bluuebunny/crossref_metadata_2025 huggingface dataset:
Evaluation of the Antigenic Diversity of Placenta-Binding
Plasmodium falciparum
Variants and the Antibody Repertoire among Pregnant Women
Authors: Hommel Mirja , Elliott Salenna R. , Soma Viju , Kelly Greg , Fowkes Freya J. I. , Chesson Joanne M. , Duffy Michael F. , Bockhorst Joseph , Avril Marion , Mueller Ivo , Raiko Andrew , Stanisic Danielle I. , Rogerson Stephen J. , Smith Joseph D. , Beeson James G.
Published: February 2010

Pregnant women are infected by specific variants of
Plasmodium falciparum
that adhere and accumulate in the placenta. Using serological and molecular approaches, we assessed the global antigenic diversity of surface antigens expressed by placenta-binding isolates to better understand immunity to malaria in pregnancy and evolution of polymorphisms and to inform vaccine development. We found that placenta-binding isolates originating from all major regions where malaria occurs were commonly recognized by antibodies in different populations of pregnant women. There was substantial antigenic overlap and sharing of epitopes between isolates, including isolates from distant geographic locations, suggesting that there are limitations to antigenic diversity; however, differences between populations and isolates were also seen. Many women had cross-reactive antibodies and/or a broad repertoire of antibodies to different isolates. Studying VAR2CSA as the major antigen expressed by placenta-binding isolates, we identified antibody epitopes encoded by variable sequence blocks in the DBL3 domain. Analysis of global
var2csa
DBL3 sequences demonstrated that there was extensive sharing of variable blocks between Africa, Asia, Papua New Guinea, and Latin America, which likely contributes to the high level of antigenic overlap between different isolates. However, there was also evidence of geographic clustering of sequences and differences in VAR2CSA sequences between populations. The results indicate that there is limited antigenic diversity in placenta-binding isolates and may explain why immunity to malaria in pregnancy can be achieved after exposure during one pregnancy. Inclusion of a limited number of variants in a candidate vaccine may be sufficient for broad population coverage, but geographic considerations may also have to be included in vaccine design.
Linking endoplasmic reticulum stress to cell death in hepatocytes: roles of C/EBP homologous protein and chemical chaperones in palmitate-mediated cell death
Authors: Pfaffenbach Kyle T. , Gentile Christopher L. , Nivala Angela M. , Wang Dong , Wei Yuren , Pagliassotti Michael J.
Published: February 2010

Prolonged endoplasmic reticulum (ER) stress and activation of the unfolded protein response (UPR) have been linked to apoptosis via several mechanisms, including increased expression of C/EBP homologous protein ( Chop). Increased long-chain fatty acids, in particular saturated fatty acids, induce ER stress, Chop expression, and apoptosis in liver cells. The first aim of the present study was to determine the role of Chop in lipid-induced hepatocyte cell death and liver injury induced by a methionine-choline-deficient diet. Albumin-bound palmitate increased Chop gene and protein expression in a dose-dependent fashion in H4IIE liver cells. siRNA-mediated silencing of Chop in H4IIE liver cells reduced thapsigargin-mediated cell death by ∼40% and delayed palmitate-mediated cell death, but only at high concentrations of palmitate (400–500 μM). Similar results were observed in primary hepatocytes isolated from Chop-knockout mice. Indices of liver injury were also not reduced in Chop-knockout mice provided a methionine-choline-deficient diet. To ascertain whether ER stress was linked to palmitate-induced cell death, primary hepatocytes were incubated in the absence or presence of the chemical chaperones taurine-conjugated ursodeoxycholic acid or 4-phenylbutyric acid. The presence of either of these chemical chaperones protected liver cells from palmitate-mediated ER stress and cell death, in part, via inhibition of JNK activation. These data suggest that ER stress is linked to palmitate-mediated cell death via mechanisms that include JNK activation.
Nitration of specific tyrosines in FoF1ATP synthase and activity loss in aging
Authors: Haynes Virginia , Traaseth Nathaniel J. , Elfering Sarah , Fujisawa Yasuko , Giulivi Cecilia
Published: February 2010

It has been reported that C-nitration of proteins occurs under nitrative/oxidative stress; however, its role in pathophysiological situations is not fully understood. In this study, we determined that nitration of Tyr345and Tyr368in the β-subunit of the mitochondrial FoF1-ATPase is a major target for nitrative stress in rat liver under in vivo conditions. The chemical characteristics of these Tyr make them suitable for a facilitated nitration (solvent accessibility, consensus sequence, and p Ka). Moreover, β-subunit nitration increased significantly with the age of the rats (from 4 to 80 weeks old) and correlated with decreased ATP hydrolysis and synthesis rates. Although its affinity for ATP binding was unchanged, maximal ATPase activity decreased between young and old rats by a factor of two. These changes directly impacted the available ATP concentration in vivo, and it was expected that they would affect multiple cellular ATP-dependent processes. For instance, at least 50% of available [ATP] in the liver of older rats would have to be committed to sustain maximal Na+-K+-ATPase activity, whereas only 30% would be required for young rats. If this requirement was not fulfilled, the osmoregulation and Na+-nutrient cotransport in liver of older rats would be compromised. On the basis of our studies, we propose that targeted nitration of the β-subunit is an early marker for nitrative stress and aging.
Enhancement of Synchronization Between Hippocampal and Amygdala Theta Waves Associated With Pontine Wave Density
Authors: Karashima Akihiro , Katayama Norihiro , Nakao Mitsuyuki
Published: February 2010

