR) were significantly higher in cases compared with controls (p<0.05). Cadmium positively correlated with diastolic blood pressure while FT4 inversely correlated with arsenic in the cases (p<0.05). Conclusion: Observations in this study suggest the involvement of these toxic metals in adiposity which could be involved in breast carcinogenesis.Item Inhibin B levels in relation to obesity measures and lipids in males with different numbers of metabolic syndrome components(2016) Laniyan, D. O.; Charles-Davies, M. A.; Fasanmade, A. A.; Olaniyi, J. A.; Oyewole, O. E.; Owolabi, M. O.; Adebusuyi, J. R.; Hassan, O.; Ajobo, B. M.; Ebesunun, M. O.; Adigun, K.; Akinlade, K. S.; Okoli, S. U.; Arinola, O. G.; Agbedana, E. O.Introduction: Defective spermatogenesis and metabolic syndrome affect 2-4% and 12.4% of males respectively. Deficient testosterone levels due to increased conversion of testosterone to oestradiol have been demonstrated in males with the metabolic syndrome (MS) with limited pituitary and leptin contribution. Defective spermatogenesis is thus implicated in males with MS but is controversial. Inhibin B is a marker of spermatogenesis. This study aims at evaluating inhibin B levels and their relationship with obesity measures and lipids in males with different number of MS components. Materials and Methods: This is a preliminary prospective study in which a total of 106 apparently healthy males (30, 30, 30 and 16 males with 0, 1, 2 and ≥3 components of metabolic syndrome (NMSC) respectively) aged 19-64 years were purposely selected. Blood pressure (BP) and obesity measures (including visceral adiposity index (VAI) and body mass index (BMI)) were obtained by standard methods. Fasting plasma glucose (FPG), total cholesterol (TC), triglycerides and high density lipoprotein cholesterol (HDLC) were determined by enzymatic methods while low density lipoprotein cholesterol (LDLC) and the lipid ratios (TG/HDLC, TC/HDLC, LDLC/HDLC) were calculated. Inhibin B was analysed by enzyme linked immunosorbent assay (RayBiotech, Inc. USA). Data analysed using analysis of variance (ANOVA) and multiple regressions were significant at P <.05. Results: Inhibin B decreased significantly in males with 0 to 2 NMSC (P <.05). However, inhibin levels between males with 0 and ≥3 NMSC were similar. Age and inhibin B levels were also similar among the different classes of BMI (P>0.05). Inhibin B related positively with HDLC and TC but negatively with VAI, LDLC and TC/HDLC. Conclusion: Reproductive function appears protected in Nigerian males with MS. However, improvement in HDLC, LDLC, TC levels, VAI and TC/HDLC may enhance fertility potential especially in males with one or two MS components, probably through dietary modulation and physical activity.Item Cytokine profile in Nigerians with tubal infertility(Termedia Publishing House Ltd, 2016) Nsonwu-Anyanwu, A. C.; Charles-Davies, M. A.; Taiwo, V. O.; Bello, F. A.; Bin, L.; Oni, A. A.Background: Immune response to genital Chlamydia trachomatis infection is involved in both immunity and pathology. The cytokine profile during infection has been implicated in the disease outcome, either resolution or severe sequelae. Serum cytokines of Chlamydia positive Nigerian women with tubal infertility were assessed to determine their possible relationship with tubal occlusion. Material and methods: One hundred and fifty age-matched consenting women (100 fertile and 50 with tubal infertility) were recruited based on C. trachomatis antibody positivity and grouped into infertile Chlamydia positive (CTpos) women (n = 50), fertile Chlamydia positive women (n = 50) and fertile Chlamydia negative (CTneg) women as controls (n = 50). High vaginal swabs and endo-cervical swabs were collected for microscopy, culture and gram staining. Cytokines [transforming growth factor β1 (TG F-β1), interferon γ (IFN-γ), tumor necrosis factor α (TNF-α), interleukin (IL )-4, IL -10 and IL-17A] were estimated by ELISA in sera. Data were analyzed using ANOVA, χ2 and Spearman’s correlation at p = 0.05. Results: Lower IFN-γ levels were observed in infertile women compared to fertile women. Fertile CTneg women had significantly higher TNF-α, and TGF-β1 compared to fertile and infertile CTpos women, respectively. Lower IL-10 levels were seen in fertile CTpos women compared to the infertile CTpos group. Vaginal