Enzyme immunoassay for detection of antibody-coated bacteria.

To quantitatively evaluate factors potentially affecting antibody coating of bacteria in urine, we developed an assay with enzyme-linked rather than fluorescein-conjugated immunoglobulin. Using the enzyme immunoassay (EIA) in an in vitro system in which concentrations of serotype O44 Escherichia coli and antibody titer to E. coli Orr O44 O antigen were known, we compared specimens run in parallel with a fluorescent antibody (FA) assay. At greater than or equal to 10(5) bacteria per ml, antibody titer to homologous O antigen correlated directly with absorbance in the EIA. Both tests had sensitivities exceeding 95% in specimens containing greater than or equal to 10(5) bacteria per ml, but the FA test detected 23 of 27 positive specimens with less than 10(5) bacteria per ml compared with 21 of 43 detected by EIA (P = 0.002). However, nonspecific fluorescence caused false positives in 8% of negative tests run by FA compared with 1% of simultaneous EIA tests (P = 0.05). pH alterations and pretreatment of bacteria with antibiotics did not affect either test. Heterologous E. coli strains showed no cross-reactivity with O44 antiserum, but all Staphylococcus aureus isolates tested caused false positives in both assays, and one Klebsiella strain repeatedly caused a false-positive FA assay. The EIA appears to be a simple, quantitative, and specific technique for detection of antibody-coated bacteria in this experimental system.     

Ligase chain reaction to detect Chlamydia trachomatis infection of the cervix.

We performed a multicenter evaluation of ligase chain reaction (LCR) in the diagnosis of Chlamydia trachomatis infection of the cervix. This LCR provides an amplification of target sequences within the chlamydial cryptic plasmid. The LCR results were compared with those of isolation in cell culture. Discrepant (tissue culture-negative and LCR-positive) test results were resolved by the application of a direct immunofluorescent-antibody test to detect chlamydial elementary bodies and by the use of alternate DNA primers that targeted the chlamydial major outer membrane protein gene. A total of 234 of 2,132 specimens (10.9%) could be confirmed as containing C. trachomatis. Of these, 152 were detected by isolation in cell culture and 221 were detected by LCR. The corresponding sensitivities were 94% for LCR and 65% for cell culture. There was greater variability among study site results for cell culture sensitivity (52 to 92%) than for LCR sensitivity (87 to 98%). The specificity of each test was greater than 99.9%. Thus, LCR offers a highly sensitive nonculture method for detecting chlamydial infection of the cervix.     

Association of carboxylesterase B electrophoretic pattern with presence and expression of urovirulence factor determinants and antimicrobial resistance among strains of Escherichia coli that cause urosepsis.

We determined the carboxylesterase B electrophoretic profiles of 74 blood isolates of Escherichia coli from patients with urosepsis. Most strains (64%) exhibited the B2 electrophoretic pattern. P fimbrial and hemolysin genetic determinants were present and expressed significantly more often among strains with the B2 than with the B1 electrophoretic pattern. In contrast, aerobactin determinants were significantly more prevalent and more commonly expressed among the B1 strains; this difference was attributable to the presence of plasmid-encoded aerobactin in one-third of the B1 strains (P = 0.02, B1 versus B2). The prevalence and extent of antimicrobial resistance was significantly greater among the B1 strains, and the B1 electrophoretic pattern was more often found in isolates from patients with urinary tract abnormalities. We conclude that the carboxylesterase B electrophoretic pattern differentiates two groups of E. coli isolates from patients with urosepsis: strains with the B1 electrophoretic pattern are associated with urologically impaired hosts, characteristically lack P fimbrial and hemolysin determinants, and often carry a plasmid-encoded aerobactin system (possibly on multiple antimicrobial resistance plasmids), whereas B2 strains more commonly invade noncompromised hosts, express P fimbriae and hemolysin, carry chromosomal aerobactin determinants, and lack antimicrobial resistance.     

Use of a urine enzyme immunoassay as a diagnostic tool for Chlamydia trachomatis urethritis in men.

We collected first-voided urine specimens from 659 males attending a sexually transmitted disease clinic and performed both enzyme immunoassay (EIA) for detection of chlamydial antigen and leukocyte esterase testing on these urine samples. The overall prevalence of chlamydial urethritis in the study population as determined by culture of urethral swabs was 11%. However, 46% of all men in the study had no symptoms of urethritis. Compared with urethral cultures for chlamydiae, the urine EIA had a sensitivity of 42% and a specificity of 99%. The sensitivity of the EIA strongly correlated with the amount of antigen present in culture as assessed by numbers of inclusion-forming units. The sensitivity of the leukocyte esterase test compared with that of chlamydia culture was 88%. We conclude that in this population of men, which included many patients without symptoms of urethritis, the urine EIA was a relatively insensitive means of screening for chlamydial infection.     

