bluuebunny/crossref_metadata_2025 huggingface dataset:
Novel Psychropiezophilic Oceanospirillales Species Profundimonas piezophila gen. nov., sp. nov., Isolated from the Deep-Sea Environment of the Puerto Rico Trench
Authors: Cao Yi , Chastain Roger A. , Eloe Emiley A. , Nogi Yuichi , Kato Chiaki , Bartlett Douglas H.
Published: October 2013

The diversity of deep-sea high-pressure-adapted (piezophilic) microbes in isolated monoculture remains low. In this study, a novel obligately psychropiezophilic bacterium was isolated from seawater collected from the Puerto Rico Trench at a depth of ∼6,000 m. This isolate, designated YC-1, grew best in a nutrient-rich marine medium, with an optimal growth hydrostatic pressure of 50 MPa (range, 20 to 70 MPa) at 8°C. Under these conditions, the maximum growth rate was extremely slow, 0.017 h
−1
, and the maximum yield was 3.51 × 10
7
cells ml
−1
. Cell size and shape changed with pressure, shifting from 4.0 to 5.0 μm in length and 0.5 to 0.8 μm in width at 60 MPa to 0.8- to 1.0-μm diameter coccoid cells under 20 MPa, the minimal pressure required for growth. YC-1 is a Gram-negative, facultatively anaerobic heterotroph. Its predominant cellular fatty acids are the monounsaturated fatty acids (MUFAs) C
16:1
and C
18:1
. Unlike many other psychropiezophiles, YC-1 does not synthesize any polyunsaturated fatty acids (PUFAs). Phylogenetic analysis placed YC-1 within the family of
Oceanospirillaceae
, closely related to the uncultured symbiont of the deep-sea whale bone-eating worms of the genus
Osedax
. In common with some other members of the
Oceanospirillales
, including those enriched during the Deepwater Horizon oil spill, YC-1 is capable of hydrocarbon utilization. On the basis of its characteristics, YC-1 appears to represent both a new genus and a new species, which we name
Profundimonas piezophila
gen. nov., sp. nov.
A New Synthetic Route to
N
-Benzyl Carboxamides through the Reverse Reaction of
N
-Substituted Formamide Deformylase
Authors: Hashimoto Yoshiteru , Sakashita Toshihide , Fukatsu Hiroshi , Sato Hiroyoshi , Kobayashi Michihiko
Published: October 2013

Previously, we isolated a new enzyme,
N
-substituted formamide deformylase, that catalyzes the hydrolysis of
N
-substituted formamide to the corresponding amine and formate (H. Fukatsu, Y. Hashimoto, M. Goda, H. Higashibata, and M. Kobayashi, Proc. Natl. Acad. Sci. U. S. A. 101:13726–13731, 2004,
doi:10.1073/pnas.0405082101
). Here, we discovered that this enzyme catalyzed the reverse reaction, synthesizing
N
-benzylformamide (NBFA) from benzylamine and formate. The reverse reaction proceeded only in the presence of high substrate concentrations. The effects of pH and inhibitors on the reverse reaction were almost the same as those on the forward reaction, suggesting that the forward and reverse reactions are both catalyzed at the same catalytic site. Bisubstrate kinetic analysis using formate and benzylamine and dead-end inhibition studies using a benzylamine analogue, aniline, revealed that the reverse reaction of this enzyme proceeds via an ordered two-substrate, two-product (bi-bi) mechanism in which formate binds first to the enzyme active site, followed by benzylamine binding and the subsequent release of NBFA. To our knowledge, this is the first report of the reverse reaction of an amine-forming deformylase. Surprisingly, analysis of the substrate specificity for acids demonstrated that not only formate, but also acetate and propionate (namely, acids with numbers of carbon atoms ranging from C
1
to C
3
), were active as acid substrates for the reverse reaction. Through this reaction,
N
-substituted carboxamides, such as NBFA,
N
-benzylacetamide, and
N
-benzylpropionamide, were synthesized from benzylamine and the corresponding acid substrates.
Laboratory Evidence of Norwalk Virus Contamination on the Hands of Infected Individuals
Authors: Liu Pengbo , Escudero Blanca , Jaykus Lee-Ann , Montes Julia , Goulter Rebecca M. , Lichtenstein Meredith , Fernandez Marina , Lee Joong-Chul , De Nardo Elizabeth , Kirby Amy , Arbogast James W. , Moe Christine L.
Published: October 2013

Human norovirus (NoV) outbreak investigations suggest that the hands of infected individuals play an important role in NoV transmission. However, there is no experimental evidence documenting the likelihood and degree of NoV contamination on hands. As part of a clinical trial designed to evaluate the efficacy of high-pressure processing for Norwalk virus (NV) inactivation in oysters, 159 hand rinse samples were collected from 6 infected and 6 uninfected subjects. NV was concentrated from the samples by polyethylene glycol precipitation, followed by RNA extraction using an automated guanidinium isothiocyanate-silica method. NV RNA was detected and quantified using multiple NV-specific reverse transcription-quantitative PCR (RT-qPCR) assays. A total of 25.4% (18/71) of the hand rinse samples collected from 6 infected volunteers were presumptively positive for NV, with an average of 3.86 log
10
genomic equivalent copies (GEC) per hand. Dot blot hybridization of PCR products obtained using a different primer set, and DNA sequencing of selected amplicons, provided further confirmation of the presence of NV in the hand rinses. NV contamination was also detected in two hand rinse samples obtained from one uninfected subject. These findings provide definitive evidence of NV contamination on the hands of infected subjects observed under controlled clinical research conditions. Such data support the need for better hand hygiene strategies to prevent NoV transmission.
Aspergillus Collagen-Like Genes (
acl
): Identification, Sequence Polymorphism, and Assessment for PCR-Based Pathogen Detection
Authors: Tuntevski Kiril , Durney Brandon C. , Snyder Anna K. , LaSala P. Rocco , Nayak Ajay P. , Green Brett J. , Beezhold Donald H. , Rio Rita V. M. , Holland Lisa A. , Lukomski Slawomir
Published: October 2013

