bluuebunny/crossref_metadata_2025 huggingface dataset:
Single Balloon Enteroscopy for Endoscopic Retrograde Cholangiography in a Patient with Hepaticojejunostomy after Liver Transplant
Authors: Di Pisa Marta , Miraglia Roberto , Volpes Riccardo , Gruttadauria Salvatore , Traina Mario
Published: May 2010

We report a case of a post-transplant patient with hepaticojejunostomy in whom we used a single balloon enteroscopy to access the biliary tree. This procedure seems to be safe and feasible for approaching the biliary anastomosis by means of the overtube and fixation of the small bowel by the balloon.
Effect of Common Buffers and Heterocyclic Ligands on the Binding of Cu(II) at the Multimetal Binding Site in Human Serum Albumin
Authors: Sokołowska Magdalena , Pawlas Krystyna , Bal Wojciech
Published: May 2010

Visible‐range circular dichroism titrations were used to study Cu(II) binding properties of Multimetal Binding Site (MBS) of Human Serum Albumin (HSA). The formation of ternary MBS‐Cu(II)‐Buffer complexes at pH 7.4 was positively verified for sodium phosphate, Tris, and Hepes, the three most common biochemical buffers. The phosphate > Hepes > Tris order of affinities, together with strong spectral changes induced specifically by Tris, indicates the presence of both Buffer‐Cu(II) and Buffer‐HSA interactions. All complexes are strong enough to yield a nearly 100% ternary complex formation in 0.5 mM HSA dissolved in 100 mM solutions of respective buffers. The effects of warfarin and ibuprofen, specific ligands of hydrophobic pockets I and II in HSA on the Cu(II) binding to MBS were also investigated. The effects of ibuprofen were negligible, but warfarin diminished the MBS affinity for Cu(II) by a factor of 20, as a result of indirect conformational effects. These results indicate that metal binding properties of MBS can be modulated directly and indirectly by small molecules.
Coupled Fixed Point Theorems for Nonlinear Contractions Satisfied Mizoguchi-Takahashi's Condition in Quasiordered Metric Spaces
Authors: Du Wei-Shih
Published: May 2010

The main aim of this paper is to study and establish some new coupled fixed point theorems for nonlinear contractive maps that satisfied Mizoguchi-Takahashi's condition in the setting of quasiordered metric spaces or usual metric spaces.
Optical Sensing of Attached Fibrinogen on Carbon Doped Titanium Surfaces
Authors: Silvennoinen Raimo , Vetterl Vladimír , Hasoň Stanislav , Silvennoinen Martti , Myller Kari , Vanĕk Jiři , Cvrček Ladislav
Published: May 2010

The adsorption/desorption of Human Plasma fibrinogen (HPF) molecules on biosurfaces was measured in spectroscopic cuvette by a diffractive optical element- (DOE-) based sensor. To characterize the surfaces, the basic parameters as surface tension was obtained by sensing of a contact angle of water droplet and dielectric constant was measured by ellipsometry in the absence of HPF molecules. It was observed a significant correlation between the adsorption ability of HPF molecules (sensed by DOE on the basis of the changes in optical roughness (Ropt) of studied surface in the absence and presence of HPF molecules), and dielectric constant (measured by ellipsometry) of differently treated titanium surfaces, where the permittivity and dielectric loss have the known linear relation. These findings with carbon-treated biomaterial surfaces can help us to understand mechanisms behind attachment of HPF molecules on biomaterial surfaces to realize and extend variety of implants for hard tissue replacement.
Assessment of Growth Disturbance in Japanese Children with IBD
Authors: Shono Tetsuo , Kato Mayuko , Aoyagi Yo , Haruna Hidenori , Fujii Tohru , Kudo Takahiro , Ohtsuka Yoshikazu , Shimizu Toshiaki
Published: May 2010

In Japan, there is as yet no report on growth retardation in children with IBD. We therefore investigated the cause of growth retardation in Japanese children with IBD. We investigated the height, body weight, serum levels of albumin, IGF-I, CRP, and cytokines, and the amount of corticosteroid administered in children with Crohn's disease (CD, ) and ulcerative colitis (UC, ). Our results suggest that growth retardation is already present before the initial visit in children with CD, and chronic inflammation may be responsible this growth disturbance. Moreover, the amount of PSL used may contribute to growth retardation by decreasing the serum levels of IGF-I in children with IBD.
In vitro Progression of Human Papillomavirus 16 Episome-Associated Cervical Neoplasia Displays Fundamental Similarities to Integrant-Associated Carcinogenesis
Authors: Gray Elizabeth , Pett Mark R. , Ward Dawn , Winder David M. , Stanley Margaret A. , Roberts Ian , Scarpini Cinzia G. , Coleman Nicholas
Published: May 2010

An important event in the development of cervical squamous cell carcinoma (SCC) is deregulated expression of high-risk human papillomavirus (HR-HPV) oncogenes, most commonly related to viral integration into host DNA. Mechanisms of development of the ∼15% of SCCs that contain extrachromosomal (episomal) HR-HPV are poorly understood due to limited longitudinal data. We therefore used the W12 model to study mechanisms of cervical carcinogenesis associated with episomal HPV16. In vitro progression of W12 normally occurs through selection of cells containing integrated HPV16. However, in one long-term culture, keratinocytes developed a selective growth advantage and invasive phenotype while retaining HPV16 episomes at increased copy number in the absence of transcriptionally active integrants. Longitudinal investigations revealed similarities between the episome- and integrant-associated routes of neoplastic progression. Most notable were dynamic changes in viral early gene expression in episome-retaining cells, consistent with continually changing selective pressures. An early increase in viral transcription preceded elevated episome copy number and was followed by a reduction to near baseline after the development of invasiveness. Episomal transcriptional deregulation did not require selection of a specific sequence variant of the HPV16 upstream regulatory region, although increased levels of acetylated histone H4 around the late promoter implicated a role for altered chromatin structure. Interestingly, invasive episome-retaining cells showed high levels of HPV16 E2/E6 proteins (despite decreased transcript levels) and reduced expression of IFN-stimulated genes, adaptations that support viral persistence and cell survival. Our findings suggest a unified working model for events important in cervical neoplastic progression regardless of HR-HPV physical state. Cancer Res; 70(10); 4081–91. ©2010 AACR.
Phospholipase D1 Drives a Positive Feedback Loop to Reinforce the Wnt/β-Catenin/TCF Signaling Axis
Authors: Kang Dong Woo , Lee Soung-Hoon , Yoon Jeong Whan , Park Won-Sang , Choi Kang-Yell , Min Do Sik
Published: May 2010

