bluuebunny/crossref_metadata_2025 huggingface dataset:
Intraoperative qRT-PCR for Detection of Lymph Node Metastasis in Head and Neck Cancer
Authors: Ferris Robert L. , Xi Liqiang , Seethala Raja R. , Chan Jon , Desai Shaun , Hoch Benjamin , Gooding William , Godfrey Tony E.
Published: February 2011

Purpose: Sentinel node biopsy (SNB) has been shown to accurately stage the regional lymphatics in oral carcinoma. However, intraoperative pathology is only moderately sensitive and final pathology takes several days to complete. The purpose of this study was to develop a rapid, automated, and quantitative real-time PCR (qRT-PCR) assay that can match final pathology in an intraoperative time frame.

Experimental Design: Four hundred forty-eight grossly tumor-negative lymph nodes were evaluated for expression of 3 markers [PVA (pemphigus vulgaris antigen), PTHrP (parathyroid hormone-related protein), and TACSTD1 (tumor-associated calcium signal transducer 1)]. Conformity of metastasis detection by qRT-PCR was determined using hematoxylin and eosin and immunohistochemistry staining as the gold standard. PVA and TACSTD1 were then multiplexed with β-glucuronidase to develop a rapid, automated single-tube qRT-PCR assay using the Cepheid GeneXpert system. This assay was used to analyze 103 lymph nodes in an intraoperative time frame.

Results: Four hundred forty-two nodes produced an informative result for both qRT-PCR and pathologic examination. Concordance of qRT-PCR for individual markers with final pathology ranged from 93% to 98%. The best marker combination was TACSTD1 and PVA. A rapid, multiplex assay for TACSTD1 and PVA was developed on the Cepheid GeneXpert and demonstrated an excellent reproducibility and linearity. Analysis of 103 lymph nodes demonstrated 94.2% accuracy of this assay for identifying positive and negative nodes. The average time for each assay to yield results was 35 minutes.

Conclusions: A rapid, automated qRT-PCR assay can detect lymph node metastasis in head and neck cancer with high accuracy compared to pathologic analysis and may be more accurate than intraoperative pathology. Combined, SNB and rapid qRT-PCR could more appropriately guide surgical treatment of patients with head and neck cancer. Clin Cancer Res; 17(7); 1858–66. ©2011 AACR.
Peptide-Loaded Langerhans Cells, Despite Increased IL15 Secretion and T-Cell ActivationIn Vitro, Elicit Antitumor T-Cell Responses Comparable to Peptide-Loaded Monocyte-Derived Dendritic CellsIn Vivo
Authors: Romano Emanuela , Rossi Marco , Ratzinger Gudrun , de Cos Maria-Angeles , Chung David J. , Panageas Katherine S. , Wolchock Jedd D. , Houghton Alan N. , Chapman Paul B. , Heller Glenn , Yuan Jianda , Young James W.
Published: February 2011

Purpose: We compared the efficacy of human Langerhans cells (LC) as tumor immunogens in vivo with monocyte-derived dendritic cells (moDC) and investigated how interleukin 15 (IL15) supports optimal DC-stimulated antitumor immunity.

Experimental Design: American Joint Committee on Cancer stage III/IV melanoma patients participated in this first clinical trial comparing melanoma peptide-pulsed LC with moDC vaccines (NCT00700167, www.ClinicalTrials.gov). Correlative studies evaluated mechanisms mediating IL15 support of DC-stimulated antitumor immunity.

Results: Both DC vaccines were safe and immunogenic for melanoma antigens. LC-based vaccines stimulated significantly greater tyrosinase–HLA-A*0201 tetramer reactivity than the moDC-based vaccines. The two DC subtypes were otherwise statistically comparable, in contrast to extensive prior data in vitro showing LC superiority. LCs synthesize much more IL15 than moDCs and stimulate significantly more antigen-specific lymphocytes with a cytolytic IFN-γ profile even without exogenous IL15. When supplemented by low-dose IL15, instead of IL2, moDCs stimulate 5 to 6 logs more tumor antigen–specific effector memory T cells (TEMRA) over 3 to 4 weeks in vitro. IL2 and IL15 can be synergistic in moDC stimulation of cytolytic T cells. IL15 promotes T-cell expression of the antiapoptotic bcl-2 and inhibits candidate regulatory T-cell (Treg) expansion after DC stimulation, countering two effects of IL2 that do not foster tumor immunity.

Conclusions: MoDC-based vaccines will require exogenous IL15 to achieve clinical efficacy. Alternatively, LCs can couple the endogenous production of IL15 with potent T-cell stimulatory activity. Optimization of full-length tumor antigen expression for processing into multiple immunogenic peptides for presentation by both class I and II MHC therefore merits emphasis to support more effective antitumor immunity stimulated by LCs. Clin Cancer Res; 17(7); 1984–97. ©2011 AACR.
Identification of CD44 as a Surface Biomarker for Drug Resistance by Surface Proteome Signature Technology
Authors: Cain Jason W. , Hauptschein Robert S. , Stewart Jean K. , Bagci Tugba , Sahagian Gary G. , Jay Daniel G.
Published: February 2011

