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In men with nongonococcal urethritis (NGU), clinicians and patients rely on clinical cure to guide the need for additional testing/treatment and when to resume sex, respectively; however, discordant clinical and microbiological cure outcomes do occur. How accurately clinical cure reflects microbiological cure in specific sexually transmitted infections (STIs) is unclear.Men with NGU were tested for Neisseria gonorrhoeae, Chlamydia trachomatis (CT), Mycoplasma genitalium (MG), Trichomonas vaginalis, urethrotropic Neisseria meningitidis ST-11 clade strains, and Ureaplasma urealyticum (UU). Men received azithromycin 1 g and returned for a 1-month test-of-cure visit. In MG infections, we evaluated for the presence of macrolide resistance-mediating mutations (MRMs) and investigated alternate hypotheses for microbiological treatment failure using in situ shotgun metagenomic sequencing, phylogenetic analysis, multilocus sequence typing analyses, and quantitative PCR.Of 280 men with NGU, 121 were included in this analysis. In the monoinfection group, 52 had CT, 16 had MG, 7 had UU, 10 had mixed infection, and 36 men had idiopathic NGU. Clinical cure rates were 85% for CT, 100% for UU, 50% for MG, and 67% for idiopathic NGU. Clinical cure accurately predicted microbiological cure for all STIs, except MG. Discordant results were significantly associated with MG-NGU and predominantly reflected microbiological failure in men with clinical cure. Mycoplasma genitalium MRMs, but not MG load or strain, were strongly associated with microbiological failure.In azithromycin-treated NGU, clinical cure predicts microbiological cure for all STIs, except MG. Nongonococcal urethritis management should include MG testing and confirmation of microbiological cure in azithromycin-treated MG-NGU when MRM testing is unavailable.

Background In men with nongonococcal urethritis (NGU), clinicians and patients rely on the resolution of urethritis signs and symptoms to determine the need for additional testing/treatment and when to resume sexual activity, respectively. Whether clinical NGU cure correlates with microbiological clearance across sexually transmitted infections (STI) remains unknown. Methods Within the Idiopathic Urethritis Men’s Project (IUMP), we identified men with mono-infection NGU due to Chlamydia trachomatis, Mycoplasma genitalium, or Ureaplasma urealyticum. Men had presented to the Bell Flower Clinic in Indianapolis, Indiana, and were diagnosed with NGU and enrolled in IUMP. Men provided a urine specimen for STI testing by NAAT, received azithromycin 1gm and returned for a 1-month test-of-cure visit. At the test-of-cure visit, men were asked about interval antibiotic use, sexual activity, interval urethritis signs and symptoms, and repeat STI testing was performed. Clinical cure was defined as resolution of urethritis signs and symptoms. Microbiological cure was defined as clearance of STI at the test-of-cure visit. Significance was evaluated by Fisher’s exact test. Results Seventy-five men were included in this analysis, 52 with CT, 16 with MG, and 7 with UU. Clinical cure occurred in 50–100% and microbiological cure occurred in 31–100% of men. Discordant cures were more common in men with MG than CT (44% vs 15%, p = 0.102); no UU discordant cures occurred. Men with MG-NGU were significantly more likely to have a clinical cure with microbiological failure compared to CT-NGU (31% vs 4%, p = 0.0042). Clinical failure with microbiological cure occurred in approximately 10% of men. Conclusions Discordant NGU cure outcomes do occur and are more common in men with MG-NGU. The most common discordant cure outcome was clinical cure with microbiological failure. In men treated with azithromycin for NGU, MG testing should be included and a test-of-cure considered to ensure STI clearance.

Chlamydiae are obligate intracellular pathogens that rely on secreted effector proteins to establish their intracellular niche. In this issue of the Journal of Bacteriology , Yanatori et al. describe a screen for Chlamydia pneumoniae effectors, performed in Chlamydia trachomatis , which identified several new proteins that are translocated during infection (I).

Multiple species of obligate intracellular bacteria in the genus Chlamydia are important veterinary and/or human pathogens. These pathogens all share similar biphasic developmental cycles and transition between intracellular vegetative reticulate bodies and infectious elementary forms, but vary substantially in their host preferences and pathogenic potential. A lack of tools for genetic engineering of these organisms has long been an impediment to the study of their biology and pathogenesis. However, the refinement of approaches developed in C. trachomatis over the last 10 years, and adaptation of some of these approaches to other Chlamydia spp. in just the last few years, has opened exciting new possibilities for studying this ubiquitous group of important pathogens.

Cutaneous ulcers (CU) affect approximately 100,000 children in the tropics each year. While two-thirds of CU are caused by Treponema pallidum subspecies pertenue and Haemophilus ducreyi, the cause(s) of the remaining one-third is unknown., Exudative cutaneous ulcers (CU) in yaws-endemic areas are associated with Treponema pallidum subsp. pertenue (TP) and Haemophilus ducreyi (HD), but one-third of CU cases are idiopathic (IU). Using mass drug administration (MDA) of azithromycin, a yaws eradication campaign on Lihir Island in Papua New Guinea reduced but failed to eradicate yaws; IU rates remained constant throughout the campaign. To identify potential etiologies of IU, we obtained swabs of CU lesions (n = 279) and of the skin of asymptomatic controls (AC; n = 233) from the Lihir Island cohort and characterized their microbiomes using a metagenomics approach. CU bacterial communities were less diverse than those of the AC. Using real-time multiplex PCR with pathogen-specific primers, we separated CU specimens into HD-positive (HD+), TP+, HD+TP+, and IU groups. Each CU subgroup formed a distinct bacterial community, defined by the species detected and/or the relative abundances of species within each group. Streptococcus pyogenes was the most abundant organism in IU (22.65%) and was enriched in IU compared to other ulcer groups. Follow-up samples (n = 31) were obtained from nonhealed ulcers; the average relative abundance of S. pyogenes was 30.11% in not improved ulcers and 0.88% in improved ulcers, suggesting that S. pyogenes in the not improved ulcers may be azithromycin resistant. Catonella morbi was enriched in IU that lacked S. pyogenes. As some S. pyogenes and TP strains are macrolide resistant, penicillin may be the drug of choice for CU azithromycin treatment failures. Our study will aid in the design of diagnostic tests and selective therapies for CU.

Background Dysmenorrhea is highly prevalent; it places women at risk for other chronic pain conditions. There is a high degree of individual variability in menstrual pain severity, the number of painful sites, and co-occurring gastrointestinal symptoms. Distinct dysmenorrhea symptom-based phenotypes were previously identified, but the biological underpinnings of these phenotypes are less known. One underexplored contributor is the vaginal microbiome. The vaginal microbiota differs significantly among reproductive-age women and may modulate as well as amplify reproductive tract inflammation, which may contribute to dysmenorrhea symptoms. Objectives The objective of this study was to examine associations between dysmenorrhea symptom-based phenotypes and vaginal microbiome compositions on- and off-menses. Methods We conducted a prospective, longitudinal, pilot study of 20 women (aged 15-24) grouped into three dysmenorrhea symptom-based phenotypes: "mild localized pain," "severe localized pain," and "severe multiple pain and gastrointestinal symptoms." Over one menstrual cycle, participants provided vaginal swabs when they were on-menses and off-menses. We assayed the vaginal microbiome using 16S rRNA gene sequencing. Permutational multivariate analysis of variance tests were used to compare microbiome compositions across phenotypes, with heat maps generated to visualize the relative abundance of bacterial taxa. Results The vaginal microbiome compositions (n = 40) were different across the three phenotypes. After separating the on-menses (n = 20) and off-menses (n = 20) specimens, the statistically significant difference was seen on-menses, but not off-menses. Compared to the "mild localized pain" phenotype, participants in the "multiple severe symptoms" phenotype had a lower lactobacilli level and a higher abundance of Prevotella, Atopobium, and Gardnerella when on-menses. We also observed trends of differences across phenotypes in vaginal microbiome change from off- to on-menses. Discussion The study provides proof-of-concept data to support larger studies on associations between dysmenorrhea symptom-based phenotypes and vaginal microbiome that might lead to new intervention targets and/or biomarkers for dysmenorrhea. This line of research has the potential to inform precision dysmenorrhea treatment that can improve women's quality of life.

