bluuebunny/crossref_metadata_2025 huggingface dataset:
Swf1p, a Member of the DHHC-CRD Family of Palmitoyltransferases, Regulates the Actin Cytoskeleton and Polarized Secretion Independently of Its DHHC Motif
Authors: Dighe Shubha A. , Kozminski Keith G.
Published: August 2008

Rho and Rab family GTPases play a key role in cytoskeletal organization and vesicular trafficking, but the exact mechanisms by which these GTPases regulate polarized cell growth are incompletely understood. A previous screen for genes that interact with CDC42, which encodes a Rho GTPase, found SWF1/PSL10. Here, we show Swf1p, a member of the DHHC-CRD family of palmitoyltransferases, localizes to actin cables and cortical actin patches in Saccharomyces cerevisiae. Deletion of SWF1 results in misorganization of the actin cytoskeleton and decreased stability of actin filaments in vivo. Cdc42p localization depends upon Swf1p primarily after bud emergence. Importantly, we revealed that the actin regulating activity of Swf1p is independent of its DHHC motif. A swf1 mutant, in which alanine substituted for the cysteine required for the palmitoylation activity of DHHC-CRD proteins, displayed wild-type actin organization and Cdc42p localization. Bgl2p-marked exocytosis was found wild type in this mutant, although invertase secretion was impaired. These data indicate Swf1p has at least two distinct functions, one of which regulates actin organization and Bgl2p-marked secretion. This report is the first to link the function of a DHHC-CRD protein to Cdc42p and the regulation of the actin cytoskeleton.
Structural and Functional Dissection of Mif2p, a Conserved DNA-binding Kinetochore Protein
Authors: Cohen R. L. , Espelin C. W. , De Wulf P. , Sorger P. K. , Harrison S. C. , Simons K. T.
Published: August 2008

Mif2p is the budding-yeast orthologue of the mammalian centromere-binding protein CENP-C. We have mapped domains of Saccharomyces cerevisiae Mif2p and studied the phenotyptic consequences of their deletion. Using chromatin immunoprecipitation (ChIP) and electrophoretic mobility shift assays, we have further shown that Mif2p binds in the CDEIII region of the budding-yeast centromere, probably in close spatial association with Ndc10p. Moreover, ChIP experiments show that Mif2p recruits to yeast kinetochores a substantial subset of inner and outer kinetochore proteins, but not the Ndc80 or Spc105 complexes. We have determined the crystal structure of the C-terminal, dimerization domain of Mif2p. It has a “cupin” fold, extremely similar both in polypeptide chain conformation and in dimer geometry to the dimerization domain of a bacterial transcription factor. The Mif2p dimer seems to be part of an enhanceosome-like structure that nucleates kinetochore assembly in budding yeast.
A Protein Methylation Pathway inChlamydomonasFlagella Is Active during Flagellar Resorption
Authors: Schneider Mark J. , Ulland Megan , Sloboda Roger D.
Published: August 2008

During intraflagellar transport (IFT), the regulation of motor proteins, the loading and unloading of cargo and the turnover of flagellar proteins all occur at the flagellar tip. To begin an analysis of the protein composition of the flagellar tip, we used difference gel electrophoresis to compare long versus short (i.e., regenerating) flagella. The concentration of tip proteins should be higher relative to that of tubulin (which is constant per unit length of the flagellum) in short compared with long flagella. One protein we have identified is the cobalamin-independent form of methionine synthase (MetE). Antibodies to MetE label flagella in a punctate pattern reminiscent of IFT particle staining, and immunoblot analysis reveals that the amount of MetE in flagella is low in full-length flagella, increased in regenerating flagella, and highest in resorbing flagella. Four methylated proteins have been identified in resorbing flagella, using antibodies specific for asymmetrically dimethylated arginine residues. These proteins are found almost exclusively in the axonemal fraction, and the methylated forms of these proteins are essentially absent in full-length and regenerating flagella. Because most cells resorb cilia/flagella before cell division, these data indicate a link between flagellar protein methylation and progression through the cell cycle.
PRODECOMPv3: decompositions of NMR projections for protein backbone and side-chain assignments and structural studies
Authors: Staykova Doroteya K. , Fredriksson Jonas , Billeter Martin
Published: August 2008

Summary: PRODECOMP (projection decomposition) is an implementation of a multi-way decomposition algorithm for the analysis of two-dimensional projections of high-dimensional nuclear magnetic resonance spectra. The newest version, PRODECOMPv3, features a dramatic speedup, more reliable decompositions, a substantial reduction in memory demands, a new graphical user interface and integration into third-party software. These improvements extend the applicability of decompositions to novel types of NMR data on proteins, yielding backbone and side-chain assignments as well as structural information, and therewith enabling complete characterizations of proteins.

