A study of impact of cost-effective nutritional supplement in patients on maintenance hemodialysis.

Nutritional status in patients on hemodialysis is always of concern as malnutrition predisposes to excess morbidity and mortality. Most of the nutritional supplements available in the market are expensive. We explored the possibility of improving nutrition of the patients on maintenance hemodialysis by supplementation of calories and proteins that can be given in the form of a palatable and economical gruel in this prospectively designed, open labeled study. Patients who were on maintenance hemodialysis (twice a week) for a period of at least 6 months were divided into two groups. The study group was given the gruel supplement and the control group was not given the gruel supplement. Nutritional status was assessed in the study group and controls at 0 and 3 months by the following parameters: percentage body fat, mid arm muscle circumference and serum albumin. Analysis of results revealed that there was a significant decline in the protein intake at the end of the 3(rd) month in the control group (P = 0.01). Other parameters did not show significant change at the end of the study period in both groups. The nutritional supplement can be assumed to have helped at least in the maintenance of protein intake over this short period and could possibly in the long run contribute to improvement of nutritional parameters.

Non-expression of HLA-C*07:55N is caused by a premature stop codon in exon 3.

Sequencing of HLA-C*07:55N assessed the presence of a premature TAG stop codon at residue 113.

A hospital-based study of splenomegaly with special reference to the group of indeterminate origin.

In any study there remains a proportion of cases, about 25-40%, where cause of splenomegaly is not identified on usual evaluation, that is labelled as indeterminate group. The aim of this study was to evaluate various causes of splenomegaly. Thereafter the patients with splenomegaly of indeterminate origin were to be re-evaluated with detailed investigations (for the cause of splenomegaly). Causes of splenomegaly were looked into 100 adult patients with splenomegaly, admitted over a period of ten months in a teaching hospital in South India. Patients having ascites were excluded from the study. Malaria was the commonest cause of splenomegaly, observed in 22 patients. Other causes, in order of importance, were chronic myeloid leukaemia (n=11), non-cirrhotic portal fibrosis (n=9), enteric fever (n=9), cirrhosis of liver (n=8) and hyper-reactive malarial splenomegaly also called as tropical splenomegaly syndrome (n=7) and so on. Hyper-reactive malarial splenomegaly was the commonest cause (7 of 24 patients) of massive splenomegaly. Twenty-three patients had splenomegaly of indeterminate origin ie, cause could not be detected on first assessment. Detailed re-evaluation with repeat investigations including liver biopsy revealed the causes as follows: Hyper-reactive malarial splenomegaly -7 (30.4%), non-cirrhotic portal fibrosis - 4 (17.4%), cirrhosis of liver - 4 (17.4%) and iron deficiency anaemia - 5 (21.7%). In 3 patients (13.0%), no diagnosis could be arrived at despite best efforts. Obscure splenomegalies may be due to conditions like hyper-reactive malarial splenomegaly, non-cirrhotic portal fibrosis, iron deficiency anaemia, and even cirrhosis of liver, while malaria is still the most important cause of splenomegaly in India. Whereas the overall incidence of hyper-reactive malarial splenomegaly was only 7% in this study, it stood as the leading cause (29.2%), when analysed among patients with massive splenomegaly. Liver biopsy should be performed in all cases of obscure splenomegaly to arrive at the final diagnosis.

Nosocomial infections due to Acinetobacter species: Clinical findings, risk and prognostic factors.

