bluuebunny/crossref_metadata_2025 huggingface dataset:
FGF-1 induces ATP release from spinal astrocytes in culture and opens pannexin and connexin hemichannels
Authors: Garré Juan M. , Retamal Mauricio A. , Cassina Patricia , Barbeito Luis , Bukauskas Feliksas F. , Sáez Juan C. , Bennett Michael V. L. , Abudara Verónica
Published: December 2010

Spinal astrocytes are coupled by connexin (Cx) gap junctions and express pannexin 1 (Px1) and purinergic receptors. Fibroblast growth factor 1 (FGF-1), which is released in spinal cord injury, activated spinal astrocytes in culture, induced secretion of ATP, and permeabilized them to relatively large fluorescent tracers [ethidium (Etd) and lucifer yellow (LY)] through “hemichannels” (HCs). HCs can be formed by connexins or pannexins; they can open to extracellular space or can form gap junction (GJ) channels, one HC from each cell. (Pannexins may not form gap junctions in mammalian tissues, but they do in invertebrates). HC types were differentiated pharmacologically and by Px1 knockdown with siRNA and by use of astrocytes from Cx43 knockout mice. Permeabilization was reduced by apyrase (APY), an ATPase, and by P2X
7
receptor antagonists, implicating secretion of ATP and autocrine and/or paracrine action. Increased permeability of cells exposed to FGF-1 or ATP for 2 h was mediated largely by Px1 HCs activated by P2X
7
receptors. After a 7-h treatment, the permeability was mediated by both Cx43 and Px1 HCs. FGF-1 also caused reduction in gap junctional communication. Botulinum neurotoxin A, a blocker of vesicular release, reduced permeabilization when given 30 min before FGF-1 application, but not when given 1 h after FGF-1. We infer that ATP is initially released from vesicles and then it mediates continued release by action on P2X
7
receptors and opening of HCs. These changes in HCs and gap junction channels may promote inflammation and deprive neurons of astrocyte-mediated protection in spinal cord trauma and neurodegenerative disease.
Do Not Give Up Hope for a Climate Agreement
Authors: Karp Larry
Published: December 2010

The failure of the 2009 Copenhagen climate negotiations does not encourage optimism about reaching a meaningful climate agreement. This paper describes a proposal for the next climate agreement that entitles countries to discharge their treaty obligations by paying a ‘fine’. This escape clause provides cost insurance, simplifies the problem of enforcing compliance, and increases incentives to participate in the agreement. The proposal requires that developed countries commit to emissions reductions, maintained by means of cap and trade rather than carbon taxes. A Central Bank maintains stability of carbon prices by defending a price ceiling and floor.
Green Regulations in California and Sweden
Authors: Berck Peter , Brännlund Runar , Berck Cyndi Spindell
Published: December 2010

California and Sweden are both leaders in green regulations and actions. In both there is a substantial political base for environmental regulation, yet the path to regulation in these two political entities is quite different. California emphasizes command and control regulations while Sweden makes heavy use of taxes. We show that both underlying economic factors and the constraints of the larger systems in which these economies are embedded contribute to their choice of control methods.
Mugsy: fast multiple alignment of closely related whole genomes
Authors: Angiuoli Samuel V. , Salzberg Steven L.
Published: December 2010

Motivation: The relative ease and low cost of current generation sequencing technologies has led to a dramatic increase in the number of sequenced genomes for species across the tree of life. This increasing volume of data requires tools that can quickly compare multiple whole-genome sequences, millions of base pairs in length, to aid in the study of populations, pan-genomes, and genome evolution.

Results: We present a new multiple alignment tool for whole genomes named Mugsy. Mugsy is computationally efficient and can align 31 Streptococcus pneumoniae genomes in less than 2 hours producing alignments that compare favorably to other tools. Mugsy is also the fastest program evaluated for the multiple alignment of assembled human chromosome sequences from four individuals. Mugsy does not require a reference sequence, can align mixtures of assembled draft and completed genome data, and is robust in identifying a rich complement of genetic variation including duplications, rearrangements, and large-scale gain and loss of sequence.

Availability: Mugsy is free, open-source software available from http://mugsy.sf.net.

Contact:  angiuoli@cs.umd.edu

Supplementary information:  Supplementary data are available at Bioinformatics online.
Multinational Enterprise and Government Controls on Outward Foreign Direct Investment in the United States and the United Kingdom in the 1960s
Authors: Rollings Neil
Published: December 2010

The 1960s have been seen as critical in the development of multinational enterprise by social scientists. Business historians have provided a context to these developments but are only now beginning to consider the 1960s in the light of access to archival material. This article analyses the nature and extent of the tension between multinational enterprise and U.S. and U.K. national governments in the 1960s caused by the tightening of controls on FDI introduced to resolve balance of payments problems. A comparative approach is adopted to illustrate that these tensions were not restricted to the U.S. The tension revolved around different conceptions of business investment decision-making and of the contribution of multinationals to the national economy. It is also suggested that the impact of these measures was at the micro-level and this highlights the value of business history approaches.
Self‐care management of heart failure: practical recommendations from the Patient Care Committee of the Heart Failure Association of the European Society of Cardiology
Authors: Lainscak Mitja , Blue Lynda , Clark Andrew L. , Dahlström Ulf , Dickstein Kenneth , Ekman Inger , McDonagh Theresa , McMurray John J. , Ryder Mary , Stewart Simon , Strömberg Anna , Jaarsma Tiny
Published: December 2010

