bluuebunny/crossref_metadata_2025 huggingface dataset:
Do female spotless starlings Sturnus unicolor adjust maternal investment according to male attractiveness?
Authors: López‐Rull Isabel , Gil Diego
Published: March 2009

In birds, female egg allocation patterns have a strong influence in offspring development and differential investment in egg size and composition has been shown to respond to male attractiveness. In this study we experimentally manipulated the perceived attractiveness of male starlings Sturnus unicolor by increasing the amount of green material in some nests (a male courtship display in this species). We predicted that, if female investment before laying is related to male attractiveness, experimental females would increase their reproductive investment in response to the addition of plants in their nests when compared to control females. We found that our manipulation caused variations in female reproductive investment in a way that seems to influence offspring quantity but not offspring quality: Females laid larger clutch sizes but not larger eggs when green plant material was added. However, yolk androgens contents were not related to the experimental manipulation. Contrary to expectations, females breeding in experimental nests laid eggs with smaller amounts of eggshell pigments. Interestingly, we found that eggs laid later in the sequence had higher testosterone levels and showed more intense egg colouration than eggs laid earlier in the sequence. These differences at the within‐clutch level suggest that selection has favoured compensatory strategies for hatching asynchrony. Alternatively, since nest sabotages by other females are most common at the beginning of laying, this could be seen as female strategy to minimise losses due to nest sabotages. As far as we know, this is the first study to show that an external egg characteristic such as blue‐green colouration reflects yolk androgen concentration.
Enamel defects and dental caries in 9‐year‐old children living in fluoridated and nonfluoridated areas of Auckland, New Zealand
Authors: Kanagaratnam Sathananthan , Schluter Philip , Durward Callum , Mahood Robyn , Mackay Tim
Published: March 2009

Abstract –  Objectives:  This epidemiological study aims to investigate the developmental enamel defects and dental caries among 9‐year‐old children resident in fluoridated and nonfluoridated regions in Auckland, New Zealand.

Methods:  A stratified, two‐stage random selection design where strata were defined by fluoridation status, school size, and school decile. After informed consent was obtained, parents completed oral health questionnaires and children underwent dental examinations at school clinics.

Results:  612 children from 38 schools participated in the study. Overall, 175 (29%) children had lived continuously in fluoridated areas, 149 (24%) had lived continuously in nonfluoridated areas, and 288 (47%) had resided intermittently in fluoridated areas. Diffuse opacities were present in 117 (19%) children and deciduous teeth dental caries was seen in 370 (60%) children. After adjustment for covariates, a strong dose–response relationship between diffuse opacity and fluoridation status was found, with children who lived continuously in fluoridated areas being 4.17 times as likely to have diffuse opacities as children who lived continuously in nonfluoridated areas (P < 0.001). Conversely, a strong protective dose–response relationship between caries experience and fluoridation status was seen, with children who lived continuously in fluoridated areas being 0.42 times as likely to have dental caries as children who lived continuously in nonfluoridated areas (P < 0.001).

Conclusions:  Reticulated water fluoridation in Auckland reduces the risk of dental caries but increases the risk of diffuse opacities in 9‐year‐old children. Guidelines and health‐promotion strategies that enable children to minimize their risk to diffuse opacities yet reduce their risk of dental caries should be reviewed.
Risk factors for severe infection in patients with hairy cell leukemia: a long‐term study of 73 patients
Authors: Damaj Ghandi , Kuhnowski Frédérique , Marolleau Jean‐Pierre , Bauters Francis , Leleu Xavier , Yakoub‐Agha Ibrahim
Published: March 2009

Although the survival of patients with hairy cell leukemia (HCL) has been improved by the therapeutic introduction of interferon α and purine analogs, it is still worsened by complications such as severe infections. In this long‐term study, we identified factors influencing patient outcomes in 73 patients with HCL. Median age at diagnosis was 53 yr and the gender ratio (M/F) was 2.3. At the time of HCL diagnosis, 60 patients (82%) were symptomatic and 22 of these had an infection. After a median follow‐up of 13 yr, eight patients had died of secondary cancer (n = 2), HCL progression (n = 1) and age‐related complications (n = 5). The 10‐yr overall survival (OS), progression‐free survival and relapse rates were 91 ± 3%, 14 ± 5% and 87 ± 5%, respectively. In multivariate analyses, age >53 yr was the only factor adversely influencing OS and secondary cancer incidence, with adjusted hazard ratio (HR) of 9.30 (95%CI, 1.15–76.6; P = 0.037) and 2.80 (95%CI, 1.05–7.71; P = 0.04), respectively. Eleven patients developed severe infections. Absolute lymphocyte count (<1 × 109/L) at diagnosis was the only factor influencing the occurrence of severe infections, with an adjusted HR of 4.01 (P = 0.007). Strikingly, we did not observe any significant correlation between neutrophil or monocyte counts and the incidence of infection. We confirmed long‐term survival in HCL but found a high incidence of infection – even late in the course of the disease. The absolute lymphocyte count at diagnosis is a risk factor for the occurrence of severe infections. In addition to careful monitoring of infections, prompt initiation of anti‐HCL treatment should be considered in patients with low lymphocyte counts.
The induction of tumor‐specific CD4+ T cells via major histocompatibility complex class II is required to gain optimal anti‐tumor immunity against B16 melanoma cell line in tumor immunotherapy using dendritic cells
Authors: Fujisawa Yasuhiro , Nabekura Tsukasa , Nakao Tomohei , Nakamura Yasuhiro , Takahashi Takenori , Kawachi Yasuhiro , Otsuka Fujio , Onodera Masafumi
Published: March 2009