The genus
Aspergillus
is a burden to public health due to its ubiquitous presence in the environment, its production of allergens, and wide demographic susceptibility among cystic fibrosis, asthmatic, and immunosuppressed patients. Current methods of detection of
Aspergillus
colonization and infection rely on lengthy morphological characterization or nonstandardized serological assays that are restricted to identifying a fungal etiology. Collagen-like genes have been shown to exhibit species-specific conservation across the noncollagenous regions as well as strain-specific polymorphism in the collagen-like regions. Here we assess the conserved region of the
Aspergillus
collagen-like (
acl
) genes and explore the application of PCR amplicon size-based discrimination among the five most common etiologic species of the
Aspergillus
genus, including
Aspergillus fumigatus
,
A. flavus
,
A. nidulans
,
A. niger
, and
A. terreus
. Genetic polymorphism and phylogenetic analysis of the
aclF1
gene were additionally examined among the available strains. Furthermore, the applicability of the PCR-based assay to identification of these five species in cultures derived from sputum and bronchoalveolar fluid from 19 clinical samples was explored. Application of capillary electrophoresis on nanogels was additionally demonstrated to improve the discrimination between
Aspergillus
species. Overall, this study demonstrated that
Aspergillus
acl
genes could be used as PCR targets to discriminate between clinically relevant
Aspergillus
species. Future studies aim to utilize the detection of
Aspergillus
acl
genes in PCR and microfluidic applications to determine the sensitivity and specificity for the identification of
Aspergillus
colonization and invasive aspergillosis in immunocompromised subjects.
SdrA, a New DeoR Family Regulator Involved in Streptomyces avermitilis Morphological Development and Antibiotic Production
Authors: Ulanova Dana , Kitani Shigeru , Fukusaki Eiichiro , Nihira Takuya
Published: October 2013

The SAV3339 (SdrA) protein of
Streptomyces avermitilis
, a member of the DeoR family of regulators, was assessed to determine its
in vivo
function by gene knockdown through the use of
cis
-encoded noncoding RNA and knockout of the
sdrA
gene. These analyses revealed that SdrA represents another class of
Streptomyces
regulator that controls morphological development and antibiotic production.
Dimethyl Adenosine Transferase (KsgA) Deficiency in Salmonella enterica Serovar Enteritidis Confers Susceptibility to High Osmolarity and Virulence Attenuation in Chickens
Authors: Chiok Kim Lam , Addwebi Tarek , Guard Jean , Shah Devendra H.
Published: October 2013

Dimethyl adenosine transferase (KsgA) performs diverse roles in bacteria, including ribosomal maturation and DNA mismatch repair, and synthesis of KsgA is responsive to antibiotics and cold temperature. We previously showed that a
ksgA
mutation in
Salmonella enterica
serovar Enteritidis results in impaired invasiveness in human and avian epithelial cells. In this study, we tested the virulence of a
ksgA
mutant (the
ksgA
::Tn
5
mutant) of
S
. Enteritidis in orally challenged 1-day-old chickens. The
ksgA
::Tn
5
mutant showed significantly reduced intestinal colonization and organ invasiveness in chickens compared to those of the wild-type (WT) parent. Phenotype microarray (PM) was employed to compare the
ksgA
::Tn
5
mutant and its isogenic wild-type strain for 920 phenotypes at 28°C, 37°C, and 42°C. At chicken body temperature (42°C), the
ksgA
::Tn
5
mutant showed significantly reduced respiratory activity with respect to a number of carbon, nitrogen, phosphate, sulfur, and peptide nitrogen nutrients. The greatest differences were observed in the osmolyte panel at concentrations of ≥6% NaCl at 37°C and 42°C. In contrast, no major differences were observed at 28°C. In independent growth assays, the
ksgA
::Tn
5
mutant displayed a severe growth defect in high-osmolarity (6.5% NaCl) conditions in nutrient-rich (LB) and nutrient-limiting (M9 minimum salts) media at 42°C. Moreover, the
ksgA
::Tn
5
mutant showed significantly reduced tolerance to oxidative stress, but its survival within macrophages was not impaired. Unlike
Escherichia coli
, the
ksgA
::Tn
5
mutant did not display a cold-sensitivity phenotype; however, it showed resistance to kasugamycin and increased susceptibility to chloramphenicol. To the best of our knowledge, this is the first report showing the role of
ksgA
in
S
. Enteritidis virulence in chickens, tolerance to high osmolarity, and altered susceptibility to kasugamycin and chloramphenicol.
Resistance of Bacillus subtilis Spore DNA to Lethal Ionizing Radiation Damage Relies Primarily on Spore Core Components and DNA Repair, with Minor Effects of Oxygen Radical Detoxification
Authors: Moeller Ralf , Raguse Marina , Reitz Günther , Okayasu Ryuichi , Li Zuofeng , Klein Stuart , Setlow Peter , Nicholson Wayne L.
Published: October 2013

