bluuebunny/crossref_metadata_2025 huggingface dataset:
Persistent altered fusion transcript splicing identifies RUNX1‐RUNX1T1+ AML patients likely to relapse
Authors: Ommen Hans B. , Østergaard Mette , Yan Ming , Brændstrup Karin , Zhang Dong‐Er , Hokland Peter
Published: November 2009

In acute myeloid leukemia (AML) mouse models, the RUNX1‐RUNX1T1 fusion protein has failed to produce leukemia by itself, but alternative splicing of exon 9a of the RUNX1‐RUNX1T1 fusion transcript (FT) has recently been shown to enhance the leukemogenic potential. We have analyzed 138 diagnosis and follow‐up samples from 13 RUNX1‐RUNX1T1+ patients as well as diagnosis samples from 13 RUNX1‐RUNX1T1− AML patients and 26 healthy donors. Levels of native RUNX1T1 mRNA were low in both healthy and RUNX1‐RUNX1T1‐negative AML samples. Likewise, the ratio between RUNX1T1 mRNA harboring exon 9a and lacking exon 9a was low and tightly regulated (0.017–0.11). In contrast, 11/13 RUNX1‐RUNX1T1‐positive AML patients displayed high and variable ratios of FT ranging from 0.05 to 0.46 (P < 0.001, Wilcoxon rank‐sum test), indicating altered exon 9a splicing in these patients. Importantly, patients who remained in continuous complete remission displayed a faster disappearance of the RUNX1‐RUNX1T1 exon 9a splice variant compared to patients bound to relapse (P = 0.02). In conclusion, alternative splicing seems to be part of the leukemogenic process in the majority of RUNX1‐RUNX1T1‐positive AML patients, and splice variant kinetics under cytoreduction may be a predictor for patients prone to relapse.
Intertask comparison of frontal plane knee position and moment in female athletes during three distinct movement tasks
Authors: Harty C. M. , DuPont C. E. , Chmielewski T. L. , Mizner R. L.
Published: November 2009

Gender differences in dynamic frontal plane knee posture during functional tasks contribute to increased anterior cruciate ligament injury risk in female athletes. Many tasks have been used to assess frontal plane movement patterns, but few studies compare patterns across tasks or evaluate the influence of static alignment on dynamic postures. The purpose of our study was to (1) establish the intertask differences in frontal plane knee posture during step down, single leg land, and drop vertical jump tasks; (2) determine intra‐athlete correlations in knee posture across tasks; and (3) investigate the intra‐athlete correlations between frontal plane knee posture while standing and during movement. Thirty‐seven female athletes from high demand sports participated in a motion analysis assessment. Substantial differences in the frontal plane knee angles and moments occurred between tasks. However, there were strong relationships in frontal plane knee angles (r=0.72–0.76) and moderate to good relationships in knee moments across tasks (r=0.426–0.627). No relationship existed between the standing and dynamic knee abduction angles (r<0.06). Female athletes who demonstrate at‐risk knee position and loading are likely to do so across tasks. The lack of relationship between static and dynamic frontal plane knee posture suggests that neuromuscular mechanisms contribute to the consistency in movement patterns across tasks.
Online activities of urban Malaysian adolescents: report of a pilot study
Authors: Tan Kok Eng , Yen Abdullah Melissa Ng Lee , Guan Saw Kim
Published: November 2009

The literacy practices of many communities today show new ways of meaning making in the contemporary, technological and digital culture. A number of Malaysian adolescents belong to this culture. This pilot study reports the preliminary findings of a larger study aimed at describing the online activities of Malaysian adolescents. Fifty‐four participants from an urban secondary school in Penang took part in this study. One finding showed that the percentages of language use on the Internet were: 80.6 per cent in English, 10.6 per cent in Bahasa Malaysia and 8.8 per cent in Chinese. In addition, the study also revealed some common websites that these adolescents frequented. The findings point to the need to investigate the extent of these online literacy practices beyond school among the adolescent population in Malaysia and to look into the implications for ESL education.
Depression Among a Sample of First‐Time Adolescent and Adult Mothers
Authors: Lanzi Robin Gaines , Bert Shannon Carothers , Jacobs Bette Keltner ,   the Centers for the Prevention of Child Neglect
Published: November 2009

PROBLEM:  Little is known about the rates, correlates, and consequences of depression among a sample of first‐time mothers.

METHODS:  Four‐site prospective study of the first 3 years of life among first children born to first‐time mothers, including adolescent (N= 396), lower‐ (N= 169), and higher‐resource adult (N= 117). Mothers were administered the Beck Depression Inventory prenatally and 6 months postpartum. Measures of maternal and infant behaviors were taken at 8 months.

FINDINGS:  Adolescent mothers displayed higher prenatal and 6‐month rates of depression than lower‐ and higher‐resource adult mothers, with significantly more adolescent mothers “consistently” depressed at the two time points than lower‐ and higher‐resource adult mothers. Further, adolescent mothers were significantly more likely to become depressed after their babies were born than lower‐ or higher‐resource adult mothers. Depression was negatively related to parenting practices and babies' behavior. As depression increased, mothers scored less favorably in maternal warmth and sensitivity, contingent responsiveness, and general verbalness; children scored less favorably in warmth seeking toward their mothers.