Theta waves in the amygdala are known to be synchronized with theta waves in the hippocampus. Synchronization between amygdala and hippocampal theta waves is considered important for neuronal communication between these regions during the memory-retrieval process. These theta waves are also observed during rapid eye movement (REM) sleep. However, few studies have examined the mechanisms and functions of theta waves during REM sleep. This study examined correlations between the dynamics of hippocampal and amygdala theta waves and pontine (P) waves in the subcoeruleus region, which activates many brain areas including the hippocampus and amygdala, during REM sleep in rats. We confirmed that the frequency of hippocampal theta waves increased in association with P wave density, as shown in our previous study. The frequency of amygdala theta waves also increased with in associated with P wave density. In addition, we confirmed synchronization between hippocampal and amygdala theta waves during REM sleep in terms of the cross-correlation function and found that this synchronization was enhanced in association with increased P wave density. We further studied theta wave synchronization associated with P wave density by lesioning the pontine subcoeruleus region. This lesion not only decreased hippocampal and amygdala theta frequency, but also degraded theta wave synchronization. These results indicate that P waves enhance synchronization between regional theta waves. Because hippocampal and amygdala theta waves and P waves are known to be involved in learning and memory processes, these results may help clarify these functions during REM sleep.
Size of Error Affects Cerebellar Contributions to Motor Learning
Authors: Criscimagna-Hemminger Sarah E. , Bastian Amy J. , Shadmehr Reza
Published: February 2010

Small errors may affect the process of learning in a fundamentally different way than large errors. For example, adapting reaching movements in response to a small perturbation produces generalization patterns that are different from large perturbations. Are distinct neural mechanisms engaged in response to large versus small errors? Here, we examined the motor learning process in patients with severe degeneration of the cerebellum. Consistent with earlier reports, we found that the patients were profoundly impaired in adapting their motor commands during reaching movements in response to large, sudden perturbations. However, when the same magnitude perturbation was imposed gradually over many trials, the patients showed marked improvements, uncovering a latent ability to learn from errors. On sudden removal of the perturbation, the patients exhibited aftereffects that persisted much longer than did those in healthy controls. That is, despite cerebellar damage, the brain maintained the ability to learn from small errors and the motor memory that resulted from this learning was strongly resistant to change. Of note was the fact that on completion of learning, the motor output of the cerebellar patients remained distinct from healthy controls in terms of its temporal characteristics. Therefore cerebellar degeneration impaired the ability to learn from large-magnitude errors, but had a lesser impact on learning from small errors. The neural basis of motor learning in response to small and large errors appears to be distinct.
Mapping Human Somatosensory Cortex in Individual Subjects With 7T Functional MRI
Authors: Sanchez-Panchuelo R. M. , Francis S. , Bowtell R. , Schluppeck D.
Published: February 2010

Functional magnetic resonance imaging (fMRI) is now routinely used to map the topographic organization of human visual cortex. Mapping the detailed topography of somatosensory cortex, however, has proven to be more difficult. Here we used the increased blood-oxygen-level-dependent contrast-to-noise ratio at ultra-high field (7 Tesla) to measure the topographic representation of the digits in human somatosensory cortex at 1 mm isotropic resolution in individual subjects. A “traveling wave” paradigm was used to locate regions of cortex responding to periodic tactile stimulation of each distal phalangeal digit. Tactile stimulation was applied sequentially to each digit of the left hand from thumb to little finger (and in the reverse order). In all subjects, we found an orderly map of the digits on the posterior bank of the central sulcus (postcentral gyrus). Additionally, we measured event-related responses to brief stimuli for comparison with the topographic mapping data and related the fMRI responses to anatomical images obtained with an inversion-recovery sequence. Our results have important implications for the study of human somatosensory cortex and underscore the practical utility of ultra-high field functional imaging with 1 mm isotropic resolution for neuroscience experiments. First, topographic mapping of somatosensory cortex can be achieved in 20 min, allowing time for further experiments in the same session. Second, the maps are of sufficiently high resolution to resolve the representations of all five digits and third, the measurements are robust and can be made in an individual subject. These combined advantages will allow somatotopic fMRI to be used to measure the representation of digits in patients undergoing rehabilitation or plastic changes after peripheral nerve damage as well as tracking changes in normal subjects undergoing perceptual learning.
A Discussion Related to Orbit Determination Using Nonlinear SigmaPoint Kalman Filter
Authors: Pardal Paula Cristiane Pinto Mesquita , Kuga Helio Koiti , de Moraes Rodolpho Vilhena
Published: February 2010

Herein, the purpose is to present a Kalman filter based on the sigma point unscented transformation development, aiming at real‐time satellite orbit determination using GPS measurements. First, a brief review of the extended Kalman filter will be done. After, the sigma point Kalman filter will be introduced as well as the basic idea of the unscented transformation, in which this filter is based. Following, the unscented Kalman filter applied to orbit determination will be explained. Such explanation encloses formulations about the orbit determination through GPS; the dynamic model; the observation model; the unmodeled acceleration estimation; also an application of this new filter approaches on orbit determination using GPS measurements discussion.
Biogeography of Tick-Borne Bhanja Virus (Bunyaviridae) in Europe
Authors: Hubálek Zdenek
Published: February 2010

