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Antibiotic susceptibility testing of Mycoplasma genitalium by TaqMan 5' nuclease real-time PCR.

Mycoplasma genitalium is an important pathogen in male nongonococcal urethritis (NGU). Isolation of M. genitalium from clinical specimens by axenic culture is very difficult and time-consuming, and very few strains are available for antibiotic susceptibility testing. Primary isolation of M. genitalium by coculture with Vero cells improves the isolation rate significantly. However, some strains cannot be adapted to axenic culture. In this study, we determined the antibiotic susceptibility of M. genitalium strains grown in Vero cell culture with dilutions of antibiotics. Growth of M. genitalium was monitored by a quantitative PCR assay detecting a single-copy region of the mgpB adhesin gene. Growth inhibition in the presence of antibiotics was expressed as a percentage of the DNA load of controls grown in the absence of antibiotics. Eighteen strains were examined, including 6 new strains isolated from urethral swab specimens and 4 new strains isolated from urine specimens collected from Japanese men. Eight strains adapted to axenic culture were also tested by the conventional broth dilution method. The two methods had an acceptable correlation. Azithromycin was the most active drug against M. genitalium. Among the fluoroquinolones, moxifloxacin had the highest activity, with MICs ranging from 0.03 to 0.5 mg/liter, whereas ciprofloxacin and levofloxacin were considerably less active, with MICs ranging from 0.5 to 16 mg/liter and 0.25 to 4 mg/liter, respectively. MICs for tetracycline ranged from 0.125 to 4 mg/liter. This new method could increase the number of M. genitalium strains available for antibiotic susceptibility testing and significantly shorten the time from sampling to MIC results.

Animals↗

Parent-to-child transmission is relatively common in the spread of the human polyomavirus JC virus.

JC polyomavirus (JCV), the causative agent of progressive multifocal leukoencephalopathy, is ubiquitous in the human population, infecting children asymptomatically and then persisting in renal tissue. In most adults, renal JCV replicates and the progeny are excreted in urine. We used this urinary JCV to elucidate the routes of JCV transmission. A 610-bp JCV DNA region (IG region) encompassing the 3'-terminal sequences of both T-antigen and VP1 (major capsid protein) genes was amplified by means of PCR from urine specimens collected from all members of seven families. JCV strains were then unequivocally identified by the nucleotide sequences of the amplified IG regions. We could identify 18 distinctive JCV strains from 27 individuals. Different JCV strains were detected from all unrelated persons. However, the same viral strain was detected from one (four families), two (one family), or three offspring (one family) as well as from the fathers (three families) or from the mothers (three families). In total, the JCV strains detected in half of the JCV-positive children were identified in their parents. Since most humans are infected during childhood, these findings indicated that JCV is transmitted frequently from parents to children. We roughly estimated that 50% of JCV transmission occurs by this route and that the other 50% occurs outside the family.

Adult↗

Bacterial susceptibility to oral antibiotics in community acquired urinary tract infection.

BACKGROUND: The most common oral antibiotics used in the treatment of urinary tract infection (UTI) are sulphonamides and cephalosporins, but emerging resistance is not unusual. AIMS: To assess the change in susceptibility of urinary pathogens to oral antibiotics during the past decade in children with community acquired UTI. METHODS: The study sample included two groups of children with a first community acquired UTI: 142 children enrolled in 1991 and 124 enrolled in 1999. UTI was diagnosed by properly collected urine specimen (suprapubic aspiration, transurethral catheterisation, or midstream specimen in circumcised males) in symptomatic patients. Antimicrobial susceptibility of the isolates was compared between the two groups. RESULTS: The pathogens recovered in the two groups were similar: in 1991--E coli 86%, Klebsiella 6%, others 8%; in 1999--E coli 82%, Klebsiella 13%, and others 5%. A slight but generalised decrease in bacterial susceptibility to common antibiotics in the two groups was shown: ampicillin 35% versus 30%; cephalexin 82% versus 63% (p < 0.001); nitrofurantoin 93% versus 92%. The only exception was co-trimoxazole, 60% versus 69%. Overall resistance to antibiotics in 1999 was as follows: ampicillin 70%, cephalexin 37%, co-trimoxazole 31%, amoxicillin-clavulanate 24%, nitrofurantoin 8%, cefuroxime-axetil 5%, nalidixic acid 3%. CONCLUSIONS: This study shows a slight but generalised decrease in bacterial susceptibility to common oral antibiotics in the past decade in our population. Empirical initial treatment with co-trimoxazole or cephalexin is inadequate in approximately one third of UTI cases. A larger number of pathogens may be empirically treated with amoxicillin-clavulanate (24% resistance); 95% of organisms are susceptible to cefuroxime-axetil.