Abstract:  We have demonstrated that dendritic cells (DCs) genetically modified to express tumor‐associated antigens (TAAs) with retroviral vectors elicit more potential anti‐tumor effect than those loaded with peptides because they can prime antigen‐specific CD4+ T cells resulting in production of tumor‐specific antibody. In this study, we showed the importance of antigen presentation via a major histocompatibility complex (MHC) class II molecule in cancer immunity against non‐membrane bound TAAs such as the melanoma antigen gp100 by using DCs derived from MHC class II‐deficient mice (C2KO). DCs were prepared by transduction of gp100 cDNA into haematopoietic progenitor cells obtained from C2KO followed by differentiation with cytokines (C2KO‐gp/DCs). When C2KO‐gp/DCs were inoculated into immunocompetent mice, the mice scarcely primed the antigen‐specific Th1 cells and developed fewer CD8 T cells than did those inoculated with transduced DCs prepared from normal mice. The attenuated anti‐tumor effect was also confirmed in a postimmunization setting where, while two of eight control mice eradicated the pre‐existing melanoma cell line B16 (25%), no mice inoculated with C2KO‐gp/DCs did. These results suggested not only the limitation of current protocols using MHC class I‐restricted tumor peptides but also the usefulness of DCs expressing gp100 in vaccine therapy against melanoma.
Lipid‐Induced ER Stress: Synergistic Effects of Sterols and Saturated Fatty Acids
Authors: Pineau Ludovic , Colas Jenny , Dupont Sébastien , Beney Laurent , Fleurat‐Lessard Pierrette , Berjeaud Jean‐Marc , Bergès Thierry , Ferreira Thierry
Published: March 2009

Stress within the endoplasmic reticulum (ER) induces a coordinated response, namely the unfolded protein response (UPR), devoted to helping the ER cope with the accumulation of misfolded proteins. Failure of the UPR plays an important role in several human diseases. Recent studies report that intracellular accumulation of saturated fatty acids (SFAs) and cholesterol, seen in diseases of high incidence, such as obesity or atherosclerosis, results in ER stress. In the present study, we evaluated the effects of perturbations to lipid homeostasis on ER stress/UPR induction in the model eukaryote Saccharomyces cerevisiae. We show that SFA originating from either endogenous (preclusion of fatty acid desaturation) or exogenous (feeding with extracellular SFA) sources trigger ER stress and that ergosterol, the major sterol in yeast, acts synergistically with SFA in this process. This latter effect is connected to ergosterol accumulation within microsomal fractions from SFA‐accumulating cells, which display highly saturated phospholipid content. Moreover, treating the cells with the molecular chaperone 4‐phenyl butyrate abolishes UPR induction, suggesting that lipid‐induced ER stress leads to an overload of misfolded protein that acts, in turn, as the molecular signal for induction of the UPR. The present data are discussed in the context of human diseases that involve lipid deregulation.
Environmental tobacco smoke exposure as a risk factor for infections in infancy
Authors: Ladomenou F , Kafatos A , Galanakis E
Published: March 2009

Aim:  This prospective observational study investigated the effect of environmental tobacco smoke (ETS) on frequency and severity of common infantile infections.

Methods:  In a representative sample of 926 infants, parental smoking was recorded at months 1 and 9 postpartum, and all infantile infectious episodes were recorded at 1, 3, 6, 9 and 12 months postpartum.

Results:  Both parents were regular smokers all through the first year in 107 (11.6%), at least one smoked regularly or occasionally in 492 (53.1%), and parents did not smoke at all in 327 (35.3%) families. Among mothers, 168 (18.1%) smoked perinatally. Infantile ETS exposure was associated with increased frequency of total infectious episodes (p = 0.025) and hospitalizations for infection (p = 0.007). In ETS exposed infants, birth in autumn and presence of siblings contributed to increased frequency of most infections and of hospital admissions for infection. By contrast, exclusive breastfeeding protected against the effect of ETS on total infantile infections (OR 0.982, 95% CI 0.965–0.999; p = 0.036), hospital admissions for infection (OR 0.980, 95% CI 0.961–0.999; p = 0.036) and thrush (OR 0.973, 95% CI 0.951–0.996; p = 0.022).

Conclusion:  Our findings point to harmful effect of ETS on infantile health and further suggest that this effect may be enhanced or diminished by other factors. ETS should be regarded as a preventable risk factor for infections in infancy.
The Segmented Trend Line of Highest Life Expectancies
Authors: Vallin Jacques , Meslé France
Published: March 2009

The well‐known Oeppen–Vaupel straight line of maximum female life expectancies showed that the highest life expectancy observed in a given year increased linearly from 1840 to 2000. Their analysis fueled major controversy, especially when used to extrapolate future improvements in life expectancy at the same pace. We improve on the empirical analysis by enriching the dataset, expanding the period to 1750–2005, and considering both maximum life expectancy at birth and lowest age‐specific survival rates. It clearly appears that the original Oeppen–Vaupel straight line must be divided into several segments characterized by different slopes and that each segment corresponds to a major advance in the health transition. There is room to push life expectancy higher, but unless some new breakthrough increases the human life span, progress will very likely decelerate as mortality reduction affects individuals at older and older ages. The main key to the future lies not in knowing whether the observed straight line can be extrapolated but in anticipating the next major health improvement that will lead to an additional increase in life expectancy.
Hypertension in Asian/Pacific Island Americans
Authors: Watson Ralph E. , Karnchanasorn Rudruidee , Gossain Ved V.
Published: March 2009

