bluuebunny/crossref_metadata_2025 huggingface dataset:
Evaluation of Quantitative Antibiotic Susceptibility Testing by Vitek 2 as a Routine Method To Predict Strain Relatedness of Coagulase-Negative Staphylococci Isolated from Blood Cultures
Authors: Camerer Antonia , Kohlenberg Anke , Stefanik Danuta , Seifert Harald
Published: July 2011

To test the hypothesis that the strain relatedness of coagulase-negative staphylococci (CoNS) recovered from blood cultures can be inferred from automated antibiotic susceptibility testing (AST) results generated by Vitek 2, concordant or discordant AST results were compared with pulsed-field gel electrophoresis (PFGE) typing results for 119 CoNS blood culture isolate pairs. Concordant AST results were highly predictive of the strain relatedness of CoNS isolates.
Deletion of Kaposi's Sarcoma-Associated Herpesvirus FLICE Inhibitory Protein, vFLIP, from the Viral Genome Compromises the Activation of STAT1-Responsive Cellular Genes and Spindle Cell Formation in Endothelial Cells
Authors: Alkharsah Khaled R. , Singh Vivek Vikram , Bosco Raffaella , Santag Susann , Grundhoff Adam , Konrad Andreas , Stürzl Michael , Wirth Dagmar , Dittrich-Breiholz Oliver , Kracht Michael , Schulz Thomas F.
Published: July 2011

Kaposi's sarcoma herpesvirus (KSHV) Fas-associated death domain (FADD)-like interleukin-1 beta-converting enzyme (FLICE)-inhibitory protein, vFLIP, has antiapoptotic properties, is a potent activator of the NF-κB pathway, and induces the formation of endothelial spindle cells, the hallmark of Kaposi's sarcoma, when overexpressed in primary endothelial cells. We used a reverse genetics approach to study several functions of KSHV vFLIP in the context of the whole viral genome. Deletion of the gene encoding vFLIP from a KSHV genome cloned in a bacterial artificial chromosome (BAC) reduced the ability of the virus to persist and induce spindle cell formation in primary human umbilical vein endothelial cells (HUVECs). Only a few, mainly interferon (IFN)-responsive, genes were expressed in wild-type KSHV (KSHV-wt)-infected endothelial cells at levels higher than those in KSHV-ΔFLIP-infected endothelial cells, in contrast to the plethora of cellular genes induced by overexpressed vFLIP. In keeping with this observation, vFLIP induces the phosphorylation of STAT1 and STAT2 in an NF-κB-dependent manner in endothelial cells. vFLIP-dependent phosphorylation of STAT1 and STAT2 could be demonstrated after endothelial cells were infected with KSHV-wt, KSHV-ΔFLIP, and a KSHV-vFLIP revertant virus. These findings document the impact of KSHV vFLIP on the transcriptome of primary endothelial cells during viral persistence and highlight the role of vFLIP in the activation of STAT1/STAT2 and STAT-responsive cellular genes by KSHV.
Mechanisms Controlling Virulence Thresholds of Mixed Viral Populations
Authors: Lancaster Karen Z. , Pfeiffer Julie K.
Published: July 2011

The propensity of RNA viruses to revert attenuating mutations contributes to disease and complicates vaccine development. Despite the presence of virulent revertant viruses in some live-attenuated vaccines, disease from vaccination is rare. This suggests that in mixed viral populations, attenuated viruses may limit the pathogenesis of virulent viruses, thus establishing a virulence threshold. Here we examined virulence thresholds using mixtures of virulent and attenuated viruses in a transgenic mouse model of poliovirus infection. We determined that a 1,000-fold excess of the attenuated Sabin strain of poliovirus was protective against disease induced by the virulent Mahoney strain. Protection was induced locally, and inactivated virus conferred protection. Treatment with a poliovirus receptor-blocking antibody phenocopied the protective effect of inactivated viruses
in vitro
and
in vivo
, suggesting that one mechanism controlling virulence thresholds may be competition for a viral receptor. Additionally, the type I interferon response reduces poliovirus pathogenesis; therefore, we examined virulence thresholds in mice lacking the alpha/beta interferon receptor. We found that the attenuated virus was virulent in immunodeficient mice due to the enhanced replication and reversion of attenuating mutations. Therefore, while the type I interferon response limits the virulence of the attenuated strain by reducing replication, protection from disease conferred by the attenuated strain in immunocompetent mice can occur independently of replication. Our results identified mechanisms controlling the virulence of mixed viral populations and indicate that live-attenuated vaccines containing virulent virus may be safe, as long as virulent viruses are present at levels below a critical threshold.
Emergence of Avian Influenza Viruses with Enhanced Transcription Activity by a Single Amino Acid Substitution in the Nucleoprotein during Replication in Chicken Brains
Authors: Tada Tatsuya , Suzuki Koutaro , Sakurai Yu , Kubo Masanori , Okada Hironao , Itoh Toshihiro , Tsukamoto Kenji
Published: July 2011

