Impact of different starting doses of atorvastatin on reaching target low density lipoprotein cholesterol levels and health related quality of life in dyslipidemic patients.

Objectives : 1. To compare the percentage of patients that reach target LDL-C goals with 10 mg Vs. 20 mg of atorvastatin as a starting dose. 2. To compare Health Related Quality of Life (HRQOL) in patients on 10 mg Vs. 20 mg of atorvastatin. Methods : A prospective, double blind, parallel groups, unicentric study of patients of dyslipidemia, randomized to receive atorvastatin 10 mg (n=75) or atorvastatin 20 mg (n=75) once daily for 12 weeks. Safety reporting of incidence of adverse events was done. Results : Significantly more number of patients (P<0.05) reached target LDL-C levels at the end of 12 weeks in the 20 mg group (77.27% in the high risk group, 100% in moderately high risk group and 100% in the moderate risk group) when compared to 10 mg group (32% in the high risk group, 75% in moderately high risk group and 83.33% in the moderate risk group). Increase in HRQOL at the end of 12 weeks was also significantly greater (P<0.001) in 20 mg group (27.89%) vs. 10 mg group (19.26%). Conclusions : Selecting the starting dose of atorvastatin according to the patients risk category (by using the Framingham’s algorithm for calculating cardiovascular risk) and the percentage reduction in LDL required, will result in greater success in achieving LDL goals and better quality of life.

Clinical relevance of HBV DNA load in patients with chronic hepatitis B infection.

BACKGROUND: Hepatitis B virus (HBV) DNA detection and quantification are now playing an increasing role in the assessment of disease activity and response to therapy. However, viraemia levels which define various stages of HBV infection have not yet been established. AIM: To define viraemia levels which describe various stages of chronic hepatitis B virus infection. METHODS: In a retrospective study, stored sera samples of chronic hepatitis B virus (CHB) infected patients registered at AIIMS liver clinic, from January 1996 to June 2005 were subjected to competitive, quantitative PCR analysis. RESULTS: The median HBV DNA load was lowest among carriers and highest among patients with chronic hepatitis B [0 (0-8) vs. 7 (0-12) log10 copies/ml, respectively; p<0.05]. As compared to chronic hepatitis patients the DNA load was also lower among cirrhotics [7 (0-12) vs. 4.5 (0-8) log10 copies/ml, respectively; p<0.05] and hepatocellular cancer patients [ 7(0-12) vs. 0 (0-8) log10 copies/ml, respectively; p<0.05]. Patients with carriers had a DNA load which was significantly lower than e antigen negative CHB [0 (0-8) vs. 6 (0-10) log10 copies/ml; p<0.05] or e antigen positive CHB [0 (0-8) vs 8 (0-12) log10 copies/ml; p<0.05]. A threshold of 3.5 log10 copies/ml had sensitivity and specificity of 83% and 58% respectively in differentiating carriers from e antigen negative CHB. There was a strong positive correlation of HBV DNA load with inflammatory grade (R=0.334; p=0.0001), fibrosis stage (R=0.276; p=0.001) and ALT levels (R=0.378; p=0.0001). 82% (9/11) of those who lost e antigen had a decline in HBV DNA levels to <5 log10 copies/ml, whereas only 12.5% (1/8) of those who did not lose e antigen had a decline in DNA load below this level. CONCLUSIONS: HBV DNA viraemia levels correlate positively with the inflammatory grade, fibrosis stage and ALT levels. Most patients who loose e antigen have a decline in DNA load to below 5 log10 copies/ml. Further prospective studies employing repeated measurements are required to define a threshold to differentiate between HBV carriers and e antigen negative CHB.

Characterization of Streptococcus pneumoniae ophthalmic, systemic & commensal isolates by pulsed-field gel electrophoresis & ribotyping.

