bluuebunny/crossref_metadata_2025 huggingface dataset:
Cancer Cell–Associated MT1-MMP Promotes Blood Vessel Invasion and Distant Metastasis in Triple-Negative Mammary Tumors
Authors: Perentes Jean Y. , Kirkpatrick Nathaniel D. , Nagano Satoshi , Smith Eve Y. , Shaver Christine M. , Sgroi Dennis , Garkavtsev Igor , Munn Lance L. , Jain Rakesh K. , Boucher Yves
Published: May 2011

Functional roles for the cancer cell–associated membrane type I matrix metalloproteinase (MT1-MMP) during early steps of the metastatic cascade in primary tumors remain unresolved. In an effort to determine its significance, we determined the in vivo effects of RNAi-mediated downregulation in mammary cancer cells on the migration, blood and lymphatic vessel invasion (LVI), and lymph node and lung metastasis. We also correlated the expression of cancer cell MT1-MMP with blood vessel invasion (BVI) in 102 breast cancer biopsies. MT1-MMP downregulation in cancer cells decreased lung metastasis without affecting primary tumor growth. The inhibition of lung metastasis correlated with reduced cancer cell migration and BVI. Furthermore, cancer cell–expressed MT1-MMP upregulated the expression of MT1-MMP in vascular endothelial cells, but did not affect MT1-MMP expression in lymphatic endothelial cells, LVI, or lymph node metastasis. Of clinical importance, we observed that elevated MT1-MMP expression correlated with BVI in biopsies from triple-negative breast cancers (TNBC), which have a poor prognosis and high incidence of distant metastasis, relative to other breast cancer subtypes. Together, our findings established that MT1-MMP activity in breast tumors is essential for BVI, but not LVI, and that MT1-MMP should be further explored as a predictor and therapeutic target of hematogenous metastasis in TNBC patients. Cancer Res; 71(13); 4527–38. ©2011 AACR.
RECQL1 and WRN Proteins Are Potential Therapeutic Targets in Head and Neck Squamous Cell Carcinoma
Authors: Arai Akihito , Chano Tokuhiro , Futami Kazunobu , Furuichi Yasuhiro , Ikebuchi Kaichiro , Inui Takuma , Tameno Hitosuke , Ochi Yasuko , Shimada Taketoshi , Hisa Yasuo , Okabe Hidetoshi
Published: May 2011

RECQL1 and WRN proteins are RecQ DNA helicases that participate in suppression of DNA hyper-recombination and repair. In this study, we report evidence supporting their candidacy as cancer therapeutic targets. In hypopharyngeal carcinomas, which have the worst prognosis among head and neck squamous cell carcinomas (HNSCC) that are rapidly rising in incidence, we found that RECQL1 and WRN proteins are highly expressed and that siRNA-mediated silencing of either gene suppressed carcinoma cell growth in vitro. Similarly, siRNA administration in a murine xenograft model of hypopharyngeal carcinoma markedly inhibited tumor growth. Moreover, combining either siRNA with cis-platinum (II) diammine dichloride significantly augmented the in vivo anticancer effects of this drug that is used commonly in HNSCC treatment. Notably, we observed no recurrence of some tumors following siRNA treatment in this model. Our findings offer a preclinical proof of concept for RECQL1 and WRN proteins as novel therapeutic targets to treat aggressive HNSCC and perhaps other cancers. Cancer Res; 71(13); 4598–607. ©2011 AACR.
Trastuzumab and Pertuzumab Produce Changes in Morphology and Estrogen Receptor Signaling in Ovarian Cancer Xenografts Revealing New Treatment Strategies
Authors: Faratian Dana , Zweemer Annelien J.M. , Nagumo Yoko , Sims Andrew H. , Muir Morwenna , Dodds Michael , Mullen Peter , Um Inhwa , Kay Charlene , Hasmann Max , Harrison David J. , Langdon Simon P.
Published: May 2011

Purpose: The aim of this study was to investigate the antitumor effects of HER2-directed combination therapy in ovarian cancer xenograft models to evaluate their potential. The combinations of trastuzumab and pertuzumab, and trastuzumab and aromatase inhibitor therapy were investigated.

Experimental Design: The effects of trastuzumab, pertuzumab, and letrozole on growth response, apoptosis, morphology, and gene and protein expression were evaluated in the SKOV3 ovarian cancer cell line xenograft and a panel of five human ovarian xenografts derived directly from clinical specimens.

Results: The combination of HER2-directed antibodies showed enhanced antitumor activity compared with single antibody therapy in the SKOV3 xenograft model. Apoptosis, morphology, and estrogen-regulated gene expression were modulated by these antibodies in both spatial and temporal manners. A panel of ovarian cancer xenografts showed differential growth responses to the combination of trastuzumab and pertuzumab. High HER2 expression and increasing HER3 protein expression on treatment were associated with growth response. In trastuzumab-treated SKOV3 tumors, there was a change in tumor morphology, with a reduction in frequency of estrogen receptor alpha (ERα)-negative clear cell areas. Trastuzumab, but not pertuzumab, increased expression of ERα in SKOV3 xenografts when analyzed by quantitative immunofluorescence. ERα and downstream signaling targets were modulated by trastuzumab alone and in combination. Trastuzumab enhanced the responsiveness of SKOV3 xenografts to letrozole when given in combination.

