bluuebunny/crossref_metadata_2025 huggingface dataset:
Inclusion of the Bovine Neutrophil Beta-Defensin 3 with Glycoprotein D of Bovine Herpesvirus 1 in a DNA Vaccine Modulates Immune Responses of Mice and Cattle
Authors: Mackenzie-Dyck Sarah , Kovacs-Nolan Jennifer , Snider Marlene , Babiuk Lorne A. , van Drunen Littel-van den Hurk Sylvia
Published: January 2014

Bovine herpesvirus 1 (BoHV-1) causes recurrent respiratory and genital infections in cattle and predisposes them to lethal secondary infections. While modified live and killed BoHV-1 vaccines exist, these are not without problems. Development of an effective DNA vaccine for BoHV-1 has the potential to address these issues. As a strategy to enhance DNA vaccine immunity, a plasmid encoding the bovine neutrophil beta-defensin 3 (BNBD3) as a fusion with truncated glycoprotein D (tgD) and a mix of two plasmids encoding BNBD3 and tgD were tested in mice and cattle. In mice, coadministration of BNBD3 on the separate plasmid enhanced the tgD-induced gamma interferon (IFN-γ) response but not the antibody response. BNBD3 fused to tgD did not affect the antibody levels or the number of IFN-γ-secreting cells but increased the induction of tgD-specific cytotoxic T lymphocytes (CTLs). In cattle, the addition of BNBD3 as a fusion construct also modified the immune response. While the IgG and virus-neutralizing antibody levels were not affected, the number of IFN-γ-secreting cells was increased after BoHV-1 challenge, specifically the CD8+IFN-γ+T cells, including CD8+IFN-γ+CD25+CTLs. While reduced virus shedding, rectal temperature, and weight loss were observed, the level of protection was comparable to that observed in pMASIA-tgD-vaccinated animals. These data show that coadministration of BNBD3 with a protective antigen as a fusion in a DNA vaccine strengthened the Th1 bias and increased cell-mediated immune responses but did not enhance protection from BoHV-1 infection.
Purification, Stability, and Immunogenicity Analyses of Five Bluetongue Virus Proteins for Use in Development of a Subunit Vaccine That Allows Differentiation of Infected from Vaccinated Animals
Authors: Anderson Jenna , Bréard Emmanuel , Lövgren Bengtsson Karin , Grönvik Kjell-Olov , Zientara Stéphan , Valarcher Jean-Francois , Hägglund Sara
Published: January 2014

Bluetongue virus (BTV) causes bluetongue disease, a vector-borne disease of ruminants. The recent northerly spread of BTV serotype 8 in Europe resulted in outbreaks characterized by clinical signs in cattle, including unusual teratogenic effects. Vaccination has been shown to be crucial for controlling the spread of vector-borne diseases such as BTV. With the aim of developing a novel subunit vaccine targeting BTV-8 that allows differentiation of infected from vaccinated animals, five His-tagged recombinant proteins, VP2 and VP5 of BTV-8 and NS1, NS2, and NS3 of BTV-2, were expressed in baculovirus orEscherichia coliexpression systems for further study. Optimized purification protocols were determined for VP2, NS1, NS2, and NS3, which remained stable for detection for at least 560 to 610 days of storage at +4°C or −80°C, and Western blotting using sera from vaccinated or experimentally infected cattle indicated that VP2 and NS2 were recognized by BTV-specific antibodies. To characterize murine immune responses to the four proteins, mice were subcutaneously immunized twice at a 4-week interval with one of three protein combinations plus immunostimulating complex ISCOM-Matrix adjuvant or with ISCOM-Matrix alone (n= 6 per group). Significantly higher serum IgG antibody titers specific for VP2 and NS2 were detected in immunized mice than were detected in controls. VP2, NS1, and NS2 but not NS3 induced specific lymphocyte proliferative responses upon restimulation of spleen cells from immunized mice. The data suggest that these recombinant purified proteins, VP2, NS1, and NS2, could be an important part of a novel vaccine design against BTV-8.
Phosphatidylinositol-Specific Phospholipase C Contributes to Survival of Staphylococcus aureus USA300 in Human Blood and Neutrophils
Authors: White Mark J. , Boyd Jeffrey M. , Horswill Alexander R. , Nauseef William M.
Published: January 2014

Staphylococcus aureus
is an important human pathogen that employs a large repertoire of secreted virulence factors to promote disease pathogenesis. Many strains of
S. aureus
possess a
plc
gene that encodes a phosphatidylinositol (PI)-specific phospholipase C (PI-PLC) capable of hydrolyzing PI and cleaving glycosyl-PI (GPI)-linked proteins from cell surfaces. Despite being secreted by virulent staphylococci, the contribution of PI-PLC to the capacity of
S. aureus
to cause disease remains undefined. Our goal in these studies was to understand PI-PLC in the context of
S. aureus
biology. Among a collection of genetically diverse clinical isolates of
S. aureus
, community-associated methicillin-resistant
S. aureus
(CA-MRSA) USA300 secreted the most PI-PLC. Screening a collection of two-component system (TCS) mutants of
S. aureus
, we identified both the
agr
quorum-sensing system and the SrrAB TCS to be positive regulators of
plc
gene expression. Real-time PCR and PI-PLC enzyme assays of the TCS mutants, coupled with SrrA promoter binding studies, demonstrated that SrrAB was the predominant transcriptional activator of
plc
. Furthermore,
plc
regulation was linked to oxidative stress both
in vitro
and
in vivo
in a SrrAB-dependent manner. A Δ
plc
mutant in a CA-MRSA USA300 background exhibited a survival defect in human whole blood and in isolated neutrophils. However, the same mutant strain displayed no survival defect in murine models of infection or murine whole blood. Overall, these data identify potential links between bacterial responses to the host innate immune system and to oxidative stress and suggest how PI-PLC could contribute to the pathogenesis of
S. aureus
infections.
Incorporation of Phosphorylcholine into the Lipooligosaccharide of Nontypeable Haemophilus influenzae Does Not Correlate with the Level of Biofilm Formation
In Vitro
Authors: Puig Carmen , Marti Sara , Hermans Peter W. M. , de Jonge Marien I. , Ardanuy Carmen , Liñares Josefina , Langereis Jeroen D.
Published: January 2014