The roles of various core components, including α/β/γ-type small acid-soluble spore proteins (SASP), dipicolinic acid (DPA), core water content, and DNA repair by apurinic/apyrimidinic (AP) endonucleases or nonhomologous end joining (NHEJ), in
Bacillus subtilis
spore resistance to different types of ionizing radiation including X rays, protons, and high-energy charged iron ions have been studied. Spores deficient in DNA repair by NHEJ or AP endonucleases, the oxidative stress response, or protection by major α/β-type SASP, DPA, and decreased core water content were significantly more sensitive to ionizing radiation than wild-type spores, with highest sensitivity to high-energy-charged iron ions. DNA repair via NHEJ and AP endonucleases appears to be the most important mechanism for spore resistance to ionizing radiation, whereas oxygen radical detoxification via the MrgA-mediated oxidative stress response or KatX catalase activity plays only a very minor role. Synergistic radioprotective effects of α/β-type but not γ-type SASP were also identified, indicating that α/β-type SASP's binding to spore DNA is important in preventing DNA damage due to reactive oxygen species generated by ionizing radiation.
Establishing the Yeast Kluyveromyces lactis as an Expression Host for Production of the Saposin-Like Domain of the Aspartic Protease Cirsin
Authors: Curto Pedro , Lufrano Daniela , Pinto Cátia , Custódio Valéria , Gomes Ana Catarina , Trejo Sebastián A. , Bakás Laura , Vairo-Cavalli Sandra , Faro Carlos , Simões Isaura
Published: October 2013

Typical plant aspartic protease zymogens comprise a characteristic and plant-specific insert (PSI). PSI domains can interact with membranes, and a role as a defensive weapon against pathogens has been proposed. However, the potential of PSIs as antimicrobial agents has not been fully investigated and explored yet due to problems in producing sufficient amounts of these domains in bacteria. Here, we report the development of an expression platform for the production of the PSI domain of cirsin in the generally regarded as safe (GRAS) yeast
Kluyveromyces lactis
. We successfully generated
K. lactis
transformants expressing and secreting significant amounts of correctly processed and glycosylated PSI, as well as its nonglycosylated mutant. A purification protocol with protein yields of ∼4.0 mg/liter was established for both wild-type and nonglycosylated PSIs, which represents the highest reported yield for a nontagged PSI domain. Subsequent bioactivity assays targeting phytopathogenic fungi indicated that the PSI of cirsin is produced in a biologically active form in
K. lactis
and provided clear evidence for its antifungal activity. This yeast expression system thereby emerges as a promising production platform for further exploring the biotechnological potential of these plant saposin-like proteins.
Expression of Functional Plasmodium falciparum Enzymes Using a Wheat Germ Cell-Free System
Authors: Mudeppa Devaraja G. , Rathod Pradipsinh K.
Published: October 2013

One decade after the sequencing of the
Plasmodium falciparum
genome, 95% of malaria proteins in the genome cannot be expressed in traditional cell-based expression systems, and the targets of the best new leads for antimalarial drug discovery are either not known or not available in functional form. For a disease that kills up to 1 million people per year, routine expression of recombinant malaria proteins in functional form is needed both for the discovery of new therapeutics and for identification of targets of new drugs. We tested the general utility of cell-free systems for expressing malaria enzymes. Thirteen test enzyme sequences were reverse amplified from total RNA, cloned into a plant-like expression vector, and subjected to cell-free expression in a wheat germ system. Protein electrophoresis and autoradiography confirmed the synthesis of products of expected molecular masses. In rare problematic cases, truncated products were avoided by using synthetic genes carrying wheat codons. Scaled-up production generated 39 to 354 μg of soluble protein per 10 mg of translation lysate. Compared to rare proteins where cell-based systems do produce functional proteins, the cell-free yields are comparable or better. All 13 test products were enzymatically active, without failure. This general path to produce functional malaria proteins should now allow the community to access new tools, such as biologically active protein arrays, and lead to the discovery of new chemical functions, structures, and inhibitors of previously inaccessible malaria gene products.
Essential Roles for Mycobacterium tuberculosis Rel beyond the Production of (p)ppGpp
Authors: Weiss Leslie A. , Stallings Christina L.
Published: October 2013

In
Mycobacterium tuberculosis
, the stringent response to amino acid starvation is mediated by the
M. tuberculosis
Rel (Rel
Mtb
) enzyme, which transfers a pyrophosphate from ATP to GDP or GTP to synthesize ppGpp and pppGpp, respectively. (p)ppGpp then influences numerous metabolic processes. Rel
Mtb
also encodes a second, distinct catalytic domain that hydrolyzes (p)ppGpp into pyrophosphate and GDP or GTP. Rel
Mtb
is required for chronic
M. tuberculosis
infection in mice; however, it is unknown which catalytic activity of Rel
Mtb
mediates pathogenesis and whether (p)ppGpp itself is necessary. In order to individually investigate the roles of (p)ppGpp synthesis and hydrolysis during
M. tuberculosis
pathogenesis, we generated Rel
Mtb
point mutants that were either synthetase dead (Rel
Mtb
H344Y
) or hydrolase dead (Rel
Mtb
H80A
).
M. tuberculosis
strains expressing the synthetase-dead Rel
Mtb
H344Y
mutant did not persist in mice, demonstrating that the Rel
Mtb
(p)ppGpp synthetase activity is required for maintaining bacterial titers during chronic infection. Deletion of a second predicted (p)ppGpp synthetase had no effect on pathogenesis, demonstrating that Rel
Mtb
was the major contributor to (p)ppGpp production during infection. Interestingly, expression of an allele encoding the hydrolase-dead Rel
Mtb
mutant, Rel
Mtb
H80A
, that is incapable of hydrolyzing (p)ppGpp but still able to synthesize (p)ppGpp decreased the growth rate of
M. tuberculosis
and changed the colony morphology of the bacteria. In addition, Rel
Mtb
H80A
expression during acute or chronic
M. tuberculosis
infection in mice was lethal to the infecting bacteria. These findings highlight a distinct role for Rel
Mtb
-mediated (p)ppGpp hydrolysis that is essential for
M. tuberculosis
pathogenesis.
Identification of a DNA-Damage-Inducible Regulon in Acinetobacter baumannii
Authors: Aranda Jesús , Poza Margarita , Shingu-Vázquez Miguel , Cortés Pilar , Boyce John D. , Adler Ben , Barbé Jordi , Bou Germán
Published: October 2013

