bluuebunny/crossref_metadata_2025_split huggingface dataset:
Vaginal Histological Changes after Using Intravaginal Sponges for Oestrous Synchronization in Anoestrous Ewes
Authors: Manes J , Campero C , Hozbor F , Alberio R , Ungerfeld R 
Published: January 2015

ContentsTo characterize the histological and cytological vaginal changes generated by the use of intravaginal sponge (IS) applied in oestrous synchronization treatments in ewes during mid‐non‐breeding season. Thirty‐five multiparous ewes were allocated to three experimental groups according to the moment in which the samples were taken: (i) ewes treated with IS containing 60 mg of medroxyprogesterone acetate for 14 days, sampled the day of IS removal (group ISR; n = 10), (ii) or after sponge removal at time of oestrus or 72 h after removal (group AR; n = 14) and (iii) ewes without sponge treatment that were sampled at the day of IS removal of the other groups (group CG; n = 11). Vaginal biopsies and cytological samples were taken from the anterior vaginal fornix area. The vagina of the CG group had a stratified squamous epithelium with a moderate degree of cellular infiltration with lymphocytes and plasma cells in the lamina propia. Treated ewes (ISR and AR) had epithelial hyperplasia and hypertrophy. ISR ewes had haemorrhage and perivascular infiltrate and an increased number of epithelial cells, neutrophils, macrophages and erythrocytes at IS removal. The use of IS generated histological and cytological alterations in the vaginal wall when used for oestrous synchronization in anoestrous ewes.
Nitric oxide selectively suppresses IH currents mediated by HCN1‐containing channels
Authors: Kopp‐Scheinpflug Cornelia , Pigott Beatrice M. , Forsythe Ian D. 
Published: January 2015

Key points
The superior olivary complex (SOC) exhibits a spectrum of HCN1 and HCN2 subunit expression, which generate IH currents with fast and slow kinetics, respectively.
Neuronal nitric oxide synthase (nNOS) was broadly distributed across the SOC.
NO hyperpolarizes the half‐activation voltage of HCN1‐mediated currents and caused a slowing of the IH current kinetics in the respective nuclei (medial and lateral superior olives and superior paraolivary nucleus). This signalling was independent of cGMP.
NO also caused a depolarizing shift in the half‐activation voltage of HCN2‐mediated IH currents, increasing activation at resting potentials; this was cGMP‐dependent.
Thus, NO signalling suppressed fast HCN1‐mediated currents and potentiated slow HCN2‐mediated currents, modulating the overall kinetics and magnitude of the endogenous IH.
AbstractHyperpolarization‐activated non‐specific cation‐permeable channels (HCN) mediate IH currents, which are modulated by cGMP and cAMP and by nitric oxide (NO) signalling. Channel properties depend upon subunit composition (HCN1–4 and accessory subunits) as demonstrated in expression systems, but physiological relevance requires investigation in native neurons with intact intracellular signalling. Here we use the superior olivary complex (SOC), which exhibits a distinctive pattern of HCN1 and HCN2 expression, to investigate NO modulation of the respective IH currents, and compare properties in wild‐type and HCN1 knockout mice. The medial nucleus of the trapezoid body (MNTB) expresses HCN2 subunits exclusively, and sends inhibitory projections to the medial and lateral superior olives (MSO, LSO) and the superior paraolivary nucleus (SPN). In contrast to the MNTB, these target nuclei possess an IH with fast kinetics, and they express HCN1 subunits. NO is generated in the SOC following synaptic activity and here we show that NO selectively suppresses HCN1, while enhancing IH mediated by HCN2 subunits. NO hyperpolarizes the half‐activation of HCN1‐mediated currents and slows the kinetics of native IH currents in the MSO, LSO and SPN. This modulation was independent of cGMP and absent in transgenic mice lacking HCN1. Independently, NO signalling depolarizes the half‐activation of HCN2‐mediated IH currents in a cGMP‐dependent manner. Thus, NO selectively suppresses fast HCN1‐mediated IH and facilitates a slow HCN2‐mediated IH, so generating a spectrum of modulation, dependent on the local expression of HCN1 and/or HCN2.
Molecular underpinnings of ventral surface chemoreceptor function: focus on KCNQ channels
Authors: Mulkey Daniel K. , Hawkins Virginia E. , Hawryluk Joanna M. , Takakura Ana C. , Moreira Thiago S. , Tzingounis Anastasios V. 
Published: January 2015