Guidelines on heart failure (HF) stress the importance of lifestyle advice, although there is little evidence that such recommendations improve symptoms or prognosis. Patients experience symptoms of different intensities which impair their daily activities and reduce the quality‐of‐life. To cope with their clinical condition, many patients seek advice about lifestyle and self‐management strategies when in contact with medical care providers, particularly specialized HF services. Self‐care management is an important part of HF treatment, thus health professionals working with patients with HF have recognized the need for more specific recommendations on lifestyle advice. The present paper summarizes the available evidence, promotes self‐care management, and aims to provide practical advice for health professionals delivering care to HF patients. It also defines avenues of research to optimize self‐care strategies in a number of key areas to derive further benefits.
3,423 Emergency Tracheal Intubations at a University Hospital
Authors: Martin Lizabeth D. , Mhyre Jill M. , Shanks Amy M. , Tremper Kevin K. , Kheterpal Sachin
Published: December 2010

Background

There are limited outcome data regarding emergent nonoperative intubation. The current study was undertaken with a large observational dataset to evaluate the incidence of difficult intubation and complication rates and to determine predictors of complications in this setting.

Methods

Adult nonoperating room emergent intubations at our tertiary care institution from December 5, 2001 to July 6, 2009 were reviewed. Prospectively defined data points included time of day, location, attending physician presence, number of attempts, direct laryngoscopy view, adjuvant use, medications, and complications. At our institution, a senior resident with at least 24 months of anesthesia training is the first responder for all emergent airway requests. The primary outcome was a composite airway complication variable that included aspiration, esophageal intubation, dental injury, or pneumothorax.

Results

A total of 3,423 emergent nonoperating room airway management cases were identified. The incidence of difficult intubation was 10.3%. Complications occurred in 4.2%: aspiration, 2.8%; esophageal intubation, 1.3%; dental injury, 0.2%; and pneumothorax, 0.1%. A bougie introducer was used in 12.4% of cases. Among 2,284 intubations performed by residents, independent predictors of the composite complication outcome were as follows: three or more intubation attempts (odds ratio, 6.7; 95% CI, 3.2-14.2), grade III or IV view (odds ratio, 1.9; 95% CI, 1.1-3.5), general care floor location (odds ratio, 1.9; 95% CI, 1.2-3.0), and emergency department location (odds ratio, 4.7; 95% CI, 1.1-20.4).

Conclusions

During emergent nonoperative intubation, specific clinical situations are associated with an increased risk of airway complication and may provide a starting point for allocation of experienced first responders.
Routine Clinical Practice Effectiveness of the Glidescope in Difficult Airway Management
Authors: Aziz Michael F. , Healy David , Kheterpal Sachin , Fu Rongwei F. , Dillman Dawn , Brambrink Ansgar M.
Published: December 2010

Introduction

The Glidescope video laryngoscope has been shown to be a useful tool to improve laryngeal view. However, its role in the daily routine of airway management remains poorly characterized.

Methods

This investigation evaluated the use of the Glidescope at two academic medical centers. Electronic records from 71,570 intubations were reviewed, and 2,004 cases were identified where the Glidescope was used for airway management. We analyzed the success rate of Glidescope intubation in various intubation scenarios. In addition, the incidence and character of complications associated with Glidescope use were recorded. Predictors of Glidescope intubation failure were determined using a logistic regression analysis.

Results

Overall success for Glidescope intubation was 97% (1,944 of 2,004). As a primary technique, success was 98% (1,712 of 1,755), whereas success in patients with predictors of difficult direct laryngoscopy was 96% (1,377 of 1,428). Success for Glidescope intubation after failed direct laryngoscopy was 94% (224 of 239). Complications were noticed in 1% (21 of 2,004) of patients and mostly involved minor soft tissue injuries, but major complications, such as dental, pharyngeal, tracheal, or laryngeal injury, occurred in 0.3% (6 of 2,004) of patients. The strongest predictor of Glidescope failure was altered neck anatomy with presence of a surgical scar, radiation changes, or mass.

Conclusion

These data demonstrate a high success rate of Glidescope intubation in both primary airway management and rescue-failed direct laryngoscopy. However, Glidescope intubation is not always successful and certain predictors of failure can be identified. Providers should maintain their competency with alternate methods of intubation, especially for patients with neck pathology.
Clostridium chromiireducens sp. nov., isolated from Cr(VI)-contaminated soil
Authors: Inglett K. S. , Bae H. S. , Aldrich H. C. , Hatfield K. , Ogram A. V.
Published: December 2010

A Cr(VI)-resistant, Gram-positive, spore-forming, obligate anaerobe, designated GCAF-1T, was isolated from chromium-contaminated soil by its ability to reduce Cr(VI) in low concentrations. Mixed acid fermentation during growth on glucose resulted in accumulation of acetate, butyrate, formate and lactate. Morphological studies indicated the presence of peritrichous flagella, pili and an S-layer. The major cellular fatty acids (>5 %) were C16 : 0, C14 : 0, summed feature 3 (comprising iso-C15 : 0 2-OH and/or C16 : 1ω7c), C18 : 1ω7c, C16 : 1ω9c, summed feature 4 (comprising iso-C17 : 1 I and/or anteiso-C17 : 1 B) and C18 : 1ω9c. The DNA G+C content of strain GCAF-1T was 30.7 mol%. Phylogenetic interference indicated that strain GCAF-1T clustered with group I of the genus Clostridium. Of strains within this cluster, strain GCAF-1T shared the highest 16S rRNA gene sequence similarities (98.1–98.9 %) with Clostridium beijerinckii DSM 791T, C. saccharobutylicum NCP 262T, C. saccharoperbutylacetonicum N1-4T, C. puniceum DSM 2619T and C. roseum DSM 51T. However, strain GCAF-1T could be clearly distinguished from its closest phylogenetic neighbours by low levels of DNA–DNA relatedness (<50 %) and some phenotypic features. Based on the evidence presented here, strain GCAF-1T ( = DSM 23318T  = KCTC 5935T) represents a novel species of the genus Clostridium, for which the name Clostridium chromiireducens sp. nov. is proposed.
Gramella gaetbulicola sp. nov., a member of the family Flavobacteriaceae isolated from foreshore soil
Authors: Cho Sung-Heun , Chae Song-Hee , Cho Myoungho , Kim Tae-Ui , Choi Seri , Han Ji-Hye , Kim Young Tae , Joung Yochan , Joh Kiseong , Nedashkovskaya Olga I. , Kim Seung Bum
Published: December 2010

