Neuregulin 4 (NRG4) - the hormone with clinical significance in gestational diabetes mellitus.

This study aimed at determining the association between serum neuregulin 4 (NRG4) and insulin in gestational diabetes mellitus (GDM) and their correlation with other metabolic parameters. A cross-sectional study was conducted from June 2018 to February 2019 at Ziauddin University, Karachi. Pregnant women (n = 80), at 24-28 weeks of gestation, were recruited. Demographic data, anthropometric measurements and antenatal history were recorded. The fasting blood was drawn for testing fasting blood sugar (FBS), NRG4, insulin and lipid profile. The subjects were evaluated for glucose challenge test after 50 g glucose ingestion. NRG4 levels (1.00 ± 0.15) were found significantly high (p < .04) in healthy group compared to GDM (0.95 ± 0.11). In GDM females, high levels for FBS (p < .02) and cholesterol (p < .03) were observed. Homeostatic model assessment of insulin resistance (HOMA-IR) had direct weak association with NRG4, inverse relationship with cholesterol and LDL but significant association (p < .05) with insulin. Significantly high NRG4 in healthy females suggests its potential role in regulating insulin sensitivity. Impact StatementWhat is already known on this subject? Neuregulin 4 (NGR4) is a batokine which plays a potential role in regulating insulin sensitivity, maintaining energy and metabolic homeostasis. Due to its role in lipid and glucose homeostasis, some studies report its role in gestational diabetes mellitus (GDM) but the results were controversial.What do the results of this study add? The study showed that NGR4 levels were significantly low in GDM subjects. Other metabolic factors especially insulin and homeostatic model assessment of insulin resistance (HOMA-IR) were found associated significantly (p < .05) with NGR4.What are the implications of these findings for clinical practice and/or further research? NGR4 can be a potential biomarker for gestational diabetes. Nevertheless, in order to confirm these findings, further studies, with bigger sample size are required to further ascertain the prospects of NRG4 as a potential biomarker for gestational diabetes.

Association of FTO variant with parental history of type 2 diabetes mellitus in adults.

OBJECTIVE: To find out the association between fat mass and obesity-associated gene polymorphism and risk factors frequently associated with type 2 diabetes mellitus. METHODS: The case-control study was conducted from January 2020 to March 2021 at the Ziauddin University, Karachi, and comprised deoxyribonucleic acid samples for fat mass and obesity-associated gene polymorphism from non-diabetic Pakistani population. Group A comprised non-diabetics with parental history of type 2 diabetes mellitus and Group B had controls without parental history of type 2 diabetes mellitus. Analysis was based on restriction fragment length polymorphism and polymerase chain reaction. Data was analysed using SPSS 25. RESULTS: Of the 150 subjects, 75(50%) each were in Group A and Group B. There were 40 (53.3%) males and 35 (46.7%) females in Group A compared to 35 (46.7%) males and 40(53.3%) females in Group B. Overall, 48% subjects were single and 52 % were married. A difference in frequency of fat mass and obesity-associated gene (rs9939609) alleles, such as TT, AA TA, was noted between the groups (p>0.999). TA allele was found to be associated with Group A (33) 44% (p=0.40), while TT allele was associated with Group B (41) 54% (p=0.414). AA allele was equally distributed between the groups (6) 8% (p=1.00). CONCLUSIONS: The TT allele of fat mass and obesity-associated gene was found to be an independent allele associated with the risk of developing type 2 diabetes mellitus.

A Systematic Review on the Mental Health Status of Patients Infected With Monkeypox Virus.