To explore the genetic basis of the pathogenesis and adaptation of avian influenza viruses (AIVs) to chickens, the A/duck/Yokohama/aq10/2003 (H5N1) (DkYK10) virus was passaged five times in the brains of chickens. The brain-passaged DkYK10-B5 caused quick death of chickens through rapid and efficient replication in tissues, accompanied by severe apoptosis. Genome sequence comparison of two viruses identified a single amino acid substitution at position 109 in NP from isoleucine to threonine (NP
I
109
T
). By analyzing viruses constructed by the reverse-genetic method, we established that the NP
I
109
T
substitution also contributed to increased viral replication and polymerase activity in chicken embryo fibroblasts, but not in duck embryo fibroblasts. Real-time RT-PCR analysis demonstrated that the NP
I
109
T
substitution enhances mRNA synthesis quickly and then cRNA and viral RNA (vRNA) synthesis slowly. Next, to determine the mechanism underlying the appearance of the NP
I
109
T
substitution during passages, four H5N1 highly pathogenic AIVs (HPAIVs) were passaged in the lungs and brains of chicken embryos. Single-nucleotide polymorphism analysis, together with a database search, suggests that the NP
I
109
T
mutation would be induced frequently during replication of HPAIVs in brains, but not in lungs. These results demonstrate that the amino acid at position 109 in NP enhances viral RNA synthesis and the pathogenicity of highly pathogenic avian influenza viruses in chickens and that the NP mutation emerges quickly during replication of the viruses in chicken brains.
Widespread Endogenization of Densoviruses and Parvoviruses in Animal and Human Genomes
Authors: Liu Huiquan , Fu Yanping , Xie Jiatao , Cheng Jiasen , Ghabrial Said A. , Li Guoqing , Peng Youliang , Yi Xianhong , Jiang Daohong
Published: July 2011

Parvoviruses infect humans and a broad range of animals, from mammals to crustaceans, and generally are associated with a variety of acute and chronic diseases. However, many others cause persistent infections and are not known to be associated with any disease. Viral persistence is likely related to the ability to integrate into the chromosomal DNA and to establish a latent infection. However, there is little evidence for genome integration of parvoviral DNA except for
Adeno-associated virus
(AAV). Here we performed a systematic search for homologs of parvoviral proteins in publicly available eukaryotic genome databases followed by experimental verification and phylogenetic analysis. We conclude that parvoviruses have frequently invaded the germ lines of diverse animal species, including mammals, fishes, birds, tunicates, arthropods, and flatworms. The identification of orthologous endogenous parvovirus sequences in the genomes of humans and other mammals suggests that parvoviruses have coexisted with mammals for at least 98 million years. Furthermore, some of the endogenized parvoviral genes were expressed in eukaryotic organisms, suggesting that these viral genes are also functional in the host genomes. Our findings may provide novel insights into parvovirus biology, host interactions, and evolution.
The Major Determinant of Attenuation in Mice of the Candid1 Vaccine for Argentine Hemorrhagic Fever Is Located in the G2 Glycoprotein Transmembrane Domain
Authors: Albariño César G. , Bird Brian H. , Chakrabarti Ayan K. , Dodd Kimberly A. , Flint Mike , Bergeron Éric , White David M. , Nichol Stuart T.
Published: July 2011

Candid1, a live-attenuated Junin virus vaccine strain, was developed during the early 1980s to control Argentine hemorrhagic fever, a severe and frequently fatal human disease. Six amino acid substitutions were found to be unique to this vaccine strain, and their role in virulence attenuation in mice was analyzed using a series of recombinant viruses. Our results indicate that Candid1 is attenuated in mice through a single amino acid substitution in the transmembrane domain of the G2 glycoprotein. This work provides insight into the molecular mechanisms of attenuation of the only arenavirus vaccine currently available.
Expression of Herpes Simplex Virus 1-Encoded MicroRNAs in Human Trigeminal Ganglia and Their Relation to Local T-Cell Infiltrates
Authors: Held Kathrin , Junker Andreas , Dornmair Klaus , Meinl Edgar , Sinicina Inga , Brandt Thomas , Theil Diethilde , Derfuss Tobias
Published: July 2011

Herpes simplex type 1 (HSV-1) is a neurotropic virus which establishes lifelong latency in human trigeminal ganglia (TG). Currently, two nonexclusive control mechanisms of HSV-1 latency are discussed: antiviral CD8
+
T cells and viral microRNAs (miRNAs) encoded by the latency associated transcript (LAT). We investigate here to what extent these mechanisms may contribute to the maintenance of HSV-1 latency. We show that only a small proportion of LAT
+
neurons is surrounded by T cells in human TG. This indicates that viral latency in human TG might be controlled by other mechanisms such as viral miRNAs. Therefore, we assessed TG sections for the presence of HSV-1 miRNA, DNA, and mRNA by combining LAT
in situ
hybridization, T-cell immunohistochemistry, and single cell analysis of laser-microdissected sensory neurons. Quantitative reverse transcription-PCR (RT-PCR) revealed that LAT
+
neurons with or without surrounding T cells were always positive for HSV-1 miRNAs and DNA. Furthermore, ICP0 mRNA could rarely be detected only in LAT
+
neurons, as analyzed by single-cell RT-PCR. In contrast, in LAT
−
neurons that were surrounded by T cells, neither miRNAs nor the DNA of HSV-1, HSV-2, or varicella-zoster virus could be detected. These data indicate that the majority of LAT
+
neurons is not directly controlled by T cells. However, miRNA expression in every latently infected neuron would provide an additional checkpoint before viral replication is initiated.
The Fecal Viral Flora of California Sea Lions
Authors: Li Linlin , Shan Tongling , Wang Chunlin , Côté Colette , Kolman John , Onions David , Gulland Frances M. D. , Delwart Eric
Published: July 2011