A Gram-reaction-negative, yellow-pigmented, gliding, rod-shaped, aerobic bacterium (RA5-111T) was isolated from foreshore soil. The taxonomic status of the novel isolate was determined using a polyphasic approach. On the basis of 16S rRNA gene sequence similarities, strain RA5-111T could be assigned to the genus Gramella, with sequence similarities of 97.7, 97.3 and 96.2 % to the type strains of Gramella echinicola, Gramella portivictoriae and Gramella marina, respectively. Chemotaxonomic and phenotypic characteristics also supported the affiliation of strain RA5-111T with the genus Gramella. The genomic DNA G+C content was 39.1 mol%. The isolate contained MK-6 as the predominant menaquinone, iso-C15 : 0, iso-C17 : 0 3-OH and a summed feature (iso-C15 : 0 2-OH and/or C16 : 1ω7c) as major fatty acids, and phosphatidylethanolamine and unknown phospholipids as the polar lipids. DNA–DNA relatedness, phenotypic, genotypic and chemotaxonomic data clearly indicate that the isolate represents a novel species of the genus Gramella, for which the name Gramella gaetbulicola sp. nov. is proposed. The type strain is RA5-111T ( = KCTC 23022T  = JCM 16528T  = NBRC 106272T).
Streptomyces sundarbansensis sp. nov., an actinomycete that produces 2-allyloxyphenol
Authors: Arumugam Meyyappan , Mitra Anindita , Pramanik Arnab , Saha Malay , Gachhui Ratan , Mukherjee Joydeep
Published: December 2010

A novel actinomycete producing 2-allyloxyphenol, designated strain MS1/7T, was isolated from sediments of the Sundarbans mangrove forest, India. Growth on International Streptomyces Project (ISP) media 2, 3, 4, 5 and 7 produced olive green to grey aerial hyphae that carried smooth-surfaced spores in a flexuous (Rectiflexibiles) arrangement. The strain contained ll-diaminopimelic acid, but no diagnostic sugars in whole-cell hydrolysates. Hexa-, octa- and a minor amount of tetra-hydrogenated menaquinones with nine isoprene units [MK-9 (H4, H6, H8 and H10)] were present as isoprene analogues. Diagnostic phospholipids were phosphatidylethanolamine and diphosphatidylglycerol. The predominant fatty acids were anteiso-C15 : 0 (34.80 %), iso-C16 : 0 (16.45 %), C16 (10.53 %) and anteiso-C17 : 0 (10.92 %). The strain showed greater than 99 % 16S rRNA gene sequence similarity to the type strains of several recognized species of the genus Streptomyces, but in the phylogenetic tree based on 16S rRNA gene sequences it formed a distinct phyletic line and demonstrated closest relationships to viomycin-producers (Streptomyces californicus NRRL B-1221T, Streptomyces floridae MTCC 2534T and Streptomyces puniceus NRRL B-2895T). However, strain MS1/7T could be distinguished from these and other closely related species based on low levels of DNA–DNA relatedness (<44 %) and disparate physiological features, principally amino acid utilization and growth in NaCl. Strain MS1/7T is therefore suggested to represent a novel species of the genus Streptomyces, for which the name Streptomyces sundarbansensis sp. nov. is proposed. The type strain is MS1/7T ( = MTCC 10621T = DSM 42019T).
Actinopolymorpha pittospori sp. nov., an endophyte isolated from surface-sterilized leaves of an apricot tree (Pittosporum phylliraeoides)
Authors: Kaewkla Onuma , Franco Christopher M. M.
Published: December 2010

A member of the genus Actinopolymorpha, designated PIP 143T, was isolated from the leaves of an Australian native apricot tree (Pittosporum phylliraeoides). The isolate was a Gram-reaction-positive, aerobic actinobacterium, with a well-developed substrate mycelium that fragmented into small rods. Phylogenetic evaluation based on 16S rRNA gene sequences placed the isolate in the family Nocardioidaceae. Strain PIP 143T was most closely related to Actinopolymorpha cephalotaxi I06-2230T (98.7 %) and Actinopolymorpha rutila YIM 45725T (98.1 %). Chemotaxonomic data, including cell-wall components, menaquinones and fatty acids, confirmed the affiliation of strain PIP 143T to the genus Actinopolymorpha. Phylogenetic analysis and physiological and biochemical studies, in combination with DNA–DNA hybridization studies, allowed the differentiation of strain PIP 143T from its closest phylogenetic neighbours with validly published names. Therefore, a novel species is proposed, with the name Actinopolymorpha pittospori sp. nov. The type strain is PIP 143T ( = DSM 45354T  = ACM 5288T  = NRRL B-24810T).
Root-Specific Reduction of Cytokinin Causes Enhanced Root Growth, Drought Tolerance, and Leaf Mineral Enrichment inArabidopsisand Tobacco
Authors: Werner Tomáš , Nehnevajova Erika , Köllmer Ireen , Novák Ondřej , Strnad Miroslav , Krämer Ute , Schmülling Thomas
Published: December 2010