Bhanja virus (BHAV) is pathogenic for young domestic ruminants and also for humans, causing fever and affections of the central nervous system. This generally neglected arbovirus of the familyBunyaviridaeis transmitted by metastriate ticks of the generaHaemaphysalis, Dermacentor, Hyalomma, Rhipicephalus, Boophilus, andAmblyomma. Geographic distribution of BHAV covers southern and Central Asia, Africa, and southern (partially also central) Europe. Comparative biogeographic study of eight known natural foci of BHAV infections in Europe (in Italy, Croatia, Bulgaria, Slovakia) has revealed their common features. (1) submediterranean climatic pattern with dry growing season and wet mild winter (or microlimatically similar conditions, e.g., limestone karst areas in central Europe), (2) xerothermic woodland-grassland ecosystem, with plant alliancesQuercetalia pubescentis, Festucetalia valesiacae, and Brometalia erecti, involving pastoral areas, (3) presence of at least one of the tick speciesHaemaphysalis punctata, Dermacentor marginatus, Rhipicephalus bursa, and/orHyalomma marginatum, and (4) presence of≥60% of the 180 BHAV bioindicator (157 plant, 4 ixodid tick, and 19 vertebrate spp.). On that basis, Greece, France (southern, including Corsica), Albania, Spain, Hungary, European Turkey, Ukraine (southern), Switzerland (southern), Austria (southeastern), Germany (southern), Moldova, and European Russia (southern) have been predicted as additional European regions where BHAV might occur.
Column Strengthening with TRC: Influences of the Column Geometry onto the Confinement Effect
Authors: Ortlepp Regine , Lorenz Andy , Curbach Manfred
Published: February 2010

The increase of the load‐carrying capacity of columns being reinforced with Textile Reinforced Concrete (TRC) is partly achieved by the additional concrete cover. But then it is also decisively caused by the confinement effect of the textile reinforcement. The confinement is thereby producing a three‐axial state of stress within the concrete core of the column. The effectiveness of such a confinement is especially dependent on the geometry of the concrete column to be strengthened. At rectangular ones with sharp edges without ogees the TRC strengthening can only augment the load‐carrying concrete share not create a confinement effect which can be achieved at the round counterparts. Within the study we tested columns with all possible cross‐sections from square to circle with different transition radiuses. Thus the influence of the transition radius onto the local‐bearing capacity of the reinforcing textile was recorded. Furthermore the impact of different fibre materials and reinforcement degrees of the TRC‐strengthening layer has been examined. The first results show a considerable disproportionate increase of the confinement effect with rising transition radius, as well as a growth of the confinement effect with augmenting level of reinforcement in the TRC‐strengthening layer.
A Theoretical Model for Metal Corrosion Degradation
Authors: Svintradze David V. , Pidaparti Ramana M.
Published: February 2010

Many aluminum and stainless steel alloys contain thin oxide layers on the metal surface which greatly reduce the corrosion rate. Pitting corrosion, a result of localized breakdown of such films, results in accelerated dissolution of the underlying metal through pits. Many researchers have studied pitting corrosion for several decades and the exact governing equation for corrosion pit degradation has not been obtained. In this study, the governing equation for corrosion degradation due to pitting corrosion behavior was derived from solid-state physics and some solutions and simulations are presented and discussed.
Convergence of the Sequence of Successive Approximations to a Fixed Point
Authors: Suzuki Tomonari
Published: February 2010

If "Equation missing" is a complete metric space and "Equation missing" is a contraction on "Equation missing", then the conclusion of the Banach-Caccioppoli contraction principle is that the sequence of successive approximations "Equation missing" of "Equation missing" starting from any point "Equation missing" converges to a unique fixed point. In this paper, using the concept of "Equation missing"-distance, we obtain simple, sufficient, and necessary conditions of the above conclusion.
Evaluation of the Proteasome Inhibitor MLN9708 in Preclinical Models of Human Cancer
Authors: Kupperman Erik , Lee Edmund C. , Cao Yueying , Bannerman Bret , Fitzgerald Michael , Berger Allison , Yu Jie , Yang Yu , Hales Paul , Bruzzese Frank , Liu Jane , Blank Jonathan , Garcia Khristofer , Tsu Christopher , Dick Larry , Fleming Paul , Yu Li , Manfredi Mark , Rolfe Mark , Bolen Joe
Published: February 2010

The proteasome was validated as an oncology target following the clinical success of VELCADE (bortezomib) for injection for the treatment of multiple myeloma and recurring mantle cell lymphoma. Consequently, severalgroups are pursuing the development of additional small-molecule proteasome inhibitors for both hematologic and solid tumor indications. Here, we describe MLN9708, a selective, orally bioavailable, second-generation proteasome inhibitor that is in phase I clinical development. MLN9708 has a shorter proteasome dissociation half-life and improved pharmacokinetics, pharmacodynamics, and antitumor activity compared with bortezomib. MLN9708 has a larger blood volume distribution at steady state, and analysis of 20S proteasome inhibition and markers of the unfolded protein response confirmed that MLN9708 has greater pharmacodynamic effects in tissues than bortezomib. MLN9708 showed activity in both solid tumor and hematologic preclinical xenograft models, and we found a correlation between greater pharmacodynamic responses and improved antitumor activity. Moreover, antitumor activity was shown via multiple dosing routes, including oral gavage. Taken together, these data support the clinical development of MLN9708 for both hematologic and solid tumor indications. Cancer Res; 70(5); 1970–80
Talin1 Promotes Tumor Invasion and Metastasis via Focal Adhesion Signaling and Anoikis Resistance
Authors: Sakamoto Shinichi , McCann Richard O. , Dhir Rajiv , Kyprianou Natasha
Published: February 2010

