bluuebunny/crossref_metadata_2025 huggingface dataset:
Changes in Sexual Function after Treatment for Prolapse Are Related to the Improvement in Body Image Perception
Authors: Lowenstein Lior , Gamble Tondalaya , Sanses Tatiana V. Deniseiko , Van Raalte Heather , Carberry Cassie , Jakus Sharon , Pham Thythy , Nguyen Aimee , Hoskey Kay , Kenton Kimberly
Published: November 2009

Introduction

In a previous study, sexual function was related to a woman’s self-perceived body image and degree of bother from pelvic organ prolapse (POP).

Aims

To evaluate sexual function, prolapse symptoms, and self-perceived body image 6 months following treatment for POP and to explore differences in body image perception and sexual function following conservative and surgical treatment for POP.

Methods

After institutional review board approval, consecutive women with ≥stage II POP were invited to participate. In addition to routine urogynecologic history and physical examination, including Pelvic Organ Prolapse Quantification (POP-Q), the participants completed three validated questionnaires before, and 6 months after, treatment for POP: Pelvic Organ Prolapse/Urinary Incontinence Sexual Questionnaire, Modified Body Image Perception Scale and Prolapse subscale of Pelvic Floor Distress Inventory to assess condition specific bother from POP.

Main Outcome Measures

Changes in sexual function and body image perception following treatment for POP.

Results

A total of 235 women with a mean age of 62 ± 12 years returned for a 6-month follow-up. The majority of our participants had surgical repair for POP (88%). At 6-month follow-up visits, the patients reported significant improvement in sexual function from baseline (33 ± 0.6 vs. 43 ± 0.8, respectively P < 0.0001). Improvement in sexual function, as measured by PISQ-12, was not significant among sexually active patients treated with a pessary compared with those treated surgically (–2.5 ± 5.5 vs. 11.5 ± 1, respectively P < 0.0001). A multivariate linear regression model demonstrated that body mass index and changes in body image perception were the only independent factors associated with changes in PISQ score following POP treatment (β = −0.5, P < 0.01 and β = −0.4, P < 0.03, respectively).

Conclusions

Resolution of POP symptoms after treatment improves women’s self-perceived body image and sexual function. Not surprisingly, pessary is less effective in improving sexual function compared with surgical repair of POP.
A giant planet in orbit around a magnetic-braking hibernating cataclysmic variable
Authors: Qian S.-B. , Liao W.-P. , Zhu L.-Y. , Dai Z.-B. , Liu L. , He J.-J. , Zhao E.-G. , Li L.-J.
Published: November 2009

Detections of Jupiter-like giant planets in orbit around short-period white dwarf binaries should provide insight into the formation and evolution of circumbinary planets (planets orbiting both components of short-period binaries), as well as into the ultimate fate of planets and the late evolutionary stage of binary stars (e.g. the evolution of the common envelope). However, to date no planets have been detected as companions to such close binaries. Here, we report the discovery of a giant planet orbiting the only known hibernating cataclysmic variable (CV), QS Vir, with a period of 7.86 yr. We analysed the variations of the orbital period of the eclipsing white dwarf–red dwarf binary, and a very small-amplitude cyclic change is found to be superimposed on a long-term period decrease. The period oscillation has the smallest amplitude among close binary stars and can be plausibly interpreted as the light-travel time effect via the presence of a third body. We found that the tertiary component is a giant planet with a mass of ∼6.4 MJupiter at a distance of ∼4.2 astronomical units (au) from the binary. The continuous decrease is explained as angular momentum loss via magnetic braking which is driving the evolution of the hibernating CV into a normal cataclysmic binary.
Escherichia coli: Dominance of Red Light over Other Visible Light Sources in Establishing Viable but Nonculturable State
Authors: İdil Önder , Özkanca Reşit , Darcan Cihan , Flint Ken P.
Published: November 2009

In this study, the effect of UV‐A and different wavelengths of visible light irradiations combined with or without a photosensitizer (methylene blue, MB) on the establishment of viable but nonculturable (VBNC) state in Escherichia coli was investigated. Survival of the E. coli was investigated by measuring plate counts, respiring cell count (RCC), direct viable count (DVC) and total counts over a period of up to 72 h. The inhibition rates of various light sources in the presence or absence of MB on E. coli in seawater were ranked in the order UV‐A>red light>white light>blue light>green light (from greatest to least activation). E. coli survived for 10.2, 19.0, 21.3 and 24.04 h under exposure to red, white, blue and green light and for 6.8 h under exposure to UV‐A in the presence of MB according to t99. Although the VC declined to undetectable levels in a relatively short time, the RCC showed that some cells were still capable of respiration and, therefore, are assumed to have entered the VBNC phase. This is the first time that red light has been shown to have a stronger effect on E. coli survival and VBNC than white, green and blue light in seawater environment.
New Discoveries From the Bilingual Brain and Mind Across the Life Span: Implications for Education
Authors: Petitto Laura‐Ann
Published: November 2009

We discuss the fruits of educational neuroscience research from our laboratory and show how the typical maturational timing milestones in bilingual language acquisition provide educators with a tool for differentiating a bilingual child experiencing language and reading delay versus deviance. Further, early schooling in two languages simultaneously affords young bilingual children a reading advantage and may also ameliorate the negative effect of low socioeconomic status on literacy. Using powerful brain imaging technology, functional Near Infrared Spectroscopy, we provide a first‐time look into the developing brains of bilingual as comapred to monolingual children. We show unequivocally that the age of first bilingual exposure is a vital predictor of bilingual language and reading mastery. Accounts that promote later dual language and reading instruction, or those that assert human brain development is unrelated to bilingual language mastery, are not supported by the present findings. We discuss the implications for education, teachers, and developmental brain sciences.
A 30‐Minute Physical Education Program Improves Students' Executive Attention
Authors: Kubesch Sabine , Walk Laura , Spitzer Manfred , Kammer Thomas , Lainburg Alyona , Heim Rüdiger , Hille Katrin
Published: November 2009