Objectives: This study aims to extract and summarize the literature on the mental health status of patients with monkeypox. Methods: This review was carried out according to the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) guidelines using different databases and publishers such as Scopus, Sage, ScienceDirect, PubMed, BMJ, Wiley Online Library, Wolters Kluwer OVID-SP, and Google Scholar. The literature review was based on monkeypox and mental health. The year of publication was 2021-2023, during the monkeypox disease period. Data were extracted from opinions, editorials, empirical studies, and surveys. Results: Based on the literature related to the mental status of patients with monkeypox, the following themes and subthemes were identified: anxiety and depression, self-harm and suicidal tendencies, neuropsychiatric symptoms, mental health, social stigma, sex workers, vaccination, and stress-related diseases. Conclusion: A review of monkeypox virus infection studies reveals that 25%-50% of patients experience anxiety and depression due to isolation, boredom, and loneliness. Factors such as infected people, a lack of competence among healthcare professionals, and shame over physical symptoms exacerbate mental insults. The implications of society include increased self-harm, suicide, low productivity, fear of stigmatization, and transmission of infection.

Identification of polymorphic alleles in TERC and TERT gene reprogramming the telomeres of newborn and legacy with parental health.

Telomere and telomerase genes (TERC and TERT) highlighted many novel genetic polymorphisms related to common diseases. This study explored the polymorphic alleles of TERC and TERT gene in parents-newborn (triad) and its association with telomere length (TL) and parental diseases (mother: Gestational Diabetes Mellitus (GDM), Preeclampsia, fathers: Diabetes, Hypertension). In this cross-sectional study, the blood samples (n = 612) were collected from parents-newborn triad (204 each) for TL (T/S ratio) quantification by using qPCR, and gene (TERC and TERT) polymorphism was detected by Sanger sequencing. The correlation analysis was used to find an association between paternal TL (T/S ratio) and newborn TL. The multivariate linear regression was applied to determine the effect of parents genes and diseases on newborn TL. A positive association (r = 0.42,0.39) (p < 0.0001) among parents and newborn TL was observed. In the diseased group, both TERC (rs10936599) and TERT (rs2736100) genes had a high frequency of allele C in newborns (OR = 0.94, P = 0.90, OR = 4.24, P = 0.012). However, among parents, TERT gene [Mother CC (B = 0.575; P = 0.196), Father CC (B = -0.739; P = 0.071)] was found significant contributing factor for Newborn TL. Diseased parents with T/T and A/C genotypes had longer newborn TL (2.82 ± 2.43, p < 0.022; 1.80 ± 1.20, p < 0.00) than the C/C genotype. Therefore, the study, confirmed that major allele C of TERC and TERT genes is associated with smaller TL in diseased parents-newborns of the targeted population.

Human papilloma virus--role in precancerous and cancerous oral lesions of tobacco chewers.

Human papilloma viruses (HPV), members of the papillomaviridae family, infects squamous epithelial cells of cevix, lower genitalia, and oral cavity. The association of HPV with oropharyngeal carcinogenesis is well documented.The incidence of oral cancer ranks second in Karachi South in both genders according to World Health Organization (WHO) statistics. This is attributed to the popularity of chewable tobacco products among the general population. Studies on Gutka-eaters in a set population of Karachi showed high frequency of HPV (17%) and high prevalence of HPV in squamous cell carcinoma in Pakistani patients (68%). The exposure of oral mucosa to chewable tobacco causes abrasions making it susceptible to HPV. This review strives to summarise the role of HPV in chewable tobacco-related precancerous and cancerous lesions. The literature of about a decade was retrieved from Google and pubMed with the under mentioned key words. It was found that the use of chewable tobacco products, especially Gutka, may increase the risk of oral squamous cell carcinoma (OSCC).

Diet Restrictions, Epigenetics and Depression.

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Near-Infrared Spectroscopy (NIRS) for Non-Invasive Diagnosis.

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Parental telomeres implications on immune senescence of newborns.

Telomere, the biological chronometer, and its effect on the immune system considerably varies among individuals. During pregnancy, multiple risk factors affect telomere reprogramming during fetal life which can lead to health disparities in newborns. These changes may cause a long-term impact on the telomere genetics of the newborn and become a reason for lifelong health implications and immune senescence. Therefore, telomere shortening in parents due to genetic variation may act as a hallmark of immune senescence and aging in their newborns.

