bluuebunny/crossref_metadata_2025_split huggingface dataset:
Salmonella Biofilm Development Depends on the Phosphorylation Status of RcsB
Authors: Latasa Cristina , García Begoña , Echeverz Maite , Toledo-Arana Alejandro , Valle Jaione , Campoy Susana , García-del Portillo Francisco , Solano Cristina , Lasa Iñigo 
Published: May 2012

ABSTRACT

The Rcs phosphorelay pathway is a complex signaling pathway involved in the regulation of many cell surface structures in enteric bacteria. In response to environmental stimuli, the sensor histidine kinase (RcsC) autophosphorylates and then transfers the phosphate through intermediary steps to the response regulator (RcsB), which, once phosphorylated, regulates gene expression. Here, we show that
Salmonella
biofilm development depends on the phosphorylation status of RcsB. Thus, unphosphorylated RcsB, hitherto assumed to be inactive, is essential to activate the expression of the biofilm matrix compounds. The prevention of RcsB phosphorylation either by the disruption of the phosphorelay at the RcsC or RcsD level or by the production of a nonphosphorylatable RcsB allele induces biofilm development. On the contrary, the phosphorylation of RcsB by the constitutive activation of the Rcs pathway inhibits biofilm development, an effect that can be counteracted by the introduction of a nonphosphorylatable RcsB allele. The inhibition of biofilm development by phosphorylated RcsB is due to the repression of CsgD expression, through a mechanism dependent on the accumulation of the small noncoding RNA RprA. Our results indicate that unphosphorylated RcsB plays an active role for integrating environmental signals and, more broadly, that RcsB phosphorylation acts as a key switch between planktonic and sessile life-styles in
Salmonella enterica
serovar Typhimurium.
Electron Bifurcation Involved in the Energy Metabolism of the Acetogenic Bacterium Moorella thermoacetica Growing on Glucose or H
2
plus CO
2
Authors: Huang Haiyan , Wang Shuning , Moll Johanna , Thauer Rudolf K. 
Published: May 2012

ABSTRACT

Moorella thermoacetica
ferments glucose to three acetic acids. In the oxidative part of the fermentation, the hexose is converted to 2 acetic acids and 2 CO
2
molecules with the formation of 2 NADH and 2 reduced ferredoxin (Fd
red
2−
) molecules. In the reductive part, 2 CO
2
molecules are reduced to acetic acid, consuming the 8 reducing equivalents generated in the oxidative part. An open question is how the two parts are electronically connected, since two of the four oxidoreductases involved in acetogenesis from CO
2
are NADP specific rather than NAD specific. We report here that the 2 NADPH molecules required for CO
2
reduction to acetic acid are generated by the reduction of 2 NADP
+
molecules with 1 NADH and 1 Fd
red
2−
catalyzed by the electron-bifurcating NADH-dependent reduced ferredoxin:NADP
+
oxidoreductase (NfnAB). The cytoplasmic iron-sulfur flavoprotein was heterologously produced in
Escherichia coli
, purified, and characterized. The purified enzyme was composed of 30-kDa (NfnA) and 50-kDa (NfnB) subunits in a 1-to-1 stoichiometry. NfnA harbors a [2Fe2S] cluster and flavin adenine dinucleotide (FAD), and NfnB harbors two [4Fe4S] clusters and FAD.
M. thermoacetica
contains a second electron-bifurcating enzyme. Cell extracts catalyzed the coupled reduction of NAD
+
and Fd with 2 H
2
molecules. The specific activity of this cytoplasmic enzyme was 3-fold higher in H
2
-CO
2
-grown cells than in glucose-grown cells. The function of this electron-bifurcating hydrogenase is not yet clear, since H
2
-CO
2
-grown cells additionally contain high specific activities of an NADP
+
-dependent hydrogenase that catalyzes the reduction of NADP
+
with H
2
. This activity is hardly detectable in glucose-grown cells.
Contributions of Individual σ
B
-Dependent General Stress Genes to Oxidative Stress Resistance of Bacillus subtilis
Authors: Reder Alexander , Höper Dirk , Gerth Ulf , Hecker Michael 
Published: May 2012

