bluuebunny/crossref_metadata_2025 huggingface dataset:
Generation and Characterization of Chimeric Antibodies against NS3, NS4, NS5, and Core Antigens of Hepatitis C Virus
Authors: Tu Bailin , Ziemann Robert N. , Tieman Bryan C. , Hawksworth David J. , Tyner Joan , Scheffel James , Pinkus Mary S. , Brophy Susan E. , Werneke Jeffrey M. , Gutierrez Robin , White Michael
Published: April 2010

Mouse-human chimeric antibodies (cAbs) against hepatitis C virus (HCV) core, NS3 (nonstructural), NS4, and NS5 antigens were developed as quality control (QC) reagents to replace the use of human sera/plasma for Abbott HCV immunoassays. The cAb retains the mouse monoclonal antibody (MAb) specificity and affinity but still reacts in the existing HCV assay format, which measures human anti-HCV immunoglobulin. Mouse heavy-chain (V
H
) and light-chain (V
L
) variable regions of anti-HCV core, NS3, NS4, and NS5 antigens were PCR amplified from hybridoma lines and then cloned with human IgG1 heavy-chain (C
H
) and light-chain (C
L
) constant regions, respectively. A single mammalian expression plasmid containing both heavy-chain and light-chain immunoglobulin genes was constructed and transfected into dihydrofolate reductase (DHFR)-deficient Chinese hamster ovary (CHO) cells. The transfected CHO cells were selected using hypoxanthine- and thymidine-free medium and screened by an enzyme immunoassay (EIA). The clone secreting the highest level of antibody was isolated from the CHO transfectants and further subcloned. Each cAb-expressing CHO cell line was weaned into serum-free medium, and the cAb was purified by protein A affinity chromatography. The levels of cAb production for the various CHO cell lines varied from 10 to 20 mg/liter. Purified anti-HCV cAbs were tested with Abbott HCV immunoassays and showed reactivity. Moreover, yeast surface display combined with alanine-scanning mutagenesis was used to map the epitope at the individual amino acid level. Our results suggest that these HCV cAbs are ideal controls, calibrators, and/or QC reagents for HCV assay standardization.
Implantable Cardiac Defibrillator Pocket Infection Due to a Previously Undescribed
Cupriavidus
Species
Authors: Christensen Joshua B. , Vitko Nicholas P. , Voskuil Martin I. , Castillo-Mancilla Jose R.
Published: April 2010

The genus
Cupriavidus
consists of Gram-negative, nonfermenting bacteria most of which are environmental organisms, though some species have been associated with human disease. We report the recovery and identification of an isolate that represents a previously undescribed species of
Cupriavidus
from an implantable cardiac defibrillator pocket infection.
Role of Interferon Antagonist Activity of Rabies Virus Phosphoprotein in Viral Pathogenicity
Authors: Ito Naoto , Moseley Gregory W. , Blondel Danielle , Shimizu Kenta , Rowe Caitlin L. , Ito Yuki , Masatani Tatsunori , Nakagawa Keisuke , Jans David A. , Sugiyama Makoto
Published: April 2010

The fixed rabies virus (RV) strain Nishigahara kills adult mice after intracerebral inoculation, whereas the chicken embryo fibroblast cell-adapted strain Ni-CE causes nonlethal infection in adult mice. We previously reported that the chimeric CE(NiP) strain, which has the phosphoprotein (P protein) gene from the Nishigahara strain in the genetic background of the Ni-CE strain, causes lethal infection in adult mice, indicating that the P gene is responsible for the different pathogenicities of the Nishigahara and Ni-CE strains. Previous studies demonstrated that RV P protein binds to the interferon (IFN)-activated transcription factor STAT1 and blocks IFN signaling by preventing its translocation to the nucleus. In this study, we examine the molecular mechanism by which RV P protein determines viral pathogenicity by comparing the IFN antagonist activities of the Nishigahara and Ni-CE P proteins. The results, obtained from both RV-infected cells and cells transfected to express P protein only, show that Ni-CE P protein is significantly impaired for its capacity to block IFN-activated STAT1 nuclear translocation and, consequently, inhibits IFN signaling less efficiently than Nishigahara P protein. Further, it was demonstrated that a defect in the nuclear export of Ni-CE P protein correlates with a defect in its ability to cause the mislocalization of STAT1. These data provide the first evidence that the capacity of the RV P protein to inhibit STAT1 nuclear translocation and IFN signaling correlates with the viral pathogenicity.
Mechanism of Initiation Site Selection Promoted by the Human Rhinovirus 2 Internal Ribosome Entry Site
Authors: Kaminski Ann , Pöyry Tuija A. A. , Skene Peter J. , Jackson Richard J.
Published: April 2010

Translation initiation site usage on the human rhinovirus 2 internal ribosome entry site (IRES) has been examined in a mixed reticulocyte lysate/HeLa cell extract system. There are two relevant AUG triplets, both in a base-paired hairpin structure (domain VI), with one on the 5′ side at nucleotide (nt) 576, base paired with the other at nt 611, which is the initiation site for polyprotein synthesis. A single residue was inserted in the apical loop to put AUG-576 in frame with AUG-611, and in addition another in-frame AUG was introduced at nt 593. When most of the IRES was deleted to generate a monocistronic mRNA, the use of these AUGs conformed to the scanning ribosome model: improving the AUG-576 context increased initiation at this site and decreased initiation at downstream sites, whereas the converse was seen when AUG-576 was mutated to GUA; and AUG-593, when present, took complete precedence over AUG-611. Under IRES-dependent conditions, by contrast, much less initiation occurred at AUG-576 than in a monocistronic mRNA with the same AUG-576 context, mutation of AUG-576 decreased initiation at downstream sites by ∼70%, and introduction of AUG-593 did not completely abrogate initiation at AUG-611, unless the apical base pairing in domain VI was destroyed by point mutations. These results indicate that ribosomes first bind at the AUG-576 site, but instead of initiating there, most of them are transferred to AUG-611, the majority by strictly linear scanning and a substantial minority by direct transfer, which is possibly facilitated by the occasional persistence of base pairing in the apical part of the domain VI stem.
The Virion Host Shutoff Endonuclease (UL41) of Herpes Simplex Virus Interacts with the Cellular Cap-Binding Complex eIF4F
Authors: Page Heidi G. , Read G. Sullivan
Published: April 2010

