bluuebunny/crossref_metadata_2025 huggingface dataset:
Comparison of Hemoglobin A1c and Fasting Glucose Criteria to Diagnose Diabetes Among People With Metabolic Syndrome and Fasting Glucose Above 100 mg/dL (5.5 mmol/L)
Authors: Liberopoulos Evangelos N. , Florentin Matilda , Kei Anastazia , Mountzouri Elisavet , Agouridis Aris , Elisaf Moses S.
Published: May 2010

J Clin Hypertens(Greenwich). 2010;12:543–548. © 2010 Wiley Periodicals, Inc.

The aim of this study was to compare hemoglobin A1c (HbA1c) and fasting glucose for the diagnosis of diabetes among people with metabolic syndrome and fasting glucose >100 mg/dL (5.5 mmol/L). Consecutive individuals (N=142) with metabolic syndrome and fasting glucose >100 mg/dL (5.5 mmol/L) but without a self‐reported history of diabetes who visited the outpatient lipid and obesity clinic of the University Hospital of Ioannina, Greece from January through September 2009 were included. HbA1c≥6.5% and fasting glucose ≥126 mg/dL (7 mmol/L) were used separately to define diabetes. Overall, 29.5% of patients had both HbA1c≥6.5% and fasting glucose ≥126 mg/dL (7 mmol/L), 25.3% had HbA1c≥6.5% but fasting glucose <126 mg/dL (7 mmol/L), and 9.1% had HbA1c <6.5% but fasting glucose ≥126 mg/dL (7 mmol/L). A greater proportion of patients reached a diagnosis of diabetes based on the HbA1c criterion (n=78, 54.9%) compared with the fasting glucose criterion (n=55, 38.7%, P=.000). A large proportion of patients (44.8%) with impaired fasting glucose (fasting glucose 100–125 mg/dL; 5.6–6.9 mmol/L) would be classified as diabetics using the HbA1c criterion. Implication of the HbA1c criterion may increase the rate of diabetes diagnosis among people with metabolic syndrome and fasting glucose >100 mg/dL (5.5 mmol/L).
A Novel Echocardiographic Marker in Hypertensive Patients: Is Diastolic Dysfunction Associated With Atrial Electromechanical Abnormalities in Hypertension?
Authors: Yavuz Bunyamin , Deniz Ali , Ertugrul Derun Taner , Deveci Onur Sinan , Yalcin Ahmet Arif , Ata Naim , Kucukazman Metin , Dal Kursat , Tutal Emre
Published: May 2010

J Clin Hypertens (Greenwich). 2010;12:687–692. ©2010 Wiley Periodicals, Inc.

Atrial arrhythmias are common problems in hypertensive patients. Atrial electromechanical delay (AEMD) can be used to evaluate development of atrial arrhythmias. The authors aimed to assess inter‐ and intra‐AEMD in hypertensive patients. The study population consisted of 200 medically treated hypertensive patients and 151 normotensive controls. Inter‐AEMD and intra‐left AEMD were measured from parameters of Doppler tissue imaging. There were 72 (36%) hypertensive patients with diastolic dysfunction, 128 (64%) patients without diastolic dysfunction, and 151 healthy controls. Inter‐AEMD (59 ms [36–104 ms] vs 42 ms [36–68 ms] vs 46 ms [30–82 ms]) was significantly higher in hypertensive patients with diastolic dysfunction compared with patients without diastolic dysfunction and controls. Our data demonstrated that inter‐AEMD is longer in hypertensive patients with diastolic dysfunction. It may be suggested that diastolic dysfunction is associated with atrial electromechanical abnormalities, which can be associated with atrial fibrillation in hypertension.
Problematic Within‐Family Alliances in Conjoint Family Therapy: A Close Look at Five Cases
Authors: Lambert Jessica E. , Skinner Alyson H. , Friedlander Micki L.
Published: May 2010

We intensively analyzed five sessions in which the within‐family alliance was manifestly problematic and family members reported discrepant perceptions of the family’s sense of purpose about working together in therapy. Triangulation of observer ratings with (a) session content, (b) participants’ postsession perceptions, (c) therapists’ alliance‐related interventions, and (d) clients’ pretreatment target complaints and problem descriptions suggested three essential, interrelated aspects of the within‐family alliance: family members (a) agreeing on the problem(s) and goals for treatment, (b) feeling connected in coping with their concerns (“a felt unity”), and (c) seeing conjoint therapy as valuable. Within‐family disagreements on any of these aspects seemed to account for the observed in‐session struggles. The analysis also suggested several missed opportunities for strengthening the within‐family alliance.
Using Mindfulness Meditation to Teach Beginning Therapists Therapeutic Presence: A Qualitative Study
Authors: McCollum Eric E. , Gehart Diane R.
Published: May 2010

Some of the more difficult to define aspects of the therapeutic process (empathy, compassion, presence) remain some of the most important. Teaching them presents a challenge for therapist trainees and educators alike. In this study, we examine our beginning practicum students’ experience of learning mindfulness meditation as a way to help them develop therapeutic presence. Through thematic analysis of their journal entries a variety of themes emerged, including the effects of meditation practice, the ability to be present, balancing being and doing modes in therapy, and the development of acceptance and compassion for themselves and for their clients. Our findings suggest that mindfulness meditation may be a useful addition to clinical training.
Integrated river basin management in Southeast Asia
Authors: Meigh J.D. , Bartlett J.M.
Published: May 2010