BACKGROUND & OBJECTIVE: Streptococcus pneumoniae is common in ocular and systemic infections and is a part of normal nasopharyngeal flora. Very few studies regarding genetic analysis of S. pneumoniae isolates causing eye infections are available. This study was undertaken to do pulse field gel electrophoresis (PFGE) analysis and ribotyping of S. pneumoniae isolates obtained from eye infections, systemic infections and nasopharyngeal flora. METHODS: Sixty one well characterized S. pneumoniae isolates (38 from ophthalmic infections, 9 from systemic infections and 14 commensals) were characterized using PFGE of the whole genome after SmaI, restriction enzyme digestion and conventional ribotyping using Escherichia coli rRNA operon as the probe. Phylogenetic tree was drawn using unweighted pair group method analysis (UPGMA). RESULTS: The 38 S. pneumoniae isolates from eye infections belonging to 15 serotypes were placed in to 11 PFGE types and 15 ribotypes. The 9 systemic isolates (7 seotypes) were distributed in 7 PFGE types and 6 ribotypes. The 14 commensal isolates were placed in 11 serotypes, 5 PFGE types and 6 ribotypes. Most of the PFGE types and ribotypes consisting of ocular isolates also contained systemic and commensal isolates. INTERPRETATION & CONCLUSION: Considerable genetic similarity was observed between the isolates from ocular and systemic infections and those colonized in nasopharynx. PFGE analysis could differentiate majority of the isolates according to site of infections. There was a considerable DNA polymorphism within the studied bacterial population.

Viral hepatitis in India.

Viral hepatitis is a major public health problem in India, which is hyperendemic for HAV and HEV. Seroprevalence studies reveal that 90%-100% of the population acquires anti-HAV antibody and becomes immune by adolescence. Many epidemics of HEV have been reported from India. HAV related liver disease is uncommon in India and occurs mainly in children. HEV is also the major cause of sporadic adult acute viral hepatitis and ALF. Pregnant women and patients with CLD constitute the high risk groups to contract HEV infection, and HEV-induced mortality among them is substantial, which underlines the need for preventive measures for such groups. Children with HAV and HEV coinfection are prone to develop ALF. India has intermediate HBV endemicity, with a carrier frequency of 2%-4%. HBV is the major cause of CLD and HCC. Chronic HBV infection in India is acquired in childhood, presumably before 5 years of age, through horizontal transmission. Vertical transmission of HBV in India is considered to be infrequent. Inclusion of HBV vaccination in the expanded programme of immunization is essential to reduce the HBV carrier frequency and disease burden. HBV genotypes A and D are prevalent in India, which are similar to the HBV genotypes in the West. HCV infection in India has a population prevalence of around 1%, and occurs predominantly through transfusion and the use of unsterile glass syringes. HCV genotypes 3 and 2 are prevalent in 60%-80% of the population and they respond well to a combination of interferon and ribavirin. About 10%-15% of CLD and HCC are associated with HCV infection in India. HCV infection is also a major cause of post-transfusion hepatitis. HDV infection is infrequent in India and is present about 5%-10% of patients with HBV-related liver disease. HCC appears to be less common in India than would be expected from the prevalence rates of HBV and HCV. The high disease burden of viral hepatitis and related CLD in India, calls for the setting up of a hepatitis registry and formulation of government-supported prevention and control strategies.

Serotype distribution of Streptococcus pneumoniae isolates from ophthalmic and systemic infections and of commensal origin.

BACKGROUND AND OBJECTIVES: Information regarding serotype distributions of Streptococcus pneumoniae causing ophthalmic infections is scanty. This study was therefore undertaken to determine the antimicrobial susceptibility status and serotypes of S. pneumoniae isolated from various ophthalmic infections and to compare with those isolated from systemic infections and commensal nasopharyngeal flora. METHODS: Thirty eight of S. pneumoniae isolates from ophthalmic infections, 9 from systemic infections and 14 from the nasopharynx of apparently healthy school children were biochemically characterized and tested for in vitro antimicrobial susceptibility to various antibiotics. Serotyping of these 61 isolates was done by a rapid co-agglutination method. RESULTS: All the 61 isolates were sensitive to oxacillin (penicillin) and susceptibility against other antimicrobials was variable. No multidrug resistance was observed. The 38 ophthalmic isolates were distributed in 15 different serotypes. Most prevalent serotypes were 14, followed by 8 and 19F. The 9 systemic and 14 commensal. isolates of S. pneumoniae were distributed in 7 and 11 serotypes respectively. Three of the systemic and six of the commensal serotypes were observed in ophthalmic infections whereas four of the commensal serotypes were observed in systemic infections. INTERPRETATION AND CONCLUSION: Resistance to penicillin was not observed. In ophthalmic infections, a wide range of serotypes of S. pneumoniae were observed. More than half of the commensal serotypes obtained in the study as well as majority of the systemic serotypes were observed in ophthalmic infections.