Availability: Program, short manual and an example calculation are freely available at www2.chem.gu.se/bcbp/nmr/prodecomp.html

Contact:  doroteya@chem.gu.se;
N-Myc downstream-regulated gene 2 is involved in p53-mediated apoptosis
Authors: Liu Na , Wang Lifeng , Li Xia , Yang Qi , Liu Xinping , Zhang Jing , Zhang Jian , Wu Yousheng , Ji Shaoping , Zhang Yingqi , Yang Angang , Han Hua , Yao Libo
Published: August 2008

The tumor suppressor, p53, is a transcription factor which can modulate the transcription of a number of target genes that are involved in cell-cycle arrest and apoptosis. However, the apoptotic pathway mediated by p53 is not fully understood. Here, we showed that N-Myc downstream-regulated gene 2 (NDRG2) is a new target gene that is regulated by p53. NDRG2 mRNA and protein levels can be upregulated in a p53-dependent manner. The first intron of the NDRG2 gene contains a site that binds p53 directly and mediates wild-type p53-dependent transactivation. In addition, silencing of NDRG2 attenuates p53-mediated apoptosis, whereas over-expression of NDRG2 suppresses tumor cell growth, regardless of the presence or absence of p53. Our results indicate that NDRG2 is a novel p53-inducible target that is involved in the p53-mediated apoptosis pathway.
Detection and Characterization of a Distinct Isolate of Tomato yellow fruit ring virus from Potato
Authors: Golnaraghi A. R. , Pourrahim R. , Ahoonmanesh A. , Zamani-Zadeh H. R. , Farzadfar Sh.
Published: August 2008

A distinct isolate (TY-PF36) of Tomato yellow fruit ring virus (TYFRV) was obtained from potato (Solanum tuberosum) in Iran. Chlorosis and necrotic spots on leaves associated with leaf and stem necrosis symptoms appear on the affected plants. Of 32 plant species and cultivars mechanically inoculated, 24 were susceptible to the virus isolate. The isolate strongly reacted with TYFRV antibodies in enzyme-linked immunosorbent assay (ELISA), but not with the specific antibodies of other tospoviruses tested. TYFRV-specific N gene primers described previously, however, failed to produce DNA fragments from the total RNA extracts of the infected plants in reverse transcription–polymerase chain reaction (RT-PCR). Nucleotide sequencing of the complete N gene and partial L gene of this isolate revealed considerable differences to those reported for TYFRV with identities ranging from 83.9 to 84.2% and 84.9 to 85.4%, respectively. Two specific primers were designed for detecting TY-PF36 using RT-PCR; TY-PF36 was detected in symptomatic field samples of potato, peanut, soybean, and two weed species, Datura metel and D. stramonium.
Auxin-Induced Resistance to Common Scab Disease of Potato Linked to Inhibition of Thaxtomin A Toxicity
Authors: Tegg Robert S. , Gill Warwick M. , Thompson Hannah K. , Davies Noel W. , Ross John J. , Wilson Calum R.
Published: August 2008

Production of the phytotoxin thaxtomin A by pathogenic Streptomyces spp. is essential for induction of common scab disease in potato. Prior studies have shown that foliar application of sublethal concentrations of 2,4-dichlorophenoxyacetic acid (2,4-D) and other auxin or auxin-like compounds significantly reduced severity and occurrence of common scab in subsequently produced tubers. However, the means of disease suppression by these compounds was not known. We confirm the disease suppressive activity of 2,4-D. Detailed tuber physiological examination showed that lenticel numbers, lenticel external dimensions, and periderm thickness and structure, physiological features believed to be critical to Streptomyces scabiei infection, were not substantially changed by 2,4-D treatments, negating a possible mechanism for disease suppression through alteration of these structures. In contrast, our studies show accumulation of 2,4-D in tubers of treated plants occurs and is associated with an enhanced tolerance to thaxtomin A. Applying 2,4-D to cultures of S. scabiei did not significantly alter in vitro growth of the pathogen. Thaxtomin A production by the pathogen was inhibited by 2,4-D, but only at the highest rate tested (1.0 mM), which is at least 200-fold more than is found in 2,4-D treated tubers. These data suggest 2,4-D has no direct effect on the pathogen or its virulence. Confirmatory evidence from studies with Arabidopsis thaliana seedlings demonstrated that the auxins 2,4-D and IAA ameliorate thaxtomin A toxicity. The evidence presented whereby auxin treatment inhibits toxicity of thaxtomin A secreted by the pathogen suggests a novel indirect means of disease suppression.
Natural Infection of Swinglea glutinosa by theCitrus leprosis virus Cytoplasmic Type(CiLV-C) in Colombia
Authors: León M. G. , Becerra C. H. , Freitas-Astúa J. , Salaroli R. B. , Kitajima E. W.
Published: August 2008