Conclusions: These data suggest that trastuzumab in combination with pertuzumab could be an effective approach in high HER2-expressing ovarian cancers and could also enhance sensitivity to endocrine therapy in ERα-positive ovarian cancer. Clin Cancer Res; 17(13); 4451–61. ©2011 AACR.
KL1 Internal Repeat Mediates Klotho Tumor Suppressor Activities and Inhibits bFGF and IGF-I Signaling in Pancreatic Cancer
Authors: Abramovitz Lilach , Rubinek Tamar , Ligumsky Hagai , Bose Shikha , Barshack Iris , Avivi Camila , Kaufman Bella , Wolf Ido
Published: May 2011

Purpose: Klotho is a transmembrane protein which can be shed, act as a circulating hormone and modulate the insulin-like growth factor (IGF)-I and the fibroblast growth factor (FGF) pathways. We have recently identified klotho as a tumor suppressor in breast cancer. Klotho is expressed in the normal pancreas and both the IGF-I and FGF pathways are involved in pancreatic cancer development. We, therefore, undertook to study the expression and activity of klotho in pancreatic cancer.

Experimental Design: Klotho expression was studied using immunohistochemistry and quantitative RT-PCR. Effects of klotho on cell growth were assessed in the pancreatic cancer cells Panc1, MiaPaCa2, and Colo357, using colony and MTT assays and xenograft models. Signaling pathway activity was measured by Western blotting.

Results: Klotho expression is downregulated in pancreatic adenocarcinoma. Overexpression of klotho, or treatment with soluble klotho, reduced growth of pancreatic cancer cells in vitro and in vivo, and inhibited activation of the IGF-I and the bFGF pathways. KL1 is a klotho subdomain formed by cleavage or alternative splicing. Compared with the full-length protein, KL1 showed similar growth inhibitory activity but did not promote FGF23 signaling. Thus, its administration to mice showed favorable safety profile.

Conclusions: These studies indicate klotho as a potential tumor suppressor in pancreatic cancer, and suggest, for the first time, that klotho tumor suppressive activities are mediated through its KL1 domain. These results suggest the use of klotho or KL1 as potential strategy for the development of novel therapeutic interventions for pancreatic cancer. Clin Cancer Res; 17(13); 4254–66. ©2011 AACR.
EFEMP1 as a Novel DNA Methylation Marker for Prostate Cancer: Array-Based DNA Methylation and Expression Profiling
Authors: Kim Yong-June , Yoon Hyung-Yoon , Kim Seon-Kyu , Kim Young-Won , Kim Eun-Jung , Kim Isaac Yi , Kim Wun-Jae
Published: May 2011

Purpose: Abnormal DNA methylation is associated with many human cancers. The aim of the present study was to identify novel methylation markers in prostate cancer (PCa) by microarray analysis and to test whether these markers could discriminate normal and PCa cells.

Experimental Design: Microarray-based DNA methylation and gene expression profiling was carried out using a panel of PCa cell lines and a control normal prostate cell line. The methylation status of candidate genes in prostate cell lines was confirmed by real-time reverse transcriptase-PCR, bisulfite sequencing analysis, and treatment with a demethylation agent. DNA methylation and gene expression analysis in 203 human prostate specimens, including 106 PCa and 97 benign prostate hyperplasia (BPH), were carried out. Further validation using microarray gene expression data from the Gene Expression Omnibus (GEO) was carried out.

Results: Epidermal growth factor–containing fibulin-like extracellular matrix protein 1 (EFEMP1) was identified as a lead candidate methylation marker for PCa. The gene expression level of EFEMP1 was significantly higher in tissue samples from patients with BPH than in those with PCa (P < 0.001). The sensitivity and specificity of EFEMP1 methylation status in discriminating between PCa and BPH reached 95.3% (101 of 106) and 86.6% (84 of 97), respectively. From the GEO data set, we confirmed that the expression level of EFEMP1 was significantly different between PCa and BPH.

Conclusion: Genome-wide characterization of DNA methylation profiles enabled the identification of EFEMP1 aberrant methylation patterns in PCa. EFEMP1 might be a useful indicator for the detection of PCa. Clin Cancer Res; 17(13); 4523–30. ©2011 AACR.
Response of Human Prostate Cancer Cells and Tumors to Combining PARP Inhibition with Ionizing Radiation
Authors: Barreto-Andrade Juan Camilo , Efimova Elena V. , Mauceri Helena J. , Beckett Michael A. , Sutton Harold G. , Darga Thomas E. , Vokes Everett E. , Posner Mitchell C. , Kron Stephen J. , Weichselbaum Ralph R.
Published: May 2011

Radiation therapy remains a promising modality for curative treatment of localized prostate cancer, but dose-limiting toxicities significantly limit its effectiveness. Agents that enhance efficacy at lower radiation doses might have considerable value in increasing tumor control without compromising organ function. Here, we tested the hypothesis that the PARP inhibitor ABT-888 (veliparib) can enhance the response of prostate cancer cells and tumors to ionizing radiation (IR). Following exposure of DU-145 and PC-3 prostate cancer cell lines to the combination of 10 μmol/L ABT-888 and 6 Gy, we observed similar persistence between both cell lines of DNA damage foci and in vitro radiosensitization. We have previously observed that persistent DNA damage foci formed after ABT-888 plus IR efficiently promote accelerated cell senescence, but only PC-3 cells displayed the expected senescent response of G2–M arrest, induction of p21 and β-galactosidase expression, and accumulation as large flat cells. In turn, combining ABT-888 with 6 Gy resulted in delayed tumor regrowth compared with either agent alone only in PC-3 xenograft tumors, whereas DU-145 tumors continued to grow. By 7 days after treatment with ABT-888 plus IR, PC-3 tumors contained abundant senescent cells displaying persistent DNA damage foci, but no evidence of senescence was noted in the DU-145 tumors. That equivalent radiosensitization by ABT-888 plus IR in vitro failed to predict comparable results with tumors in vivo suggests that the efficacy of PARP inhibitors may partially depend on a competent senescence response to accumulated DNA damage. Mol Cancer Ther; 10(7); 1185–93. ©2011 AACR.
Phase I Trial of Hepatic Arterial Infusion of Nanoparticle Albumin–Bound Paclitaxel: Toxicity, Pharmacokinetics, and Activity
Authors: Fu Siqing , Naing Aung , Moulder Stacy L. , Culotta Kirk S. , Madoff David C. , Ng Chaan S. , Madden Timothy L. , Falchook Gerald S. , Hong David S. , Kurzrock Razelle
Published: May 2011

