bluuebunny/crossref_metadata_2025 huggingface dataset:
Identification of tyrosylprotein sulfotransferase in
Arabidopsis
Authors: Komori Ryota , Amano Yukari , Ogawa-Ohnishi Mari , Matsubayashi Yoshikatsu
Published: July 2009

Tyrosine sulfation is a posttranslational modification common in peptides and proteins synthesized by the secretory pathway in most eukaryotes. In plants, this modification is critical for the biological activities of a subset of peptide hormones such as PSK and PSY1. In animals, tyrosine sulfation is catalyzed by Golgi-localized type II transmembrane proteins called tyrosylprotein sulfotransferases (TPSTs). However, no orthologs of animal TPST genes have been found in plants, suggesting that plants have evolved plant-specific TPSTs structurally distinct from their animal counterparts. To investigate the mechanisms of tyrosine sulfation in plants, we purified TPST activity from microsomal fractions of
Arabidopsis
MM2d cells, and identified a 62-kDa protein that specifically interacts with the sulfation motif of PSY1 precursor peptide. This protein is a 500-aa type I transmembrane protein that shows no sequence similarity to animal TPSTs. A recombinant version of this protein expressed in yeast catalyzed tyrosine sulfation of both PSY1 and PSK precursor polypeptide in vitro, indicating that the newly identified protein is indeed an
Arabidopsis
(At)TPST. AtTPST is expressed throughout the plant body, and the highest levels of expression are in the root apical meristem. A loss-of-function mutant of
AtTPST
displayed a marked dwarf phenotype accompanied by stunted roots, pale green leaves, reduction in higher order veins, early senescence, and a reduced number of flowers and siliques. Our results indicate that plants and animals independently acquired tyrosine sulfation enzymes through convergent evolution.
TR4 nuclear receptor functions as a fatty acid sensor to modulate CD36 expression and foam cell formation
Authors: Xie Shaozhen , Lee Yi-Fen , Kim Eungseok , Chen Lu-Min , Ni Jing , Fang Lei-Ya , Liu Su , Lin Shin-Jen , Abe Jun-ichi , Berk Bradford , Ho Feng-Ming , Chang Chawnshang
Published: July 2009

Testicular orphan nuclear receptor 4 (TR4) is an orphan member of the nuclear receptor superfamily with diverse physiological functions. Using TR4 knockout (TR4
−/−
) mice to study its function in cardiovascular diseases, we found reduced cluster of differentiation (CD)36 expression with reduced foam cell formation in TR4
−/−
mice. Mechanistic dissection suggests that TR4 induces CD36 protein and mRNA expression via a transcriptional regulation. Interestingly, we found this TR4-mediated CD36 transactivation can be further enhanced by polyunsaturated fatty acids (PUFAs), such as omega-3 and -6 fatty acids, and their metabolites such as 15-hydroxyeico-satetraonic acid (15-HETE) and 13-hydroxy octa-deca dieonic acid (13-HODE) and thiazolidinedione (TZD)-rosiglitazone. Both electrophoretic mobility shift assays (EMSA) and chromatin immunoprecipitation (ChIP) assays demonstrate that TR4 binds to the TR4 response element located on the CD36 5′-promoter region for the induction of CD36 expression. Stably transfected TR4-siRNA or functional TR4 cDNA in the RAW264.7 macrophage cells resulted in either decreased or increased CD36 expression with decreased or increased foam cell formation. Restoring functional CD36 cDNA in the TR4 knockdown macrophage cells reversed the decreased foam cell formation. Together, these results reveal an important signaling pathway controlling CD36-mediated foam cell formation/cardiovascular diseases, and findings that TR4 transactivation can be activated via its ligands/activators, such as PUFA metabolites and TZD, may provide a platform to screen new drug(s) to battle the metabolism syndrome, diabetes, and cardiovascular diseases.
Inhibition of
aac(6′)-Ib
-mediated amikacin resistance by nuclease-resistant external guide sequences in bacteria
Authors: Soler Bistué Alfonso J. C. , Martín Fernando A. , Vozza Nicolás , Ha Hongphuc , Joaquín Jonathan C. , Zorreguieta Angeles , Tolmasky Marcelo E.
Published: July 2009

Inhibition of bacterial gene expression by RNase P-directed cleavage is a promising strategy for the development of antibiotics and pharmacological agents that prevent expression of antibiotic resistance. The rise in multiresistant bacteria harboring AAC(6′)-Ib has seriously limited the effectiveness of amikacin and other aminoglycosides. We have recently shown that recombinant plasmids coding for external guide sequences (EGS), short antisense oligoribonucleotides (ORN) that elicit RNase P-mediated cleavage of a target mRNA, induce inhibition of expression of
aac(6′)-Ib
and concomitantly induce a significant decrease in the levels of resistance to amikacin. However, since ORN are rapidly degraded by nucleases, development of a viable RNase P-based antisense technology requires the design of nuclease-resistant RNA analog EGSs. We have assayed a variety of ORN analogs of which selected LNA/DNA co-oligomers elicited RNase P-mediated cleavage of mRNA in vitro. Although we found an ideal configuration of LNA/DNA residues, there seems not to be a correlation between number of LNA substitutions and level of activity. Exogenous administration of as low as 50 nM of an LNA/DNA co-oligomer to the hyperpermeable
E. coli
AS19 harboring the
aac(6′)-Ib
inhibited growth in the presence of amikacin. Our experiments strongly suggest an RNase P-mediated mechanism in the observed antisense effect.
The molecular basis for impaired hypoxia-induced VEGF expression in diabetic tissues
Authors: Thangarajah Hariharan , Yao Dachun , Chang Edward I. , Shi Yubin , Jazayeri Leila , Vial Ivan N. , Galiano Robert D. , Du Xue-Liang , Grogan Raymon , Galvez Michael G. , Januszyk Michael , Brownlee Michael , Gurtner Geoffrey C.
Published: July 2009