Vitamin E-based therapy is effective in ameliorating transaminasemia in nonalcoholic fatty liver disease.

BACKGROUND: Comparative trials of ursodeoxycholic acid (UDCA), vitamin E and weight management programs among patients with nonalcoholic fatty liver disease (NAFLD) are lacking. AIM: To find an effective single agent or combination of agents for management of NAFLD. METHODS: In this retrospective study, consecutive patient with histologically confirmed NAFLD with raised ALT were included. The patients received either weight management (exercise and therapeutic lifestyle changes [TLC] diet with a target to reduce body weight 10% in 6 months) (group I) ; weight management + UDCA (300 mg BID) (group II); or weight management + UDCA + vitamin E (400 mg OD) (group III). Outcome measure was normalization of ALT. RESULTS: 42 patients (18, 12 and 12 in groups I, II and III, respectively) were included between 1996 and 2004. All patients in group III normalized their ALT levels, which was significantly higher than numbers in group I (8/18) and group II (5/12); (p=0.003). Post treatment ALT was significantly lower in group III (28.6 [9.3]) as compared to group I (59.3 [32.2]) and group II (49.0[31.8]); (p=0.01). Type of therapy received was the only factor predictive of ALT normalization. CONCLUSION: Combination regimen containing vitamin E appears to be effective in normalizing ALT among NAFLD patients.

Mother-to-child transmission of hepatitis E virus infection.

OBJECTIVES: Water borne or enterically transmitted non-A-non-B hepatitis is a major public health problem in India. Many of these cases carry fatal outcome. The hepatitis E virus (HEV) has been considered to be the most important causative agent of this entity. The severity and fatality rates of HEV infection are reported to be rather more in pregnant women. However, there is meager information from India, on mother to child transmission of this agent. METHODS: During 1997-98, we studied 60 pregnant women suspected to have acute viral hepatitis to understand the frequency of various viral etiologies, disease course and outcome of the pregnancy. Six cord blood samples were tested for IgG, and IgM antibodies against hepatropic viral agents and also for hepatitis E virus RNA by RT-nested PCR using ORF-1 as target. RESULTS: Of the 60 pregnant patients hospitalised at All India Institute of Medical Sciences, New Delhi for acute hepatitis, 22 (37%) were positive for IgM anti-HEV antibodies and 10% were infected with hepatitis B virus. Co-infection of HEV with Hepatitis B and C was seen in 1 and 2 patents, respectively. Most (72%) of the HEV infected patients were in third trimester of pregnancy (P<0.05). Of the 6 cord blood samples tested 3 (50%) were positive for HEV RNA. Though, all mothers were RNA positive, half of the babies did not get infected in utero with HEV. Fourteen of the 22 (63.6%) HEV infected mothers developed fulminant hepatic failure and all died. CONCLUSION: The mortality rate in HEV [corrected] infected mothers was 100%. Mother to child transmission of hepatitis E virus infection was established in 50%.

Vaccination for hepatitis A virus is not required for patients with chronic liver disease in India.

