bluuebunny/crossref_metadata_2025 huggingface dataset:
Wnt Inhibitor Dickkopf-1 as a Target for Passive Cancer Immunotherapy
Authors: Sato Nagato , Yamabuki Takumi , Takano Atsushi , Koinuma Junkichi , Aragaki Masato , Masuda Ken , Ishikawa Nobuhisa , Kohno Nobuoki , Ito Hiroyuki , Miyamoto Masaki , Nakayama Haruhiko , Miyagi Yohei , Tsuchiya Eiju , Kondo Satoshi , Nakamura Yusuke , Daigo Yataro
Published: June 2010

Dickkopf-1 (DKK1) is an inhibitor of Wnt/β-catenin signaling that is overexpressed in most lung and esophageal cancers. Here, we show its utility as a serum biomarker for a wide range of human cancers, and we offer evidence favoring the potential application of anti-DKK1 antibodies for cancer treatment. Using an original ELISA system, high levels of DKK1 protein were found in serologic samples from 906 patients with cancers of the pancreas, stomach, liver, bile duct, breast, and cervix, which also showed elevated expression levels of DKK1. Additionally, anti-DKK1 antibody inhibited the invasive activity and the growth of cancer cells in vitro and suppressed the growth of engrafted tumors in vivo. Tumor tissues treated with anti-DKK1 displayed significant fibrotic changes and a decrease in viable cancer cells without apparent toxicity in mice. Our findings suggest DKK1 as a serum biomarker for screening against a variety of cancers, and anti-DKK1 antibodies as potential theranostic tools for diagnosis and treatment of cancer. Cancer Res; 70(13); 5326–36. ©2010 AACR.
Pancreatic Cancers Epigenetically Silence SIP1 and Hypomethylate and Overexpress miR-200a/200b in Association with Elevated Circulating miR-200a and miR-200b Levels
Authors: Li Ang , Omura Noriyuki , Hong Seung-Mo , Vincent Audrey , Walter Kimberly , Griffith Margaret , Borges Michael , Goggins Michael
Published: June 2010

Aberrant DNA methylation and microRNA expression play important roles in the pathogenesis of pancreatic cancer. While interrogating differentially methylated CpG islands in pancreatic cancer, we identified two members of miR-200 family, miR-200a and miR-200b, that were hypomethylated and overexpressed in pancreatic cancer. We also identified prevalent hypermethylation and silencing of one of their downstream targets, SIP1 (ZFHX1B, ZEB2), whose protein product suppresses E-cadherin expression and contributes to epithelial mesenchymal transition. In a panel of 23 pancreatic cell lines, we observed a reciprocal correlation between miR-200, SIP1, and E-cadherin expression, with pancreatic cancer–associated fibroblasts showing the opposite expression pattern to most pancreatic cancers. In Panc-1 cells, which express SIP1, have low E-cadherin expression, and do not express miR-200a or miR-200b, treatment with miR-200a and miR-200b downregulated SIP1 mRNA and increased E-cadherin expression. However, most pancreatic cancers express miR-200a and miR-200b, but this expression does not affect SIP1 expression, as the SIP1 promoter is silenced by hypermethylation and in these cancers E-cadherin is generally expressed. Both miR-200a and miR-200b were significantly elevated in the sera of pancreatic cancer and chronic pancreatitis patients compared with healthy controls (P < 0.0001), yielding receiver operating characteristic curve areas of 0.861 and 0.85, respectively. In conclusion, most pancreatic cancers display hypomethylation and overexpression of miR-200a and miR-200b, silencing of SIP1 by promoter methylation, and retention of E-cadherin expression. The elevated serum levels of miR-200a and miR-200b in most patients with pancreatic cancer could have diagnostic utility. Cancer Res; 70(13); 5226–37. ©2010 AACR.
Shedding of RANKL by Tumor-Associated MT1-MMP Activates Src-Dependent Prostate Cancer Cell Migration
Authors: Sabbota Aaron L. , Kim Hyeong-Reh Choi , Zhe Xiaoning , Fridman Rafael , Bonfil R. Daniel , Cher Michael L.
Published: June 2010

Membrane type 1 matrix metalloproteinase (MT1-MMP) plays an essential role in protease-mediated extracellular matrix (ECM) degradation, but it also functions as a sheddase releasing non-ECM substrates such as receptor activator of NF-κB ligand (RANKL), an osteoclastogenic factor typically confined to the surface of osteoblasts. We previously found high expression of MT1-MMP in skeletal metastasis of prostate cancer patients, in a pattern similar to RANKL expression. We also showed that overexpression of MT1-MMP in prostate cancer cells increases tumor growth and osteolysis in an intratibial mouse model of bone metastasis, and that soluble factor(s) shed by tumor-derived MT1-MMP enhance osteoclast differentiation in a RANKL-dependent manner. Recent evidence indicates that the cognate receptor for RANKL, RANK, is expressed in prostate cancer cells, suggesting the presence of an autocrine pathway. In this study, we show that MT1-MMP–expressing LNCaP prostate cancer cells display enhanced migration. Moreover, conditioned medium from LNCaP cells expressing both RANKL and MT1-MMP stimulates the migration of MT1-MMP–deficient C42b prostate cancer cells. This enhanced chemotaxis can be abrogated by osteoprotegerin (soluble decoy receptor of RANKL), MIK-G2 (a selective inhibitor for MT1-MMP), and PP2 (a Src inhibitor). These findings indicate that tumor-derived MT1-MMP enhances tumor cell migration through initiation of an autocrine loop requiring ectodomain shedding of membrane-bound RANKL in prostate cancer cells, and that Src is a key downstream mediator of RANKL-induced migration of prostate cancer cells. Cancer Res; 70(13); 5558–66. ©2010 AACR.
Extracellular ProteaseADAMTS9Suppresses Esophageal and Nasopharyngeal Carcinoma Tumor Formation by Inhibiting Angiogenesis
Authors: Lo Paulisally Hau Yi , Lung Hong Lok , Cheung Arthur Kwok Leung , Apte Suneel S. , Chan Kwok Wah , Kwong Fung Mei , Ko Josephine Mun Yee , Cheng Yue , Law Simon , Srivastava Gopesh , Zabarovsky Eugene R. , Tsao Sai Wah , Tang Johnny Cheuk On , Stanbridge Eric J. , Lung Maria Li
Published: June 2010

