Lung cancer risk test trial: study design, participant baseline characteristics, bronchoscopy safety, and establishment of a biospecimen repository.

BACKGROUND: The Lung Cancer Risk Test (LCRT) trial is a prospective cohort study comparing lung cancer incidence among persons with a positive or negative value for the LCRT, a 15 gene test measured in normal bronchial epithelial cells (NBEC). The purpose of this article is to describe the study design, primary endpoint, and safety; baseline characteristics of enrolled individuals; and establishment of a bio-specimen repository. METHODS/DESIGN: Eligible participants were aged 50-90 years, current or former smokers with 20 pack-years or more cigarette smoking history, free of lung cancer, and willing to undergo bronchoscopic brush biopsy for NBEC sample collection. NBEC, peripheral blood samples, baseline CT, and medical and demographic data were collected from each subject. DISCUSSION: Over a two-year span (2010-2012), 403 subjects were enrolled at 12 sites. At baseline 384 subjects remained in study and mean age and smoking history were 62.9 years and 50.4 pack-years respectively, with 34% current smokers. Obstructive lung disease (FEV1/FVC <0.7) was present in 157 (54%). No severe adverse events were associated with bronchoscopic brushing. An NBEC and matched peripheral blood bio-specimen repository was established. The demographic composition of the enrolled group is representative of the population for which the LCRT is intended. Specifically, based on baseline population characteristics we expect lung cancer incidence in this cohort to be representative of the population eligible for low-dose Computed Tomography (LDCT) lung cancer screening. Collection of NBEC by bronchial brush biopsy/bronchoscopy was safe and well-tolerated in this population. These findings support the feasibility of testing LCRT clinical utility in this prospective study. If validated, the LCRT has the potential to significantly narrow the population of individuals requiring annual low-dose helical CT screening for early detection of lung cancer and delay the onset of screening for individuals with results indicating low lung cancer risk. For these individuals, the small risk incurred by undergoing once in a lifetime bronchoscopic sample collection for LCRT may be offset by a reduction in their CT-related risks. The LCRT biospecimen repository will enable additional studies of genetic basis for COPD and/or lung cancer risk. TRIAL REGISTRATION: The LCRT Study, NCT 01130285, was registered with Clinicaltrials.gov on May 24, 2010.

HIV awareness and condom use among female sex workers in Afghanistan: implications for intervention.

There is little information about HIV awareness or condom use among female sex workers (FSWs) in Afghanistan. The purpose of this cross-sectional study was to assess HIV awareness, knowledge, and condom use among FSWs in three Afghan cities. FSWs residing in Jalalabad, Kabul, and Mazar-i-Sharif were recruited through outreach programs and completed an interviewer-administered questionnaire and rapid tests for hepatitis B surface antigen, HIV, syphilis, and hepatitis C virus. Logistic regression identified factors associated with HIV awareness, comprehensive HIV knowledge (knowledge that HIV cannot be detected by sight, that condoms prevent HIV, and rejection of local misconceptions about HIV transmission), and consistent condom use (use with every sex act) with clients in the last six months. Of 520 participants, 76.9% had no formal education and 37.7% lived outside Afghanistan in the last five years. Nearly half (44.2%) were aware of HIV but, of these, only 17.4% (N = 40) had comprehensive HIV knowledge. There were significant differences by site; FSWs in Jalalabad were more likely to be aware of HIV but FSWs in Kabul were more likely to have correct HIV knowledge and use condoms consistently with clients. Consistent client condom use was reported by 11.5% (N = 60) and was independently associated with having more clients per month (AOR = 1.99, 95% CI: 1.04-3.81). In conclusion, comprehensive HIV knowledge and consistent condom use with clients are low among Afghan FSWs in these cities. Efforts to reach this population should focus on relaying accurate information and expanding condom use with clients.

Implications of hepatitis C viremia vs. antibody alone on transmission among male injecting drug users in three Afghan cities.