CONCLUSION:  Findings signify the need for counseling and nurse‐based intervention and prevention services geared at preparing pregnant adolescents for motherhood.
Modeling Surface Water‐Groundwater Interaction with MODFLOW: Some Considerations
Authors: Brunner Philip , Simmons Craig T. , Cook Peter G. , Therrien René
Published: November 2009

The accuracy with which MODFLOW simulates surface water‐groundwater interaction is examined for connected and disconnected losing streams. We compare the effect of different vertical and horizontal discretization within MODFLOW and also compare MODFLOW simulations with those produced by HydroGeoSphere. HydroGeoSphere is able to simulate both saturated and unsaturated flow, as well as surface water, groundwater and the full coupling between them in a physical way, and so is used as a reference code to quantify the influence of some of the simplifying assumptions of MODFLOW. In particular, we show that (1) the inability to simulate negative pressures beneath disconnected streams in MODFLOW results in an underestimation of the infiltration flux; (2) a river in MODFLOW is either fully connected or fully disconnected, while in reality transitional stages between the two flow regimes exist; (3) limitations in the horizontal discretization of the river can cause a mismatch between river width and cell width, resulting in an error in the water table position under the river; and (4) because coarse vertical discretization of the aquifer is often used to avoid the drying out of cells, this may result in an error in simulating the height of the groundwater mound. Conditions under which these errors are significant are investigated.
Corneal hysteresis measured with the Ocular Response Analyzer® in normal and glaucomatous eyes
Authors: Abitbol Olivia , Bouden Jihène , Doan Serge , Hoang‐Xuan Thanh , Gatinel Damien
Published: November 2009

Purpose:  To identify differences in corneal hysteresis (CH) and central corneal thickness (CCT) between healthy and glaucomatous patients.

Methods:  Retrospective observational study. One hundred and thirty‐three eyes of 75 healthy and 58 glaucomatous patients were included. CH was measured in each patient using Ocular Response Analyzer. CCT was determined by ultrasonic pachymetry. For each patient, one eye was randomly selected. We used a Student t‐test to search for significant differences between the different groups (p<0.05).

Results:  In healthy and glaucomatous eyes, mean CH values were 10.46 ± 1.6 and 8.77 ± 1.4 mm Hg, respectively. Mean CCT values were 560.2 ± 36.3 and 535.3 ± 42.7 μm, respectively. CH and CCT were significantly lower in glaucomatous eyes than in normal eyes, (p<0.05).

Discussion:  In our series, CH was lower in glaucomatous than in normal eyes. The relationship between glaucoma, IOP, and ocular structures may not be confined to the consideration of CCT. A low CH value could be responsible for under‐estimation of IOP. CH could also be a risk factor for glaucoma, independent of IOP. Further studies are needed to support these hypotheses.

Conclusion:  In our investigation, CCT and CH were significantly lower in glaucomatous eyes than in healthy eyes.
A Translational Research Niche for Small Business Innovation Research Grants
Authors: Handelsman Karl
Published: November 2009

SBIR funding can be part of the health care solution by financing small
biomedical companies in a way that stokes their translational engines.
Translational Medicine: An Engine of Change for Bringing New Technology to Community Health
Authors: Milne Christopher-Paul , Kaitin Kenneth I.
Published: November 2009

A patient-centered focus in R&D should lead to improvements in medical care.
Cancer Transformers
Authors: Khidr Lily
Published: November 2009

Environmental factors such as inflammation can trip an epigenetic switch that causes self-propelling cancer progression, without mutations in tumor suppressor genes or oncogenes.
Physical and Chemical Characterization and Immunologic Properties ofSalmonella entericaSerovar Typhi Capsular Polysaccharide-Diphtheria Toxoid Conjugates
Authors: Cui Changfa , Carbis Rodney , An So Jung , Jang Hyun , Czerkinsky Cecil , Szu Shousun C. , Clemens John D.
Published: November 2009

Typhoid fever remains a serious public health problem in developing countries, especially among young children. Recent studies showed more than 50% of typhoid cases are in children under 5 years old. Licensed vaccines, such asSalmonella entericaserovar Typhi capsular Vi, did not confer protection against typhoid fever for this age group. Vi conjugate, prepared by binding Vi toPseudomonas aeruginosarecombinant exoprotein A (rEPA), induces protective levels of antibody at as young as 2 years old. Because of the lack of regulatory precedent for rEPA in licensing vaccines, we employed diphtheria toxoid (DT) as the carrier protein to accommodate accessibility in developing countries. Five lots of Vi-DT conjugates were prepared using adipic acid dihydrazide (ADH) as the linker. All 5 lots showed consistency in their physical and chemical characteristics and final yields. These Vi-DT conjugates elicited levels of IgG anti-Vi in young mice significantly higher than those in mice injected with Vi alone and induced a booster response upon reinjection. This booster effect was absent if the Vi replaced one of the two conjugate injections. Vi-DT was stable under repeated freeze-thaw (20 cycles). We plan to perform clinical evaluation of the safety and immunogenicity of Vi-DT when added to the infant combination vaccines.
Diversity of
Staphylococcus
Species Strains Based on Partial
kat
(Catalase) Gene Sequences and Design of a PCR-Restriction Fragment Length Polymorphism Assay for Identification and Differentiation of Coagulase-Positive Species (
S. aureus
,
S. delphini
,
S. hyicus
,
S. intermedius
,
S. pseudintermedius
, and
S. schleiferi
subsp.
coagulans
)
Authors: Blaiotta Giuseppe , Fusco Vincenzina , Ercolini Danilo , Pepe Olimpia , Coppola Salvatore
Published: November 2009