Nontypeable
Haemophilus influenzae
(NTHi) is an opportunistic pathogen that causes otitis media in children and community-acquired pneumonia or exacerbations of chronic obstructive pulmonary disease in adults. A large variety of studies suggest that biofilm formation by NTHi may be an important step in the pathogenesis of this bacterium. The objective of this report was to determine the relationship between the presence of phosphorylcholine in the lipooligosaccharide of NTHi and the level of biofilm formation. The study was performed on 111 NTHi clinical isolates collected from oropharyngeal samples of healthy children, middle ear fluid of children with otitis media, and sputum samples of patients with chronic obstructive pulmonary disease or community-acquired pneumonia. NTHi clinical isolates presented a large variation in the level of biofilm formation in a static assay and phosphorylcholine content. Isolates collected from the oropharynx and middle ear fluid of children tended to have more phosphorylcholine and made denser biofilms than isolates collected from sputum samples of patients with chronic obstructive pulmonary disease or community-acquired pneumonia. No correlation was observed between biofilm formation and the presence of phosphorylcholine in the lipooligosaccharide for either planktonic or biofilm growth. This lack of correlation was confirmed by abrogating phosphorylcholine incorporation into lipooligosaccharide through
licA
gene deletion, which had strain-specific effects on biofilm formation. Altogether, we present strong evidence to conclude that there is no correlation between biofilm formation in a static assay and the presence of phosphorylcholine in lipooligosaccharide in a large collection of clinical NTHi isolates collected from different groups of patients.
Neutrophils Select Hypervirulent CovRS Mutants of M1T1 Group A Streptococcus during Subcutaneous Infection of Mice
Authors: Li Jinquan , Liu Guanghui , Feng Wenchao , Zhou Yang , Liu Mengyao , Wiley James A. , Lei Benfang
Published: January 2014

Pathogen mutants arise during infections. Mechanisms of selection for pathogen variants are poorly understood. We tested whether neutrophils select mutations in the two-component regulatory system CovRS of group A
Streptococcus
(GAS) during infection using the lack of production of the protease SpeB (SpeB activity negative [SpeB
A−
]) as a marker. Depletion of neutrophils by antibodies RB6-8C5 and 1A8 reduced the percentage of SpeB
A−
variants (SpeB
A−
%) recovered from mice infected with GAS strain MGAS2221 by >76%. Neutrophil recruitment and SpeB
A−
% among recovered GAS were reduced by 95% and 92%, respectively, in subcutaneous MGAS2221 infection of CXCR2
−/−
mice compared with control mice. In air sac infection with MGAS2221, levels of neutrophils and macrophages in lavage fluid were reduced by 49% and increased by 287%, respectively, in CXCR2
−/−
mice compared with control mice, implying that macrophages play an insignificant role in the reduction of selection for SpeB
A−
variants in CXCR2
−/−
mice. One randomly chosen SpeB
A−
mutant outcompeted MGAS2221 in normal mice but was outcompeted by MGAS2221 in neutropenic mice and had enhancements in expression of virulence factors, innate immune evasion, skin invasion, and virulence. This and nine other SpeB
A−
variants from a mouse all had nonsynonymous
covRS
mutations that resulted in the SpeB
A−
phenotype and enhanced expression of the CovRS-controlled secreted streptococcal esterase (SsE). Our findings are consistent with a model that neutrophils select spontaneous
covRS
mutations that maximize the potential of GAS to evade neutrophil responses, resulting in variants with enhanced survival and virulence. To our knowledge, this is the first report of the critical contribution of neutrophils to the selection of pathogen variants.
Identification of Point Mutations in Clinical Staphylococcus aureus Strains That Produce Small-Colony Variants Auxotrophic for Menadione
Authors: Dean Melissa A. , Olsen Randall J. , Long S. Wesley , Rosato Adriana E. , Musser James M.
Published: January 2014

Staphylococcus aureus
small-colony variants (SCVs) are implicated in chronic and relapsing infections that are difficult to diagnose and treat. Despite many years of study, the underlying molecular mechanisms and virulence effect of the small-colony phenotype remain incompletely understood. We sequenced the genomes of five
S. aureus
SCV strains recovered from human patients and discovered previously unidentified nonsynonymous point mutations in three genes encoding proteins in the menadione biosynthesis pathway. Analysis of genetic revertants and complementation with wild-type alleles confirmed that these mutations caused the SCV phenotype and decreased virulence for mice.
CsrA (BB0184) Is Not Involved in Activation of the RpoN-RpoS Regulatory Pathway in Borrelia burgdorferi
Authors: Ouyang Zhiming , Zhou Jianli , Norgard Michael V.
Published: January 2014