BACKGROUND: Hepatitis A virus (HAV) vaccination is recommended worldwide for patients with chronic liver disease to prevent decompensation due to superinfection with HAV. India being endemic for HAV, the prevalence of pre-existing antibodies against HAV due to subclinical exposure to the virus in childhood among patients with chronic liver disease may be high and, therefore, vaccination may not be needed. However, data are lacking on the prevalence of HAV antibody among patients with chronic liver disease in India. METHODS: Two hundred fifty-four patients attending the Liver Clinic at the All India Institute of Medical Sciences, New Delhi during the past 5 years and diagnosed to have either chronic hepatitis due to the hepatitis B virus (n = 76), hepatitis C virus (n = 84) or cirrhosis of the liver due to the hepatitis B (n = 47) or C (n = 47) virus were tested for the presence of IgG anti-HAV antibody in their sera (using a commercial ELISA kit). RESULTS: Two hundred forty-eight (97.6%) patients tested positive for IgG anti-HAV. The prevalence of anti-HAV positivity was similar among patients with chronic hepatitis B (74, 97.4%), chronic hepatitis C (82, 97.6%), cirrhosis of the liver due to the hepatitis B (46, 97.8%) and hepatitis C (46, 97.8%) virus. CONCLUSION: Vaccination against HAV is not required among patients with chronic liver disease in India as there is a very high prevalence of pre-existing antibodies in these patients.

Sero-epidemiology of hepatitis E virus (HEV) in urban and rural children of North India.

OBJECTIVE: To estimate the prevalence of anti-HEV IgG and IgM antibodies to ORF3 peptide of Hepatitis E virus genome in an age stratified urban and rural population of children. DESIGN: Cross sectional survey. SETTING: Pediatric out-patient clinics in a tertiary hospital and a rural dispensary. METHODS: Study subjects between 6 months and 10 years with minor, non-hepatic illnesses were recruited for the study from March to December 1996. Baseline demographic details, drinking water source, sewage disposal methods, reasons for attending the hospital, histories of parenteral exposure in the past 12 months and acute hepatitis in the subjects and the family in the previous six months were obtained. Serum anti-HEV IgG antibodies were screened in all subjects, and in those who were positive, anti-HEV IgM antibodies were assayed as an indicator of recent infection. Serum aminotransferase (ALT) was estimated in those who were anti-HEV IgM antibody positive. RESULT: Out of 2160 subjects recruited, 2070 samples could be screened for anti-HEV IgG antibodies. In the urban population (n = 1065) anti-HEV IgG antibodies were detected in 306 subjects (28.7%; 95% CI 26.0-31.6) and of these 131 (42.8%; 95%CI 37.2-48.6) were anti-HEV IgM antibody positive. Amongst 1005 rural children, anti-HEV IgG antibodies were present in 239 (23.8%; 95% CI 21.1-26.4) and IgM antibodies in 113 (47.3%; 95% CI 40.9-53.7) children. The antibodies were present since the first year of age till 10 years of age and, increased with advancing age. Serum transaminases were raised in 7.5% (9/120) and 5.5% (5/88) of subjects with anti-HEV IgM antibodies in urban and rural centers respectively. Overall the seroprevalence of IgG antibodies against HEV were significantly more in urban as compared to that in rural subjects (p = 0.011). However, proportion of children with anti-HEV IgG carrying IgM antibodies was similar in the two study groups (p = 0.298). A model for estimating expected prevalence of anti-HEV IgG antibodies was developed. The observed antibody prevalence in both urban and rural subjects at each age interval after 48 months was less as compared to the expected levels and this gap increased with advancing age categories. It appeared that there was a decay of HEV antibodies with time. CONCLUSIONS: Children are susceptible to HEV infection since early infancy. The probability of exposure to HEV during childhood was higher in urban than rural population. Seropositivity to HEV antibodies increased by over 2 times beyond 4 years of age as compared to younger age. Anti-HEV IgG antibodies appear to wean off with increasing age.

Diagnosis of hepatitis C virus infection by ELISA, RIBA and RT-PCR: a comparative evaluation.