The transcriptional response of
Acinetobacter baumannii
, a major cause of nosocomial infections, to the DNA-damaging agent mitomycin C (MMC) was studied using DNA microarray technology. Most of the 39 genes induced by MMC were related to either prophages or encoded proteins involved in DNA repair. Electrophoretic mobility shift assays demonstrated that the product of the
A. baumannii
MMC-inducible
umuD
gene (
umuDAb
) specifically binds to the palindromic sequence TTGAAAATGTAACTTTTTCAA present in its promoter region. Mutations in this palindromic region abolished UmuDAb protein binding. A comparison of the promoter regions of all MMC-induced genes identified four additional transcriptional units with similar palindromic sequences recognized and specifically bound by UmuDAb. Therefore, the UmuDAb regulon consists of at least eight genes encoding seven predicted error-prone DNA polymerase V components and DddR, a protein of unknown function. Expression of these genes was not induced in the MMC-treated
recA
mutant. Furthermore, inactivation of the
umuDAb
gene resulted in the deregulation of all DNA-damage-induced genes containing the described palindromic DNA motif. Together, these findings suggest that UmuDAb is a direct regulator of the DNA damage response in
A. baumannii
.
A Mutation in the N Domain of Escherichia coli Lon Stabilizes Dodecamers and Selectively Alters Degradation of Model Substrates
Authors: Wohlever Matthew L. , Baker Tania A. , Sauer Robert T.
Published: October 2013

Escherichia coli
Lon, an ATP-dependent AAA
+
protease, recognizes and degrades many different substrates, including the RcsA and SulA regulatory proteins. More than a decade ago, the E240K mutation in the N domain of Lon was shown to prevent degradation of RcsA but not SulA
in vivo
. Here, we characterize the biochemical properties of the E240K mutant
in vitro
and present evidence that the effects of this mutation are complex. For example, Lon
E240K
exists almost exclusively as a dodecamer, whereas wild-type Lon equilibrates between hexamers and dodecamers. Moreover, Lon
E240K
displays degradation defects
in vitro
that do not correlate in any simple fashion with degron identity, substrate stability, or dodecamer formation. The Lon sequence segment near residue 240 is known to undergo nucleotide-dependent conformational changes, and our results suggest that this region may be important for coupling substrate binding with allosteric activation of Lon protease and ATPase activity.
Structural and Functional Characterization of an Orphan ATP-Binding Cassette ATPase Involved in Manganese Utilization and Tolerance in Leptospira spp
Authors: Benaroudj Nadia , Saul Frederick , Bellalou Jacques , Miras Isabelle , Weber Patrick , Bondet Vincent , Murray Gerald L. , Adler Ben , Ristow Paula , Louvel Hélène , Haouz Ahmed , Picardeau Mathieu
Published: October 2013

Pathogenic
Leptospira
species are the etiological agents of the widespread zoonotic disease leptospirosis. Most organisms, including
Leptospira
, require divalent cations for proper growth, but because of their high reactivity, these metals are toxic at high concentrations. Therefore, bacteria have acquired strategies to maintain metal homeostasis, such as metal import and efflux. By screening
Leptospira biflexa
transposon mutants for their ability to use Mn
2+
, we have identified a gene encoding a putative orphan ATP-binding cassette (ABC) ATPase of unknown function. Inactivation of this gene in both
L. biflexa
and
L. interrogans
strains led to mutants unable to grow in medium in which iron was replaced by Mn
2+
, suggesting an involvement of this ABC ATPase in divalent cation uptake. A mutation in this ATPase-coding gene increased susceptibility to Mn
2+
toxicity. Recombinant ABC ATPase of the pathogen
L. interrogans
exhibited Mg
2+
-dependent ATPase activity involving a P-loop motif. The structure of this ATPase was solved from a crystal containing two monomers in the asymmetric unit. Each monomer adopted a canonical two-subdomain organization of the ABC ATPase fold with an α/β subdomain containing the Walker motifs and an α subdomain containing the ABC signature motif (LSSGE). The two monomers were arranged in a head-to-tail orientation, forming a V-shaped particle with all the conserved ABC motifs at the dimer interface, similar to functional ABC ATPases. These results provide the first structural and functional characterization of a leptospiral ABC ATPase.
SmbFT, a Putative ABC Transporter Complex, Confers Protection against the Lantibiotic Smb in Streptococci
Authors: Biswas Saswati , Biswas Indranil
Published: October 2013

Streptococcus mutans
, a dental pathogen, secretes different kinds of lantibiotic and nonlantibiotic bacteriocins. For self-protection, a bacteriocin producer strain must possess one or more cognate immunity mechanisms. We report here the identification of one such immunity complex in
S. mutans
strain GS-5 that confers protection against Smb, a two-component lantibiotic. The immunity complex that we identified is an ABC transporter composed of two proteins: SmbF (the ATPase component) and SmbT (the permease component). Both of the protein-encoding genes are located within the
smb
locus. We show that GS-5 becomes sensitized to Smb upon deletion of
smbT
, which makes the ABC transporter nonfunctional. To establish the role SmbFT in providing immunity, we heterologously expressed this ABC transporter complex in four different sensitive streptococcal species and demonstrated that it can confer resistance against Smb. To explore the specificity of SmbFT in conferring resistance, we tested mutacin IV (a nonlantibiotic), nisin (a single peptide lantibiotics), and three peptide antibiotics (bacitracin, polymyxin B, and vancomycin). We found that SmbFT does not recognize these structurally different peptides. We then tested whether SmbFT can confer protection against haloduracin, another two-component lantibiotic that is structurally similar to Smb; SmbFT indeed conferred protection against haloduracin. SmbFT can also confer protection against an uncharacterized but structurally similar lantibiotic produced by
Streptococcus gallolyticus
. Our data suggest that SmbFT truly displays immunity function and confer protection against Smb and structurally similar lantibiotics.
Variables associated with return to sport following anterior cruciate ligament reconstruction: a systematic review
Authors: Czuppon Sylvia , Racette Brad A , Klein Sandra E , Harris-Hayes Marcie
Published: October 2013