Borrelia burgdorferi
encodes a homologue of the bacterial carbon storage regulator A (CsrA). Recently, it was reported that CsrA contributes to
B. burgdorferi
infectivity and is required for the activation of the central RpoN-RpoS regulatory pathway. However, many questions concerning the function of CsrA in
B. burgdorferi
gene regulation remain unanswered. In particular, there are conflicting reports concerning the molecular details of how CsrA may modulate
rpoS
expression and, thus, how CsrA may influence the RpoN-RpoS pathway in
B. burgdorferi
. To address these key discrepancies, we examined the role of CsrA in differential gene expression in the Lyme disease spirochete. Upon engineering an inducible
csrA
expression system in
B. burgdorferi
, controlled hyperexpression of CsrA in a merodiploid strain did not significantly alter the protein and transcript levels of
bosR
,
rpoS
, and RpoS-dependent genes (such as
ospC
and
dbpA
). In addition, we constructed isogenic
csrA
mutants in two widely used infectious
B. burgdorferi
strains. When expression of
bosR
,
rpoS
,
ospC
, and
dbpA
was compared between the
csrA
mutants and their wild-type counterparts, no detectable differences were observed. Finally, animal studies indicated that the
csrA
mutants remained infectious for and virulent in mice. Analyses of
B. burgdorferi
gene expression in mouse tissues showed comparable levels of
rpoS
transcripts by the
csrA
mutants and the parental strains. Taken together, these results constitute compelling evidence that CsrA is not involved in activation of the RpoN-RpoS pathway and is dispensable for mammalian infectious processes carried out by
B. burgdorferi
.
Induction of Virulence Gene Expression in Staphylococcus aureus by Pulmonary Surfactant
Authors: Ishii Kenichi , Adachi Tatsuo , Yasukawa Jyunichiro , Suzuki Yutaka , Hamamoto Hiroshi , Sekimizu Kazuhisa
Published: January 2014

We performed a genomewide analysis using a next-generation sequencer to investigate the effect of pulmonary surfactant on gene expression in
Staphylococcus aureus
, a clinically important opportunistic pathogen. RNA sequence (RNA-seq) analysis of bacterial transcripts at late log phase revealed 142 genes that were upregulated >2-fold following the addition of pulmonary surfactant to the culture medium. Among these genes, we confirmed by quantitative reverse transcription-PCR analysis that mRNA amounts for genes encoding ESAT-6 secretion system C (EssC), an unknown hypothetical protein (NWMN_0246; also called pulmonary surfactant-inducible factor A [PsiA] in this study), and hemolysin gamma subunit B (HlgB) were increased 3- to 10-fold by the surfactant treatment. Among the major constituents of pulmonary surfactant, i.e., phospholipids and palmitate, only palmitate, which is the most abundant fatty acid in the pulmonary surfactant and a known antibacterial substance, stimulated the expression of these three genes. Moreover, these genes were also induced by supplementing the culture with detergents. The induction of gene expression by surfactant or palmitate was not observed in a disruption mutant of the
sigB
gene, which encodes an alternative sigma factor involved in bacterial stress responses. Furthermore, each disruption mutant of the
essC
,
psiA
, and
hlgB
genes showed attenuation of both survival in the lung and host-killing ability in a murine pneumonia model. These findings suggest that
S. aureus
resists membrane stress caused by free fatty acids present in the pulmonary surfactant through the regulation of virulence gene expression, which contributes to its pathogenesis within the lungs of the host animal.
Genomic Portrait of the Evolution and Epidemic Spread of a Recently Emerged Multidrug-Resistant Shigella flexneri Clone in China
Authors: Zhang Nan , Lan Ruiting , Sun Qiangzheng , Wang Jianping , Wang Yiting , Zhang Jin , Yu Deshan , Hu Wanfu , Hu Shoukui , Dai Hang , Du Pengcheng , Wang Haiyin , Xu Jianguo
Published: January 2014

Shigella flexneri
is the major cause of shigellosis in developing countries. A new
S. flexneri
serotype, Xv, appeared in 2000 and replaced serotype 2a as the most prevalent serotype in China. Serotype Xv is a variant of serotype X, with phosphoethanolamine modification of its O antigen mediated by a plasmid that contained the
opt
gene. Serotype Xv isolates belong to sequence type 91 (ST91). In this study, whole-genome sequencing of 59
S. flexneri
isolates of 14 serotypes (serotypes 1 to 4, Y, Yv, X, and Xv) indicated that ST91 arose around 1993 by acquiring multidrug resistance (MDR) and spread across China within a decade. A comparative analysis of the chromosome and
opt
-carrying plasmid pSFXv_2 revealed independent origins of 3 serotype Xv clusters in China, with different divergence times. Using 18 cluster-dividing single-nucleotide polymorphisms (SNPs), SNP typing divided 380 isolates from 3 provinces (Henan, Gansu, and Anhui) into 5 SNP genotypes (SGs). One SG predominated in each province, but substantial interregional spread of SGs was also evident. These findings suggest that MDR is the key selective pressure for the emergence of the
S. flexneri
epidemic clone and that
Shigella
epidemics in China were caused by a combination of local expansion and interregional spread of serotype Xv.
Detection of Campylobacter in Stool and Determination of Significance by Culture, Enzyme Immunoassay, and PCR in Developing Countries
Authors: Platts-Mills James A. , Liu Jie , Gratz Jean , Mduma Esto , Amour Caroline , Swai Ndealilia , Taniuchi Mami , Begum Sharmin , Peñataro Yori Pablo , Tilley Drake H. , Lee Gwenyth , Shen Zeli , Whary Mark T. , Fox James G. , McGrath Monica , Kosek Margaret , Haque Rashidul , Houpt Eric R.
Published: January 2014

