bluuebunny/crossref_metadata_2025 huggingface dataset:
Highly Effective Serodiagnosis for Chagas' Disease
Authors: Hernández Pilar , Heimann Michael , Riera Cristina , Solano Marco , Santalla José , Luquetti Alejandro O. , Beck Ewald
Published: July 2010

Many proteins of
Trypanosoma cruzi
, the causative agent of Chagas' disease, contain characteristic arrays of highly repetitive immunogenic amino acid motifs. Diagnostic tests using these motifs in monomeric or dimeric form have proven to provide markedly improved specificity compared to conventional tests based on crude parasite extracts. However, in many cases the available tests still suffer from limited sensitivity. In this study we produced stable synthetic genes with maximal codon variability for the four diagnostic antigens, B13, CRA, TcD, and TcE, each containing between three and nine identical amino acid repeats. These genes were combined by linker sequences encoding short proline-rich peptides, giving rise to a 24-kDa fusion protein which was used as a novel diagnostic antigen in an enzyme-linked immunosorbent assay setup. Validation of the assay with a large number of well-characterized patient sera from Bolivia and Brazil revealed excellent diagnostic performance. The high sensitivity of the new test may allow future studies to use blood collected by finger prick and dried on filter paper, thus dramatically reducing the costs and effort for the detection of
T. cruzi
infection.
Bordetella holmesii
Bacteremia in Asplenic Children: Report of Four Cases Initially Misidentified as
Acinetobacter lwoffii
Authors: Panagopoulos Markos Ioannis , Saint Jean Maude , Brun Delphine , Guiso Nicole , Bekal Sadjia , Ovetchkine Philippe , Tapiero Bruce
Published: July 2010

Bordetella holmesii
is a fastidious Gram-negative rod that was initially identified in 1995. It causes bacteremia, predominantly among patients with anatomical or functional asplenia. We report four cases of
B. holmesii
bacteremia in asplenic children occurring within the last 4 years. In all cases,
B. holmesii
was misidentified by an automated system as
Acinetobacter lwoffii.
Evaluation of Matrix-Assisted Laser Desorption Ionization-Time of Flight Mass Spectrometry for Identification of Clinically Important Yeast Species
Authors: Stevenson Lindsay G. , Drake Steven K. , Shea Yvonne R. , Zelazny Adrian M. , Murray Patrick R.
Published: July 2010

We evaluated the use of matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) for the rapid identification of yeast species. Using Bruker Daltonics MALDI BioTyper software, we created a spectral database library with
m/z
ratios of 2,000 to 20,000 Da for 109 type and reference strains of yeast (44 species in 8 genera). The database was tested for accuracy by use of 194 clinical isolates (23 species in 6 genera). A total of 192 (99.0%) of the clinical isolates were identified accurately by MALDI-TOF MS. The MALDI-TOF MS-based method was found to be reproducible and accurate, with low consumable costs and minimal preparation time.
Characterization of
Staphylococcus aureus
Isolates with a Partial or Complete Absence of Staphylococcal Cassette Chromosome Elements
Authors: Wong Henry , Louie Lisa , Lo Reggie Y. C. , Simor Andrew E.
Published: July 2010

Detection of methicillin-resistant
Staphylococcus aureus
(MRSA) by single-locus PCR assays that target the extremity of the staphylococcal cassette chromosome-
mec
(SCC
mec
) and part of the adjacent
S. aureus
-specific open reading frame gene (
orfX
) is a significant diagnostic advancement, since it provides real-time detection directly from screening specimens. However, isolates harboring
mecA
deletions within SCC
mec
may result in false-positive identification of MRSA in these assays. We characterized 24 methicillin-susceptible
S. aureus
(MSSA) isolates that tested positive in one such assay to investigate this phenomenon. Seven isolates resembled USA100 and carried SCC
mec
II elements with
mecA
deletions that spanned 20 to 46 kbp. The
mecA
excisions in USA100-resembling isolates appeared to be linked with IS
431
transposable elements present in SCC
mec
II. For 17 isolates that resembled USA400 and/or MSSA476, the identity and possible excision of SCC elements could not be confirmed. The downstream common sequence (
dcs
) shared by SCC
mec
I, II, and IV elements was detected in these isolates. Sequence analysis of the chromosomal regions flanking the missing SCC element revealed an intact SCC integration site, a duplicate
dcs
, and the enterotoxin gene cluster downstream of
orfX
. An annealing sequence for one of the SCC
mec
-specific primers (mecii574) in the single-locus PCR assay was identified in the duplicate
dcs
. In the absence of SCC, a 176-bp amplicon can be generated from this mecii574 annealing sequence to yield a false-positive result. In conclusion, partial SCC
mec
II excisions via IS
431
elements in strains that resembled USA100 and the presence of a duplicate mecii574 annealing sequence in strains that resembled USA400/MSSA476 were identified as causes for false-positive results in a single-locus PCR assay that targets the SCC
mec
/
orfX
junction.
PCR-Single-Strand Conformational Polymorphism Method for Rapid Detection of Rifampin-Resistant
Mycobacterium tuberculosis
: Systematic Review and Meta-Analysis
Authors: Xu Hong-Bin , Jiang Rui-Hai , Sha Wei , Li Ling , Xiao He-Ping
Published: July 2010

