bluuebunny/crossref_metadata_2025 huggingface dataset:
The colonization ratio: a measure of pathogen invasiveness and host resistance in Verticillium wilt of alfalfa
Authors: Newcombe A. G. , Papadopoulos Y. A. , Robb Jane , Christie B. R.
Published: October 2007

Seeds of four alfalfa cultivars were grown and the plants cloned by rooting cuttings. Individual plants were root-dip inoculated with Verticillium albo-atrum Reinke and Berthold, and scored for relative pathogen resistance by symptom evaluation and reisolation of the fungus during a 7-month period. Nine individual plants were selected: two were resistant to colonization and symptom expression, two remained colonized but were resistant to symptom expression, and five were susceptible. Disease-free cuttings of each selected plant were infused with a mixture of conidia and red vinyl particles which mark primary spore trapping sites. At 168-h postinoculation the colonization ratio was determined for each sample. The colonization ratio is a reliable quantitative measure of invasiveness which is calculated by scoring a section of tissue for the number of secondary colonization sites established per primary trapping site in a given unit of time. The colonization ratio values of the nine selected plants were correlated with the level of disease resistance as determined by two methods: (i) symptom evaluation and pathogen resiolation, and (ii) genetic S1 analysis. The colonization ratio, a new measure of fungal invasiveness and host resistance, might be useful in the breeding and assessment of alfalfa cultivars with resistance to Verticillium wilt.
Ultrastructure of stromatal anamorphs in the Sclerotiniaceae
Authors: Kohn Linda M. , Grenville Douglas J.
Published: October 2007

As part of comparative anatomical, histochemical, and ultrastructural studies of stromata in the Sclerotiniaceae, mature stromata produced in vitro by 11 species representing six genera and one form-genus were examined using transmission electron microscopy. Sclerotial-stromatal taxa were Sclerotinia sclerotiorum, S. trifoliorum, S. minor, Sclerotium cepivorum, Botrytis cinerea, B. porri, Monilinia fructicola, and Myriosclerotinia borealis. Substratal-stromatal taxa were Sclerotinia homoeo-carpa, Rutstroemia sydowiana, and Lambertella subrenispora. Three types of rind were observed: a living cellular rind, a dead cellular rind, and a stromatal rind. Sclerotial species were distinguished from stromatal species not only by the rind type, but also by the confluent extracellular matrix around cortical and medullary cell walls. Presence of lacunae in this matrix distinguished Sclerotinia spp. and M. borealis from Botrytis spp. and Monilinia fructicola. Rind, cortical, and medullary cells contained abundant storage vacuoles in most taxa. The distribution and proportion of organelles to storage vacuoles differed among taxa. Plugged septal pores with associated Woronin bodies were similar among the taxa where they were observed. Sclerotia of Sclerotium cepivorum, which has no known teleomorph, are ultrastructurally most like sclerotia of Sclerotinia or Botrytis anamorphs of Botryotinia species. Substratal stromata of S. homoeocarpa showed unusually complex cellular organization. Sclerotial stromata of M. fructicola contained unusual storage vacuoles with heterogeneous contents.
Flagellar apparatus of male gametes and other aspects of gamete and zygote ultrastructure in Prasiola and Rosenvingiella (Chlorophyta, Prasiolales) from British Columbia
Authors: O'Kelly Charles J. , Garbary David J. , Floyd Gary L.
Published: October 2007

We examined gametes, syngamy, and zygote formation in Prasiola meridionalis Setchell & Gardner and Rosenvingiella constricta (S. & G.) Silva with the transmission electron microscope. We compare the structures we observed with those of the Atlantic species P. stipitata Suhr and describe some previously unreported features. The sperm possessed a number of unusual structural elements in the flagellar apparatus and elsewhere, in particular a 9 + 1 microtubule configuration in the flagellar axoneme. The flagellar apparatus exhibited 180° rotational symmetry and counterclockwise absolute orientation of major components. At syngamy, the flagellar apparatus changed little in composition and arrangement, suggesting that it has a minimal, if any, function in syngamy or the planozygote. Karyogamy occurred within a few hours of plasmogamy, after settlement of the planozygote and concurrently with wall formation. The three species, P. meridionalis, P. stipitata, and R. constricta, are very similar in gamete structure, which indicates that the genera Prasiola and Rosenvingiella are close relatives. The flagellar apparatus features provide evidence supporting recognition of the order Prasiolales, which may be assigned to either the class Ulvophyceae or the class Pleurastrophyceae.
Restriction fragment length polymorphisms of the mitochondrial DNA of Phytophthora megasperma isolated from soybean, alfalfa, and fruit trees
Authors: Förster Helga , Kinscherf T. G. , Leong S. A. , Maxwell Douglas P.
Published: October 2007

The mtDNAs from 24 small oogonial soybean-specific Phytophthora megasperma f.sp. glycinea (Pmg) isolates, 10 small oogonial alfalfa-specific P. megasperma f.sp. medicaginis (Pmm) isolates, 2 small oogonial nonhost-specific P. megasperma isolates from apple, 6 large oogonial nonhost-specific P. megasperma isolates from alfalfa or cherry, and 1 high-temperature P. megasperma isolate from alfalfa were investigated by restriction fragment length polymorphism analysis. Restriction patterns of the mtDNAs obtained by digestion with six restriction endonucleases showed limited variation within isolates of Pmg and Pmm. However, restriction patterns of Pmg and Pmm mtDNAs were very different. All nonhost-specific isolates of P. megasperma and the high-temperature isolate had restriction patterns that differed from each other and from those of Pmg and Pmm. The restriction patterns of the nuclear DNAs also distinguished these isolates from Pmg and Pmm. Phylogenetic analyses indicate that Pmg and Pmm are genetically distinct subgroups that represent biological species of P. megasperma and that several additional subgroups, possibly biological species, are present in the P. megasperma complex.
Taxonomy and ecology of Cylindrodendrum album var. paralion var.nov.
Authors: Summerbell Richard C. , Seifert Keith A. , Mackinnon J. Andrew
Published: October 2007