Asian/Pacific Islander Americans (APIAs) are the fastest growing population in the United States by percentage. Hypertension is common and increases cardiovascular risk to a great extent in this population. The medical problems of this group are being increasingly encountered by US physicians. Many gene mutations associated with hypertension are more common in Asians. The significance of these polymorphisms in the pathogenesis of hypertension in APIAs is unclear. The percentage of APIAs who are aware, treated, and controlled is small. There may be some differences in the responses to antihypertensive medications between APIAs and whites. The results of human studies on the effect of drinking of tea on blood pressure in different groups are conflicting. Cough associated with angiotensin‐converting enzyme inhibitor therapy may be more common in APIAs than in whites. There is a need for more education of APIAs regarding hypertension and for more effective treatment of hypertension by the physicians caring for this population.
The Impact of Short‐Term Study Abroad Programs on L2 Listening Comprehension Skills
Authors: Cubillos Jorge H. , Chieffo Lisa , Fan Chunbo
Published: March 2009

Abstract: This study investigates the impact of a five‐week intermediate Spanish course on the listening comprehension ability of 48 participants in short‐term study abroad programs, and compares these groups with 92 peers enrolled in a similar course on the home campus. While both on‐campus and study abroad groups experienced similar gains in listening comprehension, there were significant differences in the way learners approached listening tasks: The study abroad group applied primarily top‐down and social listening strategies, while the on‐campus students favored bottom‐up processing. Higher‐proficiency students in the study abroad groups had significantly higher comprehension gains, and the study abroad groups achieved higher levels of confidence and self‐perceived ability after the treatment. Results outline some of the benefits and limitations of short‐term foreign sojourns for beginner and low‐intermediate language learners.
Foreign Language Learning and SAT Verbal Scores Revisited
Authors: Cooper Thomas C. , II Daniel J. Yanosky , Wisenbaker Joseph M. , Jahner David , Webb Elizabeth , Wilbur Marcia L.
Published: March 2009

Abstract: The purpose of the study was to investigate the relationship between foreign language learning and verbal ability in English as measured by the verbal section of the Scholastic Aptitude Test (SAT) Reasoning Test. Comparing foreign language students to nonforeign language students in this study, the effect of taking a foreign language on SAT verbal performance differed depending on how a student scored on the Preliminary SAT (PSAT) verbal subtest. Students with lower PSAT verbal scores had the greatest benefit from taking a foreign language, while students with higher PSAT verbal scores had smaller benefits. At the same time, within the range of possible PSAT verbal scores, students who took a foreign language outperformed students who did not. When the focus was on the language taken and the highest level attained by the end of junior year, the main effects for both foreign language taken and highest level were statistically significant though their partial eta squared measures suggested they had relatively low effect sizes. Pairwise comparisons indicated that outcomes for students taking Spanish were somewhat less than those for students taking Latin and German, and that students who took level III of their foreign language by the end of their junior year outperformed those who reached only levels I or 11.
Why Learning To Write Chinese Is a Waste of Time: A Modest Proposal
Authors: Allen Joseph R.
Published: March 2009

Abstract: This article argues that for students of Chinese and Japanese, learning to write Chinese characters (hanzi/kanji) by hand from memory is an inefficient use of resources. Rather, beginning students should focus on character/word recognition (reading) and electronic writing. Although electronic technologies have diminished the usefulness of Chinese character handwriting, its cultural importance remains. This leads to a hegemony of hanzi/kanji through which the assumed primacy of the written language is reinforced. After reviewing these conditions, strategies are offered to integrate handwriting skills with the new electronic writing technologies, creating an efficient and culturally sensitive program of instruction in hanzi/kanji writing. The article concludes with suggestions for further research needed to explore the theses of the essay.
The Overwhelmed Generation and Foreign Language Teacher Preparation: Developing Strategies To Work With the Mental Health Challenges of Preservice Teachers
Authors: Schrier Leslie L.
Published: March 2009

Abstract:For the last two decades the foreign language education profession has spent considerable energy establishing professional guidelines for foreign language teacher preparation. This article discusses research that suggests that the profession should now direct its energies toward developing protocols to assist preservice teachers whose mental health challenges place them at risk in their quest to become foreign language teachers. Further, the article discusses why teacher educators are ill prepared to cope with the substantial mental health issues undergraduates currently face. Examples are presented from a teacher education program that modified its supervision process to facilitate the development of strategies for coping with the stressors related to foreign language teaching in particular. Suggestions are made for research and policy changes to meet the mental health needs of current and future novice foreign language teachers.
Podcasting Communities and Second Language Pronunciation
Authors: Lord Gillian
Published: March 2009

Abstract: Although often neglected in language classrooms, second language phonology is a crucial element in language learning because it is often the most salient feature in the speech of a foreigner. As instructors, we must decide how to emphasize pronunciation and what techniques to use. This article discusses a collaborative pod‐casting project in an undergraduate Spanish phonetics class. Students worked in small groups to create and maintain their own podcast channel on which they uploaded recordings for group member feedback. Each recording focused on particular aspects of Spanish pronunciation, using tongue twisters, short readings, and personal reflection on students' own pronunciation. Both attitudes and pronunciation abilities were assessed before and after the project, and both were found to improve. The benefits of podcasting projects to improve language skills are discussed from an empirical as well as a pedagogical perspective.
The Interactive Effects of Pragmatic‐Eliciting Tasks and Pragmatic Instruction
Authors: Fukuya Yoshinori J. , Martínez‐Flor Alicia
Published: March 2009