ABSTRACT

The general stress regulon of
Bacillus subtilis
comprises approximately 200 genes and is under the control of the alternative sigma factor σ
B
. The activation of σ
B
occurs in response to multiple physical stress stimuli as well as energy starvation conditions. The expression of the general stress proteins provides growing and stationary nonsporulating vegetative cells with nonspecific and broad stress resistance. A previous comprehensive phenotype screening analysis of 94 general stress gene mutants in response to severe growth-inhibiting stress stimuli, including ethanol, NaCl, heat, and cold, indicated that secondary oxidative stress may be a common component of severe physical stress. Here we tested the individual contributions of the same set of 94 mutants to the development of resistance against exposure to the superoxide-generating agent paraquat and hydrogen peroxide (H
2
O
2
). In fact, 62 mutants displayed significantly decreased survival rates in response to paraquat and/or H
2
O
2
stress compared to the wild type at a confidence level of an α value of ≤0.01. Thus, we were able to assign 47 general stress genes to survival against superoxide, 6 genes to protection from H
2
O
2
stress, and 9 genes to the survival against both. Furthermore, we show that a considerable overlap exists between the phenotype clusters previously assumed to be involved in oxidative stress management and the actual group of oxidative-stress-sensitive mutants. Our data provide information that many general stress proteins with still unknown functions are implicated in oxidative stress resistance and further support the notion that different severe physical stress stimuli elicit a common secondary oxidative stress.
Flavobacterium johnsoniae RemA Is a Mobile Cell Surface Lectin Involved in Gliding
Authors: Shrivastava Abhishek , Rhodes Ryan G. , Pochiraju Soumya , Nakane Daisuke , McBride Mark J. 
Published: May 2012

ABSTRACT

Cells of
Flavobacterium johnsoniae
move rapidly over surfaces by a process known as gliding motility. Gld proteins are thought to comprise the motor that propels the cell surface adhesin SprB. Cells with mutations in
sprB
are partially defective in motility and are also resistant to some bacteriophages. Transposon mutagenesis of a strain carrying a deletion spanning
sprB
identified eight mutants that were resistant to additional phages and exhibited reduced motility. Four of the mutants had transposon insertions in
remA
, which encodes a cell surface protein that has a lectin domain and appears to interact with polysaccharides. Three other genes identified in this screen (
remC
,
wza
, and
wzc
) encode proteins predicted to be involved in polysaccharide synthesis and secretion. Myc-tagged versions of RemA localized to the cell surface and were propelled rapidly along the cell at speeds of 1 to 2 μm/s. Deletion of
gldN
and
gldO
, which encode components of a bacteroidete protein secretion system, blocked the transport of RemA to the cell surface. Overexpression of RemA resulted in the formation of cell aggregates that were dispersed by the addition of galactose or rhamnose. Cells lacking RemC, Wza, and Wzc failed to aggregate. Cells of a
remC
mutant and cells of a
remA
mutant, neither of which formed aggregates in isolation, aggregated when they were mixed together, suggesting that polysaccharides secreted by one cell may interact with RemA on another cell. Fluorescently labeled lectin
Ricinus communis
agglutinin I detected polysaccharides secreted by
F. johnsoniae
. The polysaccharides bound to cells expressing RemA and were rapidly propelled on the cell surface. RemA appears to be a mobile cell surface adhesin, and secreted polysaccharides may interact with the lectin domain of RemA and enhance motility.
Autocatalytic Maturation of the Tat-Dependent Halophilic Subtilase Nep Produced by the Archaeon Natrialba magadii
Authors: Ruiz Diego M. , Paggi Roberto A. , Giménez Maria I. , De Castro Rosana E. 
Published: May 2012

ABSTRACT

Halolysins are subtilisin-like extracellular proteases produced by haloarchaea that possess unique protein domains and are salt dependent for structural integrity and functionality. In contrast to bacterial subtilases, the maturation mechanism of halolysins has not been addressed. The halolysin Nep is secreted by the alkaliphilic haloarchaeon
Natrialba magadii
, and the recombinant active enzyme has been synthesized in
Haloferax volcanii
. Nep contains an N-terminal signal peptide with the typical Tat consensus motif (G
RR
SVL), an N-terminal propeptide, the protease domain, and a C-terminal domain. In this study, we used Nep as a model protease to examine the secretion and maturation of halolysins by using genetic and biochemical approaches. Mutant variants of Nep were constructed by site-directed mutagenesis and expressed in
H. volcanii
, which were then analyzed by protease activity and Western blotting. The Tat dependence of Nep secretion was demonstrated in Nep RR/KK variants containing double lysine (KK) in place of the twin arginines (RR), in which Nep remained cell associated and the extracellular activity was undetectable. High-molecular-mass Nep polypeptides without protease activity were detected as cell associated and extracellularly in the Nep S/A variant, in which the catalytic serine 352 had been changed by alanine, indicating that Nep protease activity was needed for precursor processing and activation. Nep NSN 1-2 containing a modification in two potential cleavage sites for signal peptidase I (ASA) was not efficiently processed and activated. This study examined for the first time the secretion and maturation of a Tat-dependent halophilic subtilase.
Strategies to increase influenza vaccination rates: outcomes of a nationwide cross-sectional survey of UK general practice
Authors: Dexter Laura J , Teare M Dawn , Dexter Matthew , Siriwardena A Niroshan , Read Robert C 
Published: May 2012

