bluuebunny/crossref_metadata_2025 huggingface dataset:
Caveolin-1 regulates Mcl-1 stability and anoikis in lung carcinoma cells
Authors: Chunhacha Preedakorn , Pongrakhananon Varisa , Rojanasakul Yon , Chanvorachote Pithi
Published: January 2012

Both caveolin-1 (Cav-1) and Mcl-1 have been implicated in the regulation of cancer cell anoikis, but their relationship and underlying mechanisms of regulation are not known. The present study demonstrated for the first time that Cav-1 regulates Mcl-1 through protein-protein interaction and inhibits its downregulation during cell anoikis in human lung cancer cells. Immunoprecipitation and immunocytochemistry studies showed that Cav-1 interacted with Mcl-1 and prevented it from degradation via the ubiquitin-proteasome pathway. Mcl-1 and Mcl-1-Cav-1 complex were highly elevated in Cav-1-overexpressing cells but were greatly reduced in Cav-1 knockdown cells. Consistent with this finding, we found that Mcl-1 ubiquitination was significantly attenuated by Cav-1 overexpression but increased by Cav-1 knockdown. Together, our results indicate a novel role of Cav-1 in anoikis regulation through Mcl-1 interaction and stabilization, which provides a new insight to the pathogenesis of metastatic lung cancer and its potential treatment.
The Na-K-ATPase α1β1 heterodimer as a cell adhesion molecule in epithelia
Authors: Vagin Olga , Dada Laura A. , Tokhtaeva Elmira , Sachs George
Published: January 2012

The ion gradients generated by the Na-K-ATPase play a critical role in epithelia by driving transepithelial transport of various solutes. The efficiency of this Na-K-ATPase-driven vectorial transport depends on the integrity of epithelial junctions that maintain polar distribution of membrane transporters, including the basolateral sodium pump, and restrict paracellular diffusion of solutes. The review summarizes the data showing that, in addition to pumping ions, the Na-K-ATPase located at the sites of cell-cell junction acts as a cell adhesion molecule by interacting with the Na-K-ATPase of the adjacent cell in the intercellular space accompanied by anchoring to the cytoskeleton in the cytoplasm. The review also discusses the experimental evidence on the importance of a specific amino acid region in the extracellular domain of the Na-K-ATPase β1 subunit for the Na-K-ATPase trans-dimerization and intercellular adhesion. Furthermore, a possible role of N-glycans linked to the Na-K-ATPase β1 subunit in regulation of epithelial junctions by modulating β1-β1 interactions is discussed.
Exogenous GLP-2 and IGF-I induce a differential intestinal response in IGF binding protein-3 and -5 double knockout mice
Authors: Murali Sangita G. , Brinkman Adam S. , Solverson Patrick , Pun Wing , Pintar John E. , Ney Denise M.
Published: January 2012

Glucagon-like peptide-2 (GLP-2) action is dependent on intestinal expression of IGF-I, and IGF-I action is modulated by IGF binding proteins (IGFBP). Our objective was to evaluate whether the intestinal response to GLP-2 or IGF-I is dependent on expression of IGFBP-3 and -5. Male, adult mice in six treatment groups, three wild-type (WT) and three double IGFBP-3/-5 knockout (KO), received twice daily intraperitoneal injections of GLP-2 (0.5 μg/g body wt), IGF-I (4 μg/g body wt), or PBS (vehicle) for 7 days. IGFBP-3/-5 KO mice showed a phenotype of lower plasma IGF-I concentration, but greater body weight and relative mass of visceral organs, compared with WT mice ( P < 0.001). WT mice showed jejunal growth with either IGF-I or GLP-2 treatment. In KO mice, IGF-I did not stimulate jejunal growth, crypt mitosis, sucrase activity, and IGF-I receptor (IGF-IR) expression, suggesting that the intestinotrophic actions of IGF-I are dependent on expression of IGFBP-3 and -5. In KO mice, GLP-2 induced significant increases in jejunal mucosal cellularity, crypt mitosis, villus height, and crypt depth that was associated with increased expression of the ErbB ligand epiregulin and decreased expression of IGF-I and IGF-IR. This suggests that in KO mice, GLP-2 action in jejunal mucosa is independent of the IGF-I system and linked with ErbB ligands. In summary, the intestinotrophic actions of IGF-I, but not GLP-2, in mucosa are dependent on IGFBP-3 and -5. These findings support the role of multiple downstream mediators for the mucosal growth induced by GLP-2.
Early depression of Ankrd2 and Csrp3 mRNAs in the polyribosomal and whole tissue fractions in skeletal muscle with decreased voluntary running
Authors: Roberts Michael D. , Childs Thomas E. , Brown Jacob D. , Davis J. Wade , Booth Frank W.
Published: January 2012

The wheel-lock (WL) model for depressed ambulatory activity in rats has shown metabolic maladies ensuing within 53–173 h after WL begins. We sought to determine if WL beginning after 21–23 days of voluntary running in growing female Wistar rats affected the mRNA profile in the polyribosomal fraction from plantaris muscle shortly following WL. In experiment 1, WL occurred at 0200 and muscles were harvested at 0700 daily at 5 h (WL5h, n = 4), 29 h (WL29h, n = 4), or 53 h (WL53h, n = 4) after WL. Affymetrix Rat Gene 1.0 ST Arrays were used to test the initial question as to whether WL affects mRNA occupancy on skeletal muscle polyribosomes. Using a false discovery rate of 15%, no changes in mRNAs in the polyribosomal fraction were observed at WL29h and eight mRNAs (of over 8,200 identified targets) were altered at WL53h compared with WL5h. Interestingly, two of the six downregulated genes included ankyrin repeat domain 2 (Ankrd2) and cysteine-rich protein 3/muscle LIM protein (Csrp3), both of which encode mechanical stretch sensors and RT-PCR verified their WL-induced decline. In experiment 2, whole muscle mRNA and protein levels were analyzed for Ankrd2 and Csrp3 from the muscles of WL5h (4 original samples + 2 new), WL29h (4 original), WL53h (4 original + 2 new), as well as WL173 h ( n = 6 new) and animals that never ran (SED, 4–5 new). Relative to WL5h controls, whole tissue Ankrd2 and Csrp3 mRNAs were lower ( P < 0.05) at WL53h, WL173h, and SED; Ankrd2 protein tended to decrease at WL53h ( P = 0.054) and Csrp3 protein was less in WL173h and SED rats ( P < 0.05). In summary, unique early declines in Ankrd2 and Csrp3 mRNAs were identified with removal of voluntary running, which was subsequently followed by declines in Csrp3 protein levels during longer periods of wheel lock.
Evolutionary Perspectives on Myrmecophily in Ladybirds
Authors: Vantaux Amélie , Roux Olivier , Magro Alexandra , Orivel Jérôme
Published: January 2012