Abstract:the effects of data‐gathering methods on pragmatic data have been well documented, yet an inquiry into the interactive effects of assessment tasks with pragmatic instruction has received scant attention. This study investigated the interaction between two assessment tasks (e‐mail and phone) and two types of pragmatic instruction (explicit and implicit). Forty‐nine Spanish learners of English engaged in these two tasks as pre‐ and posttests. The explicit group received 12 hours of metapragmatic information on head acts and hedges in suggestions while the implicit group was the recipient of recast and input enhancement activities. The results showed that postin‐structional improvement of the explicit condition was significantly more than that of the implicit condition in the phone task, although improvements of these two conditions were on par in the e‐mail task. This task‐induced variability might have been caused by an interaction between the feature of the two types of knowledge (i.e., monitoring capability) and an access to the knowledge bases (i.e., the role of attention to appropriateness and accuracy) in the two tasks.
An Analysis of Word Decision Strategies among Learners of Chinese
Authors: Shen Helen H.
Published: March 2009

Abstract: This study examines word decision strategies that 40 U.S. college beginning and advanced learners oj Chinese applied during reading. It first compares similarities and differences between the two groups in choosing strategies when making word decisions, then identifies effective word decision strategies from more than a hundred strategies used by learners. Results of the study indicate that although learners use various strategies requiring multiple levels of linguistic knowledge, the strategy of matching the target item to the learners' existing mental lexicon is predominant for both beginning and advanced learners. The results also show that the strategy of using contextual information to make word decisions is more frequently employed at the advanced level. In addition, the study reveals that the strategies required to use multiple linguistic cues and context information are better predictors of accurate word decision than other strategies.
ESL Theory‐Practice Dynamics: The Difficulty of Integrating Sociocultural Perspectives into Pedagogical Practices
Authors: Ajayi Lasisi
Published: March 2009

Abstract:The purpose of this study is to examine English as a second language (ESL) teachers' sensitivity to their students' social‐cultural backgrounds and the kind of constraints they face as they translate sociocultural perspectives into practice. Through both quantitative (questionnaire surveys) and qualitative (interviews) analysis, the study reports how 73 middle and high school teachers in Los Angeles related their own pedagogical practices to the social‐cultural experiences of their students. This researcher analyzed the quantitative data using descriptive statistics while qualitative data analysis was carried out using the framework of critical rejlexivity. Results showed that the teachers in the study were aware of and sensitive to the crucial role of their students' background experiences. However, the teachers pointed to institutional barriers and other constraints that have practical impacts on their sociocultural practices and outcomes.
The role of mammalian cardiac t‐tubules in excitation–contraction coupling: experimental and computational approaches
Authors: Orchard Clive H. , Pásek Michal , Brette Fabien
Published: March 2009

The sarcolemmal membrane of mammalian cardiac ventricular myocytes is characterized by the presence of invaginations called transverse tubules (t‐tubules). Transverse tubules occur at the Z‐line as transverse elements with longitudinal extensions. While the existence of t‐tubules has been known for some time, recent experimental studies have suggested that their structure and function are more complex than previously believed. There are, however, aspects of t‐tubule function that are not currently amenable to experimental investigation, but can be investigated using computational and mathematical approaches. Such studies have helped elucidate further the possible role of t‐tubules in cell function. This review summarizes recent experimental and complementary computational studies which highlight the important role of t‐tubules in cardiac excitation–contraction coupling.
Variants of the Antibody Herceptin That Interact with HER2 and VEGF at the Antigen Binding Site
Authors: Bostrom Jenny , Yu Shang-Fan , Kan David , Appleton Brent A. , Lee Chingwei V. , Billeci Karen , Man Wenyan , Peale Franklin , Ross Sarajane , Wiesmann Christian , Fuh Germaine
Published: March 2009

The interface between antibody and antigen is often depicted as a lock and key, suggesting that an antibody surface can accommodate only one antigen. Here, we describe an antibody with an antigen binding site that binds two distinct proteins with high affinity. We isolated a variant of Herceptin, a therapeutic monoclonal antibody that binds the human epidermal growth factor receptor 2 (HER2), on the basis of its ability to simultaneously interact with vascular endothelial growth factor (VEGF). Crystallographic and mutagenesis studies revealed that distinct amino acids of this antibody, called bH1, engage HER2 and VEGF energetically, but there is extensive overlap between the antibody surface areas contacting the two antigens. An affinity-improved version of bH1 inhibits both HER2- and VEGF-mediated cell proliferation in vitro and tumor progression in mouse models. Such “two-in-one” antibodies challenge the monoclonal antibody paradigm of one binding site, one antigen. They could also provide new opportunities for antibody-based therapy.
A Meta-Selective Copper-Catalyzed C–H Bond Arylation
Authors: Phipps Robert J. , Gaunt Matthew J.
Published: March 2009