Administration, Oral↗

Pregnancy diagnosis in the common marmoset (Callithrix jacchus jacchus).

Abdominal palpation of the uterus was carried out on 25 pregnant and 29 non-pregnant marmosets (Callithrix jacchus jacchus). 195 complete 24-hour urine specimens, collected between the first week of gestation and term (21 weeks), were tested with the Sub-Human Primate Tube (SHPT) test. No significant differences between the two methods in their ability to diagnose pregnancy were obtained between the first and 13th week of gestation. The SHPT test was seldom positive after the 13th week of pregnancy, whereas, with one exception, palpation always indicated pregnancy.

Animals↗

Bence Jones proteins and light chains of immunoglobulins. XV. Effect of corticosteroids on synthesis and excretion of Bence Jones protein.

The effect of corticosteroids and cytotoxic chemotherapeutic agents on the excretion of Bence Jones protein was determined for periods of 1 - 62 mo in 29 patients with multiple myeloma and Bence Jones proteinuria. The amount of protein present in 24-h urine specimens collected before treatment and at frequent intervals during monthly treatment cycles was determined. Striking variations occurred in the amount of Bence Jones protein excretion; these changes were especially evident when 75 mg of prednisone were given daily for 7 days as part of a monthly chemotherapeutic regimen. Within the 7-day period seven patients showed essentially no decrease (<25%), whereas 13 and 9 patients had a moderate decrease (25-75%) or a marked decrease (>75%), respectively, in Bence Jones proteinuria as compared to pre-treatment values. The decrease in excretion of Bence Jones protein during this period was attributed mainly to corticosteroid therapy because of the transient nature of the response in most patients and the lack of such response in three patients when the hormone was omitted. Biosynthetic studies were performed to determine in vitro the effect of corticosteroids on Bence Jones protein synthesis. Plasma cells obtained from the bone marrow of 13 patients were incubated in a growth medium containing (14)C-labeled lysine and isoleucine and prednisone in concentrations up to 240 mug/ml, and the amount of Bence Jones protein synthesized was determined immunochemically. No differences in viability were apparent between untreated and prednisone-treated cells. The type of response exhibited by an individual patient in the percent decrease of Bence Jones protein excreted after 7 days of prednisone treatment was comparable to the percent decrease in newly-synthesized Bence Jones protein secreted by tumor cells when cultured in the presence of prednisone at a concentration of 120 mug/ml. The marked differences in the capacity of corticosteroids to affect Bence Jones protein synthesis appear to reflect a biochemical heterogeneity among plasma cell neoplasms.

Adult↗

Daily illumination exposure and melatonin: influence of ophthalmic dysfunction and sleep duration.

BACKGROUND: Ocular pathology lessens light's efficacy to maintain optimal circadian entrainment. We examined whether ophthalmic dysfunction explains unique variance in melatonin excretion of older adults over and above the variance explained by daily illumination, medical, and sociodemographic factors. We also examined whether ophthalmic dysfunction influences relationships between ambient illumination and melatonin. METHODS: Thirty older adults (mean age = 69 years; Blacks = 42% and Whites = 58%) of both genders participated in the study. Demographic and health data were collected at baseline. Participants underwent eye exams at SUNY Downstate Medical Center, wore an actigraph to monitor illumination and sleep, and collected urine specimens to estimate aMT6s concentrations. RESULTS: Hierarchical regression analysis showed that illumination factors explained 29% of the variance in aMT6s mesor. The proportion of variance explained by ophthalmic factors, sleep duration, and race was 10%, 2%, and 2%, respectively. Illumination factors explained 19% of the variance in aMT6s acrophase. The proportion of variance explained by ophthalmic factors, sleep duration, and race was 11%; 17%; and 2%, respectively. Controlling for sleep duration and race reduced the correlations between illumination and melatonin, whereas controlling for ophthalmic factors did not. CONCLUSION: Ophthalmic exams showed that elevated intraocular pressure and large cup-to-disk ratios were independently associated with earlier melatonin timing. Lower illumination exposure also had independent associations with earlier melatonin timing. Conceivably, ophthalmic and illumination factors might have an additive effect on the timing of melatonin excretion, which in turn might predispose individuals to experience early morning awakenings.