ADAMTS metalloprotease family member ADAMTS9 maps to 3p14.2 and shows significant associations with the aerodigestive tract cancers esophageal squamous cell carcinoma (ESCC) and nasopharyngeal carcinoma (NPC). However, the functional impact of ADAMTS9 on cancer development has not been explored. In this study, we evaluated the hypothesized antiangiogenic and tumor-suppressive functions of ADAMTS9 in ESCC and NPC, in stringent tumorigenicity and Matrigel plug angiogenesis assays. ADAMTS9 activation suppressed tumor formation in nude mice. Conversely, knockdown of ADAMTS9 resulted in clones reverting to the tumorigenic phenotype of parental cells. In vivo angiogenesis assays revealed a reduction in microvessel numbers in gel plugs injected with tumor-suppressive cell transfectants. Similarly, conditioned medium from cell transfectants dramatically reduced the tube-forming capacity of human umbilical vein endothelial cells. These activities were associated with a reduction in expression levels of the proangiogenic factors MMP9 and VEGFA, which were consistently reduced in ADAMTS9 transfectants derived from both cancers. Taken together, our results indicate that ADAMTS9 contributes an important function in the tumor microenvironment that acts to inhibit angiogenesis and tumor growth in both ESCC and NPC. Cancer Res; 70(13); 5567–76. ©2010 AACR.
Poly(ADP-Ribose) Polymerase Is Hyperactivated in Homologous Recombination–Defective Cells
Authors: Gottipati Ponnari , Vischioni Barbara , Schultz Niklas , Solomons Joyce , Bryant Helen E. , Djureinovic Tatjana , Issaeva Natalia , Sleeth Kate , Sharma Ricky A. , Helleday Thomas
Published: June 2010

Poly(ADP-ribose) (PAR) polymerase 1 (PARP1) is activated by DNA single-strand breaks (SSB) or at stalled replication forks to facilitate DNA repair. Inhibitors of PARP efficiently kill breast, ovarian, or prostate tumors in patients carrying hereditary mutations in the homologous recombination (HR) genes BRCA1 or BRCA2 through synthetic lethality. Here, we surprisingly show that PARP1 is hyperactivated in replicating BRCA2-defective cells. PARP1 hyperactivation is explained by the defect in HR as shRNA depletion of RAD54, RAD52, BLM, WRN, and XRCC3 proteins, which we here show are all essential for efficient HR and also caused PARP hyperactivation and correlated with an increased sensitivity to PARP inhibitors. BRCA2-defective cells were not found to have increased levels of SSBs, and PAR polymers formed in HR-defective cells do not colocalize to replication protein A or γH2AX, excluding the possibility that PARP hyperactivity is due to increased SSB repair or PARP induced at damaged replication forks. Resistance to PARP inhibitors can occur through genetic reversion in the BRCA2 gene. Here, we report that PARP inhibitor–resistant BRCA2-mutant cells revert back to normal levels of PARP activity. We speculate that the reason for the sensitivity of HR-defective cells to PARP inhibitors is related to the hyperactivated PARP1 in these cells. Furthermore, the presence of PAR polymers can be used to identify HR-defective cells that are sensitive to PARP inhibitors, which may be potential biomarkers. Cancer Res; 70(13); 5389–98. ©2010 AACR.
Exchange Protein Directly Activated by Cyclic AMP Increases Melanoma Cell Migration by a Ca2+-Dependent Mechanism
Authors: Baljinnyam Erdene , De Lorenzo Mariana S. , Xie Lai-Hua , Iwatsubo Mizuka , Chen Suzie , Goydos James S. , Nowycky Martha C. , Iwatsubo Kousaku
Published: June 2010

Melanoma has a poor prognosis due to its strong metastatic ability. Although Ca2+ plays a major role in cell migration, little is known about the role of Ca2+ in melanoma cell migration. We recently found that the exchange protein directly activated by cyclic AMP (Epac) increases melanoma cell migration via a heparan sulfate–related mechanism. In addition to this mechanism, we also found that Epac regulates melanoma cell migration by a Ca2+-dependent mechanism. An Epac agonist increased Ca2+ in several different melanoma cell lines but not in melanocytes. Ablation of Epac1 with short hairpin RNA inhibited the Epac agonist–induced Ca2+ elevation, suggesting the critical role of Epac1 in Ca2+ homeostasis in melanoma cells. Epac-induced Ca2+ elevation was negated by the inhibition of phospholipase C (PLC) and inositol triphosphate (IP3) receptor. Furthermore, Epac-induced cell migration was reduced by the inhibition of PLC or IP3 receptor. These data suggest that Epac activates Ca2+ release from the endoplasmic reticulum via the PLC/IP3 receptor pathway, and this Ca2+ elevation is involved in Epac-induced cell migration. Actin assembly was increased by Epac-induced Ca2+, suggesting the involvement of actin in Epac-induced cell migration. In human melanoma specimens, mRNA expression of Epac1 was higher in metastatic melanoma than in primary melanoma, suggesting a role for Epac1 in melanoma metastasis. In conclusion, our findings reveal that Epac is a potential target for the suppression of melanoma cell migration, and, thus, the development of metastasis. Cancer Res; 70(13); 5607–17. ©2010 AACR.
FOXO1A Is a Target for HER2-Overexpressing Breast Tumors
Authors: Wu Yanyuan , Shang Xiying , Sarkissyan Marianna , Slamon Dennis , Vadgama Jaydutt V.
Published: June 2010