ObjectiveTo identify practice strategies associated with higher flu vaccination rates in primary care.DesignLogistic regression analysis of data from a cross-sectional online questionnaire.Setting795 general practices across England.Participants569 practice managers, 335 nursing staff and 107 general practitioners.Primary outcome measuresFlu vaccination rates achieved by each practice in different groups of at-risk patients.Results7 independent factors associated with higher vaccine uptake were identified. Having a lead staff member for planning the flu campaign and producing a written report of practice performance predicted an 8% higher vaccination rate for at-risk patients aged <65 years (OR 1.37, 95% CI 1.10 to 1.71). These strategies, plus sending a personal invitation to all eligible patients and only stopping vaccination when Quality and Outcomes Framework targets are reached, predicted a 7% higher vaccination rate (OR 1.45, 95% CI 1.10 to 1.92) in patients aged ≥65 years. Using a lead member of staff for identifying eligible patients, with either a modified manufacturer's or in-house search programme for interrogating the practice IT system, independently predicted a 4% higher vaccination rate in patients aged ≥65 years (OR 1.22, 95% CI 1.06 to 1.41/OR 1.20, 95% CI 1.03 to 1.40). The provision of flu vaccine by midwives was associated with a 4% higher vaccination rate in pregnant women (OR 1.19, 95% CI 1.02 to 1.40).ConclusionsClear leadership, effective communication about performance and methods used to identify and contact eligible patients were independently associated with significantly higher rates of flu vaccination. Financial targets appear to incentivise practices to work harder to maximise seasonal influenza vaccine uptake. The strategies identified here could help primary care providers to substantially increase their seasonal flu vaccination rates towards or even above the Chief Medical Officer's targets.
Public health concerns for anti-obesity medicines imported for personal use through the internet: a cross-sectional study
Authors: Khan Mohiuddin Hussain , Tanimoto Tsuyoshi , Nakanishi Yoko , Yoshida Naoko , Tsuboi Hirohito , Kimura Kazuko 
Published: May 2012

ObjectiveTo explore the circulation of anti-obesity medicines via the internet and their quality.DesignCross-sectional study.SettingInternet pharmacies and pharmaceutical suppliers accessible from Japan.ParticipantsAnti-obesity medicines were purchased using relevant keywords on Japanese Google search engine. Blogs and advertisement-only sites were excluded.Primary and secondary outcome measuresThe authenticity of the samples was investigated in collaboration with the manufacturers of the samples and medicine regulatory authorities. Quality of the samples was assessed by pharmacopoeial analyses using high-performance liquid chromatography.Results82 samples were purchased from 36 internet sites. Approximately half of the sites did not mention a physical address, and 45% of the samples did not contain a package insert. A variety of custom declarations were made for the shipments of the samples: personal health items, supplement, medicines, general merchandise, tea and others. Among 82 samples, 52 samples were analysed to check their pharmacopoeial quality. Authenticity responses were received from only five of 20 manufacturing companies. According to the pharmacopoeial analyses and authenticity investigation, three of the samples were identified as counterfeits and did not contain any active ingredients. Two of these samples were confirmed as counterfeits by the manufacturer of the authentic products. The manufacturer of the other sample did not respond to our request for an authenticity check even after several communication attempts. These counterfeit cases have been reported at the rapid alert system of Western Pacific Region of the WHO.ConclusionsMany counterfeit and unapproved anti-obesity medicines may be easily bypassing regulatory checks during shipping and are widely circulated through the internet. Regulatory authorities should take measures to prevent these medicines from entering countries to safeguard their citizens.
Antidopaminergic drugs and acute pancreatitis: a population-based study
Authors: Bodén Robert , Bexelius Tomas S , Mattsson Fredrik , Lagergren Jesper , Lindblad Mats , Ljung Rickard 
Published: May 2012