Chlamydia spp. productively infect mucosal epithelial cells of multiple anatomical sites, including the conjunctiva, lungs, gastrointestinal (GI) tract, and urogenital tract. We, and others, previously established that chlamydial GI tropism is mediated by distinct chromosomal and plasmid factors. In this study, we describe a genital infection-attenuated Chlamydia muridarum mutant (GIAM-1) that is profoundly and specifically attenuated in the murine genital tract. GIAM-1 infected the murine GI tract similarly to wild-type (WT) Chlamydia muridarum but did not productively infect the lower genital tract of female mice, ascend to infect the upper genital tract, or cause hydrosalpinx. However, GI infection of mice with GIAM-1 elicited a transmucosal immune response that protected against subsequent genital challenge with WT Chlamydia muridarum. Collectively, our results demonstrate that chlamydia mutants that are profoundly attenuated for specific organ tissues can be derived and demonstrate that live-attenuated vaccine strains that infect the GI tract, but do not elicit genital tract disease, could be used to protect against chlamydia genital tract infection and disease. IMPORTANCE Chlamydia is the most common sexually transmitted bacterial infection in the United States. Most chlamydia genital infections resolve without serious consequences; however, untreated infection in women can cause pelvic inflammatory disease and infertility. Antibiotics are very effective in treating chlamydia, but most genital infections in both men and women are asymptomatic and go undiagnosed. Therefore, there is a critical need for an effective vaccine. In this work, we show that a mutant chlamydia strain, having substantially reduced virulence for genital infection, colonizes the gastrointestinal tract and produces robust immunity to genital challenge with fully virulent wild-type chlamydia. These results are an important advance in understanding chlamydial virulence and provide compelling evidence that safe and effective live-attenuated chlamydia vaccines may be feasible.

Identifying pathogen-specific signs or symptoms of nongonococcal urethritis could improve syndromic management accuracy. We evaluated nongonococcal urethritis signs and symptoms in 220 men with single-pathogen infections (Chlamydia trachomatis, Mycoplasma genitalium, Trichomonas vaginalis, or Ureaplasma urealyticum) or idiopathic urethritis. No individual sign or symptom accurately predicted the infectious etiology.

Different variants of the obligate intracellular bacterial pathogen Chlamydia trachomatis cause the diseases trachoma and chlamydia. The trachoma strains cause chronic infections of the conjunctival epithelium and an intense inflammatory response that can lead to corneal damage, and trachoma is the most common cause of infectious blindness. The Chlamydia strains preferentially infect columnar epithelial cells of the cervix or urethra, and Chlamydia is the most common bacterial sexually transmitted infection. Chlamydia strains often elicit no symptoms but sometimes ascend the reproductive tract where they can elicit chronic inflammation and severe consequences, including pelvic inflammatory disease. Despite their distinct tissue tropisms, the genomes of the trachoma and Chlamydia strains are nearly identical (1). Chlamydia genital tropism has been linked to tryptophan synthase (TS), an enzyme that synthesizes tryptophan from indole (Fig. 1) (2, 3). Expression of the α-subunit (TrpA) and β-subunit (TrpB) of the TS holoenzyme is tightly regulated by an apo-repressor (TrpR) (4⇓⇓–7). In the presence of the corepressor tryptophan, TrpR binds an operator sequence ( trpO ) and blocks trpBA transcription. Low tryptophan levels promote TrpR release from trpO and allow TS expression. Chlamydia strains likely use TS to circumvent immune-regulated tryptophan catabolism, mediated by the host enzyme indoleamine 2,3′ dioxygenase (IDO1) (2). In contrast, most trachoma isolates do not express a functional TS (2). A study in PNAS by Sherchand and Aiyar (8 … [↵][1]1To whom correspondence may be addressed. Email: nelsonde{at}indiana.edu. [1]: #xref-corresp-1-1

IMPACT: This project seeks to identify unique host responses that are biomarkers for specific urethral pathogens, and which can be used in the development of point-of-care (POC) STI diagnostics. OBJECTIVES/GOALS: How Chlamydia trachomatis (CT) and other common STIs, e.g. Neisseria gonorrhoeae, evade immunity and elicit pathology in the male urethra is poorly understood. Our objective is to determine how STI-infected urethral epithelial cells, as well as the uninfected ‘bystander’ cells with which infected cells communicate, respond to CT and other STIs. METHODS/STUDY POPULATION: We evaluated how immortalized urethral cell lines - including transduced human urethral epithelial cells (THUECs) - respond to increasing doses of CT infectious particles using in vitro one-step progeny assays performed in the presence or absence of cycloheximide, a drug that inhibits eukaryotic protein synthesis. We will perform concurrent single-cell RNA sequencing (scRNA-seq) and multiplex cytokine analyses to determine how different CT doses impact the transcriptomes of infected and bystander urethral epithelial cells and modulate cytokine production of the overall monolayer. Results of these experiments will inform the feasibility of performing similar analyses in situ using urethral swabs from men with clinically diagnosed urethritis. RESULTS/ANTICIPATED RESULTS: Our results demonstrate that immune-competent urethral cell monolayers strongly resist CT infection, unless most of the cells are simultaneously infected. This suggests that uninfected bystander cells sense CT-infected cells and secrete soluble factors that may act to limit CT proliferation in infected cells and to inform remaining uninfected cells that a potential pathogen is present. We anticipate that our scRNA-seq and cytokine analyses will identify both specific effector pathways that protect against CT and intracellular signals that modulate them. We speculate that these pathways and signals may differ during infection with CT and other STIs. Importantly, we anticipate that our in vitro model of CT infection will be highly representative of in situ immune responses observed in urethras of infected men. DISCUSSION/SIGNIFICANCE OF FINDINGS: In men, common STIs including CT are usually managed syndromically due to a lack of POC diagnostics. By determining how STIs elicit urethral inflammation and identifying countermeasures that STIs use to evade urethral immunity, we can identify host responses that serve as biomarkers for urethritis, generally, and for specific urethral pathogens.

Neisseria meningitidis (Nm) clonal complex 11 (cc11) lineage is a hypervirulent pathogen responsible for outbreaks of invasive meningococcal disease, including among men who have sex with men, and is increasingly associated with urogenital infections. Recently, clusters of Nm urethritis have emerged primarily among heterosexual males in the United States. We determined that nonencapsulated meningococcal isolates from an ongoing Nm urethritis outbreak among epidemiologically unrelated men in Columbus, Ohio, are linked to increased Nm urethritis cases in multiple US cities, including Atlanta and Indianapolis, and that they form a unique clade (the US Nm urethritis clade, US_NmUC). The isolates belonged to the cc11 lineage 11.2/ET-15 with fine type of PorA P1.5–1, 10–8; FetA F3-6; PorB 2–2 and express a unique FHbp allele. A common molecular fingerprint of US_NmUC isolates was an IS1301 element in the intergenic region separating the capsule ctr-css operons and adjacent deletion of cssA/B/C and a part of csc , encoding the serogroup C capsule polymerase. This resulted in the loss of encapsulation and intrinsic lipooligosaccharide sialylation that may promote adherence to mucosal surfaces. Furthermore, we detected an IS1301-mediated inversion of an ∼20-kb sequence near the cps locus. Surprisingly, these isolates had acquired by gene conversion the complete gonococcal denitrification nor B- ani A gene cassette, and strains grow well anaerobically. The cc11 US_NmUC isolates causing urethritis clusters in the United States may have adapted to a urogenital environment by loss of capsule and gene conversion of the Neisseria gonorrheae norB - aniA cassette promoting anaerobic growth.

At a clinic in Indianapolis, Indiana, USA, we observed an increase in Neisseria gonorrhoeae-negative men with suspected gonococcal urethritis who had urethral cultures positive for N. meningitidis. We describe genomes of 2 of these N. meningitidis sequence type 11 complex urethritis isolates. Clinical evidence suggests these isolates may represent an emerging urethrotropic clade.