Increasing Frequency of New Delhi Metallo-beta-Lactamase and Klebsiella pneumoniae Carbapenemase Resistant Genes in a Set of Population of Karachi.

OBJECTIVE: To determine the frequency of Klebsiella pneumoniae Carbapenemase (blaKPC) and New Delhi Metallo-Beta-Lactamase (blaNDM) resistant genes among clinical isolates of Enterobacterales in a set of Karachi population. STUDY DESIGN: An observational study. PLACE AND DURATION OF STUDY: Department of Microbiology, Dr. Ziauddin University Hospital, Karachi, Pakistan, from January 2019 to December 2020. METHODOLOGY: A total of 2100 clinical isolates of Enterobacterales were collected. All isolates of Carbapenem-Resistant Enterobacterales (CRE) (Escherichia coli, Enterobacter and Klebsiella species) on the basis of Meropenem screening test positivity were included in the study. DNA was extracted and PCR was performed for resistant genes detection. Frequencies and percentages were computed for categorical variables and mean values and standard deviation for quantitative variables. RESULTS: Among 2100 isolates of Enterobacterales, the majority were E. coli 1260 (60%), followed by Klebsiella species 462 (22%), and Enterobacter species 210 (10%). The sources of CRE isolates included 34 (25%) from respiratory (tracheal aspirate, pleural fluid, and gastric lavage); 33 (24.26%) urine, 32 (25.53%) pus, 15 (11.03%) blood, and 20 (14.7%) others (ascitic fluid, stents, and tissue). All isolates of CRE were sensitive (100%) to Colistin, Tigecycline and Fosfomycin. Biochemically confirmed CRE 136 (6.5%) isolates, (79 (58%) males and 57 (42%) females), were selected for detecting resistant genes. The PCR showed 32 (23.52%) positive for both NDM and KPC resistant genes, 28 (20.58%) for NDM and 19 (13.97%) for KPC alone. Out of 79 followed up patients, 58 (73.4%) expired while 21 (26.6%) were discharged. CONCLUSION: The frequency of blaNDM and blaKPC resistant genes in CRE isolates depicted increasing trend. Colistin, Fosfomycin, and Tigecycline showed high antimicrobial sensitivities in vitro. Further measures need to be applied for CRE with comprehensive resistant genes detection to curtail antimicrobial resistance. KEY WORDS:  Frequency, KPC, NDM, Klebsiella species, Carbapenemases, Enterobacterales E.coli.

Comparison of KRAS gene in circulating tumor DNA levels vs histological grading of colorectal cancer patients through liquid biopsy.

Background: To determine KRAS gene in circulating tumor DNA in comparison with histological grading through liquid biopsy in colorectal cancer patients. Methods: This dual-centered cross-sectional study included 73 diagnosed patients of colorectal cancer at different grading levels [Grade I, well differentiated (n = 7, 9.5%); Grade II, moderately differentiated (n = 14,18.9%); and Grade III, poorly differentiated (n = 52, 70%)]. Blood was collected, and plasma was separated. ctDNA was extracted, using magnetic bead-based technique (MagMAX Cell-Free DNA kit). KRAS gene was quantified through qPCR. STRING database was used to find KRAS interactomes. Results: Mean threshold cycle (CT value) of KRAS gene in Grade III samples showed significantly higher (P = 0.001) levels of ctDNA (2.7 ± 1.14) compared with Grade II and Grade I (3.1 ± 0.68, 2.3 ± 0.60), respectively. Grading characterization showed that rectal cancer (n = 22, 42.3%) with Grade III (68.8%) was more prevalent than colon and sigmoid cancer (n = 19, 36.5%, n = 11, 21%, respectively). STRING database showed 10 functional genes interacting with KRAS expressed as gene/proteins. Conclusion: Liquid biopsy can be used to detect ctDNA in plasma of CRC patients and enabled to detect the KRAS gene by qPCR. The technique being less invasive and cost-effective is convenient for multiple biopsies in different cancers.

Fok1 VDR Gene Polymorphisms as the Risk factor for Diabetes Mellitus.