discharge was negatively correlated with TNF-α and IFN-γ and positively with IL-4 in Chlamydia positive women. Conclusions: Chlamydia positive women with tubal infertility have higher IL -10 and lower IFN-γ levels than controls, which may contribute to their development of tubal pathology.Item Male sexual dysfunction, leptin, pituitary and gonadal hormones in Nigerian males with metabolic syndrome and type 2 diabetes mellitus(Avicenna Research Institute, 2016) Fabian, U. A.; Charles-Davies, M. A.; Fasanmade, A. A.; Olaniyi, J. A.; Oyewole, O. E.; Owolabi, M. O.; Owolabi, M. O.; Adebusuyi, J. R; Hassan, O. O.; Ajobo, B. M.; Ebesunun, M. O.; Adigun, K.; Akinlade, K. S.; Arinola, O. G.; Agbedana, E. O.Background: Pituitary and gonadal dysfunctions resulting from increased adiposity leading to disturbances of sexual and reproductive functions have been reported in males with metabolic syndrome (MS) and type 2 diabetes mellitus (DM2). The aim of this study was to evaluate sexual dysfunction, leptin, and reproductive hormones in Nigerian males with MS and DM2. Methods: Participants were 104 men (34 males with DM2, 17 men with MS and 53 men with normal body mass index (18.5-24.9 Kg/m2) without MS (controls)). The International Diabetes Federation (2005) criteria were used for MS diagnosis. Reproductive history, anthropometry, blood pressure (BP) and 10 ml fasting blood samples were obtained by standard methods. Fasting plasma glucose, total cholesterol, triglycerides and high density lipoprotein cholesterol were determined by enzymatic methods while low density lipoprotein cholesterol was calculated. Leptin, follicle stimulating hormone (FSH), luteinising hormone (LH), prolactin, testosterone and oestrogen were determined by enzyme immunoassay (leptin by Diagnostic Automation, Inc.; others by Immunometrics (UK) Ltd.) while oestrogen-testosterone ratio was calculated. Data analyzed using ANOVA, Chi square and multiple regression were statistically significant at p<0.05. Results: Testosterone was significantly lower in MS than controls while oestradiol and ETR were significantly higher in MS compared with controls and DM2 group (p<0.05). ETR significantly predicted testosterone in all groups (p<0.05). Significantly lower libido was observed in men in MS than controls and DM2 groups (p<0.05). Conclusion: Sexual and reproductive dysfunction may be related to increased conversion of testosterone to oestrogen in increased adipose mass in men with metabolic syndrome and type 2 diabetes mellitus.Item Reproductive function in premenopausal African blacks with metabolic syndrome: associations among Inhibin B, adipokines, pituitary and sex hormones and sex hormone binding globulin(2016) Famuyiwa, I. O.; Bitrus, D. P.; Charles-Davies, M. A.; Fabian, U. A.; Fasanmade, A. A.; Olaniyi, J. A.; Oyewole, O. E.; Owolabi, M. O.|; Adebusuyi, J. R.; Hassan, O.|; Ajobo, B. M.; Ebesunun, M. O.; Adigun, K.; Akinlade, K. S.; Okoli, S. U.; Arinola, O. G.; Agbedana, E. O.Reproductive dysfunction is associated with metabolic syndrome. Since fertility is highly valued in Africa, preliminary data on the association of metabolic syndrome with indices of reproductive function in premenopausal Nigerian women was provided. Sixty six premenopausal participants (44 with metabolic syndrome and 22 controls) aged 18-45 years were purposely selected for this study. Reproductive history, blood pressure and waist circumference were obtained by standard methods. Fasting blood was obtained for pituitary hormones, adipokines, sex hormone and sex hormone binding globulin, and inhibin B assays by EIA, ELISA and electro-chemiluminiscence. Plasma glucose, triglycerides and high density lipoprotein cholesterol were estimated by enzymatic methods. Free androgen index and oestrogen-testosterone ratio were calculated. Data obtained were statistically significant at P<0.05. All reproductive factors except follicle stimulating hormone and free androgen index levels were similar in both groups (P>0.05). Leptin levels were higher while adiponectin levels were lower in MS group than controls (P<0.05). Reproductive function appears sustained in MS. However, altered adipokines may relate to MS.Item Relationship between testosterone, oxidative stress biomarkers and antioxidant levels in male auto-mechanics in Ibadan, Nigeria(Biomedical Communications