Activation of the Wnt signaling pathway occurs frequently in human cancers, but an understanding of the targets and regulation of this important pathway remains incomplete. In this study, we report that phospholipase D (PLD), a cell survival mediator that is upregulated in cancer, is an important target of the Wnt signaling pathway that functions in a positive feedback loop to reinforce pathway output. PLD1 expression and activity was enhanced by treatment with Wnt3a and glycogen synthase kinase-3 inhibitors, and the Wnt pathway–regulated transcription factors β-catenin and TCF-4 were required for this effect. Three functional TCF-4–binding sites were identified within the PLD1 promoter. Interestingly, suppressing PLD1 blocked the ability of β-catenin to transcriptionally activate PLD1 and other Wnt target genes by preventing β-catenin/TCF-4 complex formation. Conversely, tactics to elevate intracellular levels of phosphatidic acid, the product of PLD1 enzyme activity, enhanced β-catenin/TCF-4 complex formation as well as β-catenin–dependent TCF transcriptional activity. In cell-based assays, PLD1 was necessary for the anchorage-independent growth driven by Wnt/β-catenin signaling, whereas β-catenin/TCF-4 was necessary for the anchorage-independent growth driven by PLD1 activation. Taken together, our findings define a function for PLD1 in a positive feedback loop of Wnt/β-catenin/TCF-4 signaling that provides new mechanistic insights into cancer, with implications of novel strategies to disrupt Wnt signaling in cancer. Cancer Res; 70(10); 4233–42. ©2010 AACR.
In vivo Dynamics and Distinct Functions of Hypoxia in Primary Tumor Growth and Organotropic Metastasis of Breast Cancer
Authors: Lu Xin , Yan Carol H. , Yuan Min , Wei Yong , Hu Guohong , Kang Yibin
Published: May 2010

Tumor hypoxia is known to activate angiogenesis, anaerobic glycolysis, invasion, and metastasis. However, a comparative analysis of the potentially distinct functions of hypoxia in primary tumor growth and organ-specific metastasis has not been reported. Here, we show distinct hypoxia kinetics in tumors generated by the MDA-MB-231 breast cancer sublines with characteristically different primary tumor growth rates and organotropic metastasis potentials. Hypoxia-induced angiogenesis promotes both primary tumor growth and lung metastasis but is nonessential for bone metastasis. Microarray profiling revealed that hypoxia enhances the expression of a significant number of genes in the lung metastasis signature, but only activates a few bone metastasis genes, among which DUSP1 was functionally validated in this study. Despite the different mechanisms by which hypoxia promotes organ-specific metastasis, inhibition of HIF-1α with a dominant-negative form of HIF-1α or 2-methoxyestradiol reduced metastasis to both lung and bone. Consistent with the extensive functional overlap of hypoxia in promoting primary tumor growth and lung metastasis, a 45-gene hypoxia response signature efficiently stratifies breast cancer patients with low or high risks of lung metastasis, but not for bone metastasis. Our study shows distinct functions of hypoxia in regulating angiogenesis and metastasis in different organ microenvironments and establishes HIF-1α as a promising target for controlling organotropic metastasis of breast cancer. Cancer Res; 70(10); 3905–14. ©2010 AACR.
ΔNp63α Repression of the Notch1 Gene Supports the Proliferative Capacity of Normal Human Keratinocytes and Cervical Cancer Cells
Authors: Yugawa Takashi , Narisawa-Saito Mako , Yoshimatsu Yuki , Haga Kei , Ohno Shin-ichi , Egawa Nagayasu , Fujita Masatoshi , Kiyono Tohru
Published: May 2010

The p53 family member p63 is a master regulator of epithelial development. One of its isoforms, ΔNp63α, is predominantly expressed in the basal cells of stratified epithelia and plays a fundamental role in control of regenerative potential and epithelial integrity. In contrast to p53, p63 is rarely mutated in human cancers, but it is frequently overexpressed in squamous cell carcinomas (SCC). However, its functional relevance to tumorigenesis remains largely unclear. We previously identified the Notch1 gene as a novel transcriptional target of p53. Here, we show that ΔNp63α functions as a transcriptional repressor of the Notch1 gene through the p53-responsive element. Knockdown of p63 caused upregulation of Notch1 expression and marked reduction in proliferation and clonogenicity of both normal human keratinocytes and cervical cancer cell lines overexpressing ΔNp63α. Concomitant silencing of Notch1 significantly rescued this phenotype, indicating the growth defect induced by p63 deficiency to be, at least in part, attributable to Notch1 function. Conversely, overexpression of ΔNp63α decreased basal levels of Notch1, increased proliferative potential of normal human keratinocytes, and inhibited both p53-dependent and p53-independent induction of Notch1 and differentiation markers upon genotoxic stress and serum exposure, respectively. These results suggest that ΔNp63α maintains the self-renewing capacity of normal human keratinocytes and cervical cancer cells partly through transcriptional repression of the Notch1 gene and imply a novel pathogenetical significance of frequently observed overexpression of ΔNp63α together with p53 inactivation in SCCs. Cancer Res; 70(10); 4034–44. ©2010 AACR.
Hypoxia Modulates EWS-FLI1 Transcriptional Signature and Enhances the Malignant Properties of Ewing's Sarcoma Cells In vitro
Authors: Aryee Dave N.T. , Niedan Stephan , Kauer Maximilian , Schwentner Raphaela , Bennani-Baiti Idriss M. , Ban Jozef , Muehlbacher Karin , Kreppel Michael , Walker Robert L. , Meltzer Paul , Poremba Christopher , Kofler Reinhard , Kovar Heinrich
Published: May 2010