The impacts and aftermath of floods affect millions of people worldwide and the poor in particular. In recent years, there has been a worldwide change in approach from flood control to flood management. The former relies mainly on engineering interventions and often achieves only localised protection. In the latter, a more holistic approach is adopted, where the whole river basin is treated as an integrated system. This paper examines three river basins in Southeast Asia where policies generated by the World Bank and Asian Development Bank were applied to determine the critical causes of flooding and the main interfaces with other water sectors.
Dystrophia Helsinglandica – corneal morphology, topography and sensitivity in a hereditary corneal disease with recurrent erosive episodes
Authors: Neira Waldir , Hammar Björn , Holopainen Juha M. , Tuisku Ilpo , Dellby Anette , Tervo Timo , Fagerholm Per
Published: May 2010

Purpose:  The aim of this study was to describe the morphology, corneal topography and sensitivity in individuals with Dystrophia Helsinglandica. This autosomal dominant corneal disease is characterized by recurrent corneal erosive episodes and progressive subepithelial fibrosis not significantly affecting visual acuity.

Methods:  The corneas of nine affected and nine unaffected individuals were examined using slit‐lamp biomicroscopy, in vivo confocal microscopy (IVCM) and videokeratography. Corneal mechanical sensitivity was also measured using a non‐contact esthesiometer.

Results:  Slit‐lamp biomicroscopy revealed that the affected individuals represented different stages of corneal changes, from a nearly normal cornea to subepithelial fibrosis of the central cornea. Corneal changes in affected individuals did not significantly decrease the best spectacle‐corrected visual acuity. In vivo confocal microscopy detected morphological changes in the epithelium and stroma. Subepithelial opacity formation including altered keratocytes could be found in the anterior stroma in all affected eyes. With the exception of two eyes (one affected and one unaffected), all videokeratographies showed irregular astigmatism. Corneal sensitivity was significantly lower in affected individuals (p = 0.01). Age and corneal sensitivity showed no correlation.

Conclusion:  The main morphological findings in affected individuals were discrete and progressive subepithelial fibrosis, in the in vivo confocal microscope corresponding to optically dense extracellular matrix and activated keratocytes. Subbasal nerve morphology was changed in the affected family members who also showed a decreased corneal sensitivity. The findings are per se not specific to the disease. The changes probably reflect a healing response to erosive events on the corneal surface influenced by the genotype.
Outcome of capsulotomy in patients with low vision and posterior capsule opacification
Authors: Sundelin Karin , Lundström Mats
Published: May 2010

Purpose:  To investigate the functional outcome, as defined using the Catquest questionnaire, for patients with posterior capsule opacification (PCO) and low corrected distance visual acuity (CDVA), after capsulotomy.

Methods:  Thirty‐one patients with PCO and visual acuity ≥0.6 (logMAR, ≤0.25 decimal notation) in the PCO eye were examined before and 1 month after capsulotomy. An ophthalmic examination, procuring of a digital image of the posterior lens capsule and completion of Catquest were carried out.

Results:  Mean CDVA (p < 0.001), disabilities in daily life (p = 0.004), activity (p = 0.012), symptoms of glare (p = 0.003) and satisfaction with vision (p < 0.001) all improved after capsulotomy. Sixty‐five per cent of patients had good or very good benefit and 23% questionable or no benefit. The group with moderate benefit was only 6%. Compared with patients with PCO and better visual acuity, the number of patients with good benefit was greater than the number with moderate benefit.

Conclusions:  Most patients with PCO and low CDVA reported improved visual function in daily life as defined using Catquest, after capsulotomy. The group with moderate benefit was small, indicating that the improvement was either substantial or negligible.
One‐stage explant–implant procedure of exposed porous orbital implants
Authors: Toft Peter B. , Roed Rasmussen Marie L. , Prause Jan U.
Published: May 2010

Purpose:  To investigate the risks of implant exposure after a combined explant–implant procedure in patients with an exposed porous orbital implant.

Methods:  Twenty‐four consecutive patients who had a combined explant–implant procedure of an exposed hydroxyapatite (21) or porous polyethylene (3) orbital implant from January 2000 to February 2009 were included. The patient records were reviewed; patients were interviewed by telephone and invited for a clinical examination. Histopathological examination was carried out on the removed implants. Main outcome measures were: presence of exposure of the new implant or not, patient graded satisfaction with the cosmetic result, and presence of poor motility.

Results:  None of the new implants became exposed or infected in the follow‐up period of 25 [3–94] months (median [range]). The patients scored their satisfaction with the cosmetic result to a median score of 9 (range 5–10). Poor motility was present in six of 17 patients. Micro‐organisms were identified in three removed implants and signs of inflammation were present in 20 removed implants.

Conclusions:  If a decision of implant removal has been made, it is safe to replace the implant at the same procedure in sockets without profound signs of infection. The procedure carries a possible risk of poor motility.
The effect of Kashin‐Beck disease‐affected feed and T‐2 toxin on the bone development of Wistar rats
Authors: YAN Denglu , KANG Pengde , YANG Jing , SHEN Bing , ZHOU Zongke , DUAN Lijun , DENG Jiayun , HUANG Hui , PEI Fu‐Xing
Published: May 2010

Objective:  To investigate the effect of Kashin‐Beck disease (KBD)‐affected feed and T‐2 toxin on the bone development of Wistar rats.

Methods:  Seventy‐eight ablactation Wistal rats (50% male and 50% female) weighing ≈ 65 g were obtained from Sichuan Medical Center (Chengdu, China), and were randomly assigned to three groups (Groups A, B and C). Group A had 24 rats and were fed with commercial rat feed (control); Group B had 30 rats and were fed with commercial rat feed and T2 toxin by intragastric administration; and Group C had 24 rats and were fed with the KBD‐affected feed. The histological sections were stained with hematoxylin and eosin (H&E) and Masson dye.