ObjectivesChlamydia trachomatis(CT) andMycoplasma genitalium(MG) cause the majority of non-gonococcal urethritis (NGU). The role ofUreaplasma urealyticum(UU) in NGU is unclear. Prior case–control studies that examined the association of UU and NGU may have been confounded by mixed infections and less stringent criteria for controls. The objective of this case–control study was to determine the prevalence and aetiology of mixed infections in men and assess if UU monoinfection is associated with NGU.MethodsWe identified 155 men with NGU and 103 controls. Behavioural and clinical information was obtained and men were tested forNeisseria gonorrhoeaeand CT, MG, UU andTrichomonas vaginalis(TV). Men who were five-pathogen negative were classified as idiopathic urethritis (IU).ResultsTwelve per cent of NGU cases in which a pathogen was identified had mixed infections, mostly UU coinfections with MG or CT; 27% had IU. In monoinfected NGU cases, 34% had CT, 17% had MG, 11% had UU and 2% had TV. In controls, pathogens were rarely identified, except for UU, which was present in 20%. Comparing cases and controls, NGU was associated with CT and MG monoinfections and mixed infections. UU monoinfection was not associated with NGU and was almost twice as prevalent in controls. Men in both the case and control groups who were younger and who reported no prior NGU diagnosis were more likely to have UU (OR 0.97 per year of age, 95% CI 0.94 to 0.998 and OR 6.3, 95% CI 1.4 to 28.5, respectively).ConclusionsMixed infections are common in men with NGU and most of these are UU coinfections with other pathogens that are well-established causes of NGU. UU monoinfections are not associated with NGU and are common in younger men and men who have never previously had NGU. Almost half of NGU cases are idiopathic.

Urethritis, or inflammation of the urethra, is one of the most common reasons men seek clinical care. Sexually transmitted pathogens including Neisseria gonorrhoeae are responsible for over half of the symptomatic urethritis cases in U.S. men. Recently, clinics in Indianapolis, Columbus, Atlanta, and other U.S. cities began to note increasing numbers of men presenting with urethritis and Gram-negative intracellular diplococci in their urethral smears who test negative for N. gonorrhoeae. Many of these discordant cases, which have periodically reached highs of more than 25% of presumed gonococcal cases in some sexually transmitted infection clinics in the U.S. Midwest, are infected with strains in a novel urethrotropic clade of Neisseria meningitidis ST-11 (US_NmUC). However, no cultivation-independent tests are available for the US_NmUC strains, and prior studies relied on microbial culture and genome sequencing to identify them. Here, we describe a PCR test that can identify the US_NmUC strains and distinguish them from commensal and invasive N. meningitidis strains as well as N. gonorrhoeae. Our SimpleProbe®-based real-time PCR assay targets a conserved nucleotide substitution in a horizontally acquired region of US_NmUC strain genomes. We applied the assay to 241 urine specimens whose microbial compositions had previously been determined by deep shotgun metagenomic sequencing. The assay detected the single US_NmUC positive case in this cohort, with no false positives. Overall, our simple and readily adaptable assay could facilitate investigation of the pathogenesis and epidemiology of the US_NmUC clade.

Background Persistent NGU occurs when symptoms persist after empiric NGU treatment and has been associated with Mycoplasma genitalium (MG) infection. The prevalence and etiology of persistent NGU in men remains largely unknown. Methods Within the Idiopathic Urethritis Men’s Project cohort study, we recruited men with NGU. NGU was diagnosed by the presence of urethritis signs and/or symptoms and urethral Gram stain with ≥5 PMN/HPF. Men were treated with 1 gm azithromycin and returned for a 1-month test-of-cure visit. At the test-of-cure visit, men were asked about post-treatment symptom outcomes and partner treatment. A first-catch urine specimen was obtained at both visits for five-pathogen testing for Neisseria gonorrhoeae (NG),Chlamydia trachomatis (CT), MG, Trichomonas vaginalis (TV), and Ureaplasma urealyticum (UU). NG-positive cases were excluded and five-pathogen-negative cases were classified as idiopathic urethritis (IU). Post-treatment symptom outcomes were: (1) resolved, (2) resolved then recurred, or (3) persisted unchanged. Results One hundred twenty-four men are included in this study. The median age was 28, 52% were African American, and 86% self-identified as heterosexual. All men reported urethral symptoms and 98% had a discharge on exam at baseline. Symptoms resolved completely in 91 (73%) men. Symptoms resolved then recurred or persisted unchanged in 12 (10%) and 21 (17%) men, respectively. Excluding men with untreated partners (N = 9, 28%), a different pathogen was identified in 5 (50%) and 4 (25%) men with recurrent and persistent symptoms, respectively. In men with the same pathogen identified (N = 15), 53% were IU, 33% were MG, 7% were CT, and 7% were UU. Conclusion Persistent NGU occurs in approximately 25% of azithromycin-treated men and is related to a new infection in up to 50% of cases. In men with persistent symptoms and the same infection identified at the test-of-cure visit, MG and IU comprised 86% of cases, which suggests that MG and IU-associated organisms may be resistant to azithromycin. Disclosure No significant relationships.

Background Syndromic management remains the standard nongonococcal urethritis (NGU) treatment approach. Whether pathogen-specific signs/symptoms inform treatment decisions remains unclear. We identified men with single- and mixed-pathogen NGU and assessed for the presence of pathogen-specific signs or symptoms to improve syndromic management. Methods As part of an ongoing cohort study (the Idiopathic Urethritis Men’s Project [IUMP]), we recruited men with NGU. NGU was diagnosed by signs and/or symptoms of urethritis, and a urethral Gram stain with ≥5 neutrophils per high-power field without evidence of gram negative intracellular diplococci. Participants underwent a clinical history and physical exam, which documented specific self-reported symptoms and clinician observed signs. Single- and mixed-infections were identified by NAAT testing of first-catch urine for Neisseria gonorrhoeae (NG), Chlamydia trachomatis (CT), Mycoplasma genitalium (MG), Trichomonas vaginalis (TV), and Ureaplasma urealyticum (UU); five-pathogen-negative cases were classified as idiopathic urethritis (IU). Results One hundred fifty-five men with NGU are included in this analysis. The median age was 28 (range 18–63), 101 (65%) were African American, and 135 (87%) self-identified as heterosexual. The most commonly reported symptom was urethral discharge (92%), followed by burning/tingling (37%), and dysuria (28%). Over half of these men reported more than one symptom (58%). Single-pathogen NGU was detected in 99 (64%) men, mixed-pathogen in 14 (9%), and IU in 42 (27%). For single pathogen NGU, 53 (34%) had CT, 26 (17%) had MG, 3 (2%) had TV, and 17 (11%) had UU. We compared single-pathogen NGU, mixed-infection and IU for differences in signs or symptoms and found no pathogen-specific differences. Conclusion In men with NGU, no pathogen-specific signs and symptoms were identified that could inform treatment decisions. Pathogen-specific point-of-care tests are needed. Disclosure No significant relationships.

Background The ability to quantify the organism load of Chlamydia trachomatis (CT) using a commercial assay could expand insights from epidemiological studies. This approach can be applied to routine diagnostic testing, and multiple specimen types. Approximate CT organism load was determined in urine from men with NGU, with and without co-infections, by comparing the results from each positive sample to a set of CT standards using the Abbott Realtime m2000 (m2000) platform. Methods Urine specimens, collected from men participating in the Idiopathic Urethritis Men’s Project (IUMP), were tested on the m2000 for CT. Additional testing included Neisseria gonorrhoeae, Mycoplasma genitalium, Trichomonas vaginalis, and Ureaplasma urealyticum. Standards were prepared by diluting CT elementary bodies (EB) into the collection device at six concentrations. CT organism load was determined by comparing the instrument generated delta cycle (DC) value from each CT positive urine to the standard curve. Calculated means were compared by t-test (p Results Two hundred and six men were tested for CT and 83 (40.3%) were positive. The DC values for 81/83 (97.3%) CT positive samples fell within the range of the standard curve. The mean DC value was 12.15 (range 0.19–16.96) which equated to a mean CT organism load of 1.4×106 EB/ml urine (range 2.22×102-9.97×106). There was no difference between the mean organism load in specimens from men who did and did not have co-infections with other STIs, 2.04×106 versus 1.38×106 EB/ml, (p≥0.05). Conclusion CT load determination can be performed on urine specimens using the m2000. This could facilitate straightforward load determination in settings where routine testing is performed. In men with NGU, the CT organism load is high and no difference in CT load was observed in men with CT mono-infections and men co-infected with CT and other STIs. Disclosure No significant relationships.