ObjectivesTo evaluate the suggested association between antidopaminergic drugs and acute pancreatitis.DesignA large population-based nested case–control study.SettingSwedish nationwide study from 2006 to 2008.ParticipantsThe Patient Register was used to identify 6161 cases of acute pancreatitis. The 61 637 control subjects were randomly selected from the Register of the Total Population by frequency-based density sampling, matched for age, sex and calendar year.ExposureExposure data were extracted from the Prescribed Drug Register. Antidopaminergic drugs were grouped into antiemetic/anxiolytic and other antipsychotics. Current use of antidopaminergic drugs was defined as filling a prescription 1–114 days before index date, while previous use was 115 days to 3.5 years before index date.Main outcome measuresCases were defined as being diagnosed as having acute pancreatitis. ORs and 95% CIs were calculated using unconditional logistic regression.ResultsThe unadjusted OR indicated an increased risk of acute pancreatitis among current users of antiemetic/anxiolytics (OR 1.9, 95% CI 1.4 to 2.6), but not in the multivariable model adjusting for alcohol-related comorbidity, chronic obstructive lung disease, ischaemic heart disease, obesity, diabetes, opioid use, gallstone disease, educational level, marital status and number of concomitant medications (OR 0.9, 95% CI 0.6 to 1.2). Similarly, among current users of other antipsychotics, the unadjusted OR was 1.4 (95% CI 1.1 to 1.6), while the adjusted OR was 0.8 (95% CI 0.6 to 0.9). Results regarding previous use of antidopaminergic drugs followed a similar risk pattern as for current use.ConclusionsThe lack of association between antidopaminergic drugs and acute pancreatitis after adjustment for confounding factors in this study suggests that the previously reported positive associations might be explained by confounding.
Impact of obesity, overweight and underweight on life expectancy and lifetime medical expenditures: the Ohsaki Cohort Study
Authors: Nagai Masato , Kuriyama Shinichi , Kakizaki Masako , Ohmori-Matsuda Kaori , Sone Toshimasa , Hozawa Atsushi , Kawado Miyuki , Hashimoto Shuji , Tsuji Ichiro 
Published: May 2012

ObjectivesPeople who are obese have higher demands for medical care than those of the normal weight people. However, in view of their shorter life expectancy, it is unclear whether obese people have higher lifetime medical expenditure. We examined the association between body mass index, life expectancy and lifetime medical expenditure.DesignProspective cohort study using individual data from the Ohsaki Cohort Study.SettingMiyagi Prefecture, northeastern Japan.ParticipantsThe 41 965 participants aged 40–79 years.Primary and secondary outcome measuresThe life expectancy and lifetime medical expenditure aged from 40 years.ResultsIn spite of their shorter life expectancy, obese participants might require higher medical expenditure than normal weight participants. In men aged 40 years, multiadjusted life expectancy for those who were obese participants was 41.4 years (95% CI 38.28 to 44.70), which was 1.7 years non-significantly shorter than that for normal weight participants (p=0.3184). Multiadjusted lifetime medical expenditure for obese participants was £112 858.9 (94 954.1–131 840.9), being 14.7% non-significantly higher than that for normal weight participants (p=0.1141). In women aged 40 years, multiadjusted life expectancy for those who were obese participants was 49.2 years (46.14–52.59), which was 3.1 years non-significantly shorter than for normal weight participants (p=0.0724), and multiadjusted lifetime medical expenditure was £137 765.9 (123 672.9–152 970.2), being 21.6% significantly higher (p=0.0005).ConclusionsAccording to the point estimate, lifetime medical expenditure might appear to be higher for obese participants, despite their short life expectancy. With weight control, more people would enjoy their longevity with lower demands for medical care.
IL-10 Directly Activates and Expands Tumor-Resident CD8+ T Cells withoutDe NovoInfiltration from Secondary Lymphoid Organs
Authors: Emmerich Jan , Mumm John B. , Chan Ivan H. , LaFace Drake , Truong Hoa , McClanahan Terrill , Gorman Daniel M. , Oft Martin 
Published: May 2012