Results:  Weight gain was fastest Group A rats and Group C rats had the lowest weight gain (P < 0.05). There were no epiphyseal plate chondrocyte necroses in the control group at the first, second, and fourth weeks. In the T‐2 toxin group, two rats had chondrocyte‐focus necroses at the labrocyte cell zone at the second week. At the fourth week, six rats had chondrocyte‐focus or lamellar necroses at the labrocyte cell zone. Three rats had focus necrosis at the proliferation cell zone, and there were three rats with penetration necrosis. In the KBD‐affected group, one rat had chondrocyte‐focus necrosis at the labrocyte cell zone at the second week and seven rats had chondrocyte‐focus necrosis at the labrocyte cell zone at the fourth week. And at the same time, two rats had focus necrosis at the proliferation cell zone, three rats had lamellar necrosis at the labrocyte cell zone, four had focus necrosis at the labrocyte cell zone, and two rats had penetration necrosis. The epiphyseal plate Masson dye of the control group showed deep blue collogen coloration and in the KBD‐affected group and T‐2 toxin group, collogen showed a pale blue color, the drum dyeing was uneven, and the collogen was showed an absence of color in the region of the necrosis.

Conclusions:  With KBD‐affected feed or T‐2 toxin intervention, rats had focus necrosis and lamellar necrosis at the epiphyseal plate. KBD‐affected feed rats had less weight gain than T‐2 toxin intervention rats, which means there were other etiological factors in KBD‐affected feed.
Correlation between structural and optical properties of a-plane GaN films grown on r-plane sapphire by metal organic chemical-vapor deposition
Authors: Jung Mina , Chang Jiho , Lee Hyunjae , Ha Jun-seok , Park Jin-sub , Park Seungwhan , Fujii Katsushi , Yao Takafumi , Kil Gyung-suk , Lee Seogwoo , Cho Myungwhan , Whang Sungmin , Seo Yong-gon
Published: May 2010

The authors have studied the photoluminescence (PL) intensities of a-plane (11-20) GaN films (a-GaN) as a function of x-ray rocking-curve (XRC) linewidth values measured in both c- and m-axis directions. PL intensity of well-known luminescence lines such as 3.47 eV (bound exciton emission), 3.41 eV (basal-plane stacking fault related emission), and 3.29 eV (defect induced emission) are discussed in terms of XRC linewidth values. PL intensities reveal a close relationship with XRC linewidth measured in the c-axis direction, while an unusual relationship was observed between PL intensity and XRC linewidth in the m-axis direction. Inhomogeneous strain along the m-axis direction of a-GaN film is discussed as a cause of XRC linewidth broadening, rather than the formation of structural defects.
Simulation of self-focusing electron emitter
Authors: Yuan Guang , Jiang Jinjing , Li Chun , Liu Weidong , Mimura Hitenori
Published: May 2010

A new structure of field electron emitter was proposed to reduce the dispersion of electron beam emitted. In this structure the gate electrode is placed at the center and surrounded by cathode electrode, and then the gate electrode not only plays extracting electrons from cathode but also focusing the electrons emitted. The distribution of the electric field and the electron trajectory of the emitter were investigated and the optimum parameters including geometrical and electrical parameters about the electron emitter are obtained.
Response to Comment on the Paleobiology and Classification of
Ardipithecus ramidus
Authors: White Tim D. , Suwa Gen , Lovejoy C. Owen
Published: May 2010

We assigned
Ardipithecus
to the Hominidae based on numerous dental, cranial, and postcranial characters. Sarmiento argues that these characters are not exclusive to hominids, contending that
Ardipithecus
is too old to be cladistically hominid. His alternative phylogeny, however, is unlikely because it requires tortuous, nonparsimonious evolutionary pathways.
Fairness and the Development of Inequality Acceptance
Authors: Almås Ingvild , Cappelen Alexander W. , Sørensen Erik Ø. , Tungodden Bertil
Published: May 2010

Fairness or Equality?

Inequality in payments may be seen as inherently unfair, or as appropriate when it reflects differential achievement. Using an economic exchange game,

Almås
et al.

(p.
1176
) mapped how judgments changed from 5th-grade students to 13th graders: Fifth graders expressed a preference for equal division of rewards, whereas the 13th graders tolerated unequal outcomes, as long as they had been provided with evidence of unequal inputs. That is, the younger children were strict egalitarians, but the older ones—perhaps as a consequence of exposure to a variety of achievement-based social activities, such as sports—tended toward meritocracy.
Self-Assembled Plasmonic Nanoparticle Clusters
Authors: Fan Jonathan A. , Wu Chihhui , Bao Kui , Bao Jiming , Bardhan Rizia , Halas Naomi J. , Manoharan Vinothan N. , Nordlander Peter , Shvets Gennady , Capasso Federico
Published: May 2010

Optical Nanoengineering

Optics and electronics operate at very different length scales. Surface plasmons are light-induced electronic excitations that are being pursued as a route to bridge the length scales and bring the processing speed offered by optical communication down to the size scales of electronic chip circuitry. Now,

Fan
et al.

