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Biomedical subjects

P Datta

Publications and source records attributed to P Datta.

At least 55 records · Page 3Linked to original sources
PubMed1997

Nasopharyngeal pneumococcal colonization among Kenyan children: antibiotic resistance, strain types and associations with human immunodeficiency virus type 1 infection.

OBJECTIVES: To compare pneumococcal nasopharyngeal colonization rates among HIV-1-infected children with those of uninfected children born to seropositive mothers and those of seronegative controls. To determine the predominant serotypes and antimicrobial susceptibility among pneumococcal isolates in Kenya. METHODS: Nasopharyngeal pneumococcal colonization was examined in 207 children recruited from the Perinatal HIV-1 Transmission Study conducted in Nairobi, Kenya. Colonization was compared among HIV-1-infected children, uninfected children born to seropositive mothers and control seronegative children. Isolates were serotyped and tested for antibiotic susceptibility to penicillin, tetracycline, erythromycin, chloramphenicol, clindamycin and rifampin. RESULTS: Colonization was higher among HIV-1-infected and uninfected children than among controls only when associated with respiratory illnesses (86% of 7 and 60% of 20 vs. 29% of 31, P = 0.004). No differences were observed when children were asymptomatic (20% of 35, 35% of 94 and 22% of 101). Intermediate penicillin resistance was found in 60% of 94 isolates, 28% were resistant to tetracycline and all isolates were susceptible to the other antibiotics tested. Sixteen serotypes were identified, with 13, 15, 14, 6B and 19F comprising 73% of isolates. Serotype 13 was found in 31% of colonized children. This serotype and 2 others isolated are not found in the current 23-valent polysaccharide vaccine. Overall 41% of colonized children harbored nonvaccine strains. CONCLUSIONS: Although nasopharyngeal pneumococcal colonization was high among children with respiratory illness born to HIV-1-seropositive mothers, increased asymptomatic colonization did not explain the increased risk of invasive pneumococcal disease associated with HIV-1 infection. Intermediate penicillin resistance was common but high level penicillin and multiple antibiotic resistance were not seen. The prevalence of the unique strains circulating in this region will need to be considered in the design of effective pneumococcal vaccines for use in East Africa.

Acquired Immunodeficiency Syndrome↗
PubMed1997

Analytical performance of a new chemiluminescent phenytoin (ACS:180) assay.

The authors, as a beta testing site, evaluated the ACS:180-phenytoin chemiluminescent assay (Ciba-Corning Diagnostics Corp., Medfield, MA, U.S.A.) by comparing its performance with a widely used fluorescence assay for phenytoin (Abbott Laboratories, Abbott Park, IL, U.S.A.). The ACS:180-phenytoin assays were run on a ACS-180 analyzer and fluorescence polarization assays on a TDx analyzer. The within-run precision for ACS-phenytoin assay was determined using controls obtained from Ciba-Corning. The CVs were 2.9% for low control (mean = 5.5, SD = 0.16 microgram/ml, n = 10), 2.8% for the medium control (mean = 13.4, SD = 0.37 microgram/ml, n = 10), and 2.7% for the high control (mean = 24.6, SD = 0.66 microgram/ml, n = 10). The corresponding between run precisions were 4.1% for the low control (mean = 5.4, SD = 0.22 mg/ml, n = 10), 3.1% for the medium control (mean = 13.8, SD = 0.43 mg/ml, n = 10), and 2.9% (mean = 24.5, SD = 0.70 mg/ml, n = 10) for the high control. The assay was linear from 0.5 to 40 micrograms/ml of serum phenytoin concentrations with a detection limit of 0.24 microgram/ml. The recoveries were 93-97% for concentrations of phenytoin of 5-30 micrograms/ml. They also compared 111 serum specimens collected from patients receiving phenytoin. The concentrations of phenytoin ranged from none detected to 32.4 micrograms/ml. Using fluorescence polarization assay as x-axis (reference method) and ACS:180-phenytoin assay as y-axis, they obtained the following regression line: y = 1.0x - 0.26, r = 0.993. They conclude that the ACS-phenytoin assay has a good precision and that the results correlate well with the fluorescence polarization assay.