Journal Article↗

Evaluation of urinary PrPSc as a diagnostic test for sporadic, variant, and familial CJD.

Previously collected urine specimens from 100 patients referred to the UK National CJD Surveillance Unit as suspected cases of Creutzfeldt-Jakob disease (CJD) were analyzed, testing for abnormal prion protein (PrP(Sc)). In this context, the test had a low sensitivity and was not completely specific for CJD. Additionally, the proteins detected by this assay were not PrP(Sc) but appeared to be immunoglobulins.

Antibody Specificity↗

[13C]Nandrolone excretion in trained athletes: interindividual variability in metabolism.

BACKGROUND: Nandrolone is one of the most abused anabolic steroids, and its use in doping is increasing, as revealed by numerous positive cases during recent years in various sports. Different authors have reported the possible natural production of nandrolone metabolites in humans, and some of these authors argued that exhaustive exercise could increase nandrolone production in the body or induce dehydration and consequently lead to an increase of nandrolone metabolites in urine. METHODS: Volunteers (n = 22) ingested two 25-mg doses of [(13)C]nandrolone at 24-h intervals and collected urine specimens for 5 days. The labeled nandrolone metabolites 19-norandrosterone and 19-noretiocholanolone were identified and quantified by gas chromatography-mass spectrometry. RESULTS: Interindividual variability was observed in nandrolone excretion patterns and kinetics, as well as for the noretiocholanolone:norandrosterone ratio. The amounts of nandrolone metabolites measured at the excretion peak varied between 1180 and 38 661 microg/L for norandrosterone and 576 and 12 328 microg/L for noretiocholanolone. At the end of the excretion period, the noretiocholanolone:norandrosterone ratio was sometimes >1. The analysis of numerous spot-urine samples allowed the determination of an acceptable correlation between urinary creatinine and specific gravity for placebo- and steroid-treated individuals: y = 0.0052ln(x) + 1.0178 (r(2) = 0.8142) and y = 0.0068ln(x) + 1.0172 (r(2) = 0.7730), respectively. CONCLUSIONS: The excretion kinetics and patterns of labeled nandrolone show interindividual variability. More investigations are currently underway to estimate the influence of exhaustive exercises on excretion of labeled nandrolone metabolites in urine.

Adult↗

Desmopressin and low-dose ACTH test in rheumatoid arthritis.