OBJECTIVE: To investigate the prevalence of single nucleotide polymorphisms of vitamin D receptor (VDR) gene, and its association with Type 2 Diabetes Mellitus. STUDY DESIGN:  Cross-sectional study. PLACE AND DURATION OF STUDY: Ziauddin University Hospital, Karachi, from January 2018 to 2020. METHODOLOGY: A total of 200 unrelated individuals, aged 25 to 65 years, were selected and divided into two groups, T2DM patients (100) and non-diabetic controls (100). After consent, demographics, diabetic history and related risk factors were recorded in a standard questionnaire and blood was collected. The VDR (Fok1, Taq1 and Apa1) polymorphisms were analyzed through PCR and RFLP. RESULTS: In T2DM patients, F/F and F/f genotypes were found in 49 (49%) and 45 (45%) patients, respectively. The ff genotype was less common in T2DM [6 (6%)], compared to healthy controls [14 (14%)], (odds ratio=0.473, 95% CI: 0.267-0.839). The frequency of T/t genotype was 36% and 31% in the control and diabetic group respectively while for A/a genotype was 38% and 34% respectively, the results were not statistically significant. CONCLUSION: T2DM was found significantly associated with Fok1 Polymorphisms of VDR gene. The study found a protective role of ff of Fok1 in diabetic patients. Further studies in larger cohorts are required for validation. KEY WORDS: Polymorphism, Restriction fragment length, Receptor, Vitamin D3 receptor, Type 2 diabetes mellitus.

Parental Genetics Communicate with Intrauterine Environment to Reprogram Newborn Telomeres and Immunity.

Telomeres, markers for cellular senescence, have been found substantially influenced by parental inheritance. It is well known that genomic stability is preserved by the DNA repair mechanism through telomerase. This study aimed to determine the association between parents−newborn telomere length (TL) and telomerase gene (TERT), highlighting DNA repair combined with TL/TERT polymorphism and immunosenescence of the triad. The mother−father−newborn triad blood samples (n = 312) were collected from Ziauddin Hospitals, Pakistan, between September 2021 and June 2022. The telomere length (T/S ratio) was quantified by qPCR, polymorphism was identified by Sanger sequencing, and immunosenescence by flow cytometry. The linear regression was applied to TL and gene association. The newborns had longest TL (2.51 ± 2.87) and strong positive association (R = 0.25, p ≤ 0.0001) (transgenerational health effects) with mothers' TL (1.6 ± 2.00). Maternal demographics­socioeconomic status, education, and occupation­showed significant effects on TL of newborns (p < 0.015, 0.034, 0.04, respectively). The TERT risk genotype CC (rs2736100) was predominant in the triad (0.6, 0.5, 0.65, respectively) with a strong positive association with newborn TL (ß = 2.91, <0.0011). Further analysis highlighted the expression of KLRG 1+ in T-cells with shorter TL but less frequent among newborns. The study concludes that TERT, parental TL, antenatal maternal health, and immunity have a significantly positive effect on the repair of newborn TL.

The calcium connection--deficient mothers with normal newborn.

A cross-sectional study on blood calcium levels of pregnant women of low socioeconomic status was designed to find out the consequences of low levels on the outcome of pregnancy. Blood samples from 52 women and their cord were collected at the time of delivery. Calcium was analyzed calorimetrically in maternal as well as cord blood. For comparative analysis, levels of phosphorus were analyzed on atomic absorption spectrophotometer. The results were compared with 24 nonpregnant age matched controls. Calcium levels were significantly low (p<0.01), in maternal (8.13±0.40 mg/dl) and cord blood (8.69±0.28 mg/dl) compared to controls (10.5±0.6 mg/dl) and significantly higher in primiparous mothers (M=8.2±0.78 mg/dl,C=8.1±0.50 mg/dl) compared to multiparous (M=6.4±0.59 mg/dl,C=7.9±0.53 mg/dl). Calcium levels were also found significantly lower in mothers who had a miscarriage (M=7.60±0.37 mg/dl,C=9.1±0.65 mg/dl) before this pregnancy than the primiparous and the controls. The results on the whole suggest that women tend to develop a marginal calcium deficiency after pregnancy.

