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Berlin Brandenburg

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  • 1
    Language: English
    In: Proceedings of the National Academy of Sciences of the United States of America, 26 April 2011, Vol.108(17), pp.7189-93
    Description: Chlamydia trachomatis is an obligate intracellular bacterial pathogen that infects hundreds of millions of individuals globally, causing blinding trachoma and sexually transmitted disease. More effective chlamydial control measures are needed, but progress toward this end has been severely hampered by the lack of a tenable chlamydial genetic system. Here, we describe a reverse-genetic approach to create isogenic C. trachomatis mutants. C. trachomatis was subjected to low-level ethyl methanesulfonate mutagenesis to generate chlamydiae that contained less then one mutation per genome. Mutagenized organisms were expanded in small subpopulations that were screened for mutations by digesting denatured and reannealed PCR amplicons of the target gene with the mismatch specific endonuclease CEL I. Subpopulations with mutations were then sequenced for the target region and plaque-cloned if the desired mutation was detected. We demonstrate the utility of this approach by isolating a tryptophan synthase gene (trpB) null mutant that was otherwise isogenic to its parental clone as shown by de novo genome sequencing. The mutant was incapable of avoiding the anti-microbial effect of IFN-γ-induced tryptophan starvation. The ability to genetically manipulate chlamydiae is a major advancement that will enhance our understanding of chlamydial pathogenesis and accelerate the development of new anti-chlamydial therapeutic control measures. Additionally, this strategy could be applied to other medically important bacterial pathogens with no or difficult genetic systems.
    Keywords: Mutagenesis ; Mutation ; Chlamydia Trachomatis -- Genetics ; Tryptophan Synthase -- Genetics
    ISSN: 00278424
    E-ISSN: 1091-6490
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  • 2
    Language: English
    In: Cancer Research, 04/15/2011, Vol.71(8 Supplement), pp.2961-2961
    ISSN: 0008-5472
    E-ISSN: 1538-7445
    Source: CrossRef
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  • 3
    Language: English
    In: American Journal of Preventive Medicine, 2011, Vol.40(4), pp.419-426
    Description: Very little is known about the types of alcoholic beverages preferred by youth in the U.S. and the relationship between beverage preference and demographic and behavioral characteristics of these youth. To determine the type of alcoholic beverages consumed by adolescent drinkers and how it varies by drinking patterns. In 2010, an analysis was performed using 2007 data from the Youth Risk Behavior Survey (YRBS) conducted among public school students in eight states that included a question on the type of alcohol usually consumed. Analysis was restricted to the 7723 youth who reported consuming at least one drink of alcohol in the past 30 days. Beverage type preferences were analyzed by demographic factors, drinking patterns, and other health-risk behaviors. Logistic regression analyses were conducted to examine the correlates of type-specific alcohol consumption. Liquor was the strongly preferred alcoholic beverage of choice (43.8%), followed by beer (19.2%) and malt beverages (17.4%), with a very low preference for wine (3.7%) or wine coolers (3.4%). A higher preference for liquor or beer was observed among older youth, among those with a riskier pattern of alcohol consumption (e.g., greater frequency of consumption, binge drinking, or drinking and driving), and among youth who engaged in other risk behaviors. Riskier patterns of drinking and other health-risk behaviors are associated with an increased preference for hard liquor and beer. Improved surveillance of alcoholic beverage preferences among youth will enable a better understanding of the factors related to youth drinking, allowing the development of more effective interventions.
    Keywords: Medicine ; Public Health
    ISSN: 0749-3797
    E-ISSN: 1873-2607
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  • 4
    In: Addiction, September 2010, Vol.105(9), pp.1589-1596
    Description: Assess long‐term trends of the correlation between alcohol sales data and survey data. 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. United States. State alcohol tax authorities, alcohol vendors, alcohol industry (sales data) and randomly selected adults aged ≥ 18 years 1993–2006 (survey data). 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. 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. 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.