Campylobacter
is a common bacterial enteropathogen that can be detected in stool by culture, enzyme immunoassay (EIA), or PCR. We compared culture for
C. jejuni/C. coli
, EIA (ProSpecT), and duplex PCR to distinguish
Campylobacter jejuni/C. coli
and non-
jejuni/coli Campylobacter
on 432 diarrheal and matched control stool samples from infants in a multisite longitudinal study of enteric infections in Tanzania, Bangladesh, and Peru. The sensitivity and specificity of culture were 8.5% and 97.6%, respectively, compared with the results of EIA and 8.7% and 98.0%, respectively, compared with the results of PCR for
C. jejuni/C. coli
. Most (71.6%) EIA-positive samples were positive by PCR for
C. jejuni/C. coli
, but 27.6% were positive for non-
jejuni/coli Campylobacter
species. Sequencing of 16S rRNA from 53 of these non-
jejuni/coli Campylobacter
samples showed that it most closely matched the 16S rRNA of
C. hyointestinalis
subsp.
lawsonii
(56%),
C. troglodytis
(33%),
C. upsaliensis
(7.7%), and
C. jejuni/C. coli
(2.6%).
Campylobacter
-negative stool spiked with each of the above-mentioned
Campylobacter
species revealed reactivity with EIA. PCR detection of
Campylobacter
species was strongly associated with diarrhea in Peru (odds ratio [OR] = 3.66,
P
< 0.001) but not in Tanzania (OR = 1.56,
P
= 0.24) or Bangladesh (OR = 1.13,
P
= 0.75). According to PCR,
Campylobacter jejuni/C. coli
infections represented less than half of all infections with
Campylobacter
species. In sum, in infants in developing country settings, the ProSpecT EIA and PCR for
Campylobacter
reveal extremely high rates of positivity. We propose the use of PCR because it retains high sensitivity, can ascertain burden, and can distinguish between
Campylobacter
infections at the species level.
Concordance of PCR and Culture from Nasal Swabs for Detection of Methicillin-Resistant Staphylococcus aureus in a Setting of Concurrent Antistaphylococcal Antibiotics
Authors: Shenoy Erica S. , Noubary Farzad , Kim JiYeon , Rosenberg Eric S. , Cotter Jessica A. , Lee Hang , Walensky Rochelle P. , Hooper David C.
Published: January 2014

The effect of concurrent administration of antibiotics on the detection of methicillin-resistant
Staphylococcus aureus
(MRSA) remains unresolved. Here, we assessed the concordance of paired nasal swabs processed using commercial PCR and culture and found high concordance in both the absence and presence of antibiotics with activity against MRSA (93.7% [95% confidence interval [CI], 88.1%, 96.8%] and 90.9% [95% CI, 84.8%, 94.7%], respectively), although PCR was more likely to be positive in the presence of antibiotics. (This study has been registered at
ClinicalTrials.gov
under registration no. NCT01234831.)
Phenotypic and Genotypic Characterization of Canadian Clinical Isolates of Vibrio parahaemolyticus Collected from 2000 to 2009
Authors: Banerjee Swapan K. , Kearney Ashley K. , Nadon Celine A. , Peterson Christy-Lynn , Tyler Kevin , Bakouche Laurene , Clark Clifford G. , Hoang Linda , Gilmour Matthew W. , Farber Jeffrey M.
Published: January 2014

Vibrio parahaemolyticus
is the leading bacterial cause of food-borne illness due to the consumption of contaminated seafood. The aim of the present study was to determine the population of its subtypes and establish a better understanding of the various types of
V. parahaemolyticus
strains that are causing human illness in Canada. The subtypes for 100 human clinical isolates of
V. parahaemolyticus
collected between 2000 and 2009 were determined by performing serotyping, ribotyping, pulsed-field gel electrophoresis, and multilocus sequence typing. Within this panel of strains, there was a high level of diversity (between 22 and 53 subtypes per method), but the presence of predominant clones with congruent subtypes between the various methods was also observed. For example, all 32 isolates belonging to sequence type 36 (ST36) were from serogroup O4, while 31 of them were ribotype EcoVib235-287, and 24 of the 32 were SfiI pulsed-field gel electrophoresis (PFGE) pattern VPSF1.0001. With regard to the presence of known virulence genes, 74 of the 100 isolates were PCR positive for the presence of the thermostable direct hemolysin (
tdh
); and 59 of these 74 strains also contained the second virulence marker, the
tdh
-related hemolysin (
trh
). The detection of
trh
was more predominant (81%) among the clinical isolates, and only four (4%) of the clinical isolates tested negative for the presence of both
tdh
and
trh
. This database, comprising 100 clinical isolates of
V. parahaemolyticus
strains from Canada, forms a baseline understanding of subtype diversity for future source attribution and other epidemiologic studies.
Association of Herpes Simplex Virus pU
L
31 with Capsid Vertices and Components of the Capsid Vertex-Specific Complex
Authors: Yang Kui , Wills Elizabeth , Lim Han Young , Zhou Z. Hong , Baines Joel D.
Published: January 2014

pU
L
34 and pU
L
31 of herpes simplex virus (HSV) comprise the nuclear egress complex (NEC) and are required for budding at the inner nuclear membrane. pU
L
31 also associates with capsids, suggesting it bridges the capsid and pU
L
34 in the nuclear membrane to initiate budding. Previous studies showed that capsid association of pU
L
31 was precluded in the absence of the C terminus of pU
L
25, which along with pU
L
17 comprises the capsid vertex-specific complex, or CVSC. The present studies show that the final 20 amino acids of pU
L
25 are required for pU
L
31 capsid association. Unexpectedly, in the complete absence of pU
L
25, or when pU
L
25 capsid binding was precluded by deletion of its first 50 amino acids, pU
L
31 still associated with capsids. Under these conditions, pU
L
31 was shown to coimmunoprecipitate weakly with pU
L
17. Based on these data, we hypothesize that the final 20 amino acids of pU
L
25 are required for pU
L
31 to associate with capsids. In the absence of pU
L
25 from the capsid, regions of capsid-associated pU
L
17 are bound by pU
L
31. Immunogold electron microscopy revealed that pU
L
31 could associate with multiple sites on a single capsid in the nucleus of infected cells. Electron tomography revealed that immunogold particles specific to pU
L
31 protein bind to densities at the vertices of the capsid, a location consistent with that of the CVSC. These data suggest that pU
L
31 loads onto CVSCs in the nucleus to eventually bind pU
L
34 located within the nuclear membrane to initiate capsid budding.

