Clinical Significance of Anti-Ribosomal P Protein Antibodies in Patients with Lupus Nephritis.

BACKGROUND: Systemic Lupus Erythematosus (SLE) is a chronic inflammatory disorder affecting multiple systems of the body. Clinical features show wide variations in patients with the different ethnic background. Renal involvement is a predictor of poor prognosis. Immunological workup is an integral part of SLE diagnostic criteria. Anti-ribosomal P Protein (anti-P) antibodies are highly specific for SLE. They may be present in Antinuclear Antibodies (ANA) negative SLE patients. Their role in Lupus Nephritis (LN) is under debate, some researchers found them associated with poor prognosis whereas others found favorable effect of these antibodies on renal disease. OBJECTIVE: In this study, we investigated frequency of anti-P antibodies and the effect of these antibodies on renal functions in the LN patients. METHODS: A total of 133 SLE patients were enrolled in this study. All patients had ANA in their sera. Anti-P antibodies along with other autoantibodies against extractable nuclear antigens (anti-Sm, anti- SS-A, anti-SS-B, anti-histones and anti-RNP) were detected by Immunoblot assay. Anti-dsDNA antibodies were detected by indirect Immunofluorescence Assay (IFA). RESULTS: We found anti-P antibodies in 10.5% LN patients. Interestingly their presence in association with anti-dsDNA was associated with improved renal functions in comparison to those who had antidsDNA antibodies in isolation (serum creatinine: 1.3 ± 0.8 mg/dl vs. 3.0 ± 3.0; P= 0.091). CONCLUSION: Anti-dsDNA antibodies are directly involved in renal pathology in SLE patients. As these antibodies are nephrotoxic, concomitant occurrence of anti-P antibodies seems to offer a shielding effect on renal functions, which was evident by normal serum creatinine levels. Therefore, anti-P antibodies may be considered as a good prognostic marker in these patients.

Plasma cell-rich acute rejections in living-related kidney transplantation: a clinicopathological study of 50 cases.

BACKGROUND: Acute rejections (ARs) with plasma cell-rich infiltrates (PCARs) are associated with poor outcomes. PATIENTS AND METHODS: Between February 2012 and December 2013, 1630 dysfunctional renal graft biopsies were performed. Of these, 50 (3%) showed PCAR. ARs with >10% plasma cells were defined as PCAR. Human leukocyte antigen (HLA) antibodies were tested in historic sera and at the time of PCAR. Treatment for PCAR comprised methylprednisolone, antithymocyte globulin, plasmapheresis, and anti-CD20 antibody. RESULTS: Of the 1630 dysfunctional biopsies, 50 (3%) had PCAR which occurred 3.1 ± 2.55 yr after transplant. The percentage of plasma cells was 28.8 ± 11.7, and CD138, 29.0 ± 12.4. Donor-specific antibodies (DSAs) were found in 32 (64%) overall, Class I in 15% and Class II in 65%. Post-treatment serum creatinine improved from 3.80 ± 2.59 to 2.66 ± 1.59 mg/dL in DSA positive (p < 0.003) and from 2.59 ± 1.09 to 2.08 ± 0.86 mg/dL in DSA negative (p < 0.008). One- and two-yr graft survival after PCAR was 72%, 42% in the DSA-positive vs. 89%, 82% in the DSA-negative group, respectively (p = 0.071). CONCLUSIONS: Our results show that PCAR occurs late after transplant and in many cases is associated with DSAs. Graft outcome was poor when PCAR was associated with DSAs.

Vasculitis with renal involvement in essential mixed cryoglobulinemia: Case report and mini-review.