The herpes simplex virus Vhs endonuclease degrades host and viral mRNAs. Isolated Vhs cuts any RNA at many sites. Yet, within cells, it targets mRNAs and cuts at preferred sites, including regions of translation initiation. Previous studies have shown that Vhs binds the translation factors eIF4A and eIF4H. Here, we show that Vhs binds the cap-binding complex eIF4F. Association with eIF4F correlated with the ability of Vhs to bind eIF4A but not eIF4H. All Vhs proteins that degrade mRNAs associated with eIF4F. However, simply tethering an active endonuclease to eIF4F is not sufficient to degrade mRNAs. Binding to eIF4H may also be required.
Suppression of a Morphogenic Mutant in Rous Sarcoma Virus Capsid Protein by a Second-Site Mutation: a Cryoelectron Tomography Study
Authors: Butan Carmen , Lokhandwala Parvez M. , Purdy John G. , Cardone Giovanni , Craven Rebecca C. , Steven Alasdair C.
Published: April 2010

Retrovirus assembly is driven by polymerization of the Gag polyprotein as nascent virions bud from host cells. Gag is then processed proteolytically, releasing the capsid protein (CA) to assemble
de novo
inside maturing virions. CA has N-terminal and C-terminal domains (NTDs and CTDs, respectively) whose folds are conserved, although their sequences are divergent except in the 20-residue major homology region (MHR) in the CTD. The MHR is thought to play an important role in assembly, and some mutations affecting it, including the F167Y substitution, are lethal. A temperature-sensitive second-site suppressor mutation in the NTD, A38V, restores infectivity. We have used cryoelectron tomography to investigate the morphotypes of this double mutant. Virions produced at the nonpermissive temperature do not assemble capsids, although Gag is processed normally; moreover, they are more variable in size than the wild type and have fewer glycoprotein spikes. At the permissive temperature, virions are similar in size and spike content as in the wild type and capsid assembly is restored, albeit with altered polymorphisms. The mutation F167Y-A38V (referred to as FY/AV in this paper) produces fewer tubular capsids than wild type and more irregular polyhedra, which tend to be larger than in the wild type, containing ∼30% more CA subunits. It follows that FY/AV CA assembles more efficiently
in situ
than in the wild type and has a lower critical concentration, reflecting altered nucleation properties. However, its infectivity is lower than that of the wild type, due to a 4-fold-lower budding efficiency. We conclude that the wild-type CA protein sequence represents an evolutionary compromise between competing requirements for optimization of Gag assembly (of the immature virion) and CA assembly (in the maturing virion).
Glycosylation at 158N of the Hemagglutinin Protein and Receptor Binding Specificity Synergistically Affect the Antigenicity and Immunogenicity of a Live Attenuated H5N1 A/Vietnam/1203/2004 Vaccine Virus in Ferrets
Authors: Wang Weijia , Lu Bin , Zhou Helen , Suguitan Amorsolo L. , Cheng Xing , Subbarao Kanta , Kemble George , Jin Hong
Published: April 2010

A live attenuated influenza A/Vietnam/1203/2004 (H5N1) vaccine virus (VN04
ca
) has receptor binding specificity to α2,3-linked sialosides (α2,3SAL), and a single dose induces a minimal serum antibody response in mice and ferrets. In contrast, A/Hong Kong/213/2003 (H5N1) vaccine virus (HK03
ca
) binds to both α2,6SAL and α2,3SAL and generates a stronger serum antibody response in animals. Among the 9 amino acids that differed between the two H5 HA1 proteins, several HK03-specific residues enabled the VN04
ca
virus to bind to both α2,3SAL and α2,6SAL receptors, but only the removal of the 158N glycosylation, together with an S227N change, resulted in more-efficient viral replication in the upper respiratory tract of ferrets and an increased serum antibody response. However, the antibody response was HK03 strain specific and did not significantly cross-neutralize VN04 virus. A second approach was taken to adapt the H5N1 VN04
ca
virus in MDCK cells to select HA variants with larger plaque morphology. Although a number of large-plaque-size HA variants with amino acid changes in the HA receptor binding region were identified, none of these mutations affected virus receptor binding preference and immunogenicity. In addition, the known receptor binding site changes, Q226L and G228S, were introduced into the HA protein of the VN04
ca
virus. Only in conjunction with the removal of the 158N glycosylation did the virus replicate efficiently in the upper respiratory tract of ferrets and became more immunogenic, yet the response was also HK03 specific. Thus, the mask of the antigenic epitopes by 158N glycosylation at the HA globular head and its α2,3SAL binding preference of VN04
ca
virus affect virus antigenicity and replication in the host, resulting in a lower antibody response.
Regulation of Hepatitis C Virus Translation and Infectious Virus Production by the MicroRNA miR-122
Authors: Jangra Rohit K. , Yi MinKyung , Lemon Stanley M.
Published: April 2010