OBJECTIVES: To evaluate the efficacy of second-generation ELISA (ELISA-2), third-generation ELISA (ELISA-3) and third-generation recombinant immunoblot assay (RIBA 3.0) for detection of antibodies to hepatitis C virus (anti-HCV) in comparison with reverse transcriptase-polymerase chain reaction (RT-PCR) to detect HCV RNA for the diagnosis of hepatitis C. METHODS: Sera of 108 patients with chronic liver disease (CLD) were analyzed by ELISA-2, ELISA-3, RIBA 3.0 and RT-PCR in the first part of the study; in the second part, sera of 105 patients with non-chronic liver disease were evaluated with ELISA-3, RIBA 3.0 and RT-PCR. RESULTS: In the CLD group, anti-HCV was positive in 4.6%, 14.8% and 16.6% by ELISA-2, ELISA-3 and RIBA 3.0, respectively. Among these anti-HCV positive cases, HCV RNA was positive in 100%, 58.9% and 64%, respectively. ELISA-2 did not give false-positive results, but missed substantial number of anti-HCV positive cases (p < 0.001). In the second group, anti-HCV was positive in 76.3% by ELISA-3 and 68.6% by RIBA 3.0 (p:ns). HCV-RNA was positive in 88.7% of ELISA- and RIBA-positive cases; in 60% of ELISA-positive, RIBA-indeterminate cases; and in 46.4% of ELISA-negative, RIBA-negative cases. CONCLUSIONS: ELISA-2 is not a suitable assay for routine screening. ELISA-3 was at par with RIBA 3.0 and it can be recommended for routine screening for anti-HCV. RT-PCR for HCV is of value in detecting early viremic, anti-HCV negative cases; this may be of importance in the treatment of hepatitis C.

Evaluation of third generation anti-HCV enzyme immunoassays.

The Hepatitis C Virus (HCV) is a major cause of post transfusion hepatitis. The introduction of HCV antibody screening has reduced the risk of post transfusion hepatitis significantly. However, the test is yet to be used routinely in blood banks of several developing countries with limited resources. We have developed an Enzyme immunoassay using synthetic peptides. The test was compared to seven commercial tests available in the Indian market. The test was evaluated using a panel of 90 sera which were chosen from an earlier panel based on detection of HCV RNA by Reverse Transcription Polymerase Chain Reaction RT-PCR. In case of any discrepancy the sera were further analysed by Line immunoassay (LIA). The sensitivity of the in house EIA was 90%. The specificity of the commercial EIAs varied.

Development of an antigen capture enzyme immuno assay using genus specific monoclonal antibodies for detection of Chlamydia in clinical specimens.

An antigen capture enzyme immuno assay (EIA) for Chlamydia antigen detection was developed using polyclonal rabbit immunoglobulins against C. trachomatis, a genus specific antichlamydial murine monoclonal antibody (IgG) against major outer membrane protein (MOMP) antigen of C. trachomatis and a commercial anti mouse IgG immunoglobulin conjugated with HRPO. The test was evaluated against a direct immunofluorescence assay (DFA). Conjunctival specimens from 178 patients with follicular conjunctivitis and cervical specimens from 82 patients with cervicitis were tested for Chlamydia antigen detection by both the tests. Chlamydia antigen was detected in 69/178 (38.76%) and 68/178 (38.20%) of the conjunctival specimen by using EIA and DFA tests respectively. It could also be detected in 24/82 (29.27%) and 22/82 (26.83%) of the cervical specimens by EIA and DFA tests respectively. The sensitivity of the EIA test was 92.64 per cent and 86.36 per cent for the conjunctival and cervical specimens respectively against the reference DFA test. The specificity of the EIA test was found to be 94.54 and 91.66 per cent respectively against the reference DFA test.

The epidemiologic significance and clinical pattern of HCV induced chronic hepatitis in India.

Hepatitis C virus (HCV) ribonucleic acid (RNA) was tested for in a group of 16 defined non-B chronic hepatitis patients using specific reverse transcription polymerase chain reaction (RT-PCR). These were chosen from amongst 56 biopsy proven cases of chronic hepatitis of which majority (40) were positive for hepatitis B virus infection. Hepatitis C virus RNA could be demonstrated in 12 (75%) of remaining 16 cases. These include all seven patients positive for antibody to HCV. Two of these patients had past history of blood transfusion and in another two the clinical course started with severe acute liver disease. This study establishes the association of HCV with severe liver disease. The clinical and biochemical profiles are also discussed. In view of limited sensitivity of the antibody assays it is justified to develop diagnostic testes based on local strains.