California sea lions are one of the major marine mammal species along the Pacific coast of North America. Sea lions are susceptible to a wide variety of viruses, some of which can be transmitted to or from terrestrial mammals. Using an unbiased viral metagenomic approach, we surveyed the fecal virome in California sea lions of different ages and health statuses. Averages of 1.6 and 2.5 distinct mammalian viral species were shed by pups and juvenile sea lions, respectively. Previously undescribed mammalian viruses from four RNA virus families (Astroviridae,Picornaviridae,Caliciviridae, andReoviridae) and one DNA virus family (Parvoviridae) were characterized. The first complete or partial genomes of sapeloviruses, sapoviruses, noroviruses, and bocavirus in marine mammals are reported. Astroviruses and bocaviruses showed the highest prevalence and abundance in California sea lion feces. The diversity of bacteriophages was higher in unweaned sea lion pups than in juveniles and animals in rehabilitation, where the phage community consisted largely of phages related to the familyMicroviridae. This study increases our understanding of the viral diversity in marine mammals, highlights the high rate of enteric viral infections in these highly social carnivores, and may be used as a baseline viral survey for comparison with samples from California sea lions during unexplained disease outbreaks.
Epstein-Barr Virus Latent Membrane Protein 1 (LMP1) C-Terminal-Activating Region 3 Contributes to LMP1-Mediated Cellular Migration via Its Interaction with Ubc9
Authors: Bentz Gretchen L. , Whitehurst Christopher B. , Pagano Joseph S.
Published: July 2011

Epstein-Barr virus (EBV) latent membrane protein 1 (LMP1), the principal viral oncoprotein and a member of the tumor necrosis factor receptor superfamily, is a constitutively active membrane signaling protein that regulates multiple signal transduction pathways via its C-terminal-activating region 1 (CTAR1) and CTAR2, and also the less-studied CTAR3. Because protein sumoylation among other posttranslational modifications may regulate many signaling pathways induced by LMP1, we investigated whether during EBV latency LMP1 regulates sumoylation processes that control cellular activation and cellular responses. By immunoprecipitation experiments, we show that LMP1 interacts with Ubc9, the single reported SUMO-conjugating enzyme. Requirements for LMP1-Ubc9 interactions include enzymatically active Ubc9: expression of inactive Ubc9 (Ubc9 C93S) inhibited the LMP1-Ubc9 interaction. LMP1 CTAR3, but not CTAR1 and CTAR2, participated in the LMP1-Ubc9 interaction, and amino acid sequences found in CTAR3, including the JAK-interacting motif, contributed to this interaction. Furthermore, LMP1 expression coincided with increased sumoylation of cellular proteins, and disruption of the Ubc9-LMP1 CTAR3 interaction almost completely abrogated LMP1-induced protein sumoylation, suggesting that this interaction promotes the sumoylation of downstream targets. Additional consequences of the disruption of the LMP1 CTAR3-Ubc9 interaction revealed effects on cellular migration, a hallmark of oncogenesis. Together, these data demonstrate that LMP1 CTAR3 does in fact function in intracellular signaling and leads to biological effects. We propose that LMP1, by interaction with Ubc9, modulates sumoylation processes, which regulate signal transduction pathways that affect phenotypic changes associated with oncogenesis.
Cross-Clade HIV-1 Neutralizing Antibodies Induced with V3-Scaffold Protein Immunogens following Priming with gp120 DNA
Authors: Zolla-Pazner Susan , Kong X.-P. , Jiang Xunqing , Cardozo Timothy , Nádas Arthur , Cohen Sandra , Totrov Maxim , Seaman Michael S. , Wang Shixia , Lu Shan
Published: July 2011

The V3 epitope is a known target for HIV-1 neutralizing antibodies (NAbs), and V3-scaffold fusion proteins used as boosting immunogens after gp120 DNA priming were previously shown to induce NAbs in rabbits. Here, we evaluated whether the breadth and potency of the NAb response could be improved when boosted with rationally designed V3-scaffold immunogens. Rabbits were primed with codon-optimized clade C gp120 DNA and boosted with one of five V3-cholera toxin B fusion proteins (V3-CTBs) or with double combinations of these. The inserts in these immunogens were designed to display V3 epitopes shared by the majority of global HIV-1 isolates. Double combinations of V3-CTB immunogens generally induced more broad and potent NAbs than did boosts with single V3-CTB immunogens, with the most potent and broad NAbs elicited with the V3-CTB carrying the consensus V3 of clade C (V3
C
-CTB), or with double combinations of V3-CTB immunogens that included V3
C
-CTB. Neutralization of tier 1 and 2 pseudoviruses from clades AG, B, and C and of peripheral blood mononuclear cell (PBMC)-grown primary viruses from clades A, AG, and B was achieved, demonstrating that priming with gp120 DNA followed by boosts with V3-scaffold immunogens effectively elicits cross-clade NAbs. Focusing on the V3 region is a first step in designing a vaccine targeting protective epitopes, a strategy with potential advantages over the use of Env, a molecule that evolved to protect the virus by poorly inducing NAbs and by shielding the epitopes that are most critical for infectivity.
Identification of Nuclear and Nucleolar Localization Signals of Pseudorabies Virus (PRV) Early Protein UL54 Reveals that Its Nuclear Targeting Is Required for Efficient Production of PRV
Authors: Li Meili , Wang Shuai , Cai Mingsheng , Zheng Chunfu
Published: July 2011