Background

As one of the purposes of anterior cruciate ligament reconstruction (ACLR) is to return athletes to their preinjury activity level, it is critical to understand variables influencing return to sport. Associations between return to sport and variables representing knee impairment, function and psychological status have not been well studied in athletes following ACLR.

Purpose

The purpose of this review was to summarise the literature reporting on variables proposed to be associated with return to sport following ACLR.

Study design

Systematic review.

Methods

Medline, EMBASE, CINAHL and Cochrane databases were searched for articles published before November 2012. Articles included in this review met these criteria: (1) included patients with primary ACLR, (2) reported at least one knee impairment, function or psychological measure, (3) reported a return to sport measure and (4) analysed the relationship between the measure and return to sport.

Results

Weak evidence existed in 16 articles suggesting variables associated with return to sport included higher quadriceps strength, less effusion, less pain, greater tibial rotation, higher Marx Activity score, higher athletic confidence, higher preoperative knee self-efficacy, lower kinesiophobia and higher preoperative self-motivation.

Conclusions

Weak evidence supports an association between knee impairment, functional and psychological variables and return to sport. Current return to sport guidelines should be updated to reflect all variables associated with return to sport. Utilising evidence-based return to sport guidelines following ACLR may ensure that athletes are physically and psychologically capable of sports participation, which may reduce reinjury rates and the need for subsequent surgery.
The financial cost of hamstring strain injuries in the Australian Football League
Authors: Hickey Jack , Shield Anthony J , Williams Morgan D , Opar David A
Published: October 2013

Background

Hamstring strain injuries (HSIs) have remained the most prevalent injury in the Australian Football League (AFL) over the past 21 regular seasons. The effect of HSIs in sports is often expressed as regular season games missed due to injury. However, the financial cost of athletes missing games due to injury has not been investigated. The aim of this report is to estimate the financial cost of games missed due to HSIs in the AFL.

Method

Data were collected using publicly available information from the AFL's injury report and the official AFL annual report for the past 10 competitive AFL seasons. Average athlete salary and injury epidemiology data were used to determine the average yearly financial cost of HSIs for AFL clubs and the average financial cost of a single HSI over this time period.

Results

Across the observed period, average yearly financial cost of HSIs per club increased by 71% compared with a 43% increase in average yearly athlete salary. Over the same time period the average financial cost of a single HSI increased by 56% from $A25 603 in 2003 to $A40 021 in 2012, despite little change in the HSI rates during the period.

Conclusions

The observed increased financial cost of HSIs was ultimately explained by the failure of teams to decrease HSI rates, but coupled with increases in athlete salaries over the past 10 season. The information presented in this report highlights the financial cost of HSIs and other sporting injuries, raising greater awareness and the need for further funding for research into injury prevention strategies to maximise economical return for investment in athletes.
The Application of Intermediate Temperature Fuel Cell for the Auxiliary Power Unit of the Air Conditioning System in an Electric Vehicle
Authors: Pethaiah Sethu Sundar , Subiantoro Alison , Stimming Ulrich
Published: October 2013

One of the key research challenges in E-car technology is the air conditioning system, which draws up tremendous electrical energy from the battery system when in operation. A typical electrical vehicle would have a travel range of 150km, but this range may drop to 80km when the air-conditioner is switched on. To address this issue, on one hand, the proposed work discusses the possible ways to reduce the power consumption of air conditioning system in Electric Vehicles and the applications of intermediate temperature PEMFC as an APU. A typical intermediate temperature PEMFC operates at 150-250°C using ethanol as the feedstock. Ethanol is a promising fuel for future transport applications because of its low toxicity and high energy content [6.3 kWh/L, which is higher than H2 (@1000 psi, 25oC, 0.18 kWh/L) and methanol (4.8kWh/L)].
Behavior of a Single Direct Methanol Fuel Cell in Stacks at Air Maldistribution Conditions
Authors: Yang Xiao-Guang , Ye Qiang , Cheng Ping
Published: October 2013

The air maldistribution through different individual cells is frequently encountered in direct methanol fuel cell (DMFC) stacks since most stacks utilize parallel flow configuration. This air maldistribution has been recognized as one of the most important causes for the performance loss and even durability loss of DMFC stacks. To better understand the impact of air maldistribution, the behavior of a single DMFC at various air flow rates (AFRs) is investigated both experimentally and numerically. Special attention is paid to the variation of cell voltage with AFR. It is found that the cell voltage drops significantly when the AFR decreases below a critical value. With the further drop of AFR, the cell voltage descends more sharply and even becomes negative at ultralow AFRs. The reasons for this unique behavior of cell voltage with AFR are explored in detail within this work.
Effect of Electrode Pretreatment on the Cyclic Voltammetry of VO2+/ VO2
+ at a Glassy Carbon Electrode
Authors: Bourke Andrea , Lynch Robert P. , Buckley D. Noel
Published: October 2013