miR-122 is a liver-specific microRNA that positively regulates hepatitis C virus (HCV) RNA abundance and is essential for production of infectious HCV. Using a genetic approach, we show that its ability to enhance yields of infectious virus is dependent upon two miR-122-binding sites near the 5′ end of the HCV genome, S1 and S2. Viral RNA with base substitutions in both S1 and S2 failed to produce infectious virus in transfected cells, while virus production was rescued to near-wild-type levels in cells supplemented with a complementary miR-122 mutant. A comparison of mutants with substitutions in only one site revealed S1 to be dominant, as an S2 but not S1 mutant produced high virus yields in cells supplemented with wild-type miR-122. Translation of HCV RNA was reduced over 50% by mutations in either S1 or S2 and was partially rescued by transfection of the complementary miR-122 mutant. Unlike the case for virus replication, however, both sites function equally in regulating translation. We conclude that miR-122 promotes replication by binding directly to both sites in the genomic RNA and, at least in part, by stimulating internal ribosome entry site (IRES)-mediated translation. However, a comparison of the replication capacities of the double-binding-site mutant and an IRES mutant with a quantitatively equivalent defect in translation suggests that the decrement in translation associated with loss of miR-122 binding is insufficient to explain the profound defect in virus production by the double mutant. miR-122 is thus likely to act at an additional step in the virus life cycle.
Role for Complement in the Development of Seizures following Acute Viral Infection
Authors: Libbey Jane E. , Kirkman Nikki J. , Wilcox Karen S. , White H. Steve , Fujinami Robert S.
Published: April 2010

Complement, part of the innate immune system, acts to remove pathogens and unwanted host material. Complement is known to function in all tissues, including the central nervous system (CNS). In this study, we demonstrated the importance of the complement system within the CNS in the development of behavioral seizures following Theiler's murine encephalomyelitis virus (TMEV) infection. C57BL/6 mice, deficient in complement component C3, developed significantly fewer behavioral seizures following TMEV infection, whereas mice depleted of complement component C3 in the periphery through treatment with cobra venom factor had a seizure rate comparable to that of control mice. These studies indicate that C3 participates in the induction of acute seizures during viral encephalitis.
Full-Length Hepatitis B Virus Core Protein Packages Viral and Heterologous RNA with Similarly High Levels of Cooperativity
Authors: Porterfield J. Zachary , Dhason Mary Savari , Loeb Daniel D. , Nassal Michael , Stray Stephen J. , Zlotnick Adam
Published: April 2010

A critical feature of a viral life cycle is the ability to selectively package the viral genome.
In vivo
, phosphorylated hepatitis B virus (HBV) core protein specifically encapsidates a complex of pregenomic RNA (pgRNA) and viral polymerase; it has been suggested that packaging is specific for the complex. Here, we test the hypothesis that core protein has intrinsic specificity for pgRNA, independent of the polymerase. For these studies, we also evaluated the effect of core protein phosphorylation on assembly and RNA binding, using phosphorylated core protein and a phosphorylation mimic in which S155, S162, and S170 were mutated to glutamic acid. We have developed an
in vitro
system where capsids are disassembled and assembly-active core protein dimer is purified. With this protein, we have reassembled empty capsids and RNA-filled capsids. We found that core protein dimer bound and encapsidated both the HBV pregenomic RNA and heterologous RNA with high levels of cooperativity, irrespective of phosphorylation. In direct competition assays, no specificity for pregenomic RNA was observed. This suggests that another factor, such as the viral polymerase, is required for specific packaging. These results also beg the question of what prevents HBV core protein from assembling on nonviral RNA, preserving the protein for virus production.
β1Integrin Expression Increases Susceptibility of Memory B Cells to Epstein-Barr Virus Infection
Authors: Dorner Marcus , Zucol Franziska , Alessi Davide , Haerle Stephan K. , Bossart Walter , Weber Markus , Byland Rahel , Bernasconi Michele , Berger Christoph , Tugizov Sharof , Speck Roberto F. , Nadal David
Published: April 2010

Epstein-Barr virus (EBV) uses nasal mucosa-associated lymphoid tissue (NALT) as a portal of entry to establish life-long persistence in memory B cells. We previously showed that naïve and memory B cells from NALT are equally susceptible to EBV infection. Here we show that memory B cells from NALT are significantly more susceptible to EBV infection than those from remote lymphatic organs. We identify β1integrin, which is expressed the most by naïve B cells of distinct lymphoid origin and by memory B cells from NALT, as a mediator of increased susceptibility to infection by EBV. Furthermore, we show that BMRF-2-β1integrin interaction and the downstream signal transduction pathway are critical for postbinding events. An increase of β1integrin expression in peripheral blood memory B cells provoked by CD40 stimulation plus B-cell receptor cross-linking increased the susceptibility of non-NALT memory B cells to EBV infection. Thus, EBV seems to utilize the increased activation status of memory B cells residing in the NALT to establish and ensure persistence.
GENERATING FUNCTIONAL FOR GAUGE INVARIANT ACTIONS: EXAMPLES OF NONRELATIVISTIC GAUGE THEORIES
Authors: ANDREEV OLEG
Published: April 2010

We propose a generating functional for nonrelativistic gauge invariant actions. In particular, we consider actions without the usual magnetic term. Like in the Born–Infeld theory, there is an upper bound to the electric field strength in these gauge theories.
M1 TRANSITIONS OF MESONS IN A POTENTIAL MODEL OF INDEPENDENT QUARKS
Authors: JENA S. N. , MUNI M. K. , PATTNAIK H. R. , SAHU K. P.
Published: April 2010