Physical activity is not only beneficial to physical health but also to cognitive functions. In particular, executive functions that are closely related to learning achievement can be improved by acute and recurring physical activity. We examined the effects of a single 30‐min physical education program in contrast to a 5‐min movement break on working memory, cognitive flexibility, and inhibition of attention and behavioral tendencies of eighty‐one 13‐ to 14‐year‐old students in grade 7 in Germany. Results indicate that the maintenance of on‐task attention in the face of distraction was improved by an aerobic endurance exercise‐based physical education program but not by a short aerobic movement break. This suggests that the duration of a school sports program is decisive for improving students' executive attention.
Perineurioma of the mandibular dental nerve: a case report and review of the literature
Authors: Vencio E.F. , Cheim Jr A.P. , Alencar R.C. , Alencar W.M. , Dias Filho A.A.
Published: November 2009

Perineurioma is an uncommon benign tumour of the nerve sheath composed exclusively of well‐differentiated perineurial cells. It usually affects the limbs or trunk; intraoral lesions are uncommon. A rare case of perineurioma is described involving the mandibular dental nerve in a 59‐year‐old female, microscopically characterised by a combined pattern of intraneural and soft tissue perineurioma. The patient was referred for treatment of a lesion in the mandible. Panoramic radiography revealed a well‐delimited osteolytic lesion with radiopaque halo in an edentulous region of the left mandible. Occlusal view showed no expansion of cortical bone. A surgical procedure for incisional biopsy disclosed a connection between the tumour and the alveolar inferior nerve. Grossly, an enlargement of the affected nerve was described. Microscopic examination showed tumour cells arranged in pseudo‐onion bulb structures, with scattered collagen bundles, and areas with a whorled (storiform) arrangement. Many concentric whorls of spindle cells exhibiting thin, elongated eosinophic processes were seen. Immunohistochemical study exhibited strong positivity for epithelial membrane antigen. We report a rare case of perineurioma affecting the mandibular dental nerve showing both intraneural and soft tissue microscopic patterns and discuss morphological aspects for differential diagnosis.
Peripheral keratocystic odontogenic tumour: a case report
Authors: Jinbu Y. , Ueno Y. , Obi Y. , Ikeda K. , Kusama M. , Tsukinoki K.
Published: November 2009

Peripheral keratocystic odontogenic tumour is a rare disease occurring in the gingiva and displaying histopathological characteristics similar to keratocystic odontogenic tumour. The patient was a 63‐year‐old man complaining of gingival swelling in the mandibular premolar region. A raised mass lesion 15 mm in diameter covered with normal epithelium was observed on the buccal gingiva of the left mandibular premolar region. Panoramic radiography and computed tomography excluded intraosseous cyst or tumour. Needle aspiration examination revealed paste‐like, pale yellow contents. The raised lesion was surgically resected. Histopathologically, the cyst wall comprised squamous epithelium with parakeratosis. Peripheral keratocystic odontogenic tumour was diagnosed.
p53 mutations in keratocystic odontogenic tumour
Authors: Kuroyanagi N. , Machida J. , Sakuma H. , Miyabe S. , Hashimoto O. , Yokoi M. , Warnakulasuriya S. , Nagao T. , Shimozato K.
Published: November 2009

Aim:  The abrogation of the function for the tumour suppressor gene p53 has been suggested as one of the most common genetic alterations in neoplasia. We evaluated p53 mutations, clinico‐pathological and immunohistochemical findings in patients with sporadic keratocystic odontogenic tumours (KCOTs).

Material and methods:  A total of 32 patients with KCOT treated surgically were enrolled in this study. We performed mutation analysis of p53 using archival formalin‐fixed paraffin‐embedded tissue. The immunohistological technique was employed to identify TP53 expression. P53 mutations in exons 5–8 were evaluated by direct sequencing. These results were compared with the characteristics of each tumour.

Results:  Forty‐four percent of the cysts showed overexpression of TP53. P53 mutations were detected in 10 cases (31.3%) of 32 KCOTs; two cases within exon 5, two cases within exon 8, five cases within exon 7 and one case in both exons 5 and 7. Five missense mutations (R248G, D247K, Q136R, V272M, V143A) and four silence mutations (138A, 158R, 247D, 279G,) were detected. P53 mutation was detected in two (50%) out of four recurrence cases and eight (28.5%) out of 28 non‐recurrence cases (P = 0.572). R248G mutation was the most common in KCOTs (15.6%). We did not detect any remarkable associations of these cases carrying p53 mutations to clinico‐pathological and immunohistochemical characteristics.