Myrmecophiles are species that usually have developed specialized traits to cope with the aggressiveness of ants enabling them to live in their vicinity. Many coccinellid species are predators of Hemiptera; the latter is also often protected by ants. Therefore these ladybirds frequently interact with ants, and some species have become myrmecophilous. In this paper, we aim to provide an overview of the evolution of myrmecophilous traits in ladybirds. We then discuss the costs and benefits of myrmecophily and the dietary shift to myrmecophagy observed in a few species.
BMK1 Kinase Suppresses Epithelial–Mesenchymal Transition through the Akt/GSK3β Signaling Pathway
Authors: Chen Runqiang , Yang Qingkai , Lee Jiing-Dwan
Published: January 2012

Epithelial–mesenchymal transition (EMT) plays a crucial role in the development of cancer metastasis. The mitogen-activated protein (MAP) kinases extracellular signal–regulated kinase, c-jun-NH2-kinase, and p38 have been implicated in promoting EMT, but a role for the MAP kinase BMK1 has not been studied. Here, we report that BMK1 signaling suppresses EMT. BMK1 elevation augmented E-cadherin–mediated cell–cell adhesion, downregulated mesenchymal markers, and decreased cell motility. Conversely, BMK1 silencing attenuated E-cadherin–mediated cell–cell adhesion, upregulated mesenchymal markers, and stimulated cell motility. BMK1 depletion dramatically increased the accumulation of endogenous Snail in the nuclear compartment. Snail accumulation was mediated by Akt/GSK3β signaling, which was activated by a modulation in the expression of the mTOR inhibitor DEPTOR. In support of these observations, BMK1 depletion promoted metastasis in vivo. Together, our findings reveal a novel mechanism of EMT control via mTOR/Akt inhibition that suppresses cancer metastasis. Cancer Res; 72(6); 1579–87. ©2012 AACR.
Ultrasound Increases Nanoparticle Delivery by Reducing Intratumoral Pressure and Increasing Transport in Epithelial and Epithelial–Mesenchymal Transition Tumors
Authors: Watson Katherine D. , Lai Chun-Yen , Qin Shengping , Kruse Dustin E. , Lin Yueh-Chen , Seo Jai Woong , Cardiff Robert D. , Mahakian Lisa M. , Beegle Julie , Ingham Elizabeth S. , Curry Fitz-Roy , Reed Rolf K. , Ferrara Katherine W.
Published: January 2012

Acquisition of the epithelial–mesenchymal transition (EMT) tumor phenotype is associated with impaired chemotherapeutic delivery and a poor prognosis. In this study, we investigated the application of therapeutic ultrasound methods available in the clinic to increase nanotherapeutic particle accumulation in epithelial and EMT tumors by labeling particles with a positron emission tomography tracer. Epithelial tumors were highly vascularized with tight cell–cell junctions, compared with EMT tumors where cells displayed an irregular, elongated shape with loosened cell–cell adhesions and a reduction in E-cadherin and cytokeratins 8/18 and 19. Without ultrasound, the accumulation of liposomal nanoparticles administered to tumors in vivo was approximately 1.5 times greater in epithelial tumors than EMT tumors. When ultrasound was applied, both nanoaccumulation and apparent tumor permeability were increased in both settings. Notably, ultrasound effects differed with thermal and mechanical indices, such that increasing the thermal ultrasound dose increased nanoaccumulation in EMT tumors. Taken together, our results illustrate how ultrasound can be used to enhance nanoparticle accumulation in tumors by reducing their intratumoral pressure and increasing their vascular permeability. Cancer Res; 72(6); 1485–93. ©2012 AACR.
Direct Therapeutic Applications of Calcium Electroporation to Effectively Induce Tumor Necrosis
Authors: Frandsen Stine Krog , Gissel Hanne , Hojman Pernille , Tramm Trine , Eriksen Jens , Gehl Julie
Published: January 2012

Electroporation of cells with short, high-voltage pulses causes a transient permeabilization of cell membranes that permits passage of otherwise nonpermeating ions and molecules. In this study, we illustrate how electroporation with isotonic calcium can achieve highly effective cancer cell kill in vivo. Calcium electroporation elicited dramatic antitumor responses in which 89% of treated tumors were eliminated. Histologic analyses indicated complete tumor necrosis. Mechanistically, calcium electroporation caused acute ATP depletion likely due to a combination of increased cellular use of ATP, decreased production of ATP due to effects on the mitochondria, as well as loss of ATP through the permeabilized cell membrane. Taken together, our findings offer a preclinical proof of concept for the use of electroporation to load cancer cells with calcium as an efficient anticancer treatment. Electroporation equipment is already used clinically to enhance the delivery of chemotherapy to superficial tumors, with trials on internal tumors in progress, enabling the introduction of calcium electroporation to clinical use. Moreover, the safety profile, availability, and low cost of calcium facilitate access to this technology for many cancer patients in developed and developing countries. Cancer Res; 72(6); 1336–41. ©2012 AACR.
Ribosome Biogenesis and Control of Cell Proliferation: p53 Is Not Alone
Authors: Donati Giulio , Montanaro Lorenzo , Derenzini Massimo
Published: January 2012

