Biomedical subjects
S K Sethi
Publications and source records attributed to S K Sethi.
Vibrio cholerae diarrhoea in a three-day-old breastfed neonate.
A confirmed case of cholera in a 3-day-old neonate is being reported. Possible source of infection could be by holy water (Chamamrit) given to the baby, which is common ritual in India. Mother's milk has never been reported to transmit cholera, even though mother herself may be suffering from cholera. Contaminated water can transmit the disease as there is no maternally transmitted immunity.
Compound heterozygous familial hypercholesterolemia and familial defective apolipoprotein B-100 produce exaggerated hypercholesterolemia.
BACKGROUND: Familial hypercholesterolemia (FH) and familial defective apolipoprotein B-100 (FDB) represent ligand-receptor disorders that are complementary. Individuals with both FH and FDB are unusual. We report a family with both disorders and the impact of the mutations on the phenotypes of the family members. METHODS: We used single strand conformation polymorphism (SSCP) and denaturing gradient gel electrophoresis (DGGE) for genetic analysis of all 18 exons and the promoter region of the LDL receptor and DGGE for genetic analysis of the apolipoprotein B-100 (apo B-100) gene. The functional significance of the apo B-100 mutation was studied using a U937 cell proliferation assay. Fasting serum lipid profiles were determined for the index case and seven first-degree relatives. RESULTS: One of the patient's sisters had a missense mutation (Asp(407)-->Lys) in exon 9 of the LDL receptor and a serum LDL-cholesterol concentration of 4.07 mmol/L. Four other first-degree relatives had hyperlipidemia but no LDL-receptor mutation. However, these subjects had a mutation of the apo B-100 gene (Arg(3500)-->Trp). The cell proliferation rate of U937 cells fed with LDL from other subjects with the same mutation was fourfold less than that of controls. The index case had both FH- and FDB-related mutations. Her serum LDL-cholesterol (9.47 mmol/L) was higher than all other relatives tested. CONCLUSIONS: Existence of both FH and FDB should be considered in families with LDL-receptor mutations in some but not all individuals with hypercholesterolemia or when some individuals in families with FH exhibit exaggerated hypercholesterolemia.
Rhinocerebral mucormycosis in a patient of acute renal failure.
Mucormycosis is the name for invasive fungal infection caused by mucorales. The disease is uncommon and produces serious and rapidly fatal infection in patients with serious pre-existing illness. The classical presentation of rhinocerebral mucormycosis is involvement of nasal mucosa with invasion of paranasal sinuses and orbit. We report a case of mucormycosis in an otherwise healthy female who had developed acute renal failure following gastroenteritis.
Expression of intercellular adhesion molecule-1 (ICAM-1) in nasal epithelial cells of atopic subjects: a mechanism for increased rhinovirus infection?
Since clinical experimental studies indicate that upper respiratory tract viral infections may exacerbate acute asthma symptoms in atopic/asthmatic individuals, we have investigated the expression and modulation of ICAM-1 on human nasal epithelial cells (HNEC) from normal and atopic subjects. ICAM-1 is the attachment molecule for the majority of serotypes of human rhinovirus (HRV), including HRV-14, and is also critical for the migration and activation of immune effector cells. Basal ICAM-1 expression was significantly higher in HNEC obtained by brushings from atopic compared with non-atopic subjects (P = 0.031), and was also significantly increased on atopic HNEC harvested in season compared with out of season (P < 0.05). Atopic HNEC showed further up-regulation in ICAM-1 expression when cultured with clinically relevant allergen (P = 0.032). ICAM-1 levels on normal HNEC were also increased by infection with HRV-14 (P < 0.05). Basal expression of ICAM-1 on atopic nasal polyp epithelial cells (EC) was significantly higher than on both normal and atopic nasal HNEC. This elevated nasal polyp ICAM-1 level was not increased further by allergen, although HRV infection resulted in a small significant increase. Recovered viral titres from HRV-infected nasal polyp EC were 1.5-fold higher than from infected normal nasal HNEC. The data are consistent with the hypothesis that allergen, by enhancing expression of the HRV attachment target on host cells, facilitates viral infection in atopic subjects; simultaneously HRV-induced increases in ICAM-1 levels would favour migration and activation of immune effector cells to the airway, resulting in enhanced atopic inflammation.
Effects of intragenic variability at 3 polymorphic sites of the apolipoprotein B gene on serum lipids and lipoproteins in a multiethnic Asian population.