Fluorescence Polarization↗
PubMed1997

Interference of oleandrin and oleandrigenin in digitoxin immunoassays: minimal cross reactivity with a new monoclonal chemiluminescent assay and high cross reactivity with the fluorescence polarization assay.

Toxicity from ingestion of the oleander plant is common. Oleandrin, the oleander glycoside, has structural similarity to cardiac glycoside digoxin and is known to cross react with various digoxin immunoassays. The authors studied the cross reactivity of oleandrin and its deglycosylated congener oleandrigenin with a fluorescence polarization immunoassay for digitoxin and compared their results with a new chemiluminescent assay for digitoxin on the Automated Chemiluminescent System (ACS:180 Plus) from Chiron Diagnostics. Even though the chemiluminescent assay has been reported to be comparable with the fluorescence polarization assay among normal patient population, oleandrin and oleandrigenin showed very high cross reactivities with the fluorescence polarization immunoassay and minimal cross reactivity with the new chemiluminescent assay. When the authors supplemented a serum specimen containing no digitoxin with 50 micrograms/ml of oleandrin, the fluorescence polarization assay recorded a value of 535.7 ng/ml of digitoxin equivalent, whereas the new chemiluminescent assay recorded a value of 10.3 ng/ml of digitoxin equivalent. The cross reactivity of oleandrigenin with the fluorescence polarization immunoassay for digitoxin was significantly lower than oleandrin. The presence of oleandrin also falsely elevated total digitoxin level in a specimen supplemented with digitoxin and oleandrin. The authors also measured free digitoxin concentration by the fluorescence polarization immunoassay in the ultrafiltrate of serum supplemented with digitoxin and oleandrin. Because digitoxin and oleandrin are bound strongly to protein, monitoring free digitoxin concentration by the fluorescence polarization immunoassay instead of total digitoxin concentration does not eliminate oleandrin interference. The authors conclude that fluorescence polarization immunoassay for digitoxin has a high cross reactivity with oleandrin and can falsely elevate digitoxin concentration in the presence of oleandrin, whereas the new chemiluminescent assay for digitoxin is almost free from interferences from oleandrin.

Anti-Arrhythmia Agents↗
PubMed1997

Involvement of Fnr and ArcA in anaerobic expression of the tdc operon of Escherichia coli.

Anaerobic expression of the tdcABC operon in Escherichia coli, as measured by LacZ activity from single-copy tdc-lacZ transcriptional and translational fusions, is greatly reduced in strains lacking two global transcriptional regulators, Fnr and ArcA. The nucleotide sequence of the tdc promoter around -145 shows significant similarity with the consensus Fnr-binding site; however, extensive base substitutions within this region had no effect on Fnr regulation of the tdc genes. A genetic analysis revealed that the effect of Fnr on tdc is not mediated via ArcA. Furthermore, addition of cyclic AMP to the anaerobic incubation medium completely restored tdc expression in fnr and arcA mutants as well as in strains harboring mutations in the Fnr- and ArcA-dependent pfl gene and the Fnr-regulated glpA and frd genes. These results, taken together with the earlier finding that tdc expression is subject to catabolite repression by intermediary metabolites, strongly suggest that the negative regulatory effects of mutations in the fnr and arcA genes are mediated physiologically due to accumulation of a metabolite(s) which prevents tdc transcription in vivo.

Amino Acid Transport Systems, Neutral↗
PubMed1996

Comparison of four digoxin immunoassays with respect to interference from digoxin-like immunoreactive factors.