Cell growth is a prerequisite for cell proliferation, and ribosome biogenesis is a limiting factor for cell growth. In mammalian cells, the tumor suppressor p53 has been shown to induce cell-cycle arrest in response to impaired ribosome biogenesis. Recently, p53-independent mechanisms of cell-cycle arrest in response to alterations of ribosome biogenesis have been described. These findings provide a rational basis for the use of drugs that specifically impact ribosome biogenesis for the treatment of cancers lacking active p53 and extend the scenario of mechanisms involved in the relationship between cell growth and cell proliferation. Cancer Res; 72(7); 1602–7. ©2012 AACR.
Eribulin Mesylate for the Treatment of Patients with Refractory Metastatic Breast Cancer: Use of a “Physician's Choice” Control Arm in a Randomized Approval Trial
Authors: Donoghue Martha , Lemery Steven J. , Yuan Weishi , He Kun , Sridhara Rajeshwari , Shord Stacy , Zhao Hong , Marathe Anshu , Kotch Lori , Jee Josephine , Wang Ying , Zhou Liang , Adams William M. , Jarral Vaishali , Pilaro Anne , Lostritto Richard , Gootenberg Joseph E. , Keegan Patricia , Pazdur Richard
Published: January 2012

This work describes the considerations that led to the approval by the U.S. Food and Drug Administration (FDA), on November 15, 2010, of eribulin mesylate (Halaven; Eisai, Inc.) for the treatment of patients with refractory metastatic breast cancer. The FDA review focused primarily on the results of a single randomized, open-label, multicenter trial of 762 patients with refractory locally advanced or metastatic breast cancer. The patients were randomized to receive eribulin or any single-agent treatment of the physician's choice, selected prior to randomization. The FDA's approval of eribulin mesylate was based on demonstration of a statistically significant prolongation of overall survival (OS) in patients who had been randomized to receive eribulin. The median OS was 13.1 months in the eribulin arm compared with 10.6 months in the control arm [HR 0.81 (95% CI, 0.66–0.99); P = 0.041]. Treatment with eribulin did not show a statistically significant treatment effect [HR 0.87 (95% CI, 0.71–1.05)] on progression-free survival as determined by independent review. This approval highlights the appropriate use of an innovative trial design and shows that improvement in OS is an achievable endpoint in the setting of advanced breast cancer. On the basis of the different conclusions arising from the OS and progression-free survival results, investigators should consider using OS as a primary endpoint in clinical trials for refractory breast cancer. Clin Cancer Res; 18(6); 1496–505. ©2012 AACR.
KRAS and BRAF Mutations Predict Primary Resistance to Imatinib in Gastrointestinal Stromal Tumors
Authors: Miranda Claudia , Nucifora Martina , Molinari Francesca , Conca Elena , Anania Maria Chiara , Bordoni Andrea , Saletti Piercarlo , Mazzucchelli Luca , Pilotti Silvana , Pierotti Marco A. , Tamborini Elena , Greco Angela , Frattini Milo
Published: January 2012

Purpose: Gastrointestinal stromal tumors (GIST) are characterized by gain-of-function mutations in KIT/PDGFRA genes leading to a constitutive receptor activation which is well counteracted by imatinib. However, cases in which imatinib as first-line treatment has no effects are reported (primary resistance). Our purpose is to investigate alterations in downstream effectors, not reported so far in mutated GIST, possibly explaining the primary resistance to targeted treatments.

Experimental Design: Two independent naive GIST cohorts have been analyzed for KIT, PDGFRA, KRAS, and BRAF mutations by direct sequencing. Cell lines expressing a constitutively activated and imatinib-responding KIT, alone or in combination with activated KRAS and BRAF, were produced and treated with imatinib. KIT receptor and its downstream effectors were analyzed by direct Western blotting.

Results: In naive GISTs carrying activating mutations in KIT or PDGFRA a concomitant activating mutation was detected in KRAS (5%) or BRAF (about 2%) genes. In vitro experiments showed that imatinib was able to switch off the mutated receptor KIT but not the downstream signaling triggered by RAS–RAF effectors.

Conclusions: These data suggest the activation of mitogen—activated protein kinase pathway as a possible novel mechanism of primary resistance to imatinib in GISTs and could explain the survival curves obtained from several clinical studies where 2% to 4% of patients with GIST treated with imatinib, despite carrying KIT-sensitive mutations, do not respond to the treatment. Clin Cancer Res; 18(6); 1769–76. ©2012 AACR.
Genome-Wide miRNA Expression Profiling Identifies miR-9-3 and miR-193a as Targets for DNA Methylation in Non–Small Cell Lung Cancers
Authors: Heller Gerwin , Weinzierl Marlene , Noll Christian , Babinsky Valerie , Ziegler Barbara , Altenberger Corinna , Minichsdorfer Christoph , Lang György , Döme Balazs , End-Pfützenreuter Adelheid , Arns Britt-Madeleine , Grin Yuliya , Klepetko Walter , Zielinski Christoph C. , Zöchbauer-Müller Sabine
Published: January 2012

Purpose: The major aim of this study was to investigate the role of DNA methylation (referred to as methylation) on miRNA silencing in non–small cell lung cancers (NSCLC).