Talin1 is a focal adhesion complex protein that regulates integrin interactions with ECM. This study investigated the significance of talin1 in prostate cancer progression to metastasis in vitro and in vivo. Talin1 overexpression enhanced prostate cancer cell adhesion, migration, and invasion by activating survival signals and conferring resistance to anoikis. ShRNA-mediated talin1 loss led to a significant suppression of prostate cancer cell migration and transendothelial invasion in vitro and a significant inhibition of prostate cancer metastasis in vivo. Talin1-regulated cell survival signals via phosphorylation of focal adhesion complex proteins, such as focal adhesion kinase and Src, and downstream activation of AKT. Targeting AKT activation led to a significant reduction of talin1-mediated prostate cancer cell invasion. Furthermore, talin1 immunoreactivity directly correlated with prostate tumor progression to metastasis in the transgenic adenocarcinoma mouse prostate mouse model. Talin1 profiling in human prostate specimens revealed a significantly higher expression of cytoplasmic talin1 in metastatic tissue compared with primary prostate tumors (P < 0.0001). These findings suggest (a) a therapeutic significance of disrupting talin1 signaling/focal adhesion interactions in targeting metastatic prostate cancer and (b) a potential value for talin1 as a marker of tumor progression to metastasis. Cancer Res; 70(5); 1885–95
A Bax-Mediated Mechanism for Obatoclax-Induced Apoptosis of Cholangiocarcinoma Cells
Authors: Smoot Rory L. , Blechacz Boris R.A. , Werneburg Nathan W. , Bronk Steve F. , Sinicrope Frank A. , Sirica Alphonse E. , Gores Gregory J.
Published: February 2010

Apoptosis induction by BH3 mimetics is a therapeutic strategy for human cancer. These mimetics exert single-agent activity in cells “primed” for cell death. Primed cells are dependent upon antiapoptotic Bcl-2 proteins for survival and are characterized by the ability of the BH3 mimetic to induce cytochrome c release from their isolated mitochondria. Our aim was to examine the single-agent activity of obatoclax, a BH3 mimetic in cholangiocarcinoma cell lines. In clonogenic assays, inhibition of colony formation was observed by obatoclax treatment. Despite single-agent activity by obatoclax, the mitochondria from these cells did not release cytochrome c after incubation with this BH3 mimetic. However, immunofluorescence and cell fractionation studies identified Bax activation and translocation to mitochondria after treatment with obatoclax. shRNA targeted knockdown of Bax doubled the IC50 for obatoclax but did not abrogate its cytotoxicity, whereas knockdown of Bak did not alter the IC50. In a cell-free system, obatoclax induced an activating conformational change of Bax, which was attenuated by a site-directed mutagenesis of a previously identified protein activation site. Finally, the drug also elicited a significant in vivo response in a rodent model of this disease. In conclusion, single-agent obatoclax treatment results in Bax activation, which contributes, in part, to cell death in cholangiocarcinoma cells. These data indicate that BH3 mimetics may also function as direct activators of Bax and induce cytotoxicity in cells not otherwise primed for cell death. Cancer Res; 70(5); 1960–9
The COXEN Principle: Translating Signatures of In vitro Chemosensitivity into Tools for Clinical Outcome Prediction and Drug Discovery in Cancer
Authors: Smith Steven C. , Baras Alexander S. , Lee Jae K. , Theodorescu Dan
Published: February 2010

Substantial effort has been devoted to in vitro testing of candidate chemotherapeutic agents. In particular, the United States National Cancer Institute Developmental Therapeutics Program (NCI-DTP) Human Tumor Cell Line Screen has screened hundreds of thousands of compounds and extracts, for which data on more than 40,000 compounds tested on a panel of 60 cancer cell lines (NCI-60) are publically available. In tandem, gene expression profiling has brought about a sea change in our understanding of cancer biology, allowing discovery of biomarkers or signatures able to characterize, classify, and prognosticate clinical behavior of human tumors. Recent studies have used tumor profiling matched to clinical trial outcome data to derive gene expression models predicting therapeutic outcomes, though such efforts are costly, time-consuming, tumor type-specific, and not amenable to rare diseases. Furthermore, addition of new or established drugs to multidrug combinations in which such models are already available requires the entire model to be re-derived. Can the aforementioned in vitro testing platform, coupled to the universal language of genomics, be used to develop, a priori, gene expression models predictive of clinical outcomes? Recent advances, including the CO-eXpression ExtrapolatioN (COXEN) algorithm, suggest that development of these models may be possible and raise important implications for future trial design and drug discovery. Cancer Res; 70(5); 1753–8
Protein Kinase D1 Inhibits Cell Proliferation through Matrix Metalloproteinase-2 and Matrix Metalloproteinase-9 Secretion in Prostate Cancer
Authors: Biswas M. Helal Uddin , Du Cheng , Zhang Chuanyou , Straubhaar Juerg , Languino Lucia R. , Balaji K.C.
Published: February 2010