The pseudorabies virus (PRV) early protein UL54 is a homologue of herpes simplex virus 1 (HSV-1) immediate-early protein ICP27, which is a multifunctional protein that is essential for HSV-1 infection. In this study, the subcellular localization and nuclear import signals of PRV UL54 were characterized. UL54 was shown to predominantly localize to the nucleolus in transfected cells. By constructing a series of mutants, a functional nuclear localization signal (NLS) and a genuine nucleolar localization signal (NoLS) of UL54 were for the first time identified and mapped to amino acids
61
RQRRR
65
and
45
RRRRGGRGGRAAR
57
, respectively. Additionally, three recombinant viruses with mutations of the NLS and/or the NoLS in UL54 were constructed based on PRV bacterial artificial chromosome (BAC) pBecker2 to test the effect of UL54 nuclear targeting on viral replication. In comparison with the wild-type virus, a recombinant virus harboring an NLS or NoLS mutation of UL54 reduced viral production to different extents. However, mutations of both the NLS and NoLS targeted UL54 to the cytoplasm in recombinant virus-infected cells and significantly impaired viral replication, comparable to the UL54-null virus. In addition, a virus lacking the NLS or the NoLS displayed modest defects in viral gene expression and DNA synthesis. However, deletion of both the NLS and the NoLS resulted in severe defects in viral gene expression and DNA synthesis, as well as production of infectious progeny. Thus, we have identified a classical NLS and a genuine NoLS in UL54 and demonstrate that the nuclear targeting of UL54 is required for efficient production of PRV.
Creation of a Recombinant Rift Valley Fever Virus with a Two-Segmented Genome
Authors: Brennan Benjamin , Welch Stephen R. , McLees Angela , Elliott Richard M.
Published: July 2011

Rift Valley fever virus (RVFV; family
Bunyaviridae
) is a clinically important, mosquito-borne pathogen of both livestock and humans, which is found mainly in sub-Saharan Africa and the Arabian Peninsula. RVFV has a trisegmented single-stranded RNA (ssRNA) genome. The L and M segments are negative sense and encode the L protein (viral polymerase) on the L segment and the virion glycoproteins Gn and Gc as well as two other proteins, NSm and 78K, on the M segment. The S segment uses an ambisense coding strategy to express the nucleocapsid protein, N, and the nonstructural protein, NSs. Both the NSs and NSm proteins are dispensable for virus growth in tissue culture. Using reverse genetics, we generated a recombinant virus, designated r2segMP12, containing a two-segmented genome in which the NSs coding sequence was replaced with that for the Gn and Gc precursor. Thus, r2segMP12 lacks an M segment, and although it was attenuated in comparison to the three-segmented parental virus in both mammalian and insect cell cultures, it was genetically stable over multiple passages. We further show that the virus can stably maintain an M-like RNA segment encoding the enhanced green fluorescent protein gene. The implications of these findings for RVFV genome packaging and the potential to develop multivalent live-attenuated vaccines are discussed.
Reassortment Events among Swine Influenza A Viruses in China: Implications for the Origin of the 2009 Influenza Pandemic
Authors: Lam Tommy Tsan-Yuk , Zhu Huachen , Wang Jia , Smith David K. , Holmes Edward C. , Webster Robert G. , Webby Richard , Peiris Joseph M. , Guan Yi
Published: July 2011

That pigs may play a pivotal role in the emergence of pandemic influenza was indicated by the recent H1N1/2009 human pandemic, likely caused by a reassortant between viruses of the American triple-reassortant (TR) and Eurasian avian-like (EA) swine influenza lineages. As China has the largest human and pig populations in the world and is the only place where both TR and EA viruses have been reported to cocirculate, it is potentially the source of the H1N1/2009 pandemic virus. To examine this, the genome sequences of 405 swine influenza viruses from China were analyzed. Thirty-six TR and EA reassortant viruses were identified before and after the occurrence of the pandemic. Several of these TR-EA reassortant viruses had genotypes with most segments having the same lineage origin as the segments of the H1N1/2009 pandemic virus. However, these viruses were generated from independent reassortment events throughout our survey period and were not associated with the current pandemic. One TR-EA reassortant, which is least similar to the pandemic virus, has persisted since 2007, while all the other variants appear to be transient. Despite frequent reassortment events between TR and EA lineage viruses in China, evidence for the genesis of the 2009 pandemic virus in pigs in this region is still absent.
Circulating Monocytes Are Not a Major Reservoir of HIV-1 in Elite Suppressors
Authors: Spivak Adam M. , Salgado Maria , Rabi S. Alireza , O'Connell Karen A. , Blankson Joel N.
Published: July 2011

Circulating HIV-1-infected monocytes have been identified in patients on highly active antiretroviral therapy and may represent an important barrier to viral eradication. The nature of these cells in HIV-1-infected patients who maintain undetectable viral loads and preserved CD4
+
T cell counts without antiretroviral therapy (known as elite controllers or elite suppressors [ES]) is unknown. We describe here infrequent recovery of proviral HIV-1 DNA from circulating monocytes relative to CD4
+
T cells in ES, despite permissiveness of these cells to HIV-1 viral entry
ex vivo
. Thus, monocytes do not appear to be a major reservoir of HIV-1 in ES.
Herpesvirus Entry Mediator and Nectin-1 Mediate Herpes Simplex Virus 1 Infection of the Murine Cornea
Authors: Karaba Andrew H. , Kopp Sarah J. , Longnecker Richard
Published: July 2011

Herpes simplex virus 1 (HSV-1) is a ubiquitous human pathogen that enters cells by the receptor-mediated fusion of the viral envelope with a host cell membrane. The envelope glycoprotein gD of HSV must bind to one of its receptors for entry to take place. Recent studies using knockout (KO) mice demonstrated that the gD receptors herpesvirus entry mediator (HVEM) and nectin-1 are the primary entry receptors for HSV-2 in the mouse vagina and brain. Nectin-1 was most crucial for the neuronal spread of HSV-2, particularly in the brain. HVEM was dispensable for infection in these models, but when both HVEM and nectin-1 were absent, infection was completely prevented. We sought to determine the receptor requirements of HSV-1 in an ocular model of infection using knockout mice. Wild-type, HVEM KO, nectin-1 KO, and HVEM/nectin-1 double-KO mice were infected via corneal scarification and monitored for clinical signs of infection and viral replication in various tissues. We report that either HVEM or nectin-1 must be present for HSV-1 infection of the cornea. Additionally, we observed that the infection was attenuated in both HVEM KO and nectin-1 KO mice. This is in contrast to what was reported for studies of HSV-2 in vagina and brain and suggests that receptor requirements for HSV vary depending on the route of inoculation and/or serotype.
Longitudinal Analysis of the Human T Cell Response during Acute Hantavirus Infection
Authors: Lindgren Therese , Ahlm Clas , Mohamed Nahla , Evander Magnus , Ljunggren Hans-Gustaf , Björkström Niklas K.
Published: July 2011