Experimental Design: We conducted microarray expression analyses of 856 miRNAs in NSCLC A549 cells before and after treatment with the DNA methyltransferase inhibitor 5-aza-2′-deoxycytidine (Aza-dC) and with a combination of Aza-dC and the histone deacetylase inhibitor trichostatin A. miRNA methylation was determined in 11 NSCLC cell lines and in primary tumors and corresponding nonmalignant lung tissue samples of 101 patients with stage I–III NSCLC.

Results: By comparing microarray data of untreated and drug-treated A549 cells, we identified 33 miRNAs whose expression was upregulated after drug treatment and which are associated with a CpG island. Thirty (91%) of these miRNAs were found to be methylated in at least 1 of 11 NSCLC cell lines analyzed. Moreover, miR-9-3 and miR-193a were found to be tumor specifically methylated in patients with NSCLC. We observed a shorter disease-free survival of patients with miR-9-3 methylated lung squamous cell carcinoma (LSCC) than patients with miR-9-3 unmethylated LSCC by multivariate analysis [HR = 3.8; 95% confidence interval (CI), 1.3–11.2, P = 0.017] and a shorter overall survival of patients with miR-9-3 methylated LSCC than patients with miR-9-3 unmethylated LSCC by univariate analysis (P = 0.013).

Conclusions: Overall, our results suggest that methylation is an important mechanism for inactivation of certain miRNAs in NSCLCs and that miR-9-3 methylation may serve as a prognostic parameter in patients with LSCC. Clin Cancer Res; 18(6); 1619–29. ©2012 AACR.
Liver-Intestine Cadherin Expression Is Associated with Intestinal Differentiation and Carcinogenesis in Intraductal Papillary Mucinous Neoplasm
Authors: Morimatsu Katsuya , Aishima Shinichi , Kayashima Tadashi , Hayashi Akifumi , Nakata Kohei , Oda Yasunori , Taguchi Tomoaki , Tsuneyoshi Masazumi , Tanaka Masao , Oda Yoshinao
Published: January 2012

Objectives: Intraductal papillary mucinous neoplasms (IPMN) are classified into four phenotypes according to the WHO classification. Recently, intestinal-type IPMN has been suggested to grow with a distinct carcinogenetic pathway. Like mucin 2, oligomeric mucus/gel forming (MUC2) and caudal-related homeobox 2 (CDX2), liver-intestine cadherin (LI cadherin) is an intestine-specific marker. We aimed to investigate the roles of LI cadherin expression in IPMN. Methods: We examined LI cadherin expression in 135 cases of IPMN by immunohistochemical staining and the quantitative real-time reverse-transcription polymerase chain reaction. Results: LI cadherin protein and mRNA levels were significantly higher in intestinal-type IPMN than in nonintestinal-type IPMN (protein level, p < 0.001; mRNA level, p = 0.0312). A positive correlation was found between protein and mRNA of LI cadherin (p = 0.0037). The positivity rates of LI cadherin expression were significantly higher in CDX2-positive cases than in CDX2-negative cases (p < 0.001). In 41 intestinal-type IPMNs, LI cadherin-positive rates tended to increase gradually, from IPMN with low-grade dysplasia (IPMN-L) to IPMN with an associated invasive carcinoma (IPMN-IC) [IPMN-L vs. IPMN with high-grade dysplasia (IPMN-H); p = 0.0357, IPMN-L vs. IPMN-IC; p = 0.0230] and positively correlated with the Ki-67 labeling index (p = 0.0408), whereas this tendency was not recognized in nonintestinal-type IPMNs. Conclusions: LI cadherin is associated with an intestinal phenotype and an ‘intestinal pathway’ of carcinogenesis in IPMN.
Expression of the Mammalian Target of Rapamycin Pathway Markers in Lung Adenocarcinoma and Squamous Cell Carcinoma
Authors: Kim Hyun-Soo , Kim Gou Young , Lim Sung-Jig , Kim Youn Wha
Published: January 2012

Objective: We investigated whether the expression of mammalian target of rapamycin (mTOR) pathway components is associated with clinicopathologic characteristics and patient outcome in lung adenocarcinoma (AC) and squamous cell carcinoma (SCC). Methods: We used immunohistochemistry to evaluate the expression of phosphorylated Akt (pAkt), mTOR, p70 ribosomal protein S6 kinase (p70S6K) and phosphatase and tensin homolog deleted on chromosome 10 (PTEN) in 91 cases of AC and 154 cases of SCC. Results: pAkt expression was positively correlated with the expression of mTOR (p < 0.001) and p70S6K (p < 0.001), and mTOR expression was positively correlated with p70S6K expression (p < 0.001). PTEN expression was inversely correlated with the expression of pAkt (p = 0.001), mTOR (p < 0.001) and p70S6K (p = 0.012). In addition, loss of PTEN expression, observed in 37.4% (34/91) of AC patients, was significantly associated with a higher histologic grade (p = 0.013), pathologic T stage (p = 0.016) and N stage (p < 0.001) and advanced TNM stage (p = 0.001), as well as a shorter overall survival of AC patients (p = 0.015). Conclusion: The high prevalence of PTEN loss and its association with aggressive tumor behavior and poor patient outcome in AC suggest that loss of PTEN expression is involved in AC progression and serves as a prognostic marker for patients with AC.
Hypoxia-Ischemia Upregulates TRAIL and TRAIL Receptors in the Immature Rat Brain
Authors: Huang Zhiheng , Song Lili , Wang Congmin , Liu Jiang-Qin , Chen Chao
Published: January 2012