Because liver involvement in patients with metastatic cancer has limited options and poor outcomes, we conducted a phase I study to determine the safety, activity, and pharmacokinetic characteristics of hepatic arterial infusion of nanoparticle albumin–bound paclitaxel (HAI nab-paclitaxel). Cohorts of three patients having predominant hepatic metastases received HAI nab-paclitaxel at three dose levels (180, 220, and 260 mg/m2, respectively) infused for more than 1 hour every 3 weeks (3 + 3 design). Some patients participated in comparative pharmacokinetic studies (i.v. vs. HAI), receiving their first course i.v., to determine peak concentrations and effect of first-pass hepatic extraction compared with subsequent courses administered by HAI. The highest dose level was expanded to determine the safety and activity of HAI nab-paclitaxel. Thirty-eight patients were treated. There were no dose-limiting toxicities at doses up to 260 mg/m2. Common adverse events included alopecia, fatigue, myelosuppresion, nausea, and vomiting. Three patients had stable disease for 4 or more months and 2 patients (1 of 12 with breast cancer and 1 of 1 with cervical cancer) achieved a partial response lasting for 5 and 15 months, respectively. Peak concentrations were lower (∼50%) with greater hepatic extraction of drug (∼42%) following HAI than i.v. infusion based on area under the curve comparison of drug exposure. HAI nab-paclitaxel showed partial hepatic extraction. At doses 260 mg/m2 or less given for 1 hour every 3 weeks, the treatment was well-tolerated and showed activity in advanced cancer patients with predominant liver metastases. Mol Cancer Ther; 10(7); 1300–7. ©2011 AACR.
COP35, a Cholangiocarcinoma-Binding Oligopeptide, Interacts with the Clathrin Heavy Chain Accompanied by GRP78
Authors: Kitahara Hiroe , Masumoto Junya , Parker Alan L. , Maruta Fukuto , Kubo Naoki , Shimizu Akira , Akita Noriyuki , Miwa Shiro , Kobayashi Naoya , Nakayama Jun , Miyagawa Shinichi
Published: May 2011

Cholangiocarcinoma (CCA) is a common carcinoma of the liver, and the majority of patients with CCA have a poor prognosis due to the lack of effective nonsurgical therapies in addition to its rapid progression and inoperability at the time of diagnosis. The development of novel nonsurgical therapeutics that efficiently target CCA could significantly improve the prognosis for patients presenting with CCA. Here, we describe the iterative production and characterization of a novel peptide, designated COP35 (CCA-binding oligopeptide 35), which binds selectively to human CCA, identified by bacteriophage biopanning using the intrahepatic CCA cell line RBE and the normal cholangiocyte cell line MMNK-1. COP35 was found to augment the growth inhibitory effects of 5-fluorouracil (5-FU) against RBE cells. Utilizing pull-down assay and liquid chromatography, we identify the clathrin heavy chain accompanied by GRP78/BiP as a COP35-binding partner. In summary, we identify COP35 as a possible candidate for peptide-targeted therapies for CCA. Mol Cancer Res; 9(6); 688–701. ©2011 AACR.
Differential Hypoxic Regulation of Hypoxia-Inducible Factors 1α and 2α
Authors: Lin Qun , Cong Xiangyu , Yun Zhong
Published: May 2011

The hypoxia-inducible transcription factors (HIF) 1α and HIF-2α play a critical role in cellular response to hypoxia. Elevated HIF-α expression correlates with poor patient survival in a large number of cancers. Recent evidence suggests that HIF-2α appears to be preferentially expressed in neuronal tumor cells that exhibit cancer stem cell characteristics. These observations suggest that expression of HIF-1α and HIF-2α is differentially regulated in the hypoxic tumor microenvironment. However, the underlying mechanisms remain to be fully investigated. In this study, we investigated the transcriptional regulation of HIF-1α and HIF-2α under different physiologically relevant hypoxic conditions. We found that transcription of HIF-2α was consistently increased by hypoxia, whereas transcription of HIF-1α showed variable levels of repression. Mechanistically, differential regulation of HIF-α transcription involved hypoxia-induced changes in acetylation of core histones H3 and H4 associated with the proximal promoters of the HIF-1α or HIF-2α gene. We also found that, although highly stable under acute hypoxia, HIF-1α and HIF-2α proteins become destabilized under chronic hypoxia. Our results have thus provided new mechanistic insights into the differential regulation of HIF-1α and HIF-2α by the hypoxic tumor microenvironment. These findings also suggest an important role of HIF-2α in the regulation of tumor progression under chronic hypoxia. Mol Cancer Res; 9(6); 757–65. ©2011 AACR.
Sex Hormones Alter Th1 Responses and Enhance Granuloma Formation in the Lung
Authors: Tajima Ken , Miura Kayo , Ishiwata Toshiji , Takahashi Fumiyuki , Yoshioka Masataka , Minakata Kunihiko , Murakami Akiko , Sasaki Shinichi , Iwakami Shinichiro , Annoura Takeshi , Hashimoto Muneaki , Nara Takeshi , Takahashi Kazuhisa
Published: May 2011

