bluuebunny/crossref_metadata_2025 huggingface dataset:
Stability of Copigmented Anthocyanins and Ascorbic Acid in Muscadine Grape Juice Processed by High Hydrostatic Pressure
Authors: Del Pozo‐Insfran D. , Del Follo‐Martinez A. , Talcott S.T. , Brenes C. H.
Published: April 2007

ABSTRACT:  Intermolecular copigmentation is one of the mechanisms of stabilization of anthocyanins in nature and is also responsible for the characteristic color and stability of aged red wines. In the present study, the effect of polyphenol oxidase (PPO) activity on phytochemical stability of an ascorbic acid‐fortified muscadine grape juice following high hydrostatic pressure (HHP) processing (400 and 550 MPa for 15 min) and after 21 d of storage at 25 °C was investigated. Addition of rosemary and thyme polyphenolic extracts (copigmentation) was evaluated as a means to stabilize anthocyanins and ascorbic acid during pressurization and subsequent storage. Polyphenolic extracts were partially purified in order to reduce their content of PPO substrates, and improve their stabilization properties within juice matrix. Overall PPO activity increased (3‐ and 2.5‐fold) following HHP at 400 and 550 MPa, respectively, although it was significantly lower in copigmented treatments. Higher anthocyanin losses occurred at 400 (∼70%) than at 550 MPa (∼46%), which were correlated to antioxidant losses (r= 0.89). Similarly, greater ascorbic acid losses were observed at 400 (84%) than at 550 MPa (18%). Copigmentation increased anthocyanin retention in reference to pressurized controls (3‐ and 3.2‐fold for rosemary and thyme treatments, respectively) and decreased ascorbic degradation (20 to 32%). In stored samples, higher anthocyanin content (>2‐fold) and antioxidant capacity (>1.5‐fold) was observed for copigmented treatments when compared to control juices. Addition of partially purified copigments increased muscadine grape juice color, antioxidant activity and also reduced phytochemical losses during HHP processing and storage.
2‐Step Optimization of the Extraction and Subsequent Physical Properties of Channel Catfish (Ictalurus punctatus) Skin Gelatin
Authors: Yang Hongshun , Wang Yifen , Jiang Mingkang , Oh Jun‐Hyun , Herring Josh , Zhou Peng
Published: April 2007

ABSTRACT:  To optimize the extraction of gelatin from channel catfish (Ictalurus punctatus) skin, a 2‐step response surface methodology involving a central composite design was adopted for the extraction process. After screening experiments, concentration of NaOH, alkaline pretreatment time, concentration of acetic acid, and extraction temperature were selected as the independent variables. In the 1st step of the optimization the dependent variables were protein yield (YP), gel strength (GS), and viscosity (V). Seven sets of optimized conditions were selected from the 1st step for the 2nd‐step screen. Texture profile analysis and the 3 dependent variables from the 1st step were used as responses in the 2nd‐step optimization. After the 2nd‐step optimization, the most suitable conditions were 0.20 M NaOH pretreatment for 84 min, followed by a 0.115 M acetic acid extraction at 55 °C. The optimal values obtained from these conditions were YP = 19.2%, GS = 252 g, and V = 3.23 cP. The gelatin obtained also showed relatively good hardness, cohesiveness, springiness, and chewiness. The yield of protein and viscosity can be predicted by a quadratic and a linear model, respectively.
Reuse of Ozonated Alkaline Solutions as Fermentation Brines in Spanish Green Table Olives
Authors: Segovia‐Bravo K.A. , Arroyo‐López F.N. , García‐García P. , Durán‐Quintana M.C. , Garrido‐Fernández A.
Published: April 2007

ABSTRACT:  The water used to reduce the excess NaOH after the immersion of green olives in lye becomes a heavily polluted, alkaline wastewater (AW) once it has come into contact with the fruits. Its treatment with ozonated air for 72 h (TAW) destroyed all polyphenols in the solution. A comparison of the microbial, physicochemical, and organoleptic characteristics of olives processed in the traditional way (W) and those reusing AW or TAW was made. The reuse of TAW or diluted TAW (TAW + W) as fermentation brines led to a shorter lag phase, higher maximum specific growth of lactobacillus, and higher lactic acid accumulation (in TAW) than the traditional process; differences among other physicochemical characteristics were not relevant, except that reused brines (AW) always had higher polyphenol contents. The organoleptic panel test did not detect significant differences among treatments in acidic taste, firmness, pit detachment, or olive color. Overall quality was fairly similar for olives from the traditional process and those from the reused TAW. Direct (or diluted) reuse of AW was also possible but produced a more bitter tasting olive.
A Comparison of Nutrient Density Scores for 100% Fruit Juices
Authors: Rampersaud G.C.
Published: April 2007

ABSTRACT:  The 2005 Dietary Guidelines for Americans recommend that consumers choose a variety of nutrient‐dense foods. Nutrient density is usually defined as the quantity of nutrients per calorie. Food and nutrition professionals should be aware of the concept of nutrient density, how it might be quantified, and its potential application in food labeling and dietary guidance. This article presents the concept of a nutrient density score and compares nutrient density scores for various 100% fruit juices. One hundred percent fruit juices are popular beverages in the United States, and although they can provide concentrated sources of a variety of nutrients, they can differ considerably in their nutrient profiles. Six methodologies were used to quantify nutrient density and 7 100% fruit juices were included in the analysis: apple, grape, pink grapefruit, white grapefruit, orange, pineapple, and prune. Food composition data were obtained from the USDA National Nutrient Database for Standard Reference, Release 18. Application of the methods resulted in nutrient density scores with a range of values and magnitudes. The relative scores indicated that citrus juices, particularly pink grapefruit and orange juice, were more nutrient dense compared to the other nonfortified 100% juices included in the analysis. Although the methods differed, the relative ranking of the juices based on nutrient density score was similar for each method. Issues to be addressed regarding the development and application of a nutrient density score include those related to food fortification, nutrient bioavailability, and consumer education and behavior.
Effect of UV Irradiation on Cut Cantaloupe: Terpenoids and Esters
Authors: Beaulieu John C.
Published: April 2007