The immature brain is susceptible to inflammatory injury induced by hypoxia-ischemia (HI) or infection, which causes serious neurodevelopmental disabilities in the survivors of preterm births. Recently, tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) and its receptors (death receptor DR4/5 and decoy receptor DcR1/2) were reported to mediate various neuroinflammatory responses. However, little information is available regarding the role of TRAIL and its receptors in the immature brain after HI. The purpose of this study was to evaluate the expression of TRAIL and its receptors in the immature brain after HI and relate this expression to neurological function. We performed right common carotid artery ligation followed by hypoxia (6% O2, 37°C) for 2.5 h to induce HI in postnatal day 3 rats. The distribution of TRAIL and its receptors, caspase-3 and CD68-labeled microglia/macrophages was evaluated 24 h after HI by immunostaining. The protein and mRNA expression of TRAIL and DR5 was measured by Western blot and real-time PCR, respectively. Delayed neuronal loss was evaluated by NeuN and Nissl staining 7 days after HI. Furthermore, neurological deficits were evaluated by a righting reflex test, time of eye opening and T-maze test. The expression of TRAIL, DR5 and DcR1/2 receptors and caspase-3 was more pronounced in the ipsilateral hemisphere compared with the contralateral part and the control group 24 h after HI. DR5/active caspase-3 double-positive cells were observed at 24 h after HI in the ipsilateral hemisphere but not in the contralateral hemisphere. The TRAIL and CD68 double-labeled cells were more pronounced in the ipsilateral cortical regions compared with the corresponding regions of the contralateral part. HI also resulted in a significant increase in TRAIL and DR5 protein and mRNA expression at 24 h, which corresponded to neuronal cell loss 7 days after HI. Furthermore, the HI group displayed impaired neurobehavioral development compared with the control group (p < 0.05). Altogether our results show that the TNF-α superfamily ligand TRAIL is induced on CD68+ microglia/macrophages after perinatal HI and that one of its receptors, DR5, is induced on neocortical neurons and glial cells. That many DR5+ cells were also caspase-3+ strongly supports the conclusion that these signaling molecules are involved in the delayed loss of neurons in the neocortex and in the neurobehavioral deficits that are often seen after perinatal HI.
Association of Mild to Moderate Kidney Dysfunction with Coronary Artery Calcification in Patients with Suspected Coronary Artery Disease
Authors: Cao Xu-fen , Yan Li-qiu , Han Li-xian , Li Dong-mei , Zhao Rong-cheng , Zheng Ye , Guo Nan , Han Jia , Yu Jing
Published: January 2012

Objectives: To report the association between kidney dysfunction and coronary artery calcification (CAC) score (CACS) among patients with clinically suspected coronary artery disease (CAD). Methods: We prospectively included 1,572 consecutive patients with clinically suspected CAD who underwent ECG-gated cardiac CT scans using 64-multidetector row computed tomography. CACS was quantified using a previously described method. Renal function was assessed by the estimated glomerular filtration rate (eGFR). Ordinal logistic regression and Pearson correlation were used to analyze the association between eGFR and CACS. Results: Patients with higher CACS were older, more had a history of hypertension, diabetes and tobacco use, and they were more likely to have an atherogenic lipid profile, higher systolic blood pressure, diastolic blood pressure, hemoglobin A1c, body mass index and C-reactive protein (CRP) and lower eGFR when compared with those patients without CAC or with lower CACS. The unadjusted correlation coefficient of eGFR and CACS was –0.259 (p < 0.001). This remained significant after adjustment for age, gender, hypertension, diabetes, hyperlipidemia, tobacco use and CRP (R = –0.156, p < 0.001). Ordinal logistic regression analysis showed that age, hypertension, diabetes, CRP and eGFR exerted independent influences on CACS. Conclusions: Kidney dysfunction was an independent predictor of CACS in patients with clinically suspected CAD. Further prospective multicenter studies are needed to confirm this finding.
Pathobiological Implications of MUC16/CA125 Expression in Intrahepatic Cholangiocarcinoma-Mass Forming Type
Authors: Higashi Michiyo , Yamada Norishige , Yokoyama Seiya , Kitamoto Sho , Tabata Kazuhiro , Koriyama Chihaya , Batra Surinder K. , Yonezawa Suguru
Published: January 2012

Objectives: MUC16 carries the peptide epitope CA125, which is well known as a marker of ovarian cancer. High serum levels of MUC16 (CA125) have been reported not only in patients with ovarian cancer but also in patients with liver diseases. We evaluated the expression of MUC16 in intrahepatic cholangiocarcinoma-mass forming type (ICC-MF) tissues. Methods: We examined the expression of MUC16 by immunohistochemical analyses using the monoclonal antibody M11 in ICC-MF tissues from 63 patients. To compare the prevalence of each mucin expression by clinicopathological features, appropriate statistical analysis was performed. Results: MUC16 was detected in 48% of samples (30/63). After adjusting for the effects of other prognostic factors, multivariate survival analysis revealed that MUC16 expression is a significant independent factor of poor prognosis (p = 0.005). Conclusion: The current results indicate that MUC16 expression is a prognostic factor of poor survival in ICC-MF.
Granulocyte Colony-Stimulating Factor Reduces Hyperoxia-Induced Alveolarization Inhibition by Increasing Angiogenic Factors
Authors: Lee Hyun Ju , Choi Chang Won , Kim Ee-Kyung , Kim Han-Suk , Kim Beyong Il , Choi Jung-Hwan
Published: January 2012

