DSpace Community: Department of Chemical Pathology

THE RELATIONSHIP OF ENDOCRINE DISRUPTORS WITH PITUIARY GONADAL, THYROID HORMONES AND SOME RECEPTORS IN NIGERIAN WOMEN WITH BREAST CANCER

Tue, 01 Mar 2016 00:00:00 GMT

Title: THE RELATIONSHIP OF ENDOCRINE DISRUPTORS WITH PITUIARY GONADAL, THYROID HORMONES AND SOME RECEPTORS IN NIGERIAN WOMEN WITH BREAST CANCER Authors: AJAYI, O. O. Abstract: Endocrine Disruptors (EDs) such as lead, cadmium, arsenic, polychlorinated biphenyls (PCBs) and bisphenol-A (BPA) are associated with increased risk of Breast Cancer (BCa). Human epithelial receptor, oestrogen, progesterone and their receptors, gonadotrophins and thyroid hormones are also implicated in the aetiology of BCa but remains controversial. Although, several studies have been conducted on BCa, the relationship of EDs with the hormones and receptors has not been fully explored. This study was designed to examine the relationship of these endocrine disruptors with pituitary, gonadal, thyroid hormones and selected receptors in Nigerian women with breast cancer. One hundred and seventy non-pregnant women aged 48.3±1.3 years were purposively selected from the Surgical Oncology Clinic, University College Hospital, Ibadan. The study comprised of 85 Histologically-Confirmed Breast Cancer pre-therapy (HCBCa) which were sub-divided into premenopausal-HCBCa and postmenopausal-HCBCa. These were matched with 85 Apparently Healthy Women without BCa (AHWB) comprising of premenopausal-AHWB and postrnenopausal-AHWB according to age and menstrual phase. Anthropometry and Blood Pressure (BP) were determined with standard methods. Blood was obtained from participants and centrifuged to obtain serum. Serum lead, cadmium and arsenic were determined using atomic absorption spectrophotometry. Bisphenol-A and PCBs were determined using HPLC and gas chromatography, respectively. Oestradiol, progesterone, Luteinizing Hormone (LH), Follicle Stimulating Hormone (FSH), Thyroid Stimulating Hormone {TSH), Free Thyroxine (FT4) and Free Triiodothyronine (FT3) levels were determined using ELISA. Expression of Oestrogen Receptor (ER), Progesterone Receptor (PR) and Human Epithelial Receptor 2 (HER2) were determined using immunohistochernistry. Data were analysed using descriptive statistics, Student's t-test and multiple regression at a0.05 Waist Circumference (WC), Hip Circumference (HC), weight, height, Waist-Hip-Ratio (WHR), WaistHeight- Ratio (WHtR) were significantly higher in HCBCa compared with AHWB (89.9± l. l vs 82.6±1.2 cm; 101.8±1.1 vs 98.5±1.0 cm; 69.2±1.4 vs 62.1±1.1 kg; 1.63±0.0 vs 1.58±0.0 m; 0.9±0.0 vs 0.8±0.0; 55.3±0.7 vs 52.4±0.7, respectively). Toe HCBCa had significantly higher levels of lead, cadmium, arsenic, PCBs and bisphenol-A compared with AHWB (5.5±0.2 vs 1.8±0.0 μgldL; 0.04±0.0 vs 0.01±0.0 μgldL; 0.30±0.0 vs 0.04±0.0 μg/dL; 0.8±0.5 vs 0.3±0.0 μg/dL; 0.8±0.7 vs 0.4±0.0 mg/dL, respectively). The FT4 was significantly higher in HCBCa than AHWB (17.8±0.4 vs 14. 7±0.3 pmol/L). There was no difference in LH, FSH and TSH in HCBCa compared with AHWB. Progesterone and oestradiol were higher in postmenopausal-HCBCa compared with postmenopausal-AHWB. (2.09±0.35 vs 1.04±0.10 nmol/L;l56.48±12.42 vs 90.42±3.59 pmol/L, respectively) Regression analysis showed that in HCBCa, oestradiol and WC jointly predicted lead and cadmium (P>0.348, P>S.830, respectively). Diastolic-BP predicted cadmium (P=0.299), WHR and HC predicted arsenic (P>2.732) while FT3, WHtR and height positively predicted BPA (P>0.404). Contrarily, these relationships were absent in AHWB. Fifty-five tumour samples (69.6%) exhibited ER-, PR- and HER2- expressions (triple negative) while 2 (2.5%) demonstrated ER+, PR+ and HER2+ expressions. Fifty-two tumour samples (100%) in premenopausal-HCBCa exhibited ER-IPR- co-expressions while 46 (88.5%) exhibited HER2- expression. However, 18 (66.7%), 18 (66.7%) and 17 (63.0%) postmenopausal-HCBCa exhibited ER-, PR- and HER2- tumour expressions, respectively. Endocrine disruptors could have contributed to adiposity and be associated with negative receptor expression, which may induce cell proliferation and probably cause derangement in thyroid metabolism. Description: A thesis in the Department of Chemical Pathology submitted to the Faculty of Basic Medical Sciences, College of Medicine, University of Ibadan in partial fulfillment of the requirements for the award of the degree of Doctor of Philosophy, University of Ibadan, Ibadan, Nigeria.