Diabetes is associated with poor outcomes following acute vascular occlusive events. This results in part from a failure to form adequate compensatory microvasculature in response to ischemia. Since vascular endothelial growth factor (VEGF) is an essential mediator of neovascularization, we examined whether hypoxic up-regulation of VEGF was impaired in diabetes. Both fibroblasts isolated from type 2 diabetic patients, and normal fibroblasts exposed chronically to high glucose, were defective in their capacity to up-regulate VEGF in response to hypoxia. In vivo, diabetic animals demonstrated an impaired ability to increase VEGF production in response to soft tissue ischemia. This resulted from a high glucose-induced decrease in transactivation by the transcription factor hypoxia-inducible factor-1α (HIF-1α), which mediates hypoxia-stimulated VEGF expression. Decreased HIF-1α functional activity was specifically caused by impaired HIF-1α binding to the coactivator p300. We identify covalent modification of p300 by the dicarbonyl metabolite methylglyoxal as being responsible for this decreased association. Administration of deferoxamine abrogated methylglyoxal conjugation, normalizing both HIF-1α/p300 interaction and transactivation by HIF-1α. In diabetic mice, deferoxamine promoted neovascularization and enhanced wound healing. These findings define molecular defects that underlie impaired VEGF production in diabetic tissues and offer a promising direction for therapeutic intervention.
Negative membrane curvature as a cue for subcellular localization of a bacterial protein
Authors: Ramamurthi Kumaran S. , Losick Richard
Published: July 2009

Bacterial proteins often localize to distinct sites within the cell, but the primary cues that dictate localization are largely unknown. Recent evidence has shown that positive membrane curvature can serve as a cue for localization of a peripheral membrane protein. Here we report that localization of the peripheral membrane protein DivIVA is determined in whole or in part by recognition of negative membrane curvature and that regions of the protein near the N and C terminus are important for localization. DivIVA, which is a cell division protein in
Bacillus subtilis
, localizes principally as a ring at nascent septa and secondarily to the less negatively curved, inside surface of the hemispherical poles of the cell. When cytokinesis is prevented, DivIVA redistributes itself to, and becomes markedly enriched at, the poles. When the rod-shaped cells are converted into spheres (protoplasts) by treatment with lysozyme, DivIVA adopts a largely uniform distribution around the cell. Recognition of membrane curvature by peripheral membrane proteins could be a general strategy for protein localization in bacteria.
Patients' Beliefs and Perceptions of Their Participation to Increase Healthcare Worker Compliance with Hand Hygiene
Authors: Longtin Yves , Sax Hugo , Allegranzi Benedetta , Hugonnet Stéphane , Pittet Didier
Published: July 2009

Background.

Research suggests that patients could improve healthcare workers' compliance with hand hygiene recommendations by reminding them to cleanse their hands.

Objective.

To assess patients' perceptions of a patient-participation program to improve healthcare workers' compliance with hand hygiene.

Design.

Cross-sectional survey of patient knowledge and perceptions of healthcare-associated infections, hand hygiene, and patient participation, defined as the active involvement of patients in various aspects of their health care.

Setting.

Large Swiss teaching hospital.

Results.

Of 194 patients who participated, most responded that they would not feel comfortable asking a nurse (148 respondents [76%]) or a physician (150 [77%]) to perform hand hygiene, and 57 (29%) believed that this would help prevent healthcare-associated infections. In contrast, an explicit invitation from a healthcare worker to ask about hand hygiene doubled the intention to ask a nurse (from 34% to 83% of respondents; P < .001) and to ask a physician (from 30% to 78%; P < .001). In multivariate analysis, being nonreligious, having an expansive personality, being concerned about healthcare-associated infections, and believing that patient participation would prevent healthcare-associated infections were associated with the intention to ask a nurse or a physician to perform hand hygiene (P < .05). Being of Jewish, Eastern Orthodox, or Buddhist faith was associated also with increased intention to ask a nurse (P < .05), compared with being of Christian faith.

Conclusions.

This study identifies several sociodemographic characteristics associated with the intention to ask nurses and physicians about hand hygiene and underscores the importance of a direct invitation from healthcare workers to increase patient participation and foster patient empowerment. These findings could guide the development of future hand hygiene-promotion strategies.
Primary Care Guidelines for the Management of Persons Infected with Human Immunodeficiency Virus: 2009 Update by the HIV Medicine Association of the Infectious Diseases Society of America
Authors: Aberg Judith A. , Kaplan Jonathan E. , Libman Howard , Emmanuel Patricia , Anderson Jean R. , Stone Valerie E. , Oleske James M. , Currier Judith S. , Gallant Joel E.
Published: July 2009

Evidence-based guidelines for the management of persons infected with human immunodeficiency virus (HIV) were prepared by an expert panel of the HIV Medicine Association of the Infectious Diseases Society of America. These updated guidelines replace those published in 2004. The guidelines are intended for use by health care providers who care for HIV-infected patients or patients who may be at risk for acquiring HIV infection. Since 2004, new antiretroviral drugs and classes have become available, and the prognosis of persons with HIV infection continues to improve. However, with fewer complications and increased survival, HIV-infected persons are increasingly developing common health problems that also affect the general population. Some of these conditions may be related to HIV infection itself and its treatment. HIV-infected persons should be managed and monitored for all relevant age- and gender-specific health problems. New information based on publications from the period 2003–2008 has been incorporated into this document.
Progress in the development of deposition prevention and cleaning techniques of in-vessel optics in ITER
Authors: Mukhin E. , Vukolov K. , Semenov V. , Tolstyakov S. , Kochergin M. , Kurskiev G. , Podushnikova K. , Razdobarin A. , Gorodetsky A. , Zalavutdinov R. , Bukhovets V. , Zakharov A. , Bulovich S. , Veiko V. , Shakshno E.
Published: July 2009