Background: Granulocyte colony-stimulating factor (G-CSF) is known to mobilize endothelial progenitor cells (EPCs) from bone marrow. EPCs reportedly promote neovascularization and participate in the repair of lung structure in adult animals. Objective: We tested the hypothesis that G-CSF contributes to alveolar growth by increasing the production of angiogenic growth factor in the lungs of hyperoxia-exposed neonatal mice. Methods: Neonatal mice were exposed to hyperoxia (80%) or room air (RA) for 7 days and treated with G-CSF (50 µg/kg/day) or vehicle for 5 days. Blood was subjected to flow cytometry to gate for CD45dim/–/Sca-1+/CD133+/vascular endothelial growth factor (VEGF) receptor-2 (VEGFR2) to define the EPC population at day 7. Results: The percentage of EPCs in the peripheral blood and VEGF and VEGFR2 levels in the lungs of neonatal mice exposed to hyperoxia were significantly reduced compared to those of mice kept in RA. G-CSF significantly increased EPCs in the peripheral blood, and VEGF and VEGFR2 levels in the lungs of both mice exposed to hyperoxia and mice kept in RA. G-CSF restored alveolarization inhibited by hyperoxia without altering normal alveolarization under RA. Conclusion: G-CSF restored alveolarization inhibited by hyperoxia in the developing lungs and this alveolarization-enhancing effect of G-CSF is associated with mobilization of EPCs and upregulation of VEGF signaling.
Surgical Management of Extremely Low Birth Weight Infants with Neonatal Bowel Perforation: A Single-Center Experience and a Review of the Literature
Authors: Eicher Carmen , Seitz Guido , Bevot Andrea , Moll Monika , Goelz Rangmar , Arand Joerg , Poets Christian , Fuchs Joerg
Published: January 2012

Background: Necrotizing enterocolitis (NEC) and focal intestinal perforation (FIP) are major causes of morbidity in infants with extremely low birth weight (ELBW). Objective: To evaluate the surgical procedures applied, and the survival and long-term outcome of ELBW infants with NEC and FIP in a single-center study. Methods: Inborn and outborn ELBW infants (<1000 g) with NEC and FIP were analyzed retrospectively from 2002 to 2007. Data collected include surgical procedures, survival as well as complications, length of partial parenteral nutrition and hospital stay. The short-term and long-term outcomes after 2–7 years were assessed and compared with a matched control group. Results: Out of 280 ELBW infants, 28 underwent surgery, 19 because of FIP and 9 for NEC. Fourteen infants in the FIP group were treated with primary laparotomy and 5 with peritoneal drainage (PD). In the NEC group, only 1 infant was treated with PD. PD was used for unstable patients and was always followed by secondary laparotomy after stabilization. Five of 28 (18%) surgically treated ELBW infants and 4 (14%) matched controls died. The following complications occurred in the surgical group: complete (n = 1) or minor wound dehiscence (n = 4), stoma prolapse (n = 5), parastomal hernia (n = 2), stoma fistula (n = 1), and wound infection (n = 2). Dependency on parenteral nutrition was significantly shorter in infants with FIP, while there were no differences in time to stoma closure and length of hospital stay between those with FIP and those with NEC. Eleven of 23 (47.8%) surviving patients with FIP or NEC showed developmental delay, compared with 9 of 24 (37.5%) in the controls. Conclusions: The management of EBLW infants with NEC and FIP remains challenging. Our treatment approach was associated with low mortality. Developmental delay seems to be caused by extreme prematurity rather than NEC- or FIP-related bowel perforation.
Pericardial Effusion in Renal Disease: To Tap or Not to Tap
Authors: Krantz Mori J. , Byrd James Brian
Published: January 2012

The natural history of pericardial effusions attributable to renal disease is variable. Although aggressive hemodialysis may lead to the resolution of some effusions, some reports suggest that prompt drainage is optimal. We describe a case of a 49-year-old woman who presented with end-stage renal disease and a large pericardial effusion. Although she was hypertensive on presentation and had no pulsus paradoxus, transthoracic echocardiography revealed stigmata of cardiac tamponade, including right atrial and ventricular collapse, as well as a plethoric inferior vena cava. Because of the lack of certain clinical signs of tamponade and due to concern about excess bleeding risk in the setting of uremia, the effusion was initially managed with serial dialysis rather than pericardiocentesis. The effusion did not decrease in size despite 1 week of hemodialysis, and the patient developed acute dyspnea, relative tachycardia and hypotension after an increase in the blood flow rate during hemodialysis, all of which resolved with a decrease in the blood flow rate. The onset of dyspnea during a session of dialysis as a symptom of tamponade physiology has not been reported previously. We believe that this case supports early pericardiocentesis in patients with any degree of echocardiographic evidence of tamponade. We discuss this in the context of existing literature, which suggests that pericardiocentesis, rather than dialysis, is the preferred management strategy for large uremic pericardial effusions, even in the absence of evidence of clinical signs of pericardial tamponade.
Multiple Sclerosis and Risk of Venous Thromboembolism: A Population-Based Cohort Study
Authors: Christensen Steffen , Farkas Dóra Körmendiné , Pedersen Lars , Miret Montserrat , Christiansen Christian Fynbo , Sørensen Henrik Toft
Published: January 2012

Background: Multiple sclerosis (MS) patients may be at increased risk of venous thromboembolism (VTE), but evidence is limited. Objectives: To examine long-term risk of VTE among MS patients. Patients and Methods: We conducted a population-based cohort study among 17,418 Danish MS patients and 87,090 comparison cohort members from the general population. Data on MS, VTE and comorbidities were obtained from the Danish National Registry of Patients including all admissions to Danish hospitals since 1977. We computed cumulative risks for VTE and adjusted incidence rate ratios (IRRs). Results: A total of 34 (0.2%) MS patients and 36 (0.04%) comparison cohort members had a deep venous thrombosis (DVT) within 1 year following the date of initial MS diagnosis/index date [adjusted IRR = 3.02 (95% CI: 2.14–4.27)]. During this period, 16 (0.09%) MS patients and 26 (0.03%) comparison cohort members had a documented pulmonary embolism (PE) [adjusted IRR = 2.85 (95% CI: 1.72–4.70)]. During the subsequent up to 29 years, 315 (1.9% of MS patients alive at year 1) MS patients had a record of a DVT [adjusted IRR = 2.28 (95% CI: 2.01–2.59)] and 129 (0.8%) had PE [IRR = 1.58 (95% CI: 1.31–1.92]. Conclusion: MS is a risk factor for VTE, but the absolute risk is low.
Stereotactic Biopsy for Brainstem Tumors: Comparison of Transcerebellar with Transfrontal Approach
Authors: Dellaretti Marcos , Reyns Nicolas , Touzet Gustavo , Dubois François , Gusmão Sebastião , Pereira Júlio Leonardo Barbosa , Blond Serge
Published: January 2012