The reference standard methods for drug susceptibility testing of
Mycobacterium tuberculosis
, such as culture on Lowenstein-Jensen or Middlebrook 7H10/11 medium, are very slow to give results; and due to the emergence of multidrug-resistant
M. tuberculosis
and extensively drug-resistant
M. tuberculosis
, there is an urgent demand for new, rapid, and accurate drug susceptibility testing methods. PCR-single-strand conformational polymorphism (PCR-SSCP) analysis has been proposed as a rapid method for the detection of resistance to rifampin, but its accuracy has not been systematically evaluated. We performed a systematic review and meta-analysis to evaluate the accuracy of PCR-SSCP analysis for the detection of rifampin-resistant tuberculosis. We searched the Medline, Embase, Web of Science, BIOSIS, and LILACS databases and contacted authors if additional information was required. Ten studies met our inclusion criteria for rifampin resistance detection. We applied the summary receiver operating characteristic (SROC) curve to perform the meta-analysis and to summarize diagnostic accuracy. The sensitivity of PCR-SSCP analysis for the rapid detection of rifampin-resistant tuberculosis was 0.79 (95% confidence interval [CI], 0.75 to 0.82), the specificity was 0.96 (95% CI, 0.94 to 0.98), the positive likelihood ratio was 16.10 (95% CI, 5.87 to 44.13), the negative likelihood ratio was 0.20 (95% CI, 0.10 to 0.40), and the diagnostic odds ratio was 100.93 (95% CI, 31.95 to 318.83). PCR-SSCP analysis is a sensitive and specific test for the rapid detection of rifampin-resistant
M. tuberculosis
. Additional studies in countries with a high prevalence of multidrug-resistant
M. tuberculosis
and also cost-effectiveness analysis are required in order to obtain a complete picture on the utility of this method for rapid drug resistance detection in
M. tuberculosis
.
Performance of the Abbott RealTi
m
e CT/NG for Detection of
Chlamydia trachomatis
and
Neisseria gonorrhoeae
Authors: Gaydos C. A. , Cartwright C. P. , Colaninno P. , Welsch J. , Holden J. , Ho S. Y. , Webb E. M. , Anderson C. , Bertuzis R. , Zhang L. , Miller T. , Leckie G. , Abravaya K. , Robinson J.
Published: July 2010

A multicenter clinical study was conducted to evaluate the performance characteristics of the Abbott RealTi
m
e CT/NG assay, a multiplex real-time PCR assay, for simultaneous detection of
Chlamydia trachomatis
and
Neisseria gonorrhoeae
. The specimens were collected from a total of 3,832 male and female subjects at 16 geographically diverse sites. Specimens included male and female urine samples, male urethral swabs, female endocervical swabs, and self-collected and clinician-collected vaginal swabs. Specimens were tested with the automated Abbott RealTi
m
e CT/NG assay, Aptima Combo 2 assay (Gen-Probe), ProbeTec ET CT/GC assay (Becton Dickinson), and culture for
N. gonorrhoeae
. The Aptima Combo 2 assay, the ProbeTec assay, and the
N. gonorrhoeae
culture were used as the reference assays. For each subject, a patient infected status (PIS) was determined based on the combined results from the reference assays. The overall prevalence in female subjects was 8.9% for
C. trachomatis
and 3.8% for
N. gonorrhoeae
. The overall male prevalence was 18.2% for
C. trachomatis
and 16.7% for
N. gonorrhoeae
. The overall sensitivity and specificity of the Abbott RealTi
m
e CT/NG assay were 92.4% and 99.2% for
C. trachomatis
and 96.9% and 99.7% for
N. gonorrhoeae
, respectively. In comparison, the sensitivity and specificity, respectively, for the Aptima Combo 2 assay were 94.5% and 99.0% for
C. trachomatis
and 96.1% and 99.5% for
N. gonorrhoeae
, and those for the ProbeTec ET assay were 90.3% and 99.5% for
C. trachomatis
and 92.0% and 97.3% for
N. gonorrhoeae
in this study. The Abbott RealTi
m
e CT/NG assay offers
C. trachomatis
and
N. gonorrhoeae
dual detection with high sensitivity and specificity. The automated assay provides a useful alternative nucleic acid amplification assay for clinical laboratories and clinicians.
Highly Sensitive and Quantitative Detection of the H274Y Oseltamivir Resistance Mutation in Seasonal A/H1N1 Influenza Virus
Authors: Operario Darwin J. , Moser Michael J. , St. George Kirsten
Published: July 2010

A C-to-T transition mutation in the neuraminidase gene from seasonal A/H1N1 causes a His-to-Tyr mutation at amino acid position 275 (H274Y, universal N2 numbering), conferring resistance against oseltamivir (Tamiflu). This mutation was first detected in clinical samples in Europe during the 2007-2008 influenza season. Viruses with this mutation reached a prevalence of ∼11% by the end of the season in North American isolates tested by the CDC. We developed a highly sensitive and specific quantitative real-time reverse transcriptase PCR assay to detect the H274Y mutation. This assay utilizes a 5′-methyl-isocytosine (isoC) residue and fluorescent reporters on genotype-specific primers. During PCR, a quencher coupled to isoguanine (isoG) is site-specifically incorporated complementary to the isoC/dye, resulting in loss of fluorescence. Optimization of primers and assay conditions produced a limit of detection of 100 gene copies per reaction for both wild-type and H274Y genotypes. In samples with mixed populations, it can reliably detect as little as a 1% wild-type or 0.1% H274Y component. This high sensitivity makes the assay usable on samples with viral loads too low for dideoxy or pyrosequencing analysis. Additionally, the assay distinguishes seasonal A/H1N1 from A/H3N2, influenza B, or 2009 pandemic A/H1N1, making it useful for influenza virus subtyping as well as for drug resistance detection. We probed seasonal A/H1N1 samples from the 2005-2006, 2006-2007, and 2007-2008 influenza seasons. Data from the new assay closely matched available drug resistance genotype data previously determined by dideoxy sequencing. The H274Y mutation was only found in samples from the 2007-2008 season.
A Multivalent Vaccination Strategy for the Prevention of Old World Arenavirus Infection in Humans
Authors: Botten Jason , Whitton J. Lindsay , Barrowman Polly , Sidney John , Whitmire Jason K. , Alexander Jeff , Kotturi Maya F. , Sette Alessandro , Buchmeier Michael J.
Published: July 2010