PURPOSE: Nosocomial infections caused by Acinetobacter species is of increasing concern in critically ill patients, and the risk factors for this infection are not well established. The present investigation was done to determine incidence of nosocomial Acinetobacter infections. Our study retrospectively attempts to find risk and prognostic factors for the nosocomial acquisition of Acinetobacter infection. METHODS: The medical records of 43 patients with Acinetobacter infection during two-year period (Oct1998-Oct2000) were reviewed to find the factors involved in the nosocomial acquisition of Acinetobacter. Acinetobacter isolates that were obtained from these patients were phenotypically typed using carbon assimilation tests. Antimicrobial susceptibility testing results were noted from the microbiology records. RESULTS: Acinetobacter baumannii accounted for 41.8% (n=18) of all the infections. By multivariate logistic regression analysis, only resistant antibiotype {(Ceftazidime- OR, 7.13 [95% CI, 1 to 46];p= 0.044); (Cefotaxime- OR, 6.09 [CI, 0.87 to 30];p = 0.045)} and mechanical ventilation (OR, 5.84 [CI, 0.83 to 31];p = 0.05) were found to be potential independent risk factors for mortality. Overall mortality rate was 33%. CONCLUSIONS: Most of A. baumannii isolates were multidrug resistant in our set up and infections due to them were associated with high mortality. Prevention of Multiple drug resistant (MDR) A. baumannii infections was achieved after discontinuation of cefotaxime in ICU. Infection with resistant clones and mechanical ventilation were found to be potential independent risk factors for mortality.

Shell Vial Culture assay for the rapid diagnosis of Japanese encephalitis, West Nile and Dengue-2 viral encephalitis.

BACKGROUND: Encephalitis caused by flaviviruses, Japanese encephalitis virus (JEV) and West Nile virus (WNV) is responsible for significant morbidity and mortality in many endemic countries. Dengue-2 (Den-2) virus is a recent addition to the list of encephalitogenic viruses, after its Central Nervous System (CNS) invasion capability has been established. There is a wide array of laboratory tools that have helped us not only in the diagnosis of these conditions but also in understanding their pathogenesis and pathology. However, there are no reports of Shell Vial Culture (SVC), a centrifuge enhanced tissue culture assay that has revolutionized viral culturing in terms of rapidity and sensitivity being optimized for these flaviviral encephalitic conditions. The present study is an attempt to standardize and evaluate the usefulness of SVC for the laboratory diagnosis of JE, WN and Den-2 encephalitis cases and to compare it with Indirect Immunofluorescence (IIF) technique that detects cell associated virus antigen. Analysis of the various clinical parameters with respect to viral etiology has also been carried out. RESULTS: Pediatric patients constituted the major group involved in the study (92%). Etiological diagnosis of viral encephalitis could be established in twenty nine (58%) patients. JE encephalitis was the commonest with 19 (39%) cases being positive followed by, WN (9 cases-18%) and Den-2 (one case). IIF test could detect antigens of JE, WN and Den-2 viruses in 16(32%), 7(14%) and 1 case respectively. Shell vial culture assay picked up all cases that were positive by IIF test. In addition, SVC assay could detect 3 and 2 more cases of JE and WN encephalitis respectively, that were negative by the IIF test. CONCLUSION: Shell vial culture is a rapid and efficient tool for the etiological diagnosis of JE, WN and Den-2 encephalitis cases. Early, prompt collection, transport and processing of the CSF samples, would make SVC a better method for the rapid diagnosis of these flaviviral infections.

Epidemiological investigation of nosocomial Acinetobacter infections using arbitrarily primed PCR & pulse field gel electrophoresis.

BACKGROUND & OBJECTIVE: Nosocomial infections caused by Acinetobacter spp. are a significant problem worldwide. Information on epidemiological investigation of outbreaks caused by Acinetobacter species in India is lacking. The present investigation was carried out to elucidate molecular epidemiology of Acinetobacter species isolated from nosocomial infections in a tertiary care hospital in south India using two DNA-based typing methods. METHODS: The medical records of 43 patients with Acinetobacter infection during a period of 24 months were reviewed and Acinetobacter isolates obtained from these patients were characterized phenotypically by assimilation tests and genotypically by arbitrarily primed PCR (AP-PCR) and pulse field gel electrophoresis (PFGE). Susceptibility testing results of the Acinetobacter isolates were also analysed. RESULTS: Most of the infections were nosocomial, and the majority of these were acquired in intensive care units (ICUs). A. baumannii accounted for 41.8 per cent (n=18) of all pneumonia acquired in the ICU. AP-PCR with M13 primer distinguished 8 different PCR patterns comprising of 2 to 6 DNA fragments of 0.1 to 1.0 kb. PFGE identified 9 distinct profiles with five subvarients. By APPCR, epidemiologically unrelated strains could not be differentiated and often differences within biotypes of A. baumannii were not detectable. ApaI macrorestriction (PFGE) identified at least 4 outbreaks caused by 3 clones of A. baumannii and one clone of DNA group 13TU, one replacing the other in a well-defined temporal order. INTERPRETATION & CONCLUSION: Most of A. baumannii isolates were multidrug resistant. PFGE was more discriminatory [Discriminatory Index (DI)=0.96 than AP-PCR fingerprinting (DI=0.88)] in the present study. However, AP-PCR fingerprinting is more useful as a simple and rapid identification technique for epidemiological investigation of nosocomial Acinetobacter infections.