Trastuzumab treatment has improved the overall survival of HER2-overexpressing breast cancer patients. However, many of these patients will eventually become resistant to treatment. The mechanisms that contribute to resistance to trastuzumab are unknown. In this study, we tested the hypothesis that targeting of the FKHR transcription factor FOXO1A in HER2-overexpressing breast tumor cells can overcome the trastuzumab resistance in vitro. We have shown that overexpression of HER2 leads to activation of phosphatidylinositol 3-kinase (PI3K)/Akt pathway and subsequent inactivation of FOXO1A in HER2-overexpressing breast cancer cells SKBR3, BT474, and MCF7-HER2. In wild-type SKBR3 and BT474 cells, trastuzumab downregulates active Akt and increases FOXO1A expression that leads to increase in p27kip1 and decrease in cyclin D1 and finally inhibits cell proliferation. In contrast, the effect of trastuzumab was eliminated by the reduction of FOXO1A in HER2-overexpressing cells with constitutively active Akt1 (SKBR3/AA28 and BT474/AA9). The downregulation of FOXO1A resulted in nuclear export of p27kip1. Blocking the constitutively active Akt by a specific Akt/protein kinase B signaling inhibitor-2 (API-2) significantly increased FOXO1A expression and rendered the cells more responsive to trastuzumab-induced growth inhibition. Reactivation of FOXO1A by stable or transient transfection also restored the growth-inhibitory effects of trastuzumab in SKBR3/AA28, BT474/AA9, and MCF7-HER2 cells. Knocking down FOXO1A by small interfering RNA resulted in reducing trastuzumab-induced growth inhibition. In summary, trastuzumab can inhibit proliferation of HER2-overexpressing breast cancer cells by reactivating FOXO1A through inhibition of the PI3K/Akt pathway. FOXO1A may therefore serve as a target for HER2-overexpressing breast tumors. Cancer Res; 70(13); 5475–85. ©2010 AACR.
Src Family Kinase Inhibitor Saracatinib (AZD0530) Impairs Oxaliplatin Uptake in Colorectal Cancer Cells and Blocks Organic Cation Transporters
Authors: Morrow Christopher J. , Ghattas Mohammad , Smith Christopher , Bönisch Heinz , Bryce Richard A. , Hickinson D. Mark , Green Tim P. , Dive Caroline
Published: June 2010

Elevated Src family kinase (SFK) activity is associated with tumor invasion and metastasis. The SFK inhibitor saracatinib (AZD0530) is currently in phase II trials in patients including those with colorectal cancer (CRC), where links between SFK activity and poor prognosis are particularly striking. Saracatinib is likely to be used clinically in combination regimens, specifically with 5-fluorouracil (5-FU) and oxaliplatin, in CRC. The aim of this study was to determine the effect of saracatinib on oxaliplatin and 5-FU efficacy in CRC cells. Saracatinib did not modulate 5-FU efficacy but antagonized oxaliplatin in a schedule-specific manner through reduced oxaliplatin uptake via an SFK-independent mechanism. Saracatinib resembles the pharmacophore of known organic cation transporter (OCT) inhibitors and reduced oxaliplatin efficacy maximally in cells overexpressing OCT2. These data suggest that oxaliplatin uptake in CRC is attenuated by saracatinib via inhibition of OCT2, a potential consideration for the clinical development of this SFK inhibitor. Cancer Res; 70(14); 5931–41. ©2010 AACR.
Matrix Metalloproteinase-1 Expression Can Be Upregulated through Mitogen-Activated Protein Kinase Pathway under the Influence of Human Epidermal Growth Factor Receptor 2 Synergized with Estrogen Receptor
Authors: Jung Hae Hyun , Park Yeon Hee , Jun Hyun Jung , Kong Jeehyun , Kim Jeong Hoon , Kim Jung A , Yun Jina , Sun Jong Mu , Won Young Woong , Lee Soohyeon , Kim Seung Tae , Ahn Jin Seok , Im Young-Hyuck
Published: June 2010

In our previous work, Ets-1 upregulates human epidermal growth factor receptor 2 (HER2) induced matrix metalloproteinase 1 (MMP-1) expression. Based on the above knowledge and result, we hypothesized that estrogen receptor (ER) and its signaling pathway may affect MMP-1 expression under the influence of HER2. In addition, we investigated how the HER2 pathway cross-talk with the ER signaling pathway in genomic and nongenomic action of ER using reverse transcription-PCR, Western blot analysis, and ELISA assay. The results showed that ER-α expression increased MMP-1 expression under the presence of HER2. These upregulatory effects were mediated mainly by mitogen-activated protein kinase pathway and were reversed by downregulation of HER2 and/or ER. Activator protein DNA binding activity was involved in the MMP-1 expression. In summary, our results showed that ER can upregulate MMP-1 expression under the influence of HER2 in MCF-7 cells. In addition, this upregulatory effect was found to be mediated by mitogen-activated protein kinase pathway. MMP-1 might be an assigned target in interaction between ER and HER2. Mol Cancer Res; 8(7); 1037–47. ©2010 AACR.
Quercetin Potentiates UVB-Induced c-Fos Expression: Implications for Its Use as a Chemopreventive Agent
Authors: Olson Erik R. , Melton Tania , Dickinson Sally E. , Dong Zigang , Alberts David S. , Bowden G. Tim
Published: June 2010