For over a century, chemical transformations of benzene derivatives have been guided by the high selectivity for electrophilic attack at the ortho/para positions in electron-rich substrates and at the meta position in electron-deficient molecules. We have developed a copper-catalyzed arylation reaction that, in contrast, selectively substitutes phenyl electrophiles at the aromatic carbon–hydrogen sites meta to an amido substituent. This previously elusive class of transformation is applicable to a broad range of aromatic compounds.
Potential of Rapid Diagnosis for Controlling Drug-Susceptible and Drug-Resistant Tuberculosis in Communities Where
Mycobacterium tuberculosis
Infections Are Highly Prevalent
Authors: Uys Pieter W. , Warren Robin , van Helden Paul D. , Murray Megan , Victor Thomas C.
Published: March 2009

The long-term persistence of
Mycobacterium tuberculosis
in communities with high tuberculosis prevalence is a serious problem aggravated by the presence of drug-resistant tuberculosis strains. Drug resistance in an individual patient is often discovered only after a long delay, particularly if the diagnosis is based on current culture-based drug sensitivity testing methods. During such delays, the patient may transmit tuberculosis to his or her contacts. Rapid diagnosis of drug resistance would be expected to reduce this transmission and hence to decrease the prevalence of drug-resistant strains. To investigate this quantitatively, a mathematical model was constructed, assuming a homogeneous population structure typical of communities in South Africa where tuberculosis incidence is high. Computer simulations performed with this model showed that current control strategies will not halt the spread of multidrug-resistant tuberculosis in such communities. The simulations showed that the rapid diagnosis of drug resistance can be expected to reduce the incidence of drug-resistant cases provided the additional measure of screening within the community is implemented.
Microbiological and Epidemiological Features of Clinical Respiratory Isolates of
Burkholderia gladioli
Authors: Segonds Christine , Clavel-Batut Patricia , Thouverez Michelle , Grenet Dominique , Le Coustumier Alain , Plésiat Patrick , Chabanon Gérard
Published: March 2009

Burkholderia gladioli
, primarily known as a plant pathogen, is involved in human infections, especially in patients with cystic fibrosis (CF). In the present study, the first respiratory isolates recovered from 14 French patients with CF and 4 French patients without CF, identified by 16S rRNA gene analysis, were tested for growth on
B. cepacia
selective media, for identification by commercial systems, and for their antimicrobial susceptibilities, and were compared by pulsed-field gel electrophoresis (PFGE). Patients' data were collected. All 18 isolates grew on oxidation-fermentation-polymyxin B-bacitracin-lactose medium and
Pseudomonas cepacia
agar, but only 13 grew on
Burkholderia cepacia
selective agar. API 20NE strips did not differentiate
B. gladioli
from
B. cepacia
, whereas Vitek 2 GN cards correctly identified 15 isolates. All isolates were susceptible to piperacillin, imipenem, aminoglycosides, and ciprofloxacin and were far less resistant to ticarcillin than
B. cepacia
complex organisms. Fifteen PFGE types were observed among the 18 isolates, but shared types were not identified among epidemiologically related patients. The microbiological follow-up of CF patients showed that colonization was persistent in 3 of 13 documented cases;
B. gladioli
was isolated from posttransplantation cultures of blood from 1 patient. Among the patients without CF,
B. gladioli
was associated with intubation (three cases) or bronchiectasis (one case). In summary, the inclusion of
B. gladioli
in the databases of commercial identification systems should improve the diagnostic capabilities of those systems. In CF patients, this organism is more frequently involved in transient infections than in chronic infections, but it may be responsible for complications posttransplantation; patient-to-patient transmission has not been demonstrated to date. Lastly,
B. gladioli
appears to be naturally susceptible to aminoglycosides and ciprofloxacin, although resistant isolates may emerge in the course of chronic infections.
Identification of Integrin α3 as a New Substrate of the Adenovirus E4orf6/E1B 55-Kilodalton E3 Ubiquitin Ligase Complex
Authors: Dallaire Frédéric , Blanchette Paola , Groitl Peter , Dobner Thomas , Branton Philip E.
Published: March 2009

The human adenovirus E4orf6 and E1B55K proteins promote viral replication by targeting several cellular proteins for degradation. The E4orf6 product has been shown by our group and others to form an E3 ubiquitin ligase complex that contains elongins B and C and cullin family member Cul5. E1B55K associates with this complex, where it is believed to function primarily to introduce bound substrates for degradation via proteasomes. In addition to p53, its first known substrate, the E4orf6/E1B 55-kDa complex (E4orf6/E1B55K) was shown to promote the degradation of Mre11 and DNA ligase IV; however, additional substrates are believed to exist. This notion is strengthened by the fact that none of these substrates seems likely to be associated with additional functions shown to be mediated by the E4orf6-associated E3 ubiquitin ligase complex, including export of late viral mRNAs and blockage of export of the bulk cellular mRNAs from the nucleus. In an attempt to identify new E4orf6/E1B55K substrates, we undertook a proteomic screen using human p53-null, non-small-cell lung carcinoma H1299 cells expressing either E4orf6 protein alone or in combination with E1B55K through infection by appropriate adenovirus vectors. One cellular protein that appeared to be degraded by E1B55K in combination with the E4orf6 protein was a species of molecular mass ∼130 kDa that was identified as the integrin α3 subunit (i.e., very late activation antigen 3 alpha subunit). Preliminary analyses suggested that degradation of α3 may play a role in promoting release and spread of progeny virions.
Differential Effect of Human Herpesvirus 6A on Cell Division and Apoptosis among Naïve and Central and Effector Memory CD4
+
and CD8
+
T-Cell Subsets
Authors: Gupta Sudhir , Agrawal Sudhanshu , Gollapudi Sastry
Published: March 2009