Characterization of recast Nafion films by small- and wide-angle X-ray scattering

Room temperature recast Nafion

1 General formula:

films, prepared under different humidities in the curing atmosphere, were characterized by small- and wide-angle X-ray scattering (SAXS and WAXS). SAXS reveals that the humidity influences the size of the ionic clusters in these polymer films. It was found that the number and size of ionic clusters increases with increasing relative humidity. WAXS indicates the presence of locally ordered regions of the backbone polymer. These results allow a qualitative understanding of the electrochemical behaviour of Nafion recast films cured at different humidities, as described in the literature.     

Association of total and central adiposity measures with fasting insulin in a biracial population of young adults with normal glucose tolerance: the CARDIA study.

OBJECTIVE: To determine the association of computed tomography (CT)-measured visceral adipose tissue (AT) and other measures of adiposity with fasting insulin in a biracial (African American and Caucasian) study population of young adults. RESEARCH METHODS AND PROCEDURES: The study population consisted of 251 young adults with normal glucose tolerance (NGT), ages 28-40 years, who were volunteers from the Birmingham, Alabama, and Oakland, California centers of the Coronary Artery Risk Development in Young Adults (CARDIA) study. RESULTS: In regression models with total adiposity measures (body mass index or dual-energy X-ray absorptiometry-measured percent fat), visceral AT (measured as a cross-sectional area in cm2) was generally a stronger predictor of insulin than overall adiposity in all race/gender groups (partial correlation coefficients ranging from 0.31 to 0.47) except for black men, in whom the associations were nonsignificant. Partial correlation coefficients between waist circumference and insulin, controlling for percent fat, were nearly identical to those between visceral AT and insulin in women and in white men. Analyses performed on 2060 NGT CARDIA subjects who were not in this study of visceral AT showed significant correlations of waist circumference with insulin in all race/gender groups, including black men, and that black men in the visceral AT study group were significantly leaner than other black male CARDIA subjects. DISCUSSION: We conclude that visceral AT was associated with fasting insulin in NGT participants in three of the four race/gender groups (black men excepted) and that waist circumference was a good surrogate for visceral AT in examining associations of central adiposity with fasting insulin.     

Coronary artery bypass procedures in septuagenarians are justified. Short and long-term results

BACKGROUND: Elective coronary artery bypass surgery can be performed with low operative mortality. There is a controversial discussion whether short-term and long-term results of CABG justify this procedure in elderly patients. METHODS: To add to this discussion, we evaluated retrospectively, the clinical profile, operative technique, short- and long-term results of 1127 patients over 70 years of age who underwent myocardial revascularization between January 1985 and December 1996. RESULTS: Mean age of the cohort was 73.9 years and it rose continuously. In 1996, septuagenarians represented 21.5% of our coronary patients, in comparison to 6.4% in 1985. Analysis of risk factors showed an increasing prevalence of renal failure, obesity, hyperuricemia and a history of smoking. Preoperatively, 87% of our patients were in NYHA-class III or IV, and thus operated to relieve severe symptoms. The percentage of emergent operations decreased over the observed period by 10.3%. The internal mammary artery was used with rising frequency (44.8% in 1985-1992 vs 61.5% in 1993-1996). The number of simultaneous valve replacements increased, too. Postoperatively, we noted a rising incidence of respiratory failure (17.1%) and neurological disorders (13.7%). On the other hand, the need for intra-aortic balloon pumping and hemofiltration declined by 6.6% and 2.9%, respectively. Perioperative mortality (< or =24 hrs) was 3.65%, hospital mortality (< or =30 days) was 9.64%. The actuarial survival (standard error) at 1 year was 82% (+/-4.3%), and 65.7% (+/-3.8%) at 5 years. CONCLUSIONS: Our data suggest that coronary artery bypass grafting can be performed in septuagenarians at an acceptable risk. Since the large majority of patients improve symptomatically, surgery is a recommendable option for a growing number of elderly patients suffering from severe angina.     

Outcomes of a randomized trial of hyperfractionated cranial radiation therapy for treatment of high-risk acute lymphoblastic leukemia: therapeutic efficacy and neurotoxicity.