Optimizing root system architecture can overcome yield limitations in crop plants caused by water or nutrient shortages. Classic breeding approaches are difficult because the trait is governed by many genes and is difficult to score. We generated transgenic Arabidopsis thaliana and tobacco (Nicotiana tabacum) plants with enhanced root-specific degradation of the hormone cytokinin, a negative regulator of root growth. These transgenic plants form a larger root system, whereas growth and development of the shoot are similar. Elongation of the primary root, root branching, and root biomass formation were increased by up to 60% in transgenic lines, increasing the root-to-shoot ratio. We thus demonstrated that a single dominant gene could regulate a complex trait, root growth. Moreover, we showed that cytokinin regulates root growth in a largely organ-autonomous fashion that is consistent with its dual role as a hormone with both paracrine and long-distance activities. Transgenic plants had a higher survival rate after severe drought treatment. The accumulation of several elements, including S, P, Mn, Mg, Zn, as well as Cd from a contaminated soil, was significantly increased in shoots. Under conditions of sulfur or magnesium deficiency, leaf chlorophyll content was less affected in transgenic plants, demonstrating the physiological relevance of shoot element accumulation. Our approach might contribute to improve drought tolerance, nutrient efficiency, and nutrient content of crop plants.
Inhibiting the PI3K/Akt pathway reversed progestin resistance in endometrial cancer
Authors: Gu Chao , Zhang Zhenbo , Yu Yinhua , Liu Yingtao , Zhao Fengdi , Yin Lianhua , Feng Youji , Chen Xiaojun
Published: December 2010

Progestin resistance is the main obstacle to successful conservative therapy in young endometrial cancer patients. To investigate the molecular events that lead to progestin resistance and to find a possible way to reverse progestin resistance in endometrial cancer, we established a progestin‐resistant Ishikawa cell line by long‐term progestin treatment to downregulate progesterone receptor (PR) expression. Both medoxyprogesterone acetate (MPA) and LY294002, a phosphatidylinositol 3‐kinase (PI3K) inhibitor, were assayed for their effects on the proliferation of progestin‐sensitive and progestin‐resistant cancer cells, respectively. The MPA inhibited the PI3K/Akt pathway and suppressed cell proliferation in progestin‐sensitive Ishikawa cells, but activated the PI3K/Akt pathway and had no effect on cell proliferation in progestin‐resistant Ishikawa cells or HEC‐1A cells. Inhibiting the PI3K/Akt pathway by LY294002 upregulated PR expression and diminished cell growth, especially in progestin‐resistant endometrial cancer cells. In vivo endometrial cancer xenograft studies in nude mice also showed that inhibiting the PI3K/Akt pathway reversed progestin resistance in endometrial cancer. Our results indicate that activation of the PI3K/Akt pathway by progestin without PR mediation plays an important role in progestin resistance to endometrial cancer cells. In addition, inhibiting the PI3K/Akt pathway might reverse progestin resistance in endometrial cancer. (Cancer Sci 2011; 102: 557–564)
Among Developmental Regulators, StuA but Not BrlA Is Essential for Penicillin V Production in
Penicillium chrysogenum
Authors: Sigl Claudia , Haas Hubertus , Specht Thomas , Pfaller Kristian , Kürnsteiner Hubert , Zadra Ivo
Published: December 2010

In filamentous fungi, secondary metabolism is often linked with developmental processes such as conidiation. In this study we analyzed the link between secondary metabolism and conidiation in the main industrial producer of the β-lactam antibiotic penicillin, the ascomycete
Penicillium chrysogenum
. Therefore, we generated mutants defective in two central regulators of conidiation, the transcription factors BrlA and StuA. Inactivation of either
brlA
or
stuA
blocked conidiation and altered hyphal morphology during growth on solid media, as shown by light and scanning electron microscopy, but did not affect biomass production during liquid-submerged growth. Genome-wide transcriptional profiling identified a complex StuA- and BrlA-dependent regulatory network, including genes previously shown to be involved in development and secondary metabolism. Remarkably, inactivation of
stuA
, but not
brlA
, drastically downregulated expression of the penicillin biosynthetic gene cluster during solid and liquid-submerged growth. In agreement, penicillin V production was wild-type-like in
brlA
-deficient strains but 99% decreased in
stuA
-deficient strains during liquid-submerged growth, as shown by high-performance liquid chromatography (HPLC) analysis. Thus, among identified regulators of penicillin V production StuA has the most severe influence. Overexpression of
stuA
increased the transcript levels of
brlA
and
abaA
(another developmental regulator) and derepressed conidiation during liquid-submerged growth but did not affect penicillin V productivity. Taken together, these data demonstrate an intimate but not exclusive link between regulation of development and secondary metabolism in
P. chrysogenum.
Bacterial Killing by Dry Metallic Copper Surfaces
Authors: Santo Christophe Espírito , Lam Ee Wen , Elowsky Christian G. , Quaranta Davide , Domaille Dylan W. , Chang Christopher J. , Grass Gregor
Published: December 2010

Metallic copper surfaces rapidly and efficiently kill bacteria. Cells exposed to copper surfaces accumulated large amounts of copper ions, and this copper uptake was faster from dry copper than from moist copper. Cells suffered extensive membrane damage within minutes of exposure to dry copper. Further, cells removed from copper showed loss of cell integrity. Acute contact with metallic copper surfaces did not result in increased mutation rates or DNA lesions. These findings are important first steps for revealing the molecular sensitive targets in cells lethally challenged by exposure to copper surfaces and provide a scientific explanation for the use of copper surfaces as antimicrobial agents for supporting public hygiene.
Linking Low-Level Stable Isotope Fractionation to Expression of the Cytochrome P450 Monooxygenase-Encoding
ethB
Gene for Elucidation of Methyl
tert
-Butyl Ether Biodegradation in Aerated Treatment Pond Systems
Authors: Jechalke Sven , Rosell Mònica , Martínez-Lavanchy Paula M. , Pérez-Leiva Paola , Rohwerder Thore , Vogt Carsten , Richnow Hans H.
Published: December 2010