ABSTRACT: Recent studies demonstrated that ultraviolet (UV) radiation enhanced terpenoids and decreased esters in thin‐sliced cantaloupe. In preliminary studies treating fresh juices with UV, terpenoid compounds that normally were not isolated, or found in minute quantities, were elevated and only low molecular weight alcohols, ketones, and aldehydes decreased. Subsequently terpenoid induction/oxidation in UV‐treated cut cantaloupe was reinvestigated. UV exposure increased the concentrations of terpenoids in cantaloupe tissue. However, UV exposure alone was not the sole factor responsible for enhanced terpenoids. UV‐enhanced terpenoid production appears to be both cultivar‐ and maturity‐dependent. Concomitant decreases in the ester content of UV‐treated samples occurred using a previously published system. Yet, we established that almost identical ester losses occurred in thinly sliced laminar tissue receiving 60 min UV or air exposure in an open system. Tissue samples that were exposed to UV in a closed system often did not suffer correspondingly equal ester loss. Marked tissue warming (4.3 ± 0.5 °C in 60 min) occurred during UV treatments in thin‐sliced tissue. Ester loss from cantaloupe tissue was caused by the experimental procedure, but not by UV treatmentper se. These findings are supported by the observation that UV is not responsible for chemical transformations to ester bonds, esterase, and lipase decrease in stored cut cantaloupe, and no lipid oxidation volatiles were observed in thin‐sliced control tissue, while oxidized terpenoids were recovered. Information gathered indicates that improper cutting, handling, sanitation treatment, and storage can radically alter the desirable volatile aroma profile in cut cantaloupe, and potentially lead to decreased consumer acceptance.
Ethical stockmanship
Authors: Hemsworth PH
Published: April 2007

The objective of this review is to consider the ethics of stockmanship, particularly from the perspective of the nature and extent of the duties of stockpeople to their farm animals.

It will consider what science tells us about the impact of stockmanship on the animal, particularly the welfare of the farm animal. The effects of human–animal interactions on the stockperson will also be considered, since these interactions affect the work performance and job satisfaction of the stockperson and thus indirectly affect animal welfare. Animal ethics is broader than animal welfare and includes economic as well as philosophical, social, cultural and religious aspects. This paper is predicated on the view that farm animals can suffer, and that animal suffering is a key consideration in our moral obligations to animals.

Housing and husbandry practices affect farm animal welfare and thus farmers and stockpeople have a responsibility to provide, at minimum, community‐acceptable animal housing and husbandry standards for their animals. The farmer's or stockperson's attitudes and behaviour can directly affect the animal's welfare and thus they also have a responsibility to provide specific standards of stockmanship for these animals. However, research suggests that the behaviour of some stockpeople is not as correct as it might be. Such situations exemplify the inevitably unequal human – domestic animal relationship, and this inequality should be considered in analysing the boundary between right and wrong behaviour of humans. Thus ethical discussion, using science and other considerations and involving stockpeople, livestock industries, government and the general public, should be used to establish and assure acceptable stockperson competencies across the livestock industries. Training programs targeting the key attitudes and behaviour of stockpeople presently offer the livestock industries good opportunities to improve human–animal interactions.
Pimelea trichostachya poisoning (St George disease) in horses
Authors: Wilson S‐J , Taylor JD , Gibson JA , Mckenzie RA
Published: April 2007

A dense population of Pimelea trichostachya plants (Family Thymelaeaceae) in pasture poisoned a horse herd in southern inland Queensland in October–November 2005. Plant density was 2 to 45 g wet weight/m2 (mean 16 g/m2) from 5 to 69 plants/m2 (mean 38 plants/m2) representing 3 to 20% (mean 9%) of the volume of pasture on offer. Ten of 35 mares, fillies and geldings were affected. Clinical signs were loss of body weight, profound lethargy, serous nasal discharge, severe watery diarrhoea and subcutaneous oedema of the intermandibular space, chest and ventral midline. Pathological findings were anaemia, leucocytopenia, hypoproteinaemia, dilatation of the right ventricle of the heart, dilated hepatic portal veins and periportal hepatic sinusoids (peliosis hepatis), alimentary mucosal hyperaemia and oedema of mesenteric lymph nodes. Cattle grazing the same pasture were affected by Pimelea poisoning simultaneously. Removal of the horses to Pimelea‐free pasture initiated recovery. The one other incident of this syndrome, previously only recognised in cattle in Australia, occurred in horses, in South Australia in 2002, with access to a dense Pimelea simplex population.
Ganglioneuritis causing high mortalities in farmed Australian abalone (Haliotis laevigata and Haliotis rubra)
Authors: Hooper C , Hardy‐Smith P , Handlinger J
Published: April 2007

Objective  To investigate an outbreak of sudden severe mortality in farmed abalone from coastal Victoria.

