Bacterial vaginosis associated with increased risk of female-to-male HIV-1 transmission: a prospective cohort analysis among African couples.

BACKGROUND: Bacterial vaginosis (BV), a disruption of the normal vaginal flora, has been associated with a 60% increased risk of HIV-1 acquisition in women and higher concentration of HIV-1 RNA in the genital tract of HIV-1-infected women. However, whether BV, which is present in up to half of African HIV-1-infected women, is associated with an increase in HIV-1 transmission to male partners has not been assessed in previous studies. METHODS AND FINDINGS: We assessed the association between BV on female-to-male HIV-1 transmission risk in a prospective study of 2,236 HIV-1-seropositive women and their HIV-1 uninfected male partners from seven African countries from a randomized placebo-controlled trial that enrolled heterosexual African adults who were seropositive for both HIV-1 and herpes simplex virus (HSV)-2, and their HIV-1-seronegative partners. Participants were followed for up to 24 months; every three months, vaginal swabs were obtained from female partners for Gram stain and male partners were tested for HIV-1. BV and normal vaginal flora were defined as a Nugent score of 7-10 and 0-3, respectively. To reduce misclassification, HIV-1 sequence analysis of viruses from seroconverters and their partners was performed to determine linkage of HIV-1 transmissions. Overall, 50 incident HIV-1 infections occurred in men in which the HIV-1-infected female partner had an evaluable vaginal Gram stain. HIV-1 incidence in men whose HIV-1-infected female partners had BV was 2.91 versus 0.76 per 100 person-years in men whose female partners had normal vaginal flora (hazard ratio 3.62, 95% CI 1.74-7.52). After controlling for sociodemographic factors, sexual behavior, male circumcision, sexually transmitted infections, pregnancy, and plasma HIV-1 RNA levels in female partners, BV was associated with a greater than 3-fold increased risk of female-to-male HIV-1 transmission (adjusted hazard ratio 3.17, 95% CI 1.37-7.33). CONCLUSIONS: This study identified an association between BV and increased risk of HIV-1 transmission to male partners. Several limitations may affect the generalizability of our results including: all participants underwent couples HIV counseling and testing and enrolled in an HIV-1 prevention trial, and index participants had a baseline CD4 count ≥ 250 cells/mm³ and were HSV-2 seropositive. Given the high prevalence of BV and the association of BV with increased risk of both female HIV-1 acquisition and transmission found in our study, if this association proves to be causal, BV could be responsible for a substantial proportion of new HIV-1 infections in Africa. Normalization of vaginal flora in HIV-1-infected women could mitigate female-to-male HIV-1 transmission. TRIAL REGISTRATION: ClinicalTrials.com NCT00194519.

Development and validation of an in vivo Candida albicans biofilm denture model.

The most common form of oral candidiasis, denture-associated stomatitis, involves biofilm growth on an oral prosthetic surface. Cells in this unique environment are equipped to withstand host defenses and survive antifungal therapy. Studies of the biofilm process on dentures have primarily been limited to in vitro models. We developed a rodent acrylic denture model and characterized the Candida albicans and mixed oral bacterial flora biofilm formation, architecture, and drug resistance in vivo, using time course quantitative culture experiments, confocal microscopy, scanning electron microscopy, and antifungal susceptibility assays. We also examined the utility of the model for measurement of C. albicans gene expression and tested the impact of a specific gene product (Bcr1p) on biofilm formation. Finally, we assessed the mucosal host response to the denture biofilm and found the mucosal histopathology to be consistent with that of acute human denture stomatitis, demonstrating fungal invasion and neutrophil infiltration. This current oral denture model mimics human denture stomatitis and should be useful for testing the impact of gene disruption on biofilm formation, studying the impact of anti-infectives, examining the biology of mixed Candida-oral bacterial flora biofilm infections, and characterizing the host immunologic response to this disease process.

Efficacy of bilateral bronchoalveolar lavage for diagnosis of ventilator-associated pneumonia.