(p.
1135
) describe the self-assembly of nanoscale dielectric particles coated with gold. Functionalization of the gold surface with polymer ligands allowed controlled production of clusters of nanoparticles. The optical properties of the self-assembled nanostructures depended on the number of components within the cluster and each structure could be selected for its unique optical properties. Such a bottom-up approach should help in fabricating designed optical circuits on the nanoscale.
Nitrogen in the Solar System
Authors: Marti Kurt , Kerridge John
Published: May 2010

Nitrogen isotopic compositions can serve as useful tracers to understand the input materials and processes during formation and evolution of the solar system.
Prion Strain Mutation and Selection
Authors: Collinge John
Published: May 2010

Structural compatibility of infecting prion proteins with those of a new host determine whether they will be successfully transmitted.
Efforts to Stop UNESCO Science Award in Honor of African Dictator
Authors: Enserink Martin
Published: May 2010

Unless human-rights organizations score a last-minute victory, one or more life scientists could find themselves in an awkward spot next month: to be awarded a prize by the United Nations Educational, Scientific and Cultural Organization (UNESCO) that is named after one of Africa's most infamous dictators.
Francis Collins: On Recruiting Varmus, Discovering Drugs, the Funding Cliff
Authors: Kaiser Jocelyn
Published: May 2010

Last week in an interview with several
Science
staffers, National Institutes of Health Director Francis Collins discussed the budget prospects of the for next year and spoke of his role in luring former NIH director Harold Varmus back to head the National Cancer Institute.
Lowering the Boom on Financial Conflicts
Authors: Kaiser Jocelyn
Published: May 2010

After a year of review, the National Institutes of Health has unveiled a plan for cracking down on conflicts of interest in biomedical research. The changes would require researchers to report more financial information to their employers, have the institutions and not researchers decide when a conflict exists, and make some of the reported information public.
Exploring the Virgin Country
Authors: Lawler Andrew
Published: May 2010

Omani soil is providing a bevy of archaeological surprises, but researchers there also struggle with rapid development and a lack of homegrown expertise.
Evaluation of the Analytical Performance of the Xpert MTB/RIF Assay
Authors: Blakemore Robert , Story Elizabeth , Helb Danica , Kop JoAnn , Banada Padmapriya , Owens Michelle R. , Chakravorty Soumitesh , Jones Martin , Alland David
Published: May 2010

We performed the first studies of analytic sensitivity, analytic specificity, and dynamic range for the new Xpert MTB/RIF assay, a nucleic acid amplification-based diagnostic system that detects
Mycobacterium tuberculosis
and rifampin (RIF) resistance in under 2 h. The sensitivity of the assay was tested with 79 phylogenetically and geographically diverse
M. tuberculosis
isolates, including 42 drug-susceptible isolates and 37 RIF-resistant isolates containing 13 different
rpoB
mutations or mutation combinations. The specificity of the assay was tested with 89 nontuberculosis bacteria, fungi, and viruses. The Xpert MTB/RIF assay correctly identified all 79
M. tuberculosis
isolates and correctly excluded all 89 nontuberculosis isolates. RIF resistance was correctly identified in all 37 resistant isolates and in none of the 42 susceptible isolates. Dynamic range was assessed by adding 10
2
to 10
7
CFU of
M. tuberculosis
into
M. tuberculosis
-negative sputum samples. The assay showed a log-linear relationship between cycle threshold and input CFU over the entire concentration range. Resistance detection in the presence of different mixtures of RIF-resistant and RIF-susceptible DNA was assessed. Resistance detection was dependent on the particular mutation and required between 65% and 100% mutant DNA to be present in the sample for 95% certainty of resistance detection. Finally, we studied whether assay specificity could be affected by cross-contaminating amplicons generated by the GenoType MTBDRplus assay.
M. tuberculosis
was not detected until at least 10
8
copies of an MTBDRplus amplicon were spiked into 1 ml of sputum, suggesting that false-positive results would be unlikely to occur.
Rapid Multiple-Locus Variant-Repeat Assay (MLVA) for Genotyping of
Streptococcus agalactiae
Authors: Radtke Andreas , Lindstedt Bjørn-Arne , Afset Jan Egil , Bergh Kåre
Published: May 2010

Several methods have been used for typing of
Streptococcus agalactiae
(group B streptococci [GBS]). Methods currently in use may provide inadequate resolution (e.g., typing of capsular polysaccharides and surface protein) or are labor-intensive and expensive (e.g., multilocus sequence typing [MLST] or pulsed-field gel electrophoresis). This work describes the construction and use of a multiple-locus variant-repeat assay (MLVA) on 126 well-characterized human GBS strains, consisting mostly of invasive Norwegian strains and international reference strains. Based on
in silico
whole-genomic analysis of the genomes of strains A909, NEM316, and 2603V/R, 18 candidate loci were selected and investigated by PCR. Eleven loci showed diversity, and the five most diverse loci were used for the construction of an MLVA, consisting of a multiplex PCR followed by fragment analysis with capillary electrophoresis. The assay generated clusters which corresponded well with those observed by other methods. However, it provided a considerably higher degree of diversity, with 70 different MLVA types compared to 36 types generated by MLST. Simpson's index of diversity for the 5-locus MLVA was 0.963, compared to 0.899 for the MLST in this strain collection. MLVA results will generally be available within 2 days, which is usually faster than MLST. In our hands, MLVA of GBS represents a rapid, easy, and comparably inexpensive method for high-resolution genotyping of GBS.
Streptococcus salivarius
Meningitis Case Strain Traced to Oral Flora of Anesthesiologist
Authors: Shewmaker Patricia L. , Gertz Robert E. , Kim Clara Y. , de Fijter Sietske , DiOrio Mary , Moore Matthew R. , Beall Bernard W.
Published: May 2010