Background: An important aspect of evaluating patients submitted to stereotactic biopsy of the brainstem is the trajectory used. The literature describes two principal approaches: the suboccipital transcerebellar and the transfrontal; however, no studies exist comparing these two techniques. Objective: The purpose of this study was to compare diagnosis success rates and complications between the suboccipital transcerebellar and transfrontal trajectories. Methods: The study evaluated 142 patients submitted to stereotactic biopsy. The patients presented brainstem tumors in the following areas: pons (n = 31), midbrain (n = 36), medulla (n = 2), pons-medulla (n = 30), pons-midbrain (n = 33), and midbrain-pons-medulla (n = 10). On 123 patients, the transfrontal approach was used, and on 19 the suboccipital transcerebellar approach. Results: Comparing success rates between the two approaches, it was observed that in the group of patients submitted to the transfrontal approach, 95.1% (117 cases) were successful, while in those submitted to the suboccipital transcerebellar approach, 84.2% (16 cases) were successful. Despite a higher success rate among patients in the first group, the difference was not statistically significant. Regarding complications, in patients who were biopsied via the transfrontal trajectory, the morbidity rate was 9.8% (12 cases), while in patients submitted to the suboccipital transcerebellar approach, the morbidity rate was 5.3% (1 case) and the mortality rate 5.3% (1 case). Conclusions: This study verified a higher diagnosis rate in patients submitted to the transfrontal approach than in those submitted to the suboccipital transcerebellar approach (95.1 vs. 84.2%); however, the difference was not statistically significant. Regarding complications, the rate was similar in both groups of patients.
A Retrograde Tracing Study of Compensatory Corticospinal Projections in Rats with Neonatal Hemidecortication
Authors: Yoshikawa Akira , Atobe Yoshitoshi , Takeda Akihito , Kamiya Yoshinori , Takiguchi Masahito , Funakoshi Kengo
Published: January 2012

To examine the compensatory mechanisms in rats that underwent left decortication at postnatal day 7 (P7), we injected the retrograde tracers fluorescein isothiocyanate-cholera toxin B subunit (FITC-CTB) and Fast Blue (FB) into the right and left upper cervical spinal cord, respectively, at postoperative weeks 2, 3, 4, and 5 and counted the number of retrogradely labeled corticospinal neurons in the right cerebral cortex compared with that in normally developed rats. Significantly more ipsilaterally projecting neurons were labeled with FITC-CTB in the decorticated rats compared with normal rats at all time points examined. The number of labeled neurons was similar to that at P7 in normal rats. There were also some FITC-CTB and FB double-labeled neurons in both decorticated and normal rats. The number of double-labeled neurons in the decorticated rats increased each week and was significantly greater than that in normal rats at postoperative weeks 4 and 5. The present results suggest that the elimination of ipsilaterally projecting axons observed in normal rats was prevented in the decorticated rats, so that the cerebral cortex neurons on the unlesioned side projected corticospinal tracts to the ipsilateral spinal cord. Furthermore, the collaterals of the corticospinal tracts originating from the cerebral cortex on the unlesioned side also project to the ipsilateral spinal cord. These compensatory mechanisms might underlie the acquisition of motor function in these animals.
Ultrastructure Study of Transgenic Ren2 Rat Aorta – Part 1: Endothelium and Intima
Authors: Hayden Melvin R. , Habibi Javad , Joginpally Tejaswini , Karuparthi Poorna R. , Sowers James R.
Published: January 2012

Background: The renin-angiotensin-aldosterone system plays an important role in the development and progression of hypertension and accelerated atherosclerosis (atheroscleropathy) associated with the cardiorenal metabolic syndrome and type 2 diabetes mellitus. Additionally, the renin-angiotensin-aldosterone system plays an important role in vascular-endothelial-intimal cellular and extracellular remodeling. Methods: Thoracic aortas of young male transgenic heterozygous (mRen2)27 (Ren2) rats were utilized for this ultrastructural study. This lean model of hypertension, insulin resistance and oxidative stress harbors the mouse renin gene with increased local tissue (aortic) levels of angiotensin II and angiotensin type 1 receptors and elevated plasma aldosterone levels. Results: The ultrastructural observations included marked endothelial cell retraction, separation, terminal nuclear lifting, adjacent duplication, apoptosis and a suggestion of endothelial progenitor cell attachment. The endothelium demonstrated increased caveolae, microparticles, depletion of Weibel-Palade bodies, loss of cell-cell and basal adhesion hemidesmosome-like structures, platelet adhesion and genesis of subendothelial neointima. Conclusion: These observational ultrastructural studies of the transgenic Ren2 vasculature provide an in-depth evaluation of early abnormal remodeling changes within conduit-elastic arteries under conditions of increased local levels of angiotensin II, oxidative stress, insulin resistance and hypertension.
In vitro Activities of the Novel Ceragenin CSA-13, Alone or in Combination with Colistin, Tobramycin, and Ciprofloxacin, against Pseudomonas aeruginosa Strains Isolated from Cystic Fibrosis Patients
Authors: Bozkurt-Guzel Cagla , Savage Paul B. , Gerceker Ayse Alev
Published: January 2012