OBJECTIVE: To ascertain whether a different regulation and sensitivity of the hypothalamic-pituitary-adrenal axis exists and whether a type of cortisol resistance is present in rheumatoid arthritis (RA) patients, a chronic disease in whose pathogenesis modifications of the steroid milieu are involved. DESIGN: We studied the basal and dynamic response of ACTH and adrenal steroids to various stimuli acting on the hypophysis or directly on the adrenal gland. METHODS: We studied ten RA patients (39.8 +/- 7.4 (S.D.) years), defined according to the American Rheumatism Association, and seven healthy control patients (34.1 +/- 9.6 (S.D.) years). All subjects underwent testing, in random order, with placebo, desmopressin (DDAVP) (10 microg i.v.), ovine corticotropin-releasing hormone (oCRH) (1 microg/kg body weight) and low-dose ACTH (5 microg i.v.), during the follicular phase of two different menstrual cycles. Blood samples were collected at different times for ACTH and adrenal steroids assay. Baseline estradiol (E2), testosterone and IGF-I levels were also evaluated. All subjects collected urine specimens for 24 h urine free cortisol (UFC). RESULTS: No difference in E2, testosterone or UFC was found between RA patients and controls. IGF-I levels were significantly (P < 0.01) lower in RA patients (110.6 +/- 6.4 microg/l) than in controls (207.0 +/- 37.9 microg/l). Mean baseline dehydroepiandrosterone (DHEA) and delta4-androstenedione levels of the four tests were significantly (P < 0.05) lower in RA patients than in controls. In RA, a negative correlation was found between mean DHEA levels, class of disease (r = -0.67, P < 0.05) and erythrocyte sedimentation rate (r = -0.63, P < 0.05). After placebo no difference in ACTH and cortisol area under curves (AUCs) was found between RA patients and controls. After DDAVP no cortisol or ACTH response was found in RA patients, while a significant (P < 0.05) ACTH release was found in controls. Only in RA patients was DDAVP able to induce a significant (P < 0.01) DHEA increase. After oCRH a similar significant response in ACTH (P < 0.05), cortisol (P < 0.01), and DHEA (P < 0.01) was found in both groups. After low-dose ACTH, a similar significant (P < 0.01) cortisol response was found in both RA patients and controls; indeed in RA patients DHEA AUC (2196.0 +/- 321.8 nmol/l per 90 min) was significantly lower (P < 0.01) than DHEA AUC (4280.8 +/- 749.0 nmol/l per 90 min) in controls. A similar significant (P < 0.01), though not abnormal, 17-hydroxyprogesterone response to ACTH was found in both groups. CONCLUSIONS: Our study underlines reduced adrenal steroid and IGF-I levels, but not the previously described cortisol resistance in RA patients; it shows that baseline and dynamic cortisol levels are 'normal' but inadequate in the setting of a sustained inflammatory disease like RA. The reduced basal and low-dose ACTH-induced DHEA levels could reflect both a reduced sensitivity of the adrenal gland to exogenous corticotropin and a decreased steroid synthesis due to a partial adrenal enzymatic defect (P450 17,20 lyase).

Adrenocorticotropic Hormone↗

Chlamydia trachomatis and Neisseria gonorrhoeae infections among men and women entering California prisons.

OBJECTIVE: We estimated the prevalence of Chlamydia trachomatis and Neisseria gonorrhoeae infection among newly arriving inmates at 6 California prisons. METHODS: In this cross-sectional study in 1999, urine specimens collected from 698 men aged 18 to 25 years and 572 women aged 18 years or older were tested at intake for C trachomatis and N gonorrhoeae using ligase chain reaction. An analysis of demographic and arrest-related correlates of C trachomatis and N gonorrhoeae infection was performed. RESULTS: The overall C trachomatis prevalence was 9.9% (95% CI=7.8%, 12.3%) among men aged 18 to 25 years, 8.9% (95% CI = 2.9%, 22.1%) among women aged 18 to 25 years, and 3.3% (95% CI=2.0%, 5.1%) among women overall. Three N gonorrhoeae cases were detected with an overall prevalence of 0.24% (95% CI=0.05%, 0.69%). CONCLUSIONS: The prevalence of C trachomatis infection at entry to California prisons, especially among young female and male inmates, was high, which supports routine screening at entry into prison. In addition, screening in a jail setting where most detainees are incarcerated before entry into the prison setting may provide an excellent earlier opportunity to identify these infections and treat disease to prevent complications and burden of infection in this high-risk population.

Adolescent↗

Two studies of compliance with daily prophylaxis in rheumatic fever patients in Iowa.

We determined the penicillin compliance rates in school year 1974-75 for 235 children with rheumatic fever through analysis of urine specimens collected at school. Patients relatively low on compliance during the first year of sampling were divided into two equal size groups for a second year sampling study. One-half (N = 44) of these second year study patients and their parents were readvised regarding the need for penicillin prophylaxis and one-half (N = 44) served as controls. We related eductional data, behavioral ratings, and family information to compliance rates in both study years. The overall compliance for the children during the initial sampling year was 64.6 per cent. Factors found to be significantly related to compliance in year one were: 1) male sex, 2) large sibship, 3) increased number of behavior problems, 4) relatively poor academic progress, 5) lower maternal educational level, and 6) recurrence of rheumatic fever. Compliance rates for reinforced patients did not differ significantly from controls during the second year sampling.

Adult↗

Enzyme immunoassay and direct immunofluorescence for detection of Chlamydia trachomatis antigen in male first-void urine.