Hypoxia is an important condition in the tumor cell microenvironment and approximately 1% to 1.5% of the genome is transcriptionally responsive to hypoxia with hypoxia-inducible factor-1 (HIF-1) as a major mediator of transcriptional activation. Tumor hypoxia is associated with a more aggressive phenotype of many cancers in adults, but data on pediatric tumors are scarce. Because, by immunohistochemistry, HIF-1α expression was readily detectable in 18 of 28 primary Ewing's sarcoma family tumors (ESFT), a group of highly malignant bone-associated tumors in children and young adults, we studied the effect of hypoxia on ESFT cell lines in vitro. Intriguingly, we found that EWS-FLI1 protein expression, which characterizes ESFT, is upregulated by hypoxia in a HIF-1α–dependent manner. Hypoxia modulated the EWS-FLI1 transcriptional signature relative to normoxic conditions. Both synergistic as well as antagonistic transcriptional effects of EWS-FLI1 and of hypoxia were observed. Consistent with alterations in the expression of metastasis-related genes, hypoxia stimulated the invasiveness and soft agar colony formation of ESFT cells in vitro. Our data represent the first transcriptome analysis of hypoxic ESFT cells and identify hypoxia as an important microenvironmental factor modulating EWS-FLI1 expression and target gene activity with far-reaching consequences for the malignant properties of ESFT. Cancer Res; 70(10); 4015–23. ©2010 AACR.
Involvement of the Tubulin Tyrosine Ligase-Like Family Member 4 Polyglutamylase in PELP1 Polyglutamylation and Chromatin Remodeling in Pancreatic Cancer Cells
Authors: Kashiwaya Kotoe , Nakagawa Hidewaki , Hosokawa Masayo , Mochizuki Yasuo , Ueda Koji , Piao Lianhua , Chung Suyoun , Hamamoto Ryuji , Eguchi Hidetoshi , Ohigashi Hiroaki , Ishikawa Osamu , Janke Carsten , Shinomura Yasuhisa , Nakamura Yusuke
Published: May 2010

Polyglutamylation is a new class of posttranslational modification in which glutamate side chains are formed in proteins, although its biological significance is not well known. Through our genome-wide gene expression profile analyses of pancreatic ductal adenocarcinoma (PDAC) cells, we identified the overexpression of tubulin tyrosine ligase-like family member 4 (TTLL4) in PDAC cells. Subsequent reverse transcription-PCR and Northern blot analyses confirmed its upregulation in several PDACs. TTLL4 belongs to the TTLL family which was reported to have polyglutamylase activity. Knockdown of TTLL4 by short hairpin RNA in PDAC cells attenuated the growth of PDAC cells and exogenous introduction of TTLL4 enhanced cell growth. We also found that TTLL4 expression was correlated with polyglutamylation levels of a glutamate stretch region of the proline, glutamate, and leucine–rich protein 1 (PELP1) that was shown to interact with various proteins such as histone H3, and was involved in several signaling pathways through its function as a scaffold protein. PELP1 polyglutamylation could influence its interaction with histone H3 and affect histone H3 acetylation. We also identified the interaction of PELP1 with LAS1L and SENP3, components of the MLL1-WDR5 supercomplex involving chromatin remodeling. Our findings imply that TTLL4 could play important roles in pancreatic carcinogenesis through its polyglutamylase activity and subsequent coordination of chromatin remodeling, and might be a good molecular candidate for the development of new therapeutic strategies for pancreatic cancer. Cancer Res; 70(10); 4024–33. ©2010 AACR.
The α-Receptor for Platelet-Derived Growth Factor Confers Bone-Metastatic Potential to Prostate Cancer Cells by Ligand- and Dimerization-Independent Mechanisms
Authors: Russell Mike R. , Liu Qingxin , Lei Hetian , Kazlauskas Andrius , Fatatis Alessandro
Published: May 2010

Prostate adenocarcinoma is the second leading cause of cancer death among men, due primarily to the fact that the majority of prostate cancers will eventually spread to the skeleton. Metastatic dissemination requires a complex series of coordinated events that result in cells that escape from the primary tumor into the circulation and eventually colonize a distant organ. The ability of these cells to evolve into macroscopic metastases depends strongly on their compatibility with, and ability to utilize, this new microenvironment. We previously showed that bone-metastatic prostate cancer cells exposed to human bone marrow respond by activation of cell survival pathways, such as phosphoinositide 3-kinase/Akt, and that these events are mediated by the α-receptor for platelet-derived growth factor (PDGFRα). Our studies and others have shown that PDGFRα may be activated by mechanisms independent of PDGF ligand binding. Here, we provide conclusive evidence that soluble components of human bone marrow can activate PDGFRα through a mechanism that does not require the canonical binding of PDGF ligand(s) to the receptor. In particular, we found that dimerization of PDGFRα monomers is not induced by human bone marrow, but this does not prevent receptor phosphorylation and downstream signaling from occurring. To establish the relevance of this phenomenon in vivo, we used a PDGFRα mutant lacking the extracellular ligand-binding domain. Our studies show that this truncated PDGFRα is able to restore bone-metastatic potential of prostate cancer cells as effectively as the full-length form of the receptor. Cancer Res; 70(10); 4195–203. ©2010 AACR.
Association of SPINK1 Expression and TMPRSS2:ERG Fusion with Prognosis in Endocrine-Treated Prostate Cancer
Authors: Leinonen Katri A. , Tolonen Teemu T. , Bracken Hazel , Stenman Ulf-Håkan , Tammela Teuvo L.J. , Saramäki Outi R. , Visakorpi Tapio
Published: May 2010

Purpose: The aim of the study was to examine whether TMPRSS2:ERG fusion or SPINK1 protein expression is associated with hormone responsiveness of prostate cancer and can thus be used as a biomarker.