OBJECTIVES: Comparison of a new monoclonal digoxin assay with three polyclonal digoxin assays for their cross-reactivity to digoxin-like immunoreactive factors (DLIF) and digoxin metabolites. DESIGN AND METHODS: Sixty-six nondigitalized patient samples from 5 different groups: neonates, women in 3rd trimester pregnancy, and patients with liver or renal diseases, or undergoing organ transplants, and 139 samples from digoxin-treated patients of 4 categories (hospital sick, liver, renal, and outpatients) were compared in 4 different digoxin assays: (a) ACS Digoxin (ACS) developed for the automated chemiluminescent Ciba Corning ACS 180 system, (b) Baxter Stratus (Stratus, a fluoroimmunoassay), (c) Ciba-Corning Magic (Magic, a radioimmunoassay), and (d) an in-house radioimmunoassay (RIA). The ACS and RIA were also compared for their cross-reactivity to four principal digoxin metabolites. RESULTS AND CONCLUSION: Among the nondigitalized specimens, no significant DLIF interference was found for all 4 assays among the pregnant women or liver and transplant patients. However, the neonates registered high DLIF interference with Magic and RIA, but none for ACS or Stratus. DLIF interference in renal samples was highest in the Magic assay and lowest in RIA. Among the specimens with digoxin, a higher number of discrepant samples were found from the sick patients than from outpatients. In 75% of such discrepant samples, the ACS result was less than other assay results, suggesting DLIF as the probable cause. The two assays differed most in their cross-reactivity to the deglycated metabolites, digoxigenin and its mono-digitoxoside.

Adult↗
PubMed1996

Estimating concentrations of total digoxin and digoxin-like immunoreactive substances in volume-expanded patients being treated with digoxin.

High concentrations of digoxin-like immunoreactive substances (DLIS) artificially increase serum digoxin concentrations. However, DLIS are absent in the protein-free ultrafiltrate because of their strong binding with serum macromolecules, whereas approximately 75% of digoxin can be found in the ultrafiltrate. Using regression analysis, we devised equations by which total digoxin concentration can be calculated from free digoxin and albumin concentrations in serum. We used two different assays, fluorescence polarization and chemiluminescence, for measuring total and free-digoxin concentrations in sera. Both equations were very similar. Because measured concentrations of digoxin in the serum exhibit DLIS interferences, the measured concentrations were sometimes higher in volume-expanded patients than the calculated digoxin concentrations. We also estimated the extent of interferences from DLIS by subtracting the calculated digoxin concentration from the measured digoxin concentration in volume-expanded patients.

Cardenolides↗
PubMed1996

Effect of antibody specificity on results of selected digoxin immunoassays among various clinical groups.

We examined the specificity of three automated digoxin immunoassays (Abbott TDxFLx Digoxin II assay, Baxter-Dade Stratus II Digoxin assay, and Ciba Corning ACS Digoxin assay) applied without modification to (a) sera from 229 digoxin-free patients in 12 cohorts associated with nonspecific or endogenous digoxin-like immunoreactive factor (DLIF) interference, and (b) drug-free serum supplemented with the major metabolites and analogs of digoxin. We observed three patterns of apparent digoxin results among the DLIF samples: one common to kidney and liver failure patients, where TDx and Stratus assays showed significant positive results; one common to newborns and cord blood, where only the TDx assay had significant interference; and one from cardiac surgery patients, where the Stratus assay alone showed interference. Of the three assays, the ACS had the least interference from DLIF. The assays also behaved differently with respect to cross-reactivity with digoxin metabolites, digitoxin, and digitoxin metabolites. The ACS assay again had the least analog or metabolite cross-reactivity. The three methods agreed well on digoxin-positive specimens, with a mean bias of <0.15 microgram/L digoxin for each and discrepancies (defined as >3 SD between the assay pairs compared) of only 3-5%.

Antibody Specificity↗
PubMed1995

Influence of DNA topology on expression of the tdc operon in Escherichia coli K-12.

TdcB activity expressed from the chromosomal gene and LacZ expression from single-copy tdc-lacZ transcriptional and translational fusions were measured in Escherichia coli strains harboring mutations in the genes encoding DNA gyrase, topoisomerase I and the HU protein. The pattern of tdc operon expression in these mutants suggests that relaxation of supercoiled DNA enhances tdc transcription in vivo.

DNA, Bacterial↗
PubMed1994

Mother-to-child transmission of human immunodeficiency virus type 1: report from the Nairobi Study.