Ventilator-associated pneumonia (VAP) is a common nosocomial infection causing significant morbidity and mortality. The goal of this study was to determine the efficacy of bilateral versus unilateral bronchoalveolar lavage (BAL) for the detection of the causative bacterial agents of VAP. We retrospectively studied the quantitative bacterial cultures of 399 BAL sample pairs collected from 287 mechanically ventilated patients over a 5-year period at a U.S. tertiary-care teaching hospital. Trauma was the underlying illness in 69% of patients. No evidence of bacterial infection was found in 226 BAL pairs (56.6%). Among 173 positive BAL sample pairs, significant bacterial counts were detected exclusively in 6.4% of left-lung and 12.1% of right-lung samples. In contrast, 81.5% of positive sample pairs had significant bacterial counts in both lungs. All bacteria recovered at significant concentrations from bilateral samples would have been detected in a unilateral right-lung sample in 89% of positive sample pairs. Unilateral sampling would have failed to recover one or more significant isolates in 11% of positive pairs had only the right lung been sampled and in 16.7% had only the left lung been sampled. Our study shows that preferential sampling of the right lung improves the diagnostic efficacy of unilateral BAL for the detection of the etiologic agents of VAP. If bilateral sampling is performed, our results also indicate that pooling left- and right-lung samples for a single quantitative culture is comparable to processing samples individually.

Identification of plasmid-mediated AmpC beta-lactamases in Escherichia coli, Klebsiella spp., and proteus species can potentially improve reporting of cephalosporin susceptibility testing results.

The goal of this study was to determine if the interpretations of extended-spectrum and advanced-spectrum cephalosporins (ESCs and ASCs, respectively) for isolates of Enterobacteriaceae would be impacted by the results of aminophenylboronic acid (APBA) testing. Fifty-three isolates of Escherichia coli, 21 Klebsiella species, and 6 Proteus species that were resistant to at least one ESC were tested by disk diffusion with ceftazidime and cefotetan disks with and without APBA. Ceftazidime disks with and without clavulanic acid (CLAV) were also tested to confirm extended-spectrum beta-lactamase (ESBL) carriage. Twenty-nine (36.3%) isolates were only APBA test positive, 27 were only CLAV test positive, 2 were positive with both substrates, and 22 were negative with both substrates. Thirteen (41.9%) of the 31 APBA-test-positive isolates (all E. coli) tested susceptible to cefotaxime, ceftriaxone, or ceftazidime. Since clinical data suggest that AmpC-producing isolates should be reported as resistant to all ESCs, APBA testing can be helpful in identifying such organisms. Screening for AmpC-producing organisms using nonsusceptibility to cefoxitin and amoxicillin-clavulanate was less specific than APBA testing; it identified ESBL as well as AmpC-producing organisms. Only 18 of 31 APBA-positive isolates were positive by PCR for an AmpC beta-lactamase gene. Thus, testing with APBA could improve the accuracy of reporting ESCs, especially for E. coli. However, results of APBA and CLAV testing did not correlate well for isolates containing both AmpC beta-lactamases and ESBLs. Thus, additional data are needed before formal recommendations can be made on changing the reporting of ASC test results.

Bacteremic sepsis disturbs alveolar perfusion distribution in the lungs of rats.

OBJECTIVE: Sepsis often leads to lung injury, although the mechanisms that initiate this are unclear. One preinjury phenomenon that has not been explored previously is the effect of bacterial (nonlipopolysaccharide) sepsis on the distribution of alveolar perfusion. The goals of our studies were to measure this. DESIGN: Randomized, controlled, prospective animal study. SETTING: University animal laboratory. SUBJECTS: Male Sprague-Dawley rats (450-550 g). INTERVENTIONS: We induced sepsis by placing gelatin capsules containing Escherichia coli and Bacteroides fragilis into the abdomens of rats (n = 9). Empty capsules (n = 6) were placed into the abdomens of controls. After 24 hrs, 4-microm-diameter fluorescent latex particles (2 x 10(8)) were infused into the pulmonary circulation. Sepsis was induced in additional rats and controls to assess lung injury, as follows: Lung histology was performed on eight septic rats and on seven controls; lung lavage was performed on three septic rats and three controls after their plasma albumin had been labeled with Evans blue dye. MEASUREMENTS AND MAIN RESULTS: Confocal microscopy was used to prepare digital maps of latex particle trapping patterns (eight per lung). Analysis of these patterns revealed statistically more clustering (perfusion inhomogeneity) down to tissue volumes less than that of ten alveoli in septic lungs compared with controls (p < or = .05). Bacterial counts and neutrophil counts were significantly higher in the circulation of septic rats (p < or = .05). Blood pressures and arterial PO2s were unchanged. Cell counts in histological images were three-fold higher in septic lungs than in controls (p < or = .05). Lung lavage revealed 0.41 +/- 0.03 mL of plasma in the lungs of septic rats, and 0.06 +/- 0.05 mL in the lungs of controls (p < or = .05). CONCLUSIONS: Bacterial sepsis caused significant maldistribution of interalveolar perfusion in the lungs of rats in the absence of significant lung injury.