Quercetin (Qu) is currently being investigated as a chemopreventive agent for several cancers, including nonmelanoma skin cancer induced by UV light. We previously reported that Qu degradation has important consequences on signaling and cell biology. In the current study, we report that Qu induces c-Fos mRNA and protein expression through activation of p38 and cAMP-responsive element binding protein (CREB), and Qu potentiates UVB-induced c-Fos expression. Inclusion of ascorbic acid (AA) in cell culture medium stabilizes Qu and completely prevents both Qu- and UVB-induced p38 and CREB activation, leading to a blockade of c-fos gene expression through reduced CREB/cAMP-responsive element binding. AA stabilizes c-Fos mRNA, increasing steady-state levels even when c-fos gene expression is suppressed, but this has no effect on c-Fos protein levels in either mock- or UVB-irradiated cells. We report that Qu blocks mammalian target of rapamycin signaling and inhibits c-Fos protein expression directly through this mechanism because cotreatment with Qu and AA resulted in the complete suppression of UVB-induced c-Fos protein expression even in the presence of significantly increased mRNA levels. We further confirmed that this was not due to increased protein turnover because inhibition of proteasome activity with MG-132 did not raise c-Fos protein levels in Qu+AA-treated cells. Together, these data indicate that although Qu has been reported to have some beneficial properties as a chemopreventive agent, it is also capable of inducing c-fos expression, a cellular event important for the promotion phase of tumor development, if it is not stabilized. Cancer Prev Res; 3(7); 876–84. ©2010 AACR.
Meat Consumption, Nonsteroidal Anti-Inflammatory Drug Use, and Mortality among Colorectal Cancer Patients in the California Teachers Study
Authors: Zell Jason A. , Ziogas Argyrios , Bernstein Leslie , Clarke Christina A. , Deapen Dennis , Largent Joan A. , Neuhausen Susan L. , Stram Daniel O. , Ursin Giske , Anton-Culver Hoda
Published: June 2010

A low-meat diet and regular use of nonsteroidal anti-inflammatory drugs (NSAID) have been associated with decreased mortality among colorectal cancer (CRC) patients. Here, we investigated the association between prediagnosis usual meat consumption and CRC-specific mortality, and whether meat consumption modifies the previously noted association between NSAID use and CRC-specific mortality among women in the California Teachers Study cohort. Women joining the California Teachers Study in 1995-1996 without prior CRC diagnosis, diagnosed with incident CRC during follow-up through December 2007, were eligible for inclusion. Meat intake (frequency and serving size) and NSAID use (aspirin or ibuprofen use) were ascertained via self-administered questionnaires before diagnosis. Vital status and cause of death were determined by linkage with mortality files. Multivariable Cox proportional hazards regression models were used to estimate hazard ratios for death and 95% confidence intervals. Prediagnosis meat consumption was not associated with CRC-specific mortality among 704 CRC patients (and 201 CRC-specific deaths), comparing patients in the lowest consumption tertile (0-5.4 medium-sized servings/wk) to those in the higher consumption tertiles. Regular NSAID use (1-3 times/wk, 4-6 times/wk, daily) versus none was associated with decreased CRC-specific mortality among patients in the lowest meat consumption tertile (hazard ratio, 0.22; 95% CI, 0.06-0.82), but not among patients in the higher meat intake tertiles. The previously observed mortality risk reduction among female CRC patients associated with regular NSAID use was restricted to patients who reported low meat intake before diagnosis. These findings have implications for CRC survivorship and tertiary CRC prevention. Cancer Prev Res; 3(7); 865–75. ©2010 AACR.
Impact of Dementia on Hospitalization
Authors: Guijarro Ricardo , San Román Carlos M. , Gómez-Huelgas Ricardo , Villalobos Aurora , Martín Magdalena , Guil María , Martínez-González Miguel-Ángel , Toledo Juan B.
Published: June 2010

Background: The purpose of this study was to analyze and determine the prevalence and clinical characteristics of hospitalized dementia patients compared with nondemented patients. Methods: We examined hospital discharge database records dated 1998–2003 from public hospitals in Andalusia, Spain. We used ICD-9-CM codes to identify patients with dementia. The variables examined included age, length of stay, discharge diagnosis, diagnostic-related groups, and mortality of both dementia and nondementia patients over 65 years of age. Results: A diagnosis of dementia was documented for 40,482 cases. The prevalence of dementia increased from 3.43% to 4.64% between 1998 and 2003 and was higher among older patients and women. Dementia was the reason for admission in 5.6% of cases. Medical reasons constituted 82.4% of admittances. Dementia patients had hip surgery more frequently than patients without dementia, and other procedures (orthopedic surgery, cataracts, or hernia repair) were less frequent (p < 0.001). The mean duration of the hospital stay was longer (13.4 vs. 10.7 days) and the intra-hospital mortality rate was greater (19.3% vs. 8.7%) for patients with dementia compared to those without dementia. Dementia was an independent predictor of mortality (OR 1.77; 95% CI 1.72–1.82). Conclusions: Dementia is increasing among hospitalized patients. Dementia patients have different reasons for hospitalization and higher mortality. It is necessary to identify these differences and to improve the hospital care of dementia patients.
Mobile Phone Use and the Risk for Malignant Brain Tumors: A Case-Control Study on Deceased Cases and Controls
Authors: Hardell Lennart , Carlberg Michael , Hansson Mild Kjell
Published: June 2010