Results  The outbreaks occurred almost simultaneously in three farms following abalone movements from the wild and between farms. The initial on farm investigation identified a number of features that when considered together were highly suggestive of an infectious aetiology. In many cases, dead abalone had no significant gross lesions. Others had swollen mouths and some had prolapse and eversion of the radula. Histologically, the lesions centred on the nerves innervating the labial apparatus, primarily the cerebral and buccal ganglia, cerebral commissure and peripheral nerve branches arising from these. Nervous tissue necrosis and haemocyte infiltration were the dominant lesions seen microscopically in affected nerves.

Conclusions  A recent outbreak of mortality in Australian abalone was associated with neurotropic lesions, which have not previously been described in this country. The on farm and between farm pattern of spread of the outbreak, a history of abalone movements linking farms, clinical observation of moribund and dead abalone were all highly suggestive of a virulent infectious agent.
Maintenance Priority Assignment Utilizing On-line Production Information
Authors: Yang Zimin , Chang Qing , Djurdjanovic Dragan , Ni Jun , Lee Jay
Published: April 2007

Maintenance management has a direct influence on production performance. Existing works have not systematically taken the on-line production information into consideration in determining maintenance work-order priority, which is often assigned either through an ad hoc approach or using largely heuristic and static methods. In this paper, we first present a metric that can be used to quantitatively evaluate the effects of different maintenance priorities. Based on this index, one can employ a search algorithm to obtain maintenance work-order priorities that will lead to improved productivity within the optimization horizon. These concepts and methods are validated through simulation experiments and implementation in a real industrial facility. The results show that the effective utilization of on-line production data in dynamic maintenance scheduling can yield visible production benefit through maintenance priority optimization.
A Computational Response Surface Study of Three-Dimensional Aluminum Hemming Using Solid-to-Shell Mapping
Authors: Lin Guosong , Li Jing , Hu S. Jack , Cai Wayne
Published: April 2007

Hemming is a manufacturing process of folding a panel onto itself or another sheet. Quality of hemming is characterized by geometry and formability. This paper presents a response surface study of three-dimensional (3D) curved-surface-curved-edge hemming of an aluminum alloy, AA6111-T4, using finite-element (FE) analysis. Solid elements and explicit FE solver are used for simulations of flanging, pre- and final hemming, and shell elements with implicit solver are deployed for springback prediction. A novel procedure called “solid-to-shell mapping” is developed to bridge the solid elements with the shell elements. Verified to be accurate and efficient, the model is utilized in a central composite design to quantitatively explore the relationships between certain key process variables and the hem dimensional quality and formability. The most significant variables are identified as: (i) prehemming angle on roll-in/roll-out; (ii) nominal surface curvature on sheet springback; and (iii) initial sheet strain and flanging die radius on the maximum hemline surface strain of the produced hem. These results provide insights for process parameter selections in designing and optimizing 3D hems under material formability constraints.
Expanded Polystyrene Lost Foam Casting—Modeling Bead Steaming Operations
Authors: Matson Douglas M. , Venkatesh Rakesh , Biederman Scott
Published: April 2007

The retention of pattern voids observed in the production of expandable polystyrene patterns for lost foam casting can be traced to conditions developed during mold filling and subsequent steaming. Void formation and closure, or healing, was observed using high-speed video imaging through a clear acrylic sheet cut to match one-half of a test pattern mold. Two processing conditions, i.e., the initial bead packing density and the velocity of steam as it passes between beads, were shown to significantly impact the ability of a void to heal during steaming. A model is proposed to predict conditions where voids will heal based on three criteria that relate to a limitation of the processing window, the void size, and the ability of the bead to swell.
Experimental and Numerical Investigations of a Split-Ring Test for Springback
Authors: Xia Z. Cedric , Miller Craig E. , Ren Feng
Published: April 2007

This paper presents an in-depth experimental and numerical investigation of a split-ring test, which provides a simple yet effective benchmark for correlating forming and springback predictive capabilities with experimental measurements. The experimental procedure consists of deep drawing a circular 6111-T4 aluminum alloy into a cylindrical cup of 55mm depth, crosscutting nine rings each of 5mm wide from the cup, splitting the rings, and measuring their opening displacement, i.e., the springback amount. Experimental data obtained included punch force trajectories, drawn cup profile, thickness distribution after forming, and the ring openings after splitting. A numerical model is built to analyze the process, and both transversely isotropic and fully orthotropic yield criteria are investigated. Simulation results are validated against experimental data. A detailed numerical analysis is also conducted for stress distributions in each ring after each step and their relationship to the total springback amount. Stress and strain signatures suggested that the test is well suited for validating material models, such as anisotropic yield surface models and hardening models.
Automated Slicing for a Multiaxis Metal Deposition System
Authors: Ruan Jianzhong , Sparks Todd E. , Panackal Ajay , Liou F. W. , Eiamsa-ard Kunnayut , Slattery Kevin , Chou Hsin-Nan , Kinsella Mary
Published: April 2007

A multiaxis adaptive slicing algorithm for multiaxis layered manufacturing, which can generate optimal slices to achieve deposition without support structures, is presented in this paper. Different from current adaptive slicing, this technique varies not only layer thickness but also in slicing/building direction. Aware of potential problems of previous research on slicing, the work in this paper focuses on innovative geometry reasoning and analysis tool-centroidal axis. Similar to medial axis, it contains geometry and topological information but is significantly computationally cheaper. Using a centroidal axis as a guide, the multiaxis slicing procedure is able to generate a three-dimensional layer or change slicing direction as needed automatically to build the part with better surface quality. This paper presents various examples to demonstrate the feasibility and advantages of centroidal axis and its usage in the multiaxis slicing process.
Requirement of
bic/microRNA-155
for Normal Immune Function
Authors: Rodriguez Antony , Vigorito Elena , Clare Simon , Warren Madhuri V. , Couttet Philippe , Soond Dalya R. , van Dongen Stijn , Grocock Russell J. , Das Partha P. , Miska Eric A. , Vetrie David , Okkenhaug Klaus , Enright Anton J. , Dougan Gordon , Turner Martin , Bradley Allan
Published: April 2007