The lifetime of front optical components unprotected from reactor grade plasmas may be very short due to intensive contamination with carbon and beryllium-based materials eroded by the plasma from beryllium walls and carbon tiles. Deposits result in a significant reduction and spectral alterations of optical transmission. In addition, even rather thin and transparent deposits can dramatically change the shape of reflectance spectra, especially for mirrors with rather low reflectivity, such as W or Mo. The distortion of data obtained with various optical diagnostics may affect the safe operation of ITER. Therefore, the development of optics-cleaning and deposition-mitigating techniques is a key factor in the construction and operation of optical diagnostics in ITER. The problem is of particular concern for optical elements positioned in the divertor region. The latest achievements in protection of in-vessel optics are presented using the example of deposition prevention/cleaning techniques for in-machine components of the Thomson scattering system in the divertor. Careful consideration of well-known and novel protection approaches shows that neither of them alone provides guaranteed survivability of the first in-vessel optics in the divertor. Only a set of complementary prevention/cleaning techniques, which include special materials for mirrors and inhibition additives for plasma, is able to manage the challenging task. The essential issue, which needs to be addressed in the immediate future, is an extensive development of techniques tested under experimental conditions (exposure time and contamination fluxes) similar to those expected in ITER.
Progress towards a predictive model for pedestal height in DIII-D
Authors: Groebner R.J. , Leonard A.W. , Snyder P.B. , Osborne T.H. , Maggi C.F. , Fenstermacher M.E. , Petty C.C. , Owen L.W.
Published: July 2009

Recent DIII-D pedestal studies provide improved characterization of pedestal scaling for comparison with models. A new pedestal model accurately predicts the maximum achieved pedestal width and height in type I ELMing discharges over a large range of DIII-D operational space, including ITER demonstration discharges. The model is a combination of the peeling–ballooning theory for the MHD stability limits on the pedestal with a simple pedestal width scaling in which the width is proportional to the square root of the pedestal poloidal beta. Width scalings based on the ion toroidal or poloidal gyroradius are much poorer descriptions of DIII-D data. A mass scaling experiment in H and D provides support for a poloidal beta scaling and is not consistent with an ion poloidal gyroradius scaling. Studies of pedestal evolution during the inter-ELM cycle provide evidence that both the pedestal width and height increase during pedestal buildup. Model studies with a 1D kinetic neutrals calculation show that the temporal increase in density width cannot be explained in terms of increased neutral penetration depth. These studies show a correlation of pedestal width with both the square root of the pedestal poloidal beta and the square root of the pedestal ion temperature during the pedestal buildup.
Particle Size Effects on the Quality of Flour Tortillas Enriched with Whole Grain Waxy Barley
Authors: Prasopsunwattana Naruemon , Omary María Botero , Arndt Elizabeth A. , Cooke Peter H. , Flores Rolando A. , Yokoyama Wallace , Toma Ayako , Chongcham Sutida , Lee Sylvia P.
Published: July 2009

Wheat tortillas were enriched with whole barley flour (WBF) of different particle sizes including 237 μm (regular [R]), 131 μm (intermediate [IM]), and 68 μm (microground [MG]). Topographical and fluorescent microstructure images of flours, doughs, and tortillas were examined. Flours and tortillas were analyzed for color, protein, ash, starch, moisture, and β‐glucan content. Farinograph testing was conducted on the flour blends. Water activity and texture analyses of tortillas were conducted. A 9‐point hedonic scale was used by 95 untrained panelists to evaluate tortilla appearance, color, flavor, texture, and overall acceptability. Two commercial products (CP) were included in some analyses. As WBF particle size decreased, color was lighter; protein, moisture content and mixing stability decreased; ash, starch content, water absorption and farinograph peak time increased; and β‐glucan content was constant. WBF tortillas were darker than the control (C), while IM and MG tortillas had lower peak forces than C. No flavor differences were reported among C, R, and MG tortillas but higher scores were given to both CP in all attributes tested. Tortillas made with the largest WBF particle size (R) were the most similar in protein content, texture and flavor when compared with C tortillas made with refined bread flour.
Impact of Genotype and Environment on the Quality of Amber Durum Wheat Alkaline Noodles
Authors: Hatcher D. W. , Dexter J. E. , Bellido G. G. , Clarke J. M. , Anderson M. J.
Published: July 2009

Canada Western Amber Durum wheat cultivars (4), Canada Western Red Spring (1), and Canada Western Hard White Spring (1) wheat were grown at three sites in 2007 to evaluate the effect of genotype (G) and environment (E) on the quality of yellow alkaline noodles (YAN). YAN were evaluated for color, appearance, and cooked texture. Brightness (L*) and yellowness (b*) of YAN made from durum cultivars were significantly higher than common wheat. Durum flour yellow pigment content was approximately fourfold greater than common wheat while noodle speckiness was approximately half of CWRS at 2 hr with environment accounting for >75% of the variance for each parameter. Resistance to compression (RTC) and recovery (REC) of cooked durum alkaline noodles were equivalent or superior to common wheat noodles even when lower grade durum wheat flour was used. In conclusion, cooked durum noodle texture parameters were all significantly influenced by genotype and environment, with environment accounting for 66–71% of their variance.
Size‐Exclusion HPLC of Protein Using a Narrow‐Bore Column for Evaluation of Breadmaking Quality of Hard Spring Wheat Flours
Authors: Ohm Jae‐Bom , Hareland Gary , Simsek Senay , Seabourn Bradford
Published: July 2009