OBJECTIVES: To assess differences between injecting drug users (IDUs) with hepatitis C virus (HCV) viremia and IDUs with HCV antibody (Ab) or no evidence of prior infection in three Afghan cities. METHODS: IDUs in Hirat, Jalalabad, and Mazar-i-Sharif completed questionnaires and rapid testing for blood-borne infections including HCV Ab. HCV Ab was confirmed with a recombinant immunoblot assay (RIBA); RIBA-positive specimens underwent reverse transcriptase polymerase chain reaction (RT-PCR) for HCV. Risk behaviors associated with viremia were assessed with site-controlled ordinal regression analysis. RESULTS: Of 609 participants, 223 (36.6%) had confirmed HCV Ab. Of 221 with serum available for PCR evaluation, 127 (57.5%) were viremic. HCV viremia prevalence did not differ by site (range 41.7-59.1%; p=0.52). Among all IDUs, in age and site-controlled ordinal regression analysis, HCV was independently associated with HIV co-infection (adjusted odds ratio (AOR) 7.16, 95% confidence interval (CI) 4.41-11.64), prior addiction treatment (AOR 1.95, 95% CI 1.57-2.42), ever aspirating and re-injecting blood (AOR 1.62, 95% CI 1.18-2.23), prior incarceration (AOR 1.60, 95% CI 1.04-2.45), and sharing injecting equipment in the last 6 months (AOR 1.35, 95% CI 1.02-1.80). CONCLUSION: HCV viremia was present in many participants with prior HCV infection and was associated with some injecting risk behaviors, indicating a substantial risk for transmission. Current harm reduction programs should aim to improve HCV awareness and prevention among IDUs in Afghanistan as a matter of urgency.

Prevalence of HIV, hepatitis B and hepatitis C and associated risk behaviours amongst injecting drug users in three Afghan cities.

BACKGROUND: HIV amongst injecting drug users (IDUs) has been described in Kabul but little data exists for other Afghan cities. We assessed HIV, hepatitis B virus (HBV), and C virus (HCV) prevalence and associated risk behaviours amongst IDUs in Hirat, Jalalabad, and Mazar-i-Sharif, Afghanistan. METHODS: Consented participants reporting injecting drugs within the previous 6 months completed interviewer-administered questionnaires and testing for HIV, hepatitis C antibody (HCV Ab), and hepatitis B surface antigen (HBsAg). Logistic regression was used to determine characteristics associated with each infection. RESULTS: Of 623 participants, most (98.7%) were male. Prevalence of HIV, HCV, and HBV was 1.8% (95% CI: 0.88-3.2), 36.0% (95% CI: 33-41), and 5.8% (95% CI: 3.9-7.6), respectively. All HIV cases and highest HCV prevalence were detected in Hirat; HBV prevalence was highest in Jalalabad. Amongst male IDUs, 62.9% had been imprisoned, of whom 17.2% (n=66) injected in prison. High risk behaviours were common; 30.2% reported needle sharing in the last 6 months, 23.1% reported sex with another male, and 50.4% reported paying females for sex. Behaviours varied significantly by site; generally, Hirat participants reported fewer sexual risk behaviours. Sex with other males was negatively associated with both HBV and HCV in multivariate logistic regression analysis; no injecting behaviours were associated with both HBV and HCV. CONCLUSIONS: Whilst HIV prevalence is low, HCV prevalence and high risk behaviours were common in these populations. Regional variations should be considered in programming to prevent transmission of HIV and viral hepatitis amongst IDUs in Afghanistan.

Contraceptive utilization and pregnancy termination among female sex workers in Afghanistan.