MicroRNAs are a class of small RNAs that are increasingly being recognized as important regulators of gene expression. Although hundreds of microRNAs are present in the mammalian genome, genetic studies addressing their physiological roles are at an early stage. We have shown that mice deficient for
bic/microRNA-155
are immunodeficient and display increased lung airway remodeling. We demonstrate a requirement of
bic/microRNA-155
for the function of B and T lymphocytes and dendritic cells. Transcriptome analysis of
bic/microRNA-155
–deficient CD4
+
T cells identified a wide spectrum of microRNA-155–regulated genes, including cytokines, chemokines, and transcription factors. Our work suggests that
bic/microRNA-155
plays a key role in the homeostasis and function of the immune system.
Immunization with PspA Incorporated into a Poly(Ethylene Oxide) Matrix Elicits Protective Immunity against
Streptococcus pneumoniae
Authors: Moore Quincy C. , Johnson LaShundra , Repka Michael , McDaniel Larry S.
Published: April 2007

CBA/N mice were immunized with PspA in a poly(ethylene oxide) matrix to examine its ability to deliver the antigen and modulate the immune response. All mice receiving PspA in the matrix survived a lethal pneumococcal challenge and had serum anti-PspA antibody levels statistically higher than mice receiving PspA alone (
P
< 0.009).
Extended Culture Enhances Sensitivity of a Gamma Interferon Assay for Latent
Mycobacterium tuberculosis
Infection
Authors: Cehovin Ana , Cliff Jacqueline M. , Hill Philip C. , Brookes Roger H. , Dockrell Hazel M.
Published: April 2007

To test the hypothesis that prolonged culture would enhance the sensitivity of latent tuberculosis detection by a gamma interferon release assay, blood samples from 33 household contacts of Gambian tuberculosis patients were stimulated with
Mycobacterium tuberculosis
-specific antigens. After 24 h of culture, 66% were positive, compared to 93% after 6 days of culture.
Seroprevalence of Selected Infectious Agents in a Free-Ranging, Low-Density Lion Population in the Central Kalahari Game Reserves in Botswana
Authors: Ramsauer Sandra , Bay Gert , Meli Marina , Hofmann-Lehmann Regina , Lutz Hans
Published: April 2007

Twenty-one free-ranging Central Kalahari lions (
Panthera leo
) exhibited a high prevalence rate of feline herpesvirus (100%) and feline immunodeficiency virus (71.4%). Canine distemper virus and feline calicivirus occurred with a low prevalence. All individuals tested negative for feline coronavirus, feline parvovirus, feline leukemia virus,
Ehrlichia canis
, and
Anaplasma phagocytophilum
.
Exocytosis of Neutrophil Formyl Peptide Receptor-Like 1 (fPRL1) Results in Downregulation of Cytoplasmic fPRL1 in Patients with Purulent Dermatitis
Authors: Ohara Eiji , Kumon Yoshitaka , Kobayashi Toshihiro , Takeuchi Hiroaki , Sugiura Tetsuro
Published: April 2007

N-Formyl peptide receptor-like 1 (fPRL1) is a member of the chemoattractant subfamily of G protein-coupled receptors and plays a key role in inflammation via chemotaxis and the regulation of mediator release from leukocytes. Activated fPRL1 has recently been shown to induce a complicated pattern of cellular signaling in vitro, but the details of the regulation and alteration of leukocyte cellular fPRL1 during inflammation in vivo remain unclear. To clarify the alteration of neutrophil fPRL1 during inflammation in vivo, the immunohistochemical staining of neutrophil fPRL1 in samples from patients with purulent dermatitis was performed. The in vitro morphological alteration of neutrophil fPRL1 on cellular membranes by stimulation withN-formylmethionyl-leucyl-phenylalanine (fMLP) was also examined. Both the cytoplasm and the cellular membranes of blood neutrophils stained strongly for fPRL1. On the other hand, the cellular membranes of neutrophils in dermatitis tissue stained strongly for fPRL1 but the cytoplasm stained weakly. The enhancement of neutrophil fPRL1 on cellular membranes by stimulation with fMLP indicates the exocytosis of neutrophil fPRL1-containing granules. In conclusion, we for the first time confirmed the alteration of neutrophil fPRL1 in clinical cases of purulent dermatitis. Cytoplasm that was weakly stained and cellular membranes that were well stained for fPRL1 were considered to be distinctive features of activated neutrophils in purulent dermatitis tissue.
Single-Chain Antibody Fragment Specific forPlasmodium vivaxDuffy Binding Protein
Authors: Kim So-Hee , Hwang Seung-Young , Lee Yong-Seok , Choi In-Hak , Park Sae-Gwang , Kho Weon-Gyu
Published: April 2007