IMPORTANCE
This study is important because it localizes pU
L
31, a component previously known to be required for HSV capsids to bud through the inner nuclear membrane, to the vertex-specific complex of HSV capsids, which comprises the unique long region 25 (U
L
25) and U
L
17 gene products. It also shows this interaction is dependent on the C terminus of U
L
25. This information is vital for understanding how capsids bud through the inner nuclear membrane.
Evidence of
In Utero
Transmission of Classical Scrapie in Sheep
Authors: Spiropoulos John , Hawkins Stephen A. C. , Simmons Marion M. , Bellworthy Susan J.
Published: January 2014

Classical scrapie is one of the transmissible spongiform encephalopathies (TSEs), a group of fatal infectious diseases that affect the central nervous system (CNS). Classical scrapie can transmit laterally from ewe to lamb perinatally or between adult animals. Here we report detection of infectivity in tissues of an unborn fetus, providing evidence that
in utero
transmission of classical scrapie is also possible.
Relocation of the NIb Gene in the Tobacco Etch Potyvirus Genome
Authors: Majer Eszter , Salvador Zaira , Zwart Mark P. , Willemsen Anouk , Elena Santiago F. , Daròs José-Antonio
Published: January 2014

Potyviruses express most of their proteins from a long open reading frame that is translated into a large polyprotein processed by three viral proteases. To understand the constraints on potyvirus genome organization, we relocated the viral RNA-dependent RNA polymerase (NIb) cistron to all possible intercistronic positions of the
Tobacco etch virus
(TEV) polyprotein. Only viruses with NIb at the amino terminus of the polyprotein or in between P1 and HC-Pro were viable in tobacco plants.
Stability of the Parainfluenza Virus 5 Genome Revealed by Deep Sequencing of Strains Isolated from Different Hosts and following Passage in Cell Culture
Authors: Rima Bert K. , Gatherer Derek , Young Daniel F. , Norsted Hanna , Randall Richard E. , Davison Andrew J.
Published: January 2014

The strain diversity of a rubulavirus, parainfluenza virus 5 (PIV5), was investigated by comparing 11 newly determined and 6 previously published genome sequences. These sequences represent 15 PIV5 strains, of which 6 were isolated from humans, 1 was from monkeys, 2 were from pigs, and 6 were from dogs. Strain diversity is remarkably low, regardless of host, year of isolation, or geographical origin; a total of 7.8% of nucleotides are variable, and the average pairwise difference between strains is 2.1%. Variation is distributed unevenly across the PIV5 genome, but no convincing evidence of selection for antibody-mediated evasion in hemagglutinin-neuraminidase was found. The finding that some canine and porcine, but not primate, strains are mutated in the SH gene, and do not produce SH, raised the possibility that dogs (or pigs) may not be the natural host of PIV5. The genetic stability of PIV5 was also demonstrated during serial passage of one strain (W3) in Vero cells at a high multiplicity of infection, under conditions of competition with large proportions of defective interfering genomes. A similar observation was made for a strain W3 mutant (PIV5VΔC) lacking V gene function, in which the dominant changes were related to pseudoreversion in this gene. The mutations detected in PIV5VΔC during pseudoreversion, and also those characterizing the SH gene in canine and porcine strains, predominantly involved U-to-C transitions. This suggests an important role for biased hypermutation via an adenosine deaminase, RNA-specific (ADAR)-like activity.

IMPORTANCE
Here we report the sequence variation of 16 different isolates of parainfluenza virus 5 (PIV5) that were isolated from a number of species, including humans, monkeys, dogs, and pigs, over 4 decades. Surprisingly, strain diversity was remarkably low, regardless of host, year of isolation, or geographical origin. Variation was distributed unevenly across the PIV5 genome, but no convincing evidence of immune or host selection was found. This overall genome stability of PIV5 was also observed when the virus was grown in the laboratory, and the genome stayed remarkably constant even during the selection of virus mutants. Some of the canine isolates had lost their ability to encode one of the viral proteins, termed SH, suggesting that although PIV5 commonly infects dogs, dogs may not be the natural host for PIV5.
Caecal volvulus following abdominoplasty: a tale of caution
Authors: Zakeri Simon Arian , Hotouras Alex , Mukhtar Hasan , Bhan Chetan
Published: January 2014

We present a case of a 30-year-old woman with a gangrenous right colon secondary to caecal volvulus, diagnosed postabdominoplasty. To our knowledge, this is the first case recorded in the literature. An emergency right hemicolectomy was performed and the patient had an unremarkable recovery. Given the severity of this incident, the diagnosis of caecal volvulus postabdominoplasty requires a high index of suspicion to allow prompt diagnosis and treatment.
Siderosis bulbi as a consequence of a missed intraocular foreign body
Authors: Lapira Matthew , Karl David , Murgatroyd Helen
Published: January 2014

We present a case of a 56-year-old man who suffered an injury to his right eye in June 2012. He presented to an emergency department, however, the presence of a penetrating injury and an intraocular foreign body was not identified. A year later he was referred to the ophthalmology department due to reduced vision and change of iris colour in the same eye. Examination revealed clinical signs consistent with a previous penetrating injury and a retained ferrous intraocular foreign body.
Caring at the end of life: do cancer caregivers differ from other caregivers?
Authors: Girgis Afaf , Abernethy Amy P , Currow David C
Published: January 2014

Objective

Cancer is one of the most common health conditions in receipt of informal caregiving. This study compares key characteristics of caregivers who cared for someone with cancer until death with caregivers of people with other life-limiting illnesses and their care recipients irrespective of health service utilisation.