We determined the allelic (X+/X-, M+/M-, and E+/E-) distribution frequencies of the XbaI, MspI, and EcoRI restriction fragment length polymorphisms (RFLPs) in the apolipoprotein B gene in a control group of 374 healthy Chinese, Malays, and Indians and in a hyperlipidemic cohort of 131 Chinese patients. Covariability between the RFLPs and serum lipid, lipoprotein, and apolipoprotein concentrations was also studied. We found a lower frequency (average 0.0829) of the X+ allele and higher frequencies of the E+ (average 0.9452) and M+ (average 0.9772) alleles in our study population compared with frequencies reported in other populations. The 3 polymorphic sites did not contribute to significant variations in lipid levels (p > 0.1 in all cases). Also, there was no significant variation in genotype frequencies between the control subjects and the hyperlipidemic subjects. Despite their relative close proximity within the APOB gene sequence, the 3 polymorphic sites did not show any significant linkage disequilibrium. However, the presence of the X+ cutting site was in linkage disequilibrium with the Del allele of the 5' insertion-deletion polymorphism and the E-allele was in linkage disequilibrium with the 3' VNTR located near the 3' end of the coding region of the APOB gene.
Acute cerebellar ataxia following herpes zoster ophthalmicus.
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Th2 cytokines exert a dominant influence on epithelial cell expression of the major group human rhinovirus receptor, ICAM-1.
Intercellular adhesion molecule (ICAM)-1 is a cell receptor important in both human rhinovirus (HRV) attachment and immune effector cell mobilization. The level of expression of ICAM-1 by epithelial cells (EC) therefore plays a crucial role in the intricate biological phenomena underlying viral binding, host infection and consequent inflammatory events. As T-helper (Th)2 lymphocytes predominate within the asthmatic airway, the influence was evaluated of Th2-associated mediators in the modulation of ICAM-1 expression on uninfected and HRV-infected EC. H292 EC were cultured in vitro, with varying concentrations of interleukin (IL)-4, IL-5, IL-10 and IL-13 for 24 h and then infected with live HRV-14. Surface ICAM-1 expression was assessed by immunocytochemistry. Infection with HRV-14 resulted in a twofold increase in ICAM-1 expression. IL-4, IL-5, IL-10 and IL-13 produced a 2.7-5.1-fold enhancement of ICAM-1 expression of uninfected cells and caused approximately a further twofold increase in infected cells over the expression induced by HRV infection itself. Interferon-gamma in combination with each Th2-associated cytokine only slightly reduced, but did not override, the Th2-induced level of ICAM-1 expression on both uninfected and virus-infected EC. These data suggest that the effects of Th2-associated cytokines on intercellular adhesion molecule-1 expression and recovery of infectious virus are dominant over the effects of the Th1-associated cytokines such as interferon-gamma. Since the airway mucosa in atopic asthma is predominantly infiltrated by Th2 lymphocytes, these results could explain both the increased susceptibility to human rhinovirus infection in asthmatic patients and the associated exacerbation of asthma symptoms.
The StuI polymorphism on exon 8 of the low density lipoprotein (LDL) receptor gene: prevalence and impact on serum lipid levels in an Asian cohort.
A single nucleotide (A-->G) substitution in codon 370 (exon 8) of the low density lipoprotein (LDL) receptor gene results in loss of a StuI restriction site and an amino acid change (Ala370-->Thr) in the translated protein. This biallelic polymorphism has been associated with significant variations in plasma lipid concentrations in several Caucasian populations. We investigated its prevalence and impact on lipid metabolism in 539 Singaporeans of Chinese, Malay and Indian descent. The average frequency of 0.003 for the Thr370 allele and heterozygosity of 0.0056 were 10- to 20-fold lower than those reported in Caucasians (range 0.075 to 0.107). Distribution of the genotypes satisfied the Hardy-Weinberg equilibrium (chi 2 = 0.004, P = 0.948). We also found ethnic variation in the allele frequencies of Thr370 among the three races studied (0.009 in Indians, 0.007 in Malays, and 0.003 in Chinese). However, this needs to be confirmed with larger numbers. No significant correlation between genotype and serum concentrations of total cholesterol, triglyceride, high density lipoprotein (HDL)-cholesterol, LDL-cholesterol, lipoprotein(a), apolipoprotein A and apolipoprotein B was found in our study. This argues strongly against a major contribution or impact of this intragenic polymorphic locus on modulating the serum lipid profiles of the study subjects.
Interferon-gamma (IFN-gamma) down-regulates the rhinovirus-induced expression of intercellular adhesion molecule-1 (ICAM-1) on human airway epithelial cells.