Group, Ibadan, 2016) Balogun, A. M.; Charles-Davies, M. A.; Chikezie, I. C.; Okoli, S. U.Hypogonadism attributable to males with metabolic syndrome was observed in automechanics occupationally exposed to mixed chemicals accompanied by oxidative stress (OS). We evaluated association between testosterone, OS biomarkers, enzymatic and non-enzymatic antioxidants in normal weight automechanics in Ibadan. This was a prospective cross sectional study involving 100 normal weight males aged 18 – 60 years. They were 50 automechanics in Ibadan, age and anthropometry matched with 50 eugonadic males from University College Hospital and environs (controls). Demographic, anthropometry, social habits and dietary history were obtained by standard methods. Blood (10mL) was collected and serum/plasma was used for biochemical analyses. Enzymatic antioxidants (catalase, glutathione peroxidase, superoxide dismutase (SOD) and glutathione -S- transferase (GST); non-enzymatic antioxidants (reduced glutathione (GSH), selenium and zinc), OS biomarkers (hydrogen peroxide (H2O2), malondialdehyde (MDA), total antioxidant capacity (TAC), total plasma peroxides (TPP) and oxidative Stress index (OSI) were estimated spectrophotometrically. Testosterone was assayed by enzyme immunoassay method (Dialab, Austria). Student’s t-test, Chi-square test and multiple regression were used for comparisons, associations and relationships respectively, which were significant at P<0.05. Testosterone, TPP, OSI, GST, MDA, H2O2, selenium and zinc concentrations were significantly higher while catalase and SOD concentrations were lower in automechanics than controls (P<0.05). However, testosterone levels in both groups were within the normal reference interval. TAC, OSI and GSH had significantly negative relationship while TPP had positive relationship with years at occupation in automechanics only (P<0.05). Automechanics may have OS but not hypogonadism probably due to increased antioxidant intake.Item Oxidative stress biomarkers and their relationship with testosterone in male auto mechanics in Ibadan, Nigeria(Sciencedomain International, London, 2016) Adekola, S. A.; Charles-Davies, M. A.; Onifade, A. A.; Okoli, S. U.Occupational exposure to mixed chemicals generates free radicals with inadequate antioxidants resulting in oxidative stress. Recently, hypogonadism in male auto-mechanics was associated with oxidative stress. Studies show that testosterone, a male hormone increases the activities of antioxidant enzymes. This study is aimed at evaluating the oxidative stress biomarkers and their relationship with testosterone in auto mechanics in Ibadan, Nigeria. Eighty-three males participated in this prospective cross sectional study after informed consent. Forty-three were male auto-mechanics, occupationally exposed to mixed chemicals in the mechanic community, Bodija, Ibadan (cases). Their mean (SEM) age and body mass index (BMI) were 42.5 (1.7) years and 23.8 (0.5) Kg/m2 respectively. They were age and BMI matched with 40 unexposed, apparently healthy males from the University College Hospital and environs (controls). Demography, social habits, anthropometry and gonadal status were obtained by standard methods. Serum obtained from blood (10 ml) collected from the participants was used for biochemical analyses. Testosterone levels were determined by enzyme immunoassay method (Immunometrics UK Ltd). Levels of total antioxidant capacity, total plasma peroxide (TPP), malondialdehyde (MDA), hydrogen peroxide (H2O2), glutathione peroxidase (GPX), superoxide dismutase (SOD), glutathione-S-transferase (GST), and reduced glutathione (GSH) were determined using spectrophotometric methods while oxidative stress index (OSI) was calculated. P<0.05 was regarded as significant. TPP, MDA, OSI, H2O2 and GST `levels were significantly higher (P<0.001) in eugonadal cases compared with controls. All these biomarkers levels were similar in hypogonadal compared with eugonadal cases. (P>0.05) Testosterone related negatively with SOD in the controls only but positively with MDA and negatively with GST in cases only (P<0.05). Occupationally exposed auto mechanics appear to have oxidative stress and may benefit improvement in antioxidant status. Testosterone may contribute to and enhance total antioxidant status, which may be important in gonadal function.