The decay widths for M1 transitions of mesons are calculated in a relativistic potential model of independent quarks with its parameters determined from a fit to the mass splittings of the ground state mesons in the strange, charm and bottom flavor sector. A more realistic calculation of the decay widths is performed beyond static approximation incorporating some momentum dependence due to recoil of the daughter meson. The model accounts well for the observed decay widths of light mesons and predicts decay rates of heavy mesons which are comparable with those of other model calculations.
INFRARED BEHAVIOR OF THE ATTRACTIVE DI-QUARK CHANNEL IN THE QCD MODEL
Authors: KAWASHIMA KAZUHIRO
Published: April 2010

We examine the behavior of the attractive di-quark (qq) channel in the infrared region. Conventionally it is considered that the qq channel does not cause the fermion condensation in spite of its attractive interaction, because the quark field is dynamically massive due to the quark–antiquark condensation and the decoupling dynamics is applied. In this paper, we are going to reconsider rigorously the infrared behavior of the qq channel with paying attention to the stability of the effective action. And it is revealed that the behavior is not naively decided by the decoupling dynamics.
INVESTIGATING NON-FRITZSCH-LIKE TEXTURE SPECIFIC QUARK MASS MATRICES
Authors: MAHAJAN NEELU , VERMA ROHIT , GUPTA MANMOHAN
Published: April 2010

A detailed investigation of all possible textures of Fritzsch-like and non-Fritzsch-like, 144 for texture 6 zero and 432 for texture 5 zero mass matrices, have been carried out to ascertain their compatibility with the existing quark mixing data. It seems that all the texture 6 zero possibilities are completely ruled out whereas in the case of texture 5 zero mass matrices the only viable possibility looks to be that of Fritzsch-like.
ANALYSIS OF REGULAR INFLATIONARY COSMOLOGICAL MODELS WITH TWO TORSION FUNCTIONS IN POINCARÉ GAUGE THEORY OF GRAVITY
Authors: GARKUN A. S. , KUDIN V. I. , MINKEVICH A. V.
Published: April 2010

Regular inflationary Big Bang scenario with accelerating stage of cosmological expansion at asymptotics proposed earlier in the framework of the Poincaré gauge theory of gravity is investigated. Analysis of regular inflationary cosmological models with two torsion functions filled with scalar field with quadratic potential and ultrarelativistic matter is carried out numerically. Properties of different stages of regular inflationary cosmological solutions are studied, restrictions on admissible values of parameters and initial conditions at a bounce are found. The structure of extremum surface in space of physical variables is investigated.
Two residues in the extracellular domain convert a nonfunctional ASIC1 into a proton-activated channel
Authors: Li Tianbo , Yang Youshan , Canessa Cecilia M.
Published: April 2010

Acid-sensing ion channels (ASICs) are proton-activated sodium channels of the nervous system. Mammals express four ASICs, and orthologs of these genes have been found in all chordates examined to date. Despite a high degree of sequence conservation of all ASICs across species, the response to a given increase in external proton concentration varies markedly: from large and slowly inactivating inward currents to no detectable currents. The underlying bases of this functional variability and whether it stems from differences in proton-binding sites or in structures that translate conformational changes have not been determined yet. We show here that the ASIC1 ortholog of an early vertebrate, lamprey ASIC1, does not respond to protons; however, only two amino acid substitutions for the corresponding ones in rat ASIC1, Q77L and T85L, convert lamprey ASIC1 into a highly sensitive proton-activated channel with apparent H+ affinity of pH50 7.2. Addition of C73H increases the magnitude of the currents by fivefold, and W64R confers desensitization similar to that of the mammalian counterpart. Most amino acid substitutions in these four positions increase the rates of opening and closing the pore, whereas only few, namely, the ones in rat ASIC1, slow the rates. The four residues are located in a contiguous segment made by the β1-β2-linker, β1-strand, and the external segment of the first transmembrane helix. We conclude that the segment thus defined modulates the kinetics of opening and closing the pore and that fast kinetics of desensitization rather than lack of acid sensor accounts for the absence of proton-induced currents in the parent lamprey ASIC1.
Maternal obesity and overnutrition alter fetal growth rate and cotyledonary vascularity and angiogenic factor expression in the ewe
Authors: Ma Yan , Zhu Mei J. , Zhang Liren , Hein Sarah M. , Nathanielsz Peter W. , Ford Stephen P.
Published: April 2010

In pregnant sheep, maternal:fetal exchange occurs across placentomes composed of placental cotyledonary and uterine caruncular tissues. Recently, we reported that fetal weights of obese (OB) ewes [fed a diet of 150% of National Research Council (NRC) recommendations] were ∼30% greater than those of control (C) ewes (fed a diet 100% of NRC recommendations) at midgestation (MG), but fetal weights were similar in late gestation (LG). Transplacental nutrient exchange is dependent on placental blood flow, which itself is dependent on placental vascularity. The current study investigated whether the observed initial faster and subsequent slower fetal growth rate of OB compared with C was associated with changes in cotyledonary vascularity and expression of angiogenic factors (vascular endothelial growth factor, fibroblast growth factor-2, placental growth factor, angiopoietin-1 and -2). Cotyledonary arteriole diameters were markedly greater ( P < 0.05) in OB than C ewes at MG, but while arteriole diameter of C ewes increased ( P < 0.05) from MG to LG, they remained unchanged in OB ewes. Cotyledonary arterial angiogenic factors mRNA and protein expression were lower ( P < 0.05) in OB than C ewes at MG and remained low from MG to LG. In contrast, mRNA levels of angiogenic factors in C ewes declined from high levels at MG to reach those of OB ewes by LG. The increase in cotyledonary arteriole diameter in early to MG may function to accelerate fetal growth rate in OB ewes, while the decreased cotyledonary arterial angiogenic factors from MG-LG may function to protect the fetus from excessive placental vascular development, increased maternal nutrient delivery, and excessive weight gain.
Tidal stretches do not modulate responsiveness of intact airways in vitro
Authors: LaPrad Adam S. , Szabo Thomas L. , Suki Béla , Lutchen Kenneth R.
Published: April 2010