The objective of this study was to investigate whether a narrow‐bore column (NBC) (300 × 4.5 mm, i.d.) improved analyses of unreduced proteins in flour by size‐exclusion HPLC (SE‐HPLC) and subsequent evaluation of breadmaking quality of hard spring wheat flours. Total protein extracts and SDS buffer extractable and unextractable proteins were analyzed by SE‐HPLC. NBC separated proteins in 10 min at a flow rate of 0.5 mL/min with similar resolution to a regular column (300 × 7.8 mm, i.d.) which took 30 min. SE‐HPLC absorbance area (AA) data obtained from an NBC showed comparable or superior repeatability and correlations with flour breadmaking characteristics when compared with those of a regular column. AA values of total protein that were calculated by adding AA values of SDS extractable and unextractable proteins showed greater repeatability and correlations with quality characteristics than those of actual total protein extracts. The improvements including employment of an NBC in SE‐HPLC provide enhancement of rapid quality evaluation and decreased consumption of hazardous organic solvents.
A Comparative Study Between the McGill #2 Laboratory Mill and Commercial Milling Systems
Authors: Graves A. M. , Siebenmorgen T. J. , Saleh M. I.
Published: July 2009

The degree of similarity between rice milled in a McGill #2 laboratory mill and commercial milling processes was evaluated using eight physical, physicochemical, and end‐use properties. There was no statistical difference between the two milling systems with respect to color parameters L* and a*, final viscosity, texture, and end‐use cooking properties (α = 0.05). Overall, the kernel dimensions of length, width, and thickness were less in the McGill #2 laboratory‐milled rice than the same rice milled commercially. The incidence of bran streaks and peak viscosity values were each higher when the rice sample was milled commercially in 27, and 28, respectively, of the 29 samples by means comparison. The decrease in kernel dimensions and incidence of bran streaks were attributed to the more aggressive nature of the single‐pass, batch milling system of the McGill #2 laboratory mill as compared with multipass, continuous milling systems that are used commercially. Finally, as surface lipid content (SLC) decreased, L* increased and a*, b*, and the incidence of bran streaks decreased for both milling systems.
Effects of Flour Extraction Rate, Added Water, and Salt on Color and Texture of Chinese White Noodles
Authors: Ye Yili , Zhang Yan , Yan Jun , Zhang Yong , He Zhonghu , Huang Sidi , Quail Kenneth J.
Published: July 2009

Both cultivar and noodle composition and preparation have important effects on noodle quality. In this study, the effects of flour extraction rate (50, 60, and 70%), added water (33, 35, and 37%), and salt concentration (0, 1, and 2%, w/w) on color and texture of Chinese white noodle (CWN) were investigated using flour samples from five leading Chinese wheat cultivars. The five samples showed large variations in protein content, ash content, flour color, farinograph, and extensigraph parameters, and starch pasting properties. Analyses of variance indicated that cultivar, flour extraction rate, level of water addition, salt concentration, and the interactions had significant effects on color of raw noodle sheets and color and textural properties of CWN. Cultivar and water addition were more important sources of variation than flour extraction rate and salt concentration. The brightness (L*) and redness (a*) values of raw noodle sheets were significantly reduced and increased, respectively, as flour extraction rate increased from 50 to 70%, and noodle scores were slightly higher at flour extraction rates of 50%. Water addition showed different effects on raw noodle sheet color at 2 and 24 hr, and a significant improvement was observed for noodle appearance, firmness, viscoelasticity, smoothness, and total score as water addition increased from 33 to 37%. L* of raw noodle sheets, and firmness and viscoelasticity of cooked noodles, were significantly improved, but noodle flavor significantly deteriorated as salt concentration increased from 0 to 2%; 1% salt produced the highest noodle score. Thus, the recommended composition for laboratory preparation of CWN is 60% flour extraction, 35% water addition, and 1% salt concentration.
Renal function predicts cardiovascular outcomes in southern Italian postmenopausal women
Authors: Perticone Francesco , Sciacqua Angela , Maio Raffaele , Perticone Maria , Laino Irma , Bruni Rosamaria , Cello Serena Di , Leone Giulia Galiano , Greco Laura , Andreozzi Francesco , Sesti Giorgio
Published: July 2009

Background

Postmenopausal women have an increased risk of adverse cardiovascular (CV) events. Similarly, chronic kidney disease (CKD) is a well established risk factor for CV disease and mortality.

Design

We evaluated the effect of renal function on the risk of death and CV events in 1500 southern Italian postmenopausal women.

Methods and results

Renal function was estimated (e) by glomerular filtration rate (e-GFR) by Modification of Diet in Renal Disease equation. We classified postmenopausal women in two groups of e-GFR (ml/min per 1.73 m2): ≥ 60 (group 1) and less than 60 (group 2). The primary endpoint was major adverse CV events (MACE). The secondary endpoints were total events (MACE + death from any cause), coronary events, and stroke. During the follow-up (mean = 72.6 months), there were 200 new CV morbid events. The rate of MACE (per 100 patient-years) was 1.88 and 2.98 in the two groups of e-GFR ( P < 0.0001). On univariate analysis, the incident risk of CV events was inversely related with the e-GFR values; similarly, in multiple Cox regression model, only the e-GFR maintained an independent association with MACE and secondary end-points.