PURPOSE: We evaluated 8-year survival and late neuropsychologic toxicity in children with acute lymphoblastic leukemia treated in a randomized clinical trial to test whether hyperfractionated (twice daily) cranial radiation therapy (CRT) can reduce incidence and severity of late toxicities associated with 18 Gy of CRT. PATIENTS AND METHODS: Between 1987 and 1995, 369 children treated on two consecutive Dana-Farber Cancer Institute Consortium protocols for high-risk acute lymphoblastic leukemia were randomly assigned to conventionally fractionated CRT (CFX) or hyperfractionated CRT (HFX) to a total dose of 18 Gy. Neuropsychologic testing was completed for 125 of 287 children in continuous complete remission. Event-free and overall survival, as well as neuropsychologic function, were compared for the two arms of the protocol. RESULTS: Eight-year event-free survival (+/- SE) was 80% +/- 3% for children randomly assigned to CFX and 72% +/- 3% for HFX (P =.06). Overall survival was 85% +/- 3% for CFX and 78% +/- 3% for HFX (P =.06). CNS relapses occurred in 2.8% of patients receiving CFX and 2.7% receiving HFX (P =.99). Cognitive function for both groups was solidly in the average range, with no group differences in intelligence, academic achievement, visuospatial reasoning, or verbal learning. Children on the HFX arm exhibited a modest advantage for visual memory (P <.05). CONCLUSION: HFX provides no benefit in terms of cognitive late effects and may compromise antileukemic efficacy. HFX should not be substituted for conventionally dosed CRT in children who require radiation therapy for treatment of acute lymphoblastic leukemia.     

Comparison of ciprofloxacin (7 days) and trimethoprim-sulfamethoxazole (14 days) for acute uncomplicated pyelonephritis pyelonephritis in women: a randomized trial

Context  The optimal antimicrobial regimen and treatment duration for acute uncomplicated pyelonephritis are unknown. Objective  To compare the efficacy and safety of a 7-day ciprofloxacin regimen and a 14-day trimethoprim-sulfamethoxazole regimen for the treatment of acute pyelonephritis in women. Design  Randomized, double-blind comparative trial conducted from October 1994 through January 1997. Setting  Twenty-five outpatient centers in the United States. Patients  Of 378 enrolled premenopausal women aged at least 18 years with clinical diagnosis of acute uncomplicated pyelonephritis, 255 were included in the analysis. Other individuals were excluded for no baseline causative organism, inadequate receipt of study drug, loss to follow-up, no appropriate cultures, and other reasons. Interventions  Patients were randomized to oral ciprofloxacin, 500 mg twice per day for 7 days (with or without an initial 400-mg intravenous dose) followed by placebo for 7 days (n = 128 included in analysis) vs trimethoprim-sulfamethoxazole, 160/800 mg twice per day for 14 days (with or without intravenous ceftriaxone, 1 g) (n = 127 included in the analysis). Main Outcome Measure  Continued bacteriologic and clinical cure, such that alternative antimicrobial drugs were not required, among evaluable patients through the 4- to 11-day posttherapy visit, compared by treatment group. Results  At 4 to 11 days posttherapy, bacteriologic cure rates were 99% (112 of 113) for the ciprofloxacin regimen and 89% (90 of 101) for the trimethoprim-sulfamethoxazole regimen (95% confidence interval [CI] for difference, 0.04-0.16; P = .004). Clinical cure rates were 96% (109 of 113) for the ciprofloxacin regimen and 83% (92 of 111) for the trimethoprim-sulfamethoxazole regimen (95% CI, 0.06-0.22; P = .002). Escherichia coli, which caused more than 90% of infections, was more frequently resistant to trimethoprim-sulfamethoxazole (18%) than to ciprofloxacin (0%; P<.001). Among trimethoprim-sulfamethoxazole–treated patients, drug resistance was associated with greater bacteriologic and clinical failure rates (P<.001 for both). Drug-related adverse events occurred in 24% of 191 ciprofloxacin-treated patients and in 33% of 187 trimethoprim-sulfamethoxazole–treated patients, respectively (95% CI, -0.001 to 0.2). Conclusions  In our study of outpatient treatment of acute uncomplicated pyelonephritis in women, a 7-day ciprofloxacin regimen was associated with greater bacteriologic and clinical cure rates than a 14-day trimethoprim-sulfamethoxazole regimen, especially in patients infected with trimethoprim-sulfamethoxazole–resistant strains.