    Keywords: Alcohol Abuse ; Alcohol Drinking ; Cross‐Sectional Studies ; Drinking Behaviors ; Health Surveys ; Statistics
    ISSN: 0965-2140
    E-ISSN: 1360-0443
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  • 5
    Language: English
    In: Infection and immunity, August 2015, Vol.83(8), pp.3164-75
    Description: The ability of certain species of Chlamydia to inhibit the biogenesis of phagolysosomes permits their survival and replication within macrophages. The survival of macrophage-adapted chlamydiae correlates with the multiplicity of infection (MOI), and optimal chlamydial growth occurs in macrophages infected at an MOI of ≤1. In this study, we examined the replicative capacity of Chlamydia muridarum in the RAW 264.7 murine macrophage cell line at different MOIs. C. muridarum productively infected these macrophages at low MOIs but yielded few viable elementary bodies (EBs) when macrophages were infected at a moderate (10) or high (100) MOI. While high MOIs caused cytotoxicity and irreversible host cell death, macrophages infected at a moderate MOI did not show signs of cytotoxicity until late in the infectious cycle. Inhibition of host protein synthesis rescued C. muridarum in macrophages infected at a moderate MOI, implying that chlamydial growth was blocked by activated defense mechanisms. Conditioned medium from these macrophages was antichlamydial and contained elevated levels of interleukin 1β (IL-1β), IL-6, IL-10, and beta interferon (IFN-β). Macrophage activation depended on Toll-like receptor 2 (TLR2) signaling, and cytokine production required live, transcriptionally active chlamydiae. A hydroxyl radical scavenger and inhibitors of inducible nitric oxide synthase (iNOS) and cathepsin B also reversed chlamydial killing. High levels of reactive oxygen species (ROS) led to an increase in cathepsin B activity, and pharmacological inhibition of ROS and cathepsin B reduced iNOS expression. Our data demonstrate that MOI-dependent TLR2 activation of macrophages results in iNOS induction via a novel ROS- and cathepsin-dependent mechanism to facilitate C. muridarum clearance.
    Keywords: Cathepsin B -- Immunology ; Chlamydia Infections -- Immunology ; Chlamydia Muridarum -- Physiology ; Macrophages -- Enzymology ; Nitric Oxide -- Immunology ; Reactive Oxygen Species -- Immunology
    ISSN: 00199567
    E-ISSN: 1098-5522
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  • 6
    Language: English
    In: PLoS ONE, 01 January 2017, Vol.12(1), p.e0169859
    Description: Studies examining career satisfaction of biomedical scientists are limited, especially in the context of prior postdoctoral training. Here we focused on career satisfaction defined as satisfaction with one's career trajectory and perceived salary competitiveness among a predominantly Ph.D.-trained population of scientists who completed cancer prevention-related postdoctoral training between 1987-2011. National Cancer Institute (NCI) Cancer Prevention Fellowship Program (CPFP) alumni (n = 114), and previous recipients of NCI-sponsored Ruth L. Kirschstein National Research Service Award (NRSA/F32) postdoctoral fellowships (n = 140) completed online surveys. Associations of career satisfaction and perception of salary competitiveness with demographic, training, and employment-related factors were examined using logistic regression. Overall, 61% reported high levels of satisfaction with their career trajectory to-date. Higher salary (odds ratio [OR] = 2.86, 95% confidence interval [95% CI]: 1.07-7.69) and having more leadership roles (OR = 2.26, 95% CI:1.04-4.90) were independently associated with higher career satisfaction. Persons with race/ethnicity other than white (OR = 0.40, 95% CI: 0.20-0.82) or age ≥ 50 (OR = 0.40, 95%CI: 0.17-0.94) had lower career satisfaction levels. There were no statistically significant differences in career satisfaction levels by gender, scientific discipline, or employment sector. 74% perceived their current salary as competitive, but persons with 5-9, or ≥10 years in their current position reported lower levels (OR = 0.31, 95% CI: 0.15-0.65; and OR = 0.37, 95% CI: 0.16-0.87, respectively), as did individuals in government positions (OR = 0.33, 95% CI: 0.11-0.98). These data add to the understanding of career satisfaction of those with advanced training in biomedical research by examining these measures in relation to prior postdoctoral research training and across multiple career sectors.