AbstractCentral chemoreception is the mechanism by which CO2/H+‐sensitive neurons (i.e. chemoreceptors) regulate breathing in response to changes in tissue CO2/H+. Neurons in the retrotrapezoid nucleus (RTN) directly regulate breathing in response to changes in tissue CO2/H+ and function as a key locus of respiratory control by integrating information from several respiratory centres, including the medullary raphe. Therefore, chemosensitive RTN neurons appear to be critically important for maintaining breathing, thus understanding molecular mechanisms that regulate RTN chemoreceptor function may identify therapeutic targets for the treatment of respiratory control disorders. We have recently shown that KCNQ (Kv7) channels in the RTN are essential determinants of spontaneous activity ex vivo, and downstream effectors for serotonergic modulation of breathing. Considering that loss of function mutations in KCNQ channels can cause certain types of epilepsy including those associated with sudden unexplained death in epilepsy (SUDEP), we propose that dysfunctions of KCNQ channels may be one cause for epilepsy and respiratory problems associated with SUDEP. In this review, we will summarize the role of KCNQ channels in the regulation of RTN chemoreceptor function, and suggest that these channels represent useful therapeutic targets for the treatment of respiratory control disorders.
KCNQ1 channel modulation by KCNE proteins via the voltage‐sensing domain
Authors: Nakajo Koichi , Kubo Yoshihiro 
Published: January 2015

AbstractThe gating of the KCNQ1 potassium channel is drastically regulated by auxiliary subunit KCNE proteins. KCNE1, for example, slows the activation kinetics of KCNQ1 by two orders of magnitude. Like other voltage‐gated ion channels, the opening of KCNQ1 is regulated by the voltage‐sensing domain (VSD; S1–S4 segments). Although it has been known that KCNE proteins interact with KCNQ1 via the pore domain, some recent reports suggest that the VSD movement may be altered by KCNE. The altered VSD movement of KCNQ1 by KCNE proteins has been examined by site‐directed mutagenesis, the scanning cysteine accessibility method (SCAM), voltage clamp fluorometry (VCF) and gating charge measurements. These accumulated data support the idea that KCNE proteins interact with the VSDs of KCNQ1 and modulate the gating of the KCNQ1 channel. In this review, we will summarize recent findings and current views of the KCNQ1 modulation by KCNE via the VSD. In this context, we discuss our recent findings that KCNE1 may alter physical interactions between the S4 segment (VSD) and the S5 segment (pore domain) of KCNQ1. Based on these findings from ourselves and others, we propose a hypothetical mechanism for how KCNE1 binding alters the VSD movement and the gating of the channel.
The Effectiveness of Foot Orthotics in Improving Postural Control in Individuals With Chronic Ankle Instability: A Critically Appraised Topic
Authors: Gabriner Michael L. , Braun Brittany A. , Houston Megan N. , Hoch Matthew C. 
Published: January 2015

Clinical Scenario:Chronic ankle instability (CAI) is a condition commonly experienced by physically active individuals. It has been suggested that foot orthotics may increase a CAI patient’s postural control.Clinical Question:For patients with CAI, is there evidence to suggest that an orthotic intervention will help improve postural control?Summary of Key Findings:The literature was searched for studies of level 2 evidence or higher that investigated the effects of foot orthotics on postural control in patients with CAI. The search of the literature produced 5 possible studies for inclusion; 2 studies met the inclusion criteria and were included. One randomized controlled trial and 1 outcomes study were included. Foot orthotics appear to be effective at improving postural control in patients with CAI.Clinical Bottom Line:There is moderate evidence to support the use of foot orthotics in the treatment of CAI to help improve postural control.Strength of Recommendation:There is grade B evidence that foot orthotics help improve postural control in people with CAI. The Centre of Evidence Based Medicine recommends a grade of B for level 2 evidence with consistent findings.
Novel Phage Lysin Capable of Killing the Multidrug-Resistant Gram-Negative Bacterium Acinetobacter baumannii in a Mouse Bacteremia Model
Authors: Lood Rolf , Winer Benjamin Y. , Pelzek Adam J. , Diez-Martinez Roberto , Thandar Mya , Euler Chad W. , Schuch Raymond , Fischetti Vincent A. 
Published: January 2015