Experimental Design: Diagnostic needle biopsies from prostate cancer patients primarily treated by endocrine therapy were evaluated for TMPRSS2:ERG fusion with fluorescence in situ hybridization and SPINK1 protein expression with immunohistochemistry.

Results: The frequency of TMPRSS2:ERG fusion in 178 biopsies of hormonally treated patients was 34%. Of the fusion-positive cases, 71% showed deletion between the two genes, and 23% showed gain of the fusion. The fusion was associated with high Ki-67 staining (P = 0.001), age at diagnosis (P = 0.024), and tumor area (P = 0.006), but not with Gleason score, T stage, M stage, prostate-specific antigen (PSA), or progression-free survival. Strong positive SPINK1 expression was found in 11% (21 of 186) of the biopsies. SPINK1-positive cases had significantly shorter progression-free survival compared with SPINK1-negative cases (P = 0.001). The expression was not associated with any other clinicopathologic variables studied. In a multivariate analysis, SPINK1 expression showed independent prognostic value, with a relative risk of 2.3 (95% confidence interval, 1.1-4.6). SPINK1 expression and the fusion were not associated with each other.

Conclusions: There was no association between TMPRSS2:ERG fusion and prognosis, suggesting that TMPRSS2:ERG rearrangement does not implicate hormone dependence of the cancer. SPINK1 expression, found in ∼10% of prostate cancers, was associated with aggressive form of the disease and could serve as a biomarker in endocrine-treated prostate cancer. Clin Cancer Res; 16(10); 2845–51. ©2010 AACR.
Inhibition of Histone Deacetylase Attenuates Hypoxia-Induced Migration and Invasion of Cancer Cells via the Restoration of RECK Expression
Authors: Jeon Hye Won , Lee You Mie
Published: May 2010

Hypoxia is a strong signal for cell migration and invasion in cancer. The reversion-inducing cysteine-rich protein with Kazal motif (RECK), a tumor suppressor, inhibits cancer cell migration and invasion and is frequently silenced in aggressive tumor cells by histone deacetylases (HDAC). However, the effect of RECK silencing in several cancer cells in a hypoxic microenvironment has not been fully delineated. In this report, we investigated whether hypoxia suppressed RECK expression and used HDAC inhibitor (HDACI) inhibition to restore RECK expression to inhibit cancer cell migration and invasion. HDACIs, including trichostatin A (TSA), completely rescued RECK expression, which was suppressed by hypoxia, in the H-Ras–transformed human breast MCF10A and the HT1080 cell lines (human fibrosarcoma). TSA suppressed the activity of matrix metalloproteinase-2 (MMP-2) and MMP-9, induced by hypoxia, and significantly inhibited hypoxia-stimulated migration and invasion of both cancer cells. RECK overexpression significantly inhibited the migration and invasion of cancer cells induced by hypoxia. The hypoxic effect on the migration and invasion of cells was equivalent to the effect seen using the small interfering RNA (siRNA) of RECK under normoxia, suggesting an inhibitory role for RECK in hypoxic conditions. We also showed that siRNA silencing of HDAC1 suppressed hypoxia-induced RECK downregulation and inhibited the migration and invasion of cancer cells. In conclusion, the inhibition of HDAC successfully restored the expression of RECK under hypoxic conditions. This resulted in the inhibition of cancer cell migration and invasion through the repression of MMP-2 and MMP-9 activity. Mol Cancer Ther; 9(5); 1361–70. ©2010 AACR.
Expression of Oncogenic Kinase Bcr-Abl Impairs Mitotic Checkpoint and Promotes Aberrant Divisions and Resistance to Microtubule-Targeting Agents
Authors: Wolanin Kamila , Magalska Adriana , Kusio-Kobialka Monika , Podszywalow-Bartnicka Paulina , Vejda Susanne , McKenna Sharon L. , Mosieniak Grazyna , Sikora Ewa , Piwocka Katarzyna
Published: May 2010

Recent findings showed that BRCA1, in addition to its role in DNA damage response, acts as an upstream regulator of genes involved in the mitotic checkpoint regulation, thus protecting against promotion of aberrant divisions and aneuploidy. Moreover, there is also an indication that the BRCA1 protein is downregulated in chronic myeloid leukemia (CML) patients. We have investigated a possible functional relationship between BRCA1 and mitotic checkpoint competence in cells with the same genetic background expressing different levels of Bcr-Abl, an oncogene responsible for CML. Herein, we show that Bcr-Abl strongly downregulates the BRCA1 protein level, which is partially reversed on treatment with imatinib, an inhibitor of Bcr-Abl tyrosine kinase. Bcr-Abl leads to decreased expression of genes involved in the mitotic checkpoint activation—Mad2, Bub1, Bub3, and BubR1, resulting in mitosis perturbances, weakened mitotic checkpoint function, and mitotic slippage after nocodazole treatment. Furthermore, high Bcr-Abl–expressing cells showed also postmitotic checkpoint dysfunctions and inability to effectively arrest in the 4NG1 phase of the cell cycle, which was associated with limited p21 induction. These observations had significant biological consequences, as we found a high level of improper divisions, chromosomal missegregation, and generation of polyploid cells on mitotic checkpoint prolonged activation. Additionally, Bcr-Abl–expressing cells showed resistance to death activated by spindle defects, reversed by imatinib. Our study presents new facts and supports the hypothesis concerning the mutator nature of Bcr-Abl itself. The functional interaction between Bcr-Abl and mitosis dysfunctions, due to compromised mitotic checkpoints, may have important implications for the generation of aneuploidy and CML progression. Mol Cancer Ther; 9(5); 1328–38. ©2010 AACR.
S110, a 5-Aza-2′-Deoxycytidine–Containing Dinucleotide, Is an Effective DNA Methylation Inhibitor In vivo and Can Reduce Tumor Growth
Authors: Chuang Jody C. , Warner Steven L. , Vollmer David , Vankayalapati Hariprasad , Redkar Sanjeev , Bearss David J. , Qiu Xiangning , Yoo Christine B. , Jones Peter A.
Published: May 2010