EVALUATION OF AUTOANTIBODIES AND HEPATITIS VIRAL MARKERS IN NIGERIANS WITH LIVER DISEASES

Sat, 01 Jan 2011 00:00:00 GMT

Title: EVALUATION OF AUTOANTIBODIES AND HEPATITIS VIRAL MARKERS IN NIGERIANS WITH LIVER DISEASES Authors: OTEGBAYO, J.A.O. Abstract: Liver disease is a major public health problem. Several diagnostic and aetiologic agents of liver diseases are known, but autoantibodies to liver antigens in Nigerians with liver diseases have not been extensively studied. The study was therefore aimed at detecting the types of autoantibodies and viral antigens in various Iiver diseases among Nigerians. One hundred and twenty six consecutive patients with liver diseases and 82 apparently healthy controls were recruited at UCH lbadan. Clinical features and history of alcohol consumption were documented. Serum samples were analysed for Antinuclear Antibodies (ANA), Antimitochondrial Antibodies (AMA), perinuclear Antineutrophil Cytoplasmic Antibodies (pANCA), anti-Soluble Liver Antigen/Liver Pancreas (anti-SLA/LP) and anti-Liver-Kidney Microsomal-1 (anti-LKM-1) by enzyme linked immunosorbent assay. Similarly, hepatitis B surface antigen (HBsAg), Hepatitis B e Antigen (HBeAg), antibody to HBeAg (anti-HBe), antibody to hepatitis B core antigen (anti-HBc) and antibody to hepatitis C virus (anti-HCV) were analysed. Hepatitis B virus (HBV) DNA was determined by PCR, and samples positive for HBV s-plasmid DNA by electrophoresis were sequenced for genotypes. Liver function and prothrombin time were determined. Data were analysed using relative frequency, odds ratio, Pearson's Chi square, Fisher's exact test and Students' t-test at 5% level of significance. About 75% were males with mean age of 47.5±14.4 yrs. Sixty one percent had Hepatocellular Carcinoma (HCC), 25.4% Liver Cirrhosis (LC), 7.9% Chronic Hepatitis (Ch), 3.2% Acute Viral Hepatitis (AVH) and 1.6% Primary Biliary Cirrhosis (PBC). Hepatomegaly occurred in 78.6%, ascites in 57.1%, 51.3% consumed significant (≥80g/day for 5 years) alcohol. There was no difference in ANA among cases and controls. AMA was detectable in 60.3% of cases compared to 43.9% of controls (p<0.05). One case and one control were positive for anti-LKM-1 while all subjects were negative for anti-SLA/LP and pANCA. Anti-HBc was detected in 93.7% of cases and 73.2% controls. The prevalence of HBsAg, HBeAg anti-HBe and anti-HCV were significantly higher in cases than controls (p<0.05). The HBV-DNA was higher among cases (46%) than controls (1.2%) (odds ratio 27.3). AVH patients had the highest HBV-DNA viral load with a range of zero to 14 million copies/uL and a mean of 751.86 copies/ uL. Geometric mean HBV-DNA were 63.6, 43.15 and 7.2 copies/uL among cases with LC, HCC and CH respectively. Proportions of CH (40%) and LC (34.4%) with ANA were not significantly higher compared to controls (39.7%), but was significantly higher in HCC patients 61.9% compared to controls, The AMA was significantly higher in CH and HCC compared with controls. HBsAg was significantly higher in HCC compared to controls and other liver cases. HBeAg, anti-HBe, anti-HBc, anti-HCV and HBV-DNA were significantly higher in CH, LC and HCC compared to controls (p-0.05). There were 53 genotype E and two genotype A in cases while only one control had genotype E. Prevalence of autoantibodies to liver antigens is similar in individuals with or without liver diseases and, therefore not reliable in predicting autoimmune liver disease. HBsAg, anti-HBcAg, anti-HBV and HBV-DNA were strongly associated with liver disease. HBV genotype E is predominant in Nigeria. Description: A Thesis in the Department of Chemical Pathology, submitted to the Faculty of Basic Medical Sciences, College of Medicine, in partial fulfillment of the requirements for the Degree of Doctor of Philosophy of the University of Ibadan, Ibadan, Nigeria.