Background: The lung is one of the sites of granulomatous responses, which are characterized by the recruitment and organization of activated macrophages and lymphocytes. There have been several reports that have shown that some pulmonary granulomatous diseases, such as sarcoidosis and nontuberculous mycobacterial disease, are likely to be characterized by a preponderance in postmenopausal females. Although sex hormones have been shown to play an important role in the regulation of the immune system, the influence of sex hormones on pulmonary granuloma formation is still unclear. Objectives: The purpose of this study was to assess whether sex hormones are involved in granulomatous inflammation and to evaluate how sex hormones modulate this response in the lung. Methods: Ovariectomized rats were used as an experimental postmenopausal model in which chronic pulmonary granulomatous inflammation was induced by intravenous injection of complete Freund’s adjuvant. Results: Histological analysis of lung tissues demonstrated enhancement of granuloma formation in the ovariectomized group. Such enhanced granuloma formation was significantly associated with generalized Th1-biased cytokine production in the bronchoalveolar lavage fluid. Conclusion: These results indicate that sex hormones play an important role in pulmonary granuloma formation by altering the Th1 responses.
Reverse Cholesterol Transport Revisited
Authors: Brufau Gemma , Groen Albert K. , Kuipers Folkert
Published: May 2011

Reverse cholesterol transport (RCT) is usually defined as high-density lipoprotein-mediated transport of excess cholesterol from peripheral tissues, including cholesterol-laden macrophages in vessel walls, to the liver. From the liver, cholesterol can then be removed from the body via secretion into the bile for eventual disposal via the feces. According to this paradigm, high plasma high-density lipoprotein levels accelerate RCT and hence are atheroprotective. New insights in individual steps of the RCT pathway, in part derived from innovative mouse models, indicate that the classical concept of RCT may require modification.
Dietary α-Linolenic Acid Inhibits Arterial Thrombus Formation, Tissue Factor Expression, and Platelet Activation
Authors: Holy Erik W. , Forestier Marc , Richter Eva K. , Akhmedov Alexander , Leiber Florian , Camici Giovanni G. , Mocharla Pavani , Lüscher Thomas F. , Beer Jürg H. , Tanner Felix C.
Published: May 2011

Objective—

Plant-derived α-linolenic acid (ALA) may constitute an attractive cardioprotective alternative to fish-derived
n
-3 fatty acids. However, the effect of dietary ALA on arterial thrombus formation remains unknown.

Methods and Results—

Male C57Bl/6 mice were fed a high-ALA or low-ALA diet for 2 weeks. Arterial thrombus formation was delayed in mice fed a high-ALA diet compared with those on a low-ALA diet (n=7;
P
<0.005). Dietary ALA impaired platelet aggregation to collagen and thrombin (n=5;
P
<0.005) and decreased p38 mitogen-activated protein kinase activation in platelets. Dietary ALA impaired arterial tissue factor (TF) expression, TF activity, and nuclear factor-κB activity (n=7;
P
<0.05); plasma clotting times and plasma thrombin generation did not differ (n=5;
P
=not significant). In cultured human vascular smooth muscle and endothelial cells, ALA inhibited TF expression and activity (n=4;
P
<0.01). Inhibition of TF expression occurred at the transcriptional level via the mitogen-activated protein kinase p38 in smooth muscle cells and p38, extracellular signal-regulated kinases 1 and 2, and c-Jun N-terminal kinases 1 and 2 in endothelial cells.

Conclusion—

ALA impairs arterial thrombus formation, TF expression, and platelet activation and thereby represents an attractive nutritional intervention with direct dual antithrombotic effects.
Multifaceted Role of Plasminogen Activator Inhibitor-1 in Regulating Early Remodeling of Vein Bypass Grafts
Authors: Ji Yan , Strawn Tammy L. , Grunz Elizabeth A. , Stevenson Meredith J. , Lohman Alexander W. , Lawrence Daniel A. , Fay William P.
Published: May 2011

Objective—

The role of plasminogen activator inhibitor-1 (PAI-1) in vein graft (VG) remodeling is undefined. We examined the effect of PAI-1 on VG intimal hyperplasia and tested the hypothesis that PAI-1 regulates VG thrombin activity.

Methods and Results—

VGs from wild-type (WT),
Pai1
−/−
, and PAI-1-transgenic mice were implanted into WT,
Pai1
−/−
, or PAI-1-transgenic arteries. VG remodeling was assessed 4 weeks later. Intimal hyperplasia was significantly greater in PAI-1-deficient mice than in WT mice. The proliferative effect of PAI-1 deficiency was retained in vitronectin-deficient mice, suggesting that PAI-1's antiproteolytic function plays a key role in regulating intimal hyperplasia. Thrombin-induced proliferation of PAI-1-deficient venous smooth muscle cells (SMC) was significantly greater than that of WT SMC, and thrombin activity was significantly higher in PAI-1-deficient VGs than in WT VGs. Increased PAI-1 expression, which has been associated with obstructive VG disease, did not increase intimal hyperplasia.