Cylindrodendrum album var. paralion var.nov. is described. The organism was isolated from decaying Fucus distichus thalli collected in British Columbia, Canada. It differs from the type variety by its strong vinaceous pigmentation and its ability to grow in the presence of 7% sodium chloride in Sabouraud agar in vitro. The organism rapidly colonizes decaying F. distichus thalli under simulated marine conditions in vitro and shows extensive growth and sporulation in the presence of other microfungi commonly isolated from the same substrate.
A FINITE ELEMENT APPROACH USING EQUIVALENT PLANE RECTANGULAR TRUSS ELEMENT IN NONLINEAR ANALYSIS OF REINFORCED CONCRETE STRUCTURES
Authors: WU FANGBO , DING XIANLI , HE SHAOYAO
Published: October 2007

A new finite element approach described in this paper uses an equivalent plane rectangular truss element to replace the traditional plane rectangular element in the nonlinear analysis of reinforced concrete structures. The major advantage of this approach is the simplified finite element model with reduced degrees of freedom and without the need of using displacement functions. The new approach can also trace the formation, development, and location of cracks in the reinforced concrete structures. The results of numerical analyses of structural examples showed that this new approach gave satisfactory results to the engineering problems in comparison with those obtained using the more complicated FE package ANSYS8.0.
Dynamics of the Immune Reaction to Pancreatic Cancer from Inception to Invasion
Authors: Clark Carolyn E. , Hingorani Sunil R. , Mick Rosemarie , Combs Chelsea , Tuveson David A. , Vonderheide Robert H.
Published: October 2007

The dynamics of cancer immunosurveillance remain incompletely understood, hampering efforts to develop immunotherapy of cancer. We evaluated the evolving in vivo immune response to a spontaneous tumor in a genetically defined mouse model of pancreatic ductal adenocarcinoma from the inception of preinvasive disease to invasive cancer. We observed a prominent leukocytic infiltration even around the lowest grade preinvasive lesions, but immunosuppressive cells, including tumor-associated macrophages, myeloid-derived suppressor cells (MDSC), and regulatory T cells (Treg), dominated the early response and persisted through invasive cancer. Effector T cells, however, were scarce in preinvasive lesions, found in only a subset of advanced cancers, and showed no evidence of activation. The lack of tumor-infiltrating effector T cells strongly correlated with the presence of intratumoral MDSC with a near mutual exclusion. In vitro, we found that MDSC suppressed T-cell proliferation. Overall, our results show that suppressive cells of the host immune system appear early during pancreatic tumorigenesis, preceding and outweighing antitumor cellular immunity, and likely contribute to disease progression. Thus, in contrast to the hypothesis that an early “elimination phase” of cancer immunosurveillance is eventually overwhelmed by a growing invasive tumor, our findings suggest that productive tumor immunity may be undermined from the start. Efforts to test potent inhibitors of MDSC, tumor-associated macrophages, and Treg, particularly early in the disease represent important next steps for developing novel immunotherapy of cancer. [Cancer Res 2007;67(19):9518–27]
Cancer Stem Cells in Radiation Resistance
Authors: Rich Jeremy N.
Published: October 2007

Highly tumorigenic subpopulations of several solid cancers share characteristics with somatic stem cells. We showed recently that cancer stem cells, or tumor-initiating cells, derived from human glioblastoma surgical specimens and xenografts display resistance to radiation due to increased activation of the DNA damage checkpoint. We additionally showed that these same tumor subpopulations promote tumor angiogenesis through increased expression of vascular endothelial growth factor. These studies and subsequent reports from other researchers support critical roles for cancer stem cells in determining tumor response to therapy. Hypoxia and stem cell maintenance pathways may provide therapeutic targets to sensitize cancer stem cells to cytotoxic therapies to improve cancer patient treatments. [Cancer Res 2007;67(19):8980–4]
Triptolide Induces Pancreatic Cancer Cell Death via Inhibition of Heat Shock Protein 70
Authors: Phillips Phoebe A. , Dudeja Vikas , McCarroll Joshua A. , Borja-Cacho Daniel , Dawra Rajinder K. , Grizzle William E. , Vickers Selwyn M. , Saluja Ashok K.
Published: October 2007

Pancreatic cancer is highly resistant to current chemotherapy agents. We therefore examined the effects of triptolide (a diterpenoid triepoxide) on pancreatic cancer growth and local-regional tumor spread using an orthotopic model of pancreatic cancer. We have recently shown that an increased level of HSP70 in pancreatic cancer cells confers resistance to apoptosis and that inhibiting HSP70 induces apoptosis in these cells. In addition, triptolide was recently identified as part of a small molecule screen, as a regulator of the human heat shock response. Therefore, our aims were to examine the effects of triptolide on (a) pancreatic cancer cells by assessing viability and apoptosis, (b) pancreatic cancer growth and local invasion in vivo, and (c) HSP70 levels in pancreatic cancer cells. Incubation of PANC-1 and MiaPaCa-2 cells with triptolide (50–200 nmol/L) significantly reduced cell viability, but had no effect on the viability of normal pancreatic ductal cells. Triptolide induced apoptosis (assessed by Annexin V, caspase-3, and terminal nucleotidyl transferase–mediated nick end labeling) and decreased HSP70 mRNA and protein levels in both cell lines. Triptolide (0.2 mg/kg/d for 60 days) administered in vivo decreased pancreatic cancer growth and significantly decreased local-regional tumor spread. The control group of mice had extensive local invasion into adjacent organs, including the spleen, liver, kidney, and small intestine. Triptolide causes pancreatic cancer cell death in vitro and in vivo by induction of apoptosis and its mechanism of action is mediated via the inhibition of HSP70. Triptolide is a potential therapeutic agent that can be used to prevent the progression and metastases of pancreatic cancer. [Cancer Res 2007;67(19):9407–16]
Genetic Heterogeneity among Fanconi Anemia Heterozygotes and Risk of Cancer
Authors: Berwick Marianne , Satagopan Jaya M. , Ben-Porat Leah , Carlson Ann , Mah Katherine , Henry Rashida , Diotti Raffaella , Milton Kelly , Pujara Kanan , Landers Tom , Dev Batish Sat , Morales José , Schindler Detlev , Hanenberg Helmut , Hromas Robert , Levran Orna , Auerbach Arleen D.
Published: October 2007