We and others previously showed that protein kinase D1 (PKD1) is downregulated in several cancers including prostate; interacts with E-cadherin, a major cell adhesion epithelial protein; and causes increased cell aggregation and decreased motility of prostate cancer cells. In this study, we show that PKD1 complexes with β3-integrin, resulting in activation of mitogen-activated protein kinase/extracellular signal-regulated kinase (ERK) kinase–ERK pathway, which causes increased production of matrix metalloproteinase (MMP)-2 and MMP-9, that is associated with shedding of soluble 80 kDa E-cadherin extracellular domain. Interestingly, decreased cell proliferation following PKD1 transfection was rescued by MMP-2 and MMP-9 inhibitors and augmented by recombinant MMP-2 (rMMP-2) and rMMP-9 proteins, suggesting an antiproliferative role for MMPs in prostate cancer. Translational studies by in silico analysis of publicly available DNA microarray data sets show a significant direct correlation between PKD1 and MMP-2 expression in human prostate tissues. The study shows a novel mechanism for antiproliferative effects of PKD1, a protein of emerging translational interest in several human cancers, through increased production of MMP-2 and MMP-9 in cancer cells. Cancer Res; 70(5); 2095–104
ZIP4 Regulates Pancreatic Cancer Cell Growth by Activating IL-6/STAT3 Pathway through Zinc Finger Transcription Factor CREB
Authors: Zhang Yuqing , Bharadwaj Uddalak , Logsdon Craig D. , Chen Changyi , Yao Qizhi , Li Min
Published: February 2010

Purpose: Recent studies indicate a strong correlation of zinc transporter ZIP4 and pancreatic cancer progression; however, the underlying mechanisms are unclear. We have recently found that ZIP4 is overexpressed in pancreatic cancer. In this study, we investigated the signaling pathway through which ZIP4 regulates pancreatic cancer growth.

Experimental Design: The expression of cyclin D1, interleukin 6 (IL-6), and signal transducer and activator of transcription 3 (STAT3) in pancreatic cancer xenografts and cells were examined by real-time PCR, Bio-Plex cytokine assay, and Western blot, respectively. The activity of cAMP response element–binding protein (CREB) is examined by a promoter activity assay.

Results: Cyclin D1 was significantly increased in the ZIP4 overexpressing MIA PaCa-2 cells (MIA-ZIP4)–injected orthotopic xenografts and was downregulated in the ZIP4-silenced ASPC-1 (ASPC-shZIP4) group. The phosphorylation of STAT3, an upstream activator of cyclin D1, was increased in MIA-ZIP4 cells and decreased in ASPC-shZIP4 cells. IL-6, a known upstream activator for STAT3, was also found to be significantly increased in the MIA-ZIP4 cells and xenografts and decreased in the ASPC-shZIP4 group. Overexpression of ZIP4 led to a 75% increase of IL-6 promoter activity and caused increased phosphorylation of CREB.

Conclusions: Our study suggest that ZIP4 overexpression causes increased IL-6 transcription through CREB, which in turn activates STAT3 and leads to increased cyclin D1 expression, resulting in increased cell proliferation and tumor progression in pancreatic cancer. These results elucidated a novel pathway in ZIP4-mediated pancreatic cancer growth and suggest new therapeutic targets, including ZIP4, IL-6, and STAT3, in pancreatic cancer treatment. Clin Cancer Res; 16(5); 1423–30
Diverse Patterns of T-Cell Response against Multiple Newly Identified Human Y Chromosome–Encoded Minor Histocompatibility Epitopes
Authors: Ofran Yishai , Kim Haesook T. , Brusic Vladimir , Blake Loren , Mandrell Michael , Wu Catherine J. , Sarantopoulos Stefanie , Bellucci Roberto , Keskin Derin B. , Soiffer Robert J. , Antin Joseph H. , Ritz Jerome
Published: February 2010

Purpose: Donor T cells respond to minor histocompatibility antigens (mHA), resulting in both graft-versus-host disease and graft versus leukemia after allogeneic hematopoietic stem cell transplantation. Because relatively few mHAs are known, we developed a new approach to predict and subsequently validate candidate mHA.

Experimental Design: We developed an algorithm based on genetic disparities between Y chromosome–encoded and X chromosome–encoded proteins and known requirements for binding to HLA class I molecules to predict Y chromosome–derived, HLA A*0201–restricted peptides (HY) and ranked peptides based on potential immunogenicity. We evaluated T-cell responses to 41 candidate peptides in 28 male recipients with female donors (FM), 22 male recipients with male donors (MM), and 26 normal individuals. All patients and donors were HLA A*0201 positive.

Results: Thirteen peptides derived from five proteins elicited significantly greater T-cell responses in FM patients compared with MM patients and in normal females compared with normal males. Six peptides were more immunogenic than the only previously known HLA A*0201–restricted Y-encoded mHA. Twenty-seven of 28 FM patients responded to at least one HY peptide, but despite a common Y chromosome mismatch and expression of HLA A*0201, each patient responded to a unique set of peptides.

Conclusions: Novel HLA A*0201–restricted HY epitopes can be predicted and validated in patients after allogeneic hematopoietic stem cell transplantation. Highly diverse patterns of T-cell response against these epitopes have been identified. Prospective monitoring of responses to large panels of immunogenic peptides can facilitate the identification of clinically relevant targets of graft-versus-host disease and graft versus leukemia. Clin Cancer Res; 16(5); 1642–51
Association of Polymorphisms inAKT1andEGFRwith Clinical Outcome and Toxicity in Non–Small Cell Lung Cancer Patients Treated with Gefitinib
Authors: Giovannetti Elisa , Zucali Paolo A. , Peters Godefridus J. , Cortesi Filippo , D'Incecco Armida , Smit Egbert F. , Falcone Alfredo , Burgers Jacobus A. , Santoro Armando , Danesi Romano , Giaccone Giuseppe , Tibaldi Carmelo
Published: February 2010