Conclusion

For the first time, we demonstrated that the reduction of e-GFR was associated with the increased risk of death and CV events, independently of traditional CV risk factors, menopause duration, and presence of metabolic syndrome.
Vitality and recurrent event risk in acute myocardial infarction survivors
Authors: Williams Brent A. , Dorn Joan M. , Donahue Richard P. , Hovey Kathleen M. , Rafalson Lisa B. , Trevisan Maurizio
Published: July 2009

Background

Low vitality, characterized by fatigue and lack of energy, is common among survivors of acute myocardial infarction (AMI) and has been shown to be associated with increased risk of primary and secondary cardiac events. The goal of this study was to determine whether an association between vitality and recurrent cardiac events (nonfatal MI, cardiac death) among acute MI survivors persists after controlling for possible physiological and psychological confounders.

Design and methods

Incident AMI survivors ( n = 1328) from Erie and Niagara (New York) county hospitals were enrolled and followed up to 9 years. Vitality was measured by the Short Form-36 on a 0–100 scale approximately 4 months post-AMI. Cox proportional hazards models were developed to assess the vitality-recurrent event association controlling for traditional cardiovascular disease risk factors, index MI severity, and psychological correlates of vitality.

Results

Low-vitality individuals at baseline were more likely females, of higher BMI, smoking, diabetic, less physically active, and to have worse depression scores. Vitality was not strongly associated with MI severity markers. Lower vitality scores were associated with increased risk of recurrent cardiac events: adjusted hazard ratios (95% CI) for vitality scores 51–79, 21–50, and ≤ 20 (compared with ≥ 80) were 1.2 (0.8, 1.8), 1.4 (0.9, 2.2), and 2.9 (1.5, 5.4), respectively ( Ptrend = 0.005).

Conclusion

Low vitality was associated with increased risk of recurrent cardiac events among AMI survivors after controlling for physiological and psychological confounders. Mechanistic links with vitality should be sought as interventional targets.
Smoking as an independent risk factor for myocardial infarction or stroke in type 2 diabetes: a report from the Swedish National Diabetes Register
Authors: Nilsson Peter M. , Cederholm Jan , Eeg-Olofsson Katarina , Eliasson Björn , Zethelius Björn , Fagard Robert , rnsdóttir Soffia Gudbjö ,   for the Swedish National Diabetes Register
Published: July 2009

Background

Few earlier studies have analysed smoking as a risk factor for myocardial infarction (MI) or stroke in type 2 diabetic patients.

Design and methods

A longitudinal study involved 13 087 female and male patients with type 2 diabetes from the Swedish National Diabetes Register with no previous MI or stroke at baseline, aged 30–74 years, and with data available for all analysed variables, followed up for mean 5.7 years.

Results

Adjusted hazard ratios (HRs) for smoking and first-incident fatal/nonfatal MI, stroke and total mortality were 1.7 [95% confidence interval (CI): 1.4–2.0; P < 0.001], 1.3 (95% CI: 1.1–1.6; P = 0.006) and 1.8 (95% CI: 1.5–2.2; P < 0.001), respectively, by Cox regression analysis, adjusted for age, sex, diabetes duration, hypoglycaemic treatment, haemoglobin A1c, blood pressure, body mass index, microalbuminuria, antihypertensive and lipid-lowering drugs. Adjusted HR was higher for fatal MI, 2.1 (95% CI: 1.7–2.7; P < 0.001), than for nonfatal MI, 1.4 (95% CI: 1.2–1.7; P < 0.001). The highest HRs were observed in more frequently smoking (22%), middle-aged patients (age < 60 years) for fatal/nonfatal MI, 2.3 (95% CI: 1.8–3.1; P < 0.001) and for total mortality, 2.5 (95% CI: 1.6–3.8, P < 0.001), whereas lower HRs were observed in older and less smoking patients. With predicted cessation of smoking in patients aged below 60 years, 24% (95% CI: 15–33%) of cases of fatal/nonfatal MI and 24% (11–37%) of cases of total mortality may have been prevented.

Conclusion

The risk for MI and total mortality associated with smoking is high in type 2 diabetes, especially in more frequently smoking, middle-aged patients, and was higher for MI than for stroke, and also higher for fatal than for nonfatal events. Smoking cessation would strongly affect risk reduction.
On the reliability of earth slopes in three dimensions
Authors: Griffiths D. V. , Huang Jinsong , Fenton Gordon A.
Published: July 2009

The paper investigates the probability of failure of two-dimensional and three-dimensional slopes using the random finite-element method (RFEM). In this context, RFEM combines elastoplastic finite-element algorithms with random field theory in a Monte Carlo framework. Full account is taken of local averaging and variance reduction over each element, and an exponentially decaying (Markov) spatial correlation function is incorporated. It is found that two-dimensional probabilistic analysis, which implicitly assumes perfect spatial correlation in the out-of-plane direction, may underestimate the probability of failure of slopes.
Velvet worm development links myriapods with chelicerates
Authors: Mayer Georg , Whitington Paul M.
Published: July 2009