A set of degenerate PCR primers was designed and used to amplify and sequence about 75% of the catalase (
kat
) gene from each of 49 staphylococcal strains. In some strains of
Staphylococcus xylosus
,
S. saprophyticus
, and
S. equorum
, two catalase genes,
kat
A
and
kat
B
, were found. A phylogenetic tree was generated and showed diversities among 66 partial (about 900-bp) staphylococcal
kat
nucleotide sequences (including 17 sequences found in GenBank) representing 26 different species. The topology of this tree showed a distribution of staphylococcal species similar, but not identical, to those reported previously based on 16S rRNA,
hsp
60
,
sod
A
,
rpo
B
,
tuf
, and
gap
genes. The
kat
gene sequences were less conserved than those of 16S rRNA,
rpo
B
,
hsp
60
, and
tuf
genes and slightly more conserved than those of the
gap
gene. Therefore,
kat
gene sequence analysis may provide an additional marker for inferring phylogenetic relationships of staphylococci. Moreover, the discrete nucleotide polymorphism revealed in this gene could be exploited for rapid, low-cost identification of staphylococcal species through PCR-restriction fragment length polymorphism (RFLP) analysis. In this study, a PCR-RFLP assay performed by using only the TaqI restriction enzyme was successfully developed for rapid unequivocal identification/differentiation, at species and subspecies levels, of coagulase-positive staphylococci (CPS). The assay was validated by testing the DNA from 100 staphylococcal strains, including reference and wild CPS strains isolated from different environments. This reliable, rapid, and low-cost approach (requiring about 6 h from DNA isolation to the achievement of results and <5 Euros for each strain tested) allowed unambiguous identification of all the strains assayed, including the newly described
S. delphini
and
S. pseudintermedius
CPS species.
An Unusual Pneumococcal Sequence Type Is the Predominant Cause of Serotype 3 Invasive Disease in South Africa
Authors: Mothibeli Kedibone M. , du Plessis Mignon , von Gottberg Anne , de Gouveia Linda , Adrian Peter , Madhi Shabir A. , Klugman Keith P.
Published: November 2009

We reviewed pneumococcal serotype 3 cases reported from 2000 through 2005 to a laboratory-based surveillance system for invasive pneumococcal disease in South Africa. The prevalence of serotype 3 invasive isolates was compared to their prevalence in carriage isolates to determine the odds of invasiveness due to serotype 3 among South African children. Three groups of serotype 3 strains were characterized by pulsed-field gel electrophoresis (PFGE) or Box element PCR (BOX-PCR), randomly selected invasive isolates from one province, isolates from a carriage study involving children in the same province, and antimicrobial-resistant invasive isolates collected nationally. Examples of the PFGE types identified were further characterized by multilocus sequence typing. In total, 15,980 viable isolates causing invasive disease were submitted, of which 661 (4%) were serotype 3, mostly from adults (85% [489/575]). Fewer serotype 3 isolates were nonsusceptible to antimicrobial agents tested (40/661 [6%]) than non-serotype 3 isolates (8,480/15,319 [55%]) (
P
< 0.001). Compared to non-serotype 3 cases, there was no association with HIV coinfection (2,212/2,569 [86%] versus 72/78 [92%];
P
= 0.1) or increased case fatality ratio (1,190/4,211 [28%] versus 54/154 [35%];
P
= 0.7). Serotype 3 in children had a low but statistically insignificant invasive disease potential (odds ratio [OR] of 0.15; 95% confidence interval [CI] of 0.01 to 1.06). Strains were grouped into 3 PFGE clusters, with the largest, cluster A, representing 54% (84/155), including 14 isolates confirmed as sequence type 458 (ST458). It was confirmed that 3 isolates from cluster B, which represented only 12% (18/155) of the isolates, were the serotype 3 global strain, ST180. We have therefore identified ST458 as predominating in South Africa, but with an invasive potential similar to that of the predominant global clone ST180.
Campylobacter fetus
Peritonitis and Bacteremia in a Patient Undergoing Continuous Ambulatory Peritoneal Dialysis
Authors: Romero Gómez M. P. , García-Perea A. , Ruiz Carrascoso G. , Bajo M. A. , Mingorance J.
Published: November 2009

We report a case of
Campylobacter fetus
peritonitis and bacteremia in a patient undergoing continuous ambulatory peritoneal dialysis.
Nonstructural NSs Protein of Rift Valley Fever Virus Interacts with Pericentromeric DNA Sequences of the Host Cell, Inducing Chromosome Cohesion and Segregation Defects
Authors: Mansuroglu Z. , Josse T. , Gilleron J. , Billecocq A. , Leger P. , Bouloy M. , Bonnefoy E.
Published: November 2009

Rift Valley fever virus (RVFV) is an emerging, highly pathogenic virus; RVFV infection can lead to encephalitis, retinitis, or fatal hepatitis associated with hemorrhagic fever in humans, as well as death, abortions, and fetal deformities in animals. RVFV nonstructural NSs protein, a major factor of the virulence, forms filamentous structures in the nuclei of infected cells. In order to further understand RVFV pathology, we investigated, by chromatin immunoprecipitation, immunofluorescence, fluorescence in situ hybridization, and confocal microscopy, the capacity of NSs to interact with the host genome. Our results demonstrate that even though cellular DNA is predominantly excluded from NSs filaments, NSs interacts with some specific DNA regions of the host genome such as clusters of pericentromeric γ-satellite sequence. Targeting of these sequences by NSs was correlated with the induction of chromosome cohesion and segregation defects in RVFV-infected murine, as well as sheep cells. Using recombinant nonpathogenic virus rZHΔNSs210-230, expressing a NSs protein deleted of its region of interaction with cellular factor SAP30, we showed that the NSs-SAP30 interaction was essential for NSs to target pericentromeric sequences, as well as for induction of chromosome segregation defects. The effect of RVFV upon the inheritance of genetic information is discussed with respect to the pathology associated with fetal deformities and abortions, highlighting the main role played by cellular cofactor SAP30 on the establishment of NSs interactions with host DNA sequences and RVFV pathogenesis.
A Global Analysis of Evolutionary Conservation among Known and Predicted Gammaherpesvirus MicroRNAs
Authors: Walz Nicole , Christalla Thomas , Tessmer Uwe , Grundhoff Adam
Published: November 2009