Conclusion—

Decreased PAI-1 expression (1) promotes intimal hyperplasia by pathways that do not require vitronectin and (2) increases thrombin activity in VG. PAI-1 overexpression, although it promotes SMC migration in vitro, did not increase intimal hyperplasia. These results challenge the concept that PAI-1 drives nonthrombotic obstructive disease in VG and suggest that PAI-1's antiproteolytic function, including its antithrombin activity, inhibits intimal hyperplasia.
Genetic Susceptibility of the Arterial Wall Is an Important Determinant of Atherosclerosis in C57BL/6 and FVB/N Mouse Strains
Authors: Shim Jeong , Handberg Aase , Östergren Caroline , Falk Erling , Bentzon Jacob F.
Published: May 2011

Objective—

How genetic variations among inbred mouse strains translate into differences in atherosclerosis susceptibility is of significant interest for the development of new therapeutic strategies. The objective of the present study was to examine whether genetically controlled arterial wall properties influence atherosclerosis susceptibility in FVB/N (FVB) and C57BL/6 (B6) apolipoprotein E knockout (apoE
−/−
) mouse strains.

Methods and Results—

Common carotid artery segments from B6 apoE
−/−
, F1 apoE
−/−
, and FVB apoE
−/−
mice were transplanted to hybrid F1 apoE
−/−
mice, which can accept grafts from both parental strains without adaptive immune responses. The mice were fed a high-fat diet, and atherosclerosis was induced in the transplanted artery segments by placement of a perivascular constrictive collar. Artery segments from B6 apoE
−/−
mice developed much larger atherosclerotic lesions than artery segments from FVB or F1 apoE
−/−
mice. No differences in aortic arch atherosclerosis of the recipient mice were observed between groups.

Conclusion—

Genetically controlled factors acting at the level of the arterial wall are important determinants of atherosclerosis susceptibility in FVB apoE
−/−
and B6 apoE
−/−
mice.
Methodological Development of a Clonogenic Assay to Determine Endothelial Progenitor Cell Potential
Authors: Masuda Haruchika , Alev Cantas , Akimaru Hiroshi , Ito Rie , Shizuno Tomoko , Kobori Michiru , Horii Miki , Ishihara Toshiya , Isobe Kazuya , Isozaki Mitsuhiro , Itoh Johbu , Itoh Yoshiko , Okada Yoshinori , McIntyre Brendan A.S. , Kato Shunichi , Asahara Takayuki
Published: May 2011

The precise and conceptual insight of circulating endothelial progenitor cell (EPC) kinetics is hampered by the absence of an assay system capable of evaluating the EPC differentiation cascade. An assay system for EPC colony formation was developed to delineate circulating EPC differentiation. EPC colony-forming assay using semisolid medium and single or bulk CD133
+
cells from umbilical cord blood exhibited the formation of two types of attaching cell colonies made of small or large cells featuring endothelial lineage potential and properties, termed small EPC colony-forming units and large EPC colony-forming units, respectively. In vitro and in vivo assays of each EPC colony-forming unit cell revealed a differentiation hierarchy from small EPC to large EPC colonies, indicating a primitive EPC stage with highly proliferative activity and a definitive EPC stage with vasculogenic properties, respectively. Experimental comparison with a conventional EPC culture assay system disclosed EPC colony-forming unit cells differentiate into noncolony-forming early EPC. The fate analysis of single CD133
+
cells into the endothelial and hematopoietic lineage was achieved by combining this assay system with a hematopoietic progenitor assay and demonstrated the development of colony-forming EPC and hematopoietic progenitor cells from a single hematopoietic stem cell. EPC colony-forming assay permits the determination of circulating EPC kinetics from single or bulk cells, based on the evaluation of hierarchical EPC colony formation. This assay further enables a proper exploration of possible links between the origin of EPC and hematopoietic stem cells, representing a novel and powerful tool to investigate the molecular signaling pathways involved in EPC biology.
Informatic and Functional Approaches to Identifying a Regulatory Region for the Cardiac Sodium Channel
Authors: Atack Thomas C. , Myers Stroud Dina , Watanabe Hiroshi , Yang Tao , Hall Lynn , Hipkens Susan B. , Lowe John S. , Leake Brenda , Magnuson Mark A. , Yang Ping , Roden Dan M.
Published: May 2011

Rationale:

Although multiple lines of evidence suggest that variable expression of the cardiac sodium channel gene
SCN5A
plays a role in susceptibility to arrhythmia, little is known about its transcriptional regulation.

Objective:

We used in silico and in vitro experiments to identify possible noncoding sequences important for transcriptional regulation of
SCN5A
. The results were extended to mice in which a putative regulatory region was deleted.

Methods and Results:

We identified 92 noncoding regions highly conserved (>70%) between human and mouse
SCN5A
orthologs. Three conserved noncoding sequences (CNS) showed significant (>5-fold) activity in luciferase assays. Further in vitro studies indicated one, CNS28 in intron 1, as a potential regulatory region. Using recombinase-mediated cassette exchange (RMCE), we generated mice in which a 435–base pair region encompassing CNS28 was removed. Animals homozygous for the deletion showed significant increases in
SCN5A
transcripts, Na
V
1.5 protein abundance, and sodium current measured in isolated ventricular myocytes. ECGs revealed a significantly shorter QRS (10.7±0.2 ms in controls versus 9.7±0.2 ms in knockouts), indicating more rapid ventricular conduction. In vitro analysis of CNS28 identified a short 3′ segment within this region required for regulatory activity and including an E-box motif. Deletion of this segment reduced reporter activity to 3.6%±0.3% of baseline in CHO cells and 16%±3% in myocytes (both
P
<0.05), and mutation of individual sites in the E-box restored activity to 62%±4% and 57%±2% of baseline in CHO cells and myocytes, respectively (both
P
<0.05).