Multidimensional compound-specific stable isotope analysis (CSIA) was applied in combination with RNA-based molecular tools to characterize methyl tertiary (
tert
-) butyl ether (MTBE) degradation mechanisms occurring in biofilms in an aerated treatment pond used for remediation of MTBE-contaminated groundwater. The main pathway for MTBE oxidation was elucidated by linking the low-level stable isotope fractionation (mean carbon isotopic enrichment factor [ε
C
] of −0.37‰ ± 0.05‰ and no significant hydrogen isotopic enrichment factor [ε
H
]) observed in microcosm experiments to expression of the
ethB
gene encoding a cytochrome P450 monooxygenase able to catalyze the oxidation of MTBE in biofilm samples both from the microcosms and directly from the ponds. 16S rRNA-specific primers revealed the presence of a sequence 100% identical to that of
Methylibium petroleiphilum
PM1, a well-characterized MTBE degrader. However, neither expression of the
mdpA
genes encoding the alkane hydroxylase-like enzyme responsible for MTBE oxidation in this strain nor the related MTBE isotope fractionation pattern produced by PM1 could be detected, suggesting that this enzyme was not active in this system. Additionally, observed low inverse fractionation of carbon (ε
C
of +0.11‰ ± 0.03‰) and low fractionation of hydrogen (ε
H
of −5‰ ± 1‰) in laboratory experiments simulating MTBE stripping from an open surface water body suggest that the application of CSIA in field investigations to detect biodegradation may lead to false-negative results when volatilization effects coincide with the activity of low-fractionating enzymes. As shown in this study, complementary examination of expression of specific catabolic genes can be used as additional direct evidence for microbial degradation activity and may overcome this problem.
Quantification and Genotyping of Human Sapoviruses in the Llobregat River Catchment, Spain
Authors: Sano Daisuke , Pérez-Sautu Unai , Guix Susana , Pintó Rosa Maria , Miura Takayuki , Okabe Satoshi , Bosch Albert
Published: December 2010

Human sapoviruses (SaVs) were quantified and characterized in an 18-month survey conducted along the Llobregat river catchment area in Spain. Sample types included freshwater, untreated and treated wastewater, and drinking water. All genogroups were recovered, and a seasonal distribution was observed. This is the first report of SaV quantification and genotyping in the environment outside Japan.
Wide Host Range and Strong Lytic Activity ofStaphylococcusaureusLytic Phage Stau2
Authors: Hsieh Sue-Er , Lo Hsueh-Hsia , Chen Shui-Tu , Lee Mong-Chuan , Tseng Yi-Hsiung
Published: December 2010

In searching for an alternative antibacterial agent against multidrug-resistantStaphylococcusaureus, we have isolated and characterized a lytic staphylophage, Stau2. It possesses a double-stranded DNA genome estimated to be about 134.5 kb and a morphology resembling that of members of the familyMyoviridae. With an estimated latency period of 25 min and a burst size of 100 PFU/infected cell, propagation of Stau2 in liquid culture gave a lysate of ca. 6 × 1010PFU/ml. It was stable at pH 5 to 13 in normal saline at room temperature for at least 4 weeks and at −85°C for more than 2 years, while 1 × 109out of 2 × 1012PFU/ml retained infectivity after 36 months at 4°C. Stau2 could lyse 80% of theS.aureusisolates (164/205) obtained from hospitals in Taiwan, with complete lysis of most of the isolates tested within 3 h; however, it was anS.aureus-specific phage because no lytic infection could be found in the coagulase-negative staphylococci tested. Its host range amongS.aureusisolates was wider than that of polyvalent phage K (47%), which can also lyse many other staphylococcal species. Experiments with mice demonstrated that Stau2 could provide 100% protection from lethal infection when a multiplicity of infection of 10 was administered immediately after a challenge withS.aureusS23. Considering these results, Stau2 could be considered at least as a candidate for topical phage therapy or an additive in the food industry.
New Anaerobic, Ammonium-Oxidizing Community Enriched from Peat Soil
Authors: Hu Bao-lan , Rush Darci , van der Biezen Erwin , Zheng Ping , van Mullekom Mark , Schouten Stefan , Sinninghe Damsté Jaap S. , Smolders Alfons J. P. , Jetten Mike S. M. , Kartal Boran
Published: December 2010

Anaerobic ammonium-oxidizing (anammox) bacteria have been recognized as an important sink for fixed nitrogen and are detected in many natural environments. However, their presence in terrestrial ecosystems has long been overlooked, and their contribution to the nitrogen cycling in natural and agricultural soils is currently unknown. Here we describe the enrichment and characterization of anammox bacteria from a nitrogen-loaded peat soil. After 8 months of incubation with the natural surface water of the sampling site and increasing ammonium and nitrite concentrations, anammox cells constituted 40 to 50% of the enrichment culture. The two dominant anammox phylotypes were affiliated with “
Candidatus
Jettenia asiatica” and “
Candidatus
Brocadia fulgida.” The enrichment culture converted NH
4
+
and NO
2
−
to N
2
with the previously reported stoichiometry (1:1.27) and had a maximum specific anaerobic ammonium oxidation rate of 0.94 mmol NH
4
+
·g (dry weight)
−1
·h
−1
at pH 7.1 and 32°C. The diagnostic anammox-specific lipids were detected at a concentration of 650 ng·g (dry weight)
−1
, and pentyl-[3]-ladderane was the most abundant ladderane lipid.
Using the Systematic Review Methodology To Evaluate Factors That Influence the Persistence of Influenza Virus in Environmental Matrices
Authors: Irwin C. K. , Yoon K. J. , Wang C. , Hoff S. J. , Zimmerman J. J. , Denagamage T. , O'Connor A. M.
Published: December 2010