It is shown by cyclic voltammetry and current measurements at constant potential that, when glassy carbon electrodes are pretreated by polarization at potentials more positive than 0.5 V (sat. Hg/Hg2SO4), they are subsequently less active for oxidation of VO2+. The activity of the electrode is recovered by polarization at negative potentials. At any given potential a characteristic amount of electrode deactivation occurs in a relatively short time (~60 s) and further pretreatment at that potential does not further deactivate the electrode. It is suggested that the level of deactivation is determined by processes that are primarily dependent on potential rather than on kinetics and involve the formation of oxygen species that interact with active sites on the carbon surface.
High Performance LiNi0.5Mn1.5O4 Spinel Li-Ion Battery Cathode Development
Authors: Xing Weibing , Buettner-Garrett Josh , Krysiak Mike , Kelly Joseph
Published: October 2013

Due to its high theoretical capacity (147 mAh/g), high operating voltage (~4.7V vs. Li/Li+ of Ni2+/Ni4+ couple), and inherent cycle stability (Mn4+ state, no Jahn-Teller distortion of Mn3+), LiNi0.5Mn1.5O4 (LNMO) spinel is a promising cathode active material for high energy density lithium ion batteries [1, 2, 3] that can potentially be used in a wide range of applications including consumer electronics, electric vehicles, military and space. However, some performance issues of LNMO need to be addressed before its practical commercialization. These include extended cycle life, high rate capability, and cycle stability at elevated temperatures (e.g. 55°C).Here we report efforts at ADA Technologies, Inc. to optimize LiNi0.5Mn1.5O4 (LNMO) spinel performance via material and operational approaches. In addition to improved stability with the electrolyte [4], oxide coatings applied via atomic layer deposition (ALD) technique are used to address cycle stability of LNMO at high temperatures.
Thermodynamics and Kinetics of Solid Oxide Fuel Cells with Molten Tin Anodes
Authors: Colet Lagrille Melanie , Kelsall Geoff H.
Published: October 2013

Carbon-air fuel cells are a prospective technology for efficient and environmentally-benign conversion of the chemical energy of solid carbonaceous fuels directly to electrical energy. Since Ni-YSZ anodes used in conventional (H2/O2) solid oxide fuel cells (SOFCs) are irreversibly affected by sulphur poisoning and deposition of carbon, some molten metal anodes may offer better alternatives, with high electrical conductivities and stabilities when operating on (impure) carbonaceous fuels. Tin fulfils these requirements and is non-toxic, has a low melting point and vapour pressure, forms oxide(s) that can be reduced by CO (or H2), and could be purified of ash, which it would not wet, and of dissolved metals by electrochemical processes. The thermodynamics and kinetics of a solid oxide fuel cell with molten tin anode (Sn(l)-SOFC) have been studied by cyclic voltammetry and electrochemical impedance spectroscopy to determine the reactions occurring in the fuel cell when operating at 900 °C in battery mode and the kinetic parameters of the anode oxidation reaction. Results suggested that the oxidation | reduction processes taking place at the Sn(l)-SOFC are controlled by slow diffusion of oxygen species ([O]Sn) at the molten tin anode and the resulting formation/accumulation of tin dioxide (SnO2) at the anode | electrolyte interface.
Preparation and Characterization of La0.9Sr0.1Ga0.8Mg0.2O3
-δ Thin Film Electrolyte Prepared by Spray Pyrolysis Deposition for Low-Temperature Operating µ-SOFC
Authors: Tomomichi Keita , Sasaki Kazuya , Endo Yoichi , Kikai Kohei , Terai Takayuki
Published: October 2013

Spray pyrolysis deposition (SPD) is a potential method for mass production of ceramics thin film with low fabrication cost. We investigated on the SPD method, and demonstrated that it is a noble method to prepare the electrolyte thin film for solid oxide fuel cells. Controlling the substrate temperature low to about 150 oC during the SPD process is a key factor, in order to make thin film dense and continued. Repeating SPD and post annealing is effective to fill up micro cracks which happened due to the difference of coefficients of thermal expansion between the substrate and the film.
Empirical Correlations to Predict In-situ Durability of Polymer Electrolyte Membranes in Fuel Cells
Authors: Yadav R. , DiLeo G. , Dale N. , Adjemian K.
Published: October 2013

Chemical and mechanical durability of polymer electrolyte membrane (PEM) must be understood and improved for commercial success of PEMFCs vehicles. Chemical degradation of PEMs occurs under OCV conditions due to high gas crossover, high concentration of chemically weak groups, and low EW. These properties measured ex-situ have been related to develop a chemical stability factor (CSF) to predict in-situ chemical durability of PEMs. Mechanical degradation of PEMs occurs under dry-wet cycling due to low tensile strain, high swelling, low EW and high gas crossover. A mechanical stability factor (MSF) has been derived from ex-situ measured % Strain at Break, % Swelling, oxygen crossover rate, and EW to predict in-situ mechanical durability of PEMs. The relative trend predicted by CSF and MSF follows the relative trend in in-situ chemical and mechanical durability. These factors can be used to screen and select the most durable PEM for in-situ testing in order to save lengthy in-situ testing time, developmental cost and resources.
Energy Recovery in Parallel Capacitive Deionization Operations
Authors: Landon James , Gao Xin , Neathery James K. , Liu Kunlei
Published: October 2013

The integration of capacitive deionization cells is shown to deionize a secondary salt stream using solely the energy stored in a charged CDI cell. A primary cell was charged at 1.2 and 1.5 V resulting in electrosorption capacities of 3.3 and 3.7 mg NaCl (g carbon)-1, respectively. The cell charged at 1.2 and 1.5 V was used to directly deionize a parallel secondary stream resulting in electrosorption capacities of 0.5 and 1.0 mg NaCl (g carbon)-1, respectively. Integration of CDI cells can drastically lower the energy cost for the production of fresh water.
Perovskite Ba0.5Sr0.5Co0.8Fe0.2O3–δ as a Bifunctional Air Electrode for Zinc-Air Batteries
Authors: Yuan Xiao-Zi , Qu Wei , Fahlman Jason , Ivey Douglas G. , Zhang Xinge
Published: October 2013