Longitudinal studies of T cell immune responses during viral infections in humans are essential for our understanding of how effector T cell responses develop, clear infection, and provide long-lasting immunity. Here, following an outbreak of a Puumala hantavirus infection in the human population, we longitudinally analyzed the primary CD8 T cell response in infected individuals from the first onset of clinical symptoms until viral clearance. A vigorous CD8 T cell response was observed early following the onset of clinical symptoms, determined by the presence of high numbers of Ki67+CD38+HLA-DR+effector CD8 T cells. This response encompassed up to 50% of total blood CD8 T cells, and it subsequently contracted in parallel with a decrease in viral load. Expression levels of perforin and granzyme B were high throughout the initial T cell response and likewise normalized following viral clearance. When monitoring regulatory components, no induction of regulatory CD4 or CD8 T cells was observed in the patients during the infection. However, CD8 as well as CD4 T cells exhibited a distinct expression profile of inhibitory PD-1 and CTLA-4 molecules. The present results provide insight into the development of the T cell response in humans, from the very onset of clinical symptoms following a viral infection to resolution of the disease.
Counting your losses: medicines audits in practice
Authors: Macfarlane John , Robson Rachel
Published: July 2011

It has been estimated that shrinkage of veterinary purchases – that is, the difference between what is bought and sold – is in the order of 5 to 10 per cent. XLVets' key performance indicator data suggest that medicines sales per vet by sector are £81,500 (small animal), £147,000 (farm animal) and £44,500 (equine); 10 or even 5 per cent of these amounts would create a significant loss to the bottom line of any business. The only way of revealing the true extent of these losses in your practice is to conduct a full medicines audit, and this must be adopted as a regular feature of stock management. In this article, John Macfarlane and Rachel Robson describe how to carry out a medicines audit.
Investigation and management of cervical swellings in dogs
Authors: Owen Laura
Published: July 2011

Cervical swellings in the dog comprise neoplastic masses, abscesses/granulomas, salivary mucoceles and congenital cysts. It is important to differentiate between these conditions because the investigative pathway, treatment and prognosis for each of them are very different. Accurate history taking, physical examination and cytology can all provide vital clues about the nature of the disease and, hence, often avoid the need for more invasive and expensive testing. Subsequent decision‐making depends on patient, owner and disease factors as well as financial constraints. This article describes how to diagnose cervical swellings in dogs and highlights the management options for each type of condition.
Problems associated with cattle welfare
Authors: Gregory Neville
Published: July 2011

Recent studies have highlighted six welfare issues affecting cattle: assessing prognosis in cows with uterine prolapse; methods for feeding calves colostrum; understanding the relationship between floor hygiene, slurry heel and mastitis; managing slippery floors; avoiding foot disorders caused by newly laid concrete; and genetic antagonisms between production performance and cow welfare. This article describes how these problems might be prevented and discusses the role of the veterinary surgeon in educating farmers on how to manage issues affecting cattle welfare appropriately.
Speed cameras in an urban setting: a cost–benefit analysis
Authors: Mendivil Joan , García-Altés Anna , Pérez Katherine , Marí-Dell'Olmo Marc , Tobías Aurelio
Published: July 2011

Background

To perform a cost–benefit analysis of the installation of speed cameras on the beltways of Barcelona.

Methods

The analysis was performed from the society perspective over a 2-year period using a controlled before-and-after study design. The net benefit was calculated using, as benefits, the willingness to pay for the estimated number of people who avoided injury as a consequence of the intervention, subtracting costs and savings. Sensitivity analyses were performed using the maximum and minimum estimated number of people who avoided injury, assuming that there was a minimum of one death avoided, using the maximum value of a statistical life, assigning all implementation costs to the first year and assuming there was no time lost due to speed reduction.

Results

Base case results showed a net benefit of €6.8 million. Sensitivity analyses suggested that net benefits could range from €5.6 to €23.1 million over 2 years.

Conclusions

The use of speed cameras in urban areas has a favourable economic impact even when assessed using conservative assumptions.
Unmet need for effective family planning in HIV-infected individuals: results from a survey in rural Uganda
Authors: Jhangri Gian S , Heys Jennifer , Alibhai Arif , Rubaale Tom , Kipp Walter
Published: July 2011

Background and methodology

This study determined the unmet need for family planning among HIV-positive and HIV-negative individuals living in western Uganda. Semi-structured interviews were conducted with individuals who were randomly selected from HIV testing lists. Of those individuals, further analysis was conducted on a subset of 206 participants who did not desire more children and were not using a highly effective method of contraception. Descriptive, bivariate and multivariate methods were performed to assess the relationship between HIV status and unmet need for effective family planning.

Results

The unmet need for effective family planning was much greater in HIV-infected individuals compared to HIV-negative individuals [75.0% vs 33.8%, adjusted odds ratio (OR) 3.97, 95% confidence interval (CI) 1.97–8.03, p<0.001]. Females were more likely to report an unmet need compared to males (69.0% vs 49.5%; adjusted OR 1.94, 95% CI 0.94–4.00, p=0.071). Other predictors of unmet need for effective family planning were older age (adjusted OR 1.08 for each year of age, 95% CI 1.00–1.16, p=0.018) and single/cohabiting vs being married (OR 2.36, 95% CI 1.16–4.80, p=0.036). Being on antiretroviral therapy was not a predictor of having a lower unmet need for effective family planning methods.