The immune responses of naive and different memory subsets of CD4
+
and CD8
+
T cells to human herpesvirus 6 (HHV-6) have not been previously investigated. We show that HHV-6A induces cell division, as measured by 5,6-carboxyfluorescein succinimidyl ester dye and flow cytometry, predominantly in two populations of effector memory CD4
+
and CD8
+
T cells (T
EM
and T
EMRA
); naïve (T
N
) and central memory (T
CM
) CD4
+
and CD8
+
T cells showed almost no cell division. In contrast, HHV-6A induced apoptosis primarily in T
N
and T
CM
CD4
+
and CD8
+
T cells, whereas T
EM
and T
EMRA
CD4
+
and CD8
+
T cells were resistant to HHV-6A-induced apoptosis. HHV-6A-induced apoptosis was associated with activation of caspase-8, caspase-9, and caspase-3, suggesting the involvement of death receptor and mitochondrial signaling pathways. In addition, HHV-6A induced secretion of interleukin-6 (IL-6), tumor necrosis factor alpha (TNF-α), IL-8, and gamma interferon by peripheral blood mononuclear cells; TNF-α secretion was observed exclusively from CCR7
+
(T
N
plus T
CM
) CD4
+
T cells. These data show that HHV-6 differentially influences the functions of naïve T cells and different subsets of memory CD4
+
and CD8
+
T cells, which in part may be due to differential susceptibility to HHV-6A-induced apoptosis.
Human Immunodeficiency Virus Type 1 gp120 Reprogramming of CD4+T-Cell Migration Provides a Mechanism for Lymphadenopathy
Authors: Green Daniel S. , Center David M. , Cruikshank William W.
Published: March 2009

Infection by human immunodeficiency virus type 1 (HIV-1) is associated with decreases in peripheral CD4+T cells and development of lymphadenopathy. The precise mechanisms by which HIV-1 induces these changes have not been elucidated. T-cell trafficking through lymphoid tissues is facilitated by CCL21-mediated entry and sphingosine-1-phosphate (S1P)-mediated egress. Having previously determined that HIV-1 envelop glycoprotein, gp120, directly alters T-cell migration, we investigated whether gp120 without HIV-1 infection could influence the responses of CD4+T cells to the signals involved in T-cell trafficking through lymph tissue. Incubation of normal human T cells with gp120 for 1 h resulted in reprogramming of CD4 T-cell migratory responses by increasing sensitivity to CCL20 and CCL21 and complete inhibition of migration to S1P. Incubation of human T cells with gp120 prior to injection into NOD.CB17-Prkdcscid/J mice resulted in increases in lymph node accumulation of CD4+T cells, with reciprocal decreases in blood and spleen compared to T cells not exposed to gp120. The effects of gp120 required CD4 signaling mediated through p56lck. These findings suggest that gp120 alone can alter CD4+influx and efflux from lymph nodes in a fashion consistent with the development of lymphopenia and lymphadenopathy.
Broad Tissue and Cell Tropism of Avian Bornavirus in Parrots with Proventricular Dilatation Disease
Authors: Rinder Monika , Ackermann Andreas , Kempf Hermann , Kaspers Bernd , Korbel Rüdiger , Staeheli Peter
Published: March 2009

Avian bornaviruses (ABV), representing a new genus within the family
Bornaviridae
, were recently discovered in parrots from North America and Israel with proventricular dilatation disease (PDD). We show here that closely related viruses are also present in captive European parrots of various species with PDD. The six ABV strains that we identified in clinically diseased birds are new members of the previously defined ABV genotypes 2 and 4. Viruses of both genotypes readily established persistent, noncytolytic infections in quail and chicken cell lines but did not grow in cultured mammalian cells in which classical Borna disease virus strains replicate very efficiently. ABV antigens were present in both the cytoplasm and nucleus of infected cells, suggesting nuclear replication of ABV. The genome organization of avian and mammalian bornaviruses is highly conserved except that ABV lacks a distinct control element in the 5′ noncoding region of the bicistronic mRNA encoding the viral proteins X and P. Reverse transcription-PCR analysis demonstrated the presence of virus in many, if not all, organs of birds with PDD. Viral nucleic acid was also found in feces of diseased birds, suggesting virus transmission by the fecal-oronasal route. Immunohistochemical analysis of organs from birds with PDD revealed that infection with ABV is not restricted to cells of the nervous system. Thus, ABV exhibits a broad tissue and cell tropism that is strikingly different from classical Borna disease virus.
CCR5- and CXCR4-Tropic Subtype C Human Immunodeficiency Virus Type 1 Isolates Have a Lower Level of Pathogenic Fitness than Other Dominant Group M Subtypes: Implications for the Epidemic
Authors: Abraha Awet , Nankya Immaculate L. , Gibson Richard , Demers Korey , Tebit Denis M. , Johnston Elizabeth , Katzenstein David , Siddiqui Asna , Herrera Carolina , Fischetti Lucia , Shattock Robin J. , Arts Eric J.
Published: March 2009