Phage display of single-chain variable fragment (scFv) antibodies is a powerful tool for selecting important, useful, and specific human antibodies. We constructed a library from three patients infected withPlasmodium vivax. Panning on recombinant PvRII enriched a population of scFvs that recognized region II of theP. vivaxDuffy binding protein (DBP). Three clones of scFvs that reacted with PvRII were selected, and their biological functions were analyzed. These scFvs inhibited erythrocyte binding to DBP. Clone SFDBII92 had the greatest affinity (dissociation constant = 3.62 × 10−8M) and the greatest inhibition activity (50% inhibitory concentration ≈ 2.9 μg/ml) to DBP. Thus, we demonstrated that human neutralizing antibody could be made from malaria patients using phage display and that these neutralizing scFvs should prove valuable for developing both passive and active immunization strategies based on DBP.
Genetic and Phenotypic Features of Blood and Genital Viral Populations of Clinically Asymptomatic and Antiretroviral-Treatment-Naive Clade A Human Immunodeficiency Virus Type 1-Infected Women
Authors: Andreoletti Laurent , Skrabal Katharina , Perrin Virginie , Chomont Nicolas , Saragosti Sentob , Gresenguet Gerard , Moret Helene , Jacques Jerome , Longo Jean de Dieu , Matta Mathieu , Mammano Fabrizio , Belec Laurent
Published: April 2007

In the present study, we assessed whether human immunodeficiency virus type 1 (HIV-1) genetic compartmentalization was associated with phenotypic CCR5 (R5) or CXCR4 (X4) coreceptor usage differences between the systemic and the genital viral populations. Four clinically asymptomatic and treatment-naïve clade A HIV-1-infected patients were selected from a cohort of 274 African women, because they were free of all the biological cofactors known to modify the kinetics of viral production in the genital tract. HIV RNA envelope sequences (V1 to V3) derived from plasma and cervicovaginal secretions (CVS) were amplified, subcloned, and sequenced. CCR5 or CXCR4 coreceptor usage was determined by production of recombinant viral particles, followed by single-cycle infection assays of indicator cell lines, using the tropism recombinant test. In these four selected patients, CVS-derived sequences appeared to be genetically distinct from blood-derived sequences (
P
≤ 0.001). Two patients were found to harbor virus populations with only the R5 phenotype in both compartments, whereas viruses using CXCR4 in addition to CCR5 were detected in two other patients. In particular, one woman harbored genital virus populations with mixed R5 and X4 phenotypes associated with peripheral blood populations with only the R5 phenotype. These results demonstrate genetic compartmentalization of HIV between the plasma and genital secretions of clinically asymptomatic, treatment-naïve, clade A-infected women. Also, for one patient, we report phenotypic coreceptor usage differences between the systemic (R5) and genital (R5/X4) viral populations. These features may be critical for the development of further mucosal vaccines, therapies, or new preventive strategies to block heterosexual transmission.
Assessment of Fluorescent In Situ Hybridization and PCR-Based Methods for Rapid Identification of
Burkholderia cepacia
Complex Organisms Directly from Sputum Samples
Authors: Brown A. R. , Govan J. R. W.
Published: April 2007

Several species within the
Burkholderia cepacia
complex (BCC) have emerged as significant opportunistic pathogens of patients with cystic fibrosis (CF). BCC infection is typically associated with a poor clinical prognosis and decreased survival. These factors, combined with the existence of highly transmissible epidemic strains, have resulted in strict segregation of BCC- and non-BCC-infected patients to minimize cross infection. Accurate and rapid diagnosis of infections is essential to enable appropriate patient management. However, the rapidly evolving taxonomy of BCC poses a considerable challenge to diagnostics. In the present study, we assessed a commercially available fluorescent in situ hybridization (FISH) assay (seaFAST Cystic Fibrosis I kit) and a novel rRNA gene-based PCR assay for the rapid identification of BCC-positive sputa, irrespective of the BCC species. We report that, while the FISH assay fails to identify all BCC species, it does identify the majority of species, including the two most clinically relevant species,
B. multivorans
and
B. cenocepacia
. The sensitivity of the assay applied to sputum was limited by nonspecific background fluorescence. While sputum processing was optimized to minimize background, the resulting sensitivity for BCC detection was 8 × 10
5
CFU/ml. In contrast, the novel PCR assay reported herein exhibits 100% sensitivity and specificity for all BCC species and can detect 10
4
CFU/ml when applied to sputum. This novel rRNA gene-based assay is currently the most sensitive BCC-specific PCR assay for the detection of BCC direct from clinical samples and as such is a valuable addition to the field of BCC diagnostics.
Discovery of a Novel Human Picornavirus in a Stool Sample from a Pediatric Patient Presenting with Fever of Unknown Origin
Authors: Jones Morris S. , Lukashov Vladimir V. , Ganac Robert D. , Schnurr David P.
Published: April 2007

Fever of unknown origin (FUO) is a serious problem in the United States. An unidentified agent was cultured from the stool of an infant who presented with FUO. This virus showed growth in HFDK cells and suckling mice. Using DNase sequence-independent single-primer amplification, we identified several nucleotide sequences with a high homology to Theiler's murine encephalomyelitis virus. Nearly full-length viral genome sequencing and phylogenetic analysis demonstrate that this virus is a member of the
Cardiovirus
genus of the
Picornaviridae
family.
Nonoutbreak Surveillance of Group A Streptococci Causing Invasive Disease in Portugal Identified Internationally Disseminated Clones among Members of a Genetically Heterogeneous Population
Authors: Friães A. , Ramirez M. , Melo-Cristino J.
Published: April 2007