Discussion and conclusions

There is high unmet need for effective family planning in HIV-positive study participants in a region of western Uganda, which should be of concern. This suggests that HIV-infected individuals do not want to use family planning or encounter barriers to accessing and utilising family planning services. Family planning programmes and HIV care and prevention services have to work together more effectively to create services conducive to clients from both programmes.
Acceptability of self-testing for trichomoniasis increases with experience
Authors: Huppert Jill S , Hesse Elizabeth A , Bernard Molly A , Xiao Yang , Huang Bin , Gaydos Charlotte A , Kahn Jessica A
Published: July 2011

Objectives

Self-testing and point-of-care (POC) tests could improve the detection of sexually transmitted infections (STIs) in adolescents. This study aimed to (1) validate a scale measuring acceptability of self-testing for trichomoniasis, (2) compare acceptability of self versus clinician testing using a POC test for trichomoniasis, (3) examine changes in acceptability after experience and review of results, and (4) examine predictors of acceptability.

Methods

Women (14–22 years old) performed the POC test and completed surveys assessing acceptability of self and clinician testing at baseline, after testing, and after discussion of results. Factor analysis examined scale structure; changes in mean scale scores were assessed with mixed models. Generalised linear models examined predictors of acceptability.

Results

Of 247 participants, 54 (22%) had a positive POC test for trichomoniasis. Factor analysis confirmed four acceptability subscales: trust of results, confidence, comfort, and effects of testing. At baseline, trust and confidence were higher, and comfort was lower, for clinician versus self testing. For self-testing, all subscale scores increased from baseline to after testing, and trust increased from after testing to after discussion. Trust of self and clinician results was not significantly different after discussion. A positive attitude towards home testing predicted higher self-test acceptability on all subscales. Tampon use was associated with increased self-test comfort at baseline. Concordance between self and clinician results predicted increased trust of self testing after discussion.

Conclusions

Many young women lack confidence in their ability to self-test for trichomoniasis. Allowing women to try a POC test and review the results with a clinician increases acceptability of self-testing.
Feasibility of single‐portal access laparoscopic ovariectomy in 17 cats
Authors: Kim Y‐K. , Lee S‐Y. , Park S‐J. , Lee S‐S. , Lee H‐C. , Lee H‐J. , Yeon S‐C.
Published: July 2011

Laparoscopic ovariectomy (LapOVE) using single‐portal access was attempted in 17 client‐owned cats of different breeds admitted for elective ovariectomy. A 12 mm umbilical portal was placed 1 cm caudal to the umbilicus with the cat in dorsal recumbency. Then, a laparoscope with an operating channel was introduced into the portal with the cat in lateral recumbency. The right ovary was pulled to the abdominal wall using grasping forceps and fixed to the abdominal wall by a transabdominal suspension suture. The ovarian vasculature, suspensory ligament and proper ligament were progressively cauterised and transected with multifunction bipolar electrocoagulation forceps. The resected right ovary was exteriorised through the umbilical portal cannula. The left ovary was then removed from the abdomen in a similar fashion. Surgical time, intraoperative haemorrhage, amount of fat in the ovarian ligament, surgical complications and postoperative pain were recorded. The mean (sd) surgical time was 23 minutes and seven seconds (five minutes and 55 seconds). Intraoperative blood loss and fat deposition of the ovarian ligament were minimal. No intra‐ and postoperative complications were encountered. No cats needed rescue analgesia within 24 hours postsurgery.
Clinicopathological findings and echocardiographic prediction of the localisation of bovine endocarditis
Authors: Mohamed T. , Buczinski S.
Published: July 2011

Thirty‐six cows with confirmed vegetative endocarditis at postmortem examination were examined in this study. In 33 cows, echocardiography showed lesions of the pulmonic, tricuspid and mitral valves. Ten healthy cows were enrolled in the study as controls. In diseased cows, clinical signs included recurrent fever, dyspnoea, epistaxis, tachycardia, murmurs, weight loss, decreased milk production, jugular and mammary vein distension with palpable pulse, and ventral and submandibular oedema. Laboratory abnormalities included neutrophilic leukocytosis, elevated protein concentrations, hypoalbuminaemia, hypergammaglobulinaemia, azotaemia and elevated serum creatinine concentration. High serum activities of aspartate aminotransferase, γ‐glutamyl transpeptidase and lactate dehydrogenase were also detected. Echocardiography showed small, medium‐sized or large hyperechogenicity, thickening, and proliferative or vegetative lesions of the tricuspid (n=11), mitral (n=5) and pulmonic valves (n=19). Confirmatory diagnosis was made at postmortem examination. Echocardiography could detect vegetations in 33 of the 36 cows (92 per cent). The sensitivity of ultrasound in detecting valvular endocarditis was 89 per cent for tricuspid valves, 83 per cent for mitral valves and 95 per cent for pulmonic valves.
CAUSALITY BETWEEN EXPORTS AND PRODUCTIVITY IN THE MALAYSIAN ECONOMY
Authors: IBRAHIM NURHANI ABA
Published: July 2011