We investigated the use of mobile or cordless phones and the risk for malignant brain tumors in a group of deceased cases. Most previous studies have either left out deceased cases of brain tumors or matched them to living controls and therefore a study matching deceased cases to deceased controls is warranted. Recall error is one issue since it has been claimed that increased risks reported in some studies could be due to cases blaming mobile phones as a cause of the disease. This should be of less importance for deceased cases and if cancer controls are used. In this study brain tumor cases aged 20–80 years diagnosed during 1997–2003 that had died before inclusion in our previous studies on the same topic were included. Two control groups were used: one with controls that had died from another type of cancer than brain tumor and one with controls that had died from other diseases. Exposure was assessed by a questionnaire sent to the next-of-kin for both cases and controls. Replies were obtained for 346 (75%) cases, 343 (74%) cancer controls and 276 (60%) controls with other diseases. Use of mobile phones gave an increased risk, highest in the >10 years’ latency group yielding odds ratio (OR) = 2.4, and 95% confidence interval (CI) = 1.4–4.1. The risk increased with cumulative number of lifetime hours for use, and was highest in the >2,000 h group (OR = 3.4, 95% CI = 1.6–7.1). No clear association was found for use of cordless phones, although OR = 1.7, 95% CI = 0.8–3.4 was found in the group with >2,000 h of cumulative use. This investigation confirmed our previous results of an association between mobile phone use and malignant brain tumors.
Amoeba/Amoebal Symbiont Genetic Transfers: Lessons from Giant Virus Neighbours
Authors: Thomas Vincent , Greub Gilbert
Published: June 2010

Free-living amoebae serve as hosts for a variety of amoebae-resisting microorganisms, including giant viruses and certain bacteria. The latter include symbiotic bacteria as well as bacteria exhibiting a pathogenic phenotype towards amoebae. Amoebae-resisting bacteria have been shown to be widespread in water and to use the amoebae as a reservoir, a replication niche, a protective armour as well as a training ground to select virulence traits allowing survival in the face of microbicidal effects of macrophages, the first line of defense against invading pathogens. More importantly, amoebae play a significant role as a melting pot for genetic exchanges. These ecological and evolutionary roles of amoebae might also be at play for giant viruses and knowledge derived from the study of amoebae-resisting bacteria is useful for the study and understanding of interactions between amoebae and giant viruses. This is especially important since some genes have spread in all domains of life and the exponential availability of eukaryotic genomes and metagenomic sequences will allow researchers to explore these genetic exchanges in a more comprehensive way, thus completely changing our perception of the evolutionary history of organisms. Thus, a large part of this review is dedicated to report current known gene exchanges between the different amoebae-resisting organisms and between amoebae and the internalized bacteria.
A Dilemma for Viruses and Giant Viruses: Which Endocytic Pathway to Use to Enter Cells?
Authors: Ghigo Eric
Published: June 2010

Viruses must enter host cells to deliver their genetic material and accessory proteins. Endocytosis offers to viruses the opportunity to enter host cells. However, endocytosis is a complex phenomenon that includes different mechanisms, clathrin-mediated endocytosis, caveolin-mediated endocytosis, macropinocytosis, and phagocytosis. Here, I describe the ways used by different viruses to exploit these endocytic pathways.
Origin and Evolution of Eukaryotic Large Nucleo-Cytoplasmic DNA Viruses
Authors: Koonin Eugene V. , Yutin Natalya
Published: June 2010

Background/Aims: The nucleo-cytoplasmic large DNA viruses (NCLDV) constitute an apparently monophyletic group that consists of 6 families of viruses infecting a broad variety of eukaryotes. A comprehensive genome comparison and maximum-likelihood reconstruction of NCLDV evolution reveal a set of approximately 50 conserved genes that can be tentatively mapped to the genome of the common ancestor of this class of eukaryotic viruses. We address the origins and evolution of NCLDV. Results: Phylogenetic analysis indicates that some of the major clades of NCLDV infect diverse animals and protists, suggestive of early radiation of the NCLDV, possibly concomitant with eukaryogenesis. The core NCLDV genes seem to have originated from different sources including homologous genes of bacteriophages, bacteria and eukaryotes. These observations are compatible with a scenario of the origin of the NCLDV at an early stage of the evolution of eukaryotes through extensive mixing of genes from widely different genomes. Conclusions: The common ancestor of the NCLDV probably evolved from a bacteriophage as a result of recruitment of numerous eukaryotic and some bacterial genes, and concomitant loss of the majority of phage genes except for a small core of genes coding for proteins essential for virus genome replication and virion formation.
Inside the Lifestyle of the Virophage
Authors: Desnues C. , Raoult D.
Published: June 2010

Objective(s): We sought to better characterize Sputnik, the first isolated virophage, and to analyze its parasitic lifestyle during co-infection with Marseillevirus (a new giant virus) in Acanthamoeba castellanii. Methods: A combination of electron microscopy, immunofluorescence microscopy, and real-time PCR was used to characterize the kinetics of the viral replication cycle. RT-PCR was performed to detect RNAs inside the Sputnik virions. Results: Sputnik is a new viral entity carrying an almost complete ready-to-use set of viral RNAs (20 out of 21). Sputnik does not replicate with Marseillevirus but delays its replication cycle. While Marseillevirus is successfully internalized by A. castellanii following co-infections with Mamavirus and Sputnik, it does not initiate a replication cycle. In contrast, both Marseillevirus and Mamavirus can replicate in the amoeba in case of co-infection, but the development of one is exclusive from the other inside a single amoeba cell. Conclusions: This work provides new insight into the Sputnik replication cycle with another giant virus and confirms that Sputnik is a virophage. It shows new dimensions of the interactions existing among giant viruses.
Advances in Mimivirus Pathogenicity
Authors: Vincent Agnès , La Scola Bernard , Papazian Laurent
Published: June 2010