Two women in labor received intrapartum spinal anesthesia from the same anesthesiologist approximately 1 h apart. Within 15 h, both patients developed
Streptococcus salivarius
meningitis and one patient died. Blood and cerebrospinal fluid (CSF) samples from both patients and tongue swab specimens from the anesthesiologist yielded isolates of an indistinguishable
S. salivarius
strain.
Characteristics of Meropenem Heteroresistance in
Klebsiella pneumoniae
Carbapenemase (KPC)-Producing Clinical Isolates of
K. pneumoniae
Authors: Pournaras Spyros , Kristo Ioulia , Vrioni Georgia , Ikonomidis Alexandros , Poulou Aggeliki , Petropoulou Dimitra , Tsakris Athanassios
Published: May 2010

Meropenem heteroresistance was investigated in six apparently meropenem-susceptible,
Klebsiella pneumoniae
carbapenemase (KPC)-producing
K. pneumoniae
(KPC-KP) clinical isolates, compared with that in carbapenemase-negative, meropenem-susceptible controls. In population analyses, the KPC-KP isolates grew at meropenem concentrations of 64 to 256 μg/ml. Heteroresistant colonies had significantly elevated expression of the
bla
KPC
gene compared with the native populations but did not retain heteroresistance when subcultured in drug-free media. Time-kill assays indicated that meropenem alone was not bactericidal against KPC-KP but efficiently killed the control strains.
Establishment and Characterization of a Madin-Darby Canine Kidney Reporter Cell Line for Influenza A Virus Assays
Authors: Hossain M. Jaber , Perez Sandra , Guo Zhu , Chen Li-Mei , Donis Ruben O.
Published: May 2010

Influenza virus diagnosis has traditionally relied on virus isolation in chicken embryo or cell cultures. Many laboratories have adopted rapid molecular methods for detection of influenza viruses and discontinued routine utilization of the relatively slow viral culture methods. We describe an influenza A virus reporter cell line that contributes to more efficient viral detection in cell culture. Madin-Darby canine kidney (MDCK) cells were engineered to constitutively produce an influenza virus genome-like luciferase reporter RNA driven by the canine RNA polymerase I promoter. Induction of a high level of luciferase activity was detected in the Luc9.1 cells upon infection with various strains of influenza A virus, including 2009 H1N1 pandemic and highly pathogenic H5N1 virus. In contrast, infection with influenza B virus or human adenovirus type 5 did not induce significant levels of reporter expression. The reporter Luc9.1 cells were evaluated in neutralizing antibody assays with convalescent H3N2 ferret serum, yielding a neutralization titer comparable to that obtained by the conventional microneutralization assay, suggesting that the use of the reporter cell line might simplify neutralization assays by facilitating the establishment of infectious virus endpoints. Luc9.1 cells were also used to determine the susceptibility of influenza A viruses to a model antiviral drug. The equivalence to conventional antiviral assay results indicated that the Luc9.1 cells could provide an alternative cell-based platform for high-throughput drug discovery screens. In summary, the MDCK-derived Luc9.1 reporter cell line is highly permissive for influenza A virus replication and provides a very specific and sensitive approach for simultaneous detection and isolation of influenza A viruses as well as functional evaluation of antibodies and antiviral molecules.
Human Cytomegalovirus pUL83 Stimulates Activity of the Viral Immediate-Early Promoter through Its Interaction with the Cellular IFI16 Protein
Authors: Cristea Ileana M. , Moorman Nathaniel J. , Terhune Scott S. , Cuevas Christian D. , O'Keefe Erin S. , Rout Michael P. , Chait Brian T. , Shenk Thomas
Published: May 2010

The human cytomegalovirus (HCMV) virion protein pUL83 (also termed pp65) inhibits the expression of interferon-inducible cellular genes. In this work we demonstrate that pUL83 is also important for efficient induction of transcription from the viral major immediate-early promoter. Infection with a mutant virus containing a premature translation termination codon in the UL83 open reading frame (ORF) (UL83Stop) resulted in decreased transcription from the major immediate-early promoter in a time- and multiplicity-dependent manner. Expression of pUL83 alone is capable of transactivating the promoter in a reporter assay, and pUL83 associates with the promoter in infected cells. To investigate the mechanism by which the protein regulates the major immediate-early promoter, we utilized a mutant virus expressing an epitope-tagged pUL83 from its own promoter to identify protein binding partners for pUL83 during infection. We identified and confirmed the interaction of pUL83 with cellular IFI16 family members throughout the course of HCMV infection. pUL83 recruits IFI16 to the major immediate-early promoter, and IFI16 binding at the promoter is dependent upon the presence of pUL83. Consistent with the results obtained with the UL83Stop virus, infection of IFI16 knockdown cells with wild-type virus resulted in decreased levels of immediate-early transcripts compared to those of control cells. These data identify a previously unknown role for pUL83 in the initiation of the human cytomegalovirus gene expression cascade.
Induction of Neutralizing Antibody Responses to Anthrax Protective Antigen by Using Influenza Virus Vectors: Implications for Disparate Immune System Priming Pathways
Authors: Langley William A. , Bradley Konrad C. , Li Zhu-Nan , Smith Mary Ellen , Schnell Matthias J. , Steinhauer David A.
Published: May 2010