Despite the advent of modern molecular and computational methods, the phylogeny of the four major arthropod groups (Chelicerata, Myriapoda, Crustacea and Hexapoda, including the insects) remains enigmatic. One particular challenge is the position of myriapods as either the closest relatives to chelicerates (Paradoxopoda/Myriochelata hypothesis), or to crustaceans and hexapods (Mandibulata hypothesis). While neither hypothesis receives conclusive support from molecular analyses, most morphological studies favour the Mandibulata concept, with the mandible being the most prominent feature of this group. Although no morphological evidence was initially available to support the Paradoxopoda hypothesis, a putative synapomorphy of chelicerates and myriapods has recently been put forward based on studies of neurogenesis. However, this and other morphological characters remain of limited use for phylogenetic systematics owing to the lack of data from an appropriate outgroup. Here, we show that several embryonic characters are synapomorphies uniting the chelicerates and myriapods, as revealed by an outgroup comparison with the Onychophora or velvet worms. Our findings, thus provide, to our knowledge, first morphological/embryological support for the monophyly of the Paradoxopoda and suggest that the mandible might have evolved twice within the arthropods.
SoxS regulates the expression of the Salmonella enterica serovar Typhimurium ompW gene
Authors: Gil F. , Hernández-Lucas I. , Polanco R. , Pacheco N. , Collao B. , Villarreal J. M. , Nardocci G. , Calva E. , Saavedra C. P.
Published: July 2009

OmpW ofSalmonella entericaserovar Typhimurium has been described as a minor porin involved in osmoregulation, and is also affected by environmental conditions. Biochemical and genetic evidence from our laboratory indicates that OmpW is involved in efflux of and resistance towards paraquat (PQ), and its expression has been shown to be activated in response to oxidative stress. In this study we have exploredompWexpression in response to PQ. Primer extension and transcriptional fusions showed that its expression was induced in the presence of PQ.In silicoanalyses suggested a putative binding site for the SoxS transcriptional factor at theompWregulatory region. Electrophoretic mobility shift assays (EMSAs) and footprinting experiments showed that SoxS binds at a region that starts close to −54 and ends at about −197 upstream of the transcription start site. Transcriptional fusions support the relevance of this region inompWactivation. The SoxS site is in the forward orientation and its location suggests that theompWgene has a class I SoxS-dependent promoter.
A novel carotenoid 1,2-hydratase (CruF) from two species of the non-photosynthetic bacterium Deinococcus
Authors: Sun Zongtao , Shen Shaochuan , Wang Chao , Wang Hu , Hu Yaping , Jiao Jiandong , Ma Tingting , Tian Bing , Hua Yuejin
Published: July 2009

A novel carotenoid 1,2-hydratase (CruF) responsible for the C-1′,2′ hydration ofγ-carotene was identified in the non-photosynthetic bacteriaDeinococcus radioduransR1 andDeinococcus geothermalisDSM 11300. Gene expression and disruption experiments demonstrated thatdr0091anddgeo2309encode CruF inD. radioduransandD. geothermalis, respectively. Their homologues were also found in the genomes of cyanobacteria, and exhibited little homology to the hydroxyneurosporene synthase (CrtC) proteins found mainly in photosynthetic bacteria. Phylogenetic analysis showed that CruF homologues form a separate family, which is evolutionarily distant from the known CrtC family.
Unexpected extracellular and intracellular sulfur species during growth of Allochromatium vinosum with reduced sulfur compounds
Authors: Franz Bettina , Gehrke Thomas , Lichtenberg Henning , Hormes Josef , Dahl Christiane , Prange Alexander
Published: July 2009

Before its uptake and oxidation by purple sulfur bacteria, elemental sulfur probably first has to be mobilized. To obtain more insight into this mobilization process in the phototrophic purple sulfur bacterium Allochromatium vinosum, we used HPLC analysis and X-ray absorption near-edge structure (XANES) spectroscopy for the detection and identification of sulfur compounds in culture supernatants and bacterial cells. We intended to identify soluble sulfur compounds that specifically occur during growth on elemental sulfur, and therefore compared spectra of cultures grown on sulfur with those of cultures grown on sulfide or thiosulfate. While various unexpected oxidized organic sulfur species (sulfones, C–SO2–C, and sulfonates, ) were observed via XANES spectroscopy in the supernatants, we obtained evidence for the presence of monosulfane sulfonic acids inside the bacterial cells by HPLC analysis. The concentrations of the latter compounds showed a tight correlation with the content of intracellular sulfur, reaching their maximum when sulfur began to be oxidized. None of the detected sulfur compounds appeared to be a specific soluble intermediate or product of elemental sulfur mobilization. It therefore seems unlikely that mobilization of elemental sulfur by purple sulfur bacteria involves excretion of soluble sulfur-containing substances that would be able to act on substrate distant from the cells.
Interaction of triosephosphate isomerase from the cell surface of Staphylococcus aureus and α-(1→3)-mannooligosaccharides derived from glucuronoxylomannan of Cryptococcus neoformans
Authors: Furuya Hiromi , Ikeda Reiko
Published: July 2009

The glycolytic enzyme triosephosphate isomerase (TPI; EC 5.3.1.1) of Staphylococcus aureus is a candidate adhesion molecule for the interaction between the bacterium and the fungal pathogen Cryptococcus neoformans. TPI may recognize the mannan backbone of glucuronoxylomannan (GXM) of C. neoformans. We purified TPI from extracts of S. aureus surface proteins to investigate its binding by surface plasmon resonance analysis. The immobilized TPI reacted with GXM in a dose-dependent manner. Furthermore, the interactions between staphylococcal TPI and α-(1→3)-mannooligosaccharides derived from GXM were examined. The oligosaccharides exhibited binding with TPI; however, monomeric mannose did not. Differences in the slopes of the sensorgrams were observed between oligosaccharides with an even number of residues versus those with an odd number. A heterogeneous ligand-parallel reaction model revealed the existence of at least two binding sites on TPI. The enzymic activities of TPI were inhibited in a dose-dependent manner by α-(1→3)-mannooligosaccharides larger than triose. The binding of TPI and α-(1→3)-mannotriose near the substrate-binding site was predicted in silico (AutoDock 3.05). An oligosaccharide of size equal to or greater than triose could bind to the site, affecting enzymic activities. Moreover, affinities were indicated, especially for biose and tetraose, to another binding pocket, which would not affect enzymic activity. These data suggest a novel role for TPI, in addition to glycolysis, on the surface of S. aureus.
Resistance to peroxynitrite in Neisseria gonorrhoeae
Authors: Barth Kenneth R. , Isabella Vincent M. , Wright Lori F. , Clark Virginia L.
Published: July 2009