Mother-to-child transmission of human immunodeficiency virus type 1 (HIV-1) is a significant problem in countries with endemic HIV-1 infection. Between 1986 and 1991, 365 children of HIV-1-infected mothers and 363 control children were studied in Kenya. The overall risk of transmission from mother to child, determined by serologic evidence of infection by age > or = 12 months and excess mortality in the HIV-1-exposed group, was 42.8% (range, 27.6%-62.2%). Marriage was the only maternal characteristic associated with transmission (odds ratio, 2.2; 95% confidence interval, 1.2-4.2; P < .05). Children who experienced growth failure were more likely to be infected. In 44% of children ultimately infected, the pattern of antibody response implied intrapartum or postnatal exposure to HIV-1. Of potential postnatal exposures examined, duration of breast-feeding beyond age 15 months and the mother being married were independently associated with increased risk of infection and seroconversion of children. The percentage of HIV infection attributable to breast-feeding > or = 15 months was 32%. The frequency of mother-to-child transmission of HIV-1 was high; a substantial proportion of infection occurred postnatally, possibly through breast-feeding.

Acquired Immunodeficiency Syndrome↗
PubMed1994

Human immunodeficiency virus type 1 IgA antibody in breast milk and serum.

Breast-feeding plays a potentially significant role in mother to child transmission of human immunodeficiency virus type 1 (HIV-1). The additional transmission risk attributable to breast-feeding and the factors that enhance or inhibit transmission are presently unknown. One mechanism by which breast milk might inhibit HIV-1 transmission is the presence of specific antibodies directed against HIV-1 in breast milk of seropositive mothers. In this study serum and breast milk samples from women in Nairobi, Kenya, were tested to determine the prevalence of HIV-1 IgA antibodies. A Western blot test developed in our laboratory was used to detect anti-HIV-1 immunoglobulin A in serum and anti-HIV-1 secretory IgA (sIgA) in breast milk. Ninety-four percent of 63 HIV-1 seropositive women had anti-HIV-1 IgA in serum and 59% had anti-HIV-1 sIgA in their breast milk. No significant associations with maternal characteristics or serum anti-HIV-1 IgA or IgG banding patterns and the presence of anti-HIV-1 sIgA in breast milk were found. No protective effect of anti-HIV-1 sIgA was seen regarding mother to child transmission; however, further studies are necessary to determine the effect of these antibodies in maternal sera or in breast milk on the efficacy of HIV-1 transmission.

AIDS Serodiagnosis↗
PubMed1994

Functional analysis of the tdcABC promoter of Escherichia coli: roles of TdcA and TdcR.

The efficient expression of the tdc operon of Escherichia coli requires the products of two regulatory genes, tdcR and tdcA. We have identified the transcription site of tdcR by primer extension mapping and established the translation start site of TdcR by mutational analysis of its reading frame. In a tdcR tdcABC deletion strain, tdcR+ promoted high-level LacZ expression from a lambda tdcAB-lacZ lysogen and mutations introduced in tdcR resulted in a greater than sixfold decrease in LacZ level. In-frame deletions of tdcA also reduced LacZ expression, and chromosomal and plasmid-borne tdcA+ increased the LacZ level in tdcA mutant lysogens. Interestingly, multicopy tdcA+ plasmids introduced into tdcR mutant strains completely restored tdc expression. In separate experiments we found that mutations in the tdc promoter DNA around positions -70, -140, and -175 greatly reduced tdc expression relative to that for the wild-type promoter and the tdcP mutation around -175 prevented multicopy tdcA+ from rescuing tdcR mutants. Furthermore, competition experiments revealed that a wild-type promoter fragment encompassing the -175 region cloned into a plasmid reduced tdc expression by titrating TdcA in vivo, and this effect was reversed with excess TdcA. These results suggest that in tdcR+ cells TdcR interacts with tdcP and/or TdcA to enhance tdc transcription whereas in tdcR mutant cells a new tdcP-TdcA complex around -175 in the native promoter bypasses the requirement for TdcR. On the basis of the accumulated data summarized here and elsewhere we propose that multiple transcription factors enhance tdc operon expression by bending and looping of the promoter DNA to form an active transcription complex.

Bacterial Proteins↗
PubMed1993

TdcA, a transcriptional activator of the tdcABC operon of Escherichia coli, is a member of the LysR family of proteins.