We developed surface proteome signatures (SPS) for identification of new biomarkers playing a role in cancer drug resistance. SPS compares surface antigen expression of different cell lines by immunocytochemistry of a phage display antibody library directed to surface antigens of HT1080 fibrosarcoma cells. We applied SPS to compare the surface proteomes of two epithelial derived cancer cell lines, MCF7 and NCI/ADR-RES, which is drug resistant because of overexpression of the P-glycoprotein (P-gp) drug efflux pump. Surface proteomic profiling identified CD44 as an additional biomarker that distinguishes between these two cell lines. CD44 immunohistochemistry can distinguish between tumors derived from these lines and predict tumor response to doxorubicin in vivo. We further show that CD44 plays a role in drug resistance, independently of P-gp, in NCI/ADR-RES cells and increases expression of the antiapoptotic protein Bcl-xL. Our findings illustrate the utility of SPS to distinguish between cancer cell lines and their derived tumors and identify novel biomarkers involved in drug resistance. Mol Cancer Res; 9(5); 637–47. ©2011 AACR.
Mechanisms of Drug Sensitization to TRA-8, an Agonistic Death Receptor 5 Antibody, Involve Modulation of the Intrinsic Apoptotic Pathway in Human Breast Cancer Cells
Authors: Amm Hope M. , Zhou Tong , Steg Adam D. , Kuo Huichien , Li Yufeng , Buchsbaum Donald J.
Published: February 2011

TRA-8, a monoclonal antibody to death receptor 5 induces apoptosis in various cancer cells; however, the degree of sensitivity varies from highly sensitive to resistant. We have previously shown that resistance to TRA-8 can be reversed by using chemotherapeutic agents, but the mechanism underlying this sensitization was not fully understood. Here, we examined the combination of TRA-8 with doxorubicin or bortezomib in breast cancer cells. In TRA-8–resistant BT-474 and T47D cells, both chemotherapy agents synergistically sensitized cells to TRA-8 cytotoxicity with enhanced activation of apoptosis shown by cleavage of caspases and PARP, reduced Bid, increased proapoptotic Bcl-2 proteins, and increased mitochondrial membrane depolarization. Doxorubicin or bortezomib combined with TRA-8 also reduced Bcl-XL and X-linked inhibitors of apoptosis (XIAP) in treated cells. Furthermore, targeting these proteins with pharmacologic modulators, AT-101, BH3I-2′ and AT-406, produced sensitization to TRA-8. TRA-8 combined with AT-101 or BH3I-2′, inhibitors of antiapoptotic Bcl-2 proteins, produced synergistic cytotoxicity against ZR-75-1, BT-474, and T47D cells. The IAP-targeting compound, AT-406, was synergistic with TRA-8 in BT-474 cells, and to a lesser extent T47D cells. Activation of the intrinsic apoptotic pathway was a common mechanism associated with sensitization of TRA-8–resistant breast cancer cell lines. Collectively, these studies show that the Bcl-2 and IAP families of proteins are involved in TRA-8 and chemotherapy resistance via their modulation of the intrinsic apoptotic pathway. Targeting these proteins with novel agents sensitized TRA-8–resistant breast cancer cells, suggesting this approach may represent a potent therapeutic strategy in the treatment of breast cancer. Mol Cancer Res; 9(4); 403–17. ©2011 AACR.
Upregulation of ABCG2 by Romidepsin via the Aryl Hydrocarbon Receptor Pathway
Authors: To Kenneth K.W. , Robey Robert , Zhan Zhirong , Bangiolo Lois , Bates Susan E.
Published: February 2011

Histone deacetylase inhibitors (HDACI) are promising anticancer agents and their use in combination with conventional anticancer drugs is currently under investigation. We previously reported cell line–specific upregulation of ABCG2, a multidrug resistance transporter shown to control oral bioavailability and CNS penetration, by the HDACI romidepsin, although the precise mechanism in a particular cell line remains to be determined. The aryl hydrocarbon receptor (AhR) is a ligand-dependent transcription factor that can be activated by numerous environmental contaminants and has been shown to be a client protein of heat shock protein 90 (Hsp90). A xenobiotic response element was defined in the ABCG2 promoter and was shown to mediate AhR signaling. Activated AhR was found to be associated with the ABCG2 promoter only in cell line models that respond to romidepsin with ABCG2 upregulation. Our data suggest that romidepsin acetylated Hsp70 and inhibited the chaperone function of Hsp90, thereby allowing the dissociation of AhR from Hsp90. The dissociation of AhR from Hsp90 may be a prerequisite for the differential upregulation of ABCG2 by romidepsin. Increasing our understanding of the mechanism(s) governing differential upregulation of ABCG2 in response to romidepsin could provide an insight into strategies needed to tackle resistance to HDACIs in cancer therapeutics. Mol Cancer Res; 9(4); 516–27. ©2011 AACR.
Activation of Interferon-Stimulated Genes by γ-Ray Irradiation Independently of the Ataxia Telangiectasia Mutated–p53 Pathway
Authors: Sugihara Takashi , Murano Hayato , Nakamura Masako , Ichinohe Kazuaki , Tanaka Kimio
Published: February 2011