EGFR mutations are strongly predictive of epidermal growth factor receptor (EGFR)-tyrosine kinase inhibitor activity in non–small cell lung cancer (NSCLC), but resistance mechanisms are not completely understood. The interindividual variability in toxicity also points out to the need of novel pharmacogenetic markers to select patients before therapy. Therefore, we evaluated the associations between EGFR and AKT1 polymorphisms and outcome/toxicity in gefitinib-treated NSCLC patients. Polymorphic loci in EGFR, and AKT1, and EGFR and K-Ras mutations were assessed in DNA isolated from blood samples and/or paraffin-embedded tumor from 96 gefitinib-treated NSCLC patients. Univariate and multivariate analyses compared genetic variants with clinical efficacy and toxicity using Fisher's, log-rank test, and Cox's proportional hazards model. AKT1-SNP4 association with survival was also evaluated in 127 chemotherapy-treated/gefitinib-naive patients, whereas its relationship with AKT1 expression and gefitinib cytotoxicity was studied in 15 NSCLC cell lines. AKT1-SNP4 A/A genotype was associated with shorter time-to-progression (P = 0.04) and overall survival (P = 0.007). Multivariate analyses and comparison with the gefitinib-nontreated population underlined its predictive significance, whereas the in vitro studies showed the association of lower AKT1 mRNA levels with gefitinib resistance. In contrast, EGFR-activating mutations were significantly correlated with response, longer time-to-progression, and overall survival, whereas EGFR −191C/A (P < 0.001), −216 G/T (P < 0.01), and R497K (P = 0.02) polymorphisms were strongly associated with grade >1 diarrhea. AKT1-SNP4 A/A genotype seems to be a candidate biomarker of primary resistance, whereas EGFR −191C/A, −216G/T, and R497K polymorphisms are associated with diarrhea when using gefitinib in NSCLC patients, thus offering potential new tools for treatment optimization. Mol Cancer Ther; 9(3); 581–93
Reduced Argininosuccinate Synthetase Is a Predictive Biomarker for the Development of Pulmonary Metastasis in Patients with Osteosarcoma
Authors: Kobayashi Eisuke , Masuda Mari , Nakayama Robert , Ichikawa Hitoshi , Satow Reiko , Shitashige Miki , Honda Kazufumi , Yamaguchi Umio , Shoji Ayako , Tochigi Naobumi , Morioka Hideo , Toyama Yoshiaki , Hirohashi Setsuo , Kawai Akira , Yamada Tesshi
Published: February 2010

Pulmonary metastasis is the most significant prognostic determinant for osteosarcoma, but methods for its prediction and treatment have not been established. Using oligonucleotide microarrays, we compared the global gene expression of biopsy samples between seven osteosarcoma patients who developed pulmonary metastasis within 4 years after neoadjuvant chemotherapy and curative resection, and 12 patients who did not relapse. We identified argininosuccinate synthetase (ASS) as a gene differentially expressed with the highest statistical significance (Welch's t test, P = 2.2 × 10−5). Immunohistochemical analysis of an independent cohort of 62 osteosarcoma cases confirmed that reduced expression of ASS protein was significantly correlated with the development of pulmonary metastasis after surgery (log-rank test, P < 0.05). Cox regression analysis revealed that ASS was the sole significant predictive factor (P = 0.039; hazard ratio, 0.319; 95% confidence interval, 0.108-0.945). ASS is one of the enzymes required for the production of a nonessential amino acid, arginine. We showed that osteosarcoma cells lacking ASS expression were auxotrophic for arginine and underwent G0-G1 arrest in arginine-free medium, suggesting that an arginine deprivation therapy could be effective in patients with osteosarcoma. Recently, phase I and II clinical trials in patients with melanoma and hepatocellular carcinoma have shown the safety and efficacy of plasma arginine depletion by stabilized arginine deiminase. Our data indicate that in patients with osteosarcoma, reduced expression of ASS is not only a novel predictive biomarker for the development of metastasis, but also a potential target for pharmacologic intervention. Mol Cancer Ther; 9(3); 535–44
Epidermal Growth Factor–Induced Heparanase Nucleolar Localization Augments DNA Topoisomerase I Activity in Brain Metastatic Breast Cancer
Authors: Zhang Lixin , Sullivan Peggy , Suyama Julie , Marchetti Dario
Published: February 2010

Identification of molecular mechanisms responsible for brain metastatic breast cancer (BMBC) is imperative to develop novel therapies. However, current understanding of the molecular circuitry that governs BMBC dissemination remains fragmentary. Heparanase (HPSE) is the only functional mammalian endoglycosidase whose activity correlates with cancer metastasis, angiogenesis, and the reduced postoperative survival of cancer patients, making it an active target for anticancer therapeutics. We hypothesized that human epidermal growth factor receptor 2 (HER2)/epidermal growth factor receptor (EGFR) activation promotes HPSE function in human BMBC. To address this, we examined HPSE content, activity, and intracellular trafficking in a HER2/EGFR-expressing BMBC model system and show that HPSE is present, functional, and correlates with HER2 status. Further, we showed that EGF induced nucleolar translocation of HPSE in these cells in a dose- and time-dependent manner upon activation of HER2/EGFR. Knockdowns of HER2/EGFR by small interference RNA abolished EGF-induced HPSE nucleolar translocalization. It was also noted that nucleolar HPSE modulates DNA topoisomerase I (Topo I), an enzyme that is highly present in nucleoli, essential for DNA replication and transcription in a variety of tumors, and inhibited by heparan sulfate. Evidence is provided that HPSE can regulate Topo I activity, which subsequently affects BMBC cell proliferation. Finally, we showed that the nucleolar presence of HPSE with Topo I colocalization is detected only in HER2-overexpressing BMBC patient specimens. Altogether, these findings support the notion that HPSE is a critical downstream target of HER2 mechanisms driving BMBC and is potentially relevant for BMBC therapeutic interventions. Mol Cancer Res; 8(2); 278–90
Preimplantation Genetic Diagnosis (Embryo Screening) for Enlarged Vestibular Aqueduct due to SLC26A4 Mutation
Authors: Wu Chen-Chi , Lin Shin-Yu , Su Yi-Nin , Fang Mei-Ya , Chen Shee-Uan , Hsu Chuan-Jen
Published: February 2010