THE ROLE OF OXIDATIVE STRESS IN BIOCHEMICAL, SEMINOLOGICAL AND HISTOLOGICAL CHANGES DURING ACUTE ADMINISTRATION OF ARTEMETHER

Fri, 01 Jan 2010 00:00:00 GMT

Title: THE ROLE OF OXIDATIVE STRESS IN BIOCHEMICAL, SEMINOLOGICAL AND HISTOLOGICAL CHANGES DURING ACUTE ADMINISTRATION OF ARTEMETHER Authors: ADEKUNLE, A. S. Abstract: Artemether is an antimalarial drug which is effective in the treatment of multidrug resistant malaria but also induces oxidative stress in tissues. Limited information is available on role of oxidative stress in tissue toxicity during administration of artemether. Therefore biochemical, seminological and histological changes following acute administration of artemether were evaluated. One hundred and sixty male albino mice weighing 35-37g were divided into six groups and used for the animal study. Groups 1, 2 and 3 consisted of 30 uninfected mice each given 1.2mg/kg, 24mg/kg and 4.8mg/kg artemether respectively. Group 4 consisted of 20 Plasmodium yoelli infected mice, treated with 1.2mg/kg artemether: group 5 consisted of 20 P. yoelli infected and untreated mice, while group 6 (control) consisted of 30 uninfected and untreated mice. Artemether was administered to groups 1-4 for five consecutive days after which the mice were sacrificed on day 6. Hepatotoxicity was assessed by assay of liver enzymes, lipids and lipoproteins, total proteins, catalase, superoxide dismutase, reduced glutathione activities, malondialdehyde and histological examination of liver. Effects on brain and kidney were assessed by histological examinations of the organs. Tissue homogenates of brain, liver, kidney and testes were used for the assay of malondialdehycle and antioxidants. Effect of artemether on male fertility was assessed by sperm analysis, testicular weight, testicular testosterone and histological examination of testes. In the human study, effects of artemether on liver in 20 male patients suffering from malaria disease and 20 apparently healthy male individuals (age matched) were determined by measuring changes in serum gamma glutamyl transferase, alkaline phosphatase, alanine aminotransferase, albumin, total proteins, total cholesterol, triglyceride and lipoproteins. The dosage was 200mg on day 1 and 100mg on days 2-5. Testosterone was analysed by ELISA while other biochemical parameters were analysed using spectrophotometric methods. Sperm analyses were done using the improved Neubeur's counter. Student's t- test and ANOVA were used for data analysis. There was increased lipid peroxidation in the four organs as indicated by increased malondialdehyde and antioxidants (reduced gIutathione catalase and superoxide dismutase) in groups given artemether when compared with control. In artemether treated groups, there were dose-dependent histopathological effects which include degeneration of interstitial tissues in testes and significant reductions in sperm counts, motility and viability. Activities of alkaline phosphatase in groups 1 (658.63±81.1iu/l) and 4 (666.80 ±83.2iu/l) were significantly higher than control (407.60±63.7iu/l) (p<0.05). In group 4, there were significant differences in total cholesterol (81.53±18.9mg/dI vs 49.80±3.2mg/dl), triglyceride (95.58±21.1mg/dl vs 50.00±3.5mg/dl), total proteins (7.10±0.3mg/dl vs 6.00±0.3mg/dl), catalase (2.48±1.1 vs 4.00±1.2unit/g) and superoxide dismutase (1.20±0.5 vs 1.57±1.2unit/g) when compared with control (p<0.05). There was no significant difference in the biochemical parameters in patients suffering from malaria after administration of artemether when compared with baseline. Apparently healthy volunteers showed significant changes in total protein (6.77 ±1.5 vs 7.00±1.73g/dl), albumin (3.24±0.71 vs 3.85±1.32g/dl), and gamma glutamyl transferase (51.17±2.13 vs 42.53±1.70iu/l) after administration of artemether when compared with baseline (p<0.05). Artemether induced oxidative stress which resulted in reduced sperm quality and defective histology of the animal tissues. Artemether did not cause any significant adverse effects in humans. Description: A Thesis in the Department of Chemical Pathology submitted to the Faculty of Basic Medical Sciences, in partial fulfillment of the Degree of Doctor of Philosophy of the University of Ibadan, Nigeria.