BACKGROUND. Rectal infection with Chlamydia trachomatis (CT) is frequent in women who deny receptive anal sex and is thought to arise from autoinoculation of the rectum from vaginal secretions. An alternate hypothesis is that oral sex inoculates and establishes gastrointestinal tract infection. Distinguishing these hypotheses is difficult in women. In men, autoinoculation is unlikely and heterosexual men frequently perform oral sex, but rarely participate in receptive anal exposure behaviors. METHODS. We enrolled high-risk men with and without nongonococcal urethritis (NGU) who presented to a sexually transmitted infection clinic in Indianapolis, Indiana. Urine and rectal swabs were collected and tested for urogenital and rectal CT, Neisseria gonorrhoeae (NG), and Mycoplasma genitalium (MG). Men completed surveys concerning symptoms, sexual orientation, and detailed recent and lifetime oral and anal sexual behaviors. RESULTS. Rectal CT was detected in 2/84 (2.4%) heterosexual men who reported cunnilingus, but no lifetime receptive anal behaviors. All of the men who denied receptive anal behaviors were negative for rectal NG and MG. In homosexual and bisexual men, rectal CT prevalence was high (9.7%) and rectal NG (4.8%) and MG (4.8%) were also detected. CONCLUSIONS. We detected rectal CT infections in heterosexual men who reported cunnilingus but denied receptive anal behaviors. Oral sex may be a risk factor for rectal CT infection via oral inoculation of the gastrointestinal tract. SUMMARY. We observed a low incidence of rectal C. trachomatis infection in heterosexual men.

Multiple obligatory intracellular bacteria in the genus Chlamydia are important pathogens. In humans, strains of C. trachomatis cause trachoma, chlamydia, and lymphogranuloma venereum. These diseases are all associated with extended courses of infection and reinfection that likely reflect the ability of chlamydiae to evade various aspects of host immune responses. Interferon-stimulated genes, driven in part by the cytokine interferon gamma, restrict the host range of various Chlamydia species, but how these pathogens evade interferon-stimulated genes in their definitive host is poorly understood. Various Chlamydia species can inhibit death of their host cells and may have evolved this strategy to evade prodeath signals elicited by host immune responses. We present evidence that chlamydia-induced programmed cell death resistance evolved to counter interferon- and immune-mediated killing of Chlamydia-infected cells., Interferon-regulated immune defenses protect mammals from pathogenically diverse obligate intracellular bacterial pathogens of the genus Chlamydia. Interferon gamma (IFN-γ) is especially important in controlling the virulence of Chlamydia species and thus impacts the modeling of human chlamydial infection and disease in mice. How IFN-γ contributes to cell-autonomous defenses against Chlamydia species and how these pathogens evade IFN-γ-mediated immunity in their natural hosts are not well understood. We conducted a genetic screen which identified 31 IFN-γ-sensitive (Igs) mutants of the mouse model pathogen Chlamydia muridarum. Genetic suppressor analysis and lateral gene transfer were used to map the phenotype of one of these mutants, Igs4, to a missense mutation in a putative chlamydial inclusion membrane protein, TC0574. We observed the lytic destruction of Igs4-occupied inclusions and accompanying host cell death in response to IFN-γ priming or various proapoptotic stimuli. However, Igs4 was insensitive to IFN-γ-regulated cell-autonomous defenses previously implicated in anti-Chlamydia trachomatis host defense in mice. Igs4 inclusion integrity was restored by caspase inhibitors, indicating that the IFN-γ-mediated destruction of Igs4 inclusions is dependent upon the function of caspases or related prodeath cysteine proteases. We further demonstrated that the Igs4 mutant is immune restricted in an IFN-γ-dependent manner in a mouse infection model, thereby implicating IFN-γ-mediated inclusion destruction and host cell death as potent in vivo host defense mechanisms to which wild-type C. muridarum is resistant. Overall, our results suggest that C. muridarum evolved resistance mechanisms to counter IFN-γ-elicited programmed cell death and the associated destruction of intravacuolar pathogens.

We compared the gut microbial populations in 100 women, from rural Ghana and urban US [50% lean (BMI 2) and 50% obese (BMI ≥ 30 kg/m2)] to examine the ecological co-occurrence network topology of the gut microbiota as well as the relationship of short chain fatty acids (SCFAs) with obesity. Ghanaians consumed significantly more dietary fiber, had greater microbial alpha-diversity, different beta-diversity, and had a greater concentration of total fecal SCFAs (p-value Bacteroides uniformis was significantly more abundant in lean women, irrespective of country (FDR corrected p Ruminococcus callidus, Prevotella copri, and Escherichia coli, and smaller proportions of Lachnospiraceae, Bacteroides and Parabacteroides. Lean Ghanaians had a significantly greater abundance of predicted microbial genes that catalyzed the production of butyric acid via the fermentation of pyruvate or branched amino-acids, while obese Ghanaians and US women (irrespective of BMI) had a significantly greater abundance of predicted microbial genes that encoded for enzymes associated with the fermentation of amino-acids such as alanine, aspartate, lysine and glutamate. Similar to lean Ghanaian women, mice humanized with stool from the lean Ghanaian participant had a significantly lower abundance of family Lachnospiraceae and genus Bacteroides and Parabacteroides, and were resistant to obesity following 6-weeks of high fat feeding (p-value Ffa) receptor Ffa2, in spite of similar fecal SCFAs concentrations. We demonstrate that the association between obesity resistance and increased predicted ecological connectivity and stability of the lean Ghanaian microbiota, as well as increased local SCFA receptor level, provides evidence of the importance of robust gut ecologic network in obesity.

The cryptic plasmid is essential for Chlamydia muridarum dissemination from the genital tract to the gastrointestinal (GI) tract. Following intravaginal inoculation, a C. muridarum strain deficient in plasmid-encoded pGP3 or pGP4 but not pGP5, pGP7, or pGP8 failed to spread to the mouse gastrointestinal tract, although mice infected with these strains developed productive genital tract infections. pGP3- or pGP4-deficient strains also failed to colonize the gastrointestinal tract when delivered intragastrically. pGP4 regulates pGP3, while pGP3 does not affect pGP4 expression, indicating that pGP3 is critical for C. muridarum colonization of the gastrointestinal tract. Mutants deficient in GlgA, a chromosome-encoded protein regulated by pGP4, also consistently colonized the mouse gastrointestinal tract. Interestingly, C. muridarum colonization of the gastrointestinal tract positively correlated with pathogenicity in the upper genital tract. pGP3-deficient C. muridarum strains did not induce hydrosalpinx or spread to the GI tract even when delivered to the oviduct by intrabursal inoculation. Thus, the current study not only has revealed that pGP3 is a novel chlamydial colonization factor in the gastrointestinal tract but also has laid a foundation for investigating the significance of gastrointestinal Chlamydia .

Pathogenically diverse Chlamydia spp. can have surprisingly similar genomes. Chlamydia trachomatis isolates that cause trachoma, sexually transmitted genital tract infections (chlamydia), and invasive lymphogranuloma venereum (LGV) and the murine strain Chlamydia muridarum share 99% of their gene content. A region of high genomic diversity between Chlamydia spp. termed the plasticity zone (PZ) may encode niche-specific virulence determinants that dictate pathogenic diversity. We hypothesized that PZ genes might mediate the greater virulence and gamma interferon (IFN-γ) resistance of C. muridarum compared to C. trachomatis in the murine genital tract. To test this hypothesis, we isolated and characterized a series of C. muridarum PZ nonsense mutants. Strains with nonsense mutations in chlamydial cytotoxins, guaBA-add , and a phospholipase D homolog developed normally in cell culture. Two of the cytotoxin mutants were less cytotoxic than the wild type, suggesting that the cytotoxins may be functional. However, none of the PZ nonsense mutants exhibited increased IFN-γ sensitivity in cell culture or were profoundly attenuated in a murine genital tract infection model. Our results suggest that C. muridarum PZ genes are transcribed—and some may produce functional proteins—but are dispensable for infection of the murine genital tract.