ABSTRACT

Acinetobacter baumannii
, a Gram-negative multidrug-resistant (MDR) bacterium, is now recognized as one of the more common nosocomial pathogens. Because most clinical isolates are found to be multidrug resistant, alternative therapies need to be developed to control this pathogen. We constructed a bacteriophage genomic library based on prophages induced from 13
A. baumannii
strains and screened it for genes encoding bacteriolytic activity. Using this approach, we identified 21 distinct lysins with different activities and sequence diversity that were capable of killing
A. baumannii
. The lysin (PlyF307) displaying the greatest activity was further characterized and was shown to efficiently kill (>5-log-unit decrease) all tested
A. baumannii
clinical isolates. Treatment with PlyF307 was able to significantly reduce planktonic and biofilm
A. baumannii
both
in vitro
and
in vivo
. Finally, PlyF307 rescued mice from lethal
A. baumannii
bacteremia and as such represents the first highly active therapeutic lysin specific for Gram-negative organisms in an array of native lysins found in
Acinetobacter
phage.
Combinational Deletion of Three Membrane Protein-Encoding Genes Highly Attenuates Yersinia pestis while Retaining Immunogenicity in a Mouse Model of Pneumonic Plague
Authors: Tiner Bethany L. , Sha Jian , Kirtley Michelle L. , Erova Tatiana E. , Popov Vsevolod L. , Baze Wallace B. , van Lier Christina J. , Ponnusamy Duraisamy , Andersson Jourdan A. , Motin Vladimir L. , Chauhan Sadhana , Chopra Ashok K. 
Published: January 2015

ABSTRACT

Previously, we showed that deletion of genes encoding Braun lipoprotein (Lpp) and MsbB attenuated
Yersinia pestis
CO92 in mouse and rat models of bubonic and pneumonic plague. While Lpp activates Toll-like receptor 2, the MsbB acyltransferase modifies lipopolysaccharide. Here, we deleted the
ail
gene (encoding the
a
ttachment-
i
nvasion
l
ocus) from wild-type (WT) strain CO92 or its
lpp
single and Δ
lpp
Δ
msbB
double mutants. While the Δ
ail
single mutant was minimally attenuated compared to the WT bacterium in a mouse model of pneumonic plague, the Δ
lpp
Δ
ail
double mutant and the Δ
lpp
Δ
msbB
Δ
ail
triple mutant were increasingly attenuated, with the latter being unable to kill mice at a 50% lethal dose (LD
50
) equivalent to 6,800 LD
50
s of WT CO92. The mutant-infected animals developed balanced T
H
1- and T
H
2-based immune responses based on antibody isotyping. The triple mutant was cleared from mouse organs rapidly, with concurrent decreases in the production of various cytokines and histopathological lesions. When surviving animals infected with increasing doses of the triple mutant were subsequently challenged on day 24 with the bioluminescent WT CO92 strain (20 to 28 LD
50
s), 40 to 70% of the mice survived, with efficient clearing of the invading pathogen, as visualized in real time by
in vivo
imaging. The rapid clearance of the triple mutant, compared to that of WT CO92, from animals was related to the decreased adherence and invasion of human-derived HeLa and A549 alveolar epithelial cells and to its inability to survive intracellularly in these cells as well as in MH-S murine alveolar and primary human macrophages. An early burst of cytokine production in macrophages elicited by the triple mutant compared to WT CO92 and the mutant's sensitivity to the bactericidal effect of human serum would further augment bacterial clearance. Together, deletion of the
ail
gene from the Δ
lpp
Δ
msbB
double mutant severely attenuated
Y. pestis
CO92 to evoke pneumonic plague in a mouse model while retaining the required immunogenicity needed for subsequent protection against infection.
Competitive Science: Is Competition Ruining Science?
Authors: Fang Ferric C. , Casadevall Arturo 
Published: January 2015

ABSTRACTScience has always been a competitive undertaking. Despite recognition of the benefits of cooperation and team science, reduced availability of funding and jobs has made science more competitive than ever. Here we consider the benefits of competition in providing incentives to scientists and the adverse effects of competition on resource sharing, research integrity, and creativity. The history of science shows that transformative discoveries often occur in the absence of competition, which only emerges once fields are established and goals are defined. Measures to encourage collaboration and ameliorate competition in the scientific enterprise are discussed.
Burkholderia pseudomallei Type III Secretion System Cluster 3 ATPase BsaS, a Chemotherapeutic Target for Small-Molecule ATPase Inhibitors
Authors: Gong Lan , Lai Shu-Chin , Treerat Puthayalai , Prescott Mark , Adler Ben , Boyce John D. , Devenish Rodney J. 
Published: January 2015