The tdcB and tdcC genes of the tdcABC operon of Escherichia coli encode threonine dehydratase and a threonine-serine permease, respectively. These proteins are involved in transport and metabolism of threonine and serine during anaerobic growth. In this study, we functionally characterized tdcA, which encodes a 35 kDa polypeptide consisting of 312 amino acid residues. Non-polar and partially polar mutations introduced into tdcA drastically reduced the expression of the genes down-stream from tdcA. Complementation studies using single-copy chromosomal integrants of a tdcB-lacZ fusion harboring an in-frame deletion of tdcA with chromosomal or plasmid-borne tdcA+ in trans showed complete restoration of tdc operon expression in vivo. The amino acid sequence at the amino-terminal end of TdcA revealed a significant homology to the helix-turn-helix motifs of typical DNA binding proteins. Sequence alignment of TdcA with LysR also showed considerable sequence similarity throughout their entire lengths. Our results suggest that TdcA is related to the LysR family of proteins by common ancestry and, based on its functional role in tdc expression, belongs to the LysR family of transcriptional activators.

Amino Acid Sequence↗
PubMed1992

Increased risk of early measles in infants of human immunodeficiency virus type 1-seropositive mothers.

An increase in illness due to measles is one of the potential consequences of the human immunodeficiency virus (HIV) epidemic in Africa. During a study of perinatal HIV transmission conducted in Kenya, the risk of acquiring measles before vaccination (9 months of age) was found to be 3.8 times higher in infants born to HIV-seropositive mothers than in control infants (10 [9%] of 109 vs. 5 [3%] of 194 infants; P = .02; odds ratio, 3.8; 95% confidence interval, 1.2-13.2). The majority of infants who developed measles in this study had significant sequelae related to their measles infection. The increased risk of measles appeared to be related to relatively lower anti-measles antibody titers detected in cord blood samples of affected infants born to HIV-seropositive mothers. However, 94% of all infants were susceptible to measles on the basis of ELISA testing at age 6 months regardless of maternal HIV serology. These observations highlight the need for improved measles vaccination strategies in Africa and for studies to delineate the effects of HIV infection on the incidence, presentation, and sequelae of childhood infectious illnesses.

Antibodies, Viral↗
PubMed1992

Integration host factor is required for positive regulation of the tdc operon of Escherichia coli.

A 14-bp segment in the promoter region of the tdcABC operon of Escherichia coli shows sequence identity with the consensus binding site for the E. coli integration host factor (IHF). In an himA (IHF-deficient) strain, expression of beta-galactosidase from a tdcB'-'lacZ protein fusion plasmid was about 10% of that seen with an isogenic himA+ strain. Threonine dehydratase activity from the chromosomal tdcB gene in the himA mutant was also about 10% of the wild-type enzyme level. Two different mutations introduced into the putative IHF-binding site in the fusion plasmid greatly reduced the plasmid-coded beta-galactosidase activity in cells containing IHF. In vitro gel retardation and DNase I footprinting analyses showed binding of purified IHF to the wild-type but not to the mutant promoter. IHF protected a 31-bp region between -118 and -88 encompassing the conserved IHF consensus sequence. These results suggest that efficient expression of the tdc operon in vivo requires a functional IHF and an IHF-binding site in the tdc promoter.

Amino Acid Sequence↗
PubMed1992

Catabolite gene activator protein and integration host factor act in concert to regulate tdc operon expression in Escherichia coli.

Anaerobic expression of the tdcABC operon of Escherichia coli requires cyclic AMP and the catabolite gene activator protein (CAP). Purified CAP binds to a 30-bp sequence in the tdc promoter between positions -55 and -26, and a mutant CAP site with base substitutions at positions -48, -47, and -45 failed to bind CAP and also drastically reduced the beta-galactosidase expression from a tdcB'-'lacZ fusion plasmid. Recently, we showed that efficient expression of the tdc operon also requires a functional integration host factor (IHF) and an IHF-binding site in the tdc promoter between positions -118 and -88. The levels of beta-galactosidase activity from the tdcB'-'lacZ fusion plasmids were also reduced in an IHF-deficient strain with the wild-type or mutant plasmid CAP sequence. In vitro footprinting experiments revealed that CAP and IHF occupy their specific binding sites on tdc DNA when they are present separately or together. These regulatory proteins also induced significant bending of the tdc promoter DNA. Our results suggest that CAP and IHF act in concert as positive transcription factors for tdc operon expression in vivo.

Anaerobiosis↗