Perovskite Ba0.5Sr0.5Co0.8Fe0.2O3–δ (BSCF) powder was prepared through a sol-gel method and characterized by X-ray diffraction (XRD), Brunauer–Emmett–Teller (BET) analysis and transmission electron microscopy (TEM). The electrocatalytic properties of BSCF/C composites towards the oxygen reduction reaction (ORR) and the oxygen evolution reaction (OER) were studied using the rotating disk electrode (RDE) technique in 6 M KOH. The results showed that the synthesized perovskite BSCF exhibits activities for both ORR and OER, especially for OER. Cell tests demonstrated that the synthesized material reaches an ORR activity of 83 mAcm-2 at a potential of 0.9 V and an OER activity of 117 mAcm-2 at a potential of 2.2 V at about the 100th cycle. With increasing cycle numbers, both OER and ORR activities had noticeable degradation.
A novel approach to assess hemorrhagic shock severity using the arterially determined left ventricular isovolumic contraction period
Authors: van Houwelingen Marc J. , Merkus Daphne , Hofland Jan , Bakker Jan , Tenbrinck Robert , te Lintel Hekkert Maaike , van Dijk Geert , Hoeks Arnold P. G. , Duncker Dirk J.
Published: October 2013

Recently, the ventilatory variation in pre-ejection period (ΔPEP) was found to be useful in the prediction of fluid-responsiveness of patients in shock. In the present study we investigated the behavior of the ventilation-induced variations in the systolic timing intervals in response to a graded hemorrhage protocol. The timing intervals studied included the ventilatory variation in ventricular electromechanical delay (ΔEMD), isovolumic contraction period (determined from the arterial pressure waveform, ΔAIC), pulse travel time (ΔPTT), and ΔPEP. ΔAIC and ΔPEP were evaluated in the aorta and carotid artery (annotated by subscripts Ao and CA) and were compared with the responses of pulse pressure variation (ΔPPAo) and stroke volume variation (ΔSV). The graded hemorrhage protocol, followed by resuscitation using norepinephrine and autologous blood transfusion, was performed in eight anesthetized Yorkshire X Landrace swine. ΔAICAo, ΔAICCA, ΔPEPAo, ΔPEPCA, ΔPPAo, ΔPPCA, and ΔSV showed significant increases during the graded hemorrhage and significant decreases during the subsequent resuscitation. ΔAICAo, ΔAICCA, ΔPEPAo, and ΔPEPCA all correlated well with ΔPPAo and ΔSV (all r ≥ 0.8, all P < 0.001). ΔEMD and ΔPTT did not significantly change throughout the protocol. In contrast with ΔPEPAo, which was significantly higher than ΔPEPCA ( P < 0.01), ΔAICAo was not different from ΔAICCA. In conclusion, ventilation-induced preload variation principally affects the arterially determined isovolumic contraction period (AIC). Moreover, ΔAIC can be determined solely from the arterial pressure waveform, whereas ΔPEP also requires ECG measurement. Importantly, ΔAIC determined from either the carotid or aortic pressure waveform are interchangeable, suggesting that, in contrast with ΔPEP, ΔAIC may be site independent.
Pulmonary artery endothelial cell dysfunction and decreased populations of highly proliferative endothelial cells in experimental congenital diaphragmatic hernia
Authors: Acker Shannon N. , Seedorf Gregory J. , Abman Steven H. , Nozik-Grayck Eva , Partrick David A. , Gien Jason
Published: October 2013

Decreased lung vascular growth and pulmonary hypertension contribute to poor outcomes in congenital diaphragmatic hernia (CDH). Mechanisms that impair angiogenesis in CDH are poorly understood. We hypothesize that decreased vessel growth in CDH is caused by pulmonary artery endothelial cell (PAEC) dysfunction with loss of a highly proliferative population of PAECs (HP-PAEC). PAECs were harvested from near-term fetal sheep that underwent surgical disruption of the diaphragm at 60–70 days gestational age. Highly proliferative potential was measured via single cell assay. PAEC function was assessed by assays of growth and tube formation and response to known proangiogenic stimuli, vascular endothelial growth factor (VEGF), and nitric oxide (NO). Western blot analysis was used to measure content of angiogenic proteins, and superoxide production was assessed. By single cell assay, the proportion of HP-PAEC with growth of >1,000 cells was markedly reduced in the CDH PAEC, from 29% (controls) to 1% (CDH) ( P < 0.0001). Compared with controls, CDH PAEC growth and tube formation were decreased by 31% ( P = 0.012) and 54% ( P < 0.001), respectively. VEGF and NO treatments increased CDH PAEC growth and tube formation. VEGF and VEGF-R2 proteins were increased in CDH PAEC; however, eNOS and extracellular superoxide dismutase proteins were decreased by 29 and 88%, respectively. We conclude that surgically induced CDH in fetal sheep causes endothelial dysfunction and marked reduction of the HP-PAEC population. We speculate that this CDH PAEC phenotype contributes to impaired vascular growth in CDH.
Soluble Carcinoembryonic Antigen Activates Endothelial Cells and Tumor Angiogenesis
Authors: Bramswig Kira H. , Poettler Marina , Unseld Matthias , Wrba Friedrich , Uhrin Pavel , Zimmermann Wolfgang , Zielinski Christoph C. , Prager Gerald W.
Published: October 2013