ABSTRACT

Melioidosis is an infectious disease of high mortality for humans and other animal species; it is prevalent in tropical regions worldwide. The pathogenesis of melioidosis depends on the ability of its causative agent, the Gram-negative bacterium
Burkholderia pseudomallei
, to enter and survive in host cells.
B. pseudomallei
can escape from the phagosome into the cytosol of phagocytic cells where it replicates and acquires actin-mediated motility, avoiding killing by the autophagy-dependent process, LC3 (microtubule-associated protein light chain 3)-associated phagocytosis (LAP). The type III secretion system cluster 3 (TTSS3) facilitates bacterial escape from phagosomes, although the mechanism has not been fully elucidated. Given the recent identification of small-molecule inhibitors of the TTSS ATPase, we sought to determine the potential of the predicted TTSS3 ATPase, encoded by
bsaS
, as a target for chemotherapeutic treatment of infection. A
B. pseudomallei
bsaS
deletion mutant was generated and used as a control against which to assess the effect of inhibitor treatment. Infection of RAW 264.7 cells with wild-type bacteria and subsequent treatment with the ATPase inhibitor compound 939 resulted in reduced intracellular bacterial survival, reduced escape from phagosomes, and increased colocalization with both LC3 and the lysosomal marker LAMP1 (lysosome-associated membrane protein 1). These changes were similar to those observed for infection of RAW 264.7 cells with the
bsaS
deletion mutant. We propose that treatment with the ATPase inhibitor compound 939 decreased intracellular bacterial survival through a reduced ability of bacteria to escape from phagosomes and increased killing via LAP. Therefore, small-molecule inhibitors of the TTSS3 ATPase have potential as therapeutic treatments against melioidosis.
Engineering the Genome of Thermus thermophilus Using a Counterselectable Marker
Authors: Carr Jennifer F. , Danziger Michael E. , Huang Athena L. , Dahlberg Albert E. , Gregory Steven T. 
Published: January 2015

ABSTRACT

Thermus thermophilus
is an extremely thermophilic bacterium that is widely used as a model thermophile, in large part due to its amenability to genetic manipulation. Here we describe a system for the introduction of genomic point mutations or deletions using a counterselectable marker consisting of a conditionally lethal mutant allele of
pheS
encoding the phenylalanyl-tRNA synthetase α-subunit. Mutant PheS with an A294G amino acid substitution renders cells sensitive to the phenylalanine analog
p
-chlorophenylalanine. Insertion of the mutant
pheS
allele via a linked kanamycin resistance gene into a chromosomal locus provides a gene replacement intermediate that can be removed by homologous recombination using
p
-chlorophenylalanine as a counterselective agent. This selection is suitable for the sequential introduction of multiple mutations to produce a final strain unmarked by an antibiotic resistance gene. We demonstrated the utility of this method by constructing strains bearing either a point mutation in or a precise deletion of the
rrsB
gene encoding 16S rRNA. We also used this selection to identify spontaneous, large-scale deletions in the pTT27 megaplasmid, apparently mediated by either of the
T. thermophilus
insertion elements IS
Tth7
and IS
Tth8
. One such deletion removed 121 kb, including 118 genes, or over half of pTT27, including multiple sugar hydrolase genes, and facilitated the development of a plasmid-encoded reporter system based on β-galactosidase. The ability to introduce mutations ranging from single base substitutions to large-scale deletions provides a potentially powerful tool for engineering the genome of
T. thermophilus
and possibly other thermophiles as well.

IMPORTANCE
Thermus thermophilus
is an extreme thermophile that has played an important part in the development of both biotechnology and basic biological research. Its suitability as a genetic model system is established by its natural competence for transformation, but the scarcity of genetic tools limits the kinds of manipulations that can currently be performed. We have developed a counterselectable marker that allows the introduction of unmarked deletions and point mutations into the
T. thermophilus
genome. We find that this marker can also be used to select large chromosomal deletions apparently resulting from aberrant transposition of endogenous insertion sequences. This system has the potential to advance the genetic manipulation of this important model organism.
Evaluation of the Potential Impact of Ebola Virus Genomic Drift on the Efficacy of Sequence-Based Candidate Therapeutics
Authors: Kugelman Jeffrey R. , Sanchez-Lockhart Mariano , Andersen Kristian G. , Gire Stephen , Park Daniel J. , Sealfon Rachel , Lin Aaron E. , Wohl Shirlee , Sabeti Pardis C. , Kuhn Jens H. , Palacios Gustavo F. 
Published: January 2015