Method

Data were analysed from annual state-wide South Australian Health Omnibus Surveys (2000–2007) involving 14 624 respondents, regarding end of life care. Descriptive and comparative data are presented.

Results

Almost a third of respondents (32%; participation rate 72%) had someone close to them die from an ‘expected’ death in the preceding 5 years. One in 10 (10%) respondents reported providing hands-on care predominantly for someone with cancer. Compared with non-cancer caregivers, cancer caregivers cared for someone who was significantly younger (mean age 66 (95% CI 64 to 67) years vs 74 (95% CI 72 to 77) years; one-way analysis of variance p<0.0001) and were more likely to report having a hospice/palliative care service involved in the care of the deceased (65% (95% CI 63 to 67) compared with 39% (95% CI 37 to 42). In the Australian context, this may mean contact with inpatient, outpatient and community-based services.

There were no differences between the needs which caregivers perceived to be unmet or the perceptions that no additional supports were required between the two groups.

Conclusions

Informal caregivers perform a critical social and economic role in care provision. Cancer caregivers are a proportionally larger cohort than non-cancer caregivers. With the increasing incidence of cancer, the sustainability of a voluntary cancer caregiving workforce will be reliant upon minimising the burden of care.
Superior outcomes for military ankylosing spondylitis patients treated with anti-TNF
Authors: Rees Jonathan D , Bennett A N , Harris D , Jones T
Published: January 2014

Objectives

The British military has a cohort of patients with ankylosing spondylitis (AS) characterised by young age and short disease duration. Many of the most severely affected AS patients have been treated since 2005 at Headley Court with anti-tumour necrosis factor (anti-TNF) therapy in accordance with National Institute of Health and Care Excellence guidance. We wanted to prospectively determine both the safety and efficacy of this new treatment in our British military population and compare this with relevant civilian study outcome data.

Methods

All AS patients commenced on anti-TNF therapy at Headley Court were prospectively monitored for treatment efficacy and side effects. Outcome measures used included the Bath Ankylosing Spondylitis Disease Activity Index. Our results were compared with a civilian comparison group (NHS) and relevant landmark clinical trial data.

Results

Our patients were younger (mean age 34.7 years) and had a shorter duration (mean disease duration 6.9 years) than the civilian (NHS) comparison group. Our safety data were extremely benign with only two patients suffering minor side effects (local injection site reaction). Furthermore, our outcome data were superior to both NHS routine care and to landmark clinical studies.

Discussion

Prior to this study, there were no data on military AS populations receiving anti-TNF therapy. The study confirms British military patients tolerate this therapy extremely well and receive greater benefit from this treatment than that seen in any published study to date. We believe that this confirms that young age and short disease duration are good prognostic factors in the treatment of AS with anti-TNF therapy.
Synthetic studies on heterocyclic natural products
Authors: Ciufolini Marco A.
Published: January 2014

This paper highlights ongoing efforts toward Erythrina alkaloids, himandrine, tetrodotoxin, and thiopeptide antibiotics such as nosiheptide and describes representative spinoffs in biomedicine that emanated from the author’s research in synthetic organic chemistry.
Hydraulics of backflushing for efficiently cleaning hydropower trash racks
Authors: Nøvik Hanne , Lia Leif , Rettedal Bjørnar , Nielsen Lars Eid
Published: January 2014

Backflushing is an efficient alternative for cleaning trash racks at small hydropower plants. The actual debris to trash rack adhesion is measured in situ for the first time by an innovative, custom-made measuring device. Despite large measurement variations caused by unevenness in debris distribution, the adhesion to trash rack for most types of debris and for most sites is relatively modest. There is also a trend where a higher maximum head loss through the trash rack increases the debris adhesion to the trash rack. The hydraulics around the trash rack during a backflushing process is assessed by monitoring the velocity development and the pressure difference through the trash rack in a physical model. As expected, when debris adhesion to the trash rack is high, releasing the attached debris requires a higher pressure difference through the trash rack and a higher approach velocity.
Bootstrapping of sample sizes for length- or age-composition data used in stock assessments
Authors: Stewart Ian J. , Hamel Owen S.
Published: January 2014

Integrated stock assessment models derive estimates of management quantities by fitting to indices of abundance and length and age compositions. For composition data, where a multinomial likelihood is often applied, weights are determined by input sample sizes, which can be an important contributor to model results. We used a generic bootstrap method, verified through simulation, to calculate year-specific maximum realized sample sizes from the observation error inherent in fishery biological data. Applying this method to length-composition observations for 47 groundfish species collected during a standardized trawl survey, we found maximum realized sample size to be related to both the number of hauls and individual fish sampled from those hauls. Sampling in excess of 20 fish from each haul produced little increase in most cases, with maximum realized sample size ranging from approximately 2 to 4 per haul sampled. Utilizing these maximum realized sample sizes as input values for stock assessment (analogous to minimum variance estimates) appropriately incorporates interannual variability, and may reduce over-emphasis on composition data. Results from this method can also help determine sampling targets.
Wood density of Norway spruce in uneven-aged stands
Authors: Piispanen Riikka , Heinonen Jaakko , Valkonen Sauli , Mäkinen Harri , Lundqvist Sven-Olof , Saranpää Pekka
Published: January 2014