Methylation of CpG islands in promoter regions is often associated with gene silencing and aberrant DNA methylation occurs in most cancers, leading to the silencing of some tumor suppressor genes. Reversal of this abnormal hypermethylation by DNA methylation inhibitors is effective in reactivating methylation-silenced tumor suppressor genes both in vitro and in vivo. Several DNA methylation inhibitors have been well studied; the most potent among them is 5-aza-2′-deoxycytidine (5-Aza-CdR), which can induce myelosuppression in patients. S110 is a dinucleotide consisting of 5-Aza-CdR followed by a deoxyguanosine, which we previously showed to be effective in vitro as a DNA methylation inhibitor while being less prone to deamination by cytidine deaminase, making it a promising alternative to 5-Aza-CdR. Here, we show that S110 is better tolerated than 5-Aza-CdR in mice and is as effective in vivo in inducing p16 expression, reducing DNA methylation at the p16 promoter region, and retarding tumor growth in human xenograft. We also show that S110 is effective by both i.p. and s.c. deliveries. S110 therefore is a promising new agent that acts similarly to 5-Aza-CdR and has better stability and less toxicity. Mol Cancer Ther; 9(5); 1443–50. ©2010 AACR.
A Novel Small-Molecule Inhibitor of Protein Kinase D Blocks Pancreatic Cancer GrowthIn vitroandIn vivo
Authors: Harikumar Kuzhuvelil B. , Kunnumakkara Ajaikumar B. , Ochi Nobuo , Tong Zhimin , Deorukhkar Amit , Sung Bokyung , Kelland Lloyd , Jamieson Stephen , Sutherland Rachel , Raynham Tony , Charles Mark , Bagherzadeh Azadeh , Foxton Caroline , Boakes Alexandra , Farooq Muddasar , Maru Dipen , Diagaradjane Parmeswaran , Matsuo Yoichi , Sinnett-Smith James , Gelovani Juri , Krishnan Sunil , Aggarwal Bharat B. , Rozengurt Enrique , Ireson Christopher R. , Guha Sushovan
Published: May 2010

Protein kinase D (PKD) family members are increasingly implicated in multiple normal and abnormal biological functions, including signaling pathways that promote mitogenesis in pancreatic cancer. However, nothing is known about the effects of targeting PKD in pancreatic cancer. Our PKD inhibitor discovery program identified CRT0066101 as a specific inhibitor of all PKD isoforms. The aim of our study was to determine the effects of CRT0066101 in pancreatic cancer. Initially, we showed that autophosphorylated PKD1 and PKD2 (activated PKD1/2) are significantly upregulated in pancreatic cancer and that PKD1/2 are expressed in multiple pancreatic cancer cell lines. Using Panc-1 as a model system, we showed that CRT0066101 reduced bromodeoxyuridine incorporation; increased apoptosis; blocked neurotensin-induced PKD1/2 activation; reduced neurotensin-induced, PKD-mediated Hsp27 phosphorylation; attenuated PKD1-mediated NF-κB activation; and abrogated the expression of NF-κB-dependent proliferative and prosurvival proteins. We showed that CRT0066101 given orally (80 mg/kg/d) for 24 days significantly abrogated pancreatic cancer growth in Panc-1 subcutaneous xenograft model. Activated PKD1/2 expression in the treated tumor explants was significantly inhibited with peak tumor concentration (12 μmol/L) of CRT0066101 achieved within 2 hours after oral administration. Further, we showed that CRT0066101 given orally (80 mg/kg/d) for 21 days in Panc-1 orthotopic model potently blocked tumor growth in vivo. CRT0066101 significantly reduced Ki-67–positive proliferation index (P < 0.01), increased terminal deoxynucleotidyl transferase–mediated dUTP nick end labeling–positive apoptotic cells (P < 0.05), and abrogated the expression of NF-κB–dependent proteins including cyclin D1, survivin, and cIAP-1. Our results show for the first time that a PKD-specific small-molecule inhibitor CRT0066101 blocks pancreatic cancer growth in vivo and show that PKD is a novel therapeutic target in pancreatic cancer. Mol Cancer Ther; 9(5); 1136–46. ©2010 AACR.
Metformin in Cancer Therapy: A New Perspective for an Old Antidiabetic Drug?
Authors: Ben Sahra Issam , Le Marchand-Brustel Yannick , Tanti Jean-François , Bost Frédéric
Published: May 2010

Metformin is the most widely used antidiabetic drug in the world, and there is increasing evidence of a potential efficacy of this agent as an anticancer drug. First, epidemiological studies show a decrease in cancer incidence in metformin-treated patients. Second, metformin decreases insulin resistance and indirectly reduces insulin level, a beneficial effect because insulin promotes cancer cell growth. Third, several reports outline a direct inhibitory effect of metformin on cancer cell growth and an antitumoral action. Finally, metformin activates the AMP activated protein kinase (AMPK) pathway, a major sensor of the energetic status of the cell, which has been proposed as a promising therapeutic target in cancer. Mol Cancer Ther; 9(5); 1092–99. ©2010 AACR.
Anticancer Efficacy of a Difluorodiarylidenyl Piperidone (HO-3867) in Human Ovarian Cancer Cells and Tumor Xenografts
Authors: Selvendiran Karuppaiyah , Tong Liyue , Bratasz Anna , Kuppusamy M. Lakshmi , Ahmed Shabnam , Ravi Yazhini , Trigg Nancy J. , Rivera Brian K. , Kálai Tamás , Hideg Kálmán , Kuppusamy Periannan
Published: May 2010