Viral vectors based on influenza virus, rabies virus (RV), and vaccinia virus (VV) were used to express large polypeptide segments derived from the
Bacillus anthracis
protective antigen (PA). For the infectious influenza virus vector and recombinant VV constructs, the receptor binding domain (RBD or domain 4) or the lethal and edema factor binding domain (LEF or domain 1′) were engineered into functional chimeric hemagglutinin (HA) glycoproteins. In the case of the RV vector, the viral glycoprotein (G) was used as a carrier for RBD in an inactivated form of the vector. These constructs were examined by using multiple homologous and heterologous prime/boost immunization regimens in order to optimize the induction of α-PA antibody responses. Several immunization combinations were shown to induce high titers of antibody recognizing the anthrax RBD and LEF domains, as well as the full-length PA protein in mice. The heterologous prime/boost immunization regimens that involved an initial intranasal administration of a live influenza virus vector, followed by an intramuscular boost with either the killed RV vector or the VV vector, were particularly effective, inducing antigen-specific antibodies at levels severalfold higher than homologous or alternative heterologous protocols. Furthermore, sera from several groups of the immunized mice demonstrated neutralization activity in an
in vitro
anthrax toxin neutralization assay. In some cases, such toxin-neutralizing activity was notably high, indicating that the mechanisms by which immunity is primed by live influenza virus vectors may have beneficial properties.
The Cysteine Protease Domain of Porcine Reproductive and Respiratory Syndrome Virus Nonstructural Protein 2 Possesses Deubiquitinating and Interferon Antagonism Functions
Authors: Sun Zhi , Chen Zhenhai , Lawson Steven R. , Fang Ying
Published: May 2010

Porcine reproductive and respiratory syndrome (PRRS) virus nonstructural protein 2 (nsp2) contains a cysteine protease domain at its N terminus, which belongs to the ovarian tumor (OTU) protease family. In this study, we demonstrated that the PRRSV nsp2 OTU domain antagonizes the type I interferon induction by interfering with the NF-κB signaling pathway. Further analysis revealed that the nsp2 OTU domain possesses ubiquitin-deconjugating activity. This domain has the ability to inhibit NF-κB activation by interfering with the polyubiquitination process of IκBα, which subsequently prevents IκBα degradation. To determine whether the nsp2 protein antagonist function can be ablated from the virus, we introduced point mutations into the OTU domain region by use of reverse genetics. The D458A, S462A, and D465A mutations targeting on a B-cell epitope in the OTU domain region generated the viable recombinant viruses, and the S462A and D465A mutants were attenuated for growth in cell culture. The OTU domain mutants were examined to determine whether mutations in the nsp2 OTU domain region altered virus ability to inhibit NF-κB activation. The result showed that certain mutations lethal to virus replication impaired the ability of nsp2 to inhibit NF-κB activation but that the viable recombinant viruses, vSD-S462A and vSD-D465A, were unable to inhibit NF-κB activation as effectively as the wild-type virus. This study represents a fundamental step in elucidating the role of nsp2 in PRRS pathogenesis and provides an important insight in future modified live-virus vaccine development.
Respiratory Syncytial Virus Engineered To Express the Cystic Fibrosis Transmembrane Conductance Regulator Corrects the Bioelectric Phenotype of Human Cystic Fibrosis Airway Epithelium
In Vitro
Authors: Kwilas Anna R. , Yednak Mark A. , Zhang Liqun , Liesman Rachael , Collins Peter L. , Pickles Raymond J. , Peeples Mark E.
Published: May 2010

Cystic fibrosis (CF) is the most common lethal recessive genetic disease in the Caucasian population. It is caused by mutations in the CF transmembrane conductance regulator (CFTR) gene that is normally expressed in ciliated airway epithelial cells and the submucosal glands of the lung. Since the CFTR gene was first characterized in 1989, a major goal has been to develop an effective gene therapy for CF lung disease, which has the potential to ameliorate morbidity and mortality. Respiratory syncytial virus (RSV) naturally infects the ciliated cells in the human airway epithelium. In addition, the immune response mounted against an RSV infection does not prevent subsequent infections, suggesting that an RSV-based vector might be effectively readministered. To test whether the large 4.5-kb CFTR gene could be expressed by a recombinant RSV and whether infectious virus could be used to deliver CFTR to ciliated airway epithelium derived from CF patients, we inserted the CFTR gene into four sites in a recombinant green fluorescent protein-expressing RSV (rgRSV) genome to generate virus expressing four different levels of CFTR protein. Two of these four rgRSV-CFTR vectors were capable of expressing CFTR with little effect on viral replication. rgRSV-CFTR infection of primary human airway epithelial cultures derived from CF patients resulted in expression of CFTR protein that was properly localized at the luminal surface and corrected the chloride ion channel defect in these cells.
Evaluation of Replication and Cross-Reactive Antibody Responses of H2 Subtype Influenza Viruses in Mice and Ferrets
Authors: Chen Grace L. , Lamirande Elaine W. , Yang Chin-Fen , Jin Hong , Kemble George , Subbarao Kanta
Published: May 2010

H2 influenza viruses have not circulated in humans since 1968, and therefore a large segment of the population would likely be susceptible to infection should H2 influenza viruses reemerge. The development of an H2 pandemic influenza virus vaccine candidate should therefore be considered a priority in pandemic influenza preparedness planning. We selected a group of geographically and temporally diverse wild-type H2 influenza viruses and evaluated the kinetics of replication and compared the ability of these viruses to induce a broadly cross-reactive antibody response in mice and ferrets. In both mice and ferrets, A/Japan/305/1957 (H2N2), A/mallard/NY/1978 (H2N2), and A/swine/MO/2006 (H2N3) elicited the broadest cross-reactive antibody responses against heterologous H2 influenza viruses as measured by hemagglutination inhibition and microneutralization assays. These data suggested that these three viruses may be suitable candidates for development as live attenuated H2 pandemic influenza virus vaccines.
African Swine Fever Virus Protein p17 Is Essential for the Progression of Viral Membrane Precursors toward Icosahedral Intermediates
Authors: Suárez Cristina , Gutiérrez-Berzal Javier , Andrés Germán , Salas María L. , Rodríguez Javier M.
Published: May 2010