Conclusion:  The results implied that the p53 gene alterations are involved in the development of KCOT.
Valuing and Caring
Authors: SEIDMAN JEFFREY
Published: November 2009

What is it to “value” something, in the semi‐technical sense of the term that Gary Watson establishes? I argue that valuing something consists incaringabout it. Caring involves not only emotional dispositions of the sort that Agnieszka Jaworska has elaborated, but also a distinctivecognitivedisposition – namely, a (defeasible) disposition to believe the object cared about to be a source of agent‐relative reasons for action and for emotion. Understood in this way, an agent's carings have a stronger claim to “speak for” her as her values than do other attitudes that have been proposed for this role. In particular, an agent's carings establish more robust psychological continuities and cross‐temporal connections than do self‐governing policies of the sort that Michael Bratman has described; and they forge diachronic coherence not just in her deliberation and action, as self‐governing policies do, but also in her cognitive and emotional life. An agent's carings thus help to constitute her identity as a temporally persistingsubject. Self‐governing policies are at bestersatz‐values, which an agent may choose to adopt when she finds that her proper values – her cares – leave her course underdetermined.
Pelvic Autonomic Dysfunction without Tetraparesis: A Sequel of Rubella‐related Acute Longitudinal Myelitis
Authors: TAKAHASHI Osamu , SAKAKIBARA Ryuji , KISHI Masahiko , OGAWA Emina , TSUNOYAMA Kuniko , SUGIYAMA Megumi , TOMARU Takanobu , UCHIYAMA Tomoyuki , YAMANISHI Tomonori
Published: November 2009

Objectives: Pelvic autonomic dysfunction without tetraparesis can be a sequel of acute longitudinal myelitis.

Methods: A 25‐year‐old woman had suffered from pelvic dysfunction (urinary retention and severe constipation) without tetraparesis for 5 months as a sequel of rubella‐related acute longitudinal myelitis at the spinal segments of C2 to T9. She still had brisk deep tendon reflexes and extensor plantar responses. We performed autonomic function tests in the patient.

Results: Pelvic autonomic function tests revealed acontractile detrusor and unrelaxing sphincter, suggesting the shock phase of a suprasacral spinal cord lesion; as well as loss of voluntary sphincter contraction, prolonged colonic transit time, decreased spontaneous phasic contractions, and paradoxical sphincter contraction on defecation, suggesting a suprasacral cord lesion.

Conclusion: These findings indicated that the lateral medullary reticulospinal tracts just inside the pyramidal tract, for example, the descending pathways to the bladder and bowel, and the adjacent corticospinal tract to the sphincter, were selectively affected in our case.
Molecular Detection of Leishmania infantum in Wild Rodents (Rattus norvegicus) in Greece
Authors: Papadogiannakis E. , Spanakos G. , Kontos V. , Menounos P. G. , Tegos N. , Vakalis N.
Published: November 2009

Leishmania infantum and Leishmania tropica are the species responsible for visceral leishmaniasis and cutaneous leishmaniasis respectively. In Greece, both diseases are endemic. The dog is considered the main reservoir of L. infantum, whereas the role of other animals for both L. infantun and L. tropica is unknown. Spleens from wild Rattus norvegicus, live trapped in Greece, were examined for the presence of Leishmania parasites by PCR. Out of 16 samples examined, only one was found positive for L. infantum with scant amount of parasitic DNA present. This is the first documented case of detection of L. infantum in R. norvegicus in Greece. The results of this preliminary study indicate that R. norvegicus is unlikely to be a reservoir for Leishmania parasites in Greece.
The Effect of Intermetallic Growth on Bump Pull Test Responses of SAC105 Solder Bumps
Authors: Amistoso Jose Omar S. , Amorsolo Alberto V.
Published: November 2009

Cold bump pull tests performed on wafer level chip scale packages using SAC105 solder bumps show an increase in the occurrence of brittle failure modes with aging temperature and time. Fast intermetallic growth at 0–1000 h can be attributed to (Cu,Ni)6Sn5, while the decrease in intermetallic growth rate at t>1000 h can be attributed to diffusion processes leading to (Cu,Ni)6Sn5 and (Ni,Cu)3Sn4 formation and growth. Ni diffuses toward the solder bulk and saturates at 175–200°C, while Cu diffuses from the under bump metallization (UBM) toward the solder bump at 125–150°C. Interactions between Cu and Ni atoms lead to saturation of their atomic % gradients due to intermetallic formation. Sn diffusion from the solder toward the UBM occurs at 125–150°C. The activation energy for total intermetallic growth was calculated at 0.2 eV.
Improved Solder Joint Fatigue Models Through Reduced Geometry Dependence of Empirical Fits
Authors: Bhate D. , Subbarayan G. , Zhao J. , Gupta V. , Edwards D.
Published: November 2009

Predicting the fatigue life of solder interconnections is a challenge due to the complex nonlinear behavior of solder alloys, the importance of the load history, and the wide variation in package constructions employed by the microelectronics industry. While material behavior can be captured by means of valid constitutive models, the empirical failure models in common use are strongly dependent on package construction due to the fact that they are derived from tests of solder joints with heterogeneous stress and microstructural variation. Despite this limitation, the empirical models have been used to identify reliable choices among package design alternatives. In this technical brief, some of the key limitations that the user of empirical models must be aware of are addressed. Physical arguments are proposed in order to justify the appropriate assumptions that may be made in the life model. Using data from two different sources in the literature, the constants in the empirical models are shown to be dependent on the solder joint geometry. Alternative fits to experimental data that reduce the dependence of the fits on solder joint geometry are proposed.
Numerical Simulation of Thermal Conductivity of Particle Filled Epoxy Composites
Authors: Zeng Jun , Fu Renli , Agathopoulos Simeon , Zhang Shaodong , Song Xiufeng , He Hong
Published: November 2009

A finite element method was developed to predict the effective thermal conductivity of particle filled epoxy composites. Three-dimensional models, which considered the effect of filler geometry, filler aspect ratio, conductivity ratio of filler to matrix, and interfacial layer were used to simulate the microstructure of epoxy composites for various filler volume fractions up to 30%. The calculated thermal conductivities were compared with results from existing theoretical models and experiments. Numerical estimation of ellipsoids-in-cube model accurately predicted thermal conductivity of epoxy composites with alumina filler particles. The number of length division during mesh process and particle numbers used in the finite element analysis affect the accuracy of calculated results. At a given value of filler content, the numerical results indicated a ratio of conductivity of filler to matrix for achieving the maximum thermal conductivity.
Finite Element Modeling of Simultaneous Ultrasonic Bumping With Au Balls
Authors: Song Wan Ho , Karimi Ali , Huang Yan , Mayer Michael , Zhou Norman , Jung Jae Pil
Published: November 2009