The discovery of a strong association between hepatitis C virus (HCV) infection and mixed cryoglobulinemia (MC) has led to an increasingly rare diagnosis of idiopathic essential MC (EMC). The incidence of EMC is high in regions where there is a comparatively low HCV infection burden and low in areas of high infection prevalence, including HCV. The diagnosis of EMC requires an extensive laboratory investigation to exclude all possible causes of cryoglobulin formation. In addition, although cryoglobulin testing is simple, improper testing conditions will result in false negative results. Here, we present a 46-year-old female patient with a case of EMC with dermatological and renal manifestations, highlighting the importance of extensive investigation to reach a proper diagnosis. We review the need for appropriate laboratory testing, which is often neglected in clinical practice and which can result in false negative results. This review also emphasizes the significance of an extended testing repertoire necessary for better patient management. Despite a strong association of MC with HCV infection and other causes that lead to cryoglobulin formation, EMC remains a separate entity. Correct diagnosis requires proper temperature regulation during sample handling, as well as characterization and quantification of the cryoprecipitate. Inclusion of rheumatoid factor activity and complement levels in the cryoglobulin test-panel promotes better patient management and monitoring. Consensus guidelines should be developed and implemented for cryoglobulin detection and the diagnosis of cryoglobulinemic syndrome, which will reduce variability in inter-laboratory reporting.

Clinical Profile and HLA Typing of Autoimmune Hepatitis From Pakistan.

BACKGROUND: Human leukocyte antigen (HLA) typing in autoimmune hepatitis (AIH) has been investigated in different populations and ethnic groups, but no such data is available from Pakistan. OBJECTIVES: The aim of this study was to evaluate the clinical profile of autoimmune hepatitis (AIH), and determine the associated antigens and alleles by performing HLA typing. PATIENTS AND METHODS: A total of 58 patients, diagnosed and treated as AIH in the last 10 years were reviewed. Diagnosis was based on International AIH Group criteria. Forty one patients underwent liver biopsy. HLA typing was performed in 44 patients and 912 controls by serological method for HLA A and B, and by PCR technique using sequence specific primers for DR alleles. RESULTS: Of 58 cases, 35 were females (60.3%). The median age was 14.5 (range 4-70 years), and AIH score was 14 (10-22). Thirty-six (62.0%) patients had type 1 AIH, 10 (17.2%) type 2, and the remaining 12 were seronegative with biopsy proven AIH. Forty-nine patients (84.4%) had cirrhosis. Twenty-four (41.4%) patients had ascites at the time of presentation. Among 41 patients who underwent liver biopsy, thirty-two had advance stages III and IV disease, and twenty had severe grade of inflammation. Fifteen patients had other associated autoimmune diseases and one developed hepatocellular carcinoma. HLA A2 (P = 0.036), HLA A9 (23) (P = 0.018), HLA A10 (25) (P = 0.000), HLA A19 (33) (P = 0.000), HLA B15 (63) (P = 0.007), HLA B40 (61) ( P = 0.002), HLA DR6 (P = 0.001) with its subtypes HLA-DRB1*13 (P = 0.032) and HLA-DRB1*14 (p = 0.017) were more prevalent in AIH with statistical significance than controls. CONCLUSIONS: AIH in our region presents with advanced disease affecting predominantly children and adolescents. There is a genetic association of HLA DR6 along with other alleles and antigens in our patients with AIH.

Prevalence of anti- beta2GPI antibodies and their isotypes in patients with renal diseases and clinical suspicion of antiphospholipid syndrome.

BACKGROUND: Antiphospholipid antibodies (aPL) are autoantibodies that are associated with a clinical state of hypercoagulability and diverse clinical manifestations collectively known as antiphospholipid syndrome (APS). OBJECTIVES: To investigate the prevalence of anti-beta2glycoproteinI-antibodies (anti-ß2GPI) and their isotypes in patients with renal diseases and clinical suspicion of antiphospholipid syndrome (APS). PATIENTS AND METHODS: This is a retrospective study in which we have analyzed the prevalence of anti-ß2GPI and its isotypes in 170 patients on initial testing and in 29 patients repeated after 12 weeks for confirmation of APS.  The clinical information was provided by the treating physicians or retrieved from the clinical records. The tests for anti-ß2GPI screening and its isotypes (IgG, IgM and IgA) detection were assessed. RESULTS: On initial samples, anti-ß2GPI was positive in 118patients.  IgA-ß2GPI positivity (93; 79%) was significantly higher than IgM and IgG isotypes.  Out of anti-ß2GPI positive patients, clinical features in 95 patients were suggestive of APS or had SLE.  Of these, IgA isotypes was found in 66% (P = 0.010), IgM in 31% (P = 0.033), and IgG in 11% (P = 0.033). On repeat testing, anti-ß2GPI was persistently found In 22 patients with a continual predominance of IgA-anti-ß2GPI over IgM and IgG isotypes (91% vs. 45.5% and 18% respectively). CONCLUSIONS:   Our results show that IgA-anti-ß2GPI antibodies are the most prevalent isotypes in patients with renal disease or on renal replacement therapy in our population.  Thus inclusion of IgA-anti-ß2GPI in the testing repertoire may increase the diagnostic sensitivity for APS in patients with renal diseases.