Conclusions:

These findings establish that regulation of cardiac sodium channel expression modulates channel function in vivo, and identify a noncoding region underlying this regulation.
Calcium Regulates Key Components of Vascular Smooth Muscle Cell–Derived Matrix Vesicles to Enhance Mineralization
Authors: Kapustin Alexander N. , Davies John D. , Reynolds Joanne L. , McNair Rosamund , Jones Gregory T. , Sidibe Anissa , Schurgers Leon J. , Skepper Jeremy N. , Proudfoot Diane , Mayr Manuel , Shanahan Catherine M.
Published: May 2011

Rationale:

Matrix vesicles (MVs) are specialized structures that initiate mineral nucleation during physiological skeletogenesis. Similar vesicular structures are deposited at sites of pathological vascular calcification, and studies in vitro have shown that elevated levels of extracellular calcium (Ca) can induce mineralization of vascular smooth muscle cell (VSMC)–derived MVs.

Objectives:

To determine the mechanisms that promote mineralization of VSMC-MVs in response to calcium stress.

Methods and Results:

Transmission electron microscopy showed that both nonmineralized and mineralized MVs were abundantly deposited in the extracellular matrix at sites of calcification. Using cultured human VSMCs, we showed that MV mineralization is calcium dependent and can be inhibited by BAPTA-AM. MVs released by VSMCs in response to extracellular calcium lacked the key mineralization inhibitor matrix Gla protein and showed enhanced matrix metalloproteinase-2 activity. Proteomics revealed that VSMC-MVs share similarities with chondrocyte-derived MVs, including enrichment of the calcium-binding proteins annexins (Anx) A2, A5, and A6. Biotin cross-linking and flow cytometry demonstrated that in response to calcium, AnxA6 shuttled to the plasma membrane and was selectively enriched in MVs. AnxA6 was also abundant at sites of vascular calcification in vivo, and small interfering RNA depletion of AnxA6 reduced VSMC mineralization. Flow cytometry showed that in addition to AnxA6, calcium induced phosphatidylserine exposure on the MV surface, thus providing hydroxyapatite nucleation sites.

Conclusions:

In contrast to the coordinated signaling response observed in chondrocyte MVs, mineralization of VSMC-MVs is a pathological response to disturbed intracellular calcium homeostasis that leads to inhibitor depletion and the formation of AnxA6/phosphatidylserine nucleation complexes.
Reprogramming of Skeletal Myoblasts for Induction of Pluripotency for Tumor-Free Cardiomyogenesis in the Infarcted Heart
Authors: Ahmed Rafeeq P.H. , Haider Husnain K. , Buccini Stephanie , Li Longhu , Jiang Shujia , Ashraf Muhammad
Published: May 2011

Rationale:

Skeletal myoblasts (SMs) with inherent myogenic properties are better candidates for reprogramming to pluripotency.

Objective:

To reprogram SMs to pluripotency and show that reprogrammed SMs (SiPS) express embryonic gene and microRNA profiles and that transplantation of predifferentiated cardiac progenitors reduce tumor formation.

Methods and Results:

The pMXs vector containing mouse cDNAs for Yamanaka's quartet of stemness factors were used for transduction of SMs purified from male Oct4-GFP
+
transgenic mouse. Three weeks later, GFP
+
colonies of SiPS were isolated and propagated in vitro. SiPS were positive for alkaline phosphatase, expressed SSEA1, and displayed a panel of embryonic stem (ES) cell–specific pluripotency markers. Embryoid body formation yielded beating cardiomyocyte-like cells, which expressed early and late cardiac-specific markers. SiPS also had an microRNA profile that was altered during their cardiomyogenic differentiation. Noticeable abrogation of let-7 family and significant up-regulation of miR-200a-c was observed in SiPS and SiPS-derived cardiomyocytes, respectively. In vivo studies in an experimental model of acute myocardial infarction showed extensive survival of SiPS and SiPS-derived cardiomyocytes in mouse heart after transplantation. Our results from 4-week studies in DMEM without cells (group 1), SMs (group-2), SiPS (group-3), and SiPS-derived cardiomyocytes (group 4) showed extensive myogenic integration of the transplanted cells in group 4 with attenuated infarct size and improved cardiac function without tumorgenesis.

Conclusions:

Successful reprogramming was achieved in SMs with ES cell-like microRNA profile. Given the tumorgenic nature of SiPS, their predifferentiation into cardiomyocytes would be important for tumor-free cardiogenesis in the heart.
Neonatal Mouse–Derived Engineered Cardiac Tissue
Authors: de Lange W.J. , Hegge L.F. , Grimes A.C. , Tong C.W. , Brost T.M. , Moss R.L. , Ralphe J.C.
Published: May 2011

Rationale:

Cardiomyocytes cultured in a mechanically active 3-dimensional configuration can be used for studies that correlate contractile performance to cellular physiology. Current engineered cardiac tissue (ECT) models use cells derived from either rat or chick hearts. Development of a murine ECT would provide access to many existing models of cardiac disease and open the possibility of performing targeted genetic manipulation with the ability to directly assess contractile and molecular variables.

Objective:

To generate, characterize, and validate mouse ECT with a physiologically relevant model of hypertrophic cardiomyopathy.

Methods and Results:

We generated mechanically integrated ECT using isolated neonatal mouse cardiac cells derived from both wild-type and myosin-binding protein C (cMyBP-C)–null mouse hearts. The murine ECTs produced consistent contractile forces that followed the Frank-Starling law and accepted physiological pacing. cMyBP-C–null ECTs showed characteristic acceleration of contraction kinetics. Adenovirus-mediated expression of human cMyBP-C in murine cMyBP-C–null ECT restored contractile properties to levels indistinguishable from those of wild-type ECT. Importantly, the cardiomyocytes used to construct the cMyBP-C
−/−
ECT had yet to undergo the significant hypertrophic remodeling that occurs in vivo. Thus, this murine ECT model reveals a contractile phenotype that is specific to the genetic mutation rather than to secondary remodeling events.