Bumping of microcircuits and substrates establishes interconnect points required for subsequent bonding of microelectronic components, allowing for power and data distribution. Simultaneous ultrasonic bonding of individual Au balls promises to accelerate bumping processes and is studied using a finite element model. The model covers the static forces at the end of a successful bonding operation and analyzes the interfacial stresses between bumps and substrate. The modeling shows the vertical forces acting on the bumps when a lateral displacement of the bonding tool is applied. When designing a practical bonding application, the control of such vertical forces is recommended. A sensitivity analysis is conducted to study the effect of the main factors on the model responses. This analysis reveals that variations in bump height and bonding tool elastic modulus are the major factors affecting the forces on the bumps.
Observation of Half-Quantum Vortices in an Exciton-Polariton Condensate
Authors: Lagoudakis K. G. , Ostatnický T. , Kavokin A. V. , Rubo Y. G. , André R. , Deveaud-Plédran B.
Published: November 2009

Quantum Division

The notion of quantum mechanics is that variables are expressed as integer values. In quantum fluids, for instance, vortices are quantized in terms of the polarization and phase shifts observed as multiples of 2π, that is, full rotations of each variable around the vortex core. Theoretical work has predicted that in some instances there should be half-quantum vortices, where circumventing the vortex core is characterized by just π rotations of the phase and polarization. By observing exciton-polariton condensates

Lagoudakis
et al.

(p.
974
) present evidence for the existence of these long-predicted half-quantum vortices.
Widespread Occurrence of Self-Cleaving Ribozymes
Authors: Webb Chiu-Ho T. , Riccitelli Nathan J. , Ruminski Dana J. , Lupták Andrej
Published: November 2009

Once thought to be an oddity, small catalytic RNAs have been found in a wide range of organisms.
Partitioning Recent Greenland Mass Loss
Authors: van den Broeke Michiel , Bamber Jonathan , Ettema Janneke , Rignot Eric , Schrama Ernst , van de Berg Willem Jan , van Meijgaard Erik , Velicogna Isabella , Wouters Bert
Published: November 2009

GRACE and Movement Together

Recent measurements of the rate of mass loss from the Greenland ice sheet vary approximately by a factor of three. Resolving these discrepancies is essential for determining the current mass balance of the ice sheet and to project sea level rise in the future.

Van den Broeke
et al.

(p.
984
) obtained consistent estimates from two independent methods, one based on observations of ice movement combined with model calculations and the other on remote gravity measurements made by the GRACE (Gravity Recovery and Climate Experiment) satellites. The combination of these approaches also resolves the separate contributions of surface processes and of ice dynamics, the two major routes of ice mass loss.
Ruth L. Kirschstein (1926–2009)
Authors: Schachman Howard K. , Cassman Marvin
Published: November 2009

The leadership, political skills, and commitment of the first woman director at the U.S. National Institutes of Health are unrivaled.
On Becoming a Scientist
Authors: Alberts Bruce
Published: November 2009

One normally becomes a scientist through a series of apprenticeships, pursuing research in laboratories directed by established scientists. My own scientific mentors were Jacques Fresco and Paul Doty at Harvard, where I learned not only technical skills but also how to think and function as a scientist. Both from them, and by making my own mistakes,
*
I learned how to identify important problems, how to think critically, and how to design effective research strategies. Because so much of one's scientific future is shaped by early experiences, it is critical that beginning scientists select their mentors wisely. Unfortunately, what constitutes a “good” choice is not always obvious. Here I offer some personal advice to help young scientists make these tough decisions wisely.
Wellcome Trust to Shift From Projects to People
Authors: Kaiser Jocelyn
Published: November 2009

This week, Wellcome Trust officials announced that they are phasing out their biomedical research grants for U.K. scientists, which consists of 3-year to 5-year awards focused on a specific problem, and instead will put that money into a new program called Investigator Awards. The awards will be bigger, more flexible, and longer—up to 7 years.
No Sign Yet of Himalayan Meltdown, Indian Report Finds
Authors: Bagla Pallava
Published: November 2009

Himalayan glaciers are beating a rapid retreat in the face of global warming, according to a flurry of recent reports by BBC and other mass media. But the picture is more complex—and poses scientific puzzles, according to a review of satellite and ground measurements released by India's Ministry of Environment and Forests earlier this week.
Could Glacier Research Help Thaw Himalayan Standoff?
Authors: Bagla Pallava
Published: November 2009

Because of lingering tensions over a swath of the Himalayas that is claimed by both China and India, a diplomatic initiative to get Chinese and Indian scientists working together on glaciers has quietly been put on ice.
ITER Blueprints Near Completion, But Financial Hurdles Lie Ahead
Authors: Clery Daniel
Published: November 2009

As staff put finishing touches to the fusion project's final design, member governments mull over the latest cost estimates and prepare to raid piggybanks.
Comparison of Three Antigenic Extracts of
Eurotium amstelodami
in Serological Diagnosis of Farmer's Lung Disease
Authors: Roussel Sandrine , Reboux Gabriel , Rognon Bénédicte , Monod Michel , Grenouillet Frédéric , Quadroni Manfredo , Fellrath Jean-Marc , Aubert John-David , Dalphin Jean-Charles , Millon Laurence
Published: November 2009