The ataxia telangiectasia mutated (ATM)-p53 pathway is a well-known main signal transduction pathway for cellular responses, which is activated by γ-ray irradiation. Microarray analysis showed changes in the expressions of IFN-stimulated genes (ISG) in γ-ray–irradiated Balb/cA/Atm-deficient mouse embryonic fibroblasts (MEF) (ATM-KO), indicating that another pathway for cellular responses besides the ATM-p53 pathway was activated by γ-ray irradiation. The basal expression levels of Irf7 and Stat1 in ATM-KO and p53-deficient MEFs (p53-KO) were higher than those in Atm–wild-type MEFs (ATM-WT) and p53-wild-type MEFs (p53-WT), respectively. Irradiation stimulated the expressions of Irf7 and Stat1 in ATM-KO, p53-KO, ATM-WT, and p53-WT, indicating that upregulation of Irf7 and Stat1 expressions by irradiation did not depend on the ATM-p53 pathway. When conditioned medium (CM) obtained from irradiated ATM-WT or ATM-KO was added to nonirradiated MEFs, the expressions of Irf7 and Stat1 increased. We predicted that gene activation in nonirradiated MEFs was caused by IFN-α/β. Unexpectedly, significant amount of IFN-α/β could not be detected in the CM from irradiated ATM-WT or ATM-KO. Meanwhile, increased expression of Ccl5 (RANTES) protein was detected in the CM from irradiated MEFs. These results indicate that ISGs were activated by γ-ray irradiation independently of the ATM-p53 pathway and gene activation was caused by radiation-induced soluble factors. Mol Cancer Res; 9(4); 476–84. ©2011 AACR.
The Adverse Impact of Diabetes Mellitus on Left Ventricular Remodeling and Function in Patients With Severe Aortic Stenosis
Authors: Lindman Brian R. , Arnold Suzanne V. , Madrazo José A. , Zajarias Alan , Johnson Stephanie N. , Pérez Julio E. , Mann Douglas L.
Published: February 2011

Background—

The diabetic heart exhibits increased left ventricular (LV) mass and reduced ventricular function. However, this relationship has not been studied in patients with aortic stenosis (AS), a disease process that causes LV hypertrophy and dysfunction through a distinct mechanism of pressure overload. The aim of this study was to determine how diabetes mellitus (DM) affects LV remodeling and function in patients with severe AS.

Methods and Results—

Echocardiography was performed on 114 patients with severe AS (mean aortic valve area [AVA], 0.6 cm
2
) and included measures of LV remodeling and function. Multivariable linear regression models investigated the independent effect of DM on these aspects of LV structure and function. Compared to patients without diabetes (n=60), those with diabetes (n=54) had increased LV mass and LV end-systolic and end-diastolic dimensions, and decreased LV ejection fraction (EF) and longitudinal systolic strain (all
P
<0.01). In multivariable analyses adjusting for age, sex, systolic blood pressure, AVA, body surface area, and coronary disease, DM was an independent predictor of increased LV mass (β=26 g,
P
=0.01), LV end-systolic dimension (β=0.5 cm,
P
=0.008), and LV end-diastolic dimension (β=0.3 cm,
P
=0.025). After also adjusting for LV mass, DM was associated with reduced longitudinal systolic strain (β=1.9%,
P
=0.023) and a trend toward reduced EF (β=−5%,
P
=0.09). Among patients with diabetes, insulin use (as a marker of disease severity) was associated with larger LV end-systolic dimension and worse LV function. LV mass was a strong predictor of reduced EF and systolic strain (both
P
<0.001).

Conclusions—

DM has an additive adverse effect on hypertrophic remodeling (increased LV mass and larger cavity dimensions) and is associated with reduced systolic function in patients with AS beyond known factors of pressure overload.
Pulmonary Vascular Resistance as Assessed by Bicycle Stress Echocardiography in Patients With Atrial Septal Defect Type Secundum
Authors: Van De Bruaene Alexander , La Gerche Andre , Prior David L. , Voigt Jens-Uwe , Delcroix Marion , Budts Werner
Published: February 2011

Background—

Volume overload of the pulmonary circulation in patients with atrial septal defect (ASD)-type secundum causes pulmonary vascular disease over a long period of time. This study aimed at (1) evaluating pulmonary vascular resistance (PVR) and (2) investigating the relation between PVR and exercise capacity in patients with open and closed ASD.

Methods and Results—

Forty-six patients with ASD-type secundum (18 open, 28 closed) and 20 age-matched controls were enrolled. All underwent standard and symptom-limited bicycle stress echocardiography and cardiopulmonary exercise testing. PVR was calculated as the slope of a pulmonary artery systolic pressure (PAP)-flow plot or as the ratio of PAP to cardiac output (total PVR). The slope of PAP-flow was higher in patients with open (5.1±2.2 versus 3.0±0.8 mm Hg/L per min;
P
=0.002) and closed ASD (4.0±1.7 versus 3.0±0.8 mm Hg/L per min) compared with controls. In patients with an open ASD, total PVR did not change from rest to peak exercise. In patients with a closed ASD and controls, total PVR decreased from rest to peak exercise. However, in patients with an ASD closed later in life (≥34 years, median age at repair), the slope of PAP-flow was higher (5.1±1.4 versus 3.0±0.8 mm Hg/L per min;
P
<0.0001), but total PVR did not change from rest to peak exercise. Peak oxygen consumption correlated inversely with the slope of the PAP-flow plots in patients with open (
P
=0.013) and closed ASD (
P
=0.005).

Conclusions—

In patients with an open ASD, the slope of PAP-flow was higher and total PVR did not change from rest to peak exercise. In patients with an ASD closed later in life, the slope of PAP-flow was higher and total PVR did not change from rest to peak exercise, indicating altered pulmonary hemodynamics in these patients.
Shadows From the Past: Party System Institutionalization in Asia
Authors: Hicken Allen , Martinez Kuhonta Erik
Published: February 2011

This article explains variation in levels of party system institutionalization in Asia by testing available data against several major hypotheses in the literature. The authors make three contributions to the literature on party system institutionalization. First, this study finds that historical legacies are a crucial variable affecting current levels of party system institutionalization. In particular, the immediate postwar period was the crucible from which institutionalized party systems in Asia developed. Second, the authors claim that for a significant number of institutionalized party systems, historical legacies are rooted in some element of authoritarianism, either as former authoritarian parties or as semidemocratic regimes. Third, precisely because authoritarianism has played an important role in the origins of institutionalized party systems, the authors argue that the concept of institutionalization needs to be strictly separated from the concept of democracy.
Subjective Invulnerability, Risk Behavior, and Adjustment in Early Adolescence
Authors: Hill Patrick L. , Duggan Peter M. , Lapsley Daniel K.
Published: February 2011