Understanding factors that influence persistence of influenza virus in an environment without host animals is critical to appropriate decision-making for issues such as quarantine downtimes, setback distances, and eradication programs in livestock production systems. This systematic review identifies literature describing persistence of influenza virus in environmental samples, i.e., air, water, soil, feces, and fomites. An electronic search of PubMed, CAB, AGRICOLA, Biosis, and Compendex was performed, and citation relevance was determined according to the aim of the review. Quality assessment of relevant studies was performed using criteria from experts in virology, disease ecology, and environmental science. A total of 9,760 abstracts were evaluated, and 40 appeared to report the persistence of influenza virus in environmental samples. Evaluation of full texts revealed that 19 of the 40 studies were suitable for review, as they described virus concentration measured at multiple sampling times, with viruses detectable at least twice. Seven studies reported persistence in air (six published before 1970), seven in water (five published after 1990), two in feces, and three on surfaces. All three fomite and five air studies addressed human influenza virus, and all water and feces studies pertained to avian influenza virus. Outcome measurements were transformed to half-lives, and resultant multivariate mixed linear regression models identified influenza virus surviving longer in water than in air. Temperature was a significant predictor of persistence over all matrices. Salinity and pH were significant predictors of persistence in water conditions. An assessment of the methodological quality review of the included studies revealed significant gaps in reporting critical aspects of study design.
Functional and Phylogenetic Analysis of
ureD
in Shiga Toxin-Producing
Escherichia coli
Authors: Steyert Susan R. , Rasko David A. , Kaper James B.
Published: December 2010

Enterohemorrhagic
Escherichia coli
(EHEC) is a food-borne pathogen that can cause severe health complications and utilizes a much lower infectious dose than other
E. coli
pathotypes. Despite having an intact
ure
locus,
ureDABCEFG
, the majority of EHEC strains are phenotypically urease negative under tested conditions. Urease activity potentially assists with survival fitness by enhancing acid tolerance during passage through the stomach or by aiding with colonization in either human or animal reservoirs. Previously, in the EHEC O157:H7 Sakai strain, a point mutation in
ureD
, encoding a urease chaperone protein, was identified, resulting in a substitution of an amber stop codon for glutamine. This single nucleotide polymorphism (SNP) is observed in the majority of EHEC O157:H7 isolates and correlates with a negative urease phenotype
in vitro
. We demonstrate that the lack of urease activity
in vitro
is not solely due to the amber codon in
ureD.
Our analysis has identified two additional SNPs in
ureD
affecting amino acid positions 38 and 205, in both cases determining whether the encoded amino acid is leucine or proline. Phylogenetic analysis based on Ure protein sequences from a variety of urease-encoding bacteria demonstrates that the proline at position 38 is highly conserved among Gram-negative bacteria. Experiments reveal that the L38P substitution enhances urease enzyme activity; however, the L205P substitution does not. Multilocus sequence typing analysis for a variety of Shiga toxin-producing
E. coli
isolates combined with the
ureD
sequence reveals that except for a subset of the O157:H7 strains, neither the
in vitro
urease-positive phenotype nor the
ureD
sequence is phylogenetically restricted.
Three
motAB
Stator Gene Products in
Bdellovibrio bacteriovorus
Contribute to Motility of a Single Flagellum during Predatory and Prey-Independent Growth
Authors: Morehouse Karen A. , Hobley Laura , Capeness Michael , Sockett R. Elizabeth
Published: December 2010

The predatory bacterium
Bdellovibrio bacteriovorus
uses flagellar motility to locate regions rich in Gram-negative prey bacteria, colliding and attaching to prey and then ceasing flagellar motility. Prey are then invaded to form a “bdelloplast” in a type IV pilus-dependent process, and prey contents are digested, allowing
Bdellovibrio
growth and septation. After septation,
Bdellovibrio
flagellar motility resumes inside the prey bdelloplast prior to its lysis and escape of
Bdellovibrio
progeny.
Bdellovibrio
can also grow slowly outside prey as long flagellate host-independent (HI) cells, cultured on peptone-rich media. The
B. bacteriovorus
HD100 genome encodes three pairs of MotAB flagellar motor proteins, each of which could potentially form an inner membrane ion channel, interact with the FliG flagellar rotor ring, and produce flagellar rotation. In 2004, Flannagan and coworkers (R. S. Flannagan, M. A. Valvano, and S. F. Koval, Microbiology 150:649-656, 2004) used antisense RNA and green fluorescent protein (GFP) expression to downregulate a single
Bdellovibrio motA
gene and reported slowed release from the bdelloplast and altered motility of the progeny. Here we inactivated each pair of
motAB
genes and found that each pair contributes to motility, both predatorily, inside the bdelloplast and during HI growth; however, each pair was dispensable, and deletion of no pair abolished motility totally. Driving-ion studies with phenamil, carbonyl cyanide
m
-chlorophenylhydrazone (CCCP), and different pH and sodium conditions indicated that all Mot pairs are proton driven, although the sequence similarities of each Mot pair suggests that some may originate from halophilic species. Thus,
Bdellovibrio
is a “dedicated motorist,” retaining and expressing three pairs of
mot
genes.
The Recombinase IntA Is Required for Excision of
esp
-Containing ICE
Efm1
in
Enterococcus faecium
Authors: Top Janetta , Sinnige Jan C. , Majoor Eline A. M. , Bonten Marc J. M. , Willems Rob J. L. , van Schaik Willem
Published: December 2010