BACKGROUND: To determine the prevalence and correlates of prior pregnancy termination and unmet need for contraception among female sex workers (FSWs) in Afghanistan. METHODS: FSWs in Jalalabad, Kabul, and Mazar-i-Sharif were recruited between June 2006 and December 2007 through outreach programs. Participants completed an interviewer-administered survey describing demographics, behaviors associated with risk of sexually transmitted infections (STIs) and unplanned pregnancy, and medical history. Correlates of prior pregnancy termination and current unmet need for contraception were assessed with logistic regression analysis, controlling for site. RESULTS: Of 520 FSWs, most (82.3%) had been pregnant at least once (mean 4.9 ± 2.7, range 1-17), among whom unplanned pregnancy (36.9%) and termination (33.2%) were common. Jalalabad participants were more likely to report both prior unplanned pregnancy (60.6% vs. 48.3% in Kabul or 20.7% in Mazar, p < 0.001) and prior termination (54.9% vs. 31.8% in Kabul or 26.8% in Mazar, p < 0.001). Most FSWs (90.0%) stated pregnancy was not currently desirable, and 85.2% were using contraception. Unmet need for contraception (14.7% of participants) was positively associated with having sold sex outside their city of residence (adjusted odds ratio [AOR] 1.88, 95% confidence interval [CI] 1.28-2.77) and inversely associated with illicit drug use (AOR 0.41, 95% CI 0.31-0.53). CONCLUSIONS: Although FSWs in Afghanistan report high rates of contraceptive use, unplanned pregnancy is common. Reproductive health services should be included in programming for FSWs to reduce unplanned pregnancies and to reduce HIV/STI risks.

HIV, hepatitis B, and hepatitis C prevalence and associated risk behaviors among female sex workers in three Afghan cities.

OBJECTIVE: To assess prevalence of HIV, syphilis, and hepatitis B (HBV) and C virus (HCV) and associated risk behaviors among female sex workers (FSWs) in three Afghan cities. DESIGN: Cross-sectional prevalence assessment. METHODS: Consented FSWs from Jalalabad, Kabul, and Mazar-i-Sharif completed an interviewer-administered questionnaire, pretest and posttest counseling, and rapid and confirmatory testing for HIV, HCV, HBV, and syphilis. Logistic regression was used to detect correlates associated with HBV infection. RESULTS: Of 520 participants, median age and age of initiating sex work were 29 and 23 years, respectively, and the median number of monthly clients was 12. Few FSWs reported ever having used illicit drugs (6.9%) or alcohol (4.7%). Demographic and risk behaviors varied significantly by enrollment site, with Kabul FSWs more likely to report sexually transmitted infection symptoms, longer sex work duration, and sex work in other cities. Prevalence of HIV was 0.19%, HCV was 1.92%, and HBV was 6.54%, with no cases of syphilis detected. HBV was independently associated with at least 12 clients monthly [adjusted odds ratio (AOR) = 3.15, 95% confidence interval (CI) 1.38-7.17], ever using alcohol (AOR = 2.61, 95%CI 1.45-4.69), anal sex (AOR = 2.42, 95%CI 1.15-5.08), and having children (AOR = 2.12, 95%CI 1.72-2.63) in site-controlled multivariable analysis. CONCLUSION: Although prevalence of HIV, HCV, and syphilis is currently low in these three Afghan cities, risky sexual practices were common and associated with HBV. Programming inclusive of voluntary testing for HIV, viral hepatitis, and sexually transmitted infections, hepatitis vaccination, substance abuse prevention, and condom promotion for both FSWs and clients should be pursued in Afghanistan.

Prevalence and correlates of syphilis and condom use among male injection drug users in four Afghan cities.

BACKGROUND: : Injecting drug use is increasing in Afghanistan but little is known about sexual risk behaviors and sexually transmitted infection (STI) prevalence among injection drug users (IDU). The purpose of this study is to assess prevalence and correlates of syphilis and condom use with female sex workers (FSWs) among male IDUs in Hirat, Jalalabad, Kabul, and Mazar-i-Sharif, Afghanistan. METHODS: : Participants in this cross-sectional study completed an interviewer-administered questionnaire and serologic testing for syphilis between June 2005 and January 2008. Factors associated with syphilis condom use with FSWs were assessed with site-controlled logistic regression analysis. RESULTS: : Of 1078 male IDUs, most (90.3%) reported prior sexual experience, of whom 27.6% reported any condom use. Sexual experiences with FSWs (58.1%) and men or boys (25.7%) were common, although prior condom use with FSWs (32.6%) or male partners (10.8%) was relatively rare. Few reported having a lifetime STI diagnosis (6.3%, n = 68) or symptoms (10.4%, n = 110) in the last 6 months. Prevalence of syphilis was 3.72% (95% CI: 2.66%-5.06%) and varied significantly between sites ranging from 0% (Jalalabad) to 13.9% (Mazar-i-Sharif) (P < 0.001)). Syphilis was significantly associated with STI diagnosis (adjusted odds ratio [AOR] = 3.84) or sex with FSWs (AOR = 3.82) in the last 6 months, and with lower (≤6 years) educational level (AOR = 2.20). Prior condom use with FSWs was independently associated with living outside Afghanistan in the last decade (AOR = 5.52, 95% CI: 1.83-16.71), higher income (AOR = 2.03, 95% CI: 1.17-3.51), greater number of lifetime partners (AOR = 1.80, 95% CI: 1.32-2.45), and younger age (AOR = 0.985, 95% CI: 0.973-0.998). CONCLUSIONS: : Although prevalence of syphilis and condom use varied significantly by site, high levels of risky sexual behavior were common, and consistent condom use was rare among IDUs in Afghanistan. Harm reduction programming should incorporate sexual risk reduction and condom promotion and distribution in Afghan cities.