Detection of Kras Gene in Colorectal Cancer Patients through Liquid Biopsy: A Cost-effective Method.

OBJECTIVE: To detect the Kras gene through liquid biopsy, a less invasive technique in diagnosed colorectal cancer patients. STUDY DESIGN: Cross-sectional study. PLACE AND DURATION OF STUDY: Department of Oncology, Dr. Ziauddin Hospital and Bait-us-Sukoon Cancer Hospital, Karachi, from 2019 to 2020. METHODOLOGY: Circulating tumor DNA (ctDNA) in colorectal cancer patients was extracted through magnetic bead technique using MagMAX cell free DNA kit (Thermofisher, Uk). The frequency of Kras gene was quantified using a real-time polymerase chain reaction (RT-PCR) assay (qPCR). ANOVA and Chi-square tests were utilised for statistical analysis. RESULTS: Mean threshold cycle (CT) of Kras gene showed significantly higher expression 15.6 ± 1.82 (p=0.001) in stage IV CRC cases compared to early stages (19.53 ± 18.223.7 ± 2.9 and 19.8 ± 2.69 of stage 1, 2 and 3, respectively. Similarly, ΔCT mean of Kras gene at stage IV showed significantly higher expression of 2.48 ± 1.40 (0.048), compared to 2.39 ± 0.6, 3.12 ± 0.68 and 3.15 ± 0.41 of stage 1, 2 and 3, respectively. Males (n=40, 55%) showed significant association (p=0.001) with CRC compared to females (n=33, 45%). Categorisation of tumor types within different age groups revealed that colon cancer was more frequent (n=11, 15.1%) in the 41-50 age group, while rectal cancer was more frequent (n= 11, 15.1%) in the 41-50 age group, while rectal cancer was more in the 51-60 age group (n=11, 15.1%). CONCLUSION: Kras gene was detected with significantly increased levels in plasma of CRC patients at advanced stages. This confirms that liquid biopsy can be used to detect Kras gene in ctDNA of CRC patients through a magnetic bead based technique. Key Words: Liquid biopsy, Circulating tumor DNA, KRAS, Colorectal cancer, Real-time polymerase chain reaction.

Expression of DnMTs and MBDs in AlCl3-Induced Neurotoxicity Mouse Model.

Alteration in DNA methylation after aluminum exposure has been shown to contribute in pathogenesis of Alzheimer's disease (AD). This study is aimed to determine the effect of Al exposure (42 and 60 days) on learning and memory and the expression of proteins involved in DNA methylation (MBD1, MBD2, MBD3, MeCP2 (methyl CpG binding protein 2), DnMT1 and DnMT3a). Male BALB/c mice were treated with AlCl3 for either 42 days or 60 days. After treatment completion, learning and memory were compared to the control group using novel object recognition test, elevated plus maze test, open field test, and Morris water maze test. The treated animals and their respective controls were sacrificed after cognitive testing and samples from their whole cortex and hippocampus were harvested for gene expression analysis. Mice treated with AlCl3 showed significant cognitive deficit with impaired short-term memory, elevated anxiety, and deterioration in spatial and reference memory. The AlCl3 treatment showed significant reduction in the expression of MBDs in the whole cortex at 60 days of treatment as compared to control. AlCl3-treated animals showed decreased expression of MBDs and DnMT3a in the hippocampus for longer treated animals but strikingly, MBD2 showed significantly increased expression in AlCl3-treated animals at 60 days p ≤ 0.001. In conclusion, this study showed that AlCl3-treated animals showed significant memory and cognitive deficits and it is associated with significant changes in the expression of proteins involved in DNA methylation mechanism. Moreover, different Al exposure duration had slightly different effects.

Association of the JAZF1 Variant in Adults With a Parental History of Type 2 Diabetes Mellitus In Pakistan.