Viral diseases in the clinical setting have been extensively investigated. Viruses are now considered as potentially responsible for nosocomial infections, especially in intensive care unit (ICU) patients. Mimivirus is the largest virus known to date. Recent studies have suggested that Mimivirus could be responsible for both community-acquired and nosocomial pneumonia. These studies were mainly based on serologic diagnosis, which showed patients with community-acquired pneumonia have more antibodies to Mimivirus than healthy controls. Serologic evidence of Mimivirus pneumonia was also found in mechanically ventilated ICU patients. In a matched-cohort study in which ICU patients with serologic evidence of Mimivirus pneumonia were matched to ICU patients remaining seronegative for Mimivirus, positive serology was associated with an increased duration of both mechanical ventilation and ICU stay. Identification by PCR techniques remains difficult, probably because of the high level of polymorphism of nucleotide sequences of giant viruses. More studies are needed to confirm the clinical impact of Mimivirus in humans.
Classification and Determination of Possible Origins of ORFans through Analysis of Nucleocytoplasmic Large DNA Viruses
Authors: Boyer Mickaël , Gimenez Gregory , Suzan-Monti Marie , Raoult Didier
Published: June 2010

Objective: An important proportion of coding sequences in genomes, notably in viruses, do not match any sequences in databases and are assigned as ORFan sequences. Nucleocytoplasmic large DNA viruses (NCLDVs) harbor great numbers of ORFs with a high number consisting of ORFans. Thus, we decided to decipher the nature of ORFans in the NCLDVs. Methods: A genome-wide study was carried out to estimate the ORFan proportion in NCLDV genomes and to analyze their general features compared with non-ORFan. Results: The ORFan percentages comprised between 2.8 and 75.2% of the ORF content according to the virus lineage. We propose to classify ORFans in four categories according to their possible match with metagenomic sequences and their prevalence at different taxonomic ranks. Our results indicate that NCLDV ORFans have overall similar features with non-ORFans, except they are shorter. Conclusions: An ORFan classification scheme was proposed to decipher their origin and evolution. Most ORFans were likely labeled ORFan owing to the gap of knowledge of the sequence space. ORFans might be true functional genes with likely the same expression potential as non-ORFan genes. Part of them may also correspond to new genes formed de novo through the diverse mechanisms of gene evolution.
Amoebae as Genitors and Reservoirs of Giant Viruses
Authors: Raoult Didier , Boyer Mickael
Published: June 2010

Amoebae are unicellular phagocytes that feed on microorganisms in their environment. Some amoebae have the largest genome size currently known on earth. They phagocytose any inert particle larger than 0.5 µm. Phagocytic amoebae can harbor different bacteria, fungi and giant viruses within the same cell. There is evidence of lateral gene transfer between the amoeba and its microbiological hosts. There is also evidence of gene exchange between viruses and bacteria hosted in amoebae. Moreover, there is evidence of gene transfer between viruses, such as Mimivirus and the virophage. As a consequence, viruses and intracellular bacteria living in amoebae have a sympatric lifestyle and large chimeric genome repertoires. We conclude that phagocytic protists continuously generate new species with chimeric repertoires that may succeed later if adapted to the environmental conditions and selected in a specific niche.
In vitro and in vivo Cytogenetic Effects of Recombinant Human Erythropoietin on the Frequency of Sister Chromatid Exchanges Alone or in Combination with Mitomycin C
Authors: Digkas E.N. , Chrisafi S. , Passadaki T. , Tsalkidis A. , Hatzimichail A. , Vargemezis V. , Lialiaris T.S.
Published: June 2010

Background: Erythropoietin (EPO) is a glycoprotein which has a main property, erythropoiesis, but its range of action in the human body is very wide. It has been suggested that EPO acts cytoprotectively for many cell lines against many toxic causes in vitro and in vivo. Our aim was to study the action of EPO on DNA of two cell types, human lymphocytes in vitro and on P388 ascites tumor cells inoculated in BDF1 mice in the presence and absence of the genotoxic agent mitomycin C (MMC). Method: The sister chromatid exchange (SCE) assay was used as it is a very sensitive, simple and rapid method for detecting DNA damage. Proliferation rate indices (PRI) and mitotic indices (MI) were also counted. Results: EPO did not alter the SCE level when it acted alone on both cell lines. MMC as a potent genotoxic agent increased SCE levels in vitro and in vivo. EPO used in combination with MMC significantly decreased SCE levels and increased PRI and MI values induced by MMC alone both in vitro and in vivo. Conclusions: EPO acts protectively against the genotoxic potential of MMC, and this action may have clinical implications.
Mediterranean Dietary Pattern Is Associated With Improved Cardiac Autonomic Function Among Middle-Aged Men
Authors: Dai Jun , Lampert Rachel , Wilson Peter W. , Goldberg Jack , Ziegler Thomas R. , Vaccarino Viola
Published: June 2010

Background—

Reduced heart rate variability (HRV), a measure of cardiac autonomic dysfunction, is a risk factor for coronary artery disease. Diet can influence HRV, but this association may be confounded by genetic and environmental factors.

Methods and Results—

We administered the Willett Food Frequency Questionnaire to 276 middle-aged male twins. We derived a score measuring the extent to which an individual's diet conformed to the Mediterranean diet following a published algorithm. The higher the score, the greater the similarity to the Mediterranean diet. All twins underwent 24-hour ambulatory ECG recording. Time and frequency domain measures of HRV were calculated. Mixed-effects regression was used to partition the association into between- and within-twin pair differences. After adjusting for energy intake, other nutritional factors, shared genes, and common environment, a 1-unit higher score was significantly associated with 3.9% to 13% higher time and frequency domain HRV parameters. Further controlling for known cardiovascular risk factors and use of fish oil supplements and medications did not substantially change the estimates.