Swinglea glutinosa (Blanco) Merr., a perennial plant in the family Rutaceae, is originally from southeast Asia but which is now grown worldwide. In Colombia, it is used as an ornamental and principally as a living fence around rural properties and farms in several regions of the country. Citrus leprosis virus cytoplasmic type (CiLV-C) was recently detected in orange groves of the Colombian Piedmont eastern plains, an area known as the Llanos Orientales (2). Because of the potential for country-wide infection of citrus, some measures are being taken to avoid CiLV-C spread to other regions of Colombia. Further surveys made from June to December 2005 to evaluate the extent of the spread of CiLV-C in the Llanos Orientales revealed some plants in S. glutinosa hedges surrounding citrus orchards exhibiting chlorotic spots and ringspots of varied size on the leaves, similar to those caused by CiLV-C on sweet oranges leaves. These plants were found near citrus orchards in the municipalities of Guamal and in some urban areas of Villavicencio City in the Meta Department. The possibility that these symptoms were caused by CiLV-C was investigated soon after sample collection by the same procedures as described previously for sweet orange (2). In the leaf lesions of S. glutinosa, typical bacilliform particles and dense cytoplasmic viroplasm were found with electron microscopy. Total RNA extracted from symptomatic leaves was subjected to reverse transcription-PCR (RT) using primers (Fwd. 5′GATACGGGACGCATAACA-3′/Rev. 5′-TTCTGGCTCAACATCTGG-3′) that specifically amplify a region within the CiLV-C putative methyltransferase gene and this yielded a single fragment of the expected 402 bp (3). Analysis of the consensus sequence derived from 20 RT-PCR products (GenBank Accession No. EU689106) showed 96% nucleotide and 92% amino acid sequence identity to the sequence of a Brazilian CiLV-C isolate (GenBank Accession Nos. DQ352194.1 and YP_654565.1), respectively. Recently, published work described mite transmission of CiLV-C to some nonrutaceous plants (1), but to our knowledge, this is the first report of a nonCitrus rutaceous plant naturally infected by CiLV-C. Mites found in citrus orchards and previously identified as Brevipalpus phoenicis (Geijskes) (2), which are likely the most important vector of CiLV-C in citrus in Colombia, were observed feeding on healthy and symptomatic S. glutinosa, indicating that S. glutinosa is a host for B. phoenicis. Because the use of S. glutinosa as a living fence or hedge is a common practice in Colombia, CiLV-C-infected S. glutinosa plants may play a role in the epidemiology of leprosis in commercial citrus by serving as an inoculum source for this lethal virus.

References: (1) M. Bastianel et al. Summa Phytopathol. 32:211, 2006. (2) G. A. León et al. Plant Dis. 90:682, 2006. (3) E. C. Locali et al. Plant Dis. 87:1317, 2003.
Genetic Diversity of Spiroplasma citri Strains from Different Regions, Hosts, and Isolation Dates
Authors: Mello A. F. S. , Yokomi R. K. , Melcher U. , Chen J. C. , Wayadande A. C. , Fletcher J.
Published: August 2008

Spiroplasma citri, a phloem-limited pathogen, causes citrus stubborn disease (CSD). Losses due to CSD in California orchards have grown over the past decade. To investigate the possibility of introduction or emergence of a new strain, a study of genetic diversity among S. citri strains from various locations was conducted using random amplified polymorphism DNA-polymerase chain reaction (RAPD-PCR) of 35 strains cultured from 1980 to 1993, and of 35 strains cultured from 2005 to 2006. Analysis using 20 primer pairs revealed considerable diversity among strains. However, no unique genetic signatures were associated with recently collected strains compared with those collected 15 to 28 years ago, and no geographically associated pattern was distinguishable. S. citri strains from carrot and daikon radish contain some unique DNA fragments, suggesting some host plant influence. Multiple strains from single trees also showed genetic diversity. Sequencing of five RAPD bands that differed among strains showed that diversity-related gene sequences include virus fragments, and fragments potentially encoding a membrane lipoprotein, a DNA modification enzyme, and a mobilization element. No differences in colony morphology were observed among the strains. The lack of correlation between PCR patterns and isolation date or collection site is inconsistent with the hypothesis that recent infections are due to the introduction or emergence of novel pathogen strains.
Hairy Nightshade as a Potential Potato leafroll virus (Luteoviridae: Polerovirus) Inoculum Source in Pacific Northwest Potato Ecosystems
Authors: Srinivasan R. , Alvarez J. M.
Published: August 2008

Hairy nightshade, Solanum sarrachoides, is a solanaceous weed found abundantly in Pacific Northwest potato ecosystems. It serves as a reservoir for one of the important potato viruses, Potato leafroll virus (PLRV) (Luteoviridae: Polerovirus), and its most important vector, the green peach aphid, Myzus persicae (Homoptera: Aphididae). Laboratory research indicated an increased green peach aphid settling and performance on S. sarrachoides than on potato. It also revealed that green peach aphids transmitted PLRV more efficiently from S. sarrachoides to potato than from potato to potato. To test the efficiency of S. sarrachoides as an inoculum source in the field, a two season (2004 and 2005) trial was conducted at Kimberly, Idaho. Two inoculum sources, PLRV-infected potato and PLRV-infected S. sarrachoides, were compared in this trial. Green peach aphid density and temporal and spatial PLRV spread were monitored at weekly intervals. Higher densities of green peach aphids were observed on plots with S. sarrachoides and inoculum sources (PLRV-infected S. sarrachoides and potato) than on plots without S. sarrachoides and inoculum sources. PLRV infection in plots with PLRV-infected S. sarrachoides was similar to or slightly higher than in plots with PLRV-infected potato as an inoculum source. Temporal and spatial PLRV spread was similar in plots with either inoculum source. Thus, S. sarrachoides is as efficient as or a better PLRV inoculum source than potato.
Population Structure of Fusarium fujikuroi from California Rice and Water Grass
Authors: Carter L. L. A. , Leslie J. F. , Webster R. K.
Published: August 2008