First-void urine specimens, collected from 309 military recruits, 246 male adolescent gymnasium students and 194 patients consulting venereal disease clinics, were studied for the presence of Chlamydia trachomatis with the use of antigen detection tests--two enzyme immunoassays (EIA) and a direct immunofluorescence test (DIF; Syva MicroTrak). Urethral swabs were collected when discrepancies between the EIA and DIF tests were detected. The patient was regarded as positive when the culture result was positive or when two antigen detection tests corraborated one another. The Syva MicroTrak EIA and DIF tests were more sensitive than the Orion EIA, i.e. 98.5%, 99.2% and 74%, respectively. This was true when testing both low- and high-risk groups, with a prevalence of chlamydial infection ranging from 0.4% to 58.6%. All three tests were highly specific. The positive predictive values for the Syva MicroTrak EIA, the DIF and the Orion EIA were 99.2%, 100% and 100%, respectively and the negative predictive values 99.8%, 99.8% and 94.8%, respectively.

Adolescent↗

Progesterone and human chorionic gonadotrophin in serum and pregnandiol in urine in threatened abortion.

Progesterone and human chorionic gonadotrophin (HCG) in serum and pregnandiol in urine were measured in 64 patients admitted to hospital because of threatened abortion. Blood samples were taken and urine specimens collected at regular intervals during admission and after discharge during the rest of the pregnancy. A reference range was worked out for each hormone based on the hormone values obtained from the pregnancies proceeding to term. The predictive significance of values within and below the reference range was determined, for the initial sample and for serial samples. An association between hormone levels and outcome of pregnancy was observed but it is concluded that both single and serial determinations of progesterone and pregnandiol and serial determinations of HCG are unsatisfactory for the evaluation of threatened abortion. However, an initial progesterones value below the reference range and HCG values below 10,000 mIU/ml between the 8th and 15th week of pregnancy was in every case always followed by spontaneous abortion. A hormonal test of fetoplacental origin is recommended for monitoring threatened abortion.

Abortion, Threatened↗

Efficacy of two commercial products for altering urine drug test results.

OBJECTIVE: We have become aware of several commercial products that, when orally ingested, will purportedly not only eliminate "toxins" from a person's system, but will also correct any urinary imbalances caused by excessive water consumption. METHOD: Unblinded study of one volunteer subject, tested weekly x 4 for 24-hour urine elimination of test drug under conditions of control, control plus 1200 mL water, Quick Flush', and Eliminator. RESULTS: Each of the treatment protocols studied caused reductions of drug or metabolite concentrations as measured by gas chromatography-mass spectrometry in urine specimens collected up to 24 hours after ingestion of amphetamine, 9-carboxy-11-nor-delta-9-THC, benzoylecgonine, or codeine, yet the radioimmunoassay screening results demonstrated very little effect. Water alone was approximately as effective as the two commercial products in reducing the metabolite level. None of the treatment protocols employed in this study altered urinary pH, specific gravity, or creatinine concentration outside the normally accepted physiological range. CONCLUSIONS: Attempts to conceal drug abuse by water dilution are most likely to play a substantial role when concentrations are at or near the detection threshold for a particular assay such as the terminal stages of drug eliminations.

Amphetamine↗

Schistosomiasis--an unusual cause of ureteral obstruction: a case history and perspective.

A male, 32 years of age, presented with dysuria and abdominal pain, but no gross hematuria. He emigrated three years earlier from Somalia, East Africa, and was currently employed as a poultry processor in a rural Wisconsin community. The patient denied any trauma, sexual activity, or family history of significant illness. Abdominal and genitourinary exams were normal with negative tests for gonococcus and chlamydia. Urinalysis demonstrated microhematuria. A urogram and retrograde pyelogram revealed a mildly dilated right ureter down to the ureterovesical junction. Cystoscopy showed punctate white lesions on the bladder urothelium. Ureteroscopy was used to biopsy abnormal tissue in the distal ureter and bladder. Biopsy tissue demonstrated deposits of Schistosoma haematobium eggs. No ova were seen in collected urine specimens. The patient was successfully treated with praziquantel and will be monitored for sequelae of the disease. Schistosomiasis (Bilharziasis) can be expected to be seen with increasing frequency in the United States with the continuing influx of immigrants and refugees, as well as the return of travelers and soldiers from endemic areas. While no intermediate snail host exists for the transmission of Schistosoma sp. in the United States, the continued importation of exotic animals including snails from Africa, as well as the ability of schistosomes to shift host species warrants concern. Additionally, increasing disease associated with non-human bird schistosomes of the same genus seen in the midwestern United States is occurring throughout Europe. One should be aware that praziquantel may not always be available or effective in the treatment of schistosomiasis. It behooves the practicing clinician to remain updated on the status of this widespread zoonosis.