Fanconi anemia (FA) is a rare autosomal recessive disease characterized by a greatly increased risk of cancer among those diagnosed with the syndrome. The question as to whether FA heterozygotes are at increased risk for cancer is of great importance to those at risk for being a carrier. To address this question, we formed a cohort of grandparents of probands identified through the International Fanconi Anemia Registry. We obtained informed consent, a short questionnaire, and either blood or buccal swab DNA. After diagnosis of the proband was confirmed and complementation studies or DNA sequencing on the proband were completed, mutation analyses of the putative carriers and noncarriers was carried out. Standardized incidence ratios (SIR) were calculated to compare the observed cancer incidence of the grandparents and other relatives with the expected rates of cancer, using the Surveillance, Epidemiology, and End Results registries and the Connecticut Cancer registry. In the 944 study subjects who participated (784 grandparents and 160 other relatives), there was no suggestion of an increase in overall cancer incidence. On the other hand, a significantly higher rate of breast cancer than expected was observed among carrier grandmothers [SIR, 1.7; 95% confidence interval (95% CI), 1.1–2.7]. Among the grandmothers, those who were carriers of FANCC mutations were found to be at highest risk (SIR, 2.4; 95% CI, 1.1–5.2). Overall, there was no increased risk for cancer among FA heterozygotes in this study of Fanconi relatives, although there is some evidence that FANCC mutations are possibly breast cancer susceptibility alleles. [Cancer Res 2007;67(19):9591–6]
Association Study of 69 Genes in the Ret Pathway Identifies Low-penetrance Loci in Sporadic Medullary Thyroid Carcinoma
Authors: Ruiz-Llorente Sergio , Montero-Conde Cristina , Milne Roger L. , Moya Christian M. , Cebrián Arancha , Letón Rocío , Cascón Alberto , Mercadillo Fátima , Landa Iñigo , Borrego Salud , de Nanclares Guiomar Pérez , Álvarez-Escolá Cristina , Díaz-Pérez José Ángel , Carracedo Ángel , Urioste Miguel , González-Neira Anna , Benítez Javier , Santisteban Pilar , Dopazo Joaquín , Ponder Bruce A. , Robledo Mercedes ,   the Medullary Thyroid Carcinoma Clinical Group
Published: October 2007

To date, few association studies have been done to better understand the genetic basis for the development of sporadic medullary thyroid carcinoma (sMTC). To identify additional low-penetrance genes, we have done a two-stage case-control study in two European populations using high-throughput genotyping. We selected 417 single nucleotide polymorphisms (SNP) belonging to 69 genes either related to RET signaling pathway/functions or involved in key processes for cancer development. TagSNPs and functional variants were included where possible. These SNPs were initially studied in the largest known series of sMTC cases (n = 266) and controls (n = 422), all of Spanish origin. In stage II, an independent British series of 155 sMTC patients and 531 controls was included to validate the previous results. Associations were assessed by an exhaustive analysis of individual SNPs but also considering gene- and linkage disequilibrium–based haplotypes. This strategy allowed us to identify seven low-penetrance genes, six of them (STAT1, AURKA, BCL2, CDKN2B, CDK6, and COMT) consistently associated with sMTC risk in the two case-control series and a seventh (HRAS) with individual SNPs and haplotypes associated with sMTC in the Spanish data set. The potential role of CDKN2B was confirmed by a functional assay showing a role of a SNP (rs7044859) in the promoter region in altering the binding of the transcription factor HNF1. These results highlight the utility of association studies using homogeneous series of cases for better understanding complex diseases. [Cancer Res 2007;67(19):9561–7]
Enhancement of Antibody-Dependent Mechanisms of Tumor Cell Lysis by a Targeted Activator of Complement
Authors: Imai Masaki , Ohta Rieko , Varela Juan C. , Song Hongbin , Tomlinson Stephen
Published: October 2007

Complement inhibitors expressed on tumor cells provide a hindrance to the therapeutic efficacy of some monoclonal antibodies (mAb). We investigated a novel strategy to overwhelm complement inhibitor activity and amplify complement activation on tumor cells. The C3-binding domain of human complement receptor 2 (CR2; CD21) was linked to the complement-activating Fc region of human IgG1 (CR2-Fc), and the ability of the construct to target and amplify complement deposition on tumor cells was investigated. CR2 binds C3 activation fragments, and CR2-Fc targeted tumor cells by binding to C3 initially deposited by a tumor-specific antibody. Complement deposition on Du145 cells (human prostate cancer cell line) and anti-MUC1 mAb-mediated complement-dependent lysis of Du145 cells were significantly enhanced by CR2-Fc. Anti-MUC1 antibody-dependent cell-mediated cytotoxicity of Du145 by human peripheral blood mononuclear cells was also significantly enhanced by CR2-Fc in both the presence and the absence of complement. Radiolabeled CR2-Fc targeted to s.c. Du145 tumors in nude mice treated with anti-MUC1 mAb, validating the targeting strategy in vivo. A metastatic model was used to investigate the effect of CR2-Fc in a therapeutic paradigm. Administration of CR2-Fc together with mAb therapy significantly improved long-term survival of nude mice challenged with an i.v. injection of EL4 cells. The data show that CR2-Fc enhances the therapeutic efficacy of antibody therapy, and the construct may provide particular benefits under conditions of limiting antibody concentration or low tumor antigen density. [Cancer Res 2007;67(19):9535–41]
Glucuronidation of Nicotine and Cotinine by UGT2B10: Loss of Function by the UGT2B10 Codon 67 (Asp>Tyr) Polymorphism
Authors: Chen Gang , Blevins-Primeau Andrea S. , Dellinger Ryan W. , Muscat Joshua E. , Lazarus Philip
Published: October 2007

