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Biomedical subjects

A Ayyagari

Publications and source records attributed to A Ayyagari.

At least 19 recordsLinked to original sources

In vitro (1)H NMR studies of RD human cell infection with echovirus 11.

The effects of echovirus 11 infection on RD human cell line (derived from rhabdomyosarcoma) were studied using (1)H NMR spectroscopy and optical microscopy. Both uninfected and infected cells consumed glucose and produced lactate, acetate and formate as extracellular metabolites. In infected whole cells, phosphocholine and uridine-sugar were observed in addition to the metabolites observed in uninfected cells. Water-soluble intracellular metabolites of infected cells showed glutamine, phosphocholine and glycine which were not observed in uninfected cells. Cellular metabolites except lipid components gradually decreased and disappeared during 24-48 h of viral infection. The quantity of lipid components in infected cells was comparable with that in uninfected cells, indicating that echovirus 11 does not utilize cell lipid molecules. Unlike optical microscopy, (1)H NMR spectroscopy identified early stages of infection through metabolic changes. These results may have potential implications in probing virus-cell interactions using NMR-based metabolomics.

Cell Line, Tumor↗

Nucleotide polymorphism associated with ethambutol resistance in clinical isolates of Mycobacterium tuberculosis.

Ethambutol (EMB) is a first-line drug used for antitubercular therapy in combination with other drugs as recommended by World Health Organization DOTS/DOTS-Plus regimens. EMB is also effective in the treatment of opportunistic mycobacterial infections in patients with human immunodeficiency virus. The emb locus has been considered as a drug target for EMB, and substitutions of codon 306 in Mycobacterium tuberculosis gene embB have been shown to be the most frequent and predictive mutations for EMB resistance. The aim of the present study was to detect embB and embC gene mutations in EMB-resistant clinical isolates. A total of 23 isolates of M. tuberculosis from patients with pulmonary tuberculosis were included in the study. Drug sensitivity was tested by proportion method and E-test. All 23 isolates were EMB resistant. Primers to amplify the embB and embC gene were designed, and polymerase chain reaction products were subjected for sequence analysis. H37Rv standard laboratory strain was used as control. Nucleotide sequencing showed that 16 strains had a mutation in the embB gene. The most common mutation observed in the embB gene was at codon 306, followed by mutations at codons 299 and 378 in 4 and 2 isolates, respectively. Novel mutations have been reported at codons 239, 240, 247, 282, 311, 368, 397, 446, 469, and 471. Sequence analysis of the embC gene showed mutation in 8 isolates at codon 270. Novel mutations in embC have been reported at codons 251 and 254. The most common nucleotide polymorphism in our isolates was at codons 306 and 299 in the embB gene and at codon 270 in the embC gene. A mutation at codon 306 was usually associated with high-level ethambutol resistance.

Antitubercular Agents↗

Antibiotic susceptibility of Helicobacter pylori clinical isolates: comparative evaluation of disk-diffusion and E-test methods.

Antimicrobial susceptibility of 25 Helicobacter pylori strains isolated from patients with acid peptic diseases were tested for in vitro sensitivity to commonly used antibiotics using disk-diffusion and E-test, methods. All strains tested were susceptible to tetracycline by E-test, with the minimum inhibitory concentration (MIC) values being <0.125 microg/ml for all strains except for 6 (<0.023 microg/ml). However 1 strain was resistant by disk-diffusion method. One strain was resistant to clarithromycin both by disk diffusion and E-test (MIC <48 microg/ml), and 1 strain was resistant only by disk diffusion. Only one strain was resistant to amoxicillin by disk diffusion and E-test (MIC >256 microg/ml). For ciprofloxacin, three strains were resistant by disk diffusion and two by E-test (MIC <32 microg/ml). Sixteen strains were resistant to metronidazole by disk diffusion and E-test (MIC >or= 8 microg/ml), and 1 was resistant only by E-test (MIC <48 microg/ml). Overall, 64% of the strains were resistant to metronidazole. The MIC for metronidazole was also tested by agar-dilution method, and metronidazole resistant strains had an, MIC >8 microg/ml. The disk-diffusion method showed excellent correlation with E-test results; there was 100% agreement for amoxicillin a other antibiotics showed 90% to 95% accuracy. Disk diffusion is cheaper than E-test (approximately 2.6 cents vs. 2.60 US dollars), is easy to perform, and is a reliable method for testing H. pylori susceptibility to antimicrobial agents in the clinical microbiology laboratory.