MicroRNAs (miRNAs) are small, noncoding RNAs which posttranscriptionally regulate gene expression. The current release of the miRNA registry lists 16 viruses which encode a total of 146 miRNA hairpins. Strikingly, 139 of these are encoded by members of the herpesvirus family, suggesting an important role for miRNAs in the herpesvirus life cycle. However, with the exception of 7 miRNA hairpins known to be shared by Epstein-Barr virus (EBV) and the closely related rhesus lymphocryptovirus (rLCV), the known herpesvirus miRNAs show little evidence of evolutionary conservation. We have performed a global analysis of miRNA conservation among gammaherpesviruses which is not limited to family members known to encode miRNAs but includes also those which have not been previously analyzed. For this purpose, we have performed a computational prediction of miRNA candidates of all fully sequenced gammaherpesvirus genomes, followed by sequence/structure alignments. Our results indicate that gammaherpesvirus miRNA conservation is limited to two pairs of viral genomes. One is the already-known case of EBV and rLCV. These viruses, however, share significantly more miRNAs than previously thought, as we identified and experimentally verified 10 novel conserved as well as 7 novel nonconserved rLCV pre-miRNA hairpins. The second case consists of rhesus rhadinovirus (RRV), which is predicted to share at least 9 pre-miRNAs with the closely related Japanese macaque herpesvirus (JMHV). Although several other gammaherpesviruses are predicted to encode large numbers of clustered miRNAs at conserved genomic loci, no further examples of evolutionarily conserved miRNA sequences were found.
A Single Amino Acid Substitution in HIV-1 Reverse Transcriptase Significantly Reduces Virion Release
Authors: Chiang Chien-Cheng , Wang Shiu-Mei , Pan Yen-Yu , Huang Kuo-Jung , Wang Chin-Tien
Published: November 2009

HIV-1 protease (PR) mediates the proteolytic processing of virus particles during or after virus budding. PR activation is thought to be triggered by appropriate Gag-Pol/Gag-Pol interaction; factors affecting this interaction either enhance or reduce PR-mediated cleavage efficiency, resulting in markedly reduced virion production or the release of inadequately processed virions. We previously showed that a Gag-Pol deletion mutation involving the reverse transcriptase tryptophan (Trp) repeat motif markedly impairs PR-mediated virus maturation and that an alanine substitution at W401 (W401A) or at both W401 and W402 (W401A/W402A) partially or almost completely negates the enhancement effect of efavirenz (a nonnucleoside reverse transcriptase inhibitor) on PR-mediated virus processing efficiency. These data suggest that the Trp repeat motif may contribute to the PR activation process. Here we demonstrate that due to enhanced Gag cleavage efficiency, W402 alanine or leucine substitution significantly reduces virus production. However, W402 replacement with phenylalanine does not significantly affect virus particle assembly or processing, but it does markedly impair viral infectivity in a single-cycle infection assay. Our results demonstrate that a single amino acid substitution at HIV-1 RT can radically affect virus assembly by enhancing Gag cleavage efficiency, suggesting that in addition to contributing to RT biological function during the early stages of virus replication, the HIV-1 RT tryptophan repeat motif in a Gag-Pol context may play an important role in suppressing the premature activation of PR during late-stage virus replication.
Evolutionary Patterns of Eastern Equine Encephalitis Virus in North versus South America Suggest Ecological Differences and Taxonomic Revision
Authors: Arrigo Nicole C. , Adams A. Paige , Weaver Scott C.
Published: November 2009

The eastern equine encephalitis (EEE) complex consists of four distinct genetic lineages: one that circulates in North America (NA EEEV) and the Caribbean and three that circulate in Central and South America (SA EEEV). Differences in their geographic, pathogenic, and epidemiologic profiles prompted evaluation of their genetic diversity and evolutionary histories. The structural polyprotein open reading frames of all available SA EEEV and recent NA EEEV isolates were sequenced and used in evolutionary and phylogenetic analyses. The nucleotide substitution rate per year for SA EEEV (1.2 × 10−4) was lower and more consistent than that for NA EEEV (2.7 × 10−4), which exhibited considerable rate variation among constituent clades. Estimates of time since divergence varied widely depending upon the sequences used, with NA and SA EEEV diverging ca. 922 to 4,856 years ago and the two main SA EEEV lineages diverging ca. 577 to 2,927 years ago. The single, monophyletic NA EEEV lineage exhibited mainly temporally associated relationships and was highly conserved throughout its geographic range. In contrast, SA EEEV comprised three divergent lineages, two consisting of highly conserved geographic groupings that completely lacked temporal associations. A phylogenetic comparison of SA EEEV and Venezuelan equine encephalitis viruses (VEEV) demonstrated similar genetic and evolutionary patterns, consistent with the well-documented use of mammalian reservoir hosts by VEEV. Our results emphasize the evolutionary and genetic divergences between members of the NA and SA EEEV lineages, consistent with major differences in pathogenicity and ecology, and propose that NA and SA EEEV be reclassified as distinct species in the EEE complex.
The Open Reading Frame 3a Protein of Severe Acute Respiratory Syndrome-Associated Coronavirus Promotes Membrane Rearrangement and Cell Death
Authors: Freundt Eric C. , Yu Li , Goldsmith Cynthia S. , Welsh Sarah , Cheng Aaron , Yount Boyd , Liu Wei , Frieman Matthew B. , Buchholz Ursula J. , Screaton Gavin R. , Lippincott-Schwartz Jennifer , Zaki Sherif R. , Xu Xiao-Ning , Baric Ralph S. , Subbarao Kanta , Lenardo Michael J.
Published: November 2009