Nicotine, the major addicting agent in tobacco and tobacco smoke, undergoes a complex metabolic pathway, with ∼22% of nicotine urinary metabolites in the form of phase II N-glucuronidated compounds. Recent studies have shown that UGT2B10 is a major enzyme involved in the N-glucuronidation of several tobacco-specific nitrosamines. In the present study, microsomes of UGT2B10-overexpressing HEK293 cells exhibited high N-glucuronidation activity against both nicotine and cotinine with apparent KM's that were 37- and 3-fold lower than that observed for microsomes of UGT1A4-overexpressing cells against nicotine and cotinine, respectively. The KM of microsomes from wild-type (WT) UGT2B10-overexpressing cells for nicotine and cotinine was similar to that observed for human liver microsomes (HLM) against both substrates. The level of glucuronidated nicotine or cotinine in 112 HLM samples was correlated with UGT2B10 genotype; the levels of nicotine- and cotinine-glucuronide were 21% to 30% lower in specimens from subjects with the UGT2B10 (*1/*2) genotype compared with specimens from subjects with the WT UGT2B10 (*1/*1) genotype; a 5- and 16-fold lower level of nicotine- and cotinine-glucuronide formation, respectively, was observed in HLM from subjects with the UGT2B10 (*2/*2) genotype. In contrast to the relatively high activity observed for cells overexpressing WT UGT2B10 in vitro, little or no glucuronidation was observed for microsomes from cells overexpressing the UGT2B10*2 variant against either nicotine or cotinine. These data suggest that UGT2B10 is the major hepatic enzyme involved in nicotine/cotinine glucuronidation and that the UGT2B10*2 variant significantly reduces nicotine- and cotinine-N-glucuronidation formation and plays an important role in nicotine metabolism and elimination. [Cancer Res 2007;67(19):9024–9]
Molecular Analysis of Colorectal Cancer Tumors from Patients with Mismatch Repair–Proficient Hereditary Nonpolyposis Colorectal Cancer Suggests Novel Carcinogenic Pathways
Authors: Sánchez-de-Abajo Ana , de la Hoya Miguel , van Puijenbroek Marjo , Tosar Alicia , López-Asenjo J.A. , Díaz-Rubio Eduardo , Morreau Hans , Caldes Trinidad
Published: October 2007

Purpose: A subset of colorectal cancers (CRC) arises in families that, despite fulfilling clinical criteria for hereditary nonpolyposis colorectal cancer (HNPCC), do not show evidence of a mismatch repair (MMR) deficiency. The main objective of this study was to characterize these tumors at the molecular level.

Experimental Design: After comprehensive germ line mutation scanning, microsatellite analysis, and MMR protein expressions, we selected a well-defined cohort of 57 colorectal tumors with no evidence of MMR defects. In this group of tumors, we analyzed KRAS, BRAF, and APC somatic mutations, as well as methylguanine methyltransferase (MGMT) and β-catenin expression. We correlated these alterations with clinicopathologic data and explored the relationship between KRAS G > A transitions and lack of MGMT expression.

Results: The mutation profile at the RAS/RAF/MAPK pathway mimics sporadic microsatellite-stable CRCs. We found an average age of diagnosis 10 years older in KRAS-mutated patients (P = 0.001). In addition, we show that KRAS G > A transitions are actively selected by tumors, regardless of MGMT status. Similarities with HNPCC high–microsatellite instability tumors are observed when APC data are analyzed. The APC mutation rate was low and small insertions/deletions accounted for 70% of the alterations. In addition, we found a low frequency of β-catenin nuclear staining. Finally, we did not find evidence of tumors arising in individuals from the same family sharing molecular features.

Conclusions: We show evidence that CRC tumors arising in HNPCC families without MMR alterations have distinctive molecular features. Overall, our work shows that systematic analysis of somatic alterations in a well-defined subset of CRCs is a good approach to provide new insights into the mechanisms of colorectal carcinogenesis.
Herpes Simplex Virus Us3(−) Mutant as Oncolytic Strategy and Synergizes with Phosphatidylinositol 3-Kinase-Akt–Targeting Molecular Therapeutics
Authors: Liu Ta-Chiang , Wakimoto Hiroaki , Martuza Robert L. , Rabkin Samuel D.
Published: October 2007

Purpose: Oncolytic herpes simplex virus (HSV) vectors have shown safety in clinical trials, but efficacy remains unsatisfactory. Novel HSV vectors that possess tumor selectivity with enhanced potency are therefore needed. The gene product of HSV Us3 protects virus-infected cells from apoptosis, a cellular pathway frequently dysfunctional in tumors. We hypothesized that Us3 mutants, whose replication would be inhibited by apoptosis in normal cells, would be selective for tumor cells.

Experimental Design: HSV mutants G207 (ribonucleotide reductase−/γ34.5−), R7041 (Us3−), and R7306 (Us3 revertant) were tested in normal and tumor cells for viral replication, antitumoral potency, apoptosis induction, and Akt activation. Safety and biodistribution after systemic administration and antitumoral efficacy after intratumoral (i.t.) or i.v. administration were examined.