Human immunodeficiency virus type 1 (HIV-1) subtype C is the dominant subtype globally, due largely to the incidence of subtype C infections in sub-Saharan Africa and east Asia. We compared the relative replicative fitness (ex vivo) of the major (M) group of HIV-1 subtypes A, B, C, D, and CRF01_AE and group O isolates. To estimate pathogenic fitness, pairwise competitions were performed between CCR5-tropic (R5) or CXCR4-tropic (X4) virus isolates in peripheral blood mononuclear cells (PBMC). A general fitness order was observed among 33 HIV-1 isolates; subtype B and D HIV-1 isolates were slightly more fit than the subtype A and dramatically more fit than the 12 subtype C isolates. All group M isolates were more fit (ex vivo) than the group O isolates. To estimate ex vivo transmission fitness, a subset of primary HIV-1 isolates were examined in primary human explants from penile, cervical, and rectal tissues. Only R5 isolates and no X4 HIV-1 isolates could replicate in these tissues, whereas the spread to PM1 cells was dependent on active replication and passive virus transfer. In tissue competition experiments, subtype C isolates could compete with and, in some cases, even win over subtype A and D isolates. However, when the migratory cells from infected tissues were mixed with a susceptible cell line, the subtype C isolates were outcompeted by other subtypes, as observed in experiments with PBMC. These findings suggest that subtype C HIV-1 isolates might have equal transmission fitness but reduced pathogenic fitness relative to other group M HIV-1 isolates.
A Novel Cardiotropic Murine Adenovirus Representing a Distinct Species of Mastadenoviruses
Authors: Klempa Boris , Krüger Detlev H. , Auste Brita , Stanko Michal , Krawczyk Adalbert , Nickel Katrin F. , Überla Klaus , Stang Alexander
Published: March 2009

During cell culture isolation experiments to recover Dobrava hantavirus from a suspension of liver from a striped field mouse (
Apodemus agrarius
), an unknown virus was coisolated. Atypically for hantaviruses, it had extensive cytopathic effects. Using a random PCR approach, it was identified as a novel murine adenovirus, MAdV-3 (for MAdV type 3). A plaque-purified virus clone was prepared and further characterized. The complete genome sequence of MAdV-3 was determined to be 30,570 bp in length. Sequence comparisons to other adenovirus species revealed highest similarity to MAdV-1, the representative of the murine adenovirus A species. However, substantial differences were found in the E1, E3, and E4 genomic regions. The phylogenetic distance of MAdV-3 amino acid sequences for pVIII, protease, polymerase, and hexon from MAdV-1 is markedly higher than 0.1 exchange per position, and, based on our cross-neutralization experiments, MAdV-3 and MAdV-1 can be regarded as different serotypes. Therefore, we propose to classify MAdV-3 as the first isolate of a novel adenovirus species, designated murine adenovirus C (MAdV-C). The novel MAdV-3 virus is not only genetically and serologically distinct from MAdV-1 but also shows a unique organ tropism in infected mice. In contrast to MAdV-1, the virus was not detectable in brain but predominantly infected heart tissue. Thus, infection of mice with cardiotropic MAdV-3 might be an interesting animal model of adenovirus-induced myocarditis.
Role of N-Linked Glycosylation for Sindbis Virus Infection and Replication in Vertebrate and Invertebrate Systems
Authors: Knight Ronald L. , Schultz Kimberly L. W. , Kent Rebekah J. , Venkatesan Meera , Griffin Diane E.
Published: March 2009

Each Sindbis virus (SINV) surface glycoprotein has two sites for N-linked glycosylation (E1 positions 139 and 245 [E1-139 and E1-245] and E2 positions 196 and 318 [E2-196 and E2-318]). Studies of SINV strain TE12 mutants with each site eliminated identified the locations of carbohydrates by cryo-electron microscopy (S. V. Pletnev et al., Cell 105:127-136, 2001). In the current study, the effects of altered glycosylation on virion infectivity, growth in cells of vertebrates and invertebrates, heparin binding, virulence in mice, and replication in mosquitoes were assessed. Particle-to-PFU ratios for E1-139 and E2-196 mutant strains were similar to that for TE12, but this ratio for the E1-245 mutant was 100-fold lower than that for TE12. Elimination of either E2 glycosylation site increased virus binding to heparin and increased replication in BHK cells. Elimination of either E1 glycosylation site had no effect on heparin binding but resulted in an approximately 10-fold decrease in virus yield from BHK cells compared to the TE12 amount. No differences in pE2 processing were detected. E2-196 and E2-318 mutants were more virulent in mice after intracerebral inoculation, while E1-139 and E1-245 mutants were less virulent. The E1-245 mutant showed impaired replication in C7/10 mosquito cells and in
Culex quinquefasciatus
after intrathoracic inoculation. We conclude that the increased replication and virulence of E2-196 and E2-318 mutants are primarily due to increased efficiency of binding to heparan sulfate on mammalian cells. Lack of glycosylation at E1-139 or E1-245 impairs replication in vertebrate cells, while E1-245 also severely affects replication in invertebrate cells.
Cell-to-Cell Spread of the RNA Interference Response Suppresses Semliki Forest Virus (SFV) Infection of Mosquito Cell Cultures and Cannot Be Antagonized by SFV
Authors: Attarzadeh-Yazdi Ghassem , Fragkoudis Rennos , Chi Yi , Siu Ricky W. C. , Ülper Liane , Barry Gerald , Rodriguez-Andres Julio , Nash Anthony A. , Bouloy Michèle , Merits Andres , Fazakerley John K. , Kohl Alain
Published: March 2009