The purpose of this study was to evaluate the anticancer potency and mechanism of a novel difluorodiarylidenyl piperidone (H-4073) and its N-hydroxypyrroline modification (HO-3867) in human ovarian cancer. Studies were done using established human ovarian cancer cell lines (A2870, A2780cDDP, OV-4, SKOV3, PA-1, and OVCAR3) as well as in a murine xenograft tumor (A2780) model. Both compounds were comparably and significantly cytotoxic to A2780 cells. However, HO-3867 showed a preferential toxicity toward ovarian cancer cells while sparing healthy cells. HO-3867 induced G2-M cell cycle arrest in A2780 cells by modulating cell cycle regulatory molecules p53, p21, p27, cyclin-dependent kinase 2, and cyclin, and promoted apoptosis by caspase-8 and caspase-3 activation. It also caused an increase in the expression of functional Fas/CD95 and decreases in signal transducers and activators of transcription 3 (STAT3; Tyr705) and JAK1 phosphorylation. There was a significant reduction in STAT3 downstream target protein levels including Bcl-xL, Bcl-2, survivin, and vascular endothelial growth factor, suggesting that HO-3867 exposure disrupted the JAK/STAT3 signaling pathway. In addition, HO-3867 significantly inhibited the growth of the ovarian xenografted tumors in a dosage-dependent manner without any apparent toxicity. Western blot analysis of the xenograft tumor tissues showed that HO-3867 inhibited pSTAT3 (Tyr705 and Ser727) and JAK1 and increased apoptotic markers cleaved caspase-3 and poly ADP ribose polymerase. HO-3867 exhibited significant cytotoxicity toward ovarian cancer cells by inhibition of the JAK/STAT3 signaling pathway. The study suggested that HO-3867 may be useful as a safe and effective anticancer agent for ovarian cancer therapy. Mol Cancer Ther; 9(5); 1169–79. ©2010 AACR.
Thrombin Regulates the Metastatic Potential of Human Rhabdomyosarcoma Cells: Distinct Role of PAR1 and PAR3 Signaling
Authors: Wysoczynski Marcin , Liu Rui , Kucia Magda , Drukala Justyna , Ratajczak Mariusz Z.
Published: May 2010

We observed that human rhabdomyosarcoma (RMS) cells highly express a tissue factor that promotes thrombin formation, which indirectly and directly affects RMS progression. First, we found that thrombin activates platelets to generate microvesicles (PMV), which transfer to RMS cells' α2β3 integrin and increase their adhesiveness to endothelial cells. Accordingly, RMS cells covered with PMVs showed higher metastatic potential after i.v. injection into immunodeficient mice. Furthermore, PMVs activate mitogen-activated protein kinase (MAPK)p42/44 and AKT to chemoattract RMS cells. We also found that RMS cells express functional protease-activated receptor-1 (PAR1) and PAR3 and respond to thrombin stimulation by MAPKp42/44 and MAPKp38 phosphorylation. To our surprise, thrombin did not affect RMS proliferation or survival; it inhibited the chemotactic and adhesive properties of RMS cells. However, when PAR1-specific agonist thrombin receptor–activating peptide 6 was used, which does not activate PAR3, selective PAR1 stimulation enhanced RMS proliferation. To learn more on the role of PAR1 and PAR3 antagonism in RMS proliferation and metastasis, we knocked down both receptors by using a short hairpin RNA strategy. We found that although thrombin does not affect growth of PAR1−/− cells, it stimulated the proliferation of PAR3−/− cells. More importantly, PAR3−/− cells, in contrast to PAR1−/− ones, formed larger tumors in immunodeficient mice. We conclude that thrombin is a novel underappreciated modulator of RMS metastasis and that we have identified a novel role for PAR3 in thrombin signaling. Mol Cancer Res; 8(5); 677–90. ©2010 AACR.
Bortezomib Sensitizes Human Renal Cell Carcinomas to TRAIL Apoptosis through Increased Activation of Caspase-8 in the Death-Inducing Signaling Complex
Authors: Brooks Alan D. , Jacobsen Kristen M. , Li Wenqing , Shanker Anil , Sayers Thomas J.
Published: May 2010

Bortezomib (VELCADE) could sensitize certain human renal cell carcinoma (RCC) lines to the apoptotic effects of tumor necrosis factor–related apoptosis-inducing ligand (TRAIL). Analysis of seven human RCC showed a clear increase in the sensitivity of four of the RCC to TRAIL cytotoxicity following bortezomib (5-20 nmol/L) treatment, whereas the remaining three remained resistant. Tumor cell death following sensitization had all the features of apoptosis. The enhanced antitumor activity of the bortezomib and TRAIL combination was confirmed in long-term (6 days) cancer cell outgrowth assays. The extent of proteasome inhibition by bortezomib in the various RCC was equivalent. Following bortezomib treatment, neither changes in the intracellular protein levels of various Bcl-2 and IAP family members, nor minor changes in expression of TRAIL receptors (DR4, DR5), correlated well with the sensitization or resistance of RCC to TRAIL-mediated apoptosis. However, enhanced procaspase-8 activation following bortezomib pretreatment and subsequent TRAIL exposure was only observed in the sensitized RCC in both cell extracts and death-inducing signaling complex immunoprecipitates. These data suggest that the molecular basis for bortezomib sensitization of RCC to TRAIL primarily involves early amplification of caspase-8 activity. In the absence of this increased caspase-8 activation, other bortezomib-induced changes are not sufficient to sensitize RCC to TRAIL-mediated apoptosis. Mol Cancer Res; 8(5); 729–38. ©2010 AACR.
Clinical Features of Rapidly Progressive Alzheimer’s Disease
Authors: Schmidt Christian , Redyk Katharina , Meissner Bettina , Krack Lennart , von Ahsen Nico , Roeber Sigrun , Kretzschmar Hans , Zerr Inga
Published: May 2010