Anti-Bacterial Agents↗

Development of a simple restriction fragment length polymorphism assay for subtyping of coxsackie B viruses.

Coxsackie B viruses (genus, Enterovirus; family, Picornaviridae) can cause aseptic meningitis, encephalitis, pleurodynia, myocarditis and are implicated in the pathogenesis of dilated cardiomyopathy. The differentiation of the group B coxsackieviruses into their subtypes has potential clinical and epidemiological implications. In the present study, a simple restriction fragment length polymorphism (RFLP) assay was developed for typing of group B coxsackieviruses into subtypes 1-6. It is a two step process, first, virus isolation and identification by virus neutralization assay, using pools of polyclonal antisera, second, the reverse transcription polymerase chain reaction (RT-PCR) using a single primer pair selected from the conserved 5'-untranslated region (5'-UTR) of enterovirus genome followed by RFLP. A 440 bp product was amplified from the reference strains of each subtype of group B coxsackievirus and 29 clinical isolates (positive for group B coxsackieviruses by neutralization assay). The amplified products were subjected to restriction endonuclease digestion by enzyme BsaJI. The assay was able to distinguish all six serotypes of coxsackie B viruses. The results were comparable to serotyping and showed that due to the relatively conserved nature of 5'-UTR in enterovirus genome, this region can be used for subgeneric molecular identification of enteroviruses.

5' Untranslated Regions↗

Fungal peritonitis in patients on continuous ambulatory peritoneal dialysis: a single centre Indian experience.

BACKGROUND: Fungal peritonitis (FP) is a serious complication in patients on continuous ambulatory peritoneal dialysis (CAPD). We reviewed our FP cases to analyse the causative agents and possible risk factors in relation to FP and its outcome and mortality. METHODS: Records of all FP cases were reviewed. FP was diagnosed based on effluent cell count and positive fungal culture in suitable media. RESULTS: Between October 1993 and November 2001, 261 patients underwent CAPD. FP was detected in 28 patients, one episode in each patient (14.3% of the total peritonitis episodes). Candida species and dematiaceous fungi+/-Candida species were responsible for 89.3 and 10.7% of episodes, respectively. Patients with preceding bacterial peritonitis (BP) developed FP more frequently (25.6%) than de novo cases (2.9%) (P<0.0001) and lower proportion of them continued CAPD (8.6% vs. 60%; P=0.007). Mortality in patients having abdominal pain with and without fever, and catheter in situ was significantly higher than in those patients who did not have these risk factors (9/11 vs. 6/17, P=0.01; 13/17 vs. 2/11, P=0.003; 6/6, vs. 9/22, P=0.01, respectively). CONCLUSIONS: Higher proportion of our patients had FP; preceding BP was a significant risk factor for development of FP and technique failure. Abdominal pain+/-fever in patients and catheter in situ were identified as risk factors associated with mortality.

Female↗

Molecular characterization of echovirus 11 isolates from India.

BACKGROUND & OBJECTIVES: Echovirus 11 (ECV11) is one of the most frequent non-polio enteroviruses isolated from stool samples of children with acute flaccid paralysis in north India. The present work was undertaken to study the sequence variability in the 440 bp of 5'-non-translated region of ECV11 genome using heteroduplex mobility assay (HMA). METHODS: Twelve ECV11 isolates were studied for sequence variability in the 5'-non-translated region (5'NTR) using the HMA followed by nucleotide sequencing. HMA was used to determine sequence diversity between Indian ECV11 isolates and prototype Gregory strain. HMA results were confirmed by 5'NTR nucleotide sequencing of five Indian ECV11 isolates. RESULTS: HMA results showed high genomic diversity between the prototype Gregory strain and Indian ECV11 isolates. All isolates were grouped into five different types of heteroduplex mobility patterns with respect to Gregory strain. A 440 bp 5'NTR fragment of five ECV11 isolates representing different heteroduplex patterns, was sequenced. The sequence alignment showed that 5'NTR of Indian isolates was different from prototype Gregory strain and identical to the ECV11 isolates of Finland and Hungary. Phylogenetic analysis including ECV11 isolate sequences from different parts of the world showed that Indian ECV11 isolates represented a different subgroup. INTERPRETATION & CONCLUSION: The results of the present study suggested that the HMA could be successfully used as a preliminary screening method for sequence variability determination of enterovirus field isolates. The sequence data generated on ECV11 isolates from India will be useful for future studies of endemic genotypes of echovirus.