Can duodenal ulcer perforation be linked to herpes simplex virus infection?

BACKGROUND: Both Herpes simplex infection and duodenal ulcer recur frequently, tend to remain localized, and show remissions and exacerbations. Published data on a link between the two are contradictory, and there are no data on the association of Herpes simplex infection with perforated duodenal ulcer. METHODS: 187 patients in four groups were studied: group I--controls (n = 35), group II--non-ulcer dyspepsia (n = 35), group III--chronic non-perforated duodenal ulcer (n = 35), and group IV--perforated duodenal ulcer (n = 82). Titers of IgG antibodies against HSV-1 and HSV-2 were determined using enzyme immunoassays. RESULTS: The seropositivity rate for both HSV-1 (80%) and HSV-2 (77%) was high in the control population. Among patients with perforated duodenal ulcer, antibodies against HSV-1 (94%) but not those against HSV-2 (83%), were found more frequently than in groups I and III. HSV-1 seropositivity was significantly higher in patients with a short duration of preperforation symptoms. Absolute titers for both anti-HSV-1 and anti-HSV-2 were higher in patients with perforated duodenal ulcer than in controls and those with chronic non-perforated duodenal ulcer. CONCLUSION: Herpes simplexvirus, especially HSV-1, may have a role in the causation of perforated duodenal ulcers.

Detection of pneumolysin in cerebrospinal fluid for rapid diagnosis of Pneumococcal meningitis.

BACKGROUND & OBJECTIVES: Pneumolysin, a toxin produced by Streptococcus pneumoniae is associated with virulence and is found in all invasive isolates. Its role as a diagnostic tool has recently been exploited. Most of the methods used are based on molecular techniques and are not cost-effective. The present study was undertaken to evaluate a simple, rapid and cost-effective method to detect pneumolysin in CSF as a diagnostic test for pneumococcal meningitis. METHODS: A total of 75 CSF samples from children with presumptive diagnosis of acute pyogenic meningitis or encephalitis were subjected to Gram stain, culture and pneumolysin detection by Cowan 1 staphylococcal protein A co-agglutination technique. RESULTS: Pneumolysin was detected in 26(78.8%) of 33 culture proven CSF samples and 4(9.5%) of 42 culture negative samples. Antigen detection by Co-A had a specificity of 90 per cent and a sensitivity of 79 per cent when compared with culture. Compared to Gram stain, pneumolysin Co-A had a specificity and sensitivity of 91.0 and 92.0 per cent respectively. INTERPRETATION & CONCLUSION: Detection of pneumolysin was found to be a simple, low cost antigen detection assay for rapid diagnosis of pneumococcal meningitis, for routine use in the developing countries.

In vitro susceptibility pattern of Acinetobacter species to commonly used cephalosporins, quinolones, and aminoglycosides.