Coeliac disease and multiple immunodeficiencies: case report of a diagnostic dilemma.

Coeliac disease (CD) often coexists with other autoimmune and primary immunodeficiency diseases (PID), creating a problem in timely diagnosis and management. An unusual case of coeliac disease that was difficult to diagnose and manage because of its unusual clinical presentation. Initially diagnosed as celiac disease but showed poor response to standard therapy is reported. Frequent attacks of opportunistic infections led to immunodeficiency work-up that revealed natural killer cell (NK) deficiency with low serum IgA and IgG2 levels. The patient eventually succumbed to recurrent infections. The co-existence of PID is unusual in a patient with CD. This case report highlights the importance of investigating PID in patients with autoimmunity.

Curriculum guide for research ethics workshops for countries in the Middle East.

To help ensure the ethical conduct of research, many have recommended educational efforts in research ethics to investigators and members of research ethics committees (RECs). One type of education activity involves multi-day workshops in research ethics. To be effective, such workshops should contain the appropriate content and teaching techniques geared towards the learning styles of the targeted audiences. To ensure consistency in content and quality, we describe the development of a curriculum guide, core competencies and associated learning objectives and activities to help educators organize research ethics workshops in their respective institutions. The curriculum guide is divided into modular units to enable planners to develop workshops of different lengths and choose content materials that match the needs, abilities, and prior experiences of the target audiences. The content material in the curriculum guide is relevant for audiences in the Middle East, because individuals from the Middle East who participated in a Certificate Program in research ethics selected and developed the training materials (e.g., articles, PowerPoint slides, case studies, protocols). Also, many of the activities incorporate active-learning methods, consisting of group work activities analyzing case studies and reviewing protocols. The development of such a workshop training curriculum guide represents a sustainable educational resource to enhance research ethics capacity in the Middle East.

Pattern of morphology in renal biopsies of nephrotic syndrome patients. Correlation with immunoglobulin and complement deposition and serology.

OBJECTIVE: To determine the morphological pattern in renal biopsies from nephrotic patients and their correlation with immunoglobulin and complement deposition as detected by immunofluorescence microscopy (IMF) and serology to arrive at correct diagnosis of glomerulonephritis (GN). METHODS: The study was carried out at the departments of Immunology and Histopathology, Sindh Institute of Urology and Transplantation (SIUT) from April 2007 to March 2008. A total of 200 patients, including children and adults were included. All patients presented with nephrotic syndrome (NS). Clinical and laboratory investigations including serology were noted, renal biopsies performed and studied by light and immunofluorescence microscopy (IMF). RESULTS: Of 200 patients, 74 (37%) were children (< or = 18 years) and 126 (63%) adults (> or = 19 years). Mean age of children was 11.34 +/- 4.85 years (range 3-18 years) and that of adults was 35.44 +/- 11.4 years (range: 19-70 years). The total percent change in L/M diagnosis after serological tests was 11.5% and after IMF studies, 23.5%. Combined serologic and IMF studies lead to 35% change in L/M diagnosis of the renal biopsies in nephrotic syndrome patients. CONCLUSION: Our results demonstrate that the ancillary techniques of IMF microscopy and serology are indispensable in the elucidation of final specific diagnosis causing nephrotic syndrome in a substantial number of cases and these should be employed routinely in the pathologic evaluation of renal biopsies. The study emphasizes the importance of combined approach in the investigation of renal biopsies in nephrotic syndrome.