Empirical evidence linking exports and productivity growth has been mixed and inconclusive. This study re-examines the direction of the causality between them for Malaysian industries by using the error-correction mechanism and Granger causality models. In a panel of 63 manufacturing industries, for the period of 1981 to 1999, it is found that these industries support the export-led growth and the growth-driven export hypotheses. A further look into the results indicates that there are possibilities of indirect causalities between productivity growth and export through size and capital intensity, as both exports and labor productivity have bidirectional causality with size and capital intensity.
STRUCTURE, EMPLOYMENT AND PRODUCTIVITY GROWTH IN THE INDIAN UNORGANIZED MANUFACTURING SECTOR: AN INDUSTRY LEVEL ANALYSIS
Authors: RAJ S. N. RAJESH
Published: July 2011

The paper analyzes the size, growth and productivity performance of the unorganized manufacturing sector in India during the 1978–1979 to 2000–2001 period. The study shows evidence of an increase in the size of the sector with a slowdown in the reforms period. Evidence indicates that the rate of growth varies widely across the two-digit industries but the variation in growth rate is smaller during the 1990s. Textiles and machinery goods were the fastest growing segments of India's unorganized manufacturing sector in the reforms period. The partial factor productivity approach shows that labor productivity has improved in 2000–2001 over 1978–1979 while capital productivity reported a decline in the same period. The sector, on the other hand, registered a fall in total factor productivity (TFP) during the reforms period. It is found that technological progress has been the main contributor to the growth in TFP in the prereforms period while technical regress contributed to the decline in TFP in the reforms period. A completely different picture is noticed since the mid-1990s when the sector made significant progress in TFP primarily attributed to technological progress which outweighed the decline in technical efficiency. It is also found that capital intensity is an essential factor augmenting labor productivity levels in the sector, which is important for improving the wages paid to the workers in the sector.
FREE TRADE AND GROWTH IN THE WORLD ECONOMY
Authors: LLOYD PETER
Published: July 2011

This paper explores the relationship between free trade and the rate of economic growth. It is argued that freeing trade has both a level effect and a growth effect. Most empirical studies ignore the growth effect and, therefore, considerably understate the beneficial effects of freeing trade. Progress towards free trade in the GATT/WTO era is far from complete. Regionalism has had a limited effect on freeing trade globally. The completion of the Doha Development Round is needed to restart trade as the engine of growth.
Development of the synthesis of the diamine complexes of covalent conjugates of platinum(II) with octacarboxy-substituted cobalt phthalocyanine
Authors: Dolotova Olga , Kaliya Oleg L.
Published: July 2011

Ethylenediamine-N,N′-diacetic acid complexes of the covalent conjugates of octacarboxy-substituted cobalt phthalocyanine with one, two and three platinum atoms (3a–c) were synthesized and characterized by elemental analysis, electronic absorption, infrared and mass spectra data.
CyPPA, a positive modulator of small-conductance Ca2+-activated K+ channels, inhibits phasic uterine contractions and delays preterm birth in mice
Authors: Skarra Dana V. , Cornwell Trudy , Solodushko Viktoriya , Brown Amber , Taylor Mark S.
Published: July 2011

Organized uterine contractions, including those necessary for parturition, are dependent on calcium entry through voltage-gated calcium channels in myometrial smooth muscle cells. Recent evidence suggests that small-conductance Ca2+-activated potassium channels (KCa2), specifically isoforms KCa2.2 and 2.3, may control these contractions through negative feedback regulation of Ca2+ entry. We tested whether selective pharmacologic activation of KCa2.2/2.3 channels might depress uterine contractions, providing a new strategy for preterm labor intervention. Western blot analysis and immunofluorescence microscopy revealed expression of both KCa2.2 and KCa2.3 in the myometrium of nonpregnant (NP) and pregnant (gestation day 10 and 16; D10 and D16, respectively) mice. Spontaneous phasic contractions of isolated NP, D10, and D16 uterine strips were all suppressed by the KCa2.2/2.3-selective activator CyPPA in a concentration-dependent manner. This effect was antagonized by the selective KCa2 inhibitor apamin. Whereas CyPPA sensitivity was reduced in D10 and D16 versus NP strips (pIC50 5.33 ± 0.09, 4.64 ± 0.03, 4.72 ± 0.10, respectively), all contractions were abolished between 30 and 60 μM. Blunted contractions were associated with CyPPA depression of spontaneous Ca2+ events in myometrial smooth muscle bundles. Augmentation of uterine contractions with oxytocin or prostaglandin F2α did not reduce CyPPA sensitivity or efficacy. Finally, in an RU486-induced preterm labor model, CyPPA significantly delayed time to delivery by 3.4 h and caused a 2.5-fold increase in pup retention. These data indicate that pharmacologic stimulation of myometrial KCa2.2/2.3 channels effectively suppresses Ca2+-mediated uterine contractions and delays preterm birth in mice, supporting the potential utility of this approach in tocolytic therapies.
Covalently grafted VEGF165 in hydrogel models upregulates the cellular pathways associated with angiogenesis
Authors: Porter A. M. , Klinge C. M. , Gobin A. S.
Published: July 2011

Angiogenesis is an important biological response known to be involved in many physiological and pathophysiological situations. Cellular responses involved in the formation of new blood vessels, such as increases in endothelial cell proliferation, cell migration, and the survival of apoptosis-inducing events, have been associated with vascular endothelial growth factor isoform 165 (VEGF165). Current research in the areas of bioengineering and biomedical science has focused on developing polyethylene glycol (PEG)-based systems capable of initiating and sustaining angiogenesis in vitro. However, a thorough understanding of how endothelial cells respond at the molecular level to VEGF165 incorporated into these systems has not yet been established in the literature. The goal of the current study was to compare the upregulation of key intracellular proteins involved in angiogenesis in human umbilical vein endothelial cells (HUVEC) and human microvascular endothelial cells (HMEC) seeded on PEG hydrogels containing grafted VEGF165 and adhesion peptides Arg-Gly-Asp-Ser (RGDS). Our data suggest that the covalent incorporation of VEGF165 into PEG hydrogels encourages the upregulation of signaling proteins responsible for increases in endothelial cell proliferation, cell migration, and the survival after apoptosis-inducing events.
KCNQ5/Kv7.5 potassium channel expression and subcellular localization in primate retinal pigment epithelium and neural retina
Authors: Zhang Xiaoming , Yang Dongli , Hughes Bret A.
Published: July 2011