Studies on isolated tracheal airway smooth muscle (ASM) strips have shown that length/force fluctuations, similar to those likely occurring during breathing, will mitigate ASM contractility. These studies conjecture that, solely by reducing length oscillations on a healthy, intact airway, one can create airway hyperresponsiveness, but this has never been explicitly tested. The intact airway has additional complexities of geometry and structure that may impact its relevance to isolated ASM strips. We examined the role of transmural pressure (Ptm) fluctuations of physiological amplitudes on the responsiveness of an intact airway. We developed an integrated system utilizing ultrasound imaging to provide real-time measurements of luminal radius and wall thickness over the full length of an intact airway ( generation 10 and below) during Ptm oscillations. First, airway constriction dynamics to cumulative acetylcholine (ACh) doses (10−7 to 10−3 M) were measured during static and dynamic Ptm protocols. Regardless of the breathing pattern, the Ptm oscillation protocols were ineffective in reducing the net level of constriction for any ACh dose, compared with the static control ( P = 0.225–0.793). Next, Ptm oscillations of increasing peak-to-peak amplitude were applied subsequent to constricting intact airways under static conditions (5.0-cmH2O Ptm) with a moderate ACh dose (10−5 M). Peak-to-peak Ptm oscillations ≤5.0 cmH2O resulted in no statistically significant bronchodilatory response ( P = 0.429 and 0.490). Larger oscillations (10 cmH2O, peak to peak) produced modest dilation of 4.3% ( P = 0.009). The lack of modulation of airway responsiveness by Ptm oscillations in intact, healthy airways suggests that ASM level mechanisms alone may not be the sole determinant of airway responsiveness.
Neonatal Sensory Deprivation and the Development of Cortical Function: Unilateral and Bilateral Sensory Deprivation Result in Different Functional Outcomes
Authors: Popescu Maria V. , Ebner Ford F.
Published: April 2010

The normal development of sensory perception in mammals depends on appropriate sensory experience between birth and maturity. Numerous reports have shown that trimming some or all of the large mystacial vibrissa (whiskers) on one side of the face after birth has a detrimental effect on the maturation of cortical function. The objective of the present study was to understand the differences that occur after unilateral whisker trimming compared with those that occur after bilateral deprivation. Physiological deficits produced by bilateral trimming (BD) of all whiskers for 2 mo after birth were compared with the deficits produced by unilateral trimming (UD) for the same period of time using extracellular recording under urethan anesthesia from single cells in rat barrel cortex. Fast spiking (FSUs) and regular spiking (RSUs) units were separated and their properties compared in four subregions identified by histological reconstructions of the electrode penetrations, namely: layer IV barrel and septum, and layers II/III above a barrel and above a septum. UD upregulated responses in layer IV septa and in layers II/III above septa and perturbed the timing of responses to whisker stimuli. After BD, nearly all responses were decreased, and poststimulus latencies were increased. Circuit changes are proposed as an argument for how inputs arising from the spared whiskers project to the undeprived cortex and, via commissural fibers, could upregulate septal responses after UD. Following BD, more global neural deficits create a signature difference in the outcome of UD and BD in rat barrel cortex.
Investigating Neural Correlates of Behavior in Freely Behaving Rodents Using Inertial Sensors
Authors: Venkatraman Subramaniam , Jin Xin , Costa Rui M. , Carmena Jose M.
Published: April 2010

Simultaneous behavior and multielectrode neural recordings in freely behaving rodents holds great promise to study the neural bases of behavior and disease models in combination with genetic manipulations. Here, we introduce the use of three-axis accelerometers to characterize the behavior of rats and mice during chronic neural recordings. These sensors were small and light enough to be worn by rodents and were used to record three-axis acceleration during freely moving behavior. A two-layer neural network-based pattern recognition algorithm was developed to extract the natural behavior of mice from the acceleration data. Successful recognition of resting, eating, grooming, and rearing are shown using this approach. The inertial sensors were combined with continuous 24-h recordings of neural data from the striatum of mice to characterize variations in neural activity with circadian cycles and to study the neural correlates of spontaneous action initiation. Finally, accelerometers were used to study the performance of rodents in traditional operant conditioning, where they were used to extract the reaction time of rodents. Thus the addition of accelerometer recordings of rodents to chronic multielectrode neural recordings provides great value for a number of neuroscience applications.
Self-Reported Psychosocial Health in Obese Patients before and after Weight Loss
Authors: Osei-Assibey G. , Kyrou I. , Kumar S. , Saravanan P. , Matyka K. A.
Published: April 2010