Molecular characterization of Pakistani field isolates of infectious bursal disease virus.

The reverse transcriptase-polymerase chain reaction followed by restriction fragment length polymorphism (RT-PCR/RFLP) technique was used to identify and characterize Pakistani field isolates of infectious bursal disease virus (IBDV). These isolates have caused heavy losses to the poultry industry (mortality up to 60%) during the period between 1999 and 2005. Ten samples (five local isolates and five commercial vaccines) were examined for IBDV. Nine samples were positive for IBDV as evidenced by the amplification of the 743-bp region of the VP2 gene by RT-PCR. The RT-PCR products were subjected to restriction enzyme digestion with BstNI, MboI, and SspI. The RFLP profiles of all samples on digestion with the MboI enzyme yielded a fragment size of 229 and 362 bp except for vaccine strain Bursine Plus, which yielded a profile of 229 and 480 bp. However, digestion with BstNI yielded two distinct RFLP patterns. The first profile was detected in field isolates ML-1/SPVC/2001 and NP2/SPVC/2002 with four fragments of 119, 154, 172, and 209 bp, resembling RFLP profiles of molecular group 4 isolates. NL-3/SPVC/2003, NK-4/SPVC/2004, and NPK-5/SPVC/2005 generated a different RFLP profile with fragments of 119, 172, and 424 bp, resembling the profiles of molecular group 6 isolates. However, all the field and vaccine strains showed the absence of SspI restriction sites in their genome. It can be concluded that the Pakistani isolates can be grouped in molecular groups 4 and 6 of IBDV.

Evaluation of pre transplant T-cell activation status by soluble CD 30 determination.

OBJECTIVE: To evaluate the utility of serum CD30 (sCD30) levels as predictor of early acute graft rejection in live related renal transplant programme. METHODS: This prospective study included 50 consecutive renal transplant recipients who received their first live related renal allograft at the Sindh Institute of Urology and Transplantation (SIUT) between October 2006 and March 2007. Blood samples were obtained one day before transplantation and on the third and fourteenth posttransplant days. Blood samples were also obtained from 50, age and sex matched healthy control individuals. Levels of serum sCD30 were measured by Enzyme Linked Immunosorbent Assay (ELISA). RESULT: Donor-recipient blood group matching was identical in all patients. Pre-transplant lymphocyte crossmatch for T and B cells was negative, and panel reactive antibodies (PRA) were 0% for all recipients. The mean age of recipients was 31.6 +/- 10.23 years (range 5 to 55 years), while mean donor age was 32.74 +/- 8.48 years (range 21-50 years). Eleven (22%) recipients and donors were HLA identical while remaining (78%) were one haplotype match. Average serum sCD30 pre-transplant levels (37.8 +/- 4.97U/ml) were significantly higher than those of healthy individual's mean value of 8.48 +/- 4.97 U/ml, (P = 0.001). Eight (16%) patients developed acute rejection episode during this follow up period. Rejections were described and classified according to BANFF 97 classification. CONCLUSION: In this small single center study the serum levels of sCD30 did not show any significant difference between rejection and non rejection group in our transplant population.

National study of reproductive tract infections among high risk groups of Lahore and Karachi.

OBJECTIVE: To estimate the prevalence of reproductive tract infections (RTIs) among the Female Sex Workers (FSWs), Male Sex Workers (MSWs), Injecting Drug Users (IDUs) and truck drivers. STUDY DESIGN: Cross-sectional. PLACE AND DURATION OF STUDY: Lahore and Karachi, from March to August, 2004. METHODOLOGY: Four hundred FSWs, 400 MSWs, 200 eunuchs, 400 IDUs and 400 truck drivers were interviewed and examined. Biological testing included PCR and ELISA. RESULTS: The mean prevalence of syphilis was 17.7% (95% CI: 15.1-20.3%). Genital gonorrhoea was found among 0.8-12.3% of subjects with mean of 4.5% (95% CI: 0.8-5.2%). Genital chlamydia was seen in 0.2-11% individuals with mean of 2.6% (95% CI: 2.1-3.1%). Trichomonas was present in 19.3% FSWs of Lahore and 5.52% FSWs of Karachi. Only 0.8% truckers in Lahore had trichomonas. In Lahore, 47.6% FSWs and in Karachi, 27.4% FSWs were positive for bacterial vaginosis. A vast majority of IDUs, 91.8% in Lahore and 87% in Karachi were hepatitis C positive. CONCLUSION: The prevalence of syphilis is very high among all high-risk groups; particularly so among eunuchs (60.2% in Karachi and 32.3% in Lahore). Such a high levels of RTIs indicate a serious threat for HIV epidemic because of socially transmitted infection.