The effectiveness of microballoon angioplasty in treating middle cerebral artery occlusion beyond the bifurcation.

BACKGROUND AND PURPOSE: PTA effectively treats vasospasm and arterial disease in peripheral, coronary, and large proximal cerebral vessels but rarely reaches small vessels like the distal MCA beyond the bifurcation. Our purpose was to evaluate the safety and efficacy of PTA for distal MCA occlusions in acute ischemic stroke. MATERIALS AND METHODS: Seven patients had strokes in branches of their MCAs. Following diagnostic angiography, all received microballoon angioplasty and various combinations of IA alteplase (rtPA), abciximab, and/or nitroglycerin. Two also underwent stent placement. Comprehensive retrospective review of the patients' records was performed. Patients' NIHSS scores were reassessed before discharge. Recanalization was evaluated by angiography after treatment and at follow-up. RESULTS: PTA was successfully performed in 7 patients without treatment-associated intracerebral hemorrhage. Two patients received distal MCA angioplasty as a secondary intervention: 1 following failed treatment with a Merci retriever and the other after successful removal of proximal clot with a Merci retriever. One patient did not recover from the initial ischemic event despite an excellent angiographic result. Complete recanalization (modified TIMI grade 4) was achieved in 4 patients and near-complete recanalization with mild flow deficit (modified TIMI grade 3), in 3 patients. CONCLUSIONS: PTA of the distal MCA with a microballoon is safe and effective for acute ischemic stroke. This case series demonstrates that endovascular treatment beyond the MCA bifurcation can dramatically reverse neurologic deficits.

Effects of duration of injection drug use and age at first injection on HCV among IDU in Kabul, Afghanistan.

BACKGROUND: Hepatitis C virus (HCV) prevalence is high among injection drug users (IDUs) in Afghanistan. Duration of injection and young age at first injection are common risk factors for HCV in IDU populations. The association of HCV with these time factors was analyzed. METHODS: Socio-demographic and drug use behavior information were collected. Participants had rapid testing for HCV with recombinant immunoblot assay confirmation. Modeling of non-linear associations was performed using fractional polynomial logistic regression. RESULTS: Among 459 male IDUs, age at first injection had a constant HCV risk (odds ratio (OR): 1.01 per year; 95% confidence interval (CI): 0.98-1.03), while each additional year of injection drug use had a significantly increased risk (OR: 4.72 per year, 95% CI: 2.92-7.66). HCV risk increased significantly with each additional year of injecting drug use by groups of injectors: young (< or =22 years, OR: 1.97; 95% CI: 1.27-3.07), middle (23-28 years, OR: 1.76; 95% CI: 1.28-2.43) and older (> or =29 years, OR: 7.56; 95% CI: 3.15-18.14). CONCLUSION: The probability of HCV infection increased markedly by duration of injection drug use and varied according to age at first injection. Drug counseling and educational efforts should be directed to older drug users who have not yet initiated injecting and to young IDUs to avert infection and reduce risky drug use behaviors.

Correlates of receptive and distributive needle sharing among injection drug users in Kabul, Afghanistan.