Psychosocial profiles were examined in 255 morbidly obese patients attending a hospital service offering access to standard weight loss therapies. 129 patients were reassessed after at least 6-month follow-up. At baseline, 51.8% and 32.7% of patients, respectively, had evidence of anxiety and depressive disorders, 24% had severe impairments in self esteem, and 29.7% had an increased risk of eating disorders. At follow-up, weight loss from baseline was significant in all 3 therapies: diet only is  kg; pharmacotherapy is  kg; and surgery is  kg. Anxiety scores improved in all three groups (). Patients having pharmacotherapy or surgery had significant improvements in physical and work function and public distress compared to those having dietary treatment only (). Our observational data suggest that weight management services can lead to psychosocial benefit in morbidly obese patients. Well-designed studies are necessary to examine the link between weight loss and emotional health.
WiFi and WiMAX Secure Deployments
Authors: Trimintzios Panagiotis , Georgiou George
Published: April 2010

Wireless Broadband offers incredibly fast, “always on” Internet similar to ADSL and sets the user free from the fixed access areas. In order to achieve these features standardisation was achieved for Wireless LAN (WLANs) and Wireless Metropolitan Area Networks (WMANs) with the advent of IEEE802.11 and IEEE802.16 family of standards, respectively. One serious concern in the rapidly developing wireless networking market has been the security of the deployments since the information is delivered freely in the air and therefore privacy and integrity of the transmitted information, along with the user-authentication procedures, become a very important issue. In this article, we present the security characteristics for the WiFi and the WiMAX networks. We thoroughly present the security mechanisms along with a threat analysis for both IEEE 802.11 and the 802.16 as well as their amendments. We summarise in a comparative manner the security characteristics and the possible residual threats for both standards. Finally focus on the necessary actions and configurations that are needed in order to deploy WiFi and WiMAX with increased levels of security and privacy.
Nest Relocation and Colony Founding in the Australian Desert Ant,Melophorus bagotiLubbock (Hymenoptera: Formicidae)
Authors: Schultheiss Patrick , Schwarz Sebastian , Wystrach Antoine
Published: April 2010

Even after years of research on navigation in the Red Honey Ant,Melophorus bagoti, much of its life history remains elusive. Here, we present observations on nest relocation and the reproductive and founding stages of colonies. Nest relocation is possibly aided by trail laying behaviour, which is highly unusual for solitary foraging desert ants. Reproduction occurs in synchronised mating flights, which are probably triggered by rain. Queens may engage in multiple matings, and there is circumstantial evidence that males are chemically attracted to queens. After the mating flight, the queens found new colonies independently and singly. Excavation of these founding colonies reveals first insights into their structure.
A Pivotal Role for Interleukin-27 in CD8+T Cell Functions and Generation of Cytotoxic T Lymphocytes
Authors: Morishima Noriko , Mizoguchi Izuru , Okumura Masae , Chiba Yukino , Xu Mingli , Shimizu Motomu , Matsui Masanori , Mizuguchi Junichiro , Yoshimoto Takayuki
Published: April 2010

Cytotoxic T lymphocytes (CTLs) play a critical role in the control of various cancers and infections, and therefore the molecular mechanisms of CTL generation are a critical issue in designing antitumor immunotherapy and vaccines which augment the development of functional and long-lasting memory CTLs. Interleukin (IL)-27, a member of the IL-6/IL-12 heterodimeric cytokine family, acts on naiveCD4+T cells and plays pivotal roles as a proinflammatory cytokine to promote the early initiation of type-1 helper differentiation and also as an antiinflammatory cytokine to limit the T cell hyperactivity and production of pro-inflammatory cytokines. Recent studies revealed that IL-27 plays an important role inCD8+T cells as well. Therefore, this article reviews current understanding of the role of IL-27 inCD8+T cell functions and generation of CTLs.
FSHRSingle Nucleotide Polymorphism Frequencies in Proven Fathers and Infertile Men in Southeast Turkey
Authors: Balkan Mahmut , Gedik Abdullah , Akkoc Hasan , Izci Ay Ozlem , Erdal M. Emin , Isi Hilmi , Budak Turgay
Published: April 2010

The influence of FSH receptor (FSHR) variants on male infertility is not completely understood. The present investigation is the first screening study for SNP at nucleotide position −29 in the core promoter region and codon 680 in exon 10 of theFSHRand the effect of the serum levels of FSH on male infertility in Southeast Turkey. The SNPs in codon 680 and at position −29 of theFSHRgene were analyzed by PCR-RFLP technique in 240 men with proven fathers, and 270 infertile men (150 nonobstructive azoospermic and 120 severe oligozoospermic). The separate analysis for SNP at nucleotide position −29 did not show any difference in genotypic frequencies and serum FSH levels. The genotype distribution of SNP at position 680 was different but does not influence serum FSH levels. Together the two SNPs form four discrete haplotypes (A-Thr-Asn, G-Thr-Asn, A-Ala-Ser, and G-Ala-Ser) occurring in 10 combinations. A statistically significant difference in the allelic distribution of G-Asn/G-Ser and G-Ser/G-Ser genotype between proven fathers and infertile men but there were not any statistically significant difference in the overall frequency of the four FSHR haplotypes. We conclude that the FSHR haplotype does not associate with different serum FSH levels but it is differently distributed in proven fathers and infertile men.
CATHARE Assessment of PACTEL LOCA Experiments with Accident Management
Authors: Sabotinov Luben , Purhonen Heikki , Riikonen Vesa
Published: April 2010