This study focused on wood density and annual ring width in Norway spruce (Picea abies (L.) Karst.) grown in uneven-aged stands (UAS). In total, 96 trees were harvested from five UAS that had been managed by single-tree selection for decades. A data set of 27 trees from even-aged stands (EAS) was used for comparison. In the UAS trees, high density and narrow annual rings were found in the juvenile wood near the pith. Thereafter, wood density rapidly decreased until the 20th annual ring, followed by an increase toward the bark. In the outermost rings, wood density again slightly decreased. The trends in wood density in the UAS trees correspond with those reported for naturally regenerated, even-aged Norway spruce stands, with the exception of the decrease in the outermost rings. A mixed linear model with ring width, cambial age, and canopy position as fixed parameters accounted for 53% of the variation in wood density of the UAS trees. In contrast to UAS trees, EAS trees showed increased wood density in the outer rings as a result of decreasing growth rate. The abrupt change in wood density of UAS trees may affect the properties of timber sawn close to the pith.
Facultative nocturnal behaviour in snakes: experimental examination of why and how with Ratsnakes (Elaphe obsoleta) and Racers (Coluber constrictor)
Authors: DeGregorio Brett A. , Sperry Jinelle H. , Valente Daniel P. , Weatherhead Patrick J.
Published: January 2014

Diel activity patterns are often fixed within species such that most animals can be classified as diurnal, crepuscular, or nocturnal, and have sensory abilities that reflect when they are active. However, many snake species appear capable of switching between diurnal and nocturnal activity. Here, we evaluate the hypothesis that some species are constrained in their activity by the sensory cues used for foraging. We experimentally assessed differences between two sympatric snake species in their ability to alter diel activity patterns, to address why those snakes that switch do so (do thermal constraints force them to be active in otherwise nonpreferred conditions?), and to explore how sensory abilities to locate prey facilitate or constrain this shift. Ratsnakes (Elaphe obsoleta (Say in James, 1823)) were active when temperature was optimal, regardless of light level, suggesting their activity pattern is genuinely plastic. Consistent with our predictions, Ratsnakes successfully detected prey in low and high light using visual or chemical cues, and were most successful when cues were coupled. Racers (Coluber constrictor L., 1758) were almost exclusively diurnal, regardless of temperature, and became less active when daytime temperatures were suboptimal. The ability of Ratsnakes to shift activity may confer a foraging advantage and should buffer Ratsnakes and similarly flexible species from climate change, whereas climate change may pose a more serious threat to inflexible species such as Racers.
STUDY OF PARASITIC CAPACITANCE EFFECT ON NOISE FIGURE OF IDCLNA IN DEEP SUBMICRON CMOS TECHNOLOGIES
Authors: MANJULA S. , SELVATHI D.
Published: January 2014

Low noise amplifier (LNA) is an important component in RF receiver front end. An inductively degenerated cascode low noise amplifier (IDCLNA) is mostly preferred for producing good trade-offs such as high gain, low noise figure (NF), high reverse isolation and low power consumption for narrowband applications. This IDCLNA structure is also used to reduce the gate induced noise on the noise performance by inserting the capacitance in parallel with the gate-to-source capacitance of main transistor. Usually, the parasitic overlap capacitances can impose serious constraints on achievable performance and is taken into account in IDCLNA. In this paper, IDCLNA is designed at a frequency of 2.4 GHz with analyzing the impact of parasitic overlap capacitances on IDCLNA in terms of unity current gain frequency (f T ) which will affect the NF of IDCLNA and simulated using 130 nm, 90 nm and 65 nm CMOS technologies. The NF of IDCLNA with and without parasitic overlap capacitances are analyzed and compared for different short channel CMOS processes. Simulation results show that the parasitic overlap capacitances have advantageous to reduce the gate induced noise in IDCLNA for 130-nm CMOS process for 2.4 GHz applications.
Agglomeration-based physical frame dG discretizations: An attempt to be mesh free
Authors: Bassi Francesco , Botti Lorenzo , Colombo Alessandro
Published: January 2014

In this work we consider agglomeration-based physical frame discontinuous Galerkin (dG) discretization as an effective way to increase the flexibility of high-order finite element methods. The mesh free concept is pursued in the following (broad) sense: the computational domain is still discretized using a mesh but the computational grid should not be a constraint for the finite element discretization. In particular the discrete space choice, its convergence properties, and even the complexity of solving the global system of equations resulting from the dG discretization should not be influenced by the grid choice. Physical frame dG discretization allows to obtain mesh-independent h-convergence rates. Thanks to mesh agglomeration, high-order accurate discretizations can be performed on arbitrarily coarse grids, without resorting to very high-order approximations of domain boundaries. Agglomeration-based h-multigrid techniques are the obvious choice to obtain fast and grid-independent solvers. These features (attractive for any mesh free discretization) are demonstrated in practice with numerical test cases.
New perspectives on polygonal and polyhedral finite element methods
Authors: Manzini Gianmarco , Russo Alessandro , Sukumar N.
Published: January 2014