We describe receptive and distributive needle/syringe sharing among injection drug users (IDUs) in Kabul, Afghanistan. In this cross-sectional study, IDUs completed an interviewer-administered questionnaire. Logistic regression identified correlates of needle sharing in the last six months. Receptive and distributive sharing in the last six months were reported by 28.2% and 28.7% of participants, respectively, and were both independently associated with reported difficulty obtaining new syringes (Receptive sharing: AOR = 2.60, 95% CI: 1.66-4.06; Distributive: AOR = 1.56, 95% CI: 1.02-2.39). Receptive and distributive sharing are common among IDU in Kabul; scaling up availability of sterile, no-cost injecting equipment is urgently needed.

Genetic characterization of HIV-1 strains circulating in Kazakhstan.

To determine the HIV-1 genetic diversity in Kazakhstan, 85 blood samples from HIV-seropositive donors were collected between 2001 and 2003. The study population consisted of 91.8% injecting drug users (IDUs); the remainder was infected sexually or iatrogenically. A genomic region that included part of the polymerase gene was sequenced for all 85 samples, and from these, 6 samples were randomly selected for nearly full genome sequencing. Subtype A was the most common genetic form (94.1%), followed by CRF02_AG (4.7%) and subtype C (1.2%). All subtype A sequences clustered closely with samples from countries of the former Soviet Union (FSU). From these sequences, 47 (58.8%) presented the secondary protease inhibitor mutation V77I that has been linked to a genetic lineage in the FSU epidemic. In addition, most had the other 2 mutations that characterize the "V77I haplotype." All 6 nearly full-length sequences were subtype A and clustered with other FSU strains. The CRF02_AG strains from this population clustered with strains from Uzbekistan, reflecting the spread of the CRF02_AG epidemic in Central Asia. The HIV epidemic in Kazakhstan is predominantly in IDUs and is indigenous to the geographic region, and most of the strains are genetically similar to those circulating in the FSU and other parts of Central Asia.

Association between expatriation and HIV awareness and knowledge among injecting drug users in Kabul, Afghanistan: A cross-sectional comparison of former refugees to those remaining during conflict.

BACKGROUND: Little is known about human immunodeficiency virus (HIV) awareness among Afghan injecting drug users (IDUs), many of whom initiated injecting as refugees. We explored whether differences in HIV awareness and knowledge exist between Afghan IDUs who were refugees compared to those never having left Afghanistan. METHODS: A convenience sample of IDUs in Kabul, Afghanistan was recruited into a cross-sectional study through street outreach over a one year period beginning in 2005. Participants completed an interviewer-administered questionnaire and underwent voluntary counseling and testing for HIV, syphilis, hepatitis B surface antigen, and hepatitis C antibody. Differences in HIV awareness and specific HIV knowledge between IDU who lived outside the country in the last decade versus those who had not were assessed with logistic regression. RESULTS: Of 464 IDUs, 463 (99%) were male; median age and age at first injection were 29 and 25 years, respectively. Most (86.4%) had lived or worked outside the country in the past ten years. Awareness of HIV was reported by 46.1%; those having been outside the country in the last decade were significantly more likely to have heard of HIV (48.3% vs. 31.7%; OR = 2.00, 95% CI: 1.14 - 3.53). However, of those aware of HIV, only 38.3% could name three correct transmission routes; specific HIV knowledge was not significantly associated with residence outside the country. CONCLUSION: Accurate HIV knowledge among Afghan IDUs is low, though former refugees had greater HIV awareness. Reported high-risk injecting behavior was not significantly different between IDU that were refugees and those that did not leave the country, indicating that all Afghan IDU should receive targeted prevention programming.

HIV, hepatitis C, and hepatitis B infections and associated risk behavior in injection drug users, Kabul, Afghanistan.

Limited prevalence data for HIV, hepatitis B surface antigen (HBsAg), and hepatitis C virus (HCV) exist for Afghanistan. We studied a cross-sectional sample of adult injection drug users (IDUs) in Kabul, Afghanistan, from June 2005 through June 2006. Study participants completed interviewer-administered questionnaires and underwent testing for HIV, antibody to HCV, and HBsAg. Overall prevalences of HIV, HCV, and HBsAg were 3.0% (95% confidence interval [CI] 1.7%-5.1%), 36.6% (95% CI 32.2%-41.0%), and 6.5% (95% CI 4.2%-8.7%), respectively (N = 464). Among male IDUs (n = 463), risky behavior, including sharing syringes (50.4%), paying women for sex (76.2%), and having sex with men or boys (28.3%), were common. Needle sharing, injecting for > or = 3 years, and receiving injections from nonmedical providers were independently associated with increased risk for HCV infection. The high prevalence of risky behavior indicate that Kabul is at risk for an HIV epidemic. Scale-up of harm-reducing interventions is urgently needed.