In France and Finland, farmer's lung disease (FLD), a hypersensitivity pneumonitis common in agricultural areas, is mainly caused by
Eurotium
species. The presence of antibodies in patients' serum is an important criterion for diagnosis. Our study aimed to improve the serological diagnosis of FLD by using common fungal particles that pollute the farm environment as antigens. Fungal particles of the
Eurotium
species were observed in handled hay. A strain of
Eurotium amstelodami
was grown
in vitro
using selected culture media; and antigen extracts from sexual (ascospores), asexual (conidia), and vegetative (hyphae) forms were made. Antigens were tested by enzyme-linked immunosorbent assay (ELISA), which was used to test for immunoglobulin G antibodies from the sera of 17 FLD patients, 40 healthy exposed farmers, and 20 nonexposed controls. The antigens were compared by receiver operating characteristic analysis, and a threshold was then established. The ascospores contained in asci enclosed within cleistothecia were present in 38% of the hay blades observed; conidial heads of aspergillus were less prevalent. The same protocol was followed to make the three antigen extracts. A comparison of the results for FLD patients and exposed controls showed the area under the curve to be 0.850 for the ascospore antigen, 0.731 for the conidia, and 0.690 for the hyphae. The cutoffs that we determined, with the standard deviation for measures being taken into account, showed 67% for sensitivity and 92% for specificity with the ascospore antigen. In conclusion, the serological diagnosis of FLD by ELISA was improved by the adjunction of ascospore antigen.
Highly Penicillin-Resistant Multidrug-Resistant Pneumococcus-Like Strains Colonizing Children in Oeiras, Portugal: Genomic Characteristics and Implications for Surveillance
Authors: Simões Alexandra S. , Sá-Leão Raquel , Eleveld Marc J. , Tavares Débora A. , Carriço João A. , Bootsma Hester J. , Hermans Peter W. M.
Published: November 2009

While performing surveillance studies in Oeiras, Portugal, designed to describe the impact of pneumococcal conjugate vaccine on colonization, we observed an increase from 0.7% in 2003 to 5% in 2006 in the prevalence of penicillin resistance (MIC of 2 to 6 mg/liter) among presumptively identified pneumococcal isolates. Although 15 of the 22 penicillin-resistant isolates obtained in 2006 were optochin resistant, they were bile soluble and thus considered to be bona fide pneumococci. This study aimed to clarify the nature of these isolates by using a combination of phenotypic and genotypic approaches that included routine strategies for pneumococcal identification, multilocus sequence analysis (MLSA), and comparative genomic hybridization (CGH). By MLSA, all isolates were classified as “streptococci of the mitis group” that, however, were distinct from typical
Streptococcus pneumoniae
or
Streptococcus mitis
. A single isolate was identified as
Streptococcus pseudopneumoniae
. CGH confirmed these findings and further indicated that a considerable part of the proposed pneumococcal core genome is conserved in these isolates, including several pneumococcal virulence genes (e.g.,
pavA
,
spxB
,
cbpE
, and
cbpD
). These results suggest that among pneumococci and closely related streptococci, universal unique phenotypic and genetic properties that could aid species identification are virtually impossible to define. In pneumococcal colonization studies, when atypical strains are found, MLSA and CGH are informative tools that can be used to complement routine tests. In our study, after correct identification of the penicillin-resistant true pneumococci, we found that penicillin resistance levels among pneumococci remained stable from 2003 to 2006.
Determining Cystic Fibrosis-Affected Lung Microbiology: Comparison of Spontaneous and Serially Induced Sputum Samples by Use of Terminal Restriction Fragment Length Polymorphism Profiling
Authors: Rogers Geraint B. , Skelton Stuart , Serisier David J. , van der Gast Christopher J. , Bruce Kenneth D.
Published: November 2009

Sampling of the lower airways of the adult cystic fibrosis (CF) lung has received insufficient detailed consideration, with the importance of sampling strategies for bacteriological outcome not known. Although spontaneously expectorated sputum (SES) samples are often used for diagnostic bacteriological analysis, induced sputum (IS) methods have advantages. This study examined whether significant differences in bacterial content were detected when using a culture-independent, molecular profiling technique to analyze SES or IS samples. Moreover, this work examined what trends relating to bacterial species distributions and reproducibility were found in sequentially induced sputum samples and what implications this has for pathogen detection. Terminal restriction fragment length polymorphism (T-RFLP) analysis was performed on a SES sample and 4 subsequent IS samples taken at 5-min intervals from 10 clinically stable, adult CF patients. This was repeated over 3 sampling days, with variability between samples, induction periods, and sampling days determined. A diverse range of bacterial species, including potentially novel pathogens, was found. No significant difference in bacterial content was observed for either SES or serial IS samples. On average, the analysis of a single sample from any time point resolved ∼58% of total bacterial diversity achieved by analysis of an SES sample and 4 subsequent IS samples. The reliance on analysis of a single respiratory sample was not sufficient for the detection of recognized CF pathogens in all instances. Close correlation between T-RFLP and microbiological data in the detection of key species indicates the importance of these findings in routine diagnostics for the detection of recognized and novel CF pathogens.
Detection of Occult
Scedosporium
Species in Respiratory Tract Specimens from Patients with Cystic Fibrosis by Use of Selective Media
Authors: Blyth C. C. , Harun A. , Middleton P. G. , Sleiman S. , Lee O. , Sorrell T. C. , Meyer W. , Chen S. C. A.
Published: November 2009