Neisseria gonorrhoeaeencodes a number of important genes that aid in survival during times of oxidative stress. The same immune cells capable of oxygen-dependent killing mechanisms also have the capacity to generate reactive nitrogen species (RNS) that may function antimicrobially. F62 and eight additional gonococcal strains displayed a high level of resistance to peroxynitrite, whileNeisseria meningitidisandEscherichia colishowed a four- to seven-log and a four-log decrease in viability, respectively. Mutation of gonococcal orthologues that are known or suspected to be involved in RNS defence in other bacteria (ahpC, dnrNandmsrA) resulted in no loss of viability, suggesting thatN. gonorrhoeaehas a novel mechanism of resistance to peroxynitrite. Whole-cell extracts of F62 prevented the oxidation of dihydrorhodamine, and decomposition of peroxynitrite was not dependent onahpC, dnrNormsrA. F62 grown in co-culture withE. colistrain DH10B was shown to protectE. coliviability 10-fold. Also, peroxynitrite treatment of F62 did not result in accumulation of nitrated proteins, suggesting that an active peroxynitrite reductase is responsible for peroxynitrite decomposition rather than a protein sink for amino acid modification.
Functional analyses of pilin-like proteins from Francisella tularensis: complementation of type IV pilus phenotypes in Neisseria gonorrhoeae
Authors: Salomonsson Emelie , Forsberg Åke , Roos Norbert , Holz Claudia , Maier Berenike , Koomey Michael , Winther-Larsen Hanne C.
Published: July 2009

Accumulating evidence from a number of studies strongly suggests that proteins orthologous to those involved in type IV pili (Tfp) assembly and function are required for Francisella pathogenicity. However, the molecular mechanisms by which the components exert their influence on virulence remain poorly understood. Owing to the conservation and promiscuity of Tfp biogenesis machineries, expression of Tfp pilins in heterologous species has been used successfully to analyse organelle structure–function relationships. In this study we expressed a number of Francisella pilin genes in the Tfp-expressing pathogen Neisseria gonorrhoeae lacking its endogenous pilin subunit. Two gene products, the orthologous PilA proteins from Francisella tularensis subspecies tularensis and novicida, were capable of restoring the expression of Tfp-like appendages that were shown to be dependent upon the neisserial Tfp biogenesis machinery for surface localization. Expression of Francisella PilA pilins also partially restored competence for natural transformation in N. gonorrhoeae. This phenotype was not complemented by expression of the PulG and XcpT proteins, which are equivalent components of the related type II protein secretion system. Taken together, these findings provide compelling, although indirect, evidence of the potential for Francisella PilA proteins to express functional Tfp.
Metabolic control through ornithine and uracil of epithelial cell invasion by Shigella flexneri
Authors: Durand Jérôme M. B. , Björk Glenn R.
Published: July 2009

This paper shows that compounds in defined growth media strongly influence the expression of the effectors of virulence in the human invasive pathogenShigella flexneri. Ornithine in conjunction with uracil reduces the haemolytic ability of wild-type cultures more than 20-fold and the expression of the type III secretion system more than 8-fold, as monitored by anmxiC : : lacZtranscriptional reporter.mxiCgene expression is further decreased by the presence of methionine or branched-chain amino acids (15-fold or 25-fold at least, respectively). Lysine and a few other aminated metabolites (cadaverine, homoserine and diaminopimelate) counteract the ornithine-mediated inhibition of haemolytic activity and of the expression of a transcriptional activatorvirFreporter. The complete abolition of invasion of HeLa cells by wild-type bacteria by ornithine, uracil, methionine or branched-chain amino acids establishes that these metabolites are powerful effectors of virulence. These findings provide a direct connection between metabolism and virulence inS. flexneri. The inhibitory potential exhibited by the nutritional environment is stronger than temperature, the classical environmental effector of virulence. The implications and practical application of this finding in prophylaxis and treatment of shigellosis are discussed.
Intercistronic heterogeneity of the 16S–23S rRNA spacer region among Pseudomonas strains isolated from subterranean seeds of hog peanut (Amphicarpa bracteata)
Authors: Tambong J. T. , Xu R. , Bromfield E. S. P.
Published: July 2009

Intercistronic heterogeneity of the 16S–23S rRNA internal transcribed spacer regions (ITS1) was investigated in 29 strains of fluorescent pseudomonads isolated from subterranean seeds ofAmphicarpa bracteata(hog peanut). PCR amplification of the ITS1 region generated one or two products from the strains. Sequence analysis of the amplified fragments revealed an ITS1 fragment of about 517 bp that contained genes for tRNAIleand tRNAAlain all 29 strains; an additional smaller ITS1 of 279 bp without tRNA features was detected in 15 of the strains. The length difference appeared to be due to deletions of several nucleotide blocks between the 70 bp and 359 bp positions of the alignment. The end of the deletions in the variant ITS1 type coincided with the start of antiterminator box A, which is homologous to box A of other bacteria. Phylogenetic analyses using the neighbour-joining algorithm revealed two major phylogenetic clusters, one for each of the ITS1 types. Using a single specific primer set and the DNA-intercalating dye SYBR Green I for real-time PCR and melting-curve analysis produced highly informative curves with one or two recognizable melting peaks that readily distinguished between the two ITS1 types in pure cultures. The assay was used to confirm the presence of the variant ITS1 type in thePseudomonascommunity in total DNA from root-zone soil and seed coats of hog peanut. Heterogeneity of the ITS1 region between species has potential for studying molecular systematics and population genetics of the genusPseudomonas, but the presence of non-identical rRNA operons within a genome may pose problems.
Localization of the equine IgG-binding domain in the fibrinogen-binding protein (FgBP) of Streptococcus equi subsp. equi
Authors: Meehan Mary , Lewis Melanie J. , Byrne Caroline , O'Hare David , Woof Jenny M. , Owen Peter
Published: July 2009