Objectives Characterization of urinary bacterial microbiome and antimicrobial peptides after burn injury to identify potential mechanisms leading to urinary tract infections and associated morbidities in burn patients. Design Retrospective cohort study using human urine from control and burn subjects. Setting University research laboratory. Patients Burn patients. Interventions None. Measurements and main results Urine samples from catheterized burn patients were collected hourly for up to 40 hours. Control urine was collected from "healthy" volunteers. The urinary bacterial microbiome and antimicrobial peptide levels and activity were compared with patient outcomes. We observed a significant increase in urinary microbial diversity in burn patients versus controls, which positively correlated with a larger percent burn and with the development of urinary tract infection and sepsis postadmission, regardless of age or gender. Urinary psoriasin and β-defensin antimicrobial peptide levels were significantly reduced in burn patients at 1 and 40 hours postadmission. We observed a shift in antimicrobial peptide hydrophobicity and activity between control and burn patients when urinary fractions were tested against Escherichia coli and Enterococcus faecalis urinary tract infection isolates. Furthermore, the antimicrobial peptide activity in burn patients was more effective against E. coli than E. faecalis. Urinary tract infection-positive burn patients with altered urinary antimicrobial peptide activity developed either an E. faecalis or Pseudomonas aeruginosa urinary tract infection, suggesting a role for urinary antimicrobial peptides in susceptibility to select uropathogens. Conclusions Our data reveal potential links for urinary tract infection development and several morbidities in burn patients through alterations in the urinary microbiome and antimicrobial peptides. Overall, this study supports the concept that early assessment of urinary antimicrobial peptide responses and the bacterial microbiome may be used to predict susceptibility to urinary tract infections and sepsis in burn patients.

The cytokine gamma interferon (IFN-γ) induces cell-autonomous immunity to combat infections with intracellular pathogens, such as the bacterium Chlamydia trachomatis. The present study demonstrates that IFN-γ-primed human cells ubiquitinate and eliminate intracellular Chlamydia-containing vacuoles, so-called inclusions. We previously described how IFN-γ-inducible immunity-related GTPases (IRGs) employ ubiquitin systems to mark inclusions for destruction in mouse cells and, furthermore, showed that the rodent pathogen Chlamydia muridarum blocks ubiquitination of its inclusions by interfering with mouse IRG function. Here, we report that ubiquitination of inclusions in human cells is independent of IRG and thus distinct from the murine pathway. We show that C. muridarum is susceptible to inclusion ubiquitination in human cells, while the closely related human pathogen C. trachomatis is resistant. C. muridarum, but not C. trachomatis, inclusions attract several markers of cell-autonomous immunity, including the ubiquitin-binding protein p62, the ubiquitin-like protein LC3, and guanylate-binding protein 1. Consequently, we find that IFN-γ priming of human epithelial cells triggers the elimination of C. muridarum, but not C. trachomatis, inclusions. This newly described defense pathway is independent of indole-2,3-dioxygenase, a known IFN-γ-inducible anti-Chlamydia resistance factor. Collectively, our observations indicate that C. trachomatis evolved mechanisms to avoid a human-specific, ubiquitin-mediated response as part of its unique adaptation to its human host., IMPORTANCE Chlamydia trachomatis is the leading cause of sexually transmitted bacterial infections and responsible for significant morbidity, including pelvic inflammatory disease, infertility, and ectopic pregnancies in women. As an obligate intracellular pathogen, C. trachomatis is in perpetual conflict with cell-intrinsic defense programs executed by its human host. Our study defines a novel anti-Chlamydia host resistance pathway active in human epithelial cells. This defense program promotes the deposition of the small antimicrobial protein ubiquitin on vacuoles containing Chlamydia. We show that this ubiquitin-based resistance pathway of human cells is highly effective against a Chlamydia species adapted to rodents but ineffective against human-adapted C. trachomatis. This observation indicates that C. trachomatis evolved strategies to avoid entrapment within ubiquitin-labeled vacuoles as part of its adaptation to the human innate immune system.

OBJECTIVES/SPECIFIC AIMS: Objectives and goals of this study will be to: (1) compare fecal microbiota and fecal organic acids in irritable bowel syndrome (IBS) patients and controls and (2) investigate the association between colonic transit and fecal microbiota in IBS patients and controls. METHODS/STUDY POPULATION: We propose an investigation of fecal organic acids, colonic transit and fecal microbiota in 36 IBS patients and 18 healthy controls. The target population will be adults ages 18–65 years meeting Rome IV criteria for IBS (both diarrhea- and constipation-predominant, IBS-D and IBS-C) and asymptomatic controls. Exclusion criteria are: (a) history of microscopic colitis, inflammatory bowel disease, celiac disease, visceral cancer, chronic infectious disease, immunodeficiency, uncontrolled thyroid disease, liver disease, or elevated AST/ALT>2.0× the upper limit of normal, (b) prior radiation therapy of the abdomen or abdominal surgeries with the exception of appendectomy or cholecystectomy >6 months before study initiation, (c) ingestion of prescription, over the counter, or herbal medications affecting gastrointestinal transit or study interpretation within 6 months of study initiation for controls or within 2 days before study initiation for IBS patients, (d) pregnant females, (e) antibiotic usage within 3 months before study participation, (f) prebiotic or probiotic usage within the 2 weeks before study initiation, (g) tobacco users. Primary outcomes will be fecal bile acid excretion and profile, short-chain fatty acid excretion and profile, colonic transit, and fecal microbiota. Secondary outcomes will be stool characteristics based on responses to validated bowel diaries. Stool samples will be collected from participants during the last 2 days of a 4-day 100 g fat diet and split into 3 samples for fecal microbiota, SCFA, and bile acid analysis and frozen. Frozen aliquots will be shipped to the Metabolite Profiling Facility at Purdue University and the Mayo Clinic Department of Laboratory Medicine and Pathology for SCFA and bile acid measurements, respectively. Analysis of fecal microbiota will be performed in the research laboratory of Dr David Nelson in collaboration with bioinformatics expertise affiliated with the Nelson lab. Colonic transit time will be measured with the previously validated method using radio-opaque markers. Generalized linear models will be used as the analysis framework for comparing study endpoints among groups. RESULTS/ANTICIPATED RESULTS: This study seeks to examine the innovative concept that specific microbial signatures are associated with increased fecal excretion of organic acids to provide unique insights on a potential mechanistic link between altered intraluminal organic acids and fecal microbiota. DISCUSSION/SIGNIFICANCE OF IMPACT: Results may lead to development of targets for novel therapies and diagnostic biomarkers for IBS, emphasizing the role of the fecal metabolome.