Arenaviruses cause severe human disease ranging from aseptic meningitis following lymphocytic choriomeningitis virus (LCMV) infection to hemorrhagic fever syndromes following infection with Guanarito virus (GTOV), Junin virus (JUNV), Lassa virus (LASV), Machupo virus (MACV), Sabia virus (SABV), or Whitewater Arroyo virus (WWAV). Cellular immunity, chiefly the CD8+T-cell response, plays a critical role in providing protective immunity following infection with the Old World arenaviruses LASV and LCMV. In the current study, we evaluated whether HLA class I-restricted epitopes that are cross-reactive among pathogenic arenaviruses could be identified for the purpose of developing an epitope-based vaccination approach that would cross-protect against multiple arenaviruses. We were able to identify a panel of HLA-A*0201-restricted peptides derived from the same region of the glycoprotein precursor (GPC) of LASV (GPC spanning residues 441 to 449 [GPC441-449]), LCMV (GPC447-455), JUNV (GPC429-437), MACV (GPC444-452), GTOV (GPC427-435), and WWAV (GPC428-436) that displayed high-affinity binding to HLA-A*0201 and were recognized by CD8+T cells in a cross-reactive manner following LCMV infection or peptide immunization of HLA-A*0201 transgenic mice. Immunization of HLA-A*0201 mice with the Old World peptide LASV GPC441-449or LCMV GPC447-455induced high-avidity CD8+T-cell responses that were able to kill syngeneic target cells pulsed with either LASV GPC441-449or LCMV GPC447-455in vivoand provided significant protection against viral challenge with LCMV. Through this study, we have demonstrated that HLA class I-restricted, cross-reactive epitopes exist among diverse arenaviruses and that individual epitopes can be utilized as effective vaccine determinants for multiple pathogenic arenaviruses.
Immunodominant HIV-Specific CD8+T-Cell Responses Are Common to Blood and Gastrointestinal Mucosa, and Gag-Specific Responses Dominate in Rectal Mucosa of HIV Controllers
Authors: Ferre April L. , Lemongello Donna , Hunt Peter W. , Morris Megan M. , Garcia Juan Carlos , Pollard Richard B. , Yee Hal F. , Martin Jeffrey N. , Deeks Steven G. , Shacklett Barbara L.
Published: July 2010

Previous studies have suggested that polyfunctional mucosal CD8+T-cell responses may be a correlate of protection in HIV controllers. Mucosal T-cell breadth and/or specificity may also contribute to defining protective responses. In this study, rectal CD8+T-cell responses to HIV Gag, Env, and Nef were mapped at the peptide level in four subject groups: elite controllers (n= 16; viral load [VL], <75 copies/ml), viremic controllers (n= 14; VL, 75 to 2,000 copies/ml), noncontrollers (n= 14; VL, >10,000 copies/ml), and antiretroviral-drug-treated subjects (n= 8; VL, <75 copies/ml). In all subject groups, immunodominant CD8+T-cell responses were generally shared by blood and mucosa, although there were exceptions. In HIV controllers, responses to HLA-B27- and HLA-B57-restricted epitopes were common to both tissues, and their magnitude (in spot-forming cells [SFC] per million) was significantly greater than those of responses restricted by other alleles. Furthermore, peptides recognized by T cells in both blood and rectal mucosa, termed “concordant,” elicited higher median numbers of SFC than discordant responses. In magnitude as well as breadth, HIV Gag-specific responses, particularly those targeting p24 and p7, dominated in controllers. Responses in noncontrollers were more evenly distributed among epitopes in Gag, Env, and Nef. Viremic controllers showed significantly broader mucosal Gag-specific responses than other groups. Taken together, these findings demonstrate that (i) Gag-specific responses dominate in mucosal tissues of HIV controllers; (ii) there is extensive overlap between CD8+T cells in blood and mucosal tissues, with responses to immunodominant epitopes generally shared by both sites; and (iii) mucosal T-cell response breadth alone cannot account for immune control.
Molecular Analysis of H7 Avian Influenza Viruses from Australia and New Zealand: Genetic Diversity and Relationships from 1976 to 2007
Authors: Bulach Dieter , Halpin Rebecca , Spiro David , Pomeroy Laura , Janies Daniel , Boyle David B.
Published: July 2010

Full-genome sequencing of 11 Australian and 1 New Zealand avian influenza A virus isolate (all subtype H7) has enabled comparison of the sequences of each of the genome segments to those of other subtype H7 avian influenza A viruses. The inference of phylogenetic relationships for each segment has been used to develop a model of the natural history of these viruses in Australia. Phylogenetic analysis of the hemagglutinin segment indicates that the Australian H7 isolates form a monophyletic clade. This pattern is consistent with the long-term, independent evolution that is, in this instance, associated with geographic regions. On the basis of the analysis of the other H7 hemagglutinin sequences, three other geographic regions for which similar monophyletic clades have been observed were confirmed. These regions are Eurasia plus Africa, North America, and South America. Analysis of the neuraminidase sequences from the H7N1, H7N3, and H7N7 genomes revealed the same region-based relationships. This pattern of independent evolution of Australian isolates is supported by the results of analysis of each of the six remaining genomic segments. These results, in conjunction with the occurrence of five different combinations of neuraminidase subtypes (H7N2, H7N3, H7N4, H7N6, H7N7) among the 11 Australian isolates, suggest that the maintenance host(s) is nearly exclusively associated with Australia. The single lineage of Australian H7 hemagglutinin sequences, despite the occurrence of multiple neuraminidase types, suggests the existence of a genetic pool from which a variety of reassortants arise rather than the presence of a small number of stable viral clones. This pattern of evolution is likely to occur in each of the regions mentioned above.
Interplay of Acute and Persistent Infections Caused by Venezuelan Equine Encephalitis Virus Encoding Mutated Capsid Protein
Authors: Atasheva Svetlana , Krendelchtchikova Valentina , Liopo Anton , Frolova Elena , Frolov Ilya
Published: July 2010