PURPOSE: Acinetobacter spp. is an emerging important nosocomial pathogen. Clinical isolates of this genus are often resistant to many antibiotics. The in vitro susceptibility of Acinetobacter isolates obtained from patients were tested for currently used antibiotics. In addition, the study aimed at biotyping of Acinetobacter baumannii. METHODS: A total of 66 isolates were phenotypically characterised through a large panel of 25 carbon assimilation tests and susceptibility through disc diffusion method with 10 antimicrobial agents were tested. MICs were determined only for second line broad-spectrum drugs such as cefotaxime, ceftazidime, amikacin, ciprofloxacin, and ofloxacin using NCCLS guidelines. RESULTS: Multiple drug resistance (MDR) was only witnessed in A. baumannii and not in other Acinetobacter species. Aminoglycosides such as amikacin, netilmicin were most active against the MDR isolates tested (60% susceptibility). Ceftazidime was more active than cefotaxime. MDR A. baumannii strains were susceptible only to amikacin, netilmicin and ceftadizime. Ciprofloxacin had poor activity irrespective of isolates belonging to different DNA groups tested (58% resistance overall, 79% among A. baumannii). Strains of Biotypes 6 and 19 of A. baumannii showed broader resistance than those of biotype 10 and others. CONCLUSIONS: Strains of A. baumannii from patients in our hospital, were generally more resistant to quinolones, -lactam antibiotics, first and second generation cephalosporins and partially resistant to third generation cephalosporins and aminoglycosides. The strains belonging to other DNA groups of Acinetobacter were comparatively less resistant than A. baumannii, except ciprofloxacin. This study suggests that, a combination therapy, using a third generation cephalosporin and amikacin, would be best choice for treating Acinetobacter infections.

Learning from an outbreak: ESBL- the essential points.

Extended spectrum beta-lactamase (ESBL) producing strains of Klebsiella pneumoniae have emerged as important nosocomial pathogens. The present report describes an outbreak of ESBL positive K. pneumoniae in a neonatal intensive care unit of a tertiary care center in southern India. The clinical and microbiological significance of multiresistant gram negative ESBL producers have been discussed in light of this outbreak by multi-resistant gram negative bacilli. The review also offers some practical guidance regarding infection control and therapeutic options.

Rational use of anti-snake venom (ASV): trial of various regimens in hemotoxic snake envenomation.

BACKGROUND: Viperine snake bites cause hemotoxicity in the form of coagulation dysfunction. Optimal dose requirement of anti-snake venom (ASV) and duration of therapy in such situation have not yet been fully explored. Our aim in this study was to compare two low-dose continuous infusion regimes with the standard high dose intermittent bolus regime in treating systemic envenomation and preventing its recurrence. METHODS: A prospective interventional study was conducted on 90 adult patients with snake bite with hemotoxicity. Patents were allocated into three treatment regimes, each regime being tried on 30 patients. Regimen I (standard high dose regimen) consisted of conventional, intermittent bolus dosage of 100 ml of ASV as a loading dose followed by 50 ml every six hours till whole blood coagulation time (CT) became normal. Regimen II consisted of 30 ml of ASV as a loading dose followed by 30 ml continuous infusion every six hours till two CTs at an interval of six hours were normal and a further dose of 30ml over 24 hours. Regimen III was similar to Regimen II in all aspects except that loading dose was 70 ml (instead of 30 ml). RESULTS: In patients with mild envenomation, even though the average requirement of ASV was only marginally lower in Regimen II (128.6 ml) as compared to in Regimen I (137.5 ml), one patient on Regimen I had relapse of coagulation dysfunction. In patients with moderate envenomation, average requirement of ASV was 221.3 ml and 179 ml in Regimens II and III respectively, which was much less than in Regimen I (343.8 ml) (p values 0.05 and 0.01 respectively). Further, no patient receiving Regimen III had relapse of coagulation dysfunction. In severe envenomation, average dose of ASV required was almost similar in Regimens II and III, i.e., 213.7 ml and 233.7 ml respectively, as compared to 433.3 ml required in Regimen I (p values 0.02 and 0.001 respectively). However, time lapse for CT normalization was only 18 hours in Regimen III as compared to 23.6 hours and 24 hours in Regimens I and II respectively. CONCLUSION: Regimens consisting of continuous intravenous infusion of ASV i.e., Regimen II in mild envenomation and Regimen III in moderate and severe envenomation are likely to make significant saving of ASV and reduction of recurrence of coagulation dysfunction.

Japanese encephalitis in and around Pondicherry, South India: a clinical appraisal and prognostic indicators for the outcome.