Human rhinoviruses (HRV) are a major cause of upper respiratory tract infections in man, and can exacerbate existing pulmonary disease. The major group of HRV attach to ICAM-1, which is expressed on nasal and bronchial epithelial cells. To study the influence of biological mediators on ICAM-1 expression, and consequently HRV attachment and infection, we compared the effects of various cytokines, alone and in combination, on ICAM-1 expression by an uninfected and HRV-infected bronchial epithelial cell line H292. Cytokines known to be released soon after viral infection, such as tumour necrosis factor-alpha (TNF-alpha), IL-1beta and the chemokine IL-8 increase ICAM-1 expression on uninfected cells. Epithelial cells infected with live HRV-14 displayed marked up-regulation of ICAM-1 compared with baseline. TNF-alpha further enhanced the HRV-induced increase in ICAM-1 expression on epithelial cells, peaking at day 4 after infection, whilst IL-8 exhibited a steady increase in ICAM-1 expression over 14 days. In contrast, IFN-gamma, a known Th1 antiviral lymphokine, whilst increasing the level of ICAM-1 on uninfected cells, induced a significant persistent down-regulation of ICAM-1 expression on HRV-infected epithelial cells. With combinations of TNF-alpha and IFN-gamma, ICAM-1 expression on HRV-infected cells was reduced to basal levels. The effects of IFN-gamma were paralleled by a reduction in viral titres. Our in vitro model has provided useful insights into the early pathogenic events of HRV infection at the level of the host cell-virus interaction. Our data confirm that biological mediators play a crucial role in the pathogenesis as well as the course of HRV infection which is modulated by the types, and time kinetics of inflammatory cytokines in the immediate microenvironment.
Denaturing gradient-gel electrophoresis screening of familial defective apolipoprotein B-100 in a mixed Asian cohort: two cases of arginine3500-->tryptophan mutation associated with a unique haplotype.
The Arg-to-Trp substitution at codon 3500 in the apolipoprotein (apo) B-100 gene is established as a cause of familial defective apo B-100 (FDB), a functional mutation, resulting in reduced LDL receptor binding and manifest hypercholesterolemia. In a search for similar mutations in 163 Malaysians, we screened the putative receptor-binding region (codons 3456-3553) of the apo B-100 gene by PCR amplification and denaturing gradient-gel electrophoresis. Four single-base mutations were detected and confirmed by DNA sequencing. Two females, a Chinese and a Malay, had the same CGG3500-->TGG mutation, resulting in an Arg3500-to-Trp substitution. This is the second published report of such an independent mutation involving the same codon as the established Arg3500-to-Gln mutation. The two other mutations detected, CTT3517-->CTG and GCC3527-->GCT, resulted in degenerate codons with no amino acid substitutions. All four mutations were associated with a unique apo B haplotype, different from those found in Caucasian FDB patients but concurring with that previously reported for two other Asians with FDB.
Analysis of polycyclic aromatic hydrocarbons in metered dose inhaler drug formulations by isotope dilution gas chromatography/mass spectrometry.
Organic compounds extracted into metered dose inhalers (MDIs) from the rubber components of the metering valve are of increasing interest in the development of these formulations. Polycyclic aromatic hydrocarbons (PAHs) are a class of extractable organic compounds whose source is the carbon black commonly used as a reinforcing agent in rubber. The analytical method for PAHs described in this report employs "cold filtration" to remove the suspended drug substance and excipients, and gas chromatography/mass spectrometry (GC/MS) for separation and detection of individual PAHs. After filtration, stable isotope labelled analogues of target PAHs are spiked into the drug product to act as internal standards, correcting for recovery (termed "isotope dilution GC/MS"). Validation of the method was accomplished with respect to linearity, precision, limit of detection/quantitation, selectivity and ruggedness. Application to a variety of MDI drug product formulations revealed that certain PAHs are present at the ng/inhaler level.
Use of an alkaline phosphatase-labeled synthetic oligonucleotide probe for detection of Campylobacter jejuni and Campylobacter coli.
A commercially available synthetic nucleic acid probe (SNAP) conjugated to alkaline phosphatase was compared with standard culture techniques for detecting Campylobacter species. The SNAP was able to detect either 5 ng of C. jejuni DNA or 10(5) CFU of bacteria. The SNAP could also detect DNA extracted from 10(5) CFU in mock-infected stool samples. The SNAP detected C. jejuni and C. coli but showed no reactivity with C. laridis, C. fetus subsp. fetus, C. fetus subsp. venerealis, C. fennelliae, "C. upsaliensis," C. cinaedii, C. fecalis, C. hyointestinalis, C. mucosalis, or Helicobacter (Campylobacter) pylori. The SNAP also showed no cross-reactivity with other enteric pathogens. When applied to pure cultures, the SNAP detected 55 clinical isolates of C. jejuni and 11 clinical isolates of C. coli, with an accuracy of 100%. When applied directly to clinical specimens, the SNAP detected Campylobacter spp. in 19 of 23 culture-positive stool specimens (sensitivity, 82.6%; specificity, 100%). Pure cultures of the Campylobacter strains isolated from the four probe-negative, culture-positive stool specimens gave positive reactions with the SNAP. While the SNAP had excellent sensitivity and specificity for isolated bacterial colony isolates, the main limitation to the Campylobacter probe detection kit may be the sensitivity limit on direct detection of Campylobacter organisms in stools.