The current study investigated whether invulnerability manifests with adaptive and maladaptive outcomes during early adolescence. We sampled 248 (53% female; 63% Caucasian; [Formula: see text] years) early adolescents on the Adolescent Invulnerability Scale (AIS), and measures of drug use, delinquency, depressive symptoms, and mastery and coping. The AIS demonstrated a two-factor structure, which captured whether adolescents felt invulnerable to danger or psychological risks. Danger Invulnerability positively predicted delinquency and drug use. Conversely, Psychological Invulnerability negatively predicted depressive symptoms but positively predicted mastery and coping. These results suggest that felt invulnerability leads to both benefits and risks for early adolescents. Implications and future research directions are discussed.
Girls Discuss Choice of an All-Girl Middle School
Authors: Baron Emilia , Bell Nancy J. , Corson Kimberly , Kostina-Ritchey Erin , Frederick Helyne
Published: February 2011

The narrative creation of identity by young adolescents has so far been addressed mainly from an identity-in-interaction perspective, focusing attention on the multiplicity and variability of identity negotiation as adolescents interact with others, typically with peers. In contrast, a sociocultural/dialogical perspective draws attention to the importance of organization as well as variability in identity systems, and thus to the creation of personal identity stories. Our purpose is to illustrate how this perspective serves as a guide in the analysis of girls’ narratives about their decision to attend a new, all girls’ middle school. We emphasize how the stories are constituted by girls’ transactions with the sociocultural context, including societal discourses surrounding single-sex education, and we propose “specialness” as an emergent identity concept that serves to organize the identity narratives.
Promoting Diversity in the Field of Evaluation
Authors: Christie Christina A. , Vo Anne T.
Published: February 2011

In this paper we describe an evaluation training program sponsored by the Robert Wood Johnson Foundation and led by Duquesne University and OMG Center for Collaborative Learning designed to meet the challenge of developing a cadre of diverse evaluation professionals, specifically those from traditionally underrepresented or underserved communities, who have come to the field with little formal academic training in evaluation. Our interview reveals that an essential strength of this work is the collaborative relationship between an academic institution and a non-profit firm. Three additional discussion points emerged that are important learnings from this work. First, post-degree practicum training models must consider not only the training of participants, but also organizational transformation. Second, there is a genuine tension between expanding the program and the seemingly necessary responsive and intimate nature of educational programs intended to develop communities of practice, and a third and related point, are considerations of program sustainability.
Using Citation Analysis Methods to Assess the Influence of Science, Technology, Engineering, and Mathematics Education Evaluations
Authors: Greenseid Lija O. , Lawrenz Frances
Published: February 2011

This study explores the use of citation analysis methods to assess the influence of program evaluations conducted within the area of science, technology, engineering, and mathematics (STEM) education. Citation analysis is widely used within scientific research communities to measure the relative influence of scientific research enterprises and/or individual scientists, however it has not been used frequently to study the influence of evaluations. This study examines the levels, content, and networks of citations arising from products produced by four large-scale, multi-site STEM evaluations. This article explores the usefulness of citation data in understanding the impact of evaluations and finds that citation analysis methods provide information to help understand, to a limited extent, the influence of large-scale, multi-site program evaluations on the fields of STEM education and evaluation.
Acute Neurological Issues in Pregnancy and the Peripartum
Authors: Hosley Catherine M. , McCullough Louise D.
Published: February 2011

Acute neurological diseases requiring hospitalization are relatively rare in women of childbearing age. However, during pregnancy and the postpartum period, several diseases increase in prevalence. Some are unique to the pregnant/postpartum state including preeclampsia and delivery-associated neuropathies. Others, although indirectly related to pregnancy, such as cerebral venous thrombosis, ischemic stroke, and intracerebral hemorrhage, increase in frequency and carry considerable risk of morbidity and mortality. In addition, treatment options are often limited. This review discusses the diseases more commonly seen during pregnancy and the postpartum period, with a focus on emergent neurological diseases and their management. Interventional therapies will also be discussed.
Unique progenitors in mouse lymph node develop into CD127+ NK cells: thymus-dependent and thymus-independent pathways
Authors: Luther Claudia , Warner Kathrin , Takei Fumio
Published: February 2011

A subset of natural killer (NK) cells in normal mouse lymph node (LN) expresses CD127 (IL-7 receptor-α chain) and is thought to derive from the thymus. However, CD127+ NK cells are found in the LN of athymic mice. Therefore, the origin of CD127+ NK cells in the LN is unclear. Here, we have identified unique NK-cell progenitors (NKPs) in the LN that express the pan-NK cell marker CD49b and CD127 but lack CD122 and lineage markers. The LN NKPs develop in vitro into CD127+ NK cells that display natural cytotoxicity and cytokine production capacity. They also become CD127+ NK cells in lymphopenic mice that received a transplant. LN NKPs can be divided into stem cell antigen-1 (Sca-1)hi and Sca-1lo subsets. The latter comprise ∼ 60% of LN NKPs in normal mouse and < 10% of athymic mouse LN NKPs. Whereas both Sca-1hi and Sca-1lo NKPs develop into CD127+ NK cells in vitro, only those derived from Sca-1lo LN NKPs have rearranged TCRγ genes. Thus, CD127+ NK cells in the LN seem to be generated, at least in part, from both thymus-dependent Sca-1lo and thymus-independent Sca-1hi LN NKPs.
The EMT regulator Zeb2/Sip1 is essential for murine embryonic hematopoietic stem/progenitor cell differentiation and mobilization
Authors: Goossens Steven , Janzen Viktor , Bartunkova Sonia , Yokomizo Tomomasa , Drogat Benjamin , Crisan Mihaela , Haigh Katharina , Seuntjens Eve , Umans Lieve , Riedt Tamara , Bogaert Pieter , Haenebalcke Lieven , Berx Geert , Dzierzak Elaine , Huylebroeck Danny , Haigh Jody J.
Published: February 2011