The first morphological evidence of African swine fever virus (ASFV) assembly is the appearance of precursor viral membranes, thought to derive from the endoplasmic reticulum, within the assembly sites. We have shown previously that protein p54, a viral structural integral membrane protein, is essential for the generation of the viral precursor membranes. In this report, we study the role of protein p17, an abundant transmembrane protein localized at the viral internal envelope, in these processes. Using an inducible virus for this protein, we show that p17 is essential for virus viability and that its repression blocks the proteolytic processing of polyproteins pp220 and pp62. Electron microscopy analyses demonstrate that when the infection occurs under restrictive conditions, viral morphogenesis is blocked at an early stage, immediately posterior to the formation of the viral precursor membranes, indicating that protein p17 is required to allow their progression toward icosahedral particles. Thus, the absence of this protein leads to an accumulation of these precursors and to the delocalization of the major components of the capsid and core shell domains. The study of ultrathin serial sections from cells infected with BA71V or the inducible virus under permissive conditions revealed the presence of large helicoidal structures from which immature particles are produced, suggesting that these helicoidal structures represent a previously undetected viral intermediate.
Differential Effects of the Putative GBF1 Inhibitors Golgicide A and AG1478 on Enterovirus Replication
Authors: van der Linden Lonneke , van der Schaar Hilde M. , Lanke Kjerstin H. W. , Neyts Johan , van Kuppeveld Frank J. M.
Published: May 2010

The genus
Enterovirus
, belonging to the family
Picornaviridae
, includes well-known pathogens, such as poliovirus, coxsackievirus, and rhinovirus. Brefeldin A (BFA) impedes replication of several enteroviruses through inhibition of Golgi-specific BFA resistance factor 1 (GBF1), a regulator of secretory pathway integrity and transport. GBF1 mediates the GTP exchange of Arf1, which in activated form recruits coatomer protein complex I (COP-I) to Golgi vesicles, a process important in transport between the endoplasmic reticulum and Golgi vesicles. Recently, the drugs AG1478 and Golgicide A (GCA) were put forward as new inhibitors of GBF1. In this study, we investigated the effects of these putative GBF1 inhibitors on secretory pathway function and enterovirus replication. We show that both drugs induced fragmentation of the Golgi vesicles and caused dissociation of Arf1 and COP-I from Golgi membranes, yet they differed in their effect on GBF1 localization. The effects of AG1478, but not those of GCA, could be countered by overexpression of Arf1, indicating a difference in their molecular mechanism of action. Consistent with this idea, we observed that GCA drastically reduced replication of coxsackievirus B3 (CVB3) and other human enterovirus species, whereas AG1478 had no effect at all on enterovirus replication. Time-of-addition studies and analysis of RNA replication using a subgenomic replicon both showed that GCA suppresses RNA replication of CVB3, which could be countered by overexpression of GBF1. These results indicate that, in contrast to AG1478, GCA inhibits CVB3 RNA replication by targeting GBF1. AG1478 and GCA may be valuable tools to further dissect enterovirus replication.
The Evolution of Sex: a Perspective from the Fungal Kingdom
Authors: Lee Soo Chan , Ni Min , Li Wenjun , Shertz Cecelia , Heitman Joseph
Published: May 2010

Sex is shrouded in mystery. Not only does it preferentially occur in the dark for both fungi and many animals, but evolutionary biologists continue to debate its benefits given costs in light of its pervasive nature. Experimental studies of the benefits and costs of sexual reproduction with fungi as model systems have begun to provide evidence that the balance between sexual and asexual reproduction shifts in response to selective pressures. Given their unique evolutionary history as opisthokonts, along with metazoans, fungi serve as exceptional models for the evolution of sex and sex-determining regions of the genome (the mating type locus) and for transitions that commonly occur between outcrossing/self-sterile and inbreeding/self-fertile modes of reproduction. We review here the state of the understanding of sex and its evolution in the fungal kingdom and also areas where the field has contributed and will continue to contribute to illuminating general principles and paradigms of sexual reproduction.
Essential Biological Processes of an Emerging Pathogen: DNA Replication, Transcription, and Cell Division inAcinetobacterspp
Authors: Robinson Andrew , Brzoska Anthony J. , Turner Kylie M. , Withers Ryan , Harry Elizabeth J. , Lewis Peter J. , Dixon Nicholas E.
Published: May 2010

Within the last 15 years, members of the bacterial genusAcinetobacterhave risen from relative obscurity to be among the most important sources of hospital-acquired infections. The driving force for this has been the remarkable ability of these organisms to acquire antibiotic resistance determinants, with some strains now showing resistance to every antibiotic in clinical use. There is an urgent need for new antibacterial compounds to combat the threat imposed byAcinetobacterspp. and other intractable bacterial pathogens. The essential processes of chromosomal DNA replication, transcription, and cell division are attractive targets for the rational design of antimicrobial drugs. The goal of this review is to examine the wealth of genome sequence and gene knockout data now available forAcinetobacterspp., highlighting those aspects of essential systems that are most suitable as drug targets.Acinetobacterspp. show several key differences from other pathogenic gammaproteobacteria, particularly in global stress response pathways. The involvement of these pathways in short- and long-term antibiotic survival suggests thatAcinetobacterspp. cope with antibiotic-induced stress differently from other microorganisms.
Clouds and cloud immersion alter photosynthetic light quality in a temperate mountain cloud forest
Authors: Reinhardt Keith , Smith William K. , Carter Gregory A.
Published: May 2010