Japanese encephalitis (JE) is numerically one of the most important causes of viral encephalitis worldwide, with an estimated 50,000 cases and 15,000 deaths annually. About one-third of patients die and half of the survivors have severe neuropsychiatric sequelae. Three hundred patients clinically suspected of JE were tested in the present study. Laboratory confirmation of JE was on the basis of detection of antigen or presence of JE-specific IgM antibody and/or neutralizing antibody in a single CSF sample. The risk factors that were associated with fatal outcome were determined. Japanese encephalitis infection was confirmed in 70.7 per cent (212/300) of the patients. All patients were from rural areas and with low socioeconomic background. Prominent clinical findings were: fever in 100 per cent (212/212) patients, altered sensorium in 87.73 per cent (186/212), convulsion in 85.84 per cent (182/212), headache in 50 per cent (106/212), and vomiting in 47.64 per cent (101/212). The final clinical outcome was available for only 68.39 per cent (145/212) of patients, as children were taken home against medical advice. Of these, 35.86 per cent (52) died while 63.44 per cent (92) of patients survived. Correlations of investigative findings with the final outcome revealed that absence of virus-specific IgM and neutralizing antibodies in CSF were associated with fatal outcome. In patients diagnosed with Japanese encephalitis the presence of a virus-specific immune response is associated with a favourable outcome and an important parameter in recovery from illness.

Seroprevalence of Hepatitis C virus in a hospital based general population in south India.

Seroprevalence of Hepatitis C virus (HCV) among hospital based general population was determined using a third generation ELISA. The study population comprised of 661 individuals (including 36 health care workers) attending a tertiary care hospital in Pondicherry, south India. The overall seroprevalence was found to be 4.8% (95% confidence interval [CI]=3.2-6.4%). The seroprevalence in males and females was 5.9% (95% CI=3.5-8.3%) and 3.3% (95% CI= 1.2-5.4%) respectively. There was no statistically significant difference in the proportion of individuals who were positive in case of males and females (p>0.05). None of the health care workers tested positive for antibodies to HCV.

Co-infection with hepatitis C virus and human immunodeficiency virus among patients with sexually transmitted diseases in Pondicherry, South India.

Hepatitis C virus (HCV), which is usually transmitted by blood and blood products is emerging as an important agent in the list of sexually transmitted diseases (STDs). The present study was undertaken to document the burden of HCV infection in individuals with STDs in this tertiary care hospital in South India. One hundred serum samples collected from individuals with STDs were tested for antibodies to HCV by a third generation ELISA. All the samples were also screened for HIV infection. Six out of 100 individuals were found to possess antibodies against HCV (95% confidence interval [CI=1.3-10.7%). Fourteen out of 100 samples were positive for HIV (95% CI=7-20.9%). The seroprevalence of HCV in HIV positive individuals was 21.4% (3/14) whereas the corresponding figure for HIV negative individuals was only 3.5% (3/86). The difference was found to be statistically significant (p<0.01).

Can fall in anti-H. pylori IgG titres indicate eradication?

At present, it is not clear whether a fall in serological titres of anti-H. pylori IgG can be used for confirming eradication. A prospective study was conducted using varying cut-off levels from (10% to 50%) fall in anti-H. pylor IgG levels as a test of eradication in comparison to urease and histology. It was found that sensitivity was highest using a 10% cut-off but specificity wasvery low. Increasing cut-off values increased specificity but resulted in declining sensitivity without altering accuracy much. It appears that in the short term, percentage decline in serological titres can at best serve as a crude test of eradication.

Problem of timely diagnosis in anthrax meningitis.

Anthrax continues to remain a problem in parts of India. Meningitis is often a complication encountered among cases with cutaneous anthrax. We have encountered a dozen cases of anthrax meningitis in our hosptal in the past decade. A sudden unexplained rise in cases in the past two years with hundred percent mortality stresses the need for rapid confirmatory diagnosis. Most of the cases admitted with central nervous system involvement had a provisional diagnosis of conditions other than anthrax meningitis. A strong clinical suspicion with a simple Gram stain smear of the CSF will help confirm anthrax meningitis in endemic areas.