Background Type 2 diabetes mellitus (T2DM) is a chronic multifactorial condition and quickly growing disease in Pakistan. Many genes together with Zinc finger protein 1 (JAZF1) have already been described earlier in the literature but the role of JAZF1 in this subset of the population is yet to define. This study was aimed at identifying JAZF1 polymorphism and the risk of developing T2DM in persons with a parental history of T2DM in the Pakistani population. Methods DNA samples from 75 non-diabetic Pakistani participants with a family history of T2DM and 75 controls were evaluated by using a polymerase chain reaction (PCR) and the restriction fragment length polymorphism method. Results The alleles AA and AG and the GG genotype of JAZF1 (rs864745) varied considerably in frequency distribution between cases and control (p<0.05). The GG was independently and significantly associated with cases who had a family history of T2DM [odds ratio (OR) 2.6 (95% confidence interval (Cl) 1.3-5.1); p=0.005] while the AA allele was significantly associated with controls without a family history of T2DM [odds ratio (OR) 0.39 (95% confidence interval (Cl) 0.2-0.7); p=0.0059] and the allele AG has no significance and was equally distributed among control and cases with p-value=1.000. Conclusion Genotype GG of the JAZF1 variant was found significantly associated with the risk of developing type 2 diabetes mellitus in the Pakistani subset of the population.

Association of Zinc Transporter-8 Autoantibody (ZnT8A) with Type 1 Diabetes Mellitus.

BACKGROUND: Zinc transporter 8 autoantibody (ZnT8A), discovered through bioinformatics, is identified as another major biomarker for type 1 diabetes mellitus (T1DM), expanding the panel of diagnostic autoantibodies. The absence of standard autoantibodies in T1DM patients and the presence of ZnT8A in individuals before disease development has led the researchers to evaluate ZnT8A to gather information about the frequency and its association. Therefore, we aim to find out the concentration of ZnT8A and its association with T1DM. METHODS: A case-control study with 25 type 1 diabetes mellitus patients and 25 first-degree relatives of cases as controls was conducted at Ziauddin University in collaboration with the Baqai Institute of Diabetology and Endocrinology (BIDE), Karachi. Demographic data were collected from patients on a standard questionnaire. Blood samples were collected, after approval from Ziauddin Ethics Review Committee, from subjects and serum was separated to estimate ZnT8A by using sandwich enzyme-linked immunosorbent assay (ELISA). RESULTS: The mean age at diagnosis of T1DM patients was 13.40±5.05 years, and the duration of diabetes was 7.74±5.85 years. The frequency of ZnT8A was found higher in cases (19 (76%)) compared to controls (6 (24%)). ZnT8A concentrations were significantly higher in cases (13.82 ng/ml) compared to the controls (8.78 ng/ml; p= 0.024). The cut-off value of 9 ng/ml was selected for measuring sensitivity, specificity, and accuracy, which were determined as 76%, 76%, and 76%, respectively. CONCLUSIONS: ZnT8A was found significantly associated with T1DM. Subjects with ZnT8A values ≥ 9 ng/ml are 10 times more at risk to develop T1DM (p = 0.000).

Gender disparity in infections of Hepatitis B virus.

Gender differences prevail in the infections caused by the Hepatitis B virus. Four hundred and seventy two patients with HBV infection were selected for the study. The frequency of hepatic infection in males was 79.5% (n=375) and in females 20.5% (n=97), with a male to female ratio of 3.8:1. Out of 472 patients, 49% had acute hepatitis, 26% were carriers, 18% had chronic hepatitis, 6% had cirrhosis and 3% patients had hepatocellular carcinoma. Male dominance was found to be consistent in all categories of patients. When the patients were divided into groups according to age, the male to female ratio increased during the reproductive years. There may be an influence of estrogen in the protection and defense of hepatic cells against the development of chronic liver disease.

Cell death induced by Morarah and Khaltita in hepatoma cancer cells (Huh-7).