Respiratory samples from cystic fibrosis outpatients were cultured on Sabouraud's dextrose agar (SABD) containing antibiotics, Mycosel, and
Scedosporium
-selective medium (SceSel+). Thirty-two (14.7%) of 218 specimens from 11/69 (15.9%) patients yielded a
Scedosporium
sp., most frequently
Scedosporium aurantiacum
(17/218).
Scedosporium
was recovered on SceSel+, Mycosel, and SABD from 90.6%, 50.0%, and 46.9% of the specimens tested, respectively.
Commercial DNA Probes for Mycobacteria Incorrectly Identify a Number of Less Frequently Encountered Species
Authors: Tortoli Enrico , Pecorari Monica , Fabio Giuliana , Messinò Massimino , Fabio Anna
Published: November 2009

Although commercially available DNA probes for identification of mycobacteria have been investigated with large numbers of strains, nothing is known about the ability of these probes to identify less frequently encountered species. We analyzed, with INNO LiPA MYCOBACTERIA (Innogenetics) and with GenoType Mycobacterium (Hein), 317 strains, belonging to 136 species, 61 of which had never been assayed before. INNO LiPA misidentified 20 taxa, the majority of which cross-reacted with the probes specific forMycobacterium fortuitumand theMycobacterium avium-Mycobacterium intracellulare-Mycobacterium scrofulaceumgroup. GenoType misidentified 28 taxa, most of which cross-reacted withM. intracellulareandM. fortuitumprobes; furthermore, eight species were not recognized as members of the genusMycobacterium. Among 54 strains investigated with AccuProbe (Gen-Probe), cross-reactions were detected for nine species, with the probes aiming at theM. aviumcomplex being most involved in cross-reactions.
Internal Transcribed Spacer Region Sequence Heterogeneity in
Rhizopus microsporus
: Implications for Molecular Diagnosis in Clinical Microbiology Laboratories
Authors: Woo Patrick C. Y. , Leung Shui-Yee , To Kelvin K. W. , Chan Jasper F. W. , Ngan Antonio H. Y. , Cheng Vincent C. C. , Lau Susanna K. P. , Yuen Kwok-Yung
Published: November 2009

Although internal transcribed spacer region (ITS) sequence heterogeneity has been reported in a few fungal species, it has very rarely been reported in pathogenic fungi and has never been described in
Mucorales
, causes of the highly fatal mucormycosis. In a recent outbreak investigation of intestinal mucormycosis due to
Rhizopus microsporus
infection in patients with hematological malignancies, PCR of the ITS of four of the 28
R. microsporus
strains, P11, P12, D3-1, and D4-1, showed thick bands at about 700 bp. Direct sequencing of the purified bands showed frequent double peaks along all of the sequence traces and occasional triple peaks for P12, D3-1, and D4-1. The thick bands of the four
R. microsporus
strains were purified and cloned. Sequencing of 10 clones for each strain revealed two different ITS sequences for P11 and three different ITS sequences for P12, D3-1, and D4-1. Variations in ITS sequence among the different ribosomal DNA (rDNA) operons in the same strain were observed in only ITS1 and ITS2 and not the 5.8S rDNA region. One copy of P11, P12, and D4-1, respectively, and one copy of P11, P12, D3-1, and D4-1, respectively, showed identical sequences. This represents the first evidence of ITS sequence heterogeneity in
Mucorales
. ITS sequence heterogeneity is an obstacle to molecular identification and genotyping of fungi in clinical microbiology laboratories. When thick bands and double peaks are observed during PCR sequencing of a gene target, such a strain should be sent to reference laboratories proficient in molecular technologies for further identification and/or genotyping.
Antiangiogenic Arming of an Oncolytic Vaccinia Virus Enhances Antitumor Efficacy in Renal Cell Cancer Models
Authors: Guse Kilian , Sloniecka Marta , Diaconu Iulia , Ottolino-Perry Kathryn , Tang Nan , Ng Calvin , Le Boeuf Fabrice , Bell John C. , McCart J. Andrea , Ristimäki Ari , Pesonen Sari , Cerullo Vincenzo , Hemminki Akseli
Published: November 2009

Oncolytic vaccinia viruses have shown compelling results in preclinical cancer models and promising preliminary safety and antitumor activity in early clinical trials. However, to facilitate systemic application it would be useful to improve tumor targeting and antitumor efficacy further. Here we report the generation of vvdd-VEGFR-1-Ig, a targeted and armed oncolytic vaccinia virus. Tumor targeting was achieved by deletion of genes for thymidine kinase and vaccinia virus growth factor, which are necessary for replication in normal but not in cancer cells. Given the high vascularization typical of kidney cancers, we armed the virus with the soluble vascular endothelial growth factor (VEGF) receptor 1 protein for an antiangiogenic effect. Systemic application of high doses of vvdd-VEGFR-1-Ig resulted in cytokine induction in an immunocompromised mouse model. Upon histopathological analysis, splenic extramedullary hematopoiesis was seen in all virus-injected mice and was more pronounced in the vvdd-VEGFR-1-Ig group. Analysis of the innate immune response after intravenous virus injection revealed high transient and dose-dependent cytokine elevations. When medium and low doses were used for intratumoral or intravenous injection, vvdd-VEGFR-1-Ig exhibited a stronger antitumor effect than the unarmed control. Furthermore, expression of VEGFR-1-Ig was confirmed, and a concurrent antiangiogenic effect was seen. In an immunocompetent model, systemic vvdd-VEGFR-1-Ig exhibited superior antitumor efficacy compared to the unarmed control virus. In conclusion, the targeted and armed vvdd-VEGFR-1-Ig has promising anticancer activity in renal cell cancer models. Extramedullary hematopoiesis may be a sensitive indicator of vaccinia virus effects in mice.
Cellular Immune Responses to Severe Acute Respiratory Syndrome Coronavirus (SARS-CoV) Infection in Senescent BALB/c Mice: CD4+T Cells Are Important in Control of SARS-CoV Infection
Authors: Chen Jun , Lau Yuk Fai , Lamirande Elaine W. , Paddock Christopher D. , Bartlett Jeanine H. , Zaki Sherif R. , Subbarao Kanta
Published: November 2009