Objective: To characterize clinical features, CSF biomarkers and genetic polymorphisms of patients suffering from a rapidly progressing subtype of Alzheimer’s dementia (rpAD). Methods: Retrospective analyses of 32 neuropathologically confirmed cases differentially diagnosed as AD out of a group with rapidly progressive dementia. CSF biomarkers (14-3-3, tau, β-amyloid 1–42) and genetic markers (PRNP codon 129, apolipoprotein E, ApoE, polymorphism) were determined. Results: Median survival was 26 months, age at onset 73 years. Biomarkers: mean β-amyloid 1–42: 266 pg/ml, median tau: 491 pg/ml, 14-3-3 positive: 31%. Genetic polymorphisms showed a predominance of methionine homozygosity at PRNP codon 129 and a low frequency of ApoE4 (38%, no homozygous patients). Thirty-five symptoms were studied. Frequent symptoms were myoclonus (75%), disturbed gait (66%) and rigidity (50%). Discussion: rpAD is associated with a diversity of neurological signs even able to mimic Creutzfeldt-Jakob disease. Biomarkers and genetic profile differ from those seen in classical AD. The findings on biomarkers, symptomatology and genetics may aid the differential diagnostic process.
Caspase-Cleaved Transactivation Response DNA-Binding Protein 43 in Parkinson’s Disease and Dementia with Lewy Bodies
Authors: Kokoulina Polina , Rohn Troy T.
Published: May 2010

Background: Transactivation response DNA-binding protein 43 (TDP-43) proteinopathies are classified based upon the extent of modified TDP-43 and include a growing number of neurodegenerative diseases such as amyotrophic lateral sclerosis, frontotemporal lobar degeneration with ubiquitin-immunoreactive, tau-negative inclusions and frontotemporal lobar degeneration with motor neuron disease. Objective: The purpose of the study was to examine whether proteolytic modifications of TDP-43 are a relevant finding in Parkinson’s disease (PD) and dementia with Lewy bodies (DLB). Methods: A novel site-directed caspase cleavage antibody, termed TDP caspase cleavage product antibody (TDPccp), was utilized based upon a known caspase 3 cleavage consensus site within TDP-43 at position 219. Results: Application of this antibody to postmortem brain sections from PD and DLB patients revealed the presence of caspase-cleaved TDP-43 in Lewy bodies and Hirano bodies in all cases examined. Colocalization of TDPccp with an antibody to α-synuclein (α-Syn), which served as a general marker for Lewy bodies, was evident within the substantia nigra in both α-synucleinopathies. Interestingly, the TDPccp antibody detected a greater number of Lewy bodies in PD and DLB compared to the α-Syn antibody. In addition, a semiquantitative analysis in both diseases confirmed this finding by indicating that the percentage of caspase-cleaved TDP-43 single-labeled Lewy bodies was approximately twice that of α-Syn labeling (in DLB 13.4 vs. 5.5%, while in PD 34.6 vs. 17.6%). Conclusion: Collectively, these data have identified caspase-cleaved TDP-43 as a primary component of Lewy and Hirano bodies in PD and DLB, and suggest that the TDPccp antibody is an effective marker for the detection of Lewy bodies in these neurodegenerative diseases.
Cerebrospinal Fluid Concentrations of Functionally Important Amino Acids and Metabolic Compounds in Patients with Mild Cognitive Impairment and Alzheimer’s Disease
Authors: Kaiser Elmar , Schoenknecht Peter , Kassner Stefan , Hildebrandt Wulf , Kinscherf Ralf , Schroeder Johannes
Published: May 2010

Cerebrospinal fluid (CSF) biomarkers play an important role in the differential diagnosis of neurodegenerative diseases such as Alzheimer’s disease (AD) and its postulated precursor stage mild cognitive impairment (MCI). While CSF tau protein, phospho-tau protein and β-amyloid have become part of the diagnostic process in clinical routine, the importance of several other biomarkers remains quite unclear. Among these, amino acids and metabolic compounds have been studied in clinical conditions mostly other than AD and, to our knowledge, never in MCI. In patients with AD (n = 14) and MCI (n = 13) we now determined CSF levels of 36 different amino acids and metabolic compounds by high-performance liquid chromatography. We found that 8 out of 36 amino acids (urea, threonine, glutamate, citrulline, α-aminobutyric acid, ornithine, ammonia and arginine) were significantly elevated in the CSF of patients with AD compared to those with MCI. As most of these amino acids and metabolic compounds are functionally important for brain-specific metabolic processes, neurotransmitter pathways or compensatory mechanisms, our findings might reflect these changes occurring within the brain of patients with MCI and those who developed manifest AD.
Diet and Paraoxonase 1 Enzymatic Activity in Diabetic Foot Patients from Romania and Belgium: Favorable Association of High Flavonoid Dietary Intake with Arylesterase Activity
Authors: Lixandru Daniela , Mohora Maria , Coman Anca , Stoian Irina , van Gils Carolien , Aerts Petra , Manuel-y-Keenoy Begoña
Published: May 2010