Small intestinal bacterial overgrowth (SIBO) has historically been described in the context of maldigestion and malabsorption in patients with anatomic abnormalities (e.g., fistulae, strictures, bowel reconstruction, bowel obstruction), underlying dysmotility secondary to diabetes or scleroderma, and hypochlorhydria (1). This syndrome has also been implicated in the pathophysiology of numerous gastrointestinal and hepatobiliary diseases including common conditions such as irritable bowel syndrome (IBS) (2) and functional dyspepsia (3). Typical SIBO-associated symptoms include bloating, diarrhea, abdominal pain, and gas. SIBO symptoms may overlap with those of functional gastrointestinal disorders. However, the role of SIBO in the pathogenesis of functional gastrointestinal disorders remains controversial given that its definition relies on tests, including breath test and culture of small bowel aspirates, which have not been appropriately validated (4). Indirect techniques such as breath testing have important limitations related to heterogeneous testing methods and lack of standardization in interpretation. To address these limitations, guidelines for breath testing indications, performance, and interpretation were recently published in a North American consensus document (5). Yet, there remain important knowledge gaps in how these tests reflect the microbial landscape of the human small intestine. Small bowel aspirates and culture were previously considered the gold standard for the diagnosis of SIBO. SIBO has traditionally been defined by abnormally high amounts of bacteria in the small intestine and more precisely by >105 colony-forming units (CFU)/mL of colonic-type bacteria from jejunal aspirates (6). However, the validity of this approach has been questioned as the proposed cutoff of 105 seemed more predictive of postsurgical patients in a previous systematic review (7). Others have suggested that lower cutoff values of 103 CFU/mL be considered as diagnostic of SIBO (5), especially if coliforms are present (8). Standard culture-based techniques are limited in their ability to characterize fully the complex diversity of the small intestinal microbiome, particularly as a large proportion of bacterial species remain uncultured (9). In light of increasing evidence supporting the hypothesis that the intestinal microbiome serves a key role in host physiology and gastrointestinal disease, better characterization of the small intestinal microbiome will be a critical step in developing novel diagnostic and therapeutic strategies in a number of human diseases and disorders. In the recent years, the introduction of culture-independent techniques has dramatically expanded our ability to characterize the complexity of the intestinal microbiota. It also has increased our knowledge of the culture conditions needed to detect and identify previously difficult to culture organisms (10). Integration of rapid molecular approaches (11) for the assessment of SIBO in conditions such as IBS has yielded promising data including evidence for decreased microbial diversity from duodenal aspirates in patients with IBS compared with control subjects (12). However, these findings are preliminary. Overall, there are limited data reporting detailed characterizations of the human small intestinal microbiome. It may be hypothesized that alterations in the small intestinal microbiome are associated with culture-proven SIBO, which in turn could suggest that although imperfect, culture-proven SIBO is at least a useful surrogate marker of small intestinal dysbiosis. However, further investigation is needed to (i) explore the role of microbial diversity in SIBO using culture-independent approaches and (ii) describe the interactions between the host and small intestinal microbiome in the context of gastrointestinal disease. This study characterizes small intestinal microbial composition of small bowel aspirates and mucosal specimens in patients with and without a diagnosis of SIBO defined by qualitative and quantitative culture of proximal small bowel aspirates.

Objectives. Our goal was to provide current estimates of alcohol-attributable cancer mortality and years of potential life lost (YPLL) in the United States. Methods. We used 2 methods to calculate population-attributable fractions. We based relative risks on meta-analyses published since 2000, and adult alcohol consumption on data from the 2009 Alcohol Epidemiologic Data System, 2009 Behavioral Risk Factor Surveillance System, and 2009–2010 National Alcohol Survey. Results. Alcohol consumption resulted in an estimated 18 200 to 21 300 cancer deaths, or 3.2% to 3.7% of all US cancer deaths. The majority of alcohol-attributable female cancer deaths were from breast cancer (56% to 66%), whereas upper airway and esophageal cancer deaths were more common among men (53% to 71%). Alcohol-attributable cancers resulted in 17.0 to 19.1 YPLL for each death. Daily consumption of up to 20 grams of alcohol (≤ 1.5 drinks) accounted for 26% to 35% of alcohol-attributable cancer deaths. Conclusions. Alcohol remains a major contributor to cancer mortality and YPLL. Higher consumption increases risk but there is no safe threshold for alcohol and cancer risk. Reducing alcohol consumption is an important and underemphasized cancer prevention strategy.

Compelling evidence indicates that two autosomal recessive Parkinson's disease genes, PINK1 (PARK6) and Parkin (PARK2), cooperate to mediate the autophagic clearance of damaged mitochondria (mitophagy). Mutations in the F-box domain-containing protein Fbxo7 (encoded by PARK15) also cause early-onset autosomal recessive Parkinson's disease, by an unknown mechanism. Here we show that Fbxo7 participates in mitochondrial maintenance through direct interaction with PINK1 and Parkin and acts in Parkin-mediated mitophagy. Cells with reduced Fbxo7 expression showed deficiencies in translocation of Parkin to mitochondria, ubiquitination of mitofusin 1 and mitophagy. In Drosophila, ectopic overexpression of Fbxo7 rescued loss of Parkin, supporting a functional relationship between the two proteins. Parkinson's disease-causing mutations in Fbxo7 interfered with this process, emphasizing the importance of mitochondrial dysfunction in Parkinson's disease pathogenesis. © 2013 Nature America, Inc. All rights reserved.

In brief The authors surveyed equestrians to determine an injury profile. Based on responses from 2,195 frequent riders, the study confirms previous findings regarding injury rates, injury sites, and helmet use. New findings include a large number of neck and back injuries and a likelihood of injured riders to be between 15 and 44 years of age, ride English style, and have less than 10 years of riding experience. Data also show that many horseback-riding injuries are treated in physicians' offices. The high percentage of injured patients who suffered prolonged disability underscores the need for physicians to counsel horseback-riding patients about hazards and safety measures-especially helmet use.

Traditionally, the male urethra in healthy men was believed to be sterile or only transiently colonized by microorganisms. This view reflected technical limitations in studying this site that were not easily surmounted until methods for cultivation-independent bacterial identification became available. First, while the obvious analogous compartment to the male urethra may be the female urinary tract, given the function of the penis, the male urethral compartment is most often compared to the vaginal microenvironment. A simple Gram stain of these two body sites demonstrates substantial differences. Vaginal samples from healthy women typically contain significant numbers of epithelial cells and a large number of microorganisms including Lactobacillus and Candida species among others (see chapter 12). In contrast, it is unusual to identify similar numbers of microorganisms by simple Gram stain of urethral smears obtained from healthy men. An unintended consequence has been that far less effort has gone into characterizing the relatively sparse microorganisms that are routinely observed, and ignored, in urethral specimens from healthy men. As described later in this chapter, recent cultivation-independent surveys of male urethral swab, urine, and semen specimens have confirmed that the male urethra harbors smaller absolute numbers of microorganisms than the vagina but have also identified a surprisingly diverse array of bacterial taxa, many of which correspond to pathogens and commensal organisms found in the vagina. This has led to the reemerging idea that the microbiology of the male urethra may be more complex than is appreciated and that the composition of urethral microbial communities, collectively the urethral microbiome, might play a role in male urethral health and disease analogous to the role of the vaginal microbiome.

Aims Assess long-term trends of the correlation between alcohol sales data and survey data. Design Analyses of state alcohol consumption data from the US Alcohol Epidemiologic Data System based on sales, tax receipts or alcohol shipments. Cross-sectional, state annual estimates of alcohol-related measures for adults from the US Behavioral Risk Factor Surveillance System using telephone surveys. Setting United States. Participants State alcohol tax authorities, alcohol vendors, alcohol industry (sales data) and randomly selected adults aged >/= 18 years 1993-2006 (survey data). Measurements State-level per capita annual alcohol consumption estimates from sales data. Self-reported alcohol consumption, current drinking, heavy drinking, binge drinking and alcohol-impaired driving from surveys. Correlation coefficients were calculated using linear regression models. Findings State survey estimates of consumption accounted for a median of 22% to 32% of state sales data across years. Nevertheless, state consumption estimates from both sources were strongly correlated with annual r-values ranging from 0.55-0.71. State sales data had moderate-to-strong correlations with survey estimates of current drinking, heavy drinking and binge drinking (range of r-values across years: 0.57-0.65; 0.33-0.70 and 0.45-0.61, respectively), but a weaker correlation with alcohol-impaired driving (range of r-values: 0.24-0.56). There were no trends in the magnitude of correlation coefficients. Conclusions Although state surveys substantially underestimated alcohol consumption, the consistency of the strength of the association between sales consumption and survey data for most alcohol measures suggest both data sources continue to provide valuable information. These findings support and extend the distribution of consumption model and single distribution theory, suggesting that both sales and survey data are useful for monitoring population changes in alcohol use. Language: en

For biologic and behavioral reasons, cigarette smokers weigh less than nonsmokers. Thus, cigarette smoking may modify the association between media use and obesity. The authors examined whether the association between media use and overweight and obesity was modified by cigarette smoking by analyzing 8,467 adults (> or =20 years) from the 1999-2002 National Health and Nutrition Examination Survey. Odds ratios and 95% confidence intervals of overweight and obesity were estimated by use of multinomial regression. To examine effect modification, the authors created separate regression models for smokers and nonsmokers (p(interaction) = 0.002). Nonsmokers using media 4 or more hours daily were 3.9 times more likely to be obese (95% confidence interval (CI): 2.9, 5.2) and 1.6 times more likely to be overweight (95% CI: 1.3, 2.0) compared with those reporting less than 1 hour/day of media use. Among smokers, media use 4 or more hours daily was not significantly associated with increased odds of obesity (odds ratio = 1.3, 95% CI: 0.8, 2.2) or overweight (odds ratio = 1.4, 95% CI: 1.0, 1.9). Media use was associated with overweight and obesity and modified by cigarette smoking. Cigarette smoking should be evaluated as an effect modifier in studies of media use and obesity.