Genetic forms of HIV Type 1 in the former Soviet Union dominate the epidemic in Azerbaijan.

A total of 125 strains collected in Azerbaijan between 1999 and 2002 from HIV seropositives were genetically classified. Of 84 strains classified using HMA, 91.6% were subtype A, 1.2% subtype B, and 7.1% untypeable. Of 41 strains analyzed using partial pol gene sequences, 90.2% were subtype A, 7.3% subtype B, and 2.4% CRF03_AB. Most sequenced A strains clustered with those circulating in countries of the former Soviet Union (FSU). Two of three sequenced B strains were from individuals who traveled to FSU clustering tightly with B strains from Nikolayev, Ukraine. CRF03_AB, characteristic of the 1996 epidemic in Kaliningrad, Russia, was sequenced from an individual who lived in Russia from 1995 until 2001. The HIV epidemic in Azerbaijan is concentrated in IDU and is closely connected to other such epidemics to the east based on genetics. Of the 41 sequenced strains, 95% were close genetic relatives of HIV strains in IDU networks in the FSU.

High HIV prevalence and risk factors among injection drug users in Tashkent, Uzbekistan, 2003-2004.

OBJECTIVES: To determine HIV prevalence and potential associations with sociodemographic and behavioral factors among injection drug users (IDUs) in Tashkent, Uzbekistan. METHODS: Subjects in this cross-sectional study provided sociodemographic and risk behavior data and were tested for HIV antibody with a saliva-based enzyme-linked immunosorbent assay (ELISA), followed by saliva-based Western blot confirmation. RESULTS: Among the 701 IDU subjects enrolled, 209 (29.8%) were diagnosed with HIV infection, HIV infection was more likely among those unemployed (adjusted odds ratio [AOR]=1.47); whose first drug of abuse was heroin (AOR=2.21) or opium poppy extract (AOR=1.61); with a prior history of hepatitis (AOR=1.39); and those who reported never using condoms (AOR=1.65). Independent risk factors associated with HIV infection were heroin as the first illicit drug of abuse, prior hepatitis, lack of condom use, and unemployment. CONCLUSIONS: Heroin use, sexual transmission, and high unemployment seem to play important roles in HIV transmission. Implementation of effective harm reduction strategies is critical to control the expansion of the HIV epidemic in this country as well as in this region.

Prevalence and correlates of human immunodeficiency virus infection among female sex workers in Tashkent, Uzbekistan.

OBJECTIVES/GOAL: To assess prevalence of and correlates to human immunodeficiency virus (HIV) infection among female sex workers in Tashkent, Uzbekistan. STUDY DESIGN: Women participating in this cross-sectional study completed a questionnaire and HIV testing between April 2003 and March 2004. Logistic regression analyses determined correlation of variables to HIV infection. RESULTS: Of 448 women, 10% (45) were HIV infected, which was associated with ever injecting drugs (AOR = 20.20; 95% confidence interval [CI], 7.69-53.07), street-based sex work (AOR = 4.52; 95% CI, 1.84-11.12), exchanging sex for drugs (AOR = 4.74; 95% CI, 1.84-12.18), and more sexually transmitted infection treatments in the preceding 3 months (AOR = 2.43; 95% CI, 1.14-5.17). CONCLUSIONS: Although injection drug use is the strongest correlate to HIV infection, sexual risk behaviors are independently related and should receive focus in prevention efforts targeted to this population.

Molecular evaluation of antibiotic susceptibility of Tropheryma whipplei in axenic medium.