The typing of 160 invasive
Streptococcus pyogenes
isolates confirmed the importance of pulsed-field gel electrophoresis and multilocus sequence typing for defining clones. The results identified an extremely diverse population and highlighted the importance of both internationally disseminated and local clones not previously associated with invasive disease.
Nucleotide Sequence Diversity of the
bexA
Gene in Serotypeable
Haemophilus influenzae
Strains Recovered from Invasive Disease Patients in Canada
Authors: Zhou Jianwei , Law Dennis K. S. , Sill Michelle L. , Tsang Raymond S. W.
Published: April 2007

The
bexA
genes of 36
Haemophilus influenzae
isolates were sequenced to reveal their nucleotide sequence diversity, which divided them into two groups, similar to clonal divisions I and II. This sequence diversity may lead to false-negative PCR results for
H. influenzae
infections if
bexA
is the chosen gene target.
Adenovirus E4 34k and E1b 55k Oncoproteins Target Host DNA Ligase IV for Proteasomal Degradation
Authors: Baker Amy , Rohleder Kent J. , Hanakahi Les A. , Ketner Gary
Published: April 2007

Cells infected by adenovirus E4 mutants accumulate end-to-end concatemers of the viral genome that are assembled from unit-length viral DNAs by nonhomologous end joining (NHEJ). Genome concatenation can be prevented by expression either of E4 11k (product of E4orf3) or of the complex of E4 34k (product of E4orf6) and E1b 55k. Both E4 11k and the E4 34k/E1b 55k complex prevent concatenation at least in part by inactivation of the host protein Mre11: E4 11k sequesters Mre11 in aggresomes, while the E4 34k/E1b 55k complex participates in a virus-specific E3 ubiquitin ligase that mediates ubiquitination and proteasomal degradation. The E4 34k/E1b 55k complex, but not E4 11k, also inhibits NHEJ activity on internal breaks in the viral genome and on V(D)J recombination substrate plasmids, suggesting that it may interfere with NHEJ independently of its effect on Mre11. We show here that DNA ligase IV, which performs the joining step of NHEJ, is degraded as a consequence of adenovirus infection. Degradation is dependent upon E4 34k and E1b 55k, functional proteasomes, and the activity of cellular cullin 5, a component of the adenoviral ubiquitin ligase. DNA ligase IV also interacts physically with E1b 55k. The data demonstrate that DNA ligase IV, like Mre11, is a substrate for the adenovirus-specific E3 ubiquitin ligase; identify an additional viral approach to prevention of genome concatenation; and provide a mechanism for the general inhibition of NHEJ by adenoviruses.
Establishment of a Cottontail Rabbit Papillomavirus/HLA-A2.1 Transgenic Rabbit Model
Authors: Hu Jiafen , Peng Xuwen , Budgeon Lynn R. , Cladel Nancy M. , Balogh Karla K. , Christensen Neil D.
Published: April 2007

Three transgenic rabbit lines that express a well-characterized human major histocompatibility complex class I (MHC-I) gene (HLA-A2.1) have been established. All three lines carry the HLA-A2.1 heavy chain and are able to pass the transgene to their offspring with both the outbred and the inbred EIII/JC genetic background. HLA-A2.1 colocalizes exclusively with rabbit MHC-I on the cell surfaces. These HLA-A2.1 transgenic rabbits demonstrated infection patterns similar to those found after cottontail rabbit papillomavirus (CRPV) challenge when compared with results in normal rabbits, although higher regression rates were found in HLA-A2.1 transgenic rabbits. Because the CRPV genome can accommodate significant modifications, the CRPV/HLA-A2.1 rabbit model has the potential to be used to screen HLA-A2.1-restricted immunogenic epitopes from human papillomaviruses in the context of in vivo papillomavirus infection.
Atypical bZIP Domain of Viral Transcription Factor Contributes to Stability of Dimer Formation and Transcriptional Function
Authors: Schelcher Celine , Al Mehairi Salama , Verrall Elizabeth , Hope Questa , Flower Kirsty , Bromley Beth , Woolfson Derek N. , West Michelle J. , Sinclair Alison J.
Published: April 2007

The Epstein-Barr virus transcription factor Zta (encoded by
BZLF1
) is a bZIP protein containing an α-helical coiled-coil homodimerization motif (zipper). The Zta zipper forms less-stable dimers than other bZIP proteins, and an adjacent region (CT) interacts with the zipper to form a novel structure that is proposed to strengthen the dimer. Here we question the role of the CT region for Zta function. Cross-linking experiments demonstrate that the entire CT region lies adjacent to the zipper. Detailed analyses of Zta truncation mutations identify an involvement of the proximal CT region (221 to 230) in dimer formation with a further contribution from the distal region (236 to 243). Biophysical analyses reveal that residues 221 to 230 enhance the stability of the coiled coil. The ability of the Zta truncation mutants to interact with three Zta-binding sites also requires the proximal CT region. Fine mapping of DNA-binding requirements highlighted the contribution of these amino acids for Zta function. Thus, the proximal part of the CT region is required to aid the dimerization of Zta and thereby its DNA-binding ability. In contrast, although the distal part of the CT region aids dimerization, it promotes only a modest increase in DNA binding. To probe this further, we defined the contribution from the CT region for Zta to transactivate a promoter embedded within the viral genome. From this we conclude that the proximal part of the CT region is absolutely required, whereas the distal part is dispensable.
Detection of a Novel and Highly Divergent Coronavirus from Asian Leopard Cats and Chinese Ferret Badgers in Southern China
Authors: Dong B. Q. , Liu W. , Fan X. H. , Vijaykrishna D. , Tang X. C. , Gao F. , Li L. F. , Li G. J. , Zhang J. X. , Yang L. Q. , Poon L. L. M. , Zhang S. Y. , Peiris J. S. M. , Smith G. J. D. , Chen H. , Guan Y.
Published: April 2007