Venezuelan equine encephalitis virus (VEEV) is a significant human and animal pathogen. The highlight of VEEV replication
in vitro
, in cells of vertebrate origin, is the rapid development of cytopathic effect (CPE), which is strongly dependent upon the expression of viral capsid protein. Besides being an integral part of virions, the latter protein is capable of (i) binding both the nuclear import and nuclear export receptors, (ii) accumulating in the nuclear pore complexes, (iii) inhibiting nucleocytoplasmic trafficking, and (iv) inhibiting transcription of cellular ribosomal and messenger RNAs. Using our knowledge of the mechanism of VEEV capsid protein function in these processes, we designed VEEV variants containing combinations of mutations in the capsid-coding sequences. These mutations made VEEV dramatically less cytopathic but had no effect on infectious virus production. In cell lines that have defects in type I interferon (IFN) signaling, the capsid mutants demonstrated very efficient persistent replication. In other cells, which have no defects in IFN production or signaling, the same mutants were capable of inducing a long-term antiviral state, downregulating virus replication to an almost undetectable level. However, ultimately, these cells also developed a persistent infection, characterized by continuous virus replication and beta IFN (IFN-β) release. The results of this study demonstrate that the long-term cellular antiviral state is determined by the synergistic effects of type I IFN signaling and the antiviral reaction induced by replicating viral RNA and/or the expression of VEEV-specific proteins. The designed mutants represent an important model for studying the mechanisms of cell interference with VEEV replication and development of persistent infection.
9-Borafluorenes – NMR spectroscopy and DFT calculations. Molecular structure of 1,2-(2,2′-diphenylylene)-1,2-diethyldiborane
Authors: Wrackmeyer Bernd , Thoma Peter , Kempe Rhett , Glatz Germund
Published: July 2010

9-Borafluorene derivatives 1 (9-R = Et (a), Ph (b), Cl (c), NEt2 (d)), the pyridine adduct 1py+ and 1,2-(2,2′-biphenylylene)-1,2-diethyldiborane(6) (3), were studied by 11B and 13C NMR spectroscopy to obtain a fairly complete data set for the first time. The molecular structure of the doubly hydrogen-bridged 1,2-diphenylenediborane 3 was determined by X-ray diffraction. The gas-phase structures of the compounds 1, related derivatives, and of some doubly hydrogen-bridged 1,2-diphenylenediboranes were optimized by quantum chemical calculations (B3LYP/6-311+G(d,p) level of theory) and NMR parameters, such as chemical shifts, 11B chemical shift tensors and indirect nuclear 13C–11B spin–spin coupling constants were calculated at the same level of theory and compared with experimental data.
INVESTIGATION PROPERTIES OF a-Si1-xCx:H FILMS ELABORATED BY CO-SPUTTERING OF Si AND 6H-SiC
Authors: KEFFOUS AISSA , CHERIET ABDELHAK , BELKACEM YOUCEF , MANSERI AMAR , GABOUZE NOUREDDINE , KECHOUANE MOHAMED , BRIGHET AMER , BOUKEZZATA ASSIA , KACI SAMIRA , MENOUS ISA , NEZZAL GHANIA , GUERBOUS LAKHDAR , MENARI HAMID
Published: July 2010

Hydrogenated amorphous SiC films ( a - Si 1-x C x: H ) were prepared by DC magnetron sputtering technique on p type Si (100) and corning 9075 substrates at low temperature, by using 32 sprigs of silicon carbide (6 H - SiC ). The deposited film a - Si 1-x C x: H was realized under a mixture of argon and hydrogen gases. The ( a - Si 1-x C x: H ) films have been investigated by scanning electronic microscopy equipped with EDS system (SEM-EDS), X-rays diffraction (XRD), secondary ions mass spectrometry (SIMS), Fourier transform infrared spectroscopy (FTIR), UV-visible-IR spectrophotometry, and photoluminescence (PL). XRD results showed that the deposited film was amorphous with a structure of a - Si 0.81 C 0.19: H corresponding to 19 at.% carbon. The photoluminescence response of the samples was observed in the visible range at room temperature with two peaks centered at 463 nm (2.68 eV) and 542 nm (2.29 eV). The structural properties and the origin of the luminescence were discussed.
Fermi-Hubbard Physics with Atoms in an Optical Lattice
Authors: Esslinger Tilman
Published: July 2010