Background: The rise in the rates of antibiotic resistance among Pseudomonas aeruginosa strains from cystic fibrosis (CF) patients is concerning and underscores the need for the development of novel compounds. Among them CSA-13, a cationic steroid molecule, mimics the activity of naturally occurring antimicrobial peptides. Methods: MICs and MBCs were determined using the microbroth dilution technique. Combinations were assessed using the checkerboard technique. The bactericidal activity of CSA-13 in combination with colistin was measured using the time-kill curve method for two strains. Results: The MIC90 values of CSA-13, colistin, tobramycin, and ciprofloxacin were 2, 1, 1, and 2 mg/l, respectively. The MBCs were equal to or two-fold greater than those of the MICs. With a fractional inhibitory concentration index of ≤0.5 as borderline, synergistic interactions were mostly seen with the CSA-13-colistin combination (54%). No antagonism was observed. The results of the time-kill curve analysis demonstrated rapid bactericidal activity of CSA-13 and synergism with colistin; in one strain early synergy was achieved. Conclusion: CSA-13 appears to be a good candidate in the treatment of P. aeruginosa strains in CF patients. Future studies should be performed to correlate the safety, efficacy, and pharmacokinetic parameters of this molecule.
Recombinant Cell Lines Expressing shRNA Targeting Herpes Simplex Virus 2 VP16 Inhibit Virus Replication
Authors: Zhang Rui , Wang Yan , Song Bo , Han Zhi Qiang , Xu Yu Ming
Published: January 2012

Aims: To establish HSV2 VP16 targeting shRNA-expressing cell lines and investigate the antiviral effect of shRNA targeting HSV2 VP16. Methods: The cell lines Vero-shRNAs and negative-control Vero-shCON were established. Their inhibition effects on VP16 mRNA expression were tested by real-time fluorescent quantitative polymerase chain reaction (PCR) and their antiviral effects were evaluated by yield reduction assay. The influence of passage numbers on the inhibition ability of cell lines was researched. Results: Vero-shRNA24 targeting the upper stream, Vero-shRNA642 targeting the lower stream and Vero-shCON were established. Vero-shRNA24, Vero-shRNA642 and Vero-shRNA24 + 642 could reduce the VP16 mRNA significantly. Vero-shRNA24 was the most efficient. The HSV2 titers in Vero and Vero-shCON were the highest at 72 h after infection, and started decreasing thereafter. The viral titers of the Vero-shRNA groups reached a peak after 84 h and the highest titers were lower than in the Vero group. The inhibiting effect on VP16 mRNA expression and viral replication of Vero-shRNA24 cell lines of passages 10 and 20 were not significantly different from the primary cell line. Although of no statistical significance, the passage 50 cell line showed decreased inhibiting ability. Conclusions: Recombinant cell lines expressing shRNA targeting HSV2 VP16 were established. They can stably inhibit HSV2 VP16 mRNA expression and viral replication within passage 50.
Maternal Serum Placental Growth Factor in Prospective Screening for Aneuploidies at 8–13 Weeks’ Gestation
Authors: Pandya Pranav , Wright David , Syngelaki Argyro , Akolekar Ranjit , Nicolaides Kypros H.
Published: January 2012

Objective: To investigate whether measurement of maternal serum placental growth factor (PLGF) can improve the performance of first-trimester combined screening for trisomy-21 by fetal nuchal translucency (NT) thickness and serum free β-human chorionic gonadotropin (β-hCG) and PAPP-A. Methods: In singleton pregnancies attending for routine care, serum PLGF, free β-hCG and PAPP-A were measured at 8+0–13+6 weeks’ gestation, and fetal NT was measured at 11+0–13+6 weeks. The population included 12,154 normal and 44 trisomy-21 pregnancies. We examined the effect of adding PLGF on the performance of screening by the combined test. Results: In the trisomy-21 pregnancies the median multiple of the normal median PLGF, adjusted for gestational age, maternal weight, racial origin, smoking status and method of conception, was significantly reduced (0.6070, 95% CI 0.5543–0.6648), and this did not change significantly with gestational age. Adding PLGF to combined testing with a risk cut-off of 1 in 100 reduced the false positive rate from 2.7% (95% CI 2.5–3.0) to 2.6% (95% CI 2.4–2.8) and increased the detection rate from 85% (95% CI 75–93) to 88% (95% CI 78–95). Conclusions: Inclusion of serum PLGF improves the performance of the first-trimester combined test in screening for trisomy-21.
Is Awareness of Chronic Kidney Disease Associated with Evidence-Based Guideline-Concordant Outcomes?
Authors: Tuot Delphine S. , Plantinga Laura C. , Hsu Chi-yuan , Powe Neil R.
Published: January 2012

Background: Awareness of chronic kidney disease (CKD) is low. Efforts are underway to increase recognition of CKD among patients, assuming that such an increase will lead to better outcomes through greater adherence to proven therapies. Few studies have tested this assumption. Methods: CKD awareness, defined by a ‘yes’ answer to ‘Have you ever been told by a healthcare provider you have weak or failing kidneys?’, was assessed among 2,404 adults with CKD stages 1–4, who participated in the 2003–2008 National Health and Nutrition Examination Surveys. Odds of blood pressure (BP) control, self-reported use of angiotensin-converting enzyme inhibitors (ACEI) or angiotensin receptor blockers (ARB), and glycemic control, were determined among those aware vs. unaware of their CKD. Results: Optimal BP control, ACEI/ARB use and glycemic control were low in the US adult population with CKD, although there was a recent increase in attainment of guideline-concordant BP control. Odds of BP control and ACEI/ARB use were not different among individuals aware of their CKD compared to those unaware (adjusted odds ratio (AOR) 0.91; 95% CI 0.52–1.58 and AOR 0.75; 0.44–1.30, respectively). CKD awareness among diabetic participants was not associated with glycemic control (AOR 0.41; 95% CI 0.14–1.18). Conclusion: Awareness of CKD is not associated with more optimal BP control, ACEI/ARB use or glycemic control. Future efforts in this area should further explore the measurement of CKD awareness and behaviors associated with CKD awareness.