This paper summarizes the analysis results of three PACTEL experiments, carried out with the advanced thermal-hydraulic system computer CATHARE 2 code as a part of the second work package WP2 (analytical work) of the EC project “Improved Accident Management of VVER nuclear power plants” (IMPAM-VVER). The three LOCA experiments, conducted on the Finnish test facility PACTEL (VVER-440 model), represent 7.4% cold leg breaks with combination of secondary bleed and primary bleed and feed and different actuation modes of the passive safety injection. The code was used for both defining and analyzing the experiments, and to assess its capabilities in predicting the associated complex VVER-related phenomena. The code results are in reasonable agreement with the measurements, and the important physical phenomena are well predicted, although still further improvement and validation might be necessary.
Optical Properties of Al- and Sb-Doped CdTe Thin Films
Authors: Al-Douri A. A. J. , Al-Shakily F. Y. , Alias M. F. A. , Alnajjar A. A.
Published: April 2010

Nondoped and (Al, Sb)-doped CdTe thin films with 0.5, 1.5, and 2.5  wt.%, respectively, were deposited by thermal evaporation technique under vacuum onto Corning 7059 glass at substrate temperatures () of room temperature (RT) and 423 K. The optical properties of deposited CdTe films such as band gap, refractive index (n), extinction coefficient (), and dielectric coefficients were investigated as function of Al and Sb wt.% doping, respectively. The results showed that films have direct optical transition. Increasing and the wt.% of both types of dopant, the band gap decrease but the optical is constant asn, and real and imaginary parts of the dielectric coefficient increase.
Anticoagulant therapy in critical organ ischaemia/reperfusion injury
Authors: Cate Hugo ten , Spronk Henri M. H. , Loubele Sarah T. B. G.
Published: April 2010

Ischaemia/reperfusion (I/R) injury is central to a number of pathologies including myocardial infarction and stroke. Several cellular processes are involved in the progress of I/R injury, involving complex interactions between coagulation and inflammatory or apoptotic processes. Besides for their anti-coagulant function, anticoagulant proteins such as activated protein C (APC), active site inhibited factor VIIa (ASIS), tissue factor pathway inhibitor (TFPI), and antithrombin (AT) are also known for their anti-inflammatory or cell protective effects. This review gives an overview of the application of these anti-coagulants in several animal models of I/R injury in critical organs and describes the effects of these proteins on cellular processes including inflammation and apoptosis. The future testing of mutant forms of some of these inhibitors including APC in a clinical setting should be actively explored.
Factor VIIa-antithrombin complexes in patients with arterial and venous thrombosis
Authors: Spiezia Luca , Rossetto Valeria , Campello Elena , Gavasso Sabrina , Woodhams Barry , Tormene Daniela , Simioni Paolo
Published: April 2010

Antithrombin (AT), in the presence of heparin, is able to inhibit the catalytic activity of factor VIIa bound to tissue factor (TF) on cell surfaces. The clinical meaning of FVIIa-AT complexes plasma levels is unknown. It was the objective of this study to evaluate FVIIa-AT complexes in subjects with thrombosis. Factor VIIa-AT complexes plasma levels in 154 patients consecutively referred to our Department with arterial or venous thrombosis and in a group of 154 healthy subjects, were measured. Moreover, FVIIa-AT complexes were determined in: i) n = 53 subjects belonging to 10 families with inherited factor VII deficiency; ii) n = 58 subjects belonging to seven families with AT deficiency; iii) n = 49 patients undergoing oral anticoagulant therapy (OAT). Factor VIIaAT levels were determined by a specific ELISA kit (R&D, Diagnostica Stago, Gennevilliers, France). Factor VIIa-AT complexes mean plasma levels were lower in patients with either acute arterial (136 ± 40 pM) or venous (142 ± 53 pM) thrombosis than subjects with previous thrombosis (arterial 164 ± 33 pM and venous 172 ± 61 pM, respectively) and than healthy controls (156 ± 63 pM). Differences between acute and previous thrombosis, were statistically significant (p < 0.05). Subjects with inherited and acquired (under OAT) factor VII deficiency had statistically significant lower FVIIa-AT complexes plasma levels (80 ± 23 pM and 55 ± 22 pM, respectively) than controls (150 ± 51 pM, p < 0.0001 and 156 ± 63 pM, p < 0.00001, respectively). Factor VIIa-AT complexes are positively correlated with plasma factor VII/VIIa levels. Further investigations are needed to verify the possible role of higher FVIIa-AT complex plasma levels in predicting hypercoagulable states and thrombosis.
Relationship between degree of P2Y12 receptor blockade and inhibition of P2Y12-mediated platelet function
Authors: Buckland Robert , Sugidachi Atsuhiro , Jakubowski Joseph , Storey Robert , Judge Heather
Published: April 2010

The thienopyridine P2Y12 receptor antagonists clopidogrel and prasugrel prevent arterial thrombosis and are routinely used following percutaneous coronary intervention. However, the optimal level of P2Y12 blockade to effectively inhibit platelet function is unknown. These studies utilised the active metabolite of prasugrel (R-138727) to achieve a range of P2Y12 blockade in vitro and assessed several aspects of platelet function. Blood from healthy volunteers was incubated with R-138727 (0–10 μM). P2Y12 receptor number was assessed using a 33P-2MeSADP binding assay. Platelet aggregation (PA) was measured by optical aggregometry with ADP 2–20 μM. VASP phosphorylation, annexin V binding, microparticle formation and P-selectin expression were assessed by flow cytometry. Increasing numbers of unblocked receptors were required for a sustained aggregation response with decreasing concentrations of ADP. A P2Y12 receptor blockade of 60–80% resulted in strong inhibition of final PA response, P-selectin expression, microparticle formation and vasodilator-stimulated phosphoprotein (VASP). PA induced by ADP 2 μM and P-selectin expression were particularly sensitive to low levels of receptor blockade whereas the VASP phosphorylation assay was relatively insensitive, requiring 60% receptor blockade to achieve substantial inhibition. Different assays varied in their ability to discriminate particular ranges of P2Y12 blockade and 80% or greater P2Y12 receptor blockade is required for consistently strong inhibition of several aspects of platelet function. These data guide the interpretation of results from different assays used to monitor the effects of P2Y12 receptor antagonists.
The role of thrombin and protease-activated receptors in pain mechanisms
Authors: García Paul , Gulati Amitabh , Levy Jerrold
Published: April 2010