5' Untranslated Regions↗

Use of telemedicine in evading cholera outbreak in Mahakumbh Mela, Prayag, UP, India: an encouraging experience.

Telemedicine (TM) services a process in which expert medical advice from afar is provided using electronic signals to transfer the medical data from one site to another. As a pilot project to assess the efficacy of TM in developing countries like India, a telemedicine center was set up at the main hospital of Mahakumbh mela--a grand religious fair, at Prayag, a city in north India. The daily reporting of the in-patient and outpatient cases at the fair revealed a surge of diarrhea cases among the pilgrims at the fair. This information was communicated to the referral center at Sanjay Gandhi Post Graduate Institute of Medical Sciences (SGPGIMS), which, with the help of its microbiology department, conducted microbiological examinations of stool samples and rectal swabs of patients along with various water samples. Vibrio cholerae was isolated in 22.6% (7/31) of the samples. This information was immediately relayed to the Main Hospital at the fair online, and then to the health authorities, who took strict and prompt measures to improve hygiene. Subsequently, the number of diarrhea cases decreased considerably in a matter of a few days, and thus an epidemic disaster was averted, which could have created havoc in such a large gathering.

Aged↗

Candidaemia in patients of a tertiary health care hospital from north India.

BACKGROUND & OBJECTIVES: Candidaemia is an important cause of mortality in hospital settings. Limited information is available from India on nosocomial candidaemia. The objective of the present study was to isolate and identify yeasts from patients suspected to have nosocomial bloodstream infection (BSI) and to determine the carriage rate of Candida species, risk factors for acquisition of infection and mortality in this group of patients. METHODS: Blood samples from 4871 patients suspected to have BSI at least 48 h after admission were cultured following standard protocol to isolate and identify the pathogens. Clinical details, possible risk factors and outcome of all candidaemic patients were recorded and analysed. Samples of hand washings and throat gargles from these patients were also cultured to determine the carriage rate. Candida albicans isolated from patients and their carriage sites were genotyped by randomly amplified polymorphic DNA (RAPD) analysis to study strain relatedness. RESULTS: Twenty one patients with candidaemia were detected with mortality of 55 per cent. Candidaemia per 1000 admissions was 1.61. Isolation of non-C. albicans Candida species was significantly higher than C. albicans (14/21 vs 7/21: P < 0.05). Use of broad-spectrum antibiotics (43%), gastrointestinal surgery (23%), immunosuppressive therapy (23%), protein calorie malnutrition with parenteral hyperalimentation (23%) and neutropaenia (14%) were identified as probable risk factors. The seven C. albicans strains isolated from patients with BSI were typed into 6 genotypes. Yeast carriage rate among the patients was 71.4 per cent. C. albicans isolated from the hand, throat and blood of two patients had identical genotype. INTERPRETATION & CONCLUSION: BSI due to non-C. albicans Candida species is more common than C. albicans in our patients and candidaemia is associated with high mortality. RAPD appears to be a simple method to study strain relatedness for C. albicans. There is a need for early diagnosis and systematic surveillance to meet the challenges of nosocomial candidaemia.

Adolescent↗

Cryptococcal infection in patients with clinically diagnosed meningitis in a tertiary care center.