Zeb2 (Sip1/Zfhx1b) is a member of the zinc-finger E-box–binding (ZEB) family of transcriptional repressors previously demonstrated to regulate epithelial-to-mesenchymal transition (EMT) processes during embryogenesis and tumor progression. We found high Zeb2 mRNA expression levels in HSCs and hematopoietic progenitor cells (HPCs), and examined Zeb2 function in hematopoiesis through a conditional deletion approach using the Tie2-Cre and Vav-iCre recombination mouse lines. Detailed cellular analysis demonstrated that Zeb2 is dispensable for hematopoietic cluster and HSC formation in the aorta-gonadomesonephros region of the embryo, but is essential for normal HSC/HPC differentiation. In addition, Zeb2-deficient HSCs/HPCs fail to properly colonize the fetal liver and/or bone marrow and show enhanced adhesive properties associated with increased β1 integrin and Cxcr4 expression. Moreover, deletion of Zeb2 resulted in embryonic (Tie2-Cre) and perinatal (Vav-icre) lethality due to severe cephalic hemorrhaging and decreased levels of angiopoietin-1 and, subsequently, improper pericyte coverage of the cephalic vasculature. These results reveal essential roles for Zeb2 in embryonic hematopoiesis and are suggestive of a role for Zeb2 in hematopoietic-related pathologies in the adult.
The E3 ubiquitin ligase RAD18 regulates ubiquitylation and chromatin loading of FANCD2 and FANCI
Authors: Williams Stacy A. , Longerich Simonne , Sung Patrick , Vaziri Cyrus , Kupfer Gary M.
Published: February 2011

Fanconi anemia (FA) is a rare genetic disorder characterized by bone marrow failure, congenital abnormalities, and an increased risk for cancer and leukemia. Components of the FA-BRCA pathway are thought to function in the repair of DNA interstrand cross-links. Central to this pathway is the monoubiquitylation and chromatin localization of 2 FA proteins, FA complementation group D2 (FANCD2) and FANCI. In the present study, we show that RAD18 binds FANCD2 and is required for efficient monoubiquitylation and chromatin localization of both FANCD2 and FANCI. Human RAD18-knockout cells display increased sensitivity to mitomycin C and a delay in FANCD2 foci formation compared with their wild-type counterparts. In addition, RAD18-knockout cells display a unique lack of FANCD2 and FANCI localization to chromatin in exponentially growing cells. FANCD2 ubiquitylation is normal in cells containing a ubiquitylation-resistant form of proliferating cell nuclear antigen, and chromatin loading of FA core complex proteins appears normal in RAD18-knockout cells. Mutation of the RING domain of RAD18 ablates the interaction with and chromatin loading of FANCD2. These data suggest a key role for the E3 ligase activity of RAD18 in the recruitment of FANCD2 and FANCI to chromatin and the events leading to their ubiquitylation during S phase.
Doxorubicin, vinblastine, and gemcitabine (CALGB 50203) for stage I/II nonbulky Hodgkin lymphoma: pretreatment prognostic factors and interim PET
Authors: Straus David J. , Johnson Jeffrey L. , LaCasce Ann S. , Bartlett Nancy L. , Kostakoglu Lale , Hsi Eric D. , Schöder Heiko , Hall Nathan C. , Jung Sin-Ho , Canellos George P. , Schwartz Lawrence H. , Takvorian Ronald W. , Juweid Malik E. , Cheson Bruce D. ,   for Cancer and Leukemia Group B
Published: February 2011

To reduce doxorubicin, bleomycin, vinblastine and dacarbazine toxicity, the Cancer and Leukemia Group B conducted a phase 2 trial of doxorubicin, vinblastine, and gemcitabine for newly diagnosed, nonbulky stages I and II Hodgkin lymphoma. Ninety-nine assessable patients received 6 cycles of doxorubicin 25 mg/m2, vinblastine 6 mg/m2, and gemcitabine 800 mg/m2 (1000 mg/m2 in first 6) on days 1 and 15 every 28 days. Computed tomography (CT) and positron emission tomography (PET) were performed before and after 2 and 6 cycles. Complete remission (CR)/CR unconfirmed was achieved in 72 of 99 patients (72.7%) and partial remission in 24 of 99 patients (24.2%). The CR rate was 81% when using PET criteria. Two patients have died of Hodgkin lymphoma progression. Median follow-up for nonprogressing patients is 3.3 years. The progression-free survival (PFS) at 3 years was 77% (95% confidence interval, 68%-84%). The relapse rate was less than 10% for patients with favorable prognostic factors. The 2-year PFS for cycle 2 PET-negative and -positive patients was 88% and 54%, respectively (P = .0009), compared with 89% and 27% for cycle 6 PET-negative and -positive patients (P = .0001). Although the CR rate and PFS were lower than anticipated, patients with favorable prognostic features had a low rate of relapse. Cycle 2 PET and cycle 6 PET were predictive of PFS. This clinical trial is registered at www.clinicaltrials.gov as #NCT00086801.
Tmprss6 is a genetic modifier of the Hfe-hemochromatosis phenotype in mice
Authors: Finberg Karin E. , Whittlesey Rebecca L. , Andrews Nancy C.
Published: February 2011