Conclusions—

The Mediterranean dietary pattern is associated with higher HRV.
Paskuwatti’s Ritual: Remedy for Impotence or Antidote to Homosexuality?
Authors: Miller Jared L.
Published: June 2010

The Hittite composition known as Paskuwatti’s Ritual (CTH 406) has long been understood as a rite designed to help a man to overcome sexual impotence. The present paper, in contrast, attempts to demonstrate that it is more convincingly interpreted as an antidote to homosexual behavior, apparently seeking to cure its patient of his proclivity for passive sexual acquiescence and to replace it with an inclination toward normative male, i.e. penetrative behavior.
Analysing Greek-Turkish disaster-related cooperation: A disaster diplomacy perspective
Authors: Ganapati N. Emel , Kelman Ilan , Koukis Theodore
Published: June 2010

This article contributes to the disaster diplomacy literature in examining the conditions under which disasters can lead to long-term disaster-related collaboration (e.g. in disaster response, recovery or risk reduction) at both governmental and non-governmental level among states in conflict. In particular, the article focuses on the role of the 1999 earthquakes in enhancing such collaboration between Greece and Turkey over the past decade. While acknowledging the diversity and complexity of disaster diplomacy situations, the article suggests that disasters can lead to long-term disaster-related cooperation among states in conflict when: (1) one party providing disaster relief to another party is followed by a similar reciprocal gesture (i.e. tit-for-tat diplomacy); (2) there is a realization and acceptance that neighbours should come to each other’s assistance in times of disaster; and (3) there is an enabling broader context (e.g. a rapprochement process) conducive to sustaining the long-term cooperation.
Democratic accountability of international organizations: Parliamentary control within the Council of Europe and the OSCE and the prospects for the United Nations
Authors: Habegger Beat
Published: June 2010

Many international organizations, including the United Nations, are accused of lacking democratic accountability. A variety of proposals have been made to close the gap between their extended influence and the lack of effective controlling mechanisms to prevent abuses of power. This article focuses on one specific proposal: the establishment of parliamentary assemblies. Based on the experiences of the Council of Europe and the Organization for Security and Cooperation in Europe, it presents the factors that enable such assemblies to exercise parliamentary control and influence towards intergovernmental decision-making bodies.The article shows how a parliamentary dimension can be introduced conceptually into the debate on the democratic accountability of international organizations and how these insights support the reflections on a United Nations parliamentary assembly.
Targeting Nuclear Programs in War and Peace: A Quantitative Empirical Analysis, 1941-2000
Authors: Fuhrmann Matthew , Kreps Sarah E.
Published: June 2010

When do states attack or consider attacking nuclear infrastructure in nonnuclear weapons states? Despite the importance of this question, relatively little scholarly research has considered when and why countries target nuclear programs. The authors argue that states are likely to attack or consider attacking nuclear facilities when they are highly threatened by a particular country’s acquisition of nuclear weapons. Three factors increase the salience of the proliferation threat: (1) prior violent militarized conflict; (2) the presence of a highly autocratic proliferator; and (3) divergent foreign policy interests. The authors test these propositions using statistical analysis and a new data set on all instances when countries have struck or seriously considered striking other states’ nuclear infrastructure between 1941 and 2000. The findings lend support for the theory and very little support for the alternative explanations. States are not deterred from attacking nuclear programs by the prospect of a military retaliation and concerns about international condemnation do not appear to influence the willingness to strike. Ultimately, states are willing to accept substantial costs in attacking if they believe that a particular country’s acquisition of nuclear weapons poses a significant threat to their security.
Review Essay: Is Homo Economics Extinct?
Authors: Sassower Raphael
Published: June 2010

The classical view of “rational man” as the unit of analysis for economic behavior and marketplace exchange has been changed by the late twentieth century with the help of behavioral economics that considers predictable irrationality as a normal mode of behavior. Instead of revising neoclassical economics to fit contemporary economic crises, it is recommended to follow Adam Smith's original concerns for the social setting of individual behavior and to treat economic crises with pragmatic flexibility rather than with dogmatic ideology.
Flavivirus Encephalitis
Authors: Kimura T. , Sasaki M. , Okumura M. , Kim E. , Sawa H.
Published: June 2010

Encephalitic flaviviruses are important arthropod-borne pathogens of humans and other animals. In particular, the recent emergence of the West Nile virus (WNV) and Japanese encephalitis virus (JEV) in new geographic areas has caused a considerable public health alert and international concern. Among the experimental in vivo models of WNV and JEV infection, mice and other laboratory rodents are the most thoroughly studied and well-characterized systems, having provided data that are important for understanding the infectious process in humans. Macaca monkeys have also been used as a model for WNV and JEV infection, mainly for the evaluation of vaccine efficacy, although a limited number of published studies have addressed pathomorphology. These animal models demonstrate the development of encephalitis with many similarities to the human disease; however, the histological events that occur during infection, especially in peripheral tissues, have not been fully characterized.
REVIEW PAPER
Authors: Steele K. E. , Twenhafel N. A.
Published: June 2010