A retrospective analysis of 326 clinically diagnosed cases with meningitis over a period of five-and-a-half years was carried out to determine the prevalence of cryptococcal infection, its associated risk factors and therapeutic outcome. Fifty-four (16.6%) patients with cryptococcal meningitis were identified by smear examination, culture and/or cryptococcal antigen latex agglutination test. Records of 45 cryptococcal meningitis patients were available; 18 (40%) of them were apparently healthy immunocompetent individuals, 13 (28.9%) had human immunodeficiency virus (HIV) infection, 9 (20%) were renal transplant recipients, 4 (8.9%) were diabetic and 1 (2.2%) had systemic lupus erythematosus. Ten (22.2%) patients died and 11 (24.4%) patients (all HIV-positive) left against medical advice. The present study indicates that cryptococcal infection is associated with high mortality. Presenting symptoms being indistinguishable from other causes of central nervous system infection, all patients with a clinical diagnosis of meningitis, irrespective of their immune status should be investigated for cryptococcal infection.

Adolescent↗

Transmission of filarial infection through blood transfusion.

Microfilariae can be transmitted by blood transfusion and they may be circulated in the recipient's blood but they do not develop into adult worms. Mortality associated with transfusion associated filarial infection is not documented but it may give rise to morbidity in transfusion recipients in terms of allergic reaction. The present study was carried out to investigate the association of post transfusion reactions and filarial infections in an endemic area. About 11,752 transfusion recipients were followed up and in 15 months period, 47 (0.4%) post transfusion reactions (PTR) were reported. Routine investigations for post transfusion reaction were carried out in all 47 patients and their respective blood donor. Moreover, blood culture, microfilaria detection by concentration technique, filarial antibody and antigen detection (both by ELISA) were done in all subjects. Out of 47 patients showing post transfusion reaction, 29 (61.7%) patients developed allergic reaction. Eighteen (38.3%) patients having allergic reaction did not have previous history of blood transfusion and 14 (29.8%) of them received transfusion from blood donors who was either positive for microfilaria, filarial antigen or antibody. Microfilaremia was demonstrated in 4 (8.5%) patients and 5 (10.6%) blood donors. Microfilaria was concurrently present in 2 patients and their respective donors. Filarial antibody was detected in 27 (56.5%) patients and 26 (55.3%) blood donors but microfilaria was detected in 3 (6.4%) and 4 (8.5%) subjects, respectively. Antigen detection test correlated with microfileraemic state of subjects. The result shows that transfusion associated filarial infection may be a probable cause for transfusion-associated morbidity in endemic areas. In 14 (29.8%) patients having allergic reactions, the probable cause was transfusion-associated filarial infection. Filarial antigen detection test was found to be more useful in detecting infections. Blood donors with active history of filarial infection should be deferred from donating blood. Filarial antigen detection test may be employed as screening test for blood donors, if possible.

Animals↗

UreC PCR based diagnosis of Helicobacter pylori infection and detection of cag A gene in gastric biopsies.

Polymerase chain reaction assay using ureC gene specific primers for the detection of Helicobacter pylori in gastric biopsy specimens from 116 dyspeptic patients was compared with other routine invasive diagnostic methods (culture, rapid urease test [RUT] and histology). In parallel, gastric biospy specimens from 54 patients and their corresponding Helicobacter pylori isolates were subjected to PCR with cagA targeting primers using standard protocols. Helicobacter pylori were detected in 53%, 43%, 48% and 50% of patients by PCR, RUT, culture and histological examination respectively. Based on histology and culture positive and at least three test positive result, 44 (37%), 46 (39%) and 26 (22%), and 56 (48%), 52 (44%) and 8 (6%) patients were classified as Helicobacter pylori positive, negative and indeterminate respectively. The sensitivity and specificity of PCR assay was the highest-95% and 100% when compared with both culture and histology positive, and at least any three positive results respectively. The result of cagA positivity in 54 gastric biopsy specimens and their corresponding Helicobacter pylori isolates were identical; 18 of 20 (90%) duodenal ulcer patients and 23 of 28 (82%) patients with chronic gastritis and 2 (40%) of 5 patients with portal hypertension and one gastric biopsy specimens from gastric cancer patients were found to be cagA positive. PCR-based method to detect Helicobacter pylori and the virulence gene cag A directly from gastric biopsy specimens appears to be promising and can curtail the lengthy process of culture-based approaches. The procedure proved to be rapid and reliable and could be utilized for diagnostic purposes.

Adult↗

Non-polio enteroviruses in acute flaccid paralysis.