The genome of the severe acute respiratory syndrome-associated coronavirus (SARS-CoV) contains eight open reading frames (ORFs) that encode novel proteins. These accessory proteins are dispensable for in vitro and in vivo replication and thus may be important for other aspects of virus-host interactions. We investigated the functions of the largest of the accessory proteins, the ORF 3a protein, using a 3a-deficient strain of SARS-CoV. Cell death of Vero cells after infection with SARS-CoV was reduced upon deletion of ORF 3a. Electron microscopy of infected cells revealed a role for ORF 3a in SARS-CoV induced vesicle formation, a prominent feature of cells from SARS patients. In addition, we report that ORF 3a is both necessary and sufficient for SARS-CoV-induced Golgi fragmentation and that the 3a protein accumulates and localizes to vesicles containing markers for late endosomes. Finally, overexpression of ADP-ribosylation factor 1 (Arf1), a small GTPase essential for the maintenance of the Golgi apparatus, restored Golgi morphology during infection. These results establish an important role for ORF 3a in SARS-CoV-induced cell death, Golgi fragmentation, and the accumulation of intracellular vesicles.
Newcastle Disease Virus-Like Particles Containing Respiratory Syncytial Virus G Protein Induced Protection in BALB/c Mice, with No Evidence of Immunopathology
Authors: Murawski Matthew R. , McGinnes Lori W. , Finberg Robert W. , Kurt-Jones Evelyn A. , Massare Michael J. , Smith Gale , Heaton Penny M. , Fraire Armando E. , Morrison Trudy G.
Published: November 2009

Respiratory syncytial virus (RSV) is the leading cause of serious respiratory infections in children as well as a serious cause of disease in elderly and immunosuppressed populations. There are no licensed vaccines available to prevent RSV disease. We have developed a virus-like particle (VLP) vaccine candidate for protection from RSV. The VLP is composed of the NP and M proteins of Newcastle disease virus (NDV) and a chimeric protein containing the cytoplasmic and transmembrane domains of the NDV HN protein and the ectodomain of the human RSV G protein (H/G). Immunization of mice with 10 or 40 μg total VLP-H/G protein by intraperitoneal or intramuscular inoculation stimulated antibody responses to G protein which were as good as or better than those stimulated by comparable amounts of UV-inactivated RSV. Immunization of mice with two doses or even a single dose of these particles resulted in the complete protection of mice from RSV replication in the lungs. Immunization with these particles induced neutralizing antibodies with modest titers. Upon RSV challenge of VLP-H/G-immunized mice, no enhanced pathology in the lungs was observed, although lungs of mice immunized in parallel with formalin-inactivated RSV (FI-RSV) showed the significant pathology that has previously been documented after immunization with FI-RSV. Thus, the VLP-H/G candidate vaccine was immunogenic in BALB/c mice and prevented replication of RSV in murine lungs, with no evidence of immunopathology. These data support further development of virus-like particle vaccine candidates for protection against RSV.
Analysis of the Viral Elements Required in the Nuclear Import of HIV-1 DNA
Authors: Rivière Lise , Darlix Jean-Luc , Cimarelli Andrea
Published: November 2009

HIV-1 possesses an exquisite ability to infect cells independently from their cycling status by undergoing an active phase of nuclear import through the nuclear pore. This property has been ascribed to the presence of karyophilic elements present in viral nucleoprotein complexes, such as the matrix protein (MA); Vpr; the integrase (IN); and a
cis
-acting structure present in the newly synthesized DNA, the DNA flap. However, their role in nuclear import remains controversial at best. In the present study, we carried out a comprehensive analysis of the role of these elements in nuclear import in a comparison between several primary cell types, including stimulated lymphocytes, macrophages, and dendritic cells. We show that despite the fact that none of these elements is absolutely required for nuclear import, disruption of the central polypurine tract-central termination sequence (cPPT-CTS) clearly affects the kinetics of viral DNA entry into the nucleus. This effect is independent of the cell cycle status of the target cells and is observed in cycling as well as in nondividing primary cells, suggesting that nuclear import of viral DNA may occur similarly under both conditions. Nonetheless, this study indicates that other components are utilized along with the cPPT-CTS for an efficient entry of viral DNA into the nucleus.
Epstein-Barr Viruses That Express a CD21 Antibody Provide Evidence that gp350's Functions Extend beyond B-Cell Surface Binding
Authors: Busse Clemens , Feederle Regina , Schnölzer Martina , Behrends Uta , Mautner Josef , Delecluse Henri-Jacques
Published: November 2009