Conclusions:

Data presented here show mouse ECT to be an efficient and cost-effective platform to study the primary effects of genetic manipulation on cardiac contractile function. This model provides a previously unavailable tool to study specific sarcomeric protein mutations in an intact mammalian muscle system.
A Model of Canine Purkinje Cell Electrophysiology and Ca
2+
Cycling
Authors: Li Pan , Rudy Yoram
Published: May 2011

Purkinje cells (Pcell) are characterized by different electrophysiological properties and Ca
2+
cycling processes than ventricular myocytes (Vcell) and are frequently involved in ventricular arrhythmias. Yet, the mechanistic basis for their arrhythmic vulnerability is not completely understood. The objectives were to: (1) characterize Pcell electrophysiology, Ca
2+
cycling, and their rate dependence; (2) investigate mechanisms underlying Pcell arrhythmogenicity; and compare Pcell and Vcell electrophysiology, Ca
2+
cycling, and arrhythmic properties. We developed a new mathematical model of Pcell. The Ca
2+
subsystem includes spatial organization and receptors distribution unique to Pcell. Results were: (1) in Pcell and Vcell, Na
+
accumulation via its augmentation of repolarizing I
NaK
dominates action potential duration adaptation and, in Pcell, I
NaL
contributes additional action potential duration shortening at short cycle length; (2) steep Pcell restitution is attributable to slow recovery of I
NaL
; (3) biphasic Ca
2+
transients of Pcell reflect the delay between Ca
2+
release from junctional sarcoplasmic reticulum and corbular sarcoplasmic reticulum; (4) Pcell Ca
2+
alternans, unlike Vcell, can develop without inducing action potential alternans; (5) Pcell action potential alternans develops at a shorter cycle length than Vcell, with increased subcellular heterogeneity of Ca
2+
cycling attributable to refractoriness of Ca
2+
release from corbular sarcoplasmic reticulum and junctional sarcoplasmic reticulum; (6) greater Pcell vulnerability to delayed afterdepolarizations is attributable to higher sarcoplasmic reticulum Ca
2+
content and ionic currents that reduce excitation threshold and promote triggered activity; and (7) early after depolarizations generation in Pcell is mostly attributable to reactivation of I
NaL2
, whereas I
CaL
plays this role in Vcell. Steeper rate dependence of action potential and Ca
2+
transients, central peripheral heterogeneity of Ca
2+
cycling, and distinct ion channel profile underlie greater arrhythmic vulnerability of Pcell compared to Vcell.
Rewriting the Captivity Narrative for Contemporary Children: Speare, Bruchac, and the French and Indian War
Authors: Schwebel Sara L.
Published: May 2011

Juxtaposing the French and Indian War stories of Elizabeth George Speare, a mid-twentieth- century Anglo-American children's author, against those of Joseph Bruchac, a twenty-first- century Abenaki children's author, reveals how flexible and powerful captivity narratives have been in shaping arguments about gender, nationhood, citizenship, and land in the postwar United States.
Understanding Initiation Behavior in Chinese Negotiations: An Examination of Distinctions across Three Regional Subcultures
Authors: Volkema Roger J.
Published: May 2011

While many individuals speak of China as a single trading partner, in reality Greater China consists of at least three regional subcultures ‐ the People’s Republic of China, Hong Kong, and Taiwan. These regions differ in terms of their historical development, values, and traditions, which have implications for what to expect in negotiations. This article examines the cultural differences found in these three regions of Greater China, with particular attention to an often overlooked yet critical stage of the negotiation process ‐ the initiation stage. Using data from the GLOBE Study on cultural practices and values, propensity to initiate a negotiation (engage a counterpart, make a request or demand, and optimize that request) is estimated for each regional subculture. The implications of these findings for practitioners and future research are discussed.
Talk of Corruption in Eastern Indonesian Communities: Reactions to Local Government in the Post-Suharto Reform Era
Authors: Erb Maribeth
Published: May 2011

Eastern Indonesia, most notably Nusa Tenggara Timor province, has been frequently referred to as the poorest region in Indonesia and claims have recently been made that it is the most corrupt as well. The spread of corruption in the post-Suharto period, with the introduction of regional autonomy and decentralisation, has often been commented on; but what is corruption? How do people define it? This paper uses an anthropological lens on corruption to suggest that with the spread of ideas of ‘good governance’ and ‘democracy’, one significant way that local communities in NTT province engage with the state and define corruption is as an abuse of power and non-consultation with the populace. This has become particularly acute with the government belief that new investment opportunities in the reform era will become a road to development and prosperity for local communities, who are, however, seen to be unable to provide for themselves or seek their own ways out of poverty. Several regional governments in NTT province have taken advantage of new laws put into effect in the reform era to award mining concessions to domestic and foreign mining companies. A swell of protest has arisen across the province, however, and an increasing critique of poor government and corrupt practices focuses on these mining contracts.
Sawerigading vs. Sharia: Identities and Political Contestation in Decentralised Indonesia
Authors: Robinson Kathryn
Published: May 2011