Previous studies identified in retinal pigment epithelial (RPE) cells an M-type K+ current, which in many other cell types is mediated by channels encoded by KCNQ genes. The aim of this study was to assess the expression of KCNQ genes in the monkey RPE and neural retina. Application of the specific KCNQ channel blocker XE991 eliminated the M-type current in freshly isolated monkey RPE cells, indicating that KCNQ subunits contribute to the underlying channels. RT-PCR analysis revealed the expression of KCNQ1, KCNQ4, and KCNQ5 transcripts in the RPE and all five KCNQ transcripts in the neural retina. At the protein level, KCNQ5 was detected in the RPE, whereas both KCNQ4 and KCNQ5 were found in neural retina. In situ hybridization in frozen monkey retinal sections revealed KCNQ5 gene expression in the ganglion cell layer and the inner and outer nuclear layers of the neural retina, but results in the RPE were inconclusive due to the presence of melanin. Immunohistochemistry revealed KCNQ5 in the inner and outer plexiform layers, in cone and rod photoreceptor inner segments, and near the basal membrane of the RPE. The data suggest that KCNQ5 channels contribute to the RPE basal membrane K+ conductance and, thus, likely play an important role in active K+ absorption. The distribution of KCNQ5 in neural retina suggests that these channels may function in the shaping of the photoresponses of cone and rod photoreceptors and the processing of visual information by retinal neurons.
Characterization of FGF receptor expression in human neutrophils and their contribution to chemotaxis
Authors: Haddad Lydia E. , Khzam Lara Bou , Hajjar Fadi , Merhi Yahye , Sirois Martin G.
Published: July 2011

Several members of the fibroblast growth factor (FGF) family are potent endothelial cell (EC) mitogens and angiogenic factors, and their activities can be mediated by four tyrosine kinase receptors (FGFR1–4). In addition, FGFs can induce the release of inflammatory mediators by ECs and the expression of adhesion molecules at their surface, thereby favoring the recruitment and transvascular migration of inflammatory cells such as neutrophils. Neither the expression nor the biological activities that could be mediated by FGFRs have been investigated in human neutrophils. By biochemical and cytological analyses, we observed that purified circulating human neutrophils from healthy individuals expressed varying levels of FGFRs in their cytosol and at their cytoplasmic membrane. FGFR-2 was identified as the sole cell surface receptor, with FGFR-1 and -4 localizing in the cytosol and FGFR-3 being undetectable. We assessed the capacity of FGF-1 and FGF-2 to induce neutrophil chemotaxis in a modified Boyden microchamber and observed that they increase neutrophil transmigration at 10−10 and 10−9 M and by 1.77- and 2.34-fold, respectively, as compared with PBS-treated cells. Treatment with a selective anti-FGFR-2 antibody reduced FGF-1-mediated chemotaxis by 75% and abrogated the effect of FGF-2, while the blockade of FGFR-1 and -4 partially inhibited (15–40%) FGF-chemotactic activities. In summary, our data are the first to report the expression of FGF receptors in human neutrophils, with FGF-1 and FGF-2 promoting neutrophil chemotaxis mainly through FGFR-2 activation.
Persistent detrusor overactivity in rats after relief of partial urethral obstruction
Authors: Jin Long-Hu , Andersson Karl-Erik , Han Jeong-Uk , Kwon Yong-Hyun , Park Chang-Shin , Shin Hwa-Yoan , Yoon Sang-Min , Lee Tack
Published: July 2011

Detrusor overactivity (DO) persists after prostatectomy in 20% to 25% of patients with benign disease. Assuming that nonvoiding contractions (NVCs) can be used as a surrogate for DO in humans, the rat model of obstruction/deobstruction may allow us to study the pathophysiology of persistent DO after deobstruction. We investigated bladder function, with a special focus on NVCs, in rats by use of a new, modified method of obstruction and deobstruction and compared these results with those obtained by use of the conventional method. Seventy female Sprague-Dawley rats underwent 1) sham operation ( n = 10), 2) obstruction by a modified method (Modif-Obs; n = 12), 3) obstruction/deobstruction by the conventional method (Conv-Obs/Deobs; n = 13), or 4) obstruction/deobstruction by the modified method (Modif-Obs/Deobs; n = 35). The Modif-Obs/Deobs animals were divided into subgroups with (DO+) and without (DO−) NVCs. Two weeks after partial urethral obstruction, the animals were deobstructed, and 1 wk later cystometry was performed with recording of intravesical and intra-abdominal pressures. NVCs were shown in all groups: Modif-Obs (80%), Conv-Obs/Deobs (100%), and Modif-Obs/Deobs (40%). In the Modif-Obs/Deobs group, bladder weight and the muscle-to-collagen ratio were higher in DO+ than in DO− rats. The Modif-Obs/Deobs group showed no mortality compared with 25% mortality in the Conv-Obs/Deobs group. The modified method may be more adequate for studying persistent DO after deobstruction, because it resulted in pressure/volume- and DO-related parameters similar to those found in the clinical situation. The persistence of DO after deobstruction may partly be due to irreversible changes in the bladder caused during the period of obstruction.