We characterized the cellular immune response to severe acute respiratory syndrome coronavirus (SARS-CoV) infection in 12- to 14-month-old BALB/c mice, a model that mimics features of the human disease. Following intranasal administration, the virus replicated in the lungs, with peak titers on day 2 postinfection. Enhanced production of cytokines (tumor necrosis factor alpha [TNF-α] and interleukin-6 [IL-6]) and chemokines (CXCL10, CCL2, CCL3, and CCL5) correlated with migration of NK cells, macrophages, and plasmacytoid dendritic cells (pDC) into the lungs. By day 7, histopathologic evidence of pneumonitis was seen in the lungs when viral clearance occurred. At this time, a second wave of enhanced production of cytokines (TNF-α, IL-6, gamma interferon [IFN-γ], IL-2, and IL-5), chemokines (CXCL9, CXCL10, CCL2, CCL3, and CCL5), and receptors (CXCR3, CCR2, and CCR5), was detected in the lungs, associated with an influx of T lymphocytes. Depletion of CD8+T cells at the time of infection did not affect viral replication or clearance. However, depletion of CD4+T cells resulted in an enhanced immune-mediated interstitial pneumonitis and delayed clearance of SARS-CoV from the lungs, which was associated with reduced neutralizing antibody and cytokine production and reduced pulmonary recruitment of lymphocytes. Innate defense mechanisms are able to control SARS-CoV infection in the absence of CD4+and CD8+T cells and antibodies. Our findings provide new insights into the pathogenesis of SARS, demonstrating the important role of CD4+but not CD8+T cells in primary SARS-CoV infection in this model.
Wide-Scale Use of Notch Signaling Factor CSL/RBP-Jκ in RTA-Mediated Activation of Kaposi's Sarcoma-Associated Herpesvirus Lytic Genes
Authors: Persson Linda M. , Wilson Angus C.
Published: November 2009

For Kaposi's sarcoma-associated herpesvirus (KSHV; also called human herpesvirus 8 [HHV8]), the switch from latency to active lytic replication requires RTA, the product of open reading frame 50 (ORF50). RTA activates transcription from nearly 40 early and delayed-early viral promoters, mainly through interactions with cellular DNA binding proteins, such as CSL/RBP-Jκ, Oct-1, C/EBPα, and c-Jun. Reliance on cellular coregulators may allow KSHV to adjust its lytic program to suit different cellular contexts or interpret signals from the outside. CSL is a key component of the Notch signaling pathway and is targeted by several viruses. A search with known CSL binding sequences from cellular genes found at least 260 matches in the KSHV genome, many from regions containing known or suspected lytic promoters. Analysis of clustered sites located immediately upstream of ORF70 (thymidylate synthase), ORF19 (tegument protein), and ORF47 (glycoprotein L) uncovered RTA-responsive promoters that were validated using mRNAs isolated from KSHV-infected cells undergoing lytic reactivation. Notably, ORF19 behaves as a true late gene, indicating that RTA regulates all three phases of the lytic program. For each new promoter, the response to RTA was dependent on CSL, and 5 of the 10 candidate sites were shown to bind CSL
in vitro
. Analysis of individual sites highlighted the importance of a cytosine residue flanking the core CSL binding sequence. These findings broaden the role for CSL in coordinating the KSHV lytic gene expression program and help to define a signature motif for functional CSL sites within the viral genome.
Recombination, Reservoirs, and the Modular Spike: Mechanisms of Coronavirus Cross-Species Transmission
Authors: Graham Rachel L. , Baric Ralph S.
Published: November 2009

Over the past 30 years, several cross-species transmission events, as well as changes in virus tropism, have mediated significant animal and human diseases. Most notable is severe acute respiratory syndrome (SARS), a lower respiratory tract disease of humans that was first reported in late 2002 in Guangdong Province, China. The disease, which quickly spread worldwide over a period of 4 months spanning late 2002 and early 2003, infected over 8,000 individuals and killed nearly 800 before it was successfully contained by aggressive public health intervention strategies. A coronavirus (SARS-CoV) was identified as the etiological agent of SARS, and initial assessments determined that the virus crossed to human hosts from zoonotic reservoirs, including bats, Himalayan palm civets (Paguma larvata), and raccoon dogs (Nyctereutes procyonoides), sold in exotic animal markets in Guangdong Province. In this review, we discuss the molecular mechanisms that govern coronavirus cross-species transmission bothin vitroandin vivo, using the emergence of SARS-CoV as a model. We pay particular attention to how changes in the Spike attachment protein, both within and outside of the receptor binding domain, mediate the emergence of coronaviruses in new host populations.
Mutations in the Ectodomain of Newcastle Disease Virus Fusion Protein Confer a Hemagglutinin-Neuraminidase-Independent Phenotype
Authors: Ayllón Juan , Villar Enrique , Muñoz-Barroso Isabel
Published: November 2009