Islamist groups are attempting to shape Indonesia’s political landscape post-New Order through advocating local regulations based on sharia in districts newly empowered by regional autonomy. In the province of South Sulawesi, which was gripped by a separatist Islamic rebellion in the 1950s and 1960s, the former rebel leader, Kahar Muzakkar, is invoked in a movement to implement sharia-based local regulations. However, the politics of decentralisation are also associated with a resurgence of local cultural identities, which embrace non-Islamic traditions. In Muslim South Sulawesi, these claims have been expressed through the ceremonial re-installation of local traditional rulers and performance of public ceremonies to care for the sacred regalia that legitimate authority, but also through government-funded seminars that explore distinctive Bugis and Makassarese cultural traditions. These claims to power can be understood as a reaction to the taming of cultural difference by the Suharto regime, but they also represent vehicles for local elites to assume power. Based on an analysis of one of the district cultural seminars and accompanying cultural festival, this paper examines the manner in which cultural traditions are strategically mobilised in South Sulawesi, in a rival movement to the Islamist claims to implement sharia.
Translocal Networks and Globalisation within Indonesia: Exploring the Hadhrami Diaspora from the Archipelago’s North-East
Authors: Slama Martin
Published: May 2011

This article contributes to the study of diasporic aspects of today’s Eastern Indonesia by emphasizing its long history of translocal connections, as well as the multiplicity and acceleration of contemporary flows. It focuses on the Hadhrami diaspora, i.e., Indonesians of Arabic descent whose ancestors migrated from the Hadhramaut (located in today’s Republic of Yemen) to almost all parts of the archipelago. Today, Hadhramis in north-eastern Indonesia, the region this article concentrates on, form distinct communities and maintain translocal networks. There already in colonial times, Hadhramis institutionalised their networks through an Islamic organisation called Al-Khairaat. The article investigates how the translocal networks in north-eastern Indonesia have developed, especially during the New Order and afterwards, when Hadhramis from the region increasingly incorporated Java, especially Jakarta, into their networks. Reconsidering post-Suharto transformations that are part of a globalisation within Indonesia, the article concludes that their networks connect now the north-eastern periphery and Indonesia’s centre more intensively than ever. And, as a consequence of this expansion, Hadhramis from north-eastern Indonesia were able to enter national elite circles in Jakarta.
Jewish Novellas in a Greek and Roman Age: Fiction and Identity
Authors: Wills Lawrence M.
Published: May 2011

Although Jewish novellas (Esther, Daniel, Tobit, Judith, and Joseph and Aseneth) have received more attention recently as a distinct genre within ancient Jewish literature, their relation to Greek and Roman novels is still debated. This article argues that, although some of the Jewish novellas arise earlier, they should be considered part of the same broad category of novelistic literature. The rich research on the cultural context of Greek and Roman novels applies to the Jewish as well. But a further question is also explored: if the Jewish texts were originally considered fictional, how did they come to be considered biblical and historical? Two suggestions are proposed: the protagonists of the narratives first came to be revered as heroes of the faith aside from the texts, and the rise of “biblical history” required the use of Esther and Daniel to fill in the gaps in the chronology.
Stringency in Qumran?
Authors: Heger Paul
Published: May 2011

Some scholars allege a stringent ideology behind the severe Qumranic halakhot in contrast to putative leniencies in parallel rabbinic rulings. Vered Noam contends that the rabbinic propensity for leniency and innovativeness, versus the opposite Qumranic approach are the source of their disputes. Thus, Qumranic strictness is not “objective” but relative to the lenient rabbinic law. This paper critically scrutinizes Noam’s thesis, and will posit that the halakhot of Qumran are founded on a literal adherence to Scripture versus rabbinic pragmatism. The study discusses Noam’s cited examples, offering contrasting explanations. It deliberates about the difference between disputes concerning interpretations and physical facts, the rationales behind the various classifications, and the relationship between the degrees of holiness and the degrees of impurity.
Conversos and Novamente Convertidos: Law, Religion, and Identity in the Portuguese Kingdom and Empire
Authors: Xavier Ângela Barreto
Published: May 2011

In this essay, I intend to demonstrate that the foundations of the attitudes of the Portuguese crown and its agents in relation to the native populations living in the imperial territories were mainly inspired by the relations in Portugal with Jews, and later, Conversos. Besides the strong influence of classical inspiration and a rereading of medieval travelers on the construction of the imperial territories, the ways of thinking about, identifying, and governing internal others served as a reference guide to interpret and model the social situations that emerged in the Portuguese imperial territories. Indeed, the establishment of analogies between the populations of the kingdom and imperial populations had a great practical impact, and molded a good part of the political solutions that were delineated initially as extra territorium, particularly those in the territory of Goa.
Learning by Example: Exempla in Jerome’s Translations and Revisions of Biblical Books
Authors: Skemp Vincent T.M.
Published: May 2011

Jerome’s interest in provinding Exempla for his readers in his letters and the Vitae Patrum extended to his translations and revisions of canonical Books—Ruth 4,11; Deut 29,22(23), Esth 1,18; Zech 13,4—and to a book he considered non-canonical (Tob 2,12). Jerome related the noun exemplum to a variety of lexemes in the semantic field of obiecta aversionis. His views regarding the regulation of female behavior played a role in the plus in Esth 1,18. He viewed Ruth to some extent as a positive exemplum for his female audience, although he likely had conflicted feelings about the fact that the widow remarried. Although the lexeme does not occur in the Book of Judith, his predilections for Judith as exemplum of the chaste and virtuous widow was the impetus for the pluses in Jdt 16,26 and 10,4.
Missional Churches in Secular Societies: Theology Consults Sociology
Authors: Hughson, S.J. Thomas
Published: May 2011

The church is missionary by nature. But what about public church mission in secular societies? Furious religion mobilizing against rebarbative secularity? Withdrawal to seek exemplary perfection? To the contrary, theologically principled consultation with the sociology of J. Casanova on deprivatized religion leads to public witness in modern societies. Public theology can interpret deprivatized religion as an expression of prophetic and kingly elements in church mission. However, sociology leaves the priestly element as if private. What might ecclesiology, missiology, and public theology say about a public aspect of the priestly element in the church's witness in modern societies?