EFFECT OF CO-ADMINISTRATION OF ACETAMINOPHEN AND METHIONINE ON MICRONUTRIENT METABOLISM

Sat, 01 Jun 2013 00:00:00 GMT

Title: EFFECT OF CO-ADMINISTRATION OF ACETAMINOPHEN AND METHIONINE ON MICRONUTRIENT METABOLISM Authors: IYANDA, A. A. Abstract: Generally, altered micro-nutrient levels affect the anti-oxidant defense mechanism and impact significantly on genomic stability. Acetaminophen induces tissue damage, which is prevented by methionine. However, the influence of acetaminophen and methionine interaction on micro-nutrient levels is unknown. In this study, the possible acetaminophen and methionine-induced micro-nutrient alterations were investigated. One hundred and sixty female Wistar rats (270-350g) divided into 20 groups of eight rats each were administered with acetaminophen, and acetaminophen and methionine (p.o) for the acute and chronic studies. In the acute study, 350, 1000, 3000 and 5000 mg/kg body weight of acetaminophen were administered to groups 1-4 respectively, as well as 5-8. Similar doses of acetaminophen and methionine (5:1) were administered to groups 9-12 and 13-16 while normal saline was administered to group 17 (actute-control). In the study, groups 18 and 19 were administered daily with 350 mg/kg acetaminophen and acetaminophen and methionine respectively while group 20 (chronic-control) received normal saline and were sacrificed after 30 days. Groups 1-4 and 9-12 rats were sacrificed at 4 hours (peak of absorption) while those in groups 5-8 and 13-16 were sacrificed at 16 hours (peak of toxicity). Sera obtained from blood (8 mL) were analysed for Superoxide Dismutase (SOD), liver markers (alanine aminotransferase-ALT, aspartate aminotransferase-AST, alkaline phosphatase-ALP, gamma-glumly transferase, bilirubin, total protein, albumin) and renal function markers (urea, creatinine) by spectrophotometry. Micronutrients- Cu, Cr, Fe, Se, Zn were determined by atomic absorption spectrophotometry while high-performance liquid chromatography was used for the determination of vitamins (A, C, E) and folic acid. Hepatic and renal tissues were subjected to histological assessment. Data were analysed using Pearson's correlation coefficient, Student's t-test and analysis of variance. Level of significance at p=0.05. Acetaminophen and methionine-treated rats showed similar levels of hepatic and renal function markers as control in both acute and chronic studies. However, at 16 hours, acetaminophen-treated rats (1000 mg/kg) had respectively significantly higher levels of AST, ALT and urea (487.3±80.4 IU/L, 379.7±153.9 IU/L; 10.6±1.8 mmol/L) compared with acetaminophen and methionine-treated rats (33.8±12.1 IU/L, 3.40±0.7 nmol/L) and control (36.4±10.8 IU/L, 33.8±12.1 IU/L, 3.40±0.7 mmol/L). Renal histology revealed tubular necrosis (acetaminophen) and no visible lesion (control; acetaminophen and methionine). Comparison of micronutrients in acetaminophen and methionine treated rats between each of the doses and controls at 4 hours showed significant differences except Cu at 1000 and 5000 mg/kg (134.1±3.9 and 147.1±29.3 ug/dL), vs control (139.9±8.5 ug/dL). Fe at 3000 mg/kg control (133.2+21.6 vs 145.7±5.2 ug/dL); folic acid at 350 mg/kg vs control (8.4±1.3 vs 8.1± 0.1ng/mL) and vitamin E at 350mg/kg vs control (0.9±0.1 vs 0.9±0.1 mg/dL). At 16 hours, all micronutrients except Cr at 3000 (0.2±0.1, 0.2±0.1 ug/L) mg/kg had significant alterations. Significant reduction in activity of SOD was recorded at 1000, 3000 and 5000mg/kg (11.9±0.1, 11.3±0.1 10.8±0.1 Umg/protein) vs control (12.8±0.3 Umg/protein). Co.administration of acetaminophen and methionine induced alterations in micronutrient levels in nephroprotected rats. This may be attributed to increased free radical generation suggestive of oxidative stress. Supplementation with micronutrients may be necessary during acetaminophen and methionine drug administration. Description: A Thesis in the Department of Chemical Pathology, submitted to the faculty of Basic Medical Sciences, College of Medicine in partial fulfillment of the requirements for the degree of Doctor of Philosophy of the University of Ibadan, Nigeria.