The Fermi-Hubbard model is a key concept in condensed matter physics and provides crucial insights into electronic and magnetic properties of materials. Yet, the intricate nature of Fermi systems poses a barrier to answering important questions concerning d-wave superconductivity and quantum magnetism. Recently, it has become possible to experimentally realize the Fermi-Hubbard model using a fermionic quantum gas loaded into an optical lattice. In this atomic approach to the Fermi-Hubbard model, the Hamiltonian is a direct result of the optical lattice potential created by interfering laser fields and short-ranged ultracold collisions. It provides a route to simulate the physics of the Hamiltonian and to address open questions and novel challenges of the underlying many-body system. This review gives an overview of the current efforts in understanding and realizing experiments with fermionic atoms in optical lattices and discusses key experiments in the metallic, band-insulating, superfluid, and Mott-insulating regimes.
Rheology of Soft Materials
Authors: Chen Daniel T.N. , Wen Qi , Janmey Paul A. , Crocker John C. , Yodh Arjun G.
Published: July 2010

Research on soft materials, including colloidal suspensions, glasses, pastes, emulsions, foams, polymer networks, liquid crystals, granular materials, and cells, has captured the interest of scientists and engineers in fields ranging from physics and chemical engineering to materials science and cell biology. Recent advances in rheological methods to probe mechanical responses of these complex media have been instrumental for producing new understanding of soft matter and for generating novel technological applications. This review surveys these technical developments and current work in the field, with partial aim to illustrate open questions for future research.
Short-term voluntary exercise in the rat causes bone modeling without initiating a physiological stress response
Authors: Rosa Brielle V. , Firth Elwyn C. , Blair Hugh T. , Vickers Mark H. , Morel Patrick C. H. , Cockrem John F.
Published: July 2010

Recent research has revealed a neuroendocrine connection between the skeleton and metabolism. Exercise alters both bone modeling and energy balance and may be useful in further developing our understanding of this complex interplay. However, research in this field requires an animal model of exercise that does not cause a physiological stress response in the exercised subjects. In this study, we develop a model of short-term voluntary exercise in the female rat that causes bone modeling without causing stress. Rats were randomly assigned to one of three age-matched groups: control, tower climbing, and squat exercise (rising to an erect bipedal stance). Exercise for 21 days resulted in bone modeling as assessed by peripheral quantitative computed tomography. Fecal corticosterone output was used to assess physiological stress at three time points during the study (preexercise, early exercise, and late in the exercise period). There were no differences in fecal corticosterone levels between groups or time points. This model of voluntary exercise in the rat will be useful for future studies of the influence of exercise on the relationship between skeletal and metabolic health and may be appropriate for investigation of the developmental origins of those effects.
Structural neuroplasticity following T5 spinal cord transection: increased cardiac sympathetic innervation density and SPN arborization
Authors: Lujan Heidi L. , Palani Gurunanthan , DiCarlo Stephen E.
Published: July 2010

When the spinal cord is injured at or below thoracic level 5 (T5), cardiovascular control is markedly unbalanced as the heart and blood vessels innervated by upper thoracic segments remain under brain stem control, whereas the vasculature of the lower body is affected by unregulated spinal reflexes. Importantly, the regulation of heart rate and cardiac function is abnormal after spinal cord injury (SCI) at T5 because sympathetic outflow to the heart is increased. An increase in tonic sympathetic outflow may be attributable to multiple mechanisms, such as increases in cardiac sympathetic innervation density, altered morphology of stellate ganglia neurons, and/or structural neuroplasticity of cardiac sympathetic preganglionic neurons (SPNs). Furthermore, these neuroplastic changes associated with SCI may be mediated by nerve growth factor (NGF). NGF is a neurotrophin that supports the survival and differentiation of sympathetic neurons and enhances target innervation. Therefore, we tested the hypothesis that T5 spinal cord transection (T5X) is associated with an increased left ventricular (LV) NGF content, LV sympathetic innervation density, and cardiac SPN arborization. In intact and paraplegic (9 wk posttransection) rats, LV NGF content (ELISA), LV sympathetic innervation density (tyrosine hydroxylase immunohistochemistry), and cardiac SPN arborization (cholera toxin B immunohistochemistry and Sholl Analysis) were determined. Paraplegia, compared with intact, significantly increased LV NGF content, LV sympathetic innervation density, and cardiac SPN arborization. Thus, altered autonomic behavior following SCI is associated with structural neuroplastic modifications.
Molecular conformation and filtration properties of anionic Ficoll
Authors: Groszek Joseph , Li Lingyan , Ferrell Nicholas , Smith Ross , Zorman Christian A. , Hofmann Christina L. , Roy Shuvo , Fissell William H.
Published: July 2010

The physiology of glomerular permselectivity remains mechanistically obscure, despite its importance in human disease. Although electrical contributions to glomerular permselectivity have long been considered important, two recent reports demonstrated enhanced glomerular permeability to anionic versus neutral polysaccharides. The interpretation of these observations is complicated by confounding of the effects of chemical modification on charge with effects on size and shape. In this report, neutral and anionic Ficoll are characterized by size-exclusion chromatography with online light scattering and viscometry and filtration through a highly defined anionic filtration membrane. Neutral and carboxymethylated Ficoll are nearly identical in size and conformation, yet carboxymethylated Ficoll is retained by an anionic membrane in excess of neutral Ficoll. This suggests that comparisons between clearances of neutral and carboxymethylated Ficoll may be a sensitive probe of electrostatic interactions independent of size and conformation.
The Effect of Bilateral Isometric Forces in Different Directions on Motor Cortical Function in Humans
Authors: Yedimenko Juliette A. , Perez Monica A.
Published: July 2010