Development & use of antichlamydial monoclonal antibodies for rapid diagnosis of follicular conjunctivitis.

We developed four independent murine hybrid clones producing IgG class and genus specific monoclonal antibodies against the major outer membrane protein (MOMP) of Chlamydia trachomatis, L2 serovar. All antibodies reacted with a single epitope of MOMP. In indirect immunofluorescence assay using one of the antibodies chlamydiae could be detected in 12 of 23 conjunctival scrapings from patients of follicular conjunctivitis. Giemsa staining could detect classical inclusion bodies in only 5 of these 23 smears. Antigen detection using these monoclonal antibodies may be used as a rapid diagnostic test in clinical and epidemiological screening for chlamydial infection.

Characterisation of the Indian strain of duck hepatitis B virus (DHBV) and development of a carrier duck colony for antiviral drug testing.

Duck hepatitis B virus (DHBV) infected carrier ducks serve as a useful model for testing anti-hepadnavirus drugs. This needs a well characterised duck hepatitis B virus strain. We cloned and sequenced the complete genome of duck hepatitis B virus strain of Indian origin. It was 3021 nucleotides in length and had all the recognisable open reading frames (Polymerase: 20-2530 nucletides, Surface: 693-1787 nucleotides and Core: 2518-412). Using an inoculum of 50 microliters serum containing 1 x 10(11) virus particles/ml, we could infect 80% of one day old ducklings and develop a duck colony.

Chronic hepatitis in a large Indian hospital.

BACKGROUND. In developed countries as well as in Southeast Asia, the hepatitis B and C viruses are the main causes of chronic hepatitis. In India, however, there have been no major investigations on the aetiology of chronic hepatitis. (The hepatitis E virus which is responsible for half the sporadic and most of the epidemic cases of acute viral hepatitis in India does not cause chronic disease.) We, therefore, studied the profile of chronic hepatitis in India. METHODS. The clinical presentation, aetiology, serology and histological changes were studied prospectively in 48 patients with chronic hepatitis admitted to the All India Institute of Medical Sciences, New Delhi. Of these, 44 (92%) had chronic active hepatitis, 3 (6.3%) had chronic persistent hepatitis and 1 (2%) had chronic lobular hepatitis. RESULTS. The hepatitis B virus was the aetiological agent in 24 (50%) of these patients, the hepatitis D virus in association with hepatitis B virus in 10 (21%), the hepatitis C virus in 7 (15%) and the non-A, non-B viruses other than the hepatitis C virus in 6 (13%). One patient (2.0%) had autoimmune chronic active hepatitis. Jaundice at presentation was seen in 33 (69%) patients and more than half had hypoalbuminaemia (< 3 g/dl) with a prolonged prothrombin time. Alanine aminotransferase levels were less than 5 times above normal in over two-thirds of the patients. The highest alanine aminotransferase values were observed in patients with hepatitis D virus infection whereas the lowest were seen in patients with non-A, non-B related chronic active hepatitis. Histological examination revealed bridging necrosis in 40 (91%) patients with chronic active hepatitis indicating a severe form of disease. Replication of the hepatitis B virus was seen in 13 patients with chronic hepatitis, 5 of whom had hepatitis D virus-induced chronic hepatitis. Patients with hepatitis B virus replication had higher alanine aminotransferase values and more severe bridging necrosis than patients who did not have replicating viruses. Higher alanine aminotransferase values, ascites and oesophageal varices were encountered more frequently in patients with hepatitis B and D virus than in those with non-A, non-B related chronic hepatitis. CONCLUSION. Chronic hepatitis is not uncommon in India. It presents with evidence of severe disease and, as elsewhere, is most frequently caused by the hepatitis B virus.