The gp350 glycoprotein encoded by BLLF1 is crucial for efficient Epstein-Barr virus (EBV) infection of resting B cells. Gp350 binds to CD21, but whether this interaction sums up its functions remains unknown. We generated gp350-null EBVs that display CD19-, CD21-, or CD22-specific antibodies at their surface (designated as ΔBLLF1-Ab). Gp350-complemented (ΔBLLF1-C) and ΔBLLF1-Ab were found to bind equally well to B cells. Surprisingly, ΔBLLF1 binding was reduced only 1.7-fold relative to its complemented counterparts. Furthermore, B cells exposed to ΔBLLF1-Ab or ΔBLLF1 viruses presented structural antigens with comparable efficiency and achieved 25 to 80% of the T-cell activation elicited by ΔBLLF1-C. These findings show that the gp350-CD21 interaction pair plays only a modest role during virus transfer to the endosomal compartment. However, primary B cells or Raji B cells infected with ΔBLLF1-C viruses displayed a 35- to 70-fold higher infection rates than those exposed to ΔBLLF1, ΔBLLF1-CD22Ab, or ΔBLLF1-CD19Ab viruses. Complementation of the gp350 knockout phenotype with CD21Ab substantially enhanced infection rates relative to ΔBLLF1 but remained sevenfold (Raji B-cell line) to sixfold (primary B cells) less efficient than with gp350. We therefore infer that gp350 mainly exerts its functions after the internalization step, presumably during release of the viral capsid from the endosomal compartment, and that CD21-dependent but also CD21-independent molecular mechanisms are involved in this process. The latter appear to be characteristic of B-cell infection since transfection of CD21 in 293 cells improved the infection rates with both ΔBLLF1-CD21Ab and ΔBLLF1-C to a similar extent.
Musculoskeletal pain at multiple sites and its effects on work ability in a general working population
Authors: Miranda Helena , Kaila-Kangas Leena , Heliövaara Markku , Leino-Arjas Päivi , Haukka Eija , Liira Juha , Viikari-Juntura Eira
Published: November 2009

Objectives

Musculoskeletal pain often occurs at multiple sites concurrently. The aim of this study was to examine the associations between multi-site pain and self-rated work ability and retirement plans among actively working people.

Methods

The Health 2000 Survey was carried among a representative sample of Finnish adults. Musculoskeletal pain during the preceding month in the lower back, neck or shoulders, upper extremities, hips and lower extremities, and work ability and intentions to retire early were assessed. Subjects were also clinically examined. Analyses were restricted to 30–64-year-old subjects actively working during the preceding 12 months who provided information on work ability outcomes (population-weighted number of subjects=4087). Log-binomial regression was used to estimate prevalence ratios of reduced work ability.

Results

Single-site pain was reported by 33% of subjects, 20%, 9% and 4% reported pain in two, three and four sites, respectively, and 8%–15% reported poor work ability. Every fifth person had thought about retiring early. Age- and gender-adjusted risks of poor physical work ability and own prognosis of poor future work ability increased from 2 for single-site pain to 8 for pain at four sites. Risks remained considerably elevated after adjustment for various covariates, including clinical musculoskeletal disorders and functional capacity. Poor current work ability was most affected by multi-site pain at older age (50–64 years) and intentions to retire early at age 40–49 years.

Conclusions

Co-occurring pain is a considerable threat to work ability. Workers with multi-site pain may benefit from targeted preventive measures to sustain their work ability. Future studies should also consider multi-site pain as an important risk factor for reduced work ability.
Multimodality imaging in hyperparathyroidism
Authors: Patel C N , Scarsbrook A F
Published: November 2009

Imaging of patients with hyperparathyroidism has evolved over the last decade and transformed surgical management of the disease. Traditional bilateral neck dissection and exploratory surgery has been overtaken by focused, minimally invasive procedures in patients with single-gland disease. Accurate preoperative localisation of the involved parathyroid gland is the cornerstone for success of the new surgical techniques. In this article, the strengths, limitations and respective roles of different imaging modalities used to evaluate patients with hyperparathyroidism will be reviewed.
A doctor’s experience of resuscitation decision making for older patients: coping with change
Authors: Bowker L
Published: November 2009

There has been a dramatic change in the way that resuscitation decisions are made for older hospital inpatients in the UK. In less than 20 years many geriatric medicine wards have moved from an “opt-in” policy (where the default position is not to provide resuscitation) to a position where resuscitation is widely available to older patients and clinicians are much more reluctant to make “do not attempt cardiopulmonary resuscitation” decisions on medical grounds alone. The drivers for this change are rooted in societal changes rather than scientific or legal advances. Older patients are increasingly educated and have higher expectations of health care, while doctors are more commonly criticised and have progressively less “power” in the doctor–patient relationship. Doctors in the UK are struggling with the growing volume and complexity of guidelines about resuscitation, which can prove very tricky to enact within current resources and priorities. Many are feeling stressed and under equipped to cope with the ethical and legal dilemmas of resuscitation decision making. The solution lies in better undergraduate and postgraduate training which enhances skills (such as consultation skills and ethical reasoning) and not just knowledge of laws, principles and guidelines.
Citrus tissue culture: regulation of stylar abscission in excised pistils
Authors: Einset John W. , Cheng Anne , Elhag Hamid
Published: November 2009