The entry of enveloped viruses into host cells is preceded by membrane fusion, which in paramyxoviruses is triggered by the fusion (F) protein. Refolding of the F protein from a metastable conformation to a highly stable postfusion form is critical for the promotion of fusion, although the mechanism is still not well understood. Here we examined the effects of mutations of individual residues of the F protein of Newcastle disease virus, located at critical regions of the protein, such as the C terminus of the N-terminal heptad repeat (HRA) and the N terminus of the C-terminal heptad repeat (HRB). Seven of the mutants were expressed at the cell surface, showing differences in antibody reactivity in comparison with the F wild type. The N211A, L461A, I463A, and I463F mutants showed a hyperfusogenic phenotype both in syncytium and in dye transfer assays. The four mutants promoted fusion more efficiently at lower temperatures than the wild type did, meaning they probably had lower energy requirements for activation. Moreover, the N211A, I463A, and I463F mutants exhibited hemagglutinin-neuraminidase (HN)-independent activity when influenza virus hemagglutinin (HA) was coexpressed as an attachment protein. The data are discussed in terms of alterations of the refolding pathway and/or the stability of the prefusion and fusion conformations.
Temperature Protects Insect Cells from Infection by Cricket Paralysis Virus
Authors: Cevallos Randal C. , Sarnow Peter
Published: November 2009

Heat shock is a well-known stress response characterized by a rapid synthesis of a set of proteins which are responsible for protection against stress. We examined the role of temperature on the growth of cricket paralysis virus, a member of the family
Dicistroviridae
, in insect cells. Heat shock caused an induction of heat shock protein-encoding mRNAs in uninfected cells but not in infected cells. While viral RNA and protein were abundant during heat shock, virion formation was inhibited at higher temperatures. The different susceptibility to pathogens at different temperatures is likely a crucial feature of host-pathogen interaction in cold-blooded animals.
Some Attenuated Variants of Vesicular Stomatitis Virus Show Enhanced Oncolytic Activity against Human Glioblastoma Cells relative to Normal Brain Cells
Authors: Wollmann Guido , Rogulin Vitaliy , Simon Ian , Rose John K. , van den Pol Anthony N.
Published: November 2009

Vesicular stomatitis virus (VSV) has been shown in laboratory studies to be effective against a variety of tumors, including malignant brain tumors. However, attenuation of VSV may be necessary to balance the potential toxicity toward normal cells, particularly when targeting brain tumors. Here we compared 10 recombinant VSV variants resulting from different attenuation strategies. Attenuations included gene shifting (VSV-p1-GFP/RFP), M protein mutation (VSV-M51), G protein cytoplasmic tail truncations (VSV-CT1/CT9), G protein deletions (VSV-dG-GFP/RFP), and combinations thereof (VSV-CT9-M51). Using
in vitro
viability and replication assays, the VSV variants were grouped into three categories, based on their antitumor activity and non-tumor-cell attenuation. In the first group, wild-type-based VSV-G/GFP, tumor-adapted VSV-rp30, and VSV-CT9 showed a strong antitumor profile but also retained some toxicity toward noncancer control cells. The second group, VSV-CT1, VSV-dG-GFP, and VSV-dG-RFP, had significantly diminished toxicity toward normal cells but showed little oncolytic action. The third group displayed a desired combination of diminished general toxicity and effective antitumor action; this group included VSV-M51, VSV-CT9-M51, VSV-p1-GFP, and VSV-p1-RFP. A member of the last group, VSV-p1-GFP, was then compared
in vivo
against wild-type-based VSV-G/GFP. Intranasal inoculation of young, postnatal day 16 mice with VSV-p1-GFP showed no adverse neurological effects, whereas VSV-G/GFP was associated with high lethality (80%). Using an intracranial tumor xenograft model, we further demonstrated that attenuated VSV-p1-GFP targets and kills human U87 glioblastoma cells after systemic application. We concluded that some, but not all, attenuated VSV mutants display a favorable oncolytic profile and merit further investigation.
Immune Evasion Proteins of Murine Cytomegalovirus Preferentially Affect Cell Surface Display of Recently Generated Peptide Presentation Complexes
Authors: Lemmermann Niels A. W. , Gergely Kerstin , Böhm Verena , Deegen Petra , Däubner Torsten , Reddehase Matthias J.
Published: November 2009

For recognition of infected cells by CD8 T cells, antigenic peptides are presented at the cell surface, bound to major histocompatibility complex class I (MHC-I) molecules. Downmodulation of cell surface MHC-I molecules is regarded as a hallmark function of cytomegalovirus-encoded immunoevasins. The molecular mechanisms by which immunoevasins interfere with the MHC-I pathway suggest, however, that this downmodulation may be secondary to an interruption of turnover replenishment and that hindrance of the vesicular transport of recently generated peptide-MHC (pMHC) complexes to the cell surface is the actual function of immunoevasins. Here we have used the model of murine cytomegalovirus (mCMV) infection to provide experimental evidence for this hypothesis. To quantitate pMHC complexes at the cell surface after infection in the presence and absence of immunoevasins, we generated the recombinant viruses mCMV-SIINFEKL and mCMV-Δm06m152-SIINFEKL, respectively, expressing the Kb-presented peptide SIINFEKL with early-phase kinetics in place of an immunodominant peptide of the viral carrier protein gp36.5/m164. The data revealed ∼10,000 Kbmolecules presenting SIINFEKL in the absence of immunoevasins, which is an occupancy of ∼10% of all cell surface Kbmolecules, whereas immunoevasins reduced this number to almost the detection limit. To selectively evaluate their effect on preexisting pMHC complexes, cells were exogenously loaded with SIINFEKL peptide shortly after infection with mCMV-SIINFEKA, in which endogenous presentation is prevented by an L174A mutation of the C-terminal MHC-I anchor residue. The data suggest that pMHC complexes present at the cell surface in advance of immunoevasin gene expression are downmodulated due to constitutive turnover in the absence of resupply.