Comparative genome analysis of
Enterococcus faecium
recently revealed that a genomic island containing the
esp
gene, referred to as the
esp
-containing pathogenicity island (
esp
PAI), can be transferred by conjugation and contains a partial Tn
916
-like element and an integrase gene,
intA
. Here, we characterize the role of
intA
in the excision of the
esp
PAI. An
intA
insertion-deletion mutant in
E. faecium
E1162 (E1162Δ
intA
) was constructed and in
trans
complemented with wild-type
intA
(E1162Δ
intA
::pEF30). Circular intermediates (CI) of excised
esp
PAI were determined using inverse PCR analysis on purified chromosomal DNA from strains E1162, E1162Δ
esp
, E1162Δ
intA
, and E1162Δ
intA
::pEF30. In E1162 and E1162Δ
esp
, CI of the
esp
PAI were detected. No CI were detected in E1162Δ
intA
, while in the complemented strain E1162Δ
intA
::pEF30 CI formation was restored, indicating that
intA
is essential for excision and subsequent mobilization of the
esp
-containing genomic island in
E. faecium.
Based on the fact that this island can be mobilized and is self-transmissible, we propose to change the name of the
esp
PAI to ICE
Efm1
.
Mapping Functional Domains of Colicin M
Authors: Helbig Stephanie , Braun Volkmar
Published: December 2010

Colicin M (Cma) lyses
Escherichia coli
cells by inhibiting murein biosynthesis through hydrolysis of the phosphate ester between C
55
-polyisoprenol and
N
-acetylmuramyl (MurNAc)-pentapeptide-GlcNAc in the periplasm. To identify Cma functional domains, we isolated 54 point mutants and small deletion mutants and examined their cytotoxicity levels. Activity and uptake mutants were distinguished by osmotic shock, which transfers Cma into the periplasm independent of the specific FhuA receptor and the Ton system. Deletion of the hydrophobic helix α1, which extends from the compact Cma structure, abolished interference with the antibiotic albomycin, which is transported across the outer membrane by the same system as Cma, thereby identifying α1 as the Cma site that binds to FhuA. Deletion of the C-terminal Lys-Arg strongly reduced Cma translocation across the outer membrane after binding to FhuA. Conversion of Asp226 to Glu, Asn, or Ala inactivated Cma. Asp226 is exposed at the Cma surface and is surrounded by Asp225, Asp229, His235, Tyr228, and Arg236; replacement of each with alanine inactivated Cma. We propose that Asp226 directly participates in phosphate ester hydrolysis and that the surrounding residues contribute to the active site. These residues are strongly conserved in Cma-like proteins of other species. Replacement of other conserved residues with alanine inactivated Cma; these mutations probably altered the Cma structure, as particularly apparent for mutants in the unique open β-barrel of Cma, which were isolated in lower yields. Our results identify regions in Cma responsible for uptake and activity and support the concept of a three-domain arrangement of Cma.
Genome Sequence of
Lactobacillus coryniformis
subsp.
coryniformis
KCTC 3167
Authors: Nam Seong-Hyeuk , Choi Sang-Haeng , Kang Aram , Kim Dong-Wook , Kim Dae-Soo , Kim Ryong Nam , Kim Aeri , Park Hong-Seog
Published: December 2010

Lactobacillus coryniformis
subsp.
coryniformis
is known to be present during the manufacturing process of kimchi, the best-known traditional Korean dish. Here, we present the draft genome sequence of
Lactobacillus coryniformis
subsp.
coryniformis
type strain KCTC 3167 (2,964,752 bp, with a G+C content of 42.8%), which consists of 55 scaffolds.
Complete Genome Sequence of the Bacterium
Methylovorus
sp. Strain MP688, a High-Level Producer of Pyrroloquinolone Quinone
Authors: Xiong Xiang-hua , Zhi Jing-Juan , Yang Lu , Wang Jian-hua , Zhao Yan , Wang Xin , Cui Yan-Jiu , Dong Fang , Li Miao-Xin , Yang Yan-Xin , Wei Na , An Jia-jia , Du Bao-Hua , Liang Long , Zhang Jing-Sheng , Zhou Wei , Cheng Shao-Feng , He Tao , Wang Li , Chen Hui-Peng , Liu Dang-Sheng , Zhang Wei-cai
Published: December 2010

Methylotrophic bacteria are widespread microbes which can use one carbon compound as their only carbon and energy sources. Here we report the finished, annotated genome sequence of the methylotrophic bacterium
Methylovorus
sp. strain MP688, which was isolated from soil for high-level production of pyrroloquinolone quinone (PQQ) in our lab.
The Minor Pilin Subunit Sgp2 Is Necessary for Assembly of the Pilus Encoded by thesrtGCluster ofStreptococcus suis
Authors: Okura Masatoshi , Osaki Makoto , Fittipaldi Nahuel , Gottschalk Marcelo , Sekizaki Tsutomu , Takamatsu Daisuke
Published: December 2010