Background/Aims: The antiatherosclerotic enzyme paraoxonase (PON1) is affected by disease and lifestyle. We investigated the impact of diet in diabetic foot patients from 2 European countries. Methods: Dietary intake and serum PON1 activity, using as substrate paraoxon (paraoxonase) or phenylacetate (arylesterase), were assessed in patients from Bucharest (n = 40) and Antwerp (n = 30) and in 34 healthy controls. Results: The diabetic patients had lower paraoxonase and arylesterase activities than the controls. Arylesterase was lowest in the Bucharest patients, 116 ± 42 U/ml, versus 141 ± 43 and 184 ± 49 U/ml in the Antwerp patients and controls, respectively (p < 0.0005). The Bucharest patients had worse glycemic control, higher blood pressure, lower HDL cholesterol and a diet richer in cholesterol and poorer in monounsaturated fats and fish. In contrast, their median intake of vitamins E and C, folic acid and flavonoids was higher, 82 mg (range: 4–259 mg), versus 28 mg (range: 5–169 mg) aglycone units in Antwerp (p = 0.005). Flavonoid intake predicted arylesterase independently of HDL cholesterol, region and sex (β = 0.27; p = 0.03), and patients with high intake achieved normal levels of arylesterase (30.1 ± 10.0 U/µmol in the highest versus 21.0 ± 8.2 U/µmol total cholesterol in the lowest tertile; p = 0.02). Conclusion: A flavonoid-rich diet is positively associated with PON1 arylesterase activity in diabetic foot patients.
Radial Forearm Free Flap for Reconstruction of a Large Defect after Radical Ablation of Carcinoma of the Tongue and Floor of the Mouth: Some New Modifications
Authors: Song Xiao-meng , Ye Jin-hai , Yuan Ye , Zhang Shuang-yue , Jiang Hong-bing , Wu Yu-nong
Published: May 2010

Background: A modified radial forearm free flap was designed to rehabilitate function and to reduce the complications at both donor and recipient sites. Methods: Between 2003 and 2007, 15 patients with infiltrating squamous cell carcinoma (T3–T4) of the tongue and/or floor of the mouth underwent hemiglossectomy and resection of the floor of the mouth with microvascular reconstruction using a modified radial forearm flap. The mean size of the forearm flap was 7.5 × 14 cm, and the de-epithelialized area was 7 × 6 cm, requiring no skin graft from the abdomen. Speech intelligibility tests were administered to test postoperative speech and the functional oral intake scale was applied to assess the postoperative swallowing function, and patients reconstructed with pectoralis major myocutaneous flap were used for comparison. Results: All the flaps were successfully transferred. No obvious complications were found in either the oral-maxillofacial or forearm region. The speech intelligibility was better in the modified flap group (p < 0.01). An acceptable swallowing function was also achieved, although the difference was not significant (p > 0.05). Conclusions: The modified flap used for reconstructing large defects of the tongue and floor of the mouth might be a valid substitute for pectoralis major myocutaneous flap to improve the outcome in individuals with significant oral carcinoma.
Novel Method for Controlling Cerebrospinal Fluid Flow and Intracranial Pressure by Use of a Tandem Shunt Valve System
Authors: Aihara Yasuo , Kawamata Takakazu , Mitsuyama Tetsuryu , Hori Tomokatsu , Okada Yoshikazu
Published: May 2010

Objective: Programmable shunt valve systems have improved the management of patients with hydrocephalus. However, associated problems such as over-drainage and slit-like ventricles, even in patients with anti-siphon devices installed, still remain unresolved. Methods: We conceived a novel tandem shunt valve system to overcome the problems with existing shunt valve systems. Experiments were performed in vitro and a manometer was used to measure the results. Results: Manometric measurements in vitro with the tandem shunt valve system demonstrated that it effectively controlled the final pressure under varying flow rates. This novel system was applied to 2 intractable cases (over-drainage and slit-like ventricles) with good results. Interpretation: The tandem shunt valve system has therapeutic potential not only for over-drainage and slit-like ventricles, but also for uncontrollable hydrocephalus, by precisely controlling the cerebral spinal fluid flow rate.
Healthcare Savings Associated with Reduced Infection Rates Using Antimicrobial Suture Wound Closure for Cerebrospinal Fluid Shunt Procedures
Authors: Stone Jonathan , Gruber Thomas J. , Rozzelle Curtis J.
Published: May 2010

Background/Aims/Methods: This is a follow-up study from a recent randomized controlled trial conducted at the Women and Children’s Hospital of Buffalo that investigated the use of antimicrobial sutures (AMS) for wound closure during cerebrospinal fluid shunting procedures. Our purpose was to determine the average cost of shunt infections at our institution and estimate the healthcare savings associated with reduced infection rates secondary to AMS use. Result: We found that the treatment and management of a shunt infection resulted in a 5.3-fold increase in hospital charges. Conclusion: The potential healthcare savings associated with reduced infection rates are significant. These calculations demonstrate the profound economic effect a reduction in shunt infection rates may have on a healthcare system.
Cystic Spinal Dysraphism of the Cervical Region: Experience with Eight Cases Including Double Cervical and Lumbosacral Meningoceles
Authors: Ali Mostafa Z.
Published: May 2010

Objective: Cervical meningoceles are rare. Clinical, radiological and surgical data of 8 cases were presented, including a unique case of double cervical and lumbosacral meningoceles. Methods: This retrospective study included all children operated on for cervical meningocele from January 2004 to June 2009 at the Aburish Pediatric Hospital, Cairo University. Results: Eight children (6 boys and 2 girls) were operated on. Their ages ranged between 2 months and 8 years. The clinical picture – apart form posterior cervical swelling – was almost normal. One case had paraplegia due to associated lumbosacral myelomeningocele (double meningocele). Four patients had associated hydrocephalus; 2 had Chiari malformation and 2 hydromyelia. Conclusion: The outcome after surgery for these lesions is excellent as the majority of the children have no or minimal neurological deficit and surgery does help in improving cosmesis and preventing the development of neurological deterioration.