Retinal hemorrhages after extra corporeal membranous oxygenation.
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Outbreak of neonatal listeriosis in a regional hospital in Kuwait.
Six newborns with Listeria monocytogenes infection were admitted to the same neonatal unit between 31 October and 3 December 1985. The index case, a preterm baby, was born to a mother who was febrile with an influenza-like illness at the time of delivery. This baby presented with Listeria sepsis and pneumonia. Another child was born from whose mother L. monocytogenes was isolated from the cervix with the same serotype as that in the CSF of her newborn. In the other cases blood and cervical cultures of the mothers were negative, while Listeria was isolated from the CSF of their babies. Five out of six infants developed meningitis between 9 and 12 days after birth. All isolates were serotype 4b and indistinguishable by phage typing. All babies were successfully treated without any major sequelae. Although cross-infection was strongly suggested, the source of the outbreak could not be established. This is the first documented report of human listeriosis in Kuwait.
Microbial etiology of acute gastroenteritis in hospitalized children in Kuwait.
During a period of 15 months 621 hospitalized children with acute gastroenteritis and 152 control children were investigated for etiologic agents of the disease. Putative enteropathogens were identified in 86% of the patients and 10% of the controls. Common viral agents associated with gastroenteritis among children included rotaviruses (45%) and enteric adenoviruses (4%). Bacterial pathogens infecting children were Salmonella serotypes (24%), enterotoxigenic Escherichia coli (9%), Campylobacter jejuni (7%), enteropathogenic E. coli (7%), Shigella (4%) and enterotoxigenic Aeromonas sp. (1%). The highest incidence of infections was observed in the 3- to 25-month age group. Mixed infections were observed in 12% of the patients. Viral gastroenteritis was clinically mild and of short duration. Upper respiratory tract infections, vomiting and watery stools were common features. In contrast bacterial gastroenteritis was more severe; stools were frequently bloody and abdominal pain, cramps, shock, convulsions and milk intolerance were predominant clinical features. Comparative analysis revealed differential features of bacterial and viral gastroenteritis which should help clinicians to make a tentative diagnosis and to start treatment early.
Detection of human rotavirus by using an alkaline phosphatase-conjugated synthetic DNA probe in comparison with enzyme-linked immunoassay and polyacrylamide gel analysis.
An alkaline phosphatase-conjugated synthetic oligodeoxyribonucleotide probe was compared with polyacrylamide gel electrophoresis (PAGE) detection of rotavirus RNA as well as an enzyme-linked immunosorbent assay (ELISA) for the detection of rotavirus in stools from young children with gastroenteritis. The synthetic probe did not cross-react with bacterial causative agents of diarrheal disease. Extraction of viral RNA from stool samples with a phenol-chloroform mixture was suitable for most samples. In some cases fecal pigments interfered with the reaction of the probe with viral RNA. The use of ion-exchange chromatography to further purify viral RNA removed contaminating pigments and increased the sensitivity of the probe assay. Of 260 stool specimens, 77 (30%) were positive for rotavirus when tested by PAGE analysis of rotavirus RNA. The synthetic probe identified 71 rotavirus specimens when RNA obtained by phenol-chloroform extraction followed by chromatographic purification was used (sensitivity, 91.0%; specificity, 96.7%). The ELISA results also agreed well with the electrophoretic analysis (sensitivity, 98.7%; specificity 94%) and the probe assay (sensitivity, 90%; specificity, 100%). Discordant results between the ELISA and the probe assay were examined further by electron microscopy and PAGE analysis of viral RNA. The positive and negative predictive values of the probe assay in comparison with PAGE were 92.2 and 96.1%, respectively. Rotaviruses showing both long and short RNA electrophoretic patterns were detected by the probe. The probe assay coupled with chromatographic purification of rotavirus RNA is an effective method for detecting rotavirus and compares favorably with PAGE analysis and ELISA.
Molecular epidemiology of human rotavirus infections based on genome segment variations in viral strains.
Molecular epidemiology of rotavirus infections in 621 hospitalized children was investigated by analysis of migration patterns of viral genomic ribonucleic acid (RNA) segments by electrophoresis in polyacrylamide gels. Based on migration patterns of RNA segments of 184 rotavirus strains, seven different electropherotypes were identified: 146 (79.3%) strains were "long," and 38 (20.7%) were "short" electropherotypes; 61% belonged to a single dominant "long" electropherotype, which persisted throughout the 15-month period of study, whereas six other cocirculating types appeared at varying intervals. Electrophoretic migration patterns of RNA from viral isolates of two patients suggested mixed infections with different rotaviruses. There was a lack of correlation between the electrophoretic migration of segments 10 and 11 and serologically defined subgroup specificity in three of the rotavirus strains. Rotavirus infections and different electropherotypes were observed throughout the year.