Lemon pistil explants were obtained by cutting just above the region of the hypogynous disc (A type explant) or at the base of the pistil (B type explant) and cultured on test medium containing Murashige and Skoog salts, 50 g sucrose/L, 100 mg myo-inositol/L, 5 mg thiamine–HCl/L, and 0.5 mg kinetin/L, plus or minus supplements. Under appropriate conditions an abscission zone formed and styles abscised after 6–8 days of culture; in the field stylar abscission occurred 12–15 days postanthesis. Abscission in A type explants was markedly inhibited by 9 μM 2,4-dichlorophenoxyacetic acid but was unaffected by indole-3-acetic, 1-naphthaleneacetic, gibberellic, abscisic, caffeic, or p-coumaric acids. The response to 2,4-dichlorophenoxyacetic acid was reduced in B type explants. In an atmosphere containing 35–200 ppm ethylene, cell division occurred in the zone of stylar abscission producing a proliferating callus, and the content of cellulase increased from 0.6 to 53.7 enzyme units/g fresh weight compared with fresh explants. Stylar abscission was inhibited by 2,4-dichlorophenoxyacetic acid in A type explants of Washington navel orange, Valencia orange, and mandarin pistils, but not of grapefruit pistils. B type explants of Washington navel orange and mandarin pistils were less responsive to 2,4-dichlorophenoxyacetic acid.
Puya raimondii (Pitcairnioideae, Bromeliaceae) and birds: an hypothesis on nutrient relationships
Authors: Rees William E. , Roe Nicholas A.
Published: November 2009

We recorded observations on the relationships between Puya raimondii and the associated bird community in a dry grassland habitat in the high Andean plateau (altiplano or puna) of southeast Peru. From field evidence, we hypothesize that P. raimondii benefits from its association with birds through enhanced nutrient supply. Both bird droppings and dead birds trapped in the foliage of the plants represent a potential nutrient supplement. While the roots probably provide the principal uptake mechanism, the unique adaptations of other bromeliads respecting water and nutrient accumulation suggest that puyas may also have at least marginal capability for foliar absorption of water and mineral resources. The ecology of P. raimondii may illustrate the general conditions that favoured the independent evolution of specialized adaptations for xerophytism and epiphytism in other bromeliad taxa.
Patterns of species distribution in an Umbilicaria dominated community
Authors: Larson D. W.
Published: November 2009

Cliff and rock outcrop communities of which Umbilicaria lichens form a major component are described. In each of 210 sample sites in central Ontario, subjective estimates of species abundance were made together with measures of slope, aspect, and exposure. Analysis of the results shows that many species have specific requirements for slope and exposure. The significance of the observed patterns is discussed with respect to the known physiology of the Umbilicaria species.
An ultrastructural study of the marine diatom Licmophora hyalina and its parasite Ectrogella perforons. I. Infection of host cells
Authors: Kumar Chandralata Raghu
Published: November 2009

An electron microscopic study has been made on the infection and penetration of the marine diatom Licmophora hyalina Agardh by Ectrogella perforons Petersen, an obligate fungal parasite of diatoms. The zoospores encyst on the host cell wall. The nucleus of the cyst may be situated proximal or distal to the host wall. A germ tube is produced from the side where the nucleus is situated. The germ tube may be branched or unbranched. The penetrating germ tube swells distally, develops an appressorium at the site of penetration of the host wall, and pierces the host wall in the form of an infection peg. The infection peg is smaller in diameter than the germ tube and the appressorium. Successful infection takes place always at the areolae of the diatom wall. The infection peg may directly inject its contents by piercing the subfrustular layer of the diatom wall or may grow for some distance beneath the subfrustular layer. At the site of entry the host plasma membrane invaginates and surrounds the fungal protoplast. Initially, the host–parasite interface consists of a two-layered envelope of which the outer one is the host plasma membrane and the inner one the fungal plasma membrane.
New Therrya species parasitic on western conifers
Authors: Funk A.
Published: November 2009

Three new species of Therrya (Phacidiales) are described from diseased conifers in British Columbia, Canada, viz. T. pseudotsugae n.sp. on Pseudotsuga menziesii (Mirb.) Franco, T. tsugae n.sp. on Tsuga heterophylla (Raf.) Sarg., and T. piceae n.sp. on Picea glauca (Moench) Voss.
A histological and histochemical comparison of the mucilages on the root tips of several grasses
Authors: Miki N. K. , Clarke K. J. , McCully M. E.
Published: November 2009

Young, axenically grown roots of grasses are covered by two types of mucilage. Gelatinous material originates from the root cap, and a firm, uniformly thick mucilage overlies the columnar epidermal cells. Histochemical properties of these mucilages are similar in corn, wheat, barley, oats, sorghum, and a Sudan grass – sorghum hybrid.The epidermal mucilage has a thin outer and a thicker inner layer distinct from the epidermal cell wall. Both mucilage layers are strongly autofluorescent, birefringent, and PAS positive. Reactions of the outer layer and cell wall indicate carboxyl groups. These are absent from the inner mucilage. Root cap mucilage has a inner region with histochemical properties resembling those of the inner epidermal mucilage. The outer portion of the root cap mucilage is not fluorescent, not birefringent, weakly PAS positive, and carboxylated.
Penetration and growth of compatible and incompatible races of Phytophthora megasperma var. sojae in soybean hypocotyl tissues differing in age
Authors: Stössel P. , Lazarovits G. , Ward E. W. B.
Published: November 2009

Intact 6-day old soybean hypocotyls (Glycine max L., cv. Altona) were inoculated with zoospores of Phytophthora megasperma Drechs. var. sojae Hildeb. either at the top (susceptible to compatible races, resistant to incompatible races) or the bottom (resistant to both compatible and incompatible races) and, after a 22-h incubation, were examined by light microscopy. Penetration at the top and bottom by both compatible (race 6) and incompatible (race 4) P. megasperma var. sojae was predominantly between anticlinal walls of epidermal cells. Both races, but especially race 4, also penetrated directly into the outer walls of epidermal cells, but epidermal cells rarely were invaded. Both races grew mainly intercellularly, but race 6 produced haustoria more frequently than race 4. Race 6 haustoria at the top of the hypocotyl were usually encased, those of race 4 were not. Growth of both races was equally dense in the first few cell layers, but the numbers of race 4 hyphae decreased rapidly while those of race 6 became more abundant in the deeper layers. With race 4, but not with race 6, most cells in the infected tissue were necrotic. Differences between the compatible and the incompatible interactions were not absolute; there were many unsuccessful invasion attempts by race 6 and individual hyphae of race 4 spread deeply into the tissue. At the bottom of the hypocotyl, growth of both races was more restricted. Race 6 produced fewer haustoria than at the top, and similarities to the incompatible reaction with race 4 at the top suggest that similar mechanisms of resistance may be involved.
Influence of phenolics on in vitro growth of Frankia strains
Authors: Perradin Y. , Mottet M. J. , Lalonde M.
Published: November 2009