The purpose of this study was to examine the career paths of alumni from the National Cancer Institute (NCI) Cancer Prevention Fellowship Program (CPFP), a structured in-house postdoctoral training program of 3-4 years duration, and specifically what proportion of the alumni were currently performing cancer prevention-related activities. The analyses here included 119 CPFP alumni and 85 unsuccessful CPFP applicants, all of whom completed postdoctoral training between 1987-2011 and are currently employed. Postdoctoral training experiences and current career outcomes data were collected via online surveys. Differences between groups were assessed using chi-square and Fisher's exact test p-values and subsequent regression analyses adjusted for differences between the groups. Compared to 15.3% of unsuccessful CPFP applicants, 52.1% of CPFP alumni (odds ratio [OR] = 4.99, 95% confidence interval [95% CI): 1.91-13.0) were currently spending the majority of their time working in cancer prevention. Among those doing any cancer prevention-focused work, 54.3% of CPFP alumni spent the majority of their time performing cancer prevention research activities when compared to 25.5% of unsuccessful applicants (OR = 4.26, 95% CI: 1.38-13.2). In addition to the independent effect of the NCI CPFP, scientific discipline, and employment sector were also associated with currently working in cancer prevention and involvement in cancer prevention research-related activities. These results from a structured postdoctoral training program are relevant not only to the cancer prevention community but also to those interested in evaluating alignment of postdoctoral training programs with available and desired career paths more broadly.

Two patients with Bruton's X-linked agammaglobulinemia are described with bacteremia and skin/bone infection due to an organism which by 16S rRNA gene sequence analysis was most closely related to "Flexispira" rappini (and thus designated a Flexispira-like organism, FLO) and more distantly related to the Helicobacter species. The organism required microaerobic conditions and, supplemental H(2) gas for growth and was reliably stained with acridine orange. In common with Helicobacter cinaedi infections, the focus of the FLO infection was in one case in the blood vessels or lymphatics of an extremity and in the other case in the skin and adjacent bone of an extremity. In both cases, prolonged IV antibiotic therapy was necessary to clear the infection. The susceptibility of XLA patients to FLO infection appears to be related to the fact that XLA is associated with severe B cell (humoral) immunodeficiency and thus these patients have difficulty with intravascular or intralymphatic infection. These findings elucidate the nature of FLO infections in humans and point the way to their detection and treatment.

OBJECTIVE: To examine trends in diabetes prevalence in the U.S. RESEARCH DESIGN AND METHODS: This study was conducted via telephone surveys in states that participated in the Behavioral Risk Factor Surveillance System between 1990 and 1998. The participants consisted of noninstitutionalized adults aged 18 years or older. The main outcome measure was self-reported diabetes. RESULTS: The prevalence of diabetes rose from 4.9% in 1990 to 6.5% in 1998--an increase of 33%. Increases were observed in both sexes, all ages, all ethnic groups, all education levels, and nearly all states. Changes in prevalence varied by state. The prevalence of diabetes was highly correlated with the prevalence of obesity (r = 0.64, P

Background: Self-reported data are commonly used to estimate the prevalence of health conditions and the use of preventive health services in the population, but the validity of such data is often questioned. Methods: The Behavioral Risk Factor Survey (BRFS) was administered by telephone to a stratified, random sample of health maintenance organization (HMO) subscribers in Colorado in 1993, and self-reports were compared with HMO medical records for 599 adults aged >21. Sensitivity and specificity were calculated for three chronic conditions and use of six preventive services. Results: Sensitivity was highest for hypertension (83%), moderate for diabetes (73%), and lowest for hypercholesterolemia (59%); specificity was >80% for all three conditions. Sensitivity ranged from 86% to 99% for influenza immunization, clinical breast examination, blood cholesterol screening, mammography, Pap test, and blood pressure screening; specificity was Conclusions: Self-reports are reasonably accurate for certain chronic conditions and for routine screening exams and can provide a useful estimate for broad measures of population prevalence.

OBJECTIVES. The purpose of this study was to determine national trends in adolescent cigarette smoking prevalence. METHODS. We conducted trend analyses based on 1974 through 1991 current smoking prevalence data among persons aged 12 through 19 years from the National Household Surveys on Drug Abuse, High School Seniors Surveys, and National Health Interview Surveys. RESULTS. Overall smoking prevalence declined much more rapidly from 1974 through 1980 (1.9 percentage points annually among younger adolescents; the range among surveys of older adolescents was 0.2 to 2.0 percentage points annually) than from 1985 through 1991 (0 to 0.5 percentage points annually among all adolescents). Since 1980, smoking has generally declined at a slightly faster rate among older female adolescents than among male adolescents. Smoking among Black adolescents of all ages declined in nearly every survey population during each study period (range among surveys: 1974-1985 = 1.0 to 2.9 percentage points; 1985-1991 = 0.7 to 1.5 percentage points annually); for White adolescents, only minimal declines in smoking have occurred since 1985. CONCLUSIONS. Since 1974, major changes in adolescent smoking patterns have occurred, especially among Blacks. The overall slowing rate of decline in smoking prevalence since 1985 may indicate success of increased tobacco advertising and promotional activities targeted at adolescents or inadequate antitobacco education efforts.

Elevated blood pressure is a highly prevalent condition that is etiologically related to coronary heart disease and stroke, two of the leading causes of morbidity and mortality throughout the world. Excess salt (sodium chloride) intake is a major determinant of elevated blood pressure. In this article, we discuss the scientific rationale for population-wide salt reduction, the types and strength of available evidence, policy-making on dietary salt intake in the United States and other countries, and the role and impact of key stakeholders. We highlight a number of lessons learned, many of which are germane to policy development in other domains.

Clinical urine specimens are usually considered to be sterile when they do not yield uropathogens using standard clinical cultivation procedures. Our aim was to test if the adult female bladder might contain bacteria that are not identified by these routine procedures. An additional aim was to identify and recommend the appropriate urine collection method for the study of bacterial communities in the female bladder. Consenting participants who were free of known urinary tract infection provided urine samples by voided, transurethral, and/or suprapubic collection methods. The presence of bacteria in these samples was assessed by bacterial culture, light microscopy, and 16S rRNA gene sequencing. Bacteria that are not or cannot be routinely cultivated (hereinafter called uncultivated bacteria) were common in voided urine, urine collected by transurethral catheter (TUC), and urine collected by suprapubic aspirate (SPA), regardless of whether the subjects had urinary symptoms. Voided urine samples contained mixtures of urinary and genital tract bacteria. Communities identified in parallel urine samples collected by TUC and SPA were similar. Uncultivated bacteria are clearly present in the bladders of some women. It remains unclear if these bacteria are viable and/or if their presence is relevant to idiopathic urinary tract conditions.