The recent observance of Fusarium fujikuroi, the causal agent of Bakanae disease of rice, in California provides a unique opportunity to assess the population diversity of an introduced pathogen in a new environment. We collected 172 isolates of this pathogen between 2000 and 2003 from California rice and two from water grass (Echinochloa spp.). Pathogenicity of F. fujikuroi was demonstrated on early water grass (E. oryzoides) and barnyard grass (E. crus-galli) indicating that weed control should be part of Bakanae management programs. Both mating types and six unique amplified fragment length polymorphism haplotypes corresponding to six identified vegetative compatibility groups were detected. The two most frequently isolated haplotypes encompassed 94% of the collected isolates, suggesting that clonal reproduction dominates. Coefficients of similarity between the unique haplotypes ranged from 0.94 to 0.98, and indicate that there is very little genotypic variation in the F. fujikuroi population in California. The near fixation of the MAT-1 idiomorph (observed ratio 170 MAT-1:4 MAT-2), is consistent with a hypothesis of predominant or exclusive asexual reproduction. The low level of introduced genotypic diversity, in conjunction with the asexual reproductive strategy of this population will slow evolutionary processes, including adaptation to the California environment.
Efficacy of Triazole-Based Fungicides for Fusarium Head Blight and Deoxynivalenol Control in Wheat: A Multivariate Meta-Analysis
Authors: Paul P. A. , Lipps P. E. , Hershman D. E. , McMullen M. P. , Draper M. A. , Madden L. V.
Published: August 2008

The effects of propiconazole, prothioconazole, tebuconazole, metconazole, and prothioconazole+tebuconazole (as a tank mix or a formulated premix) on the control of Fusarium head blight index (IND; field or plot-level disease severity) and deoxynivalenol (DON) in wheat were determined. A multivariate random-effects meta-analytical model was fitted to the log-transformed treatment means from over 100 uniform fungicide studies across 11 years and 14 states, and the mean log ratio (relative to the untreated check or tebuconazole mean) was determined as the overall effect size for quantifying fungicide efficacy. Mean log ratios were then transformed to estimate mean percent reduction in IND and DON relative to the untreated check (percent control: [Formula: see text]IND and [Formula: see text]DON) and relative to tebuconazole. All fungicides led to a significant reduction in IND and DON (P < 0.001), although there was substantial between-study variability. Prothioconazole+tebuconazole was the most effective fungicide for IND, with a [Formula: see text]IND of 52%, followed by metconazole (50%), prothioconazole (48%), tebuconazole (40%), and propiconazole (32%). For DON, metconazole was the most effective treatment, with a [Formula: see text]DON of 45%; prothioconazole+tebuconazole and prothioconazole showed similar efficacy, with [Formula: see text]DON values of 42 and 43%, respectively; tebuconazole and propiconazole were the least effective, with [Formula: see text]DON values of 23 and 12%, respectively. All fungicides, with the exception of propiconazole, were significantly more effective than tebuconazole for control of both IND and DON (P < 0.001). Relative to tebuconazole, prothioconazole, metconazole, and tebuconzole+prothioconzole reduced disease index a further 14 to 20% and DON a further 25 to 29%. In general, fungicide efficacy was significantly higher for spring wheat than for soft winter wheat studies; depending on the fungicide, the difference in percent control between spring and soft winter wheat was 5 to 20% for [Formula: see text]IND and 7 to 16% for [Formula: see text]DON. Based on the mean log ratios and between-study variances, the probability that IND or DON in a treated plot from a randomly selected study was lower than that in the check by a fixed margin was determined, which confirmed the superior efficacy of prothioconazole, metconazole, and tebuconzole+prothioconzole for Fusarium head blight disease and toxin control.
Recovery from Cucurbit leaf crumple virus (Family Geminiviridae, Genus Begomovirus) Infection Is an Adaptive Antiviral Response Associated with Changes in Viral Small RNAs
Authors: Hagen C. , Rojas M. R. , Kon T. , Gilbertson R. L.
Published: August 2008

A strong recovery response occurs in cantaloupe (Cucumis melo) and watermelon (Citrullus lanatus) infected with the bipartite begomovirus Cucurbit leaf crumple virus (CuLCrV). This response is characterized by initially severe symptoms, which gradually become attenuated (almost symptomless). An inverse relationship was detected between viral DNA levels and recovery, indicating that recovered tissues had reduced viral titers. Recovered tissues also were resistant to reinfection with CuLCrV; i.e., recovered leaves reinoculated with the virus did not develop symptoms or have an increased level of viral DNA. In contrast, infection of CuLCrV-recovered leaves with the RNA virus, Cucumber mosaic virus (CMV), disrupted recovery, resulting in the development of severe disease symptoms (more severe than those induced by CMV or CuLCrV alone) and increased CuLCrV DNA levels. Small RNAs with homology to CuLCrV DNA were detected in recovered and nonrecovered tissues; as well as in phloem exudates from infected, but not uninfected plants. Levels of these small RNAs were positively correlated with viral titer; thus, recovered tissues had lower levels than symptomatic tissues. In addition, viral DNA from a host that undergoes strong recovery (watermelon) was more highly methylated compared with that from a host that undergoes limited recovery (zucchini). Furthermore, inoculation of CuLCrV-infected zucchini with a construct expressing an inverted repeat of the CuLCrV common region enhanced recovery and reduced viral symptoms and viral DNA levels in newly emerged leaves. Taken together, these results suggest that recovery from CuLCrV infection is an adaptive antiviral defense mechanism, most likely mediated by gene silencing.
Roger Ervin Miller. 23 July 1952 — 6 November 2005
Authors: Baer Tomas , Scoles Giacinto
Published: August 2008

Roger Miller, perhaps the most outstanding experimental physical chemist of his generation, was sadly taken from us at the age of53 years, at the peak of his career.