Vacuum-mixing significantly changes antibiotic elution characteristics of commercially available antibiotic-impregnated bone cements.

BACKGROUND: Evidence-based medicine indicates the use of antibiotic-impregnated polymethylmethacrylate bone cement during hip and knee replacement reduces the rate of prosthetic joint infection. In the United States, so-called off-label use of antibiotic-impregnated polymethylmethacrylate for primary joint replacement is increasing and multiple antibiotic-containing polymethylmethacrylate products are commercially available. However, there are sparse published data comparing the antibiotic elution characteristics of these bone cement products and the effect that vacuum-mixing has on antibiotic elution from these products. This study compares the antibiotic elution characteristics of six commercially available antibiotic polymethylmethacrylate formulations mixed under atmospheric pressure and vacuum conditions. METHODS: The antibiotic-impregnated polymethylmethacrylate products were mixed with use of a commonly employed intraoperative technique at atmospheric pressure and clinically relevant vacuum conditions. A standard Kirby-Bauer bioassay technique was subsequently used to quantify antibiotic elution from the products. An international infectious disease database was mined to determine antibiotic susceptibility of common bacteria causing prosthetic joint infection and to define the gentamicin concentration above which optimal antibiotic efficacy begins for these organisms. Statistical analyses incorporating the above susceptibility data were performed to compare antibiotic elution (1) among products mixed at atmospheric pressure, (2) among vacuum-mixed products, and (3) between atmospheric and vacuum-mixing for each individual product. RESULTS: Comparisons of antibiotic-loaded polymethylmethacrylate products mixed at atmospheric pressure indicated that significant antibiotic elution differences exist among the products. Comparisons of vacuum-mixed antibiotic-loaded polymethylmethacrylate products indicated that significant antibiotic elution differences exist among the products. When mixing under atmospheric pressure was compared with vacuum-mixing for each individual antibiotic polymethylmethacrylate product, vacuum-mixing significantly increased the clinically relevant cumulative antibiotic elution from three products but significantly decreased antibiotic elution from three other products. CONCLUSIONS: The method by which antibiotic-containing polymethylmethacrylate products are prepared significantly affects their antibiotic elution characteristics. The effect of vacuum-mixing on antibiotic elution is product-specific.

Bacteremia does not affect cellular uptake of ultrafine particles in the lungs of rats.

To assess the effects of intra-abdominal bacteremia on lung cellular function in vivo, we used electron microscopy to quantify the uptake of 6 nm diameter, albumin-coated colloidal gold particles (overall diam. 20.8 nm) by cells in the lungs of rats made septic by the introduction of live bacteria (E.coli and B. fragilis) into their abdomens. Gold particles were instilled into the trachea 24 hr after bacteremia induction, and lungs were harvested and prepared for electron microscopy 24 hr later. Because bacteremia produces an increase in metabolism, we hypothesized that this might be associated with increased cellular uptake of these particles and also with increased permeability of the alveolar epithelial barrier to them, as bacteremia is also associated with lung injury. We quantified particle uptake by counting particle densities (particles/µm²) within type I and type II epithelial cells, capillary endothelial cells, erythrocytes and neutrophils in the lungs of five septic rats and five sham-sepsis controls. We also counted particle densities within organelles of these cells (nuclei, mitochondria, type II cell lamellar bodies) and within the alveolar interstitium. We found particles to be present within all of these compartments, although we found no differences in particle densities between bacteremic rats and sham-sepsis controls. Our results suggest that these 6 nm particles were able to freely cross cell and organelle membranes, and further suggest that this ability was not altered by bacteremia.