The activity in the primary motor cortex (M1) reflects the direction of movements, but little is known about physiological changes in the M1 during generation of bilateral isometric forces in different directions. Here, we used transcranial magnetic stimulation to examine motor evoked potentials (MEPs), short-interval intracortical inhibition (SICI), and interhemispheric inhibition (IHI) in the left first dorsal interosseous (FDI) during isometric index finger abduction while the right index finger remained at rest or performed isometric forces in different directions (abduction or adduction) and in different postures (prone and supine). Left FDI MEPs were suppressed during bilateral compared with unilateral forces, with a stronger suppression when the right index finger force was exerted in the adduction direction regardless of hand posture. IHI targeting the left FDI increased during bilateral compared with unilateral forces and this increase was stronger during right index finger adduction despite the posture of the right hand. SICI decreased to a similar extent during both bilateral forces in both hand postures. Thus generation of index finger isometric forces away from the body midline (adduction direction), regardless of the muscle engaged in the task, down-regulates corticospinal output in the contralateral active hand to a greater extent than forces exerted toward the body midline (abduction direction). Transcallosal inhibition, but not GABAergic intracortical circuits, was modulated by the direction of the force. These findings suggest that during generation of bimanual isometric forces the M1 is driven by “extrinsic” parameters related to the hand action.
Sexual Assault: A Report on Human Immunodeficiency Virus Postexposure Prophylaxis
Authors: Griffith William F. , Ackerman Gary E. , Zoellner Cindy L. , Sheffield Jeanne S.
Published: July 2010

The objective of this report is to describe an urban county hospital human immunodeficiency virus (HIV) infection prevention protocol offering prophylactic combination antiretroviral medications to female victims of sexual assault. A retrospective chart review was conducted from June, 2007 through June, 2008 of 151 women who were prescribed antiretroviral prophylaxis by protocol. All women receiving HIV prophylaxis initially screened HIV seronegative. Of the 58 women who reported taking any HIV prophylaxis, 36 (62%) were HIV screened at 12 and/or 24 weeks and none had HIV seroconverted. Although the initiation of an HIV post exposure prophylaxis protocol for sexual assault in a county hospital population is feasible, patient follow‐up for counseling and HIV serostatus evaluation is an identified barrier
Tumor-Stromal Interactions Influence Radiation Sensitivity in Epithelial- versus Mesenchymal-Like Prostate Cancer Cells
Authors: Josson Sajni , Sharp Starlette , Sung Shian-Ying , Johnstone Peter A. S. , Aneja Ritu , Wang Ruoxiang , Gururajan Murali , Turner Timothy , Chung Leland W. K. , Yates Clayton
Published: July 2010

HS-27a human bone stromal cells, in 2D or 3D coultures, induced cellular plasticity in human prostate cancerARCaPEandARCaPMcells in an EMT model. CoculturedARCaPEorARCaPMcells with HS-27a, developed increased colony forming capacity and growth advantage, withARCaPEexhibiting the most significant increases in presence of bone or prostate stroma cells. Prostate (Pt-N or Pt-C) or bone (HS-27a) stromal cells induced significant resistance to radiation treatment inARCaPEcells compared toARCaPMcells. However pretreatment with anti-E-cadherin antibody (SHEP8-7) or anti-alpha v integrin blocking antibody (CNT095) significantly decreased stromal cell-induced radiation resistance in bothARCaPE- andARCaPM-cocultured cells. Taken together the data suggest that mesenchymal-like cancer cells reverting to epithelial-like cells in the bone microenvironment through interaction with bone marrow stromal cells and reexpress E-cadherin. These cell adhesion molecules such as E-cadherin and integrin alpha v in cancer cells induce cell survival signals and mediate resistance to cancer treatments such as radiation.
Hysterectomy—Current Methods and Alternatives for Benign Indications
Authors: Papadopoulos Michail S. , Tolikas Athanasios C. , Miliaras Dimosthenis E.
Published: July 2010

Hysterectomy is the commonest gynecologic operation performed not only for malignant disease but also for many benign conditions such as fibroids, endometrial hyperplasia, adenomyosis, uterine prolapse, dysfunctional uterine bleeding, and cervical intraepithelial neoplasia. There are many approaches to hysterectomy for benign disease: abdominal hysterectomy, vaginal hysterectomy, laparoscopic assisted vaginal hysterectomy (LAVH) where a vaginal hysterectomy is assisted by laparoscopic procedures that do not include uterine artery ligation, total laparoscopic hysterectomy (TLH) where the laparoscopic procedures include uterine artery ligation, and subtotal laparoscopic hysterectomy (STLH) where there is no vaginal component and the uterine body is removed using a morcelator. In the last decades, many new techniques, alternative to hysterectomy with conservation of the uterus have been developed. They use modern technologies and their results are promising and in many cases comparable with hysterectomy. This paper is a review of all the existing hysterectomy techniques and the alternative methods for benign indications.
Robust Reliable Stabilization of Switched Nonlinear Systems with Time‐Varying Delays and Delayed Switching
Authors: Xiang Zhengrong , Chen Qingwei
Published: July 2010

This paper is concerned with the problem of robust reliable stabilization of switched nonlinear systems with time‐varying delays and delayed switching is investigated. The parameter uncertainties are allowed to be norm‐bounded. The switching instants of the controller experience delays with respect to those of the system. The purpose of this problem is to design a reliable state feedback controller such that, for all admissible parameter uncertainties and actuator failure, the system state of the closed‐loop system is exponentially stable. We show that the addressed problem can be solved by means of algebraic matrix inequalities. The explicit expression of the desired robust controllers is derived in terms of linear matrix inequalities (LMIs).
Analysis of Control Structure for Turning Maneuvers
Authors: Shih Chia-Hung , Yamamura Saburo , Chen Chen-Yuan
Published: July 2010