Results: Us3 deletion results in up to 3-log replication inhibition in normal cells, which correlates with enhanced apoptosis induction. In contrast, R7041 replicates very well in tumor cells, showing 1 to 2 log greater yield than G207. In vivo, R7041 shows no signs of toxicity after systemic delivery in both immunocompetent and immunodeficient mice and shows preferential and prolonged replication in tumors compared with normal tissues. R7041 displays significant antitumoral efficacy after i.t. or i.v. administration. An additional feature of Us3 mutants is enhanced Akt activation compared with wild-type infection, which sensitizes cells to phosphatidylinositol 3-kinase-Akt inhibitors (LY294002, Akt inhibitor IV), shown by synergistic antitumoral activity in vitro and enhanced efficacy in vivo.

Conclusions: Us3 deletion confers enhanced tumor selectivity and antitumoral potency on herpes simplex virus-1 and provides for a novel mechanism of combination therapy with phosphatidylinositol 3-kinase-Akt–targeting molecular therapeutics.
Eotaxin-2 and Colorectal Cancer: A Potential Target for Immune Therapy
Authors: Cheadle Eleanor J. , Riyad Kallingal , Subar Daren , Rothwell Dominic G. , Ashton Garry , Batha Hayley , Sherlock David J. , Hawkins Robert E. , Gilham David E.
Published: October 2007

Purpose: To study the production of chemokines by colorectal hepatic metastases.

Experimental Design: Biopsies of resected colorectal hepatic metastases and nonneoplastic adjacent liver tissue were screened for chemokines using protein arrays and results were confirmed by ELISA and immunohistochemistry.

Results: Two chemokines, eotaxin-2 and MCP-1, were found at elevated levels within the tumor biopsy compared with adjacent liver. The relative increase in expression from tumor was much higher for eotaxin-2 than MCP-1, with 10 of 25 donors having a >100-fold increase in expression compared with 0 of 24 donors for MCP-1. In a parallel analysis, eotaxin-2 was also found at elevated levels in the tumor region of primary colorectal cancer biopsies. Immunohistochemical staining indicated that carcinoembryonic antigen–positive tumor cells stained strongly for eotaxin-2, implicating these cells as the predominant source of the chemokine. In vitro studies confirmed that several colorectal tumor lines produce eotaxin-2 and that secretion of this chemokine could be depressed by IFN-γ and enhanced by the Th2-type cytokines interleukin-4 and interleukin-13. Jurkat T cells were engineered to express the receptor for eotaxin-2 (CCR3). These cells effectively migrated in response to eotaxin-2 protein, suggesting that immune cells gene modified to express a chemokine receptor may have improved abilities to home to tumor.

Conclusions: Taken together, these observations confirm eotaxin-2 as a chemokine strongly associated with primary and metastatic tumors of colorectal origin. Furthermore, the importance of this result may be a useful tool in the development of targeted therapeutic approaches to colorectal tumors.
High-resolution oligonucleotide array-CGH pinpoints genes involved in cryptic losses in chronic lymphocytic leukemia
Authors: Tyybäkinoja A. , Vilpo J. , Knuutila S.
Published: October 2007

Although recurrent chromosomal alterations occur in chronic lymphocytic leukemia (CLL), relatively few affected tumor suppressors and oncogenes have been implicated. To improve genetic characterization of CLL, we performed high-resolution gene copy number analysis of 20 CLL patients using oligonucleotide array comparative genomic hybridization (aCGH). The most recurrent losses were observed in 13q and 11q with variable sizes. The 11q losses varied between 7.44 Mb and 41.72 Mb in size and targeted ATM among others. Lost regions in 13q were generally smaller, spanning from 0.79 Mb to 29.33 Mb. The minimal common region (158 kb) in 13q14.3, which was also homozygously deleted in some cases, harbored five genes: TRIM13, KCNRG, DLEU2, DLEU1, and FAM10A4. Additionally, two micro-RNA genes (MIRN15A and MIRN16-1) locate to the region. New cryptic losses were detected in 1q23.2→q23.3, 3p21.31, 16pter→p13.3, 17p13.3→p13.2, 17q25.3→qter, and 22q11.22. In conclusion, our oligonucleotide aCGH study revealed novel aberrations and provided detailed genomic profiles of the altered regions.
Tiling resolution array CGH of dic(7;9)(p11∼13;p11∼13) in B-cell precursor acute lymphoblastic leukemia reveals clustered breakpoints at 7p11.2∼12.1 and 9p13.1
Authors: Lundin C. , Heidenblad M. , Strömbeck B. , Borg Å. , Hovland R. , Heim S. , Johansson B.
Published: October 2007

The dic(7;9)(p11∼13;p11∼13) is a recurrent chromosomal abnormality in acute lymphoblastic leukemia (ALL), mainly of B-lineage. Although more than 20 dic(7;9)-positive ALLs have been reported to date, the molecular genetic consequences of this aberration are unknown. We performed tiling resolution (32K) genome-wide array-based comparative genomic hybridization (array CGH) analysis of three cases with dic(7;9) in order to characterize the breakpoints on 7p and 9p. The analysis showed a clustering of breakpoints within 9p13.1 in all three cases and within 7p11.2 in two cases; the array CGH revealed two different breakpoints – 7p12.1 and 7p14.1 – in the remaining case. Based on these findings the abnormality should hence be designated dic(7;9)(p11.2∼12.1;p13.1). Locus-specific fluorescence in situhybridization analysis of one of the cases narrowed down the 7p11.2 breakpoint to a <500-kb segment in this sub-band, a region containing three known genes. Unfortunately, lack of material precluded further molecular genetic studies, and it thus remains unknown whether the pathogenetically important outcome of the dic(7;9) is formation of a chimeric gene or loss of 7p and/or 9p material.
Neocentric small supernumerary marker chromosomes (sSMC) – three more cases and review of the literature
Authors: Liehr T. , Utine G.E. , Trautmann U. , Rauch A. , Kuechler A. , Pietracz J. , Bocian E. , Kosyakova N. , Mrasek K. , Boduroglu K. , Weise A. , Aktas D.
Published: October 2007