High prevalence of HIV infection among injection drug users (IDUs) in Hyderabad and Sukkur, Pakistan.

OBJECTIVE: To estimate size of injection drug users (IDU), determine their high risk behaviours and assess the prevalence of HIV. METHOD: As part of second generation surveillance (SGS), we investigated specific demographic and behavioural characteristics of IDUs in Hyderabad and Sukkur in 2005. It was a cross sectional study. The survey was preceded by geographic mapping to determine size estimation and to define sampling procedures prior to integrated behavioural and biological survey (IBBS). A sample size of at least 400 was calculated for each city. Besides calculating frequencies, chi square was used for comparing variables among HIV positive and negative IDUs like time elapsed as IDU, number of injections, sharing needles and self perception of acquiring HIV infection. RESULTS: A total of 800 (Hyderabad 398; Sukkur 402) questionnaires and DBS samples were collected. The estimated number of IDUs in both cities was 3,225 (Hyderabad 975 and Sukkur 2250 respectively). Average age of IDUs in Hyderabad was 36.5 years and 34.6 years in Sukkur. Sharing of injection equipment for last injection was reported by 34 (8.5%) in Hyderabad and 135 (33.6%) in Sukkur. In both cities behaviours such as injecting drugs for more than 10 years (p = 0.00) and injecting four or more times in a day (p = 0.11) were significantly associated with seropositivity of HIV infection. In Hyderabad the seroprevalence of HIV was 25.4% (101/398) and in Sukkur it was 19.2% (77/402). CONCLUSION: The burden of HIV among IDUs in Hyderabad and Sukkur is extremely high and can play a significant role in transmitting the infection to other vulnerable groups.

Commercial transplants in local Pakistanis from vended kidneys: a socio-economic and outcome study.

Donor shortage and absence of transplant law lead to unrelated commercial transplants in Pakistan. We report the socio-economic and outcome parameters of 126 local recipients of unrelated kidney vendor transplants presenting to our institute between 1997 and 2007. Their outcome was compared with 180 recipients of living-related donor transplants matched for age, gender and transplant duration as controls. Age of commercial recipients was 35.63 +/- 11.57 years with an M:F ratio of 2.4:1. Majority (92%) were transplanted in northern Pakistan paying US$7271 +/- 2198. All were educated with 50% being graduates or above and rich earning a monthly salary of US$517 +/- 518 with 44% earning >US$500. Comparison of commercial recipients with controls showed high comorbidities 35 (28%) vs. 14 (8%) (P = 0.0001) with diabetes, hepatitis-C and cardiovascular diseases. Donor age was 29.97 +/- 6.16 vs. 32.63 +/- 9.3 years (P = 0.035). Biologic agents induction in 101 (80%) vs. 14 (8%) (P = 0.0001), acute rejections in 42 (33%) vs. 31 (17%) (P = 0.005), 1-year creatinine 1.84 +/- 1.28 vs. 1.27 +/- 0.4 mg/dl (P = 0.0001), surgical complications 28 (22%) vs. 14 (8%) (P = 0.001), tuberculosis 14 (11%) vs. 6 (6%) (P = 0.007), acute hepatitis 20 (16%) vs. 3 (2%) (P = 0.0001), cytomegalovirus 33 (26%) vs. 21 (11%) (P = 0.001) and recurrent urinary tract infection 35 (28%) vs. 30 (16%) (P = 0.034). Overall 1- and 5-year graft survival was 86% and 45% vs. 94% and 80%, respectively (P = 0.00001). Total deaths were 34 (27%) vs. 12 (6.0%) (P = 0.001). In conclusion, recipients of the vended kidneys are poor candidates, educated, rich and often self-selecting. Their outcome is poor, which will leave them poorer still and back to dialysis if not death.