OBJECTIVES AND METHODS: Whipple's disease is a rare multisystem chronic infection, involving the intestinal tract as well as various other organs. Tropheryma whipplei is a slow-growing facultative intracellular bacterium that remains poorly understood. In vitro antibiotic susceptibility testing has previously been assessed in cells using a real-time quantitative PCR assay. In this study, we have evaluated the antibiotic susceptibility of three strains of T. whipplei grown in axenic medium using the same assay. RESULTS: The active compounds in axenic medium were doxycycline, macrolide compounds, penicillin G, streptomycin, rifampicin, chloramphenicol, thiamphenicol, teicoplanin, vancomycin, amoxicillin, gentamicin, aztreonam, levofloxacin and ceftriaxone, with MICs in the range 0.06-1 mg/L. Cefalothin was less active, with MICs in the range 2-4 mg/L. We found that co-trimoxazole was active with MICs in the range 0.5-1 mg/L, and sulfamethoxazole alone was active with MICs in the range 0.5-1 mg/L. MICs of trimethoprim varied from 64-128 mg/L. CONCLUSIONS: Co-trimoxazole was effective in vitro, but this activity was due to sulfamethoxazole alone. These results were in accordance with the fact that T. whipplei does not contain the encoding gene for dihydrofolate reductase, the target for trimethoprim.

Randomized comparison of intra-arterial and intravenous thrombolysis in a canine model of acute basilar artery thrombosis.

We compared the rates of recanalization cerebral infarct and hemorrhage between intra-arterial (i.a.) reteplase and intravenous (i.v.) alteplase thrombolysis in a canine model of basilar artery thrombosis. Thrombosis was induced by injecting a clot in the basilar artery of 13 anesthetized dogs via superselective catheterization. The animals were randomized in a blinded fashion, 2 h after clot injection and verification of arterial occlusion, to receive i.v. alteplase 0.9 mg/kg over 60 min and i.a. placebo, or i.a. reteplase 0.09 units/kg over 20 min, equivalent to one-half the alteplase dose, and i.v. placebo. Recanalization was studied for 6 h after treatment with serial angiography; the images were later graded in a blinded fashion. Blinded interpretation of postmortem MRI was performed to assess the presence of brain infarcts and/or hemorrhage. At 3 h after initiation of treatment, partial or complete recanalization was observed in one of six dogs in the i.v. alteplase group and in five of seven in the i.a. reteplase group (P = 0.08). At 6 h, no significant difference in partial or complete recanalization was observed between the groups (two of six vs. five of seven; P = 0.20). Postmortem MRI revealed infarcts in four of six animals treated with i.v. alteplase and three of seven treated with i.a. reteplase (P = 0.4). Intracerebral hemorrhage was more common in the i.v. alteplase group (four of six vs. none of seven; P = 0.02). This study thus suggests that i.a. thrombolysis affords a recanalization rate similar to that of i.v. thrombolysis, but with a lower rate of intracerebral hemorrhage.

Molecular evaluation of antibiotic susceptibility: Tropheryma whipplei paradigm.

Tropheryma whipplei, the agent of Whipple's disease, grows fastidiously only in cell cultures without plaque production, and only three strains have been passaged. The formation of bacterial clumps in the supernatant precludes enumeration of viable bacteria and MIC determination. We evaluated the bacteriostatic effects of fluoroquinolones against two T. whipplei isolates by measuring the inhibition of the DNA copy number increase by real-time quantitative PCR. The analysis of the T. whipplei genome database allowed the identification not only of the gyrA gene but also the parC gene encoding the alpha subunit of the natural fluoroquinolone targets DNA gyrase (GyrA) and topoisomerase IV (ParC), respectively. The parC gene was detected in actinobacteria for the first time. High ciprofloxacin MICs (4 and 8 micro g/ml) were correlated with the presence in T. whipplei GyrA and ParC sequences with an alanine residue at positions 83 and 80 (Escherichia coli numbering), respectively. Alanines at these positions have previously been associated with increased fluoroquinolone resistance in E. coli and mycobacteria. However, the MIC of levofloxacin was low (0.25 micro g/ml). The same T. whipplei GyrA and ParC sequences were found in two other cultured strains and in nine uncultured tissue samples from Whipple's disease patients, allowing one to speculate that T. whipplei is naturally relatively resistant to fluoroquinolones.