ABSTRACT
Until recently, Ebola virus (EBOV) was a rarely encountered human pathogen that caused disease among small populations with extraordinarily high lethality. At the end of 2013, EBOV initiated an unprecedented disease outbreak in West Africa that is still ongoing and has already caused thousands of deaths. Recent studies revealed the genomic changes this particular EBOV variant undergoes over time during human-to-human transmission. Here we highlight the genomic changes that might negatively impact the efficacy of currently available EBOV sequence-based candidate therapeutics, such as small interfering RNAs (siRNAs), phosphorodiamidate morpholino oligomers (PMOs), and antibodies. Ten of the observed mutations modify the sequence of the binding sites of monoclonal antibody (MAb) 13F6, MAb 1H3, MAb 6D8, MAb 13C6, and siRNA EK-1, VP24, and VP35 targets and might influence the binding efficacy of the sequence-based therapeutics, suggesting that their efficacy should be reevaluated against the currently circulating strain.
Work disability, lost productivity and associated risk factors in patients diagnosed with systemic lupus erythematosus
Authors: Utset Tammy O , Baskaran Amrutha , Segal Barbara M , Trupin Laura , Ogale Sarika , Herberich Ellen , Kalunian Kenneth 
Published: January 2015

ObjectiveTo assess prevalence and correlates of work presenteeism, absenteeism and work disability (WD) in patients with systemic lupus erythematous (SLE) and matched controls.MethodsPatients with SLE from six medical centres were recruited to complete a questionnaire consisting of several prevalidated survey instruments. The subject's rheumatologist completed medical history. Subjects recruited two non-SLE ‘best friend’ controls with matching demographics to complete a control survey. Analyses employed Student's t tests, χ2tests and logistic regression models.Results344 subjects with SLE and 322 controls submitted completed questionnaires. Mean pain, fatigue, Brief Cognitive Symptoms Index (BCSI) scores and depressive symptoms were worse in patients with SLE with WD (all p<0.01). WD was associated with African–American race, older age (51–65 years) and less than 4-year college education (all p<0.01). High presenteeism was associated with low pain and fatigue levels, higher BCSI scores and negatively correlated with depressive symptoms (all p<0.05). Increased pain and fatigue were associated with elevated absenteeism (p<0.05). Subjects with physically and cognitively demanding work reported worse presenteeism compared with controls with similar jobs (77% vs 85%, p<0.05 and 75% vs 85%, p<0.001), respectively. Patients with most cognitively demanding jobs reported greater weekly absenteeism (mean, 5.9 h) compared with controls (mean, 6.9 overtime hours, p<0.05).ConclusionsThe questionnaire demonstrated increased WD in SLE. Highly physical and highly cognitive jobs are challenging to patients with SLE and had increased absenteeism compared with controls. Depressive symptoms were correlated with better presenteeism without major socio-demographic determinants. Employability may be enhanced by improving treatment of depressive symptoms in patients with SLE.
Dietary Heterocyclic Amine Intake, NAT2 Genetic Polymorphism, and Colorectal Adenoma Risk: The Colorectal Adenoma Study in Tokyo
Authors: Budhathoki Sanjeev , Iwasaki Motoki , Yamaji Taiki , Sasazuki Shizuka , Takachi Ribeka , Sakamoto Hiromi , Yoshida Teruhiko , Tsugane Shoichiro 
Published: January 2015

Abstract
Background: While several studies have provided support for a positive association between meat intake and colorectal neoplasia, the role of heterocyclic amines (HCA), which is hypothesized to underline this relation, has been less consistent. We evaluated the association of HCA intake with colorectal adenoma risk in a case–control study in a middle-aged Japanese population.
Methods: Study subjects were 738 patients with adenoma and 697 controls who underwent total colonoscopy between 2004 and 2005 and responded to self-administered lifestyle and dietary questionnaires. HCA exposure concentration was estimated from meat and fish intake based on an HCA database that was validated against 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP) values measured in human hair. Logistic regression models were used to estimate ORs and 95% confidence interval (CI) for the association between HCA and colorectal adenoma risk after adjusting for potential confounders.
Results: High intake of 2-amino-3,4-dimethylimidazo[4,5-f]quinoline (MeIQ) and total HCA was associated with an increased risk of colorectal adenoma in women but not in men. The multivariate-adjusted OR for the highest versus lowest quartile in women was 2.10 (95% CI, 1.20–3.67; Ptrend = 0.01) for MeIQ and 1.73 (95% CI, 0.99–3.01; Ptrend = 0.03) for total HCA. No clear association with PhIP or 2-amino-3,8-dimethylimidazo[4,5-f]quinoxaline (MeIQx) estimates and no effect modification by NAT2 acetylation genotype was observed.
Conclusions: This study suggests that high MeIQ and total HCA estimates are positively associated with colorectal adenoma risk.
Impact: The findings add to evidence that HCA may play a role in colorectal carcinogenesis in humans. Cancer Epidemiol Biomarkers Prev; 24(3); 613–20. ©2015 AACR.