2. Objectives 1.To evaluate the clinical utility of biochemical tests, ie. a. Alpha-Fetoprotein (AFP), b.Alkaline Phosphatase (ALP), c.Gamma-glutamylase transferase (GGT) d.isoenzymes of ALP and GGT in the diagnosis of PHC 2.To determine the seroprevalence of HBV and HCV in PHC and HBV in HIV 3.To establish some aetiological factors associated with PHC

3. Methods LFT: RA1000 Technicon, ALP and GGT isoenzymes: PAGE in Bio-Rad Dual Slab Gel and LKB laser densitometer AFP:RIA I 125 kit HBsAg: Bioelisa Biokit; Anti HCV: MUREX kit ABBOTT and substrate TMB Fibro Test: Alpha-2-macroglobulin, haptoglobin, Apo A-1, Total Bilirubin and GGT on Beckman Coulter AU680 Chemistry Analyser Calculated on www.biopredictive.comwww.biopredictive.com FIB-4 index calculation: (age in years X AST IU/L X Platelets X 10 9 )/L)/(ALT IU/L).

4. Methods 7 Cross-Sectional Studies, 1998-2015 Sites:  Parirenyatwa & Harare Hospitals, Opportunistic Infection Clinics  National Blood Transfusion Services, Zimbabwe 571 Patients 18-55 years, healthy controls: 331

5. Methods The liver function tests and alpha-fetoprotein were determined as in method section in 160 patients with a clinical diagnosis of PHC. Data was interpreted using Biochemical Reference Values in Zimbabwe (Marima- Matarira 1989) except for Alpha-fetoprotein.

6. Methods The haematological and biochemical investigations were carried out on 160 patients with a clinical diagnosis of PHC. The methods are as described in the method section. Data from patients was interpreted using the reference values of healthy donors in Zimbabwe (Marima-Matarira 1989)

7. Table 1: Abnormal Liver Function Results and Alpha-Fetoprotein in PHC TestCut-Off PointPercentage 1.Aspartate Aminotransferase 2.Alkaline phosphatase 3.Gamma-glutamyl Transferase 4.Total Bilirubin 5.Conjugated Bilirubin 6.Alanine Aminotransferase 7.Alpha-fetoprotein 8.Albumin 9.Cholesterol (random) 10. Total Protein > 50 IU/l > 120 IU/l > 40 IU/l > 40 umol/l > 14 umol/l > 50 IU/l > 400 ng/ML < 30 g/l > 8.5 mmol/l < 2.4 mmol/L > 84 g/l < 54 g/l 83% 81% 75% 50% 48% 47% 10% 9% 7% 6%

8. Males : Female Ratio4:1 Mean ages of 3 studies in years56, 46, 43.8 declining Patients who took alcohol : abstainers6:1 Patients who admitted to heavy alcohol use 52% Mean survival period5.3 months Mean symptoms before hospital Presentation 5.2 PATIENTS DATA AND SOCIAL HISTORY

9. Table 2: HAEMATOLOGICAL,BIOCHEMICAL INVESTIGATIONS IN PHC [ n=85] TESTCUT-OFF POINTPERCENTAGE 1. Haemoglobin< 12.5 g/l47% 2. RBS< 4.2 x 10 12 / 1 47% 3. WBC> 10.5 x 10 9 / 1 26% 4. Urea> 6.0 mmol/l23% 5. Creatinine> 190 µmol/l23% 6. Sodium< 126 mmol/l18% 7. Chloride< 99 mmol/l 7% 8. Potassium> 5.6 mmol 1% 9. Albumin< 30g/l10%

10. TABLE 4: THE HISTOLOGICAL ANALYSIS OF LIVER TISSUE OF PATIENTS WITH A CLINICAL DIAGNOSIS OF PHC PATHOLOGYPERCENTAGE OF TOTAL PHC 47 PHC and Cirrhosis 12 PHC and Haemochromatosis 2 PHC, Cirrhosis and Haemochromatosis 2 Secondary Hepatocellular Carcinoma 2 Cirrhosis only 12 Cirrhosis and Haemochromatosis 1 Haemachromatosis only 7 Others 11 Normal 4 Total100

11. Table 4 continued:- Others included: malaria,schistomiosis, adenocarcinoma, apudoma, drug induced cholestasis, fibrotic gall bladder, lymphoid tissue tumour cells, cholestasis, bile thrombi and chronic cholecystitis. In the PHC patients with cirrhosis, 70% had micronodular while 30% had macronodular

12. Table 6: ALKALINE PHOSPHATASE AND GGT ISOENZYMES IN HEALTHY CONTROLS, PHC PATIENTS PatientsAnodal (%)Cathodal (%) Healthy Controls (100)(3.9) PHC (100)83.3 p < 0.05 GGT Isoenzyme 1 Abnormal96 p < 0.05 Sera from healthy controls, patients with PHC was run on PAGE electrophoresis. A highly specific substrate for ALP, p-touluidinium-5-bromo-4 chloro-3-indoyl phosphate was used to stain the isoenzyme at room temperature (17˚C). For GGT trichloroacetic acid and glycerol gave lilac bands at 17˚C.

13. SEROPOSITIVITY FOR HBV AND HCV IN PHC AND HEALTH BLOOD DONORS Test PHC (%) (n=60) Controls (%) (n=30) Anti HCV positive20 0 HBsAg positive48.313.3 Dual Anti-HCV and 8.3 0 HBs Ag positive Anti- HCV and or6013.3 HBsAg positive

14. THE DISTRIBUTION OF STAGES OF LIVER FIBROSIS IN HIV POSITIVE on HAART AND HIV NEGATIVE PARTICIPANTS

15. THE PREVALENCE OF CLINICALLY SIGNIFICANT FIBROSIS IN HIV POSITIVE on HAART AND HIV NEGATIVE PATIENTS HIV negative (n=32) HIV positive (n = 79) Total (n = 111) AST/ALT ratio N (%) 84.4%79.7% (Over estimates) 81.1% APRI test N (%) 6.3% 13.9% (Under estimates) 11.7% FIB-4 index N (%) 9.4% 22.8% 18.9% The prevalence of clinically significant fibrosis in HIV positive patients as determined by the AST/ALT ratio APRI test and FIB-4 index were 79.7%, 13.9% and 22.8% respectively.

16. HBV CO-INFECTION IN HIV POSITIVE PATIENTS

17. References Central African Journal of Medicine, (1985)31 No 11:211-214. Central African Journal of Medicine,(1985) 31 No 5:98-100. Central African Journal of Medicine, (1988) 34. No. 7: 169-170. Annals of Clinical Biochemistry,(1991) 28:512-513. Afro-Arab Liver Journal, 1: (1994)69-75. Afro-Arab Liver Journal 1: (1994) 137-142. Infectious Agents and Cancer 4:15: 1- 6/www.infectagentscancer.com/content/4/15. (2009) Journal of Biomedical sciences and Public Health(2015)1:3 and 9

18. Acknowledgements OM Gudza, B Chimusoro, L Mutengwa, H Mapira, N Chin’ombe, B Nherera, A Makura, K Mhandire, E Chavhunduka University of Zimbabwe, Chem Path NBSZ Harare Hospital Parirenyatwa ZACB, AFCC, IFCC Participants Publishers Sponsors