Celiac disease in children with persistent diarrhea and failure to thrive.

OBJECTIVE: To determine the frequency of celiac disease (CD) in children with failure to thrive (FTT) and/ or persistent diarrhea (PD) not responding to conventional therapy. DESIGN: Cross-sectional study. PLACE AND DURATION OF STUDY: Gastroenterology Unit of Sindh Institute of Urology and Transplantation (SIUT), from January 2002 to January 2004. PATIENTS AND METHODS: Forty nine children and adolescents with PD (defined as diarrhea greater than 14 days duration) and / or FTT (based on anthropometrical indicators, i.e. weight for length below the 5th centile) were included in the study. Demographic data, weaning practices, breast feeding and family history of CD were documented. Laboratory workup included tissue transglutaminase antibody IgA (tTGA), IgA and IgG antigliadin antibodies (IgA AGA and IgG AGA), HLA typing, upper gastrointestinal endoscopy (EGD) with distal duodenal biopsy (where allowed by parents) and anthropometric data. CD diagnosis was in accordance with the "Guidelines for the diagnosis of and treatment of CD for children" by NASPGHAN. Mann-Whitney U-test, Chi-square test and Fisher's exact test were used for analysis as applicable. RESULTS: Forty nine patients (25 [51%] males) with FTT and or PD were included. The mean (+/-SD) age, height, weight and BMI were: 10.1+/-6.2 years (range, 1-20 years), 107.0 +/- 31.7 cm (45-180 cm), 19.04 +/- 12.5 kg (3-68 kg) and 16.12+/-12.58 kg/cm2 respectively. FTT was present in 30 (61%) patients. Thirty four (69%) children had a history of PD, 38 (77%) had a significant history of weight loss and 32 (65%) children had short stature. Majority of the children (88%) were breast fed. Weaning was started at 6 months of age in 40% and included mixed diet according to age. Protuberant abdomen was present in 26 (53%). Elevated tTGA was significantly (p < 0.001) more frequent in patients with CD. HLA-DQ2 and DQ8 haplotypes were positive in 18 (60%) of these patients. Thirty (61%) patients were positive for CD based on Marsh criteria. CD diagnosis was supported by positive tTGA and/ or AGA tests. Majority i.e. 28/30 patients had Marsh stage 3 disease, while 22/30 patients with CD also had PD. Four CD patients were found to suffer from protein calorie malnutrition (PCM). CONCLUSION: Children who fail to thrive with or without PD may have CD. A positive tTGA test in these children is a useful aid in making a CD diagnosis. Majority of CD positive children were found to carry HLA DQ2 or DQ8 genes. Thus, HLA typing could be used to determine genetic disposition to CD. Majority of FTT had Marsh stage 3 changes on histopathology.

HIV risk in Karachi and Lahore, Pakistan: an emerging epidemic in injecting and commercial sex networks.

The objective of this study was to measure HIV prevalence and risk behaviour in injecting drug users (IDUs), male sex workers (MSWs), Hijras (transgenders), female sex workers (FSWs) and male truckers in Karachi and Lahore, Pakistan. The design was a linked-anonymous cross-sectional study of individuals identified at key venues or through peer referral. Approximately 400 respondents in each group (200 for Hijras) responded to a standardized questionnaire and were tested for HIV antibodies at each site. In Karachi, 23% of IDUs and 4% of MSWs were HIV positive, and HIV-positive individuals were identified in all risk groups in at least one city. Two-thirds of all IDUs used a shared needle in the previous week, and unprotected commercial sex activity with men and women was high. The HIV epidemic has entered IDU and male and female commercial sex networks in Karachi and Lahore. Targeted intervention services must be scaled up and risk group surveillance intensified.