OBJECTIVE: To compare the combined and isolated growth inhibitory effects of Morarah and Khaltita (herbs) on hepatoma cell lines (Huh-7), through induction of apoptosis or necrosis. STUDY DESIGN: Comparative controlled in-vitro study. PLACE AND DURATION OF STUDY: The Molecular Biology Laboratory, The Aga Khan University, Karachi, from June to December 2006. METHODOLOGY: The growth of hepatoma cell lines (Huh-7) was checked by adding Khaltita and Morarah to the cells before culture in a 24 well plate. Six wells were selected and labeled for each of the four variables (controls, Khaltita, Morarah and mixture). After 2 days, cells were studied under an inverted phase contrast microscope and fields were recorded. Approximately four fields per slide of higher intensity were selected randomly to determine the dead cell density, and the procedure was repeated 10 or more times. Frequency and percentages were calculated for dead or alive cells in controls, Morarah, Khaltita and their mixture. Chi-square was used to compare the qualitative variables. P-values < 0.05 were considered significant. RESULTS: Morarah and Khaltita were found to induce statistically significant (p < 0.001) cell death in hepatoma cell lines (Huh-7). At a magnification of 40x, the controls showed 1% dead cells compared to 91% in Morarah, 83% in Khaltita and 73% in combined mixture of Khaltita and Morarah. At magnification of 20x, the controls showed 4% dead cells compared to 44% in Morarah, 47% in Khaltita and 49% in the combined mixture of Khaltita and Morarah. CONCLUSION: Morarah and Khaltita induced cell death in cultured hepatoma cells (Huh-7).

Frequency of hepatitis C and D super infection in patients with hepatitis B related complex liver disorders.

OBJECTIVE: To determine the frequency of super infection of hepatitis C and D in patients with hepatitis B related complex liver disorders and the distribution of HBV genotypes in these patients. STUDY DESIGN: Descriptive study. PLACE AND DURATION OF STUDY: The Gastroenterology Unit of PMRC in JPMC, Karachi, from July 2006 to June 2007. METHODOLOGY: All patients registered for HBV associated infections were selected. Blood was drawn from 180 patients who fulfilled the inclusion criteria. Those with an incomplete test profile were excluded. All clinical conditions were investigated through liver function tests, coagulation profile, and findings at abdominal ultrasonography, upper gastrointestinal endoscopy and liver biopsy. Liver cirrhosis and hepatocellular carcinoma (HCC) were diagnosed either on the basis of histology, or on a combination of radiological, endoscopic and laboratory data. Hepatitis B virus DNA was extracted from serum, and subjected to a nested PCR using the type specific primers for HBV genotype. Descriptive statistics were used for frequency and mean determination. RESULTS: The 129 patients finally selected for statistical analysis included 108 (84%) males and 21 (16%) females. The age ranged from 6- 68 years (mean=31.5 +/-12.39 years). There were 70 (54.2%) patients of non-cirrhotic, chronic hepatitis (CLD), 38 (29.4%) carriers, 12 (9.3%) cirrhotics and 9 (6.9%) HCC patients. Among the 129 patients, 45 (34.9%) were positive for double infection with HDV. These included 35 CLD cases, 7 cirrhotic and 3 carriers, 4 (3.1%) patients were positive for double infection with HCV including one with CLD, 2 with cirrhosis and one with HCC. Triple infection with HBV/HDV/HCV was present in 4 (3.1%) patients who had CLD. Approximately 59% (n=76) patients were not coinfected, though 9 had developed HCC. The genotype distribution of HBV was observed as D in 98 (76%) patients, A in 24 (18.6%), and AD mix in 7 (5.4%). Genotypes B, C, E or F were not found. Accordingly, genotype D strains were the predominant strains among all categories. CONCLUSION: The frequency of super infection of hepatitis C and D was found to be highest in HBV cirrhosis patients compared to patients having chronic liver disease (non-cirrhotics) and carriers. Genotype D of hepatitis B virus was found dominant in all hepatitis B related complex liver disorders.