Roger Ervin Miller was born in Kitchener, Ontario, Canada, on 23 July 1952, the son of Ruth and Donald R. Miller. Roger had one older brother, Gerry, and one younger sister, Patricia. Roger's father worked in a tyre factory and was very innovative and creative in mending items around the house, a trait that he passed on to his children. From a very early age Roger was curious about the world around him. He was also a hard worker, raising rabbits or picking dew worms to earn money so that he could purchase model rockets, the bigger the better. This was the time of Sputnik and the US space programme, Roger being just 5 years old when the first Sputnik satellite was launched in 1957. As Roger explained in a memoir that he began to write in 2000 (but never finished; extracts are reproduced by courtesy of Mrs. Debbie Miller):
Transcription of foreign DNA in Escherichia coli
Authors: Warren René L. , Freeman John D. , Levesque Roger C. , Smailus Duane E. , Flibotte Stephane , Holt Robert A.
Published: August 2008

Propagation of heterologous DNA in E. coli host cells is central to molecular biology. DNA constructs are often engineered for expression of recombinant protein in E. coli, but the extent of incidental transcription arising from natural regulatory sequences in cloned DNA remains underexplored. Here, we have used programmable microarrays and RT-PCR to measure, comprehensively, the transcription of H. influenzae, P. aeruginosa, and human DNA propagating in E. coli as bacterial artificial chromosomes. We find evidence that at least half of all H. influenzae genes are transcribed in E. coli. Highly transcribed genes are principally involved in energy metabolism, and their proximal promoter regions are significantly enriched with E. coli σ70 (also known as RpoD) binding sites. H. influenzae genes acquired from an ancient bacteriophage Mu insertion are also highly transcribed. Compared with H. influenzae, a smaller proportion of P. aeruginosa genes are transcribed in E. coli, and in E. coli there is punctuated transcription of human DNA. The presence of foreign DNA in E. coli disturbs the host transcriptional profile, with expression of the E. coli phage shock protein operon and the flagellar gene cluster being particularly strongly up-regulated. While cross-species transcriptional activation is expected to be enabling for horizontal gene transfer in bacteria, incidental expression of toxic genes can be problematic for DNA cloning. Ongoing characterization of cross-expression will help inform the design of biosynthetic gene clusters and synthetic microbial genomes.
Immunohistochemical detection of CD133 expression in colorectal cancer: A clinicopathological study
Authors: Kojima Motohiro , Ishii Genichiro , Atsumi Naho , Fujii Satoshi , Saito Norio , Ochiai Atsushi
Published: August 2008

CD133 has been reported to be a cancer‐initiating cell marker in colorectal carcinoma. The objective of this study was to evaluate the frequency of CD133 expression in colorectal cancer, the distribution of CD133‐positive cancer cells, and their relationship to clinicopathological features, including survival. An immunohistochemical examination of CD133 expression and a clinicopathological analysis were performed in the 189 consecutive colorectal cancer patients. CD133 expression was seen at the luminal surface of cancer glands mainly with cribriform features. Expression was detected in only 29 of the 189 tumors (15.3%). Of these, 21 tumors (11.1%) showed CD133 overexpression. All 21 tumors with CD133 overexpression were diagnosed as well‐ or moderately‐differentiated adenocarcinoma. There was no difference in the distribution of CD133 expressing cells between the invasive area and surface area. Although there was no difference in recurrence‐free survival between patients with CD133 overexpression and without, the patients with CD133 overexpression had significantly poorer overall survival (P = 0.03). CD133 overexpression is a risk factor for poorer overall survival in patients with well‐ and moderately‐differentiated adenocarcinoma. Expression of this cancer‐initiating cell marker may vary with the histological type of the cancer, and further investigation of the relationship between its expression and clinicopathological features may be necessary. (Cancer Sci 2008; 99: 1578–1583)
Hydrogen peroxide‐mediated nuclear factor κB activation in both liver and tumor cells during initial stages of hepatic metastasis
Authors: Kobayashi Yuki , Nishikawa Makiya , Hyoudou Kenji , Yamashita Fumiyoshi , Hashida Mitsuru
Published: August 2008