Relationship of HLA antigens and cryoglobulinaemia in hepatitis C virus infected patients.

OBJECTIVE: To find the relationship between human leukocyte antigens (HLA) and cryoglobulin positivity in hepatitis C virus (HCV) infected individuals. METHODS: Eligible individuals selected from pre and post renal transplant settings were divided into three groups. Group A (n = 301) consisted of normal controls, while group B (n = 200) comprised of pathological controls that were HCV antibody (anti-HCV) positive but negative for cryoglobulins. Group C comprised of 56 anti-HCV positive, cryoglobulin positive patients. HLA-A, -B and -DRB1 loci were typed by polymerase chain reaction (PCR) method and relationship between HLA antigens, anti-HCV status and cryoglobulinaemia was analyzed. RESULTS: HLA-A*02, -B*57 and -DRB1*03 were more frequently found among group C members as compared to groups A and B. Only HLA-B* 57 occurrence reached statistical significance (14.3% versus 6% and 4%, corrected P-value = 0.045 and 0.012 and OR = 2.6 and 4 respectively) No differences in the distribution of HLA antigens were seen among healthy and pathological controls. CONCLUSION: The presence of HLA-B*57 confers susceptibility to cryoglobulinaemia in HCV infected patients in our population. HCV positive renal transplant recipients with these alleles should be monitored for cryoglobulin formation).

Cryoglobulinaemia and autoimmune markers in hepatitis C virus infected patients on renal replacement therapy.

OBJECTIVE: To investigate the association of cryoglobulinaemia and autoimmune markers with hepatitis C virus (HCV) infection in patients on maintenance haemodialysis (HD) and post renal transplantation. METHODS: Serum samples of 103 HCV-antibody (anti-HCV) positive and 105 anti-HCV negative patients were investigated for cryoglobulins. These comprised 136 patients on HD and 72 renal transplant recipients. Serum creatinine and liver function tests were obtained on all patients. Rheumatoid factor (RF), anti nuclear antibodies (ANA), anti smooth muscle antibodies (ASMA), liver kidney microsomal antibodies (LKM), immunoglobulins (Igs) and complement levels were performed on all cryoglobulin positive (cryopositive) samples. HCV RNA and genotyping detection tests were done for cryopositive patients. RESULTS: The prevalence of cryoglobulins in patients on HD or after renal transplantation was found to be higher (57.6%) among anti-HCV positive patients compared to the anti-HCV negative patients (42.4%) (P=0.000). RF, ANA and ASMA were also higher in cryopositive HCV infected patients. HCV RNA was present in 84.2% of anti-HCV positive patients. Cryoprecipitable RF activity was found in a higher number of symptomatic patients with HCV genotype 1 compared to HCV genotype 3. CONCLUSION: There is an association of cryoglobulinaemia and autoimmune markers in HCV infected patients on HD, and in HCV positive renal transplant recipients. Also HCV genotype 1 is associated with symptomatic mixed cryoglobulinaemia.

HELICOBACTER PYLORI, HEPATITIS VIRUSES A, C, E, ANTIBODIES AND HBsAg - PREVALENCE AND ASSOCIATED RISK FACTORS IN PEDIATRIC COMMUNITIES OF KARACHI.