Inducible nitric oxide synthase gene and diabetic retinopathy in Asian Indian patients.

Nitric oxide, a signal transduction molecule, when modulated causes various diseases including diabetic retinopathy. In diabetes, allelic polymorphism of the inducible nitric oxide synthase (iNOS) gene is associated with retinopathy in the Northern Irish population. In the present study we investigated the Asian Indian population. One hundred and ninety-nine unrelated Asian Indian patients with 15 or more years of type 2 diabetes were divided into two groups: (a) diabetic retinopathy (DR) and (b) diabetic nonretinopathy (DNR) subjects. In these groups the pentanucleotide microsatellite repeat located 2.5 kb upstream of the transcription start site of the iNOS gene was amplified by polymerase chain reaction and analyzed. Eleven alleles, 175-225 bp, were identified. Allele 210 bp was significantly associated with retinopathy (p = 0.044). Individuals carrying this allele had twice the risk of developing retinopathy compared with those who did not carry this allele [odds ratio (OR) - 2.03; 95% CI 0.96-4.35]. Alleles 200 and 220 bp were also significantly associated with no retinopathy and no serious retinopathy complications, respectively. In the Asian Indian population, allele 210 bp of the iNOS gene is a high-risk allele for developing retinopathy and alleles 200 and 220 bp protect an individual from developing retinopathy or its complications.

Pneumolysin in urine: a rapid antigen detection method to diagnose pneumococcal pneumonia in children.

PURPOSE: Etiological diagnosis of pneumococcal pneumonia is difficult in small children in whom blood culture cannot be done or who have already been started on antibiotics. A simple technique which can be applied at the bedside or in the outpatient department may help in obviating this problem. Detection of pneumolysin, a product of invasive pneumococci is being exploited as a diagnostic tool. METHODS: An attempt was made to detect this protein in urine of seventy children, clinically suspected and radiologically diagnosed cases of pneumonia. Seventy age and sex matched controls were included in the study. Purified pneumolysin was prepared from clinical isolates of invasive pneumococcal infections. This was used to raise polyclonal antisera in rabbits. The antisera was used to sensitise Cowan 1 Staphylococcus aureus (CoA). A slide agglutination was performed with 25 microL urine and equal quantity of the reagent. RESULTS: Results were compared with CoA reagent sensitised with antisera raised against a genetically derived pneumolysoid and capsular polysaccharide for antigen detection in the urine. Pneumolysin could be detected in 42.9% (30/70) urine samples from cases with pneumonia by the genetically derived antigen and in 37.1% samples by the in house prepared antigen, in contrast to 2.1% in healthy controls and 4.2% in children with infections other than pneumonia. The result was statistically significant. Detection of pneumolysin was slightly better than detection of capsular polysaccharide antigen in urine although the result was not statistically significant. Blood culture proved to be positive in only 29.5% cases. CONCLUSIONS: Pneumolysin detection in urine showed promising results and was found to be simple and rapid. It will help in quickening the diagnosis of pneumococcal pneumonia.

Tumor necrosis factor allelic polymorphism with diabetic retinopathy in India.

The association of tumor necrosis factor (TNF) with diabetic retinopathy (DR) has been described previously. A total of 207 Asian Indian patients of 15-year duration of type 2 diabetes were identified. This group included (i) 100 patients with DR and (ii) 107 patients without retinopathy (DNR). In this study, we correlated the length of the (GT)n microsatellite di-nucleotide repeat upstream to the promoter region of TNF gene with susceptibility for the development of retinopathy. The microsatellite was polymerase chain reaction amplified and electrophoresed on polyacrylamide gel and silver stained. In our study population, there were 18 alleles ranging from 97 to 131 base pairs (bp). Allele 4 (103 bp) had a higher prevalence (9.81%) in the DNR group compared to that (2.5%) in the DR group (P=0.002). Patients with retinopathy and allele 8 (111 bp) had a tendency to develop proliferative diabetic retinopathy (PDR). In this study of Indian subjects, it is suggested that allele 4 is a low risk allele for developing retinopathy and allele 8 (111 bp) shows an association with PDR.