OBJECTIVE: Human enteroviruses are the major cause of aseptic meningitis and also cause a wide range of other acute illnesses, including neonatal sepsis like disease, meningitis, acute flaccid paralysis and acute hemorrhagic conjunctivitis. Infection in neonates is particularly life threatening. METHODS: Stool samples of 523 children (age < 4 years) showing symptoms of acute flaccid paralysis (AFP) were studied. National Polio Surveillance Project workers from different parts of Uttar Pradesh and Bihar collected the samples during June to October 1998. Non-polio enteroviruses (NPEV) were isolated in 191 cases only, by cell culture based neutralization assay. These NPEV isolates were further studied to find the frequent enterovirus serotype detected in stool of children having AFP. RESULTS: Data generated will help future studies on NPEV serotypes circulating in this area. CONCLUSION: In addition it may reduce unnecessary hospitalization, allow immune globulin batches of high titres to frequently circulating serotypes, to be reserved for intravenous therapy of neonates and guide the formulation of antigens for rapid and less expensive diagnosis.

Child, Preschool↗

Isolated Pneumocystis carinii infection of adrenal glands causing Addison's disease in a non-immunocompromised adult.

Pneumocystis carinii is primarily an opportunistic pathogen infecting patients with AIDS and other immunocompromised patients, and ordinarily does not affect immunocompetent persons. We report isolated P. carinii infection of bilateral adrenal glands in a non-immunocompromised adult male, leading to fatal Addisonian crisis. Diagnosis of P. carinii was established on the basis of cytopathology and microbiological tests, using conventional staining techniques and direct immunofluorescence on ultrasound-guided fine needle aspirates and trucut needle biopsy specimen from adrenal glands. P. carinii pneumonia and other fungal infections of the adrenal glands were excluded by appropriate tests. Absence of HIV infection was established by negative ELISA for HIV I and II antibodies and Western blot analysis at the time of presentation and 45 d later. Normal blood total leukocyte and CD4 lymphocyte counts and IgG and IgA levels confirmed the immunocompetent status of the patient. The patient improved with anti-Pneumocystis treatment and corticosteroid replacement, but succumbed to an episode of Addisonian crisis triggered by a diarrheal illness.

Addison Disease↗

Campylobacter species associated with diarrhoea in patients from a tertiary care centre of north India.

BACKGROUND & OBJECTIVES: Most laboratories do not routinely distinguish the various Campylobacter species, though almost all Campylobacter species have been isolated from human faeces. The epidemiological and clinical aspects of its infection and the species involved in genesis of diarrhoea are least understood in the developing countries. The aim of the present study was to find out frequency of Campylobacter species isolated from patients with diarrhoea over a 12-year period and to analyse their features. METHODS: Campylobacter strains isolated from stool samples of patients with diarrhoea were identified to the species level on appropriate media at 42 degrees C micro-aerobically. Patients' demography and clinical data were analyzed retrospectively; 25 Campylobacter jejuni strains were tested for toxin production and 23 strains were typed by Penner scheme. RESULTS: A total of 62 strains were isolated from 59 patients and the various species were C. jejuni 51 (82.3%), C. coli 8 (12.9%), C. lari 2 (3.2%), and C. upsaliensis 1 (1.6%). Children < 5 yr of age were most affected (34/59; 57.6%), followed by patients in 15-30 yr of age (12/59; 20.3%). Presentation of watery diarrhoea was significantly more common than inflammatory diarrhoea (50/59, 84.7% vs 9/59, 15.3%; P < 0.001). Recurrence occurred in 3 (5.1%) patients. Majority of the infections resolved within one week; one HIV-positive patient had chronic diarrhoea. Two patients developed Guillain-Barré syndrome following Campylobacter infection. Twenty (80%) of 25 strains were toxigenic and 20 (87%) of 23 strains could be typed by Penner scheme. INTERPRETATION & CONCLUSION: In our patients, 4 different Campylobacter species and various C. jejuni serotypes were involved in gastroenteritis. Majority of the infections were watery diarrhoea and in children < 5 yr of age. There is a need of a population-based systematic study to know the epidemiology of whole spectrum of campylobacters in India.

Academic Medical Centers↗

Identification of enteric pathogens in HIV-positive patients with diarrhoea in northern India.