Here we report on three new patients with neocentric small supernumerary marker chromosomes (sSMC) derived from chromosome 2, 13 and 15, respectively. The sSMC(13) and sSMC(15) had inverted duplicated shapes and the sSMC(2) a ring chromosome shape. All three cases were clinically severely abnormal. A review of the available sSMC literature revealed that up to the present 73 neocentric sSMC cases including these three new cases have been reported. Seven of these cases were not characterized morphologically; in the remainder, 80% had an inverted duplication, 17% a ring and 3% a minute shape. 81% of the reported neocentric sSMC carriers showed severe, 12% moderate and 8% no clinical abnormalities. In summary, we report three more neocentric sSMC cases, provide a review on all up to now published cases, highlight their special characteristics and compare them to centric sSMC.
Human chromosome 1 satellite 3 DNA is decondensed, demethylated and transcribed in senescent cells and in A431 epithelial carcinoma cells
Authors: Enukashvily N.I. , Donev R. , Waisertreiger I.S.-R. , Podgornaya O.I.
Published: October 2007

Constitutive heterochromatin mainly consists of different classes of satellite DNAs and is defined as a transcriptionally inactive chromatin that remains compact throughout the cell cycle. The aim of this work was to investigate the level of condensation, methylation and transcriptional status of centromeric (alphoid DNA) and pericentromeric satellites (human satellite 3, HS3) in tissues (lymphocytes, placenta cells) and in cultured primary (MRC5, VH-10, AT2Sp) and malignant (A431) cells. We found that alphoid DNA remained condensed and heavily methylated in all the cell types. The HS3 of chromosome 1 (HS3-1) but not of chromosome 9 (HS3-9) was strongly decondensed and demethylated in A431 cells. The same observation was made for aged embryonic lung (MRC5) and juvenile foreskin (VH-10) fibroblasts obtained at late passages (32nd and 23rd, respectively). Decondensation was also found in ataxia telangiectasia AT2Sp fibroblasts at the 16th passage. One of the manifestations of the disease is premature aging. The level of HS3-1 decondensation was higher in aged primary fibroblasts as compared to A431. The HS3-1 extended into the territory of neighbouring chromosomes. An RT-PCR product was detected in A431 and senescent MRC5 fibroblasts using primers specific for HS3-1. The RNA was polyadenylated and transcribed from the reverse chain. Our results demonstrate the involvement of satellite DNA in associations between human chromosomes and intermingling of chromosome territories. The invading satellite DNA can undergo decondensation to a certain level. This process is accompanied by demethylation and transcription.
Cutaneous Manifestations in Inflammatory Bowel Diseases: Eight Cases of Psoriasis Induced by Anti-Tumor-Necrosis-Factor Antibody Therapy
Authors: Passarini Beatrice , Infusino Salvatore Domenico , Barbieri Elena , Varotti Elisa , Gionchetti Paolo , Rizzello Fernando , Morselli Claudia , Tambasco Rosy , Campieri Massimo
Published: October 2007

Background: Ulcerous rectocolitis and Crohn’s disease are the best known forms of inflammatory bowel disease (IBD). Skin manifestations are not uncommon in IBD and may be divided into specific cutaneous signs, aspecific cutaneous signs, and cutaneous signs caused by drugs used for IBD therapy. The specific signs (fistulas, rhagades and ulcers) are the result of the diffusion of the intestinal inflammatory process into the skin. Aspecific cutaneous signs (stomatic aphthosis, erythema nodosum, pyoderma gangrenosum, Sweet’s syndrome, vasculitis, bullous diseases) are quite frequently found in those suffering from IBD, but also in apparently healthy subjects, and may sometimes be the first sign of the intestinal disease. Cutaneous manifestations due to drugs vary in clinical aspect and are the direct consequence of the therapies adopted, which in IBD patients can be quite numerous: steroids, immunosuppressants, 5-aminosalicylic acid, biological agents, antibiotics. Objective and Methods:Due to the frequent finding of cutaneous manifestations in patients affected by IBD, a collaboration was set up between the Dermatological Clinic of the University of Bologna and the Center for the Study of IBD of the same university hospital. The aim was to diagnose the cutaneous signs appearing during IBD and to establish their etiopathogenesis in order to assess whether they were the result of epiphenomena of the IBD or side effects of the therapies adopted. Results: The cutaneous manifestations we observed can be divided into three distinct groups: signs that were specific to the basic disease, aspecific signs and finally signs attributable to the drugs used for therapy. Particular attention was given to the aspecific signs and those consequential to therapy. The aspecific cutaneous signs seen in our clinic generally reflect those reported in the literature. The cutaneous manifestations due to drugs were further divided into three groups: rosacea, acneiform dermatitis and psoriasis-like dermatitis. The most notable aspect of our series is the high number of patients presenting psoriasiform-type dermatitides with a generally widespread diffusion. Conclusion: We would like to draw attention to the fact that all patients with psoriasis had been undergoing treatment with drugs inhibiting tumor necrosis factor α (TNF-α) as part of IBD therapy. In all cases, the cutaneous reaction started after the third or fourth infusion of the biological drug. Anti-TNF-α agents have also been successfully used to treat psoriasis in the last few years. The reason for this apparently paradoxical effect of the therapy is still unclear.
Low-Dose X-Ray Depilatory Treatment Induces Trichoblastic Tumors of the Scalp
Authors: Fazaa B. , Cribier B. , Zaraa I. , Zermani R. , Zeglaoui F. , Zouari B. , Ben Jilani S. , Maalej M. , Kamoun M.R.
Published: October 2007