Lactobacillus-dominated vaginal microbiotas are associated with reproductive health and STI resistance in women, whereas altered microbiotas are associated with bacterial vaginosis (BV), STI risk and poor reproductive outcomes. Putative vaginal taxa have been observed in male first-catch urine, urethral swab and coronal sulcus (CS) specimens but the significance of these observations is unclear. We used 16 S rRNA sequencing to characterize the microbiota of the CS and urine collected from 18 adolescent men over three consecutive months. CS microbiotas of most participants were more stable than their urine microbiotas and the composition of CS microbiotas were strongly influenced by circumcision. BV-associated taxa, including Atopobium, Megasphaera, Mobiluncus, Prevotella and Gemella, were detected in CS specimens from sexually experienced and inexperienced participants. In contrast, urine primarily contained taxa that were not abundant in CS specimens. Lactobacilllus and Streptococcus were major urine taxa but their abundance was inversely correlated. In contrast, Sneathia, Mycoplasma and Ureaplasma were only found in urine from sexually active participants. Thus, the CS and urine support stable and distinct bacterial communities. Finally, our results suggest that the penis and the urethra can be colonized by a variety of BV-associated taxa and that some of these colonizations result from partnered sexual activity.

To estimate the extent of childhood sexual abuse, questions were added to the 1993 Oregon Youth Risk Behavior Survey of students in grades 9-12. Twenty-five high schools throughout Oregon participated in the survey. Among the 2,332 students who answered sexual abuse questions, 20.9% had ever been sexually abused; females (33.1%) were much more likely to have ever been abused than were males (8.1%). Females (10.3%) also were more likely than males (3.4%) to have been sexually abused within the past year. High school students sexually abused in the past year engaged in many high risk health behaviors such as weapon carrying, substance abuse, seriously considering suicide in the past year, and sexual activity when compared with students who had never been sexually abused. Further educational efforts with teachers, health care providers, parents, and child care providers, as well as development and use of school curricula, are needed to reduce sexual abuse. Language: en

BACKGROUND. Although the disease consequences of cigarette smoking are well documented, smoking may also be associated with increased risk of injury. Our purpose is to provide an overview of this potential association. METHODS. We conducted a literature review. RESULTS. Cigarettes are the leading cause of death from fire and the second leading cause of fire-related injury. Studies estimate that compared with nonsmokers, smokers appear 1.5 times more likely to have a motor vehicle crash, 1.4-2.5 times more likely to be injured at work, and 2.0 times more likely to suffer other unintentional injuries. A variety of reasons may explain an association between cigarette smoking and injuries; these include (a) direct toxicity; (b) distractibility; (c) smoking-associated medical conditions; and (d) confounding factors, including personality or behavioral characteristics. CONCLUSIONS. Smoking may be an independent risk factor for thermal, motor vehicle, occupational, and other unintentional injuries. Nonsmokers may be at increased risk of injury from the presence of smokers in their environments, e.g., from fires. Societal benefits from decreased smoking prevalence are likely to include reduction of both fatal and nonfatal injuries. Language: en

The Summer Curriculum in Cancer Prevention has been sponsored by the National Cancer Institute’s Cancer Prevention Fellowship Program for over two decades. This curriculum includes a four-week course entitled “Principles and Practice of Cancer Prevention and Control.” The ultimate goal of this course is to present the most current cancer prevention research to a diverse workforce of researchers and practitioners eager to address the current challenges in this field. The course covers the current status of cancer prevention research and practice, ranging from epidemiology and clinical practice, and from basic to behavioral science research. It is comprised of lectures grouped into nine modules representing broad and specific topics relevant to cancer prevention. Course participants come from a broad cross-section of career stages, professions, and research interests, and are from across the United States and other countries. Over time and in response to feedback from participants, the course has developed to meet the needs and expectations of this diverse audience, and may serve as a model for those interested in cancer prevention education and training in other countries.

Project ID: 46317 – No publication moratorium.The microflora of the anterior male urethra during adolescence is poorly described and no data address the range of “typical” urethral organisms during adolescence as functions of pubertal development or onset of various types of partnered sexual activity. This means that current understanding of the urethral microbiome in adolescent men rests entirely on limited data inferred from adults, and from epidemiologic and behavioral explanations. Marked ethnic group variation in adolescent sexual behaviors and disparity in conditions such as sexually transmitted infections (STI) justifies exploration of potential ethnic group differences in the male urethral microbiome. Some research among adult males suggests important interactions between the male urethral microbiota and STI, but this research has not been extended to younger men.We will characterize the male urethral microbiome in longitudinal urine samples collected from a multi-ethnic cohort of adolescent males. The two specific aims of the project include Specific Aim 1. Describe the developmental urethral microbiome of adolescent males: Aim 1a. as a function of pubertal development; Aim 1b. as a function of racial/ethnic group; Aim 1c. as a function of oral-genital, vaginal, and anal sexual exposures; and, Aim 1d. as a function of circumcision status. Specific Aim 2. Describe variation in the urethral microbiome of adolescent males in the context of STI and identify organisms associated with acute and persistent non-specific urethritis: Aim 2a. to identify components of the microbiome associated with specific STI; Aim 2b. to compare the urethral microbiome of adolescent males who do and do not develop STI; and Aim 2c. to characterize microbial profiles associated with development of acute and persistent idiopathic urethritis.Microbial flora will be assessed as function of pubertal development, sexual activity, and incident STI. Subjects will be Latino (N=24), African-American (N=24), and Euro-American (N=24) males, ages 14 – 17 at enrollment; total N=72). Participants will be recruited from an urban community within an area of high prevalence of STI. Socio-demographic data will be collected at enrollment. Daily behavioral and symptomologic data will be collected via cell phone, which will also be used for cohort maintenance and scheduling of sample collections. Urine and penile coronal sulcus samples will be collected at enrollment and up to 36 1-month follow-up samples and selected contingent trigger samples triggered by specific symptoms and behaviors for characterization of microbiome using high-throughput 16s rRNA phylogeny, Q-PCR and metagenomic sequencing. STI testing by nucleic acid amplification will be done as well to better understand interactions between the urethral microbiome and infection. Cell phones also will be used to initiate alternate day sample collection for 14 days (7 samples) following trigger events such as specific sexual acts, antibiotic use, condom failure, or genital symptoms.

Health concerns have been pronounced for cosmetologists and manicurists, who are exposed daily to cosmetic products containing known or suspected human carcinogens and endocrine disruptors. In this retrospective cohort study, the authors used probabilistic record linkage between California's statewide cosmetology licensee and cancer surveillance files to identify newly diagnosed invasive cancers among female workforce members during 1988–2005. Rate ratios and 95% confidence intervals for cancer among workforce members compared with the general female population in California were estimated via Poisson regression. For comparison, site-specific proportional incidence ratios were computed. The authors identified 9,044 cancer cases in a cohort of 325,228 licensees. Rate ratios for all sites combined suggested lower incidence among both cosmetologists (rate ratio = 0.84, 95% confidence interval (CI): 0.82, 0.86) and manicurists (rate ratio = 0.87, 95% CI: 0.84, 0.90). Proportional incidence ratios were modestly elevated for thyroid cancer among all licensees (proportional incidence ratio = 1.13, 95% CI: 1.04, 1.23) and for lung cancer among manicurists (proportional incidence ratio = 1.21, 95% CI: 1.07, 1.36). Although there did not appear to be a cancer excess, these findings may be artifactually influenced by limitations in demographic information available from the licensee files. Additionally, the relatively young ages of cohort members and demographic shifts in the industry composition in recent years suggest a need for further follow-up.

Chlamydia trachomatis is the etiological agent of trachoma, the leading cause of preventable blindness. Trachoma presents distinct clinical syndromes ranging from mild and self-limiting to severe inflammatory disease. The underlying host and pathogen factors responsible for these diverse clinical outcomes are unclear. To assess the role played by pathogen variation in disease outcome, we analyzed the genomes of 4 trachoma strains representative of the 3 major trachoma serotypes, using microarray-based comparative genome sequencing. Outside of ompA, trachoma strains differed primarily in a very small subset of genes ([Formula: see text]). These subtle genetic variations were manifested in profound differences in virulence as measured by in vitro growth rate, burst size, plaque morphology, and interferon-gamma sensitivity but most importantly in virulence as shown by ocular infection of nonhuman primates. Our findings are the first to identify genes that correlate with differences in pathogenicity among trachoma strains.