Fibrinogen-binding protein (FgBP, also termed SeM) is a cell-wall-associated anti-phagocytic M-like protein of the equine pathogenStreptococcus equisubsp.equi, and binds fibrinogen (Fg) and IgG. FgBP binds Fg avidly through residues located at the extreme N terminus of the molecule, whereas the IgG-binding site is more centrally located between the A and B repeats. FgBP binds equine IgG4 and IgG7 subclasses through interaction with the CH2–CH3 interdomain region of IgG-Fc, and possesses overlapping Fc-binding sites with protein A and protein G. In this study, FgBP truncates containing defined internal deletions were used to identify a stretch of 14 aa (residues 335–348) critical for IgG binding. Protein chimeras consisting of the non-IgG-bindingα-helical coiled-coil M5 protein fused to FgBP sequences were used to identify a minimal equine IgG-binding domain consisting of residues 329–360. Competition ELISA tests suggested that IgG does not compromise Fg binding and vice versa.
Enhanced citric acid biosynthesis in Pseudomonas fluorescens ATCC 13525 by overexpression of the Escherichia coli citrate synthase gene
Authors: Buch Aditi D. , Archana G. , Kumar G. Naresh
Published: July 2009

Citric acid secretion by fluorescent pseudomonads has a distinct significance in microbial phosphate solubilization. The role of citrate synthase in citric acid biosynthesis and glucose catabolism in pseudomonads was investigated by overexpressing theEscherichia colicitrate synthase (gltA) gene inPseudomonas fluorescensATCC 13525. The resultant ∼2-fold increase in citrate synthase activity in thegltA-overexpressing strainPf(pAB7) enhanced the intracellular and extracellular citric acid yields during the stationary phase, by about 2- and 26-fold, respectively, as compared to the control, without affecting the growth rate, glucose depletion rate or biomass yield. Decreased glucose consumption was paralleled by increased gluconic acid production due to an increase in glucose dehydrogenase activity. While the extracellular acetic acid yield increased inPf(pAB7), pyruvic acid secretion decreased, correlating with an increase in pyruvate carboxylase activity and suggesting an increased demand for the anabolic precursor oxaloacetate. Activities of two other key enzymes, glucose-6-phosphate dehydrogenase and isocitrate dehydrogenase, remained unaltered, and the contribution of phosphoenolpyruvate carboxylase and isocitrate lyase to glucose catabolism was negligible. StrainPf(pAB7) demonstrated an enhanced phosphate-solubilizing ability compared to the control. Co-expression of theSynechococcus elongatusPCC 6301 phosphoenolpyruvate carboxylase andE. coli gltAgenes inP. fluorescensATCC 13525, so as to supplement oxaloacetate for citrate biosynthesis, neither significantly affected citrate biosynthesis nor caused any change in the other physiological and biochemical parameters measured, despite approximately 1.3- and 5-fold increases in citrate synthase and phosphoenolpyruvate carboxylase activities, respectively. Thus, our results demonstrate that citrate synthase is rate-limiting in enhancing citrate biosynthesis inP. fluorescensATCC 13525. Significantly low extracellular citrate levels as compared to the intracellular levels inPf(pAB7) suggested a probable limitation of efficient citrate transport.
Pivotal role of the Francisella tularensis heat-shock sigma factor RpoH
Authors: Grall Nathalie , Livny Jonathan , Waldor Matthew , Barel Monique , Charbit Alain , Meibom Karin L.
Published: July 2009

Francisella tularensis is a highly infectious pathogen that infects animals and humans to cause the disease tularemia. The primary targets of this bacterium are macrophages, in which it replicates in the cytoplasm after escaping the initial phagosomal compartment. The ability to replicate within macrophages relies on the tightly regulated expression of a series of genes. One of the most commonly used means of coordinating the regulation of multiple genes in bacteria consists of the association of dedicated alternative sigma factors with the core of the RNA polymerase (RNAP). In silico analysis of the F. tularensis LVS genome led us to identify, in addition to the genes encoding the RNAP core (comprising the α1, α2, β, β′ and ω subunits), one gene (designated rpoD) encoding the major sigma factor σ
70, and a unique gene (FTL_0851) encoding a putative alternative sigma factor homologue of the σ
32 heat-shock family (designated rpoH). Hence, F. tularensis represents one of the minority of bacterial species that possess only one or no alternative sigma factor in addition to the main factor σ
70. In the present work, we show that FTL_0851 encodes a genuine σ
32 factor. Transcriptomic analyses of the F. tularensis LVS heat-stress response allowed the identification of a series of orthologues of known heat-shock genes (including those for Hsp40, GroEL, GroES, DnaK, DnaJ, GrpE, ClpB and ClpP) and a number of genes implicated in Francisella virulence. A bioinformatic analysis was used to identify genes preceded by a putative σ
32-binding site, revealing both similarities to and differences from RpoH-mediated gene expression in Escherichia coli. Our results suggest that RpoH is an essential protein of F. tularensis, and positively regulates a subset of genes involved in the heat-shock response.