The influence of some common plant phenolics was tested on six Frankia strains isolated from both Alnus and Elaeagnus host plants. The addition of 1 mM of different phenolics to QmodB broth significantly influenced the growth and (or) the morphological development of Frankia. Ferulic, o-coumaric, and p-coumaric acids were extremely effective in inhibiting the growth of Frankia colonies, increasing the ramification of hyphae, and decreasing the number and size of sporangia produced in vitro. However, benzoic and p-hydroxybenzoic acids did not influence the total growth of Frankia colonies but stimulated the in vitro production of spherical septate vesicles on the two strains of Frankia type N tested. The possibility that these active phenolics, which are known to be present in Alnus actinorhizae, might act as "chemical mediators" between the host cell and its endophytic Frankia is suggested.
The isolation of Frankia from nodules of Casuarina
Authors: Diem H. G. , Dommergues Y.
Published: November 2009

Attempting to isolate Frankia from Casuarina, we found that this endophyte exhibited two unexpected characteristics: (i) an inability of most infected nodule cells to produce Frankia colonies; and (ii) the absence of specific nutritional requirements in the isolation medium. These findings led us to the formulation of some principles for isolating Frankia from Casuarina.
Physiology, chemistry, serology, and infectivity of two Frankia isolates from Alnus incana subsp. rugosa
Authors: Lechevalier Mary P. , Baker Dwight , Horrière Francoise
Published: November 2009

Actinorhizal nodules of Alnus incana ssp. rugosa collected from plants growing within 30 km of each other in Jeffersonville and Stowe, VT, U.S.A., yielded two morphologically distinct strains of Frankia. A comparison of the cell chemistry of the strains showed that each had the same type of cell wall and whole-cell sugar and phospholipid composition; however, AirI2 lacked the unknown amino acid found in the cells of all frankiae previously examined. Their cellular fatty acid composition showed only minor qualitative differences. Physiologically, the strains differed in their relation to oxygen, in their uptake and utilization of carbohydrates, organic acids, and lipids, and in protease production. AirI1 induced effective nodules in the host plant; those of AirI2 were ineffective. AirI2 was unrelated serologically to all frankiae previously isolated from the genus Alnus. AirI1 was serologically typical.
In vitro morphogenesis and ultrastructure of a Frankia sp. ArI3 (Actinomycetales) from Alnus rubra and a morphologically similar isolate (AirI2) from Alnus incana subsp. rugosa
Authors: Horrière Françoise , Lechevalier Mary P. , Lechevalier Hubert A.
Published: November 2009

An actinomycete (AirI2) with black spores was isolated from nodules of Alnus incana ssp. rugosa. Although this organism is very different in appearance from the typically white strains of Frankia from Alnus, it has all the main microscopic features of members of this genus. However, cytological differences have been observed when compared with a strain with white spores (ArI3).
Deoxyribonucleic acid base composition of 12 Frankia isolates
Authors: An Chung Sun , Wills John W. , Riggsby W. Stuart , Mullin Beth C.
Published: November 2009

The base compositions of the DNA from 12 Frankia isolates were determined. The guanine plus cytosine content fell within the range of 68.4–72.1 mol% for all of the isolates tested. The DNA's from one Alnus isolate, one Comptonia isolate, and two Elaeagnus isolates were analyzed for the occurrence of modified bases, and none were found.
Effects of Pseudomonas cepacia and cultural factors on the nodulation of Alnus rubra roots by Frankia
Authors: Knowlton Susan , Dawson Jeffrey O.
Published: November 2009

Infection of Alnus rubra Bong, roots by Frankia isolates was consistently promoted by Pseudomonas cepacia "helper" bacteria under a variety of cultural conditions. Pseudomonas cepacia, while helpful, was never necessary in securing nodulation of aseptic A. rubra by Frankia. Live P. cepacia cells were added with a Frankia isolate to aseptically grown A. rubra seedlings on Hoagland's agar slants. This doubled, on average, the number of nodules formed by Frankia alone when Frankia isolate, age of Frankia inoculum, Frankia medium, calcium concentration of seedling substrate, phosphate concentration of seedling substrate, and pH of seedling substrate were varied. There was an apparent interaction between seedling substrate pH and P. cepacia in the promotion of nodulation. At pH levels from 4.0 to 6.0 the infectivity of Frankia isolate ArI3 alone was greatly depressed compared with infectivity at the optimal pH for the growth of ArI3, around 7.0. However, at pH levels of 5.0, 5.5, and 6.0 the ability of P. cepacia to promote nodulation of A. rubra seedlings by ArI3 was greater than at pH 7.0. This interaction may be due to the ability of P. cepacia to neutralize and grow more rapidly in mildly acidic solutions in our experimental system, combined with the ability of P. cepacia to cause root-hair deformation, which has been associated with the actinorhizal infection process.