Gram-positive pili are composed of covalently bound pilin subunits whose assembly is mediated via a pilus-specific sortase(s). Major subunits constitute the pilus backbone and are therefore essential for pilus formation. Minor subunits are also incorporated into the pilus, but they are considered to be dispensable for backbone formation. ThesrtGcluster is one of the putative pilus gene clusters identified in the major swine pathogenStreptococcus suis. It consists of one sortase gene (srtG) and two putative pilin subunit genes (sgp1andsgp2). In this study, by constructing mutants for each of the genes in the cluster and by both immunoblotting and immunogold electron microscopic analysis with antibodies against Sgp1 and Sgp2, we found that thesrtGcluster mediates the expression of pilus-like structures inS. suisstrain 89/1591. In this pilus, Sgp1 forms the backbone, whereas Sgp2 is incorporated as the minor subunit. In accordance with the current model of pilus assembly by Gram-positive organisms, the major subunit Sgp1 was indispensable for backbone formation and the cognate sortase SrtG mediated the polymerization of both subunits. However, unlike other well-characterized Gram-positive bacterial pili, the minor subunit Sgp2 was required for polymerization of the major subunit Sgp1. Because Sgp2 homologues are encoded in several other Gram-positive bacterial pilus gene clusters, in some types of pili, minor pilin subunits may contribute to backbone formation by a novel mechanism.
Correlation between two-dimensional video analysis and subjective assessment in evaluating knee control among elite female team handball players
Authors: Stensrud Silje , Myklebust Grethe , Kristianslund Eirik , Bahr Roald , Krosshaug Tron
Published: December 2010

Poor frontal plane knee control has been shown to be a risk factor for anterior cruciate ligament injuries in ball/team sports. It is necessary to develop simple tests that can identify players with poor knee control and among other factors be able to optimise preventive training programmes. The present study investigated the correlation between a two-dimensional (2D) video analysis and subjective assessment performed by one physiotherapist in evaluating knee control. We also tested the correlation between three simple clinical tests using both methods. A cohort of 186 female elite team handball players completed three tests: single-leg squat (SLS), single-leg vertical drop jump (SLVDJ) and two-leg vertical drop jump (VDJ). Receiver operating characteristic (ROC) analyses showed good to excellent agreement between 2D video analysis and subjective assessment for SLS and VDJ (area under the ROC curve (AUC) 0.83–0.89), but not for SLVDJ (AUC 0.65–0.76). Poor knee control was detected in 25–40% of the players using SLS and VDJ. In contrast, poor knee control was identified in less than 1% of the subjects based on the SLVDJ test. This indicates that SLVDJ may be inadequate for evaluating poor frontal plane knee control. The correlation between the SLS and VDJ tests was found to be low, suggesting that these tests identified different subjects with poor knee control. These results indicate that subjective assessment can be used to screen for poor knee control and that both SLS and VDJ tests should be used in screening athletes for poor knee control.
Increased running speed and previous cramps rather than dehydration or serum sodium changes predict exercise-associated muscle cramping: a prospective cohort study in 210 Ironman triathletes
Authors: Schwellnus Martin P , Drew Nichola , Collins Malcolm
Published: December 2010

Background

Despite the high prevalence of exercise-associated muscle cramping (EAMC) in endurance athletes, the aetiology and risk factors for this condition are not fully understood.

Aim

The aim of this prospective cohort study was to identify risk factors associated with EAMC in endurance triathletes.

Methods

210 triathletes competing in an Ironman triathlon were recruited. Prior to the race, subjects completed a detailed validated questionnaire and blood samples were taken for serum electrolytes. Immediately before the race, pre-race body weight was obtained. Body weight and blood samples for serum electrolyte concentrations were obtained immediately after the race. Clinical data on EAMC experienced during or immediately after the race were also collected.

Results

43 triathletes reported EAMC (cramping group) and were compared with the 166 who did not report EAMC (non-cramping group). There were no significant differences between groups in any pre-race–post-race serum electrolyte concentrations and body weight changes. The development of EAMC was associated with faster predicted race times and faster actual race times, despite similarly matched preparation and performance histories in subjects from both groups. A regression analysis identified faster overall race time (and cycling time) and a history of cramping (in the last 10 races) as the only two independent risk factors for EAMC.

Conclusion

The results from this study add to the evidence that dehydration and altered serum electrolyte balance are not causes for EAMC. Rather, endurance runners competing at a fast pace, which suggests that they exercise at a high intensity, are at risk for EAMC.
Dual role of physical workload and occupational noise in the association of the metabolic syndrome with risk of coronary heart disease: findings from the Helsinki Heart Study
Authors: Koskinen Hanna-Leena , Kauppinen Timo , Tenkanen Leena
Published: December 2010

Objectives

Previous evidence indicates that occupational exposure to physical workload or noise entails development of hypertension and risk of coronary heart disease (CHD). However, vigorous physical activity lessens the risks of the metabolic syndrome (MetS) and CHD. We explored this issue by studying the joint effect of physical workload or noise and MetS on risk of CHD.

Methods

This 18-year follow-up study comprised 1502 middle-aged men employed in industry who participated in the second screening for the Helsinki Heart Study but were not treated with gemfibrozil, the trial drug. The CHD endpoints (ICD-9 codes 410–414 and ICD-10 codes 120–125) were obtained from official Finnish registers. The Finnish job-exposure matrix FINJEM provided information on occupational exposures. The joint effect of baseline MetS levels and both occupational exposures was estimated using Cox's regression models.

Results

Workload and noise increased CHD risk due to increased blood pressure, glucose or body mass index (BMI), separately or combined: the joint effect of workload and MetS defined using these three components yielded an RR of 5.21 (95% CI 2.70 to 10.05). However, when MetS was defined using elevated BMI, high triglycerides and low high-density lipoprotein cholesterol, an RR of 2.19 (95% CI 1.11 to 4.30) among those with MetS only reduced to 1.20 (95% CI 0.61 to 2.35) if concurrently exposed to workload.

Conclusions

Occupational exposure to workload or noise modifies CHD risk differently depending on which definition of MetS is used. In the presence of physical workload or noise, hypertension and blood glucose were the best predictors.