Although there have been a variety of studies about vehicular maneuvers on land or sea, maneuvers within a small area that require direction changes have rarely been discussed. The arrival or departure of ships to or from ports requires safe and effective turning maneuvers within a narrow area. In this study, we decompose the maneuver procedure into several operations, then establish an operational model, simplifying the behaviors of rudders and thrusters that operate the turning maneuver, ignoring the environmental factors. This proposed model is a unified model that can be applied to all types of ships, regardless of individual characteristics. Each operation is defined as an isolated procedure that has its own effect on the maneuver behavior and the whole maneuver procedure can be treated as a combination of these operations. A case study using this model to construct the optimal turning maneuver within a limited sea area is also provided.
Nanofibers Offer Alternative Ways to the Treatment of Skin Infections
Authors: Heunis T. D. J. , Dicks L. M. T.
Published: July 2010

Injury to the skin causes a breach in the protective layer surrounding the body. Many pathogens are resistant to antibiotics, rendering conventional treatment less effective. This led to the use of alternative antimicrobial compounds, such as silver ions, in skin treatment. In this review nanofibers, and the incorporation of natural antimicrobial compounds in these scaffolds, are discussed as an alternative way to control skin infections. Electrospinning as a technique to prepare nanofibers is discussed. The possibility of using these structures as drug delivery systems is investigated.
MET Receptor Sequence Variants R970C and T992I Lack Transforming Capacity
Authors: Tyner Jeffrey W. , Fletcher Luke B. , Wang Ellen Q. , Yang Wayne F. , Rutenberg-Schoenberg Michael L. , Beadling Carol , Mori Motomi , Heinrich Michael C. , Deininger Michael W. , Druker Brian J. , Loriaux Marc M.
Published: July 2010

High-throughput sequencing promises to accelerate the discovery of sequence variants, but distinguishing oncogenic mutations from irrelevant “passenger” mutations remains a major challenge. Here we present an analysis of two sequence variants of the MET receptor (hepatocyte growth factor receptor) R970C and T992I (also designated R988C and T1010I). Previous reports indicated that these sequence variants are transforming and contribute to oncogenesis. We screened patients with chronic lymphocytic leukemia (CLL), acute myeloid leukemia (AML), chronic myelomonocytic leukemia (CMML), colorectal cancer, endometrial cancer, thyroid cancer, or melanoma, as well as individuals without cancer, and found these variants at low frequencies in most cohorts, including normal individuals. No evidence of increased phosphorylation or transformative capacity by either sequence variant was found. Because small-molecule inhibitors for MET are currently in development, it will be important to distinguish between oncogenic sequence variants and rare single-nucleotide polymorphisms to avoid the use of unnecessary, and potentially toxic, cancer therapy agents. Cancer Res; 70(15); 6233–7. ©2010 AACR.
A Novel FRET-Based Biosensor for the Measurement of BCR-ABL Activity and Its Response to Drugs in Living Cells
Authors: Mizutani Tatsuaki , Kondo Takeshi , Darmanin Stephanie , Tsuda Masumi , Tanaka Shinya , Tobiume Minoru , Asaka Masahiro , Ohba Yusuke
Published: July 2010

Purpose: To develop a novel diagnostic method for the assessment of drug efficacy in chronic myeloid leukemia (CML) patients individually, we generated a biosensor that enables the evaluation of BCR-ABL kinase activity in living cells using the principle of fluorescence resonance energy transfer (FRET).

Experimental Design: To develop FRET-based biosensors, we used CrkL, the most characteristic substrate of BCR-ABL, and designed a protein in which CrkL is sandwiched between Venus, a variant of YFP, and enhanced cyan fluorescent protein, so that CrkL intramolecular binding of the SH2 domain to phosphorylated tyrosine (Y207) increases FRET efficiency. After evaluation of the properties of this biosensor by comparison with established methods including Western blotting and flow cytometry, BCR-ABL activity and its response to drugs were examined in CML patient cells.

Results: After optimization, we obtained a biosensor that possesses higher sensitivity than that of established techniques with respect to measuring BCR-ABL activity and its suppression by imatinib. Thanks to its high sensitivity, this biosensor accurately gauges BCR-ABL activity in relatively small cell numbers and can also detect <1% minor drug-resistant populations within heterogeneous ones. We also noticed that this method enabled us to predict future onset of drug resistance as well as to monitor the disease status during imatinib therapy, using patient cells.

Conclusion: In consideration of its quick and practical nature, this method is potentially a promising tool for the prediction of both current and future therapeutic responses in individual CML patients, which will be surely beneficial for both patients and clinicians. Clin Cancer Res; 16(15); 3964–75. ©2010 AACR.
CD8+ Enriched “Young” Tumor Infiltrating Lymphocytes Can Mediate Regression of Metastatic Melanoma
Authors: Dudley Mark E. , Gross Colin A. , Langhan Michelle M. , Garcia Marcos R. , Sherry Richard M. , Yang James C. , Phan Giao Q. , Kammula Udai S. , Hughes Marybeth S. , Citrin Deborah E. , Restifo Nicholas P. , Wunderlich John R. , Prieto Peter A. , Hong Jenny J. , Langan Russell C. , Zlott Daniel A. , Morton Kathleen E. , White Donald E. , Laurencot Carolyn M. , Rosenberg Steven A.
Published: July 2010

Purpose: Tumor-infiltrating lymphocytes (TIL) and interleukin (IL)-2 administered following lymphodepletion can cause the durable complete regression of bulky metastatic melanoma in patients refractory to approved treatments. However, the generation of a unique tumor-reactive TIL culture for each patient may be prohibitively difficult. We therefore investigated the clinical and immunologic impact of unscreened, CD8+ enriched “young” TIL.