The hereditary hemochromatosis protein HFE promotes the expression of hepcidin, a circulating hormone produced by the liver that inhibits dietary iron absorption and macrophage iron release. HFE mutations are associated with impaired hepatic bone morphogenetic protein (BMP)/SMAD signaling for hepcidin production. TMPRSS6, a transmembrane serine protease mutated in iron-refractory iron deficiency anemia, inhibits hepcidin expression by dampening BMP/SMAD signaling. In the present study, we used genetic approaches in mice to examine the relationship between Hfe and Tmprss6 in the regulation of systemic iron homeostasis. Heterozygous loss of Tmprss6 in Hfe−/− mice reduced systemic iron overload, whereas homozygous loss caused systemic iron deficiency and elevated hepatic expression of hepcidin and other Bmp/Smad target genes. In contrast, neither genetic loss of Hfe nor hepatic Hfe overexpression modulated the hepcidin elevation and systemic iron deficiency of Tmprss6−/− mice. These results indicate that genetic loss of Tmprss6 increases Bmp/Smad signaling in an Hfe-independent manner that can restore Bmp/Smad signaling in Hfe−/− mice. Furthermore, these results suggest that natural genetic variation in the human ortholog TMPRSS6 might modify the clinical penetrance of HFE-associated hereditary hemochromatosis, raising the possibility that pharmacologic inhibition of TMPRSS6 could attenuate iron loading in this disorder.
Causal relationship between hyperfibrinogenemia, thrombosis, and resistance to thrombolysis in mice
Authors: Machlus Kellie R. , Cardenas Jessica C. , Church Frank C. , Wolberg Alisa S.
Published: February 2011

Epidemiologic studies have correlated elevated plasma fibrinogen (hyperfibrinogenemia) with risk of cardiovascular disease and arterial and venous thrombosis. However, it is unknown whether hyperfibrinogenemia is merely a biomarker of the proinflammatory disease state or is a causative mechanism in the etiology. We raised plasma fibrinogen levels in mice via intravenous infusion and induced thrombosis by ferric chloride application to the carotid artery (high shear) or saphenous vein (lower shear); hyperfibrinogenemia significantly shortened the time to occlusion in both models. Using immunohistochemistry, turbidity, confocal microscopy, and elastometry of clots produced in cell and tissue factor-initiated models of thrombosis, we show that hyperfibrinogenemia increased thrombus fibrin content, promoted faster fibrin formation, and increased fibrin network density, strength, and stability. Hyperfibrinogenemia also increased thrombus resistance to tenecteplase-induced thrombolysis in vivo. These data indicate that hyperfibrinogenemia directly promotes thrombosis and thrombolysis resistance and does so via enhanced fibrin formation and stability. These findings strongly suggest a causative role for hyperfibrinogenemia in acute thrombosis and have significant implications for thrombolytic therapy. Plasma fibrinogen levels may be used to identify patients at risk for thrombosis and inform thrombolytic administration for treating acute thrombosis/thromboembolism.
Essential role for Ptpn11 in survival of hematopoietic stem and progenitor cells
Authors: Chan Gordon , Cheung Laurene S. , Yang Wentian , Milyavsky Michael , Sanders Ashley D. , Gu Shengqing , Hong Wan Xing , Liu Aurora X. , Wang Xiaonan , Barbara Mary , Sharma Tarun , Gavin Joehleen , Kutok Jeffery L. , Iscove Norman N. , Shannon Kevin M. , Dick John E. , Neel Benjamin G. , Braun Benjamin S.
Published: February 2011

Src homology 2 domain-containing phosphatase 2 (Shp2), encoded by Ptpn11, is a member of the nonreceptor protein-tyrosine phosphatase family, and functions in cell survival, proliferation, migration, and differentiation in many tissues. Here we report that loss of Ptpn11 in murine hematopoietic cells leads to bone marrow aplasia and lethality. Mutant mice show rapid loss of hematopoietic stem cells (HSCs) and immature progenitors of all hematopoietic lineages in a gene dosage-dependent and cell-autonomous manner. Ptpn11-deficient HSCs and progenitors undergo apoptosis concomitant with increased Noxa expression. Mutant HSCs/progenitors also show defective Erk and Akt activation in response to stem cell factor and diminished thrombopoietin-evoked Erk activation. Activated Kras alleviates the Ptpn11 requirement for colony formation by progenitors and cytokine/growth factor responsiveness of HSCs, indicating that Ras is functionally downstream of Shp2 in these cells. Thus, Shp2 plays a critical role in controlling the survival and maintenance of HSCs and immature progenitors in vivo.
Transmission of Beauveria bassiana from male to female Aedes aegypti mosquitoes
Authors: García-Munguía Alberto M , Garza-Hernández Javier A , Rebollar-Tellez Eduardo A , Rodríguez-Pérez Mario A , Reyes-Villanueva Filiberto
Published: February 2011

Background

Resistance to chemical insecticides plus high morbidity rates have lead to rising interest in fungi as candidates for biocontrol agents of mosquito vectors. In most studies fungal infections have been induced by exposure of mosquitoes to various surfaces treated with conidia. In the present study eight Mexican strains of Beauveria bassiana were assessed against Aedes aegypti by direct exposure of females to 6 × 108 conidia ml -1 on a filter paper, afterwards, the transmission of the least and most virulent isolates was evaluated by mating behavior from virgin, fungus-contaminated male to females, to examine this ethological pattern as a new approach to deliver conidia against the dengue vector.