Preimplantation genetic diagnosis (PGD) is used to analyze embryos genetically before their transfer into the uterus. For families with genetic diseases, PGD offers a chance to have an unaffected child, without facing termination of pregnancy. Although PGD has been performed for many monogenic disorders, such as cystic fibrosis and β-thalassemia, the application of PGD to hereditary hearing impairment has not been explored. In the present study, we reported the development and application of PGD protocols to address enlarged vestibular aqueduct (EVA), which is a common type of hereditary hearing impairment associated with mutations in the SLC26A4 gene. The family requesting PGD had a history of EVA, segregating the SLC26A4 c.919-2A→G mutation. In short, the PGD process was composed of two steps: the development of a single-cell testing protocol and clinical PGD cycles (i.e., selection and implantation of unaffected embryos using the single-cell testing protocol). First, protocols for genetic testing in a single cell were established for the c.919-2A→G mutation using GenomiPhi technology and primer extension mini-sequencing. These protocols were validated on single lymphocytes collected from both parents and their affected child. Two clinical PGD cycles were then performed for the parents, with the second cycle successfully leading to a singleton pregnancy. The baby was homozygous for the wild-type SLC26A4 allele and revealed a normal audiological phenotype after birth. To our knowledge, this is the first report in the literature describing successful PGD in families with genetic hearing impairment. In our opinion, the application of PGD in the field of hereditary hearing impairment involves fewer ethical controversies than other novel applications of PGD and traditional indications for PGD for other monogenic diseases. Therefore, the approach demonstrated in the present study can also be used in a large number of families with other types of hereditary hearing impairment.
Mesangial Cells Initiate Compensatory Tubular Cell Hypertrophy
Authors: Sinuani I. , Beberashvili I. , Averbukh Z. , Cohn M. , Gitelman I. , Weissgarten J.
Published: February 2010

Unilateral nephrectomy results in compensatory renal growth, in which both the size and the functional capacity of the remaining kidney are increased. The functional adaptation to the removal of the contralateral kidney consists mostly of an increase in the glomerular filtration rate of the remaining kidney, and hypertrophy of cells comprising the nephron, mainly of the proximal tubular cells. Although the phenomenon of single kidney hypertrophy has been known for the past thousand years and despite intensive research over the past century, the mechanism of this process still remains unclear. The present article reviews the role of mesangial cells in compensatory renal hypertrophy.
Prospective Controlled Animal Study on Biopsy Sampling with New Flexible Cryoprobes versus Forceps: Evaluation of Biopsy Size, Histological Quality and Bleeding Risk
Authors: Franke Karl-Josef , Theegarten Dirk , Hann von Weyhern Claus , Nilius Georg , Brueckner Christiane , Hetzel Jürgen , Hetzel Martin , Ruhle Karl-Heinz , Enderle Markus D. , Szyrach Mara N.I.
Published: February 2010

Background: Cryoextraction is a procedure used for the recanalization of obstructed airways caused by visible exophytic endobronchial tumor. Biopsy samples obtained by this technique have been shown to be useful for histological assessment. Objectives: The aim of the present animal study was to systematically evaluate biopsy size, histological quality and bleeding risk after cryobiopsy with new, flexible cryoprobes in comparison with forceps biopsy, serving as the gold standard. Methods: Biopsies were obtained from anesthetized pigs with the flexible bronchoscopy technique, and evaluated histologically with respect to their size and quality. Bleeding frequency, bleeding duration and histological changes in the biopsy bed were also recorded. Results: Cryobiopsies were significantly larger than forceps biopsies. The size of cryobiopsies was dependent on the freezing time. The histological quality of the cryobiopsy specimenswas not impaired by the freezing process, whereas forceps biopsies showed typical crush artifacts. Despite the larger defects left in the tracheobronchial system after cryobiopsy, bleeding frequency and duration were not higher compared to forceps biopsy. Conclusions: Since cryobiopsy sampling is not associated with a higher bleeding risk compared with forceps biopsy, this new biopsy technique offers – in addition to a good specimen quality – a safe and valuable tool with the potential of improving the outcome of diagnostic endoscopy.
Evaluation of the American Heart Association Cardiovascular Disease Prevention Guideline for Women
Authors: Hsia Judith , Rodabough Rebecca J. , Manson JoAnn E. , Liu Simin , Freiberg Matthew S. , Graettinger William , Rosal Milagros C. , Cochrane Barb , Lloyd-Jones Donald , Robinson Jennifer G. , Howard Barbara V.
Published: February 2010

Background—

The 2007 update to the American Heart Association (AHA) guidelines for cardiovascular disease prevention in women recommend a simplified approach to risk stratification. We assigned Women’s Health Initiative participants to risk categories as described in the guideline and evaluated clinical event rates within and between strata.