Experimental Design: Methods were developed for generating TIL that minimized the time in culture and eliminated the individualized tumor-reactivity screening step. Thirty-three patients were treated with these CD8+ enriched young TIL and IL-2 following nonmyeloablative lymphodepletion (NMA). Twenty-three additional patients were treated with CD8+ enriched young TIL and IL-2 after lymphodepletion with NMA and 6 Gy of total body irradiation.

Results: Young TIL cultures for therapy were successfully established from 83% of 122 consecutive melanoma patients. Nineteen of 33 patients (58%) treated with CD8+ enriched young TIL and NMA had an objective response (Response Evaluation Criteria in Solid Tumors) including 3 complete responders. Eleven of 23 patients (48%) treated with TIL and 6 Gy total body irradiation had an objective response including 2 complete responders. At 1 month after TIL infusion the absolute CD8+ cell numbers in the periphery were highly correlated with response.

Conclusions: This study shows that a rapid and simplified method can be used to reliably generate CD8+ enriched young TIL for administration as an individualized therapy for advanced melanoma, and may allow this potentially effective treatment to be applied at other institutions and to reach additional patients. Clin Cancer Res; 16(24); 6122–31. ©2010 AACR.
Fluorescence Resonance Energy Transfer Biosensors for Cancer Detection and Evaluation of Drug Efficacy
Authors: Lu Shaoying , Wang Yingxiao
Published: July 2010

A sensitive and specific fluorescence resonance energy transfer (FRET) biosensor was developed by Mizutani and colleagues and applied to detect the activity of BCR-ABL kinase in live cells. This biosensor allowed the detection of cancerous and drug-resistant cells, and the evaluation of kinase inhibitor efficacy. Future biosensor development and imaging can increasingly contribute to cancer diagnosis and therapeutics. Clin Cancer Res; 16(15); 3822–4. ©2010 AACR.
PIM Kinase Inhibitors Downregulate STAT3Tyr705 Phosphorylation
Authors: Chang Marisa , Kanwar Nisha , Feng Eric , Siu Allan , Liu Xiujie , Ma Dawei , Jongstra Jan
Published: July 2010

Using a cell-based high-throughput screen designed to detect small chemical compounds that inhibit cell growth and survival, we identified three structurally related compounds, 21A8, 21H7, and 65D4, with differential activity on cancer versus normal cells. Introduction of structural modifications yielded compound M-110, which inhibits the proliferation of prostate cancer cell lines with IC50s of 0.6 to 0.9 μmol/L, with no activity on normal human peripheral blood mononuclear cells up to 40 μmol/L. Screening of 261 recombinant kinases and subsequent analysis revealed that M-110 is a selective inhibitor of the PIM kinase family, with preference for PIM-3. The prostate cancer cell line DU-145 and the pancreatic cancer cell line MiaPaCa2 constitutively express activated STAT3 (pSTAT3Tyr705). Treatment of DU-145 cells with M-110 or with a structurally unrelated PIM inhibitor, SGI-1776, significantly reduces pSTAT3Tyr705 expression without affecting the expression of STAT3. Furthermore, treatment of DU-145 cells with M-110 attenuates the interleukin-6–induced increase in pSTAT3Tyr705. To determine which of the three PIM kinases is most likely to inhibit expression of pSTAT3Tyr705, we used PIM-1–, PIM-2–, or PIM-3–specific siRNA and showed that knockdown of PIM-3, but not of PIM-1 or PIM-2, in DU-145 cells results in a significant downregulation of pSTAT3Tyr705. The phosphorylation of STAT5 on Tyr694 in 22Rv1 cells is not affected by M-110 or SGI-1776, suggesting specificity for pSTAT3Tyr705. These results identify a novel role for PIM-3 kinase as a positive regulator of STAT3 signaling and suggest that PIM-3 inhibitors cause growth inhibition of cancer cells by downregulating the expression of pSTAT3Tyr705. Mol Cancer Ther; 9(9); 2478–87. ©2010 AACR.
Role of Perfusion CT in Assessing Tumor Blood Flow and Malignancy Level of Gastric Cancer
Authors: Satoh Asami , Shuto Kiyohiko , Okazumi Shinichi , Ohira Gaku , Natsume Toshiyuki , Hayano Koichi , Narushima Kazuo , Saito Hiroshige , Ohta Takumi , Nabeya Yoshihiro , Yanagawa Noriyuki , Matsubara Hisahiro
Published: July 2010

Background/Aims: Intratumoral hemodynamics or tumor perfusion is useful in understanding the pathological background of the cancer. A parameter for a non-invasive, preoperative assessment of tumor perfusion has yet to be developed. Methods: The study included 50 patients who underwent surgery for gastric cancer. Perfusion computed tomography (P-CT) was performed using a 16-row multidetector CT, and tumor blood flow (ml/min/100 g tissue) values were measured. We compared blood flow with histopathological characteristics and evaluated its correlation with microvessel density and tumor stromal density and calculated the ratio of vessels and stromal tissue. Results: There was a significant decrease in blood flow in advanced tumor depth, peritoneal dissemination and undifferentiated subtypes. Cases with Lauren’s diffuse type carcinoma were found to have decreased blood flow compared to the mixed or intestinal type. As for the stromal structure, despite the lack of correlation with microvessel density, blood flow significantly decreased with increased stromal density. Conclusions: Decreased blood flow value acquired from P-CT may reflect a progressive state of gastric cancer. The pathological background for this relation involves the tumor stroma. Tumor perfusion decreased as the stage and malignant character of the tumor advanced, and therefore P-CT could be a better strategy to estimate the malignancy level of cancer.