Various factors involved in tumor metastasis are regulated by the transcription factor nuclear factor κB (NF‐κB). Because NF‐κB activation may contribute to establishment of hepatic metastasis, its activation in liver cells and tumor cells was separately evaluated in a mouse model of hepatic metastasis. pNF‐κB‐Luc, a firefly luciferase‐expressing plasmid DNA depending on the NF‐κB activity, was injected into the tail vein of mice by the hydrodynamics‐based procedure, a well‐established method for gene transfer to BALB/c male mouse liver. The luciferase activity in the liver was significantly increased by an intraportal inoculation of murine adenocarcinoma colon26 cells, but not of peritoneal macrophages, suggesting that the NF‐κB in liver cells is activated when tumor cells enter the hepatic circulation. Then, colon26 cells stably transfected with pNF‐κB‐Luc were inoculated. The firefly luciferase activity, an indicator of NF‐κB activity in tumor cells, was significantly increased when colon26/NFκB‐Luc cells were inoculated into the portal vein of BALB/c male mice. The NF‐κB activation in both liver and tumor cells was significantly inhibited by injection of catalase derivatives, which have been reported to inhibit hepatic metastasis of tumor cells. These findings indicate for the first time that NF‐κB, a key agent regulating the expression of various molecules involved in tumor metastasis, is activated in both liver and tumor cells during the initial stages of tumor metastasis through a hydrogen peroxide mediated pathway. Thus, the removal of hydrogen peroxide will be a promising approach to treating hepatic metastasis. (Cancer Sci2008; 99: 1546–1552)
Suppression of surfactant protein A by an epidermal growth factor receptor tyrosine kinase inhibitor exacerbates lung inflammation
Authors: Inoue Akira , Xin Hong , Suzuki Takuji , Kanehira Masahiko , Kuroki Yoshio , Fukuhara Tatsuro , Kikuchi Toshiaki , Maemondo Makoto , Nukiwa Toshihiro , Saijo Yasuo
Published: August 2008

Interstitial lung disease (ILD) is reported as a serious adverse event in lung cancer patients treated with gefitinib, an epidermal growth factor receptor tyrosine kinase inhibitor (EGFR‐TKI). However, the mechanisms of ILD associated with gefitinib remain unknown. To address the molecular mechanisms of ILD‐associated gefitinib, we determined the effect of gefitinib treatment on surfactant protein expression in vitro and in vivo. Gefitinib treatment suppressed surfactant protein (SP)‐A expression in H441 human lung adenocarcinoma cells expressing SP‐A, ‐B, ‐C and ‐D by inhibiting epidermal growth factor signal. Next, gefitinib (200 mg/kg) was given p.o. to the mice daily for 1 week. Daily administration of gefitinib gradually reduced SP‐A level in the bronchoalveolar lavage fluid. When lipopolysaccharide (LPS) was instilled intratracheally to the mice pretreated with gefitinib for 1 week, lung inflammation by LPS was exacerbated and prolonged. This exacerbation of lung inflammation was rescued by intranasal administration of SP‐A. These results demonstrated that pretreatment with gefitinib exacerbated LPS‐induced lung inflammation by reducing SP‐A expression in the lung. This study suggests that epidermal growth factor receptor tyrosine kinase inhibitor may reduce SP‐A expression in the lungs of lung cancer patients and thus patients treated with epidermal growth factor receptor tyrosine kinase inhibitor may be susceptible to pathogens. (Cancer Sci 2008; 99: 1679–1684)
Serum levels of soluble major histocompatibility complex (MHC) class I‐related chain A in patients with chronic liver diseases and changes during transcatheter arterial embolization for hepatocellular carcinoma
Authors: Kohga Keisuke , Takehara Tetsuo , Tatsumi Tomohide , Ohkawa Kazuyoshi , Miyagi Takuya , Hiramatsu Naoki , Kanto Tatsuya , Kasugai Tsutomu , Katayama Kazuhiro , Kato Michio , Hayashi Norio
Published: August 2008

Soluble forms of major histocompatibility complex (MHC) class I‐related chain A and B (MICA/B) are increased in the sera of patients with malignancy and impair the antitumor immune response by downregulating expression of their cognate immunoreceptor natural killer group 2, member D (NKG2D). Recently, soluble MICA/B were reported to appear even in some premalignant diseases, raising questions about the impact of soluble MICA/B produced from tumors on the expression of NKG2D. The present study examined soluble MICA/B in chronic liver disease and hepatocellular carcinoma (HCC) and their involvement in the immune‐cell expression of NKG2D during transcatheter arterial embolization for HCC. The levels of soluble MICA/B were significantly higher in chronic liver disease and HCC patients than in healthy volunteers. The progression of liver disease and that of the tumor were independent determinants for soluble MICA/B levels. Immunohistochemistry revealed that MICA/B were expressed not only in HCC tissue but also on hepatocytes in cirrhotic livers. The transcatheter arterial embolization therapy significantly decreased serum levels of soluble MICA, but not soluble MICB, and increased the NKG2D expression on natural killer cells and CD8‐positive T cells; there was an inverse correlation between changes in soluble MICA levels and in NKG2D expression. In conclusion, although soluble MICA/B are produced from both HCC and premalignant cirrhotic livers, therapeutic intervention for HCC can reduce the levels of soluble MICA and thereby upregulate the expression of NKG2D. Cancer therapy may have a beneficial effect on NKG2D‐mediated antitumor immunity. (Cancer Sci 2008; 99: 1643–1649)
Comment on ‘Impact of CYP2D6*10 on recurrence‐free survival in breast cancer patients receiving adjuvant tamoxifen therapy’
Authors: Lash Timothy L. , Ahern Thomas P. , Cronin‐Fenton Deirdre , Garne Jens Peter , Hamilton‐Dutoit Stephen , Sørensen Henrik Toft
Published: August 2008

(Cancer Sci 2008; 99: 1706–1707)
Differential responsiveness of human hepatoma cells versus normal hepatocytes to TRAIL in combination with either histone deacetylase inhibitors or conventional cytostatics
Authors: Dzieran Johanna , Beck James F. , Sonnemann Jürgen
Published: August 2008