Methods

In an exposure chamber with a filter paper impregnated with 6 × 108 conidia ml -1 of the least and most virulent strains of B. bassiana, 6-8 day old males of A. aegypti were exposed for 48 hours, and then transferred individually (each one was a replicate) to another chamber and confined with twenty healthy females of the same age. Clean males were used in controls. Survival, infection by true mating (insemination) or by mating attempts (no insemination) and fecundity were daily registered until the death of last female. Data analysis was conducted with proc glm for unbalanced experiments and means were separated with the Ryan test with SAS.

Results

All strains were highly virulent with LT50 ranging from 2.70 (± 0.29) to 5.33 (± 0.53) days. However the most (Bb-CBG2) and least virulent (Bb-CBG4) isolates were also transmitted by mating behavior; both killed 78-90% of females in 15 days after being confined with males that had previously been exposed for 48 hours to fungi. Of these mortality rates, 23 and 38% respectively, were infections acquired by copulations where insemination occurred. The LT50 for sexually-infected females were 7.92 (± 0.46) and 8.82 (± 0.45) days for both strains, while the one in control was 13.92 (± 0.58). Likewise, fecundity decreased by 95% and 60% for both Bb-CBG2 and Bb-CBG4 isolates in comparison with control. The role of mating attempts in this delivery procedure of B. bassiana is discussed.

Conclusions

This is the first report about transmission of B. bassiana by mating behavior from virgin, fungus-contaminated males to females in A. aegypti. Fungal infections acquired by this route (autodissemination) infringed high mortality rates (90%) in mated or approached females. However, prior to releasing virgin, fungus-contaminated males to spread B. basasiana among females of A. aegypti, this novel alternative needs further investigations.
Antitumor activity of mixed heat shock protein/peptide vaccine and cyclophosphamide plus interleukin-12 in mice sarcoma
Authors: Guo Quan-Yi , Yuan Mei , Peng Jiang , Cui Xue-Mei , Song Ge , Sui Xiang , Lu Shi-Bi
Published: February 2011

Background

The immune factors heat shock protein (HSP)/peptides (HSP/Ps) can induce both adaptive and innate immune responses. Treatment with HSP/Ps in cancer cell-bearing mice and cancer patients revealed antitumor immune activity. We aimed to develop immunotherapy strategies by vaccination with a mixture of HSP/Ps (mHSP/Ps, HSP60, HSP70, Gp96 and HSP110) enhanced with cyclophosphamide (CY) and interleukin-12 (IL-12).

Methods

We extracted mHSP/Ps from the mouse sarcoma cell line S180 using chromatography. The identity of proteins in this mHSP/Ps was assayed using SDS-PAGE and Western blot analysis with antibodies specific to various HSPs. BALB/C mice bearing S180 cells were vaccinated with mHSP/Ps ×3, then were injected intraperitoneally with low-dose CY and subcutaneously with IL-12, 100 μg/day, ×5. After vaccination, T lymphocytes in the peripheral blood were analyzed using FACScan and Cytotoxicity (CTL) was analyzed using lactate dehydrogenase assay. ELISPOT assay was used to evaluate interferon γ (IFN-γ), and immune cell infiltration in tumors was examined in the sections of tumor specimen.

Results

In mice vaccinated with enhanced vaccine (mHSP/Ps and CY plus IL-12), 80% showed tumor regression and long-term survival, and tumor growth inhibition rate was 82.3% (30 days), all controls died within 40 days. After vaccination, lymphocytes and polymorphonuclear leukocytes infiltrated into the tumors of treated animals, but no leukocytes infiltrated into the tumors of control mice. The proportions of natural killer cells, CD8+, and interferon-γ-secreting cells were all increased in the immune group, and tumor-specific cytotoxic T lymphocyte activity was increased.

Conclusions

In this mice tumor model, vaccination with mHSP/Ps combined with low-dose CY plus IL-12 induced an immunologic response and a marked antitumor response to autologous tumors. The regimen may be a promising therapeutic agent against tumors.
Interference of gadolinium on the measurement of selenium in human serum by inductively coupled plasma-quadrupole mass spectrometry
Authors: Walter Alan , Nelms Simon , Harrington Chris F , Taylor Andrew
Published: February 2011

Selenium is an important clinical biomarker of nutritional status; however, the occurrence of gadolinium in a patient's serum as a result of the contrast agents used during magnetic resonance imaging investigations, results in a significant positive bias in its measurement by inductively coupled plasma-quadrupole mass spectrometry.
Liquid chromatography-tandem mass spectrometry method for the measurement of serum mevalonic acid: a novel marker of hydroxymethylglutaryl coenzyme A reductase inhibition by statins
Authors: Waldron Jenna , Webster Craig
Published: February 2011

Background

Mevalonic acid (MVA) is synthesized at an early and rate-limiting step in the biosynthesis of cholesterol by the enzyme hydroxymethylglutaryl coenzyme A (HMG-CoA) reductase, and is a useful measure of statin efficacy or treatment. A liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for the measurement of serum MVA has been developed.

Methods

Following the in vitro conversion of MVA to mevalonic acid lactone (MVAL) in the serum, MVAL and a deuterated internal standard were extracted using an online solid-phase extraction procedure. Chromatographic separation was achieved using a Luna PFP column (Phenomenex), with enhanced selectivity and improved resolution for polar compounds. A gradient system was used, with mobile phase comprising methanol and water (5 mmol/L ammonium formate buffer, pH 2.5). Analysis was performed using an API 5000 tandem mass spectrometer (Applied Biosystems) in positive electrospray ionization mode.