Generalized barycentric coordinates such as Wachspress and mean value coordinates have been used in polygonal and polyhedral finite element methods. Recently, mimetic finite difference schemes were cast within a variational framework, and a consistent and stable finite element method on arbitrary polygonal meshes was devised. The method was coined as the virtual element method (VEM), since it did not require the explicit construction of basis functions. This advance provides a more in-depth understanding of mimetic schemes, and also endows polygonal-based Galerkin methods with greater flexibility than three-node and four-node finite element methods. In the VEM, a projection operator is used to realize the decomposition of the stiffness matrix into two terms: a consistent matrix that is known, and a stability matrix that must be positive semi-definite and which is only required to scale like the consistent matrix. In this paper, we first present an overview of previous developments on conforming polygonal and polyhedral finite elements, and then appeal to the exact decomposition in the VEM to obtain a robust and efficient generalized barycentric coordinate-based Galerkin method on polygonal and polyhedral elements. The consistent matrix of the VEM is adopted, and numerical quadrature with generalized barycentric coordinates is used to compute the stability matrix. This facilitates post-processing of field variables and visualization in the VEM, and on the other hand, provides a means to exactly satisfy the patch test with efficient numerical integration in polygonal and polyhedral finite elements. We present numerical examples that demonstrate the sound accuracy and performance of the proposed method. For Poisson problems in ℝ2 and ℝ3, we establish that linearly complete generalized barycentric interpolants deliver optimal rates of convergence in the L2-norm and the H1-seminorm.
BLACK HOLE THERMODYNAMIC FLUCTUATIONS AND PHASE TRANSITION OF IR MODIFIED HOŘAVA–LIFSHITZ GRAVITY
Authors: ZHOU SHIWEI , NIU ZHENFENG , LÜ YAN
Published: January 2014

Hořava–Lifshitz theory as a renormalizable model of gravity might be an ultraviolet (UV) completion of general relativity (GR) and it reduces to Einstein gravity with a nonvanishing cosmological constant in infrared (IR) approximation. Kehagias and Sfetsos have added a relevant operator proportional to the three-dimensional (3D) geometry Ricci scalar to the original Hořava–Lifshitz theory action and obtained a spherically symmetric asymptotically flat black hole solution called Kehagias–Sfetsos (KS) black hole. Nonequilibrium thermodynamic fluctuations based on the metric of a KS black hole in IR modified Hořava–Lifshitz gravity have been calculated. It is concluded that the second-order momentum of mass flux is nondivergent and phase transition does not occur at the extremal case, while phase transition occurs at some other case, which is also different from the common case when the heat capacity is divergent.
HAWKING RADIATION OF DIRAC PARTICLES FROM A GENERAL DYNAMICAL SPHERICALLY SYMMETRICAL BLACK HOLE USING A NEW TORTOISE COORDINATE TRANSFORMATION
Authors: LIANG JUN , ZHANG FANG-HUI , ZHANG WEI , ZHANG JING
Published: January 2014

By utilizing the improved Damour–Ruffini method with a new tortoise transformation, we study the Hawking radiation of Dirac particles from a general dynamical spherically symmetric black hole. In the improved Damour–Ruffini method, the position of the event horizon of the black hole is an undetermined function, and the temperature parameter κ is an undetermined constant. By requiring the Dirac equation to be the standard wave equation near the event horizon of the black hole, κ can be determined automatically. Therefore, the Hawking temperature can be obtained. The result is consistent with that of the Hawking radiation of scalar particles.
EFFECT OF KOH ON MICRO-ARC OXIDATION COATINGS OF 2A12 ALUMINUM ALLOYS IN CH3COONa–Na2WO4 ELECTROLYTE
Authors: LIN ZHAO QING , YU HUI JUN , HE SI YU , HE YI NING , CHEN CHUAN ZHONG
Published: January 2014

The ceramic coatings were prepared on 2A12 alloy by micro-arc oxidation in CH 3 COONa – Na 2 WO 4 electrolyte system with different concentration of KOH added. The effects of KOH in this electrolyte on micromorphology, phase compositions, adhesion and corrosion resistance of the coatings were investigated by scanning electron microscope (SEM), X-ray diffraction (XRD), scratch test and electrochemistry workstation. The results show that KOH has a significant influence on the surface morphology, which can make the surface smoother. The adhesion of the coating becomes stronger with the increase of KOH in the electrolyte. The corrosion resistance of the coated specimen increases a lot compared with that of the substrate. And the lowest corrosion current density (I corr ) of the coating prepared in the electrolyte with KOH is about three orders of magnitude lower than that of the substrate.
POLYMERIC 1 × 2 DIGITAL OPTICAL AND 2 × 2 MACH–ZEHNDER INTERFEROMETER WAVEGUIDE THERMO-OPTIC SWITCHES: DESIGN AND SIMULATION
Authors: WU GUANGYU , WANG HAIBO
Published: January 2014

In this paper, a polyurethane (PU) was prepared from polyether polyols, isophorone diisocyanate and 2-[4-( N , N -hydroxyethyl)-aminophenyl-azo]-6-nitrobenzothiazole. The Tg and Td (5% mass loss) are 112°C and 273°C, respectively. The refractive index, transmission loss and Sellmyer coefficients of PU were investigated. Polymeric 1 × 2 digital optical switch (DOS) and 2 × 2 Mach–Zehnder interferometer (MZI) thermo-optic switches based on PU as waveguide at the infrared communication wavelength 1.55 μm were designed and simulated. Simulation results showed that the power consumption of 1 × 2 DOS is only 0.60 mW; and the response times of 1 × 2 and MZI switches are 9 ms and 2.5 ms, respectively.
OPTICAL, THERMAL AND MECHANICAL STUDIES ON NONLINEAR OPTICAL MATERIAL DIGLYCINE BARIUM CHLORIDE MONOHYDRATE (DGBCM) SINGLE CRYSTAL
Authors: MARUDHU G. , KRISHNAN S. , THILAK T. , SAMUEL P. , VINITHA G. , PASUPATHI G.
Published: January 2014

The nonlinear optical single crystals of diglycine barium chloride monohydrate were grown by slow evaporation solution growth technique from the mixture of an aqueous solution of glycine and barium chloride in the ratio 2:1 at room temperature. The grown crystals were characterized by various techniques such as single crystals X-ray diffraction, FTIR, UV-Vis-NIR spectra, Vickers hardness studies, thermogravimetric analysis, and fluorescence spectra. Its second harmonic generation efficiency was measured by Kurtz and Perry powder technique using Nd :YAG laser.