Tumor necrosis factor‐related apoptosis‐inducing ligand (TRAIL) is a promising candidate for the treatment of cancer because it elicits cell death in many tumor cells while sparing most normal cells. Liver cancer, however, is largely resistant to TRAIL and, thus, requires sensitization for TRAIL‐mediated cytotoxicity. Sensitization may be achieved by cotreatment with chemotherapeutic agents. In this study, we comparatively investigated the treatment efficacy of TRAIL in combination with histone deacetylase inhibitors (HDI) versus TRAIL in combination with conventional cytostatics in the hepatocellular carcinoma cell line HepG2 and in the childhood hepatoblastoma cell line Huh6. We found that TRAIL resistance could be overcome by cotreatment with the HDI vorinostat, sodium butyrate and MS‐275, but not by cotreatment with the cytostatics carboplatin and etoposide. However, TRAIL combination treatment bears the risk of sensitizing otherwise TRAIL‐resistant normal cells. We thus explored a potential cytotoxic effect of combined HDI/TRAIL treatment in normal hepatocytes: TRAIL in conjunction with HDI did not impose any cytotoxicity on the non‐malignant cells. In searching for the determinants of HDI‐mediated TRAIL sensitization in hepatoma cells, we observed that HDI treatment did not increase cell‐surface expression of proapoptotic TRAIL receptors. Instead, HDI treatment enhanced TRAIL‐induced cleavage of Bid. In conclusion, our data suggest that HDI are potent sensitizers to TRAIL in hepatoma cells and that the combination of HDI and TRAIL is selectively active in hepatoma cells without affecting normal hepatocytes, indicating that the combination of HDI and TRAIL may be an effective approach for the treatment of advanced liver cancer. (Cancer Sci 2008; 99: 1685–1692)
Understanding and exploiting hTERT promoter regulation for diagnosis and treatment of human cancers
Authors: Kyo Satoru , Takakura Masahiro , Fujiwara Toshiyoshi , Inoue Masaki
Published: August 2008

Telomerase activation is a critical step for human carcinogenesis through the maintenance of telomeres, but the activation mechanism during carcinogenesis remains unclear. Transcriptional regulation of the human telomerase reverse transcriptase (hTERT) gene is the major mechanism for cancer‐specific activation of telomerase, and a number of factors have been identified to directly or indirectly regulate the hTERT promoter, including cellular transcriptional activators (c‐Myc, Sp1, HIF‐1, AP2, ER, Ets, etc.) as well as the repressors, most of which comprise tumor suppressor gene products, such as p53, WT1, and Menin. Nevertheless, none of them can clearly account for the cancer specificity of hTERT expression. The chromatin structure via the DNA methylation or modulation of nucleosome histones has recently been suggested to be important for regulation of the hTERT promoter. DNA unmethylation or histone methylation around the transcription start site of the hTERT promoter triggers the recruitment of histone acetyltransferase (HAT) activity, allowing hTERT transcription. These facts prompted us to apply these regulatory mechanisms to cancer diagnostics and therapeutics. Telomerase‐specific replicative adenovirus (Telomelysin, OBP‐301), in which E1A and E1B genes are driven by the hTERT promoter, has been developed as an oncolytic virus that replicates specifically in cancer cells and causes cell death via viral toxicity. Direct administration of Telomelysin was proved to effectively eradicate solid tumors in vivo, without apparent adverse effects. Clinical trials using Telomelysin for cancer patients with progressive stages are currently ongoing. Furthermore, we incorporated green fluorescent protein gene (GFP) into Telomelysin (TelomeScan, OBP‐401). Administration of TelomeScan into the primary tumor enabled the visualization of cancer cells under the cooled charged‐coupled device (CCD) camera, not only in primary tumors but also the metastatic foci. This technology can be applied to intraoperative imaging of metastatic lymphnodes. Thus, we found novel tools for cancer diagnostics and therapeutics by utilizing the hTERT promoter. (Cancer Sci 2008; 99: 1528–1538)
Inhibition of bone and muscle metastases of lung cancer cells by a decrease in the number of monocytes/macrophages
Authors: Hiraoka Koji , Zenmyo Michihisa , Watari Kousuke , Iguchi Haruo , Fotovati Abbas , Kimura Yusuke N. , Hosoi Fumihito , Shoda Takanori , Nagata Kensei , Osada Hiroyuki , Ono Mayumi , Kuwano Michihiko
Published: August 2008