Since an outbreak of severe acute respiratory syndrome (SARS) was averted in 2004, many novel coronaviruses have been recognized from different species, including humans. Bats have provided the most diverse assemblages of coronaviruses, suggesting that they may be the natural reservoir. Continued virological surveillance has proven to be the best way to avert this infectious disease at the source. Here we provide the first description of a previously unidentified coronavirus lineage detected from wild Asian leopard cats (
Prionailurus bengalensis
) and Chinese ferret badgers (
Melogale moschata
) during virological surveillance in southern China. Partial genome analysis revealed a typical coronavirus genome but with a unique putative accessory gene organization. Phylogenetic analyses revealed that the envelope, membrane, and nucleoprotein structural proteins and the two conserved replicase domains, putative RNA-dependent RNA polymerase and RNA helicase, of these novel coronaviruses were most closely related to those of group 3 coronaviruses identified from birds, while the spike protein gene was most closely related to that of group 1 coronaviruses from mammals. However, these viruses always fell into an outgroup phylogenetic relationship with respect to other coronaviruses and had low amino acid similarity to all known coronavirus groups, indicating that they diverged early in the evolutionary history of coronaviruses. These results suggest that these viruses may represent a previously unrecognized evolutionary pathway, or possibly an unidentified coronavirus group. This study demonstrates the importance of systematic virological surveillance in market animals for understanding the evolution and emergence of viruses with infectious potential.
Acute Resolving Woodchuck Hepatitis Virus (WHV) Infection Is Associated with a Strong Cytotoxic T-Lymphocyte Response to a Single WHV Core Peptide
Authors: Frank Ina , Budde Claudia , Fiedler Melanie , Dahmen Uta , Viazov Sergei , Lu Mengji , Dittmer Ulf , Roggendorf Michael
Published: April 2007

Woodchucks infected with woodchuck hepatitis virus (WHV) are an excellent model for studying acute, self-limited and chronic hepadnaviral infections. Defects in the immunological response leading to chronicity are still unknown. Specific T-helper cell responses to WHV core and surface antigens (WHcAg and WHsAg, respectively) are associated with acute resolving infection; however, they are undetectable in chronic infection. Up to now, cytotoxic T-lymphocyte (CTL) responses could not be determined in the woodchuck. In the present study, we detected virus-specific CTL responses by a CD107a degranulation assay. The splenocytes of woodchucks in the postacute phase of WHV infection (18 months postinfection) were isolated and stimulated with overlapping peptides covering the whole WHcAg. After 6 days, the cells were restimulated and stained for CD3 and CD107a. One peptide (c96-110) turned out to be accountable for T-cell expansion and CD107a staining. Later, we applied the optimized degranulation assay to study the kinetics of the T-cell response in acute WHV infection. We found a vigorous T-cell response against peptide c96-110 with peripheral blood cells beginning at the peak of viral load (week 5) and lasting up to 15 weeks postinfection. In contrast, there was no T-cell response against peptide c96-110 detectable in chronically WHV-infected animals. Thus, with this newly established flow cytometric degranulation assay, we detected for the first time virus-specific CTLs and determined one immunodominant epitope of WHcAg in the woodchuck.
Identification of Residues Critical for the Interferon Antagonist Function of Langat Virus NS5 Reveals a Role for the RNA-Dependent RNA Polymerase Domain
Authors: Park Gregory S. , Morris Keely L. , Hallett Roselyn G. , Bloom Marshall E. , Best Sonja M.
Published: April 2007

All pathogenic flaviviruses examined thus far inhibit host interferon (IFN) responses by suppressing the Janus kinase-signal transducer and activator of transcription (JAK-STAT) pathway. Both Langat virus (LGTV; a member of the tick-borne encephalitis virus serogroup) and Japanese encephalitis virus use the nonstructural protein NS5 to suppress JAK-STAT signaling. However, NS5 is also critical to virus replication, contributing methyltransferase and RNA-dependent RNA polymerase (RdRP) activities. The specific amino acid residues of NS5 involved in IFN antagonism are not known. Here, we demonstrate that the LGTV NS5 JAK-STAT inhibitory domain is contained between amino acids 355 and 735 (of 903), a range which lies within the RdRP domain. Furthermore, we identified two noncontiguous stretches of specific amino acids within the RdRP, 374 to 380 and 624 to 647, as critical for inhibition of JAK-STAT signaling. Despite considerable separation on the linear NS5 sequence, these residues localized adjacent to each other when modeled on the West Nile virus RdRP crystal structure. Due to the general conservation of RdRP structures, these results suggest that the specific residues identified act cooperatively to form a unique functional site on the RdRP responsible for JAK-STAT inhibition. This insight into the mechanism underlying flavivirus IFN evasion strategies will facilitate the design of antiviral therapeutics that potentiate the action of IFN during infection.
SOME RELATIONS AMONG VALUES, INTERACTIONS, AND DECOMPOSABILITY OF NON-ADDITIVE MEASURES
Authors: FUJIMOTO KATSUSHIGE , MUROFUSHI TOSHIAKI
Published: April 2007