As our knowledge of the mechanisms underlying the sensation of pain continues to expand, researchers are constantly searching for novel therapeutic targets. One such novel pain pathway involves thrombin and its associated protease-activated receptor (PAR). Besides its traditional role in haemostasis, thrombin has multiple roles in both the central and peripheral nervous system including activation of microglia, regulation of neuronal death and neurite outgrowth, and influencing the transmission of pain signals in the nociceptive circuitry. Eventually therapeutic modalities directed at these targets could provide novel therapeutic approaches for treating chronic pain. The thrombin-associ-ated PARs also have roles in inflammation, neurodevelopment, and conducting pain, both in conjunction with thrombin and independently. Recent laboratory evidence suggests that the PARs can attenuate pain mediated by the enteric nervous system in animal models (for example in pancreatitis and colitis). This review highlights several pathways in the mediation of pain sensation that can be influenced by thrombin.
Validation of the Health ABC Heart Failure Model for Incident Heart Failure Risk Prediction
Authors: Kalogeropoulos Andreas , Psaty Bruce M. , Vasan Ramachandran S. , Georgiopoulou Vasiliki , Smith Andrew L. , Smith Nicholas L. , Kritchevsky Stephen B. , Wilson Peter W.F. , Newman Anne B. , Harris Tamara B. , Butler Javed ,   for the Cardiovascular Health Study
Published: April 2010

Background—

The recently developed and internally validated Health ABC HF model uses 9 routinely available clinical variables to determine incident heart failure risk. In this study, we sought to externally validate the Health ABC HF model.

Methods and Results—

Observed 5-year incidence of heart failure, defined as first hospitalization for new-onset heart failure, was compared with 5-year risk estimates derived from the Health ABC HF model among participants without heart failure at baseline in the Cardiovascular Health Study. During follow-up, 400 of 5335 (7.5%) participants developed heart failure over 5 years versus 364 (6.8%) predicted by the Health ABC HF model (predicted-to-observed ratio, 0.90). Observed versus predicted 5-year heart failure probabilities were 3.2% versus 2.8%, 9.0% versus 7.0%, 15.9% versus 13.7%, and 24.6% versus 30.8% for the <5%, 5% to 10%, 10% to 20%, and >20% 5-year risk categories, respectively. The Hosmer-Lemeshow χ
2
was 14.72 (degrees of freedom, 10;
P
=0.14), and the C index was 0.74 (95% CI, 0.72 to 0.76). Calibration and discrimination demonstrated adequate performance across sex and race overall; however, risk was underestimated in white men, especially in the 5% to 10% risk category. Model performance was optimal when participants with normal left ventricular function at baseline were assessed separately. Performance was consistent across age groups. Analyses with death as a competing risk yielded similar results.

Conclusions—

The Health ABC HF model adequately predicted 5-year heart failure risk in a large community-based study, providing support for the external validity of the model. This tool may be used to identify individuals to whom to target heart failure prevention efforts.
Public/Private Partnerships in the Public Health Sector
Authors: Burci Gian Luca
Published: April 2010

This article reviews and analyses the growing phenomenon of public-private partnerships in the field of international public health cooperation (PPPs). This trend has been led by the changing political dynamics surrounding international health issues as well as by the perception of the inadequacy of market mechanisms and the inability of existing international institutions for tackling pressing health problems in developing countries. The article analyses in particular the remarkable developments in international health governance that PPPs have ushered in as well as the role played and the challenges faced by the World Health Organization.
Mandate Issues in the Activities of the International Fund for Agricultural Development (IFAD)
Authors: Martha Rutsel Silvestre
Published: April 2010

Questions of mandate are central in the actual operations of intergovernmental organizations within secretariats, in opinions of legal counsel, and in governing councils and general assemblies. Mandate issues can impose real constraints, or generate demands for action, or be brushed aside in some political circumstances. Overall they are a significant and perplexing part of the administrative law of international organizations. This paper explores the highly varied practical effect of mandate issues on operations of an international organization, through analysis of diverse approaches to mandate constraints and aspirations in ventures of the International Fund for Agricultural Development (IFAD).
Promoting Human Rights in the Administration of Justice in Southern Sudan. Mandate and Accountability Dilemmas in the Fiel Work of a DPKO Human Rights Officer
Authors: Garms Ulrich
Published: April 2010

The UNMIS Human Rights Section is called to promote respect for international human rights standards as part of a peacekeeping operation in a post-conflict society. As such, it is exposed to conflicting but equally legitimate demands from different stakeholders. To illustrate some of the dilemmas arising in practice from the tensions between these demands, the paper looks at three case studies taken from the work of the UNMIS Human Rights Section in Southern Sudan. They concern the tension between customary law and the protection of women's rights, the right to counsel in capital cases, and justice for atrocities committed during the civil war. The paper argues that, also because of the inherent fundamental contradictions in what a field presence such as the UNMIS Human Rights Section seeks to achieve, attempts to promote meaningful accountability of the field operation for the results obtained encounter significant limitations.