Enteric pathogens associated with chronic diarrhoea in HIV-positive patients were studied. The study was conducted during January 1995-December 1998. Stool specimens from all diarrhoea patients (n = 26) were examined microscopically for ova and parasites using wet preparations and stained smears. Stool samples from diarrhoea patients were also cultured on appropriate media to isolate enteric bacterial pathogens. Of the 59 patients, 26 (44%) had prolonged diarrhoea for more than 4 weeks. Enteric pathogens were detected in 19 (73%) of the 26 patients: 17 patients harboured a single pathogen, and 2 patients had mixed pathogens. The detection rate of emerging parasites, including Isospora, Cryptosporidium, Blastocystis hominis, and Strongyloides stercoralis as a single agent, was significantly higher than conventional pathogens (50% vs 19.2%; p < 0.05). Only one patient harboured both conventional and emerging pathogens (Entamoeba histolytica and Cryptosporidium). Isospora belli was detected in 8 (31%) of the 26 diarrhoea patients: in 7 (27%) patients as a single agent and in one patient with S. stercoralis. Cryptosporidium was identified in 3 (11%) diarrhoea patients: in 2 (8%) patients as a single agent and in one patient with E. histolytica, followed by B. hominis in 2 (8%) patients. E. histolytica was most commonly isolated (3/26; 11.5%), followed by Giardia lamblia, enteropathogenic Escherichia coli, and Campylobacter jejuni (one patient each). Parasitic pathogens were frequently associated with HIV-positive patients with diarrhoea in northern India. I. belli was the most frequent parasite isolated, followed by Cryptosporidium. Stools of all HIV-positive patients with diarrhoea should thoroughly be investigated to identify aetiologic agents for proper management.

AIDS-Related Opportunistic Infections↗

Comparison of various conventional methods with a polymerase chain reaction assay for detecting methicillin-resistant & susceptible Staphylococcus aureus strains.

BACKGROUND & OBJECTIVES: Methicillin-resistant Staphylococcus aureus (MRSA), a major nosocomial pathogen world-wide, is often difficult to detect due to the heterogeneous nature of expression of oxacillin resistance. In the present study, various conventional methods were compared with polymerase chain reaction on 106 clinical isolates of Staph. aureus for detection of oxacillin resistance. METHODS: A total of 106 clinical isolates of Staph. aureus were tested for oxacillin resistance by disc diffusion, screen agar plates (3 micrograms and 6 micrograms/ml of oxacillin), oxacillin broth (3 micrograms/ml) and mecA based PCR. RESULTS: PCR detected mecA gene amplified product of 604 bp in 57 strains. Disc diffusion failed to detect 7 mecA positive strains but identified 5 mecA negative strains as oxacillin resistant. Screen agar 3 micrograms, screen agar 6 micrograms and oxacillin broth 3 micrograms detected 55, 53 and 55 respectively of the 57 mecA positive strains; however, they also falsely identified 5, 3 and 3 strains of mecA negative strains respectively as oxacillin resistant. The sensitivity, specificity and accuracy of disc diffusion, 3 micrograms screen agar, 6 micrograms screen agar and 3 micrograms oxacillin broth against PCR as gold standard were as follows: 87.7, 89.9 and 88.7 per cent; 96.5, 89.8 and 93.4 per cent; 93.0, 93.9 and 93.4 per cent; 96.5, 93.9 and 95.3 per cent respectively. INTERPRETATION & CONCLUSIONS: The present study demonstrated that disc diffusion test was least reliable and 3 micrograms broth had the highest sensitivity and specificity when compared with PCR for detection of oxacillin resistance. Because of variations among the methods, a combination of tests should be used for the accurate detection of MRSA till new guidelines by an appropriate body are formulated.

Base Sequence↗

Primary subcutaneous Nocardia asteroides infection in a renal allograft recipient.

Nocardia asteroides is an important opportunistic pathogen in immunocompromised hosts. The primary infection is usually in the lungs and is followed by dissemination to other parts of the body. Primary subcutaneous infection with Nocardia asteroides has been reported rarely (three reports) and no such case has been reported in a renal transplant recipient. We describe here a case of renal transplant recipient who developed primary subcutaneous infection with Nocardia asteroides within one and half years of the transplantation.

Dermatomycoses↗