Background: Depilatory radiotherapy for ringworm was largely used before antifungals were available. Patients who underwent this treatment are at high risk of developing scalp tumors or other cancers. The aim of this study was to characterize scalp tumors occurring after X-ray therapy for ringworm. Methods: We included cases of postradiotherapy scalp tumors recorded at the Dermatology Department of the Charles Nicolle Hospital, Tunis between 1988 and 2001. We recorded clinical descriptions and all cases were resubmitted to microscopic analysis. Results: Sixty-one tumors occurred in 33 men and 12 women with a mean age of 49.8 years. Radiodermatitis was present in 21% of patients. Tumors were basal cell carcinomas in 47 cases, trichoblastomas in 10 cases and trichoblastic carcinomas in 4 cases. Twelve patients had 2–5 tumors, with combinations of tumor types in 3 of them. Mean delay of onset of tumors after radiotherapy was 39.4 years in basal cell carcinoma cases, 38.3 years in trichoblastoma cases and 35.6 years in trichoblastic carcinoma cases. Conclusions: This series shows that although basal cell carcinoma is the most frequent tumor in this situation, trichoblastomas are common. We describe, for the first time, radio-induced trichoblastic carcinomas. Trichoblastic tumors have not yet been described in this context because this concept is relatively recent.
The Practical Reality of Using a Patient-Reported Outcome Measure in a Routine Dermatology Clinic
Authors: Salek Sam , Roberts Alaw , Finlay Andrew Y.
Published: October 2007

Objective: To explore whether clinicians used formal quality of life (QoL) information if provided by patients completing the Dermatology Life Quality Index (DLQI) questionnaire and whether the information influenced treatment decision-taking. Methods: The DLQI was completed by adult patients attending a dermatology secondary-care clinic as they arrived at their appointment and given to the clinician. Clinicians recorded whether the DLQI information was used and whether it influenced treatment decision-making. Results: A total of 417 patients attended and 268 questionnaires (64.3%) were completed. The mean DLQI score was 7 (median = 5, range = 0–30). In 64 consultations (28.8%), clinicians used the DLQI information; in 37 of these consultations (57.8%) the DLQI information influenced the clinicians’ treatment decision-making. The mean DLQI score for these 37 consultations was 11.3 (median = 10.0, range = 0–29), indicating a large effect on patients’ QoL. QoL discussion occurred in 85 of 98 consultations observed. The domains of QoL most frequently discussed concerned symptoms (74 consultations) and problems caused by the treatment (24 consultations). Conclusions: Clinicians may use formal QoL information if available in routine clinics. Treatment decisions may be influenced for those with high QoL impairment. Routine systematic assessment of QoL may therefore be of benefit.
Resistance of the Internal Mammary Artery to Restenosis: A Histomorphologic Study of Various Porcine Arteries
Authors: Horstick Georg , Bierbach Benjamin , Schlindwein Peter , Abegunewardene Nico , Vosseler Markus , Bittinger Fernando , Becker Dietmar , Lauterbach Michael , Lehr Hans-Anton , Kempski Oliver
Published: October 2007

Background/Aims: Restenosis after percutaneous transluminal angioplasty (PTA) of the internal mammary artery (IMA) grafts is much less pronounced than in other arteries and venous grafts. The aim of the study was to test whether various arteries respond differently to dilatation. Methods: PTA of the IMA, carotid, renal and circumflex coronary (RCx) arteries was performed in 9 pigs (balloon to artery ratio of 1:1.5). After 8 weeks, angiography was repeated and vessels prepared for histological analysis. Immunohistochemical staining was done to examine proliferative activity (Ki67) and to identify the vasa vasorum of the adventitia (F VIII-RA). Results: The intima-media ratio after PTA was lowest in the IMA (0.06), followed by the carotid (0.27) and renal arteries (0.49) and the RCx (0.69). Proliferation of the intima was seen at 287° of the vessel circumference in the RCx, at 286° in the renal and at 166° in the carotid artery. No proliferative activity was seen in the IMA. The intima-adventitia ratio was lower in the IMA than in the RCx and renal arteries (p < 0.05). Conclusion: Intima proliferation after PTA varies between the different vessels, with best results seen in the IMA. There are differences in remodeling after PTA between muscular, muscular/elastic and elastic arteries.
Phenotypic and Functional Differences between Human Liver Cancer Endothelial Cells and Liver Sinusoidal Endothelial Cells
Authors: Wu Lian Qiu , Zhang Wen Jian , Niu Ji Xiao , Ye Li Ya , Yang Zhi Hua , Grau Georges E. , Lou Jin Ning
Published: October 2007

Background/Aims: The phenotypic and functional characteristics of microvascular endothelial cells derived from human liver cancer (HLCEC) were analyzed in vitro and compared with those of human liver sinusoidal endothelial cells (LSEC). Methods and Results: Flow-cytometric and real-time PCR analysis indicated that expressions of tumor necrosis factor receptor (TNFR) p75, αvβ3 and αvβ5 were increased, while those of TNFR p55 and intercellular-adhesion molecule 1 (ICAM-1) were decreased in HLCEC compared with LSEC. The functional analysis indicated that HLCEC exhibited higher angiogenic ability than LSEC, including proliferation capacity, ability to form capillary-like networks and release of matrix metalloproteinases. In response to tumor necrosis factor, LSEC exhibited a significant dose-dependent cytotoxicity, while HLCEC did not. Moreover, the coagulant and fibrinolytic capacity was increased in HLCEC. In addition, tumor cell adherence was significantly higher on HLCEC than on LSEC, while leukocyte adherence was lower on HLCEC than on LSEC. The cytoadherence of HLCEC was inhibited by antibodies against αvβ3 and αvβ5,and of LSEC by antibodies against ICAM-1. Conclusion: These results indicate that tumor-derived endothelial cells are phenotypically and functionally different from those derived from normal liver tissue. These differences are valuable for understanding tumor angiogenesis and metastasis.
Effect of Prolonged c-Kit Receptor Inhibition by Imatinib Mesylate on the Uterine Contractility of Pregnant Rabbits
Authors: Hutchings G. , Gevaert T. , Deprest J. , Nilius B. , De Ridder D.
Published: October 2007