AbstractThe presence of activated intratumoral T cells correlates clinically with better prognosis in patients with cancer. Although tumor vaccines can increase the number of tumor-specific CD8+ T cells in systemic circulation, they frequently fail to increase the number of active and tumor reactive T cells within the tumor. Here we show that treatment with the pleiotropic cytokine interleukin-10 (IL-10) induces specific activation of tumor-resident CD8+ T cells as well as their intratumoral expansion in several mouse tumor models. We found that inhibition of T-cell trafficking from lymphoid organs did not impair IL-10–induced tumor rejection or the activation of tumor-resident CD8+ T cells. Tumor-resident CD8+ T cells expressed elevated levels of the IL-10 receptor and were directly activated by IL-10, resulting in prominent phosphorylation of STAT3 and STAT1. Although CD4+ T cells, regulatory T cells, NK cells, and dendritic cells have been reported as prominent targets of IL-10 in the tumor microenvironment, we found that expression of the IL-10R was required only on CD8+ T cells to facilitate IL-10–induced tumor rejection as well as in situ expansion and proliferation of tumor-resident CD8 T cells. Together, our findings indicate that IL-10 activates CD8+ T-cell–mediated tumor control and suggest that IL-10 may represent a potential tumor immunotherapy in human patients with cancer. Cancer Res; 72(14); 3570–81. ©2012 AACR.
Assessing Individual Breast Cancer Risk within the U.K. National Health Service Breast Screening Program: A New Paradigm for Cancer Prevention
Authors: Evans D. Gareth R. , Warwick Jane , Astley Susan M. , Stavrinos Paula , Sahin Sarah , Ingham Sarah , McBurney Helen , Eckersley Barbara , Harvie Michelle , Wilson Mary , Beetles Ursula , Warren Ruth , Hufton Alan , Sergeant Jamie C. , Newman William G. , Buchan Iain , Cuzick Jack , Howell Anthony 
Published: May 2012

Abstract
The aim of this study is to determine breast cancer risk at mammographic screening episodes and integrate standard risk factors with mammographic density and genetic data to assess changing the screening interval based on risk and offer women at high risk preventive strategies. We report our experience of assessing breast cancer risk within the U.K. National Health Service Breast Screening Program using results from the first 10,000 women entered into the "Predicting Risk Of breast Cancer At Screening" study. Of the first 28,849 women attending for screening at fifteen sites in Manchester 10,000 (35%) consented to study entry and completed the questionnaire. The median 10-year Tyrer–Cuzick breast cancer risk was 2.65% (interquartile range, 2.10–3.45). A total of 107 women (1.07%) had 10-year risks 8% or higher (high breast cancer risk), with a further 8.20% having moderately increased risk (5%–8%). Mammographic density (percent dense area) was 60% or more in 8.3% of women. We collected saliva samples from 478 women for genetic analysis and will extend this to 18% of participants. At time of consent to the study, 95.0% of women indicated they wished to know their risk. Women with a 10-year risk of 8% or more or 5% to 8% and mammographic density of 60% or higher were invited to attend or be telephoned to receive risk counseling; 81.9% of those wishing to know their risk have received risk counseling and 85.7% of these were found to be eligible for a risk-reducing intervention. These results confirm the feasibility of determining breast cancer risk and acting on the information in the context of population-based mammographic screening. Cancer Prev Res; 5(7); 943–51. ©2012 AACR.
Proprotein Convertase Subtilisin/Kexin Type 9 Interacts With Apolipoprotein B and Prevents Its Intracellular Degradation, Irrespective of the Low-Density Lipoprotein Receptor
Authors: Sun Hua , Samarghandi Amin , Zhang Ningyan , Yao Zemin , Xiong Momiao , Teng Ba-Bie 
Published: May 2012

Objective—
proprotein convertase subtilisin/kexin type 9 (PCSK9) negatively regulates the low-density lipoprotein (LDL) receptor (LDLR) in hepatocytes and therefore plays an important role in controlling circulating levels of LDL-cholesterol. To date, the relationship between PCSK9 and metabolism of apolipoprotein B (apoB), the structural protein of LDL, has been controversial and remains to be clarified.

Methods and Results—

We assessed the impact of PCSK9 overexpression (≈400-fold above baseline) on apoB synthesis and secretion in 3 mouse models: wild-type C57BL/6 mice and LDLR-null mice (
Ldlr
−/−
and
Ldlr
−/−
Apobec1
−/−
). Irrespective of LDLR expression, mice transduced with the
PCSK9
gene invariably exhibited increased levels of plasma cholesterol, triacylglycerol, and apoB. Consistent with these findings, the levels of very-low-density lipoprotein and LDL were also increased whereas high-density lipoprotein levels were unchanged. Importantly, we demonstrated that endogenous PCSK9 interacted with apoB in hepatocytes. The PCSK9/apoB interaction resulted in increased production of apoB, possibly through the inhibition of intracellular apoB degradation via the autophagosome/lysosome pathway.

Conclusion—
We propose a new role for PCSK9 that involves shuttling between apoB and LDLR. The present study thus provides new insights into the action of PCSK9 in regulating apoB metabolism. Furthermore, our results indicate that targeting PCSK9 expression represents a new paradigm in therapeutic intervention against hyperlipidemia.