Methods and Results—

The Women’s Health Initiative enrolled 161 808 women ages 50 to 79 years and followed them prospectively for 7.8 years (mean). Applying the 2007 AHA guideline categories, 11% of women were high risk, 72% at-risk, and 4% at optimal risk; 13% of women did not fall into any category, that is, lacked risk factors but did not adhere to a healthy lifestyle (moderate intensity exercise for 30 minute most days and <7% of calories from saturated fat). Among high risk, at-risk, and optimal risk women, rates of myocardial infarction/coronary death were 12.5%, 3.1%, and 1.1% per 10 years (
P
for trend <0.0001); the event rate was 1.3% among women who could not be categorized. We observed a graded relationship between risk category and cardiovascular event rates for white, black, Hispanic, and Asian women, although event rates differed among ethnic groups (
P
for interaction=0.002). The AHA guideline predicted coronary events with accuracy similar to current Framingham risk categories (area under receiver operating characteristic curve for Framingham risk, 0.665; for AHA risk, 0.664;
P
=0.94) but less well than proposed Framingham 10-year risk categories of <5%, 5% to 20%, and >20% (area under receiver operating characteristic curve for Framingham risk, 0.724; for AHA risk, 0.664;
P
<0.0001).

Conclusions—

Risk stratification as proposed in the 2007 AHA guideline is simple, accessible to patients and providers, and identifies cardiovascular risk with accuracy similar to that of the current Framingham algorithm.

Clinical Trial Registration—

clinicaltrials.gov. Identifier: NCT00000611.
Ubiquitin Proteasome Dysfunction in Human Hypertrophic and Dilated Cardiomyopathies
Authors: Predmore Jaime M. , Wang Ping , Davis Frank , Bartolone Sarah , Westfall Margaret V. , Dyke David B. , Pagani Francis , Powell Saul R. , Day Sharlene M.
Published: February 2010

Background—

The ubiquitin proteasome system maintains a dynamic equilibrium of proteins and prevents accumulation of damaged and misfolded proteins, yet its role in human cardiac dysfunction is not well understood. The present study evaluated ubiquitin proteasome system function in human heart failure and hypertrophic cardiomyopathy (HCM).

Methods and Results—

Proteasome function was studied in human nonfailing donor hearts, explanted failing hearts, and myectomy samples from patients with HCM. Proteasome proteolytic activities were markedly reduced in failing and HCM hearts compared with nonfailing hearts (
P
<0.01). This activity was partially restored after mechanical unloading in failing hearts (
P
<0.01) and was significantly lower in HCM hearts with pathogenic sarcomere mutations than in those lacking these mutations (
P
<0.05). There were no changes in the protein content of ubiquitin proteasome system subunits (ie, 11S, 20S, and 19S) or in active-site labeling of the 20S proteolytic subunit β-5 among groups to explain decreased ubiquitin proteasome system activity in HCM and failing hearts. Examination of protein oxidation revealed that total protein carbonyls, 4-hydroxynonenylated proteins, and oxidative modification to 19S ATPase subunit Rpt 5 were increased in failing compared with nonfailing hearts.

Conclusions—

Proteasome activity in HCM and failing human hearts is impaired in the absence of changes in proteasome protein content or availability of proteolytic active sites. These data provide strong evidence that posttranslational modifications to the proteasome may account for defective protein degradation in human cardiomyopathies.
Impacts of Assimilating AMMA Soundings on ECMWF Analyses and Forecasts
Authors: Agustí-Panareda Anna , Beljaars Anton , Cardinali Carla , Genkova Iliana , Thorncroft Chris
Published: February 2010

The field experiment of the African Monsoon Multidisciplinary Analysis (AMMA) project during the 2006 wet monsoon season provided an unprecedented amount of radiosonde/dropsonde data over the West African region. This paper explores the usage and impacts of this invaluable dataset in the European Centre for Medium-Range Weather Forecasts analyses and forecasts. These soundings are the only source of data that can provide 3D information on the thermodynamic and dynamic structures of the lower troposphere over continental West Africa. They are particularly important for the Sahel region located between 12° and 20°N, which is characterized by large gradients in temperature and moisture in the lower troposphere. An assimilation experiment comparison between the pre-AMMA and AMMA radiosonde networks shows that the extra AMMA soundings have a significant analysis impact on the low-level temperature over the Sahel and on the structure of the African easterly jet. However, the impacts of the extra AMMA data on the forecast disappear after 24 h. The soundings reveal large model biases in boundary layer temperature over the northern and eastern Sahel, which are consistent with the well-known model biases in cloud, rainfall, and radiation. Large analysis increments in temperature lead to increments in divergence and subsidence, which act to suppress convection. Thus, the analysis increments appear to have an undesirable feedback on the cloud and temperature model biases. The impact of the AMMA soundings on the African easterly jet is to enhance and extend the jet streak to 15°E, that is, toward the eastern part of the Sahel. No observations are assimilated east of 15°E at the level of the African easterly jet to support the jet enhancement farther east. Comparisons with independent atmospheric cloud motion vectors indicate that the African easterly jet in the analysis is too weak over this data-sparse region. This could have implications for the development of African easterly waves in the model forecast. Further experimentation by assimilating atmospheric motion vectors—currently not used—could address this problem.