We will discuss some relations among values, i.e., global / overall importance of each single element of the object to be considered, interaction indices, i.e., simultaneous interaction among the elements in a coalition, and inclusion-exclusion coverings, i.e., decomposability of non-additive measures and / or of the Choquet integral. So, we show that inclusion-exclusion coverings are characterized by the Shapley interaction index. Then, inclusion-exclusion coverings are invariant under the transformations which characterize the Shapley value and / or the Shapley interaction index, and preserve the Shapley value.
Relationship between Kir2.1/Kir2.3 activity and their distributions between cholesterol-rich and cholesterol-poor membrane domains
Authors: Tikku Saloni , Epshtein Yulia , Collins Heidi , Travis Alexander J. , Rothblat George H. , Levitan Irena
Published: April 2007

Our earlier studies have shown that Kir2.x channels are suppressed by an increase in the level of cellular cholesterol, whereas cholesterol depletion enhances the activity of the channels. In this study, we show that Kir2.1 and Kir2.3 channels have double-peak distributions between cholesterol-rich (raft) and cholesterol-poor (non-raft) membrane fractions, indicating that the channels exist in two different types of lipid environment. We also show that whereas methyl-β-cyclodextrin-induced cholesterol depletion removes cholesterol from both raft and non-raft membrane fractions, cholesterol enrichment results in cholesterol increase exclusively in the raft fractions. Kinetics of both depletion-induced Kir2.1 enhancement and enrichment-induced Kir2.1 suppression correlate with the changes in the level of raft cholesterol. Furthermore, we show not only that cholesterol depletion shifts the distribution of the channels from cholesterol-rich to cholesterol-poor membrane fractions but also that cholesterol enrichment has the opposite effect. These observations suggest that change in the level of raft cholesterol alone is sufficient to suppress Kir2 activity and to facilitate partitioning of the channels to cholesterol-rich domains. Therefore, we suggest that partitioning to membrane rafts plays an important role in the sensitivity of Kir2 channels to cholesterol.
Thigh muscle activation distribution and pulmonary V̇o2kinetics during moderate, heavy, and very heavy intensity cycling exercise in humans
Authors: Endo Masako Yamaoka , Kobayakawa Mayumi , Kinugasa Ryuta , Kuno Shinya , Akima Hiroshi , Rossiter Harry B. , Miura Akira , Fukuba Yoshiyuki
Published: April 2007

The mechanisms underlying the oxygen uptake (V̇o2) slow component during supra-lactate threshold (supra-LT) exercise are poorly understood. Evidence suggests that the V̇o2slow component may be caused by progressive muscle recruitment during exercise. We therefore examined whether leg muscle activation patterns [from the transverse relaxation time (T2) of magnetic resonance images] were associated with supra-LT V̇o2kinetic parameters. Eleven subjects performed 6-min cycle ergometry at moderate (80% LT), heavy (70% between LT and critical power; CP), and very heavy (7% above CP) intensities with breath-by-breath pulmonary V̇o2measurement. T2 in 10 leg muscles was evaluated at rest and after 3 and 6 min of exercise. During moderate exercise, nine muscles achieved a steady-state T2 by 3 min; only in the vastus medialis did T2 increase further after 6 min. During heavy exercise, T2 in the entire vastus group increased between minutes 3 and 6, and additional increases in T2 were seen in adductor magnus and gracilis during this period of very heavy exercise. The V̇o2slow component increased with increasing exercise intensity (being functionally zero during moderate exercise). The distribution of T2 was more diverse as supra-LT exercise progressed: T2 variance (ms) increased from 3.6 ± 0.2 to 6.5 ± 1.7 between 3 and 6 min of heavy exercise and from 5.5 ± 0.8 to 12.3 ± 5.4 in very heavy exercise (rest = 3.1 ± 0.6). The T2 distribution was significantly correlated with the magnitude of the V̇o2slow component ( P < 0.05). These data are consistent with the notion that the V̇o2slow component is an expression of progressive muscle recruitment during supra-LT exercise.
Central and baroreflex control of heart period during the wake-sleep cycle in spontaneously hypertensive rats
Authors: Berteotti Chiara , Asti Valentina , Ferrari Vera , Franzini Carlo , Lenzi Pierluigi , Zoccoli Giovanna , Silvani Alessandro
Published: April 2007

We investigated whether the relative contribution of the baroreflex and central commands to the control of heart period differs between spontaneously hypertensive rats (SHR) and Wistar-Kyoto normotensive rats (WKY) during physiological behavior. Rats were instrumented with an arterial catheter and with electrodes for discriminating wakefulness, nonrapid eye movement sleep (NREMS), and rapid eye movement sleep (REMS). The cross-correlation function (CCF) between spontaneous fluctuations of heart period and mean arterial pressure was computed at frequencies <0.2 Hz. The baroreflex determines a positive correlation between heart period and previous pressure values. This pattern was observed in the CCF during quiet wakefulness (QW) and NREMS, and in QW, it was accompanied by a pronounced negative correlation between heart period and subsequent pressure values. The relative baroreflex contribution to the control of heart period, estimated from the positive peak value of the CCF, was lower in SHR than in WKY during QW but not during NREMS. During REMS, the CCF showed a negative correlation between heart period and both previous and subsequent pressure values, reflecting the prevalence of central autonomic commands. The relative contribution of central commands to the control of heart period, estimated from the negative peak value of the CCF, was lower in SHR than in WKY during REMS. These results suggest that during QW and REMS, the control of heart period exerted by the baroreflex and central commands, respectively, is less effective in SHR than in WKY. This difference is not apparent in a behavioral state of autonomic stability such as NREMS.