Light quantity and quality strongly influence plant ecophysiology, especially in cloud forests. Only a few studies have investigated the quantitative effects of clouds on incident sunlight, and almost no studies have explored the specific effects of cloud immersion on light quality. In the present study, the spectral quantity and quality of sunlight during sunny (S), low cloud (LC), and cloud immersed (CI) days in open canopy and understory microsites are compared in high elevation cloud forests in the southern Appalachian Mountains, USA. Cumulative daily radiation measured on CI days was 85% less than on S days at open sites, but about the same as S days in understory sites. LC and CI conditions enhanced blue light by 5%–15% in open sites, but decreased it by 25%–60% in understory sites. In cloudy conditions, red light decreased by 6%–11% in open areas and increased by 25%–30% in understory sites. Cloudiness also resulted in important changes in spectral band ratios, and was associated with light diffusivity. Thus, cloud regimes had a strong influence on the photosynthetic light quality of incident sunlight.
Phyllotaxis of the palmEuterpe oleraceaMart. at the level of the shoot apical meristem
Authors: Barabé Denis , Bourque Laura , Yin Xiaofeng , Lacroix Christian
Published: May 2010

Previous studies on palm phyllotaxis deal mainly with the mature trunk. The goals of this study are (i) to determine the relationship between the number of parastichies, the divergence angle, and the plastochrone ratio at the level of the shoot apical meristem; (ii) to examine whether there are fluctuations in the divergence angle; (iii) to interpret the significance of phyllotactic parameters with respect to the mode of growth of the apex. The tubular base of the leaf primordium is more or less asymmetrical, and completely surrounds the shoot apical meristem. The phyllotactic system corresponds to a (2, 3) conspicuous parastichy pair. The mean divergence angle per apex varies between 126.9° ± 9.3° (mean ± SD) and 135. 8° ± 8.0°. Divergence angles for all apices fluctuate within a range of 115.89° to 157.33°. The mean plastochrone ratios between apices varies from 1.35 ± 0.18 to 1.58 ± 0.12. The plastochrone ratio at each plastochrone for all apices ranges from 1.09 to 2.00. There is no correlation between the angle of divergence and the plastochrone ratio. There is a fluctuation in the value of the divergence angle that falls within the range predicted by the fundamental theorem of phyllotaxis. The high value of the ratio of the diameter of leaf primordia over the diameter of the apex, and the long plastochrone might explain the lack of correlation between certain phyllotactic parameters.
γ-Aminobutyrate transaminase limits the catabolism of γ-aminobutyrate in cold-stressed Arabidopsis plants: insights from an overexpression mutant
Authors: Simpson Jeffrey P. , Clark Shawn M. , Portt Andrea , Allan Wendy L. , Makhmoudova Amina , Rochon Amanda , Shelp Barry J.
Published: May 2010

We tested the hypothesis that γ-aminobutyrate transaminase (GABA-T) regulates the supply of succinate semialdehyde for succinate semialdehyde dehydrogenase or NADPH-dependent glyoxylate/succinate semialdehyde reductase 1 (GLYR1) during stress. Constitutive overexpression (OX) lines of GABA-T were generated in Arabidopsis via the floral-dip method for Agrobacterium-mediated transformation. Polymerase chain reaction enabled selection of four transgenic lines with higher GABA-T transcript levels than the wild-type (WT), but assay of cell-free leaf extracts revealed that only OX1 had elevated GABA-T activity. Brief cold treatments (4 °C exposure for 20 min or 1 h in the dark) increased leaf GABA concentrations in both the WT and OX1, but the concentrations in OX1 were consistently lower. These findings confirm that GABA-T limits the catabolism of GABA when its production is stimulated by stress, and suggest a bioengineering strategy for improving the availability of succinate semialdehyde for the Krebs cycle or GLYR1, a potential redox-modulating reaction during stress.
Mycorrhizal and dark septate fungal associations in gingers and spiral gingers
Authors: Uma Eswaranpillai , Muthukumar Thangavelu , Sathiyadash Kullaiyan , Muniappan Vellaisamy
Published: May 2010

Associations of arbuscular mycorrhizal (AM) and dark septate endophyte (DSE) fungi with 38 ginger species (Zingiberaceae) and two spiral gingers (Costaceae) were investigated. All species were mycorrhizal, and the status of 35 gingers and one spiral ginger has been investigated for the first time. The extent of AM colonization and the root length colonized by AM structures varied significantly among species. AM structures were also observed in non-root portions such as scale leaves. Arbuscular mycorrhizae were of either Arum-type or intermediate- type; AM-type is reported for the first time in 37 species. Spores of 16 morphotypes belonging to 4 genera ( Glomus , Acaulospora , Scutellospora , Racocetra ) were identified from the rhizosphere soil. DSE colonization was found in 22 gingers and 2 spiral gingers. Twenty-two plant species are reported as hosts of DSE fungi for the first time. The extent of total AM and DSE colonization was not correlated.
Ectomycorrhizal community composition of Pinus tabulaeformis assessed by ITS-RFLP and ITS sequences
Authors: Wang Qin , Guo Liang-Dong
Published: May 2010

Ectomycorrhizal (ECM) fungal composition was examined in a Pinus tabulaeformis Carr. forest. A total of 28 root samples of P. tabulaeformis were collected in June and September. Thirty-five ECM morphotypes were identified according to ECM morphological characters, and 26 ECM fungi were identified based on the analyses of ITS-RFLP and ITS sequences. Tomentella , Sebacina , and Tuber were common genera, and Atheliaceae sp., Lactarius deliciosus , Tomentella ferruginea , and Tomentella sp. 3 were dominant species. Of these ECM fungi, 13 were found in June, 19 in September, and 6 during both sampling times. Atheliaceae sp. and T. ferruginea were the dominant fungi both in June and September. Lactarius deliciosus was dominant in June, but rare in September. Tomentella sp. 3 was dominant in September but rare in June.