Results

The method showed excellent recoveries (98 ± 8%) and imprecision (intra-assay coefficient of variation of 2.2% [6.5 ng/mL] and 2.6% [10.5 ng/mL], and inter-assay coefficient of variation of 9% [10.5 ng/mL]). The assay provides a calibration range up to 50 ng/mL with a limit of detection at 0.1 ng/mL.

Conclusion

A simple, rapid and analytically specific method has been developed for the measurement of serum MVA, in the form of MVAL. The high analytical sensitivity of the method allows for accurate quantitation of MVAL in serum samples, both at the endogenous levels found in healthy individuals and in statin-treated patients where normal levels are expected to be greatly reduced through the inhibition of HMG-CoA reductase.
Evaluation of the interference due to haemoglobin, bilirubin and lipids on Immulite 2500 assays: a practical approach
Authors: Steen Gerard , Klerk Alex , Laan Kees van der , Eppens Elaine F
Published: February 2011

Background

Interfering substances such as haemoglobin, bilirubin and lipids in a sample may lead to wrong interpretation of immunoassay results by the clinician. In general, there has been minor attention to these interferences on immunoassays, whereas these effects on chemical assays are frequently described. Information about interferences by haemoglobin, bilirubin and lipids on the Siemens Immulite 2500 assays in the instructions for use is falling short.

Methods

Interferents in patient samples can be measured reliably in a semi-quantitative way on most chemistry analysers and can be expressed in haemolysis-, icterus- and lipaemia-indices. As the Immulite 2500 cannot perform such measurements, samples are normally analysed without testing for the presence of interferents. Therefore, a study was carried out to examine these interferences on 24 Immulite 2500 assays. Samples were spiked with increasing concentrations of either haemoglobin, bilirubin or lipids. The haemolysis-, icterus- and lipaemia-indices were measured on a Synchron DxC 800 analyser.

Results

Based on analytical imprecision and intraindividual biological variation of each analyte, cut-off indices above which clinically significant interference exists were determined. We found clinically significant interference due to haemoglobin on ferritin and folate, by bilirubin on oestradiol and testosterone and by lipids on testosterone.

Conclusions

Introducing cut-off indices prevents reporting of wrong Immulite 2500 results due to interference. Our results are applicable in laboratories using any chemistry analyser capable of reporting semi-quantitative concentrations of interferents.
The implications of chronic pain models for rehabilitation of distal radius fracture
Authors: Mehta Saurabh , MacDermid Joy , Tremblay Mary
Published: February 2011

Distal radius fracture (DRF) is the most common fracture and usually occurs as a result of a fall. Most patients recover following DRF with minimal residual pain or disability; however, a small subset of patients continue to experience pain and disability even one year after the injury. Currently, there are no practice guidelines for early identification and treatment of patients who are potentially at greater risk of developing these adverse outcomes. As a result, hand therapy management of patients following DRF does not incorporate screening of these at-risk patients. The objective of this paper is to apply constructs from learned helplessness and cognitive-behavioural models of chronic pain in assessing the psychosocial risk profile of patients following DRF. We have also integrated key findings derived from studies addressing personal and life-style factors in assessing this risk profile. This framework is proposed as a basis to categorize patients as higher or lower psychosocial risk for developing chronic pain and disability following DRF. We outline a model depicting the RACE approach (Reducing pain, Activating, Cognitive reshaping, Empowering) towards the management of patients following DRF. The model suggests that patients with minimal psychosocial risk factors are managed based on their injury profile and those with higher psychosocial risk are treated with the risk-based RACE approach. Using a biopsychosocial RACE approach to prognosis and treatment, hand therapy intervention can be customized for patients recovering from DRF. In future, researchers can conduct clinical trials to compare the RACE-based treatment approach to routine hand therapy in mitigating the risk of chronic pain and disability in patients with elevated risk profile for adverse outcomes following DRF.
A study of the effect of desensitization on hyperaesthesia in the hand and upper extremity after injury or surgery
Authors: Göransson Ingrid , Cederlund Ragnhild
Published: February 2011

Introduction

Hyperaesthesia is a common problem in hand rehabilitation. A desensitization programme has successfully been used in our occupational therapy department for patients with hyperaesthesia in the hand or upper extremity. Our objectives were to investigate self-perceived pain/discomfort at rest, with use or touch, size of sensitive skin area and occupational performance in daily occupations for patients with hyperaesthesia before and after a six-week desensitization programme.

Methods

Thirty-nine patients with pain/discomfort at or near scars from hand injury or surgery participated in the study. The patients were recruited consecutively to the study. They were instructed to massage the sensitive skin area three times a day with a piece of texture until numbness occurred (2–5 minutes). The textures were upgraded continuously. A study-specific questionnaire was developed including demographic data and questions concerning perceived pain/discomfort. Pain/discomfort was assessed with a visual analogue scale 0–100. The size of the sensitive skin area was assessed in mm2. Occupational performance was assessed with the Canadian Occupational Performance Measure.

Results

The results showed statistically significant improvements ( P ≤ 0.001) in all areas after six weeks with reduced pain/discomfort at rest and with use or touch, a decreased size of the sensitive skin area and a higher occupational performance in daily occupations.

Discussion

The results indicate the importance of paying attention to sensory disturbances during rehabilitation. The cost for such a programme is low. Further research is needed to confirm the results in a larger sample using a control group, and in comparison with spontaneous healing and other treatment modalities.