Background: The c-kit receptor expressed by interstitial cells in the gastrointestinal tract is crucial to their pacemaking function. The function of similar c-kit-expressing myometrial cells is unknown. Methods: Imatinib mesylate, a specific c-kit receptor antagonist, was administered to pregnant New Zealand white rabbits (term = 31 days, n = 35) from day 27 gestation by intramuscular injection twice daily at high (50 µg/kg) or medium (10 µg/kg) dose and compared with a control group injected with vehicle only. In a second phase, two further groups received imatinib at medium or low (1 µg/kg) dose for a longer duration starting from day 18 until delivery. Three does from the latter groups as well as controls underwent myometrial biopsy under general anesthesia after spontaneous vaginal birth. Contractility was recorded by isometric tensiometry. The outcome measures were delay of parturition and in vitro contractility characteristics. Results: High-dose imatinib induced early delivery when compared with the control group (28.6 vs. 30.7 days, p < 0.001). The other groups delivered at term. No effect on in vitro contractility was apparent in any of the groups. Conclusions: c-kit receptor inhibition in pregnant rabbits does not delay significantly the length of gestation or change myometrial contractility in vitro.
Real-Time Quantitative Polymerase Chain Reaction for BCRP, MDR1, and MRP1 mRNA Levels in Lymphocytes and Monocytes
Authors: Moon Young Jin , Zhang Shuzhong , Morris Marilyn E.
Published: October 2007

The expression levels of mRNAs for MDR1 (P-glycoprotein), multidrug resistance-associated proteins (MRP1), and BCRP (breast cancer resistance protein; mitoxantrone resistance protein) were quantitatively determined in lymphocytes and monocytes. Monocytes and lymphocytes were obtained from 3 healthy male and 2 healthy female volunteers. BCRP, MDR1, and MRP1 mRNA levels were determined by real-time quantitative reverse transcriptase polymerase chain reaction. In lymphocytes, relative concentrations of mRNAs for target proteins (BCRP, MDR1, and MRP1) were 1.00 ± 0.478, 9.67 ± 5.53, and 0.116 ± 0.0613 respectively, and in monocytes 1.00 ± 0.854, 0.821 ± 0.263, and 0.090 ± 0.052, respectively. The MDR1 mRNA level was cell type dependent, whereas there was no difference in BCRP and MRP1 expression levels between lymphocytes and monocytes. Comparison of mRNA levels for the three major multidrug-resistant efflux pumps reveals that MDR1 is the predominant form in lymphocytes and BCRP is the predominant form in monocytes.
Involvement of NADPH Oxidases in Cardiac Remodelling and Heart Failure
Authors: Sirker Alexander , Zhang Min , Murdoch Colin , Shah Ajay M.
Published: October 2007

Cardiac remodelling occurs in response to stress, such as chronic hypertension or myocardial infarction, and forms the substrate for subsequent development of heart failure. Key pathophysiological features include ventricular hypertrophy, interstitial fibrosis, contractile dysfunction, and chamber dilatation. Although the molecular mechanisms are complex and not fully defined, substantial evidence now implicates increased oxidative stress as being important. The NADPH oxidase (‘Nox’) enzymes are a particularly important source of reactive oxygen species that are implicated in redox signalling. This article reviews the evidence for an involvement of NADPH oxidases in different aspects of adverse cardiac remodelling. A better understanding of the roles of this complex enzyme family may define novel therapeutic targets for the prevention of heart failure.
A Secretion Trap Screen in Yeast Identifies Protease Inhibitor 16 as a Novel Antihypertrophic Protein Secreted From the Heart
Authors: Frost Robert J.A. , Engelhardt Stefan
Published: October 2007

Background—

Cardiomyocyte hypertrophy is of central importance in the development of congestive heart failure. Whether proteins secreted from the myocardium itself contribute to myocardial hypertrophy is largely unknown.

Methods and Results—

We performed a genetic yeast secretion trap screen using a murine cardiac cDNA library and identified 54 cardiac proteins that contained a secretion signal. When determining their mRNA expression in the myocardium of failing hearts, we found protease inhibitor 16 (PI16) to be strongly upregulated in hypertrophic and failing myocardium. PI16, a 489–amino acid protein with an unknown function, also displayed enhanced expression on the protein level after serum stimulation of primary cardiomyocytes and in failing myocardium. We found PI16 to be secreted rapidly by primary cardiomyocytes into the culture medium, where it inhibited cardiomyocyte growth. RNA interference–mediated suppression of endogenous PI16 in primary cardiomyocytes significantly enhanced cardiomyocyte size. Transgenic mice overexpressing PI16 in a cardiomyocyte-specific manner showed normal cardiac function but had smaller hearts with hypotrophic cardiomyocytes.

Conclusions—

Taken together, we identified 54 putatively secreted cardiac proteins. PI16, a novel protein secreted from the heart, is strongly upregulated early in heart failure and inhibits growth of cardiomyocytes both in vitro and in vivo. PI16 might represent a novel therapeutic target in heart failure.