Carcinoembryonic antigen (CEA, CD66e, CEACAM-5) is a cell-surface–bound glycoprotein overexpressed and released by many solid tumors that has an autocrine function in cancer cell survival and differentiation. Soluble CEA released by tumors is present in the circulation of patients with cancer, where it is used as a marker for cancer progression, but whether this form of CEA exerts any effects in the tumor microenvironment is unknown. Here, we present evidence that soluble CEA is sufficient to induce proangiogenic endothelial cell behaviors, including adhesion, spreading, proliferation, and migration in vitro and tumor microvascularization in vivo. CEA-induced activation of endothelial cells was dependent on integrin β-3 signals that activate the focal-adhesion kinase and c-Src kinase and their downstream MAP–ERK kinase/extracellular signal regulated kinase and phosphoinositide 3-kinase/Akt effector pathways. Notably, while interference with VEGF signaling had no effect on CEA-induced endothelial cell activation, downregulation with the CEA receptor in endothelial cells attenuated CEA-induced signaling and tumor angiogenesis. Corroborating these results clinically, we found that tumor microvascularization was higher in patients with colorectal cancer exhibiting higher serum levels of soluble CEA. Together, our results elucidate a novel function for soluble CEA in tumor angiogenesis. Cancer Res; 73(22); 6584–96. ©2013 AACR.
Maintenance of Androgen Receptor Inactivation by S-Nitrosylation
Authors: Qin Yu , Dey Anindya , Purayil Hamsa Thayele , Daaka Yehia
Published: October 2013

Antiandrogens target ligand-binding domain of androgen receptor (AR) and are used as first-line therapeutics to treat patients diagnosed with locally advanced and metastatic prostate cancer. Although initially beneficial as judged with actual tumor mass shrinkage, this therapy invariably fails and the cancer reappears as castration-resistant disease. Here, we report that increased intracellular nitric oxide (NO) levels lead to growth inhibition of both androgen-dependent and castration-resistant prostate tumors through a mechanism that involves AR function inactivation by S-nitrosylation of a single C601 residue present in the DNA-binding domain. AR S-nitrosylation does not impact its subcellular distribution but attenuates its ability to bind AR-responsive elements in promoter region of target genes. Mechanistically, AR is transnitrosylated by its partner HSP90 protein. Ubiquitous small-molecule NO donors promote the AR S-nitrosylation and inhibit growth of castration-resistant prostate tumors. These findings reveal a new mechanism of regulating AR function and suggest that sequential targeting of distinct domains of AR may extend therapeutic efficacy for patients with advanced prostate cancer. Cancer Res; 73(22); 6690–9. ©2013 AACR.
Carbon Monoxide Expedites Metabolic Exhaustion to Inhibit Tumor Growth
Authors: Wegiel Barbara , Gallo David , Csizmadia Eva , Harris Clair , Belcher John , Vercellotti Gregory M. , Penacho Nuno , Seth Pankaj , Sukhatme Vikas , Ahmed Asif , Pandolfi Pier Paolo , Helczynski Leszek , Bjartell Anders , Persson Jenny Liao , Otterbein Leo E.
Published: October 2013

One classical feature of cancer cells is their metabolic acquisition of a highly glycolytic phenotype. Carbon monoxide (CO), one of the products of the cytoprotective molecule heme oxygenase-1 (HO-1) in cancer cells, has been implicated in carcinogenesis and therapeutic resistance. However, the functional contributions of CO and HO-1 to these processes are poorly defined. In human prostate cancers, we found that HO-1 was nuclear localized in malignant cells, with low enzymatic activity in moderately differentiated tumors correlating with relatively worse clinical outcomes. Exposure to CO sensitized prostate cancer cells but not normal cells to chemotherapy, with growth arrest and apoptosis induced in vivo in part through mitotic catastrophe. CO targeted mitochondria activity in cancer cells as evidenced by higher oxygen consumption, free radical generation, and mitochondrial collapse. Collectively, our findings indicated that CO transiently induces an anti-Warburg effect by rapidly fueling cancer cell bioenergetics, ultimately resulting in metabolic exhaustion. Cancer Res; 73(23); 7009–21. ©2013 AACR.
The HSP90 Inhibitor Ganetespib Synergizes with the MET Kinase Inhibitor Crizotinib in both Crizotinib-Sensitive and -Resistant MET-Driven Tumor Models
Authors: Miyajima Naoto , Tsutsumi Shinji , Sourbier Carole , Beebe Kristin , Mollapour Mehdi , Rivas Candy , Yoshida Soichiro , Trepel Jane B. , Huang Ying , Tatokoro Manabu , Shinohara Nobuo , Nonomura Katsuya , Neckers Len
Published: October 2013

The proto-oncogene MET is aberrantly activated via overexpression or mutation in numerous cancers, making it a prime anticancer molecular target. However, the clinical success of MET-directed tyrosine kinase inhibitors (TKI) has been limited due, in part, to mutations in the MET kinase domain that confer therapeutic resistance. Circumventing this problem remains a key challenge to improving durable responses in patients receiving MET-targeted therapy. MET is an HSP90-dependent kinase, and in this report we show that HSP90 preferentially interacts with and stabilizes activated MET, regardless of whether the activation is ligand-dependent or is a consequence of kinase domain mutation. In contrast, many MET-TKI show a preference for the inactive form of the kinase, and activating mutations in MET can confer resistance. Combining the HSP90 inhibitor ganetespib with the MET-TKI crizotinib achieves synergistic inhibition of MET, its downstream signaling pathways, and tumor growth in both TKI-sensitive and -resistant MET-driven tumor models. These data suggest that inclusion of an HSP90 inhibitor can partially restore TKI sensitivity to previously resistant MET mutants, and they provide the foundation for clinical evaluation of this therapeutic combination in patients with MET-driven cancers. Cancer Res; 73(23); 7022–33. ©2013 AACR.