In their vertebrate hosts, arboviruses such as Semliki Forest virus (SFV) (
Togaviridae
) generally counteract innate defenses and trigger cell death. In contrast, in mosquito cells, following an early phase of efficient virus production, a persistent infection with low levels of virus production is established. Whether arboviruses counteract RNA interference (RNAi), which provides an important antiviral defense system in mosquitoes, is an important question. Here we show that in
Aedes albopictus
-derived mosquito cells, SFV cannot prevent the establishment of an antiviral RNAi response or prevent the spread of protective antiviral double-stranded RNA/small interfering RNA (siRNA) from cell to cell, which can inhibit the replication of incoming virus. The expression of tombusvirus siRNA-binding protein p19 by SFV strongly enhanced virus spread between cultured cells rather than virus replication in initially infected cells. Our results indicate that the spread of the RNAi signal contributes to limiting virus dissemination.
Prion Protein Glycosylation Is Not Required for Strain-Specific Neurotropism
Authors: Piro Justin R. , Harris Brent T. , Nishina Koren , Soto Claudio , Morales Rodrigo , Rees Judy R. , Supattapone Surachai
Published: March 2009

In this study, we tested the hypothesis that the glycosylation of the pathogenic isoform of the prion protein (PrP
Sc
) might encode the selective neurotropism of prion strains. We prepared unglycosylated cellular prion protein (PrP
C
) substrate molecules from normal mouse brain by treatment with PNGase F and used reconstituted serial protein cyclic misfolding amplification reactions to produce RML and 301C mouse prions containing unglycosylated PrP
Sc
molecules. Both RML- and 301C-derived prions containing unglycosylated PrP
Sc
molecules were infectious to wild-type mice, and neuropathological analysis showed that mice inoculated with these samples maintained strain-specific patterns of PrP
Sc
deposition and neuronal vacuolation. These results show that PrP
Sc
glycosylation is not necessary for strain-dependent prion neurotropism.
Identification of an Intercistronic Internal Ribosome Entry Site in a Marek's Disease Virus Immediate-Early Gene
Authors: Tahiri-Alaoui Abdessamad , Smith Lorraine P. , Baigent Suzan , Kgosana Lydia , Petherbridge Lawrence J. , Lambeth Luke S. , James William , Nair Venugopal
Published: March 2009

In this study, we have identified an internal ribosome entry site (IRES) from the highly infectious herpesvirus Marek's disease virus (MDV). The IRES was mapped to the intercistronic region (ICR) of a bicistronic mRNA that we cloned from the MDV-transformed CD4
+
T-cell line MSB-1. The transcript is a member of a family of mRNAs expressed as immediate-early genes with two open reading frames (ORF). The first ORF encodes a 14-kDa polypeptide with two N-terminal splice variants, whereas the second ORF is contained entirely within a single exon and encodes a 12-kDa protein also known as RLORF9. We have shown that the ICR that separates the two ORFs functions as an IRES that controls the translation of RLORF9 when cap-dependent translation is inhibited. Deletion analysis revealed that there are two potential IRES elements within the ICR. Reverse genetic experiments with the oncogenic strain of MDV type 1 indicated that deletion of IRES-controlled RLORF9 does not significantly affect viral replication or MDV-induced mortality.
Design procedure for vertical drains considering a linear variation of lateral permeability within the smear zone
Authors: Rujikiatkamjorn Cholachat , Indraratna Buddhima
Published: March 2009

A system of vertical drains with surcharge preloading is an effective method for promoting radial drainage and accelerated soil consolidation. This study presents a procedure for the design of vertical drains that significantly extends the previous technique proposed by the authors to include: (i) a linear reduction of lateral permeability in the smear zone, (ii) the effect of overlapping smear zones in a closely spaced drain network, and (iii) the gain in undrained shear strength due to consolidation. Design examples are provided for both single stage and multi-stage embankment construction demonstrating the convenient use of the proposed solutions in practical situations.
Progressive polyuria without vasopressin neuron loss in a mouse model for familial neurohypophysial diabetes insipidus
Authors: Hayashi Masayuki , Arima Hiroshi , Ozaki Noriyuki , Morishita Yoshiaki , Hiroi Maiko , Ozaki Nobuaki , Nagasaki Hiroshi , Kinoshita Noriaki , Ueda Masatsugu , Shiota Akira , Oiso Yutaka
Published: March 2009

Familial neurohypophysial diabetes insipidus (FNDI), an autosomal dominant disorder, is mostly caused by mutations in the gene of neurophysin II (NPII), the carrier protein of arginine vasopressin (AVP). Previous studies suggest that loss of AVP neurons might be the cause of polyuria in FNDI. Here we analyzed knockin mice expressing mutant NPII that causes FNDI in humans. The heterozygous mice manifested progressive polyuria as do patients with FNDI. Immunohistochemical analyses revealed that inclusion bodies that were not immunostained with antibodies for mutant NPII, normal NPII, or AVP were present in the AVP cells in the supraoptic nucleus (SON), and that the size of inclusion bodies gradually increased in parallel with the increases in urine volume. Electron microscopic analyses showed that aggregates existed in the endoplasmic reticulum (ER) as well as in the nucleus of AVP neurons in 1-mo-old heterozygous mice. At 12 mo, dilated ER filled with aggregates occupied the cytoplasm of AVP cells, while few aggregates were found in the nucleus. Analyses with in situ hybridization revealed that expression of AVP mRNA was significantly decreased in the SON in the heterozygous mice compared with that in wild-type mice. Counting cells expressing AVP mRNA in the SON indicated that polyuria had progressed substantially in the absence of neuronal loss. These data suggest that cell death is not the primary cause of polyuria in FNDI, and that the aggregates accumulated in the ER might be involved in the dysfunction of AVP neurons that lead to the progressive polyuria.