    Keywords: Sciences (General)
    E-ISSN: 1932-6203
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  • 7
    Language: English
    In: American Journal of Preventive Medicine, 2011, Vol.40(4), pp.468-471
    Description: Binge drinking accounts for more than half of the 79,000 deaths due to excessive drinking in the U.S. each year. In 2006, the Behavioral Risk Factor Surveillance System (BRFSS) lowered the threshold for defining binge drinking among women from ≥5 drinks to ≥4 drinks per occasion, in accordance with national recommendations. To assess changes in binge-drinking prevalence among women. The relative and absolute change in binge drinking among U.S. adult women was assessed using pooled BRFSS data from the 2 years before (2004–2005) and after (2006–2007) the implementation of the new gender-specific definition. Analyses were conducted in 2008–2009. Binge-drinking prevalence among women increased 2.6 percentage points (from 7.3% in 2004–2005 to 9.9% in 2006–2007), a 35.6% relative increase. The percentage of women who reported consuming exactly 4 drinks in 2006 (3.6%) was similar to the increase in the prevalence of binge drinking among women that was observed from 2005 to 2006 (absolute change=2.9 percentage points). The new gender-specific definition of binge drinking significantly increased the identification of women drinking at dangerous levels. The change in prevalence among women was primarily due to the change in the definition and not to actual changes in drinking behavior. The new gender-specific definition of binge drinking can increase the usefulness of this measure for public health surveillance and support the planning and implementation of effective prevention strategies (e.g., increasing alcohol excise taxes).
    Keywords: Medicine ; Public Health
    ISSN: 0749-3797
    E-ISSN: 1873-2607
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  • 8
    Language: English
    In: PLoS One, San Francisco: Public Library of Science
    Description: Article discussing the microbial communities in male first catch urine and how these are highly similar to those paired in urethral swab specimens.
    Keywords: Microbials ; Bacteria ; Urine
    ISSN: 19326203
    E-ISSN: 19326203
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  • 9
    Language: English
    In: The Journal of biological chemistry, 22 July 2016, Vol.291(30), pp.15614-27
    Description: Cryptococcus neoformans (Cn) is a common facultative intracellular pathogen that can cause life-threatening fungal meningitis in immunocompromised individuals. Shortly after infection, Cn is detectable as both extra- and intracellular yeast particles, with Cn being capable of establishing long-lasting latent infections within host macrophages. Although recent studies have shown that shed capsular polysaccharides and intact extracellular Cn can compromise macrophage function through modulation of NF-κB signaling, it is currently unclear whether intracellular Cn also affects NF-κB signaling. Utilizing live cell imaging and computational modeling, we find that extra- and intracellular Cn support distinct modes of NF-κB signaling in cultured murine macrophages. Specifically, in RAW 264.7 murine macrophages treated with extracellular glucuronoxylomannan (GXM), the major Cn capsular polysaccharide, LPS-induced nuclear translocation of p65 is inhibited, whereas in cells with intracellular Cn, LPS-induced nuclear translocation of p65 is both amplified and sustained. Mathematical simulations and quantification of nascent protein expression indicate that this is a possible consequence of Cn-induced "translational interference," impeding IκBα resynthesis. We also show that long term Cn infection induces stable nuclear localization of p65 and IκBα proteins in the absence of additional pro-inflammatory stimuli. In this case, nuclear localization of p65 is not accompanied by TNFα or inducible NOS (iNOS) expression. These results demonstrate that capsular polysaccharides and intact intracellular yeast manipulate NF-κB via multiple distinct mechanisms and provide new insights into how Cn might modulate cellular signaling at different stages of an infection.
    Keywords: Cryptococcus Neoformans ; Nf-Kappa B (Nf-Kb) ; Computational Biology ; Host-Pathogen Interaction ; Inflammation ; Intracellular Pathogen ; Macrophage ; Models, Biological ; Cell Nucleus -- Metabolism ; Cryptococcosis -- Metabolism ; Cryptococcus Neoformans -- Metabolism ; Macrophages -- Metabolism ; Transcription Factor Rela -- Metabolism
    E-ISSN: 1083-351X
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  • 10
    Language: English
    In: American journal of public health, April 2013, Vol.103(4), pp.641-8
    Description: Our goal was to provide current estimates of alcohol-attributable cancer mortality and years of potential life lost (YPLL) in the United States. 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. 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. 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.
    Keywords: Alcohol Drinking -- Mortality ; Neoplasms -- Mortality
    ISSN: 00900036
    E-ISSN: 1541-0048
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