OBJECTIVE: To document the prevalence of Helicobacter pylori (H. pylori), Hepatitis A virus (HAV), Hepatitis C virus (HCV), Hepatitis E virus (HEV) antibodies and Hepatitis B virus surface antigen (HBsAg), in the pediatric age group of low socioeconomic urban communities of Karachi and to identify risk factors associated with these infections. DESIGN: Cross-sectional survey. PLACE AND DURATION OF STUDY: Three selected squatter settlements of Karachi during April 2002 to December 2004. PATIENTS AND METHODS: Three hundred and eighty children, ages 5 months to 15 years were investigated. Venous blood samples were collected and questionnaire filled on sociodemographic characteristics (family income, number of dependents in the family, area of living, number of people per room per house, and number of children sharing bed with parents and siblings). Gastrointestinal symptoms were recorded. Anti-HAV IgG (Hepatitis A virus IgG antibody), anti-HCV (Hepatitis C virus antibody), anti-HEV (Hepatitis E antibodies) and HBsAg, were analyzed by enzyme immunoassays (EIAs). Samples were also screened for anti-HIV1/2 (human immunodeficiency virus 1 and 2 antibodies by EIA. IgG antibodies against H. pylori were detected by immunochromatography. RESULTS: A correlation between increasing age and seroconversion was seen for hepatotropic viruses. At 14 years and above,100% of the children were found to be positive for anti-HAV, 26% for anti-HEV, and 1.4%, for anti-HCV while HBsAg was positive in 1.9%. H. pylori infection did not show a significant increase with age. Both anti-HAV and anti-H. pylori were present simultaneously in 30% of the population investigated. CONCLUSION: With age, increasing number of children acquired antibodies against hepatotropic viruses and H. pylori. Occurrence of HBsAg and anti-HEV at a later age suggests horizontal, rather than vertical transmission.

Genetic variability in the precore and core promoter regions of hepatitis B virus strains in Karachi.

BACKGROUND: Hepatitis B virus (HBV) genotypes have distinct geographic distribution. Moreover, much genetic variability has been described in the precore (PC) and basal core promoter (BCP) regions of the HBV genome. The local prevalence of HBV genotypes and mutations has not been well studied. The aim of the present study is to determine the prevalence of HBV genotypes and mutations in the PC and BCP region in HBV strains in Karachi. METHODS: A total of 109 chronic hepatitis B patients with detectable HBV DNA by a PCR assay were enrolled in the study. Sera were tested for HBeAg, anti-HBe antibody and liver profile. HBV genotypes and mutations in the PC and BCP regions were detected by INNO-LiPA line-probe assays. RESULTS: Of the 109 patients investigated, 38 (35%) were HBeAg positive while 71 (65%) were HBeAg negative. Genotype D was present in 100% of the patients. Two patients had co-infection with genotype A. There was no significant difference in the baseline characteristics, mean ALT levels, and presence of clinical cirrhosis in patients with HBeAg positive or negative strains with or without PC and BCP mutations. Of the 38 HBeAg positive patients, 9 (24%) had PC and BCP mutations. In the HBeAg negative patient group, mutations were detected in 44 (62%) of the strains investigated. More than one mutation was common, seen in 26 (37%) patients with HBeAg negative disease and 6 (16%) patients with HBeAg positive disease. Twelve (17%) HBeAg negative patients had dual T1762 and A1764 mutations. None of the HBeAg positive patients had T1762 mutation. Mutations were undetectable in 27 (38%) of patients with HBeAg negative disease. CONCLUSION: Our study shows that type D is the main HBV genotype in Karachi, Pakistan. Significant numbers of patients infected with this genotype have PC and BCP variants. Mutations at more than one site are common. Patients harboring these mutants do not differ significantly in their clinical presentation from patients having wild type infection.

Renal transplantation in developing countries.

Healthcare in developing countries less funded than developed nations (0.8 to 4% vs. 10 to 15%, respectively), and must contend against approximately 1/3 of the population living below the poverty line ($1US/day), poor literacy (58% males/29% females), and less access to potable water and basic sanitation. Cultural and societal constraints combine with these economic obstacles to translate into poor transplantation activity. Donor shortage is a universal problem. Paid donation comprises 50% of all transplants in Pakistan. Post-transplant infections are a major problem in developing countries, with 15% developing tuberculosis, 30% cytomegalovirus, and nearly 50% bacterial infections. The solutions to these problems may seem simplistic: alleviate poverty, educate the general population, and expand the transplant programs in public sector hospitals where commerce is less likely to play a major role. The SIUT model of funding in a community-government partnership has increased the number of transplantations and patient and organ survival substantially. Over the last 15 years, it has operated by complete financial transparency, public audit and accountability. The scheme has proven effective and currently 110 transplants/year are performed, with free after care and immunosuppressive drugs. Confidence has been built in the community, with strong donations of money, equipment and medicines. We believe this model could be sustained in other developing nations.