The encephalitides caused by Venezuelan (VEEV), eastern (EEEV), and western (WEEV) equine encephalitis viruses are important natural diseases of horses and humans and potential agents of biowarfare or bioterrorism. No licensed vaccines or specific therapies exist to prevent or treat human infections with VEEV, EEEV, or WEEV. Well-characterized animal models are needed to support the development of such medical countermeasures under the United States Food and Drug Administration’s “Animal Rule.” This review focuses on the pathological features and pathogenetic mechanisms of these alphaviral encephalitides in animal models, with an emphasis on aerosol infections. Infection of mice, nonhuman primates, and other species with VEEV, EEEV, and WEEV causes encephalitis and often death. There is great variability in the specific manifestations of disease in the different models, however. Many aspects of the disease in animal models and in humans remain to be characterized using modern methods. Especially needed is a better understanding of the fundamental mechanisms involved in 3 key phases of the pathogenesis of alphavirus encephalitis. These are the early extraneural phase, the process of neuroinvasion itself, and virus and host factors related to neurovirulence. A greater understanding of these aspects could provide avenues for the development of medical countermeasures and better establish suitable animal models of alphavirus encephalitis for testing them under the Animal Rule.
An early 18th-century case description of cluster headache
Authors: Arkink EB , van Buchem MA , Haan J , Ferrari MD , Kruit MC
Published: June 2010

We present a previously unreported early 18th-century description of cluster headache by the English antiquary Abraham de la Pryme (1671–1704) initially attributed to hydrophobia (rabies). We will also give a short overview of other descriptions of cluster and cluster-like headache in historical literature.
Preclinical pharmacological profile of the selective 5-HT1Freceptor agonist lasmiditan
Authors: Nelson David L , Phebus Lee A , Johnson Kirk W , Wainscott David B , Cohen Marlene L , Calligaro David O , Xu Yao-Chang
Published: June 2010

Introduction: Lasmiditan (also known as COL-144 and LY573144; 2,4,6-trifluoro- N-[6-[(1-methylpiperidin-4-yl)carbonyl]pyridin-2yl]benzamide) is a high-affinity, highly selective serotonin (5-HT) 5-HT1Freceptor agonist.

Results: In vitro binding studies show a Kivalue of 2.21 nM at the 5-HT1Freceptor, compared with Kivalues of 1043 nM and 1357 nM at the 5-HT1Band 5-HT1Dreceptors, respectively, a selectivity ratio greater than 470-fold. Lasmiditan showed higher selectivity for the 5-HT1Freceptor relative to other 5-HT1receptor subtypes than the first generation 5-HT1Freceptor agonist LY334370. Unlike the 5-HT1B/1Dreceptor agonist sumatriptan, lasmiditan did not contract rabbit saphenous vein rings, a surrogate assay for human coronary artery constriction, at concentrations up to 100 µM. In two rodent models of migraine, oral administration of lasmiditan potently inhibited markers associated with electrical stimulation of the trigeminal ganglion (dural plasma protein extravasation, and induction of the immediate early gene c-Fos in the trigeminal nucleus caudalis).

Conclusions: Lasmiditan presents a unique pyridinoyl-piperidine scaffold not found in any other antimigraine class. Its chemical structure and pharmacological profile clearly distinguish it from the triptans. The potency and selectivity of lasmiditan make it ideally suited to definitively test the involvement of 5-HT1Freceptors in migraine headache therapy.
Acute treatment of migraine with the selective 5-HT1F receptor agonist lasmiditan – A randomised proof-of-concept trial
Authors: Ferrari Michel D , Färkkilä Markus , Reuter Uwe , Pilgrim Alison , Davis Charles , Krauss Martin , Diener Hans-Christoph ,   for the European COL-144 Investigators
Published: June 2010

Introduction: Lasmiditan (COL-144; LY573144) is a novel, highly selective and potent agonist at 5-HT1F receptors that lacks vasoconstrictor activity. Preclinical and early clinical experiments predict acute antimigraine efficacy of COL-144 that is mediated through a non-vascular, primarily neural, mechanism.

Subjects and methods: In a randomised, multicentre, placebo-controlled, double-blind, group-sequential, adaptive treatment-assignment, proof-of-concept and dose-finding study, we treated 130 subjects in-hospital during a migraine attack. Subjects were allocated to an intravenous dose level of lasmiditan or placebo in small cohorts. The starting dose was 2.5 mg. Subsequent doses were adjusted, up or down, according to the safety and efficacy seen in the preceding cohort. The primary outcome measure was headache response defined as improvement from moderate or severe headache at baseline to mild or no headache at 2 h post-dose. The study was designed to explore the overall dose response relationship but was not powered to differentiate individual doses from placebo, nor to detect effect differences for other migraine symptoms.

Results: Forty-two subjects received placebo and 88 received lasmiditan in doses of 2.5–45 mg. Subjects were observed in the clinic for 4 h after treatment and used a diary card to record symptoms and adverse events for up to 24 h. The study was terminated when the 20 mg dose met predefined efficacy stopping rules. Of subjects treated in the 10, 20, 30 and 45 mg lasmiditan dose groups, 54–75% showed a 2 h headache response, compared to 45% in the placebo group ( P = 0.0126 for the linear association between response rates and dose levels). Patient global impression at 2 h and lack of need for rescue medication also showed statistically significant linear correlations with dose. Lasmiditan was generally well tolerated. Adverse events were reported by 65% of subjects on lasmiditan and by 43% on placebo and were generally mild. Dizziness, paresthesia and sensations of heaviness (usually limb) were more common on lasmiditan.

Conclusions: At intravenous doses of 20 mg and higher, lasmiditan proved effective in the acute treatment of migraine. Further studies to assess the optimal oral dose and full efficacy and tolerability profile are under way. The non-vascular, neural mechanism of action of lasmiditan may offer an alternative means to treat migraine especially in patients who have contra-indications for agents with vasoconstrictor activity. The clinicaltrials.gov identifier for this study is NCT00384774.