Adult↗

Postal urine specimens: are they a feasible method for genital chlamydial infection screening?

BACKGROUND: A United Kingdom (UK) screening programme for Chlamydia trachomatis has recently been announced. Pilot projects involving the opportunistic testing of women attending health facilities are due to commence in several sites. There is a danger that this approach will fail to obtain adequate population coverage. The alternative--true systematic population screening--is generally assumed to be unfeasible. Studies in Denmark using postal urine specimens have challenged this assumption. No such studies have been reported from the UK. AIM: To assess the potential of urine specimens sent by post as the basis for a UK population screening strategy for genital chlamydial infection. METHOD: Two hundred patients (100 men, 100 women) aged 18 to 45 years were randomly sampled from the list of one urban group practice. Subjects were mailed an explanatory letter, a urine sample container, a sexual lifestyle questionnaire, and a prepaid return envelope. Non-responders were contacted by telephone; persistent non-responders were visited at home. Samples were tested for Chlamydia by DNA amplification and enzyme immunoassay. RESULTS: Sixty-four (32%) subjects were no longer living at their GP registered address. Of the remaining 136, 126 (93%) responded to the survey and 113 (83%) accepted the request for a urine sample and completed a questionnaire. Acceptance rates were similar for men and women and across age groups. Four samples (3%) were Chlamydia positive. CONCLUSION: Home mailed urine specimen collection in conjunction with a self-completed postal questionnaire is feasible. This could provide a viable basis both for determining population Chlamydia prevalence and for a UK Chlamydia population screening strategy. Overall cost effectiveness of such a strategy will depend on the cost of the test used. Comparative performance characteristics of the different currently available tests in this setting have yet to be fully determined.

Adolescent↗

Occurrence and antibiotic sensitivity of Enterobacteriaceae isolated from a group of Jordanian patients with community acquired urinary tract infections.

The type and antibiotic sensitivity of urinary tract pathogens may differ in various communities. Of 207 isolates recovered from midstream urine specimens collected from a group of patients with community acquired urinary tract infections (UTI), 86% were species of Enterobacteriaceae. The most frequently recovered pathogens were Escherichia coli (82%), Klebsiella spp. (7.3%), Proteus spp. (6.2%), Enterobacter spp. (3.4%) and Citrobacter spp. (1.1%). High rates of resistance were found against ampicillin (95%), tetracycline (86%), carbenicillin (84%), trimethoprim/sulphamethoxazole (48%), and amoxycillin/clavulanic acid (45%). For the antibiotics tobramycin, aztreonam, ceftriaxone and gentamicin 7% of the isolates were resistant, while resistance varied from 9-18% for amikacin, ciprofloxacin, norfloxacin, nalidixic acid and cefuroxime. The incidence of UTI caused by Enterobacteriaceae was three times higher in females than in males, particularly in young and middle age groups (< or = 19 and 20-39 years).

Adolescent↗

Recent measles viral activity in Uruguai: serological and genetic approaches.

In the summer 1999, a measles outbreak occurred in Uruguai. During this outbreak 58 cases were recorded, 36 of which were laboratory confirmed as positive for measles virus (MV) IgM. The cases occurred in touristic places (Montevideo and Maldonado) predominantly among health facilities and tourist service personnel. Urine specimens collected between days 1 and 4 after the onset of the rash from seven cases were analyzed by reverse transcription-polymerase chain reaction (RT-PCR) and nested PCR with primers specific for the carboxyl-terminal region of the nucleoprotein (N) gene. Three of these specimens/cases were positive for MV. Sequencing of 300 nucleotides (nt) of PCR products corresponding to a part of the carboxyl-terminal region of the MV N gene detected in these specimens MV of D6 genotype. The same nucleotide sequences and the same genotype were also previously observed for MV isolates from the 1997 epidemic in Brazil and the 1998 epidemic in Argentina, demonstrating that the D6 genotype was, and may be still circulating in South America.

Adult↗