Attention has recently focused on the critical role of inflammatory responses in the tumor stroma that provide favorable conditions for cancer‐cell growth and invasion/metastasis. In particular, macrophages recruited into the tumor stroma and activated, known as tumor‐associated macrophages, are suggested to promote tumorigenesis. In this study, we examined the effect of a decrease in the number of monocytes/macrophages in peripheral blood and the tumor stroma on the development of bone and muscle metastases by lung cancer cells. Treatment with clodronate encapsulated by liposomes (Cl2MDP‐LIP) has been developed for the depletion of monocytes/macrophages in an animal model. Subcutaneous administration of Cl2MDP‐LIP markedly reduced the number of monocytes in peripheral blood, resulting in efficient suppression of both bone metastasis and muscle metastasis when lung cancer HARA‐B cells were injected into the left cardiac ventricle of mice. Treatment with Cl2MDP‐LIP significantly reduced the number of macrophages in tumors and the number of osteoclasts in bone marrow, as well as peripheral monocytes in mice harboring lung cancer cells. In contrast, treatment with an osteoclast‐targeting antibiotic, reveromycin A, inhibited bone metastasis by lung cancer cells, but not muscle metastasis. The survival of human macrophages in culture was found to be specifically blocked by Cl2MDP‐LIP, but not by reveromycin A. Cl2MDP‐LIP thus exerted antimetastatic effects in both bone and muscle whereas reveromycin A did so only in bone. Liposome‐encapsulated bisphosphonate may modulate metastasis through decreasing the number of monocytes/macrophages in both peripheral blood and the tumor stroma, suggesting that tumor‐associated macrophages might be suitable targets for antimetastatic therapy. (Cancer Sci 2008; 99: 1595–1602)
Ammonia release from heated ‘street’ cannabis leaf and its potential toxic effects on cannabis users
Authors: Bloor Roger N. , Wang Tianshu S. , Španěl Patrik , Smith David
Published: August 2008

Aims  To use selected ion flow tube mass spectrometry (SIFT‐MS) to analyse the molecular species emitted by heated ‘street’ cannabis plant material, especially targeting ammonia.

Materials and methods  Samples of ‘street’ cannabis leaf, held under a UK Home Office licence, were prepared by finely chopping and mixing the material. The samples were then heated in commercially available devices. The air containing the released gaseous compounds was sampled into the SIFT‐MS instrument for analysis. Smoke from standard 3% National Institute on Drug Abuse (NIDA) cannabis cigarettes was also analysed.

Findings  For ‘street’ cannabis, ammonia was present in the air samples from the devices at levels approaching 200 parts per million (p.p.m.). This is compared with peak levels of 10 p.p.m. using NIDA samples of known provenance and tetrahydrocannabinol content (3%). Several other compounds were present at lower levels, including acetaldehyde, methanol, acetone, acetic acid and uncharacterized terpenes.

Conclusions  Awareness of the risks of inhaling the smoke directly from burning cannabis has led to the development of a number of alternative methods of delivery, which are claimed to be safer than direct smoking. Ammonia at toxic levels is produced from heating ‘street’ cannabis in these commercially available devices. Thus, the use of these devices to deliver ‘street’ cannabis is now open to question and further research is needed to investigate their safety.
Will we ever find the genes for addiction?
Authors: Buckland Paul R.
Published: August 2008

Aim  To assess the likelihood of finding genes which predispose to addiction and to present this information in a form accessible to the general readership of Addiction.

Methods  Review of the evidence that genetic factors play a significant role in the process of addiction and the proximity of the identification of these factors.

Results  The search for the genetic susceptibility variants for many complex illnesses has been ongoing for decades, with increased pace in the last 20 years. However, until very recently only a small number of such variants have been found. Recent studies have used several thousand samples in genome‐wide association studies and the latest genotyping technology and have reported a growing number of successes, but have highlighted the need for even larger samples and new statistical methods or new experimental approaches to identify fully the genes involved in the disease process. The phenotype for addiction to drugs is not well defined, and the heritability of addiction to drugs of abuse is far from clear and may be small compared to that of many other complex disorders. The absolute requirement for the administration of drugs before addiction can occur, and other environmental factors known to have a major effect, makes the selection of both probands and controls challenging for genetic studies. Many candidate genes put forward so far as susceptibility genes may be unrelated to the underlying process referred to as addiction but, rather, are related to the propensity to take drugs in the first place.

Conclusions  It is the underlying biological process which changes to an alternative state following addiction, which is the target of investigation, and it is not clear that even genome‐wide association studies with sample sizes a magnitude greater than those reported so far would identify the genes involved which have the largest effect. Ultimately, modern neurobiological approaches may identify this process and the genes involved, and even at this stage identifying the susceptibility variants will require both biological as well as genetic analysis.
Marijuana withdrawal and craving: influence of the cannabinoid receptor 1 (CNR1) and fatty acid amide hydrolase (FAAH) genes
Authors: Haughey Heather M. , Marshall Erin , Schacht Joseph P. , Louis Ashleigh , Hutchison Kent E.
Published: August 2008

Aim  To examine whether withdrawal after abstinence and cue‐elicited craving were associated with polymorphisms within two genes involved in regulating the endocannabinoid system, cannabinoid receptor 1 (CNR1) and fatty acid amide hydrolase (FAAH). Two single nucleotide polymorphisms (SNPs) in the CNR1 (rs2023239) and FAAH (rs324420) genes, associated previously with substance abuse and functional changes in cannabinoid regulation, were examined in a sample of daily marijuana smokers.

Participants  Participants were 105 students at the University of Colorado, Boulder between the ages of 18 and 25 years who reported smoking marijuana daily.

Measurements  Participants were assessed once at baseline and again after 5 days of abstinence, during which they were exposed to a cue‐elicited craving paradigm. Outcome measures were withdrawal and craving collected using self‐reported questionnaires. In addition, urine samples were collected at baseline and on day 5 for the purposes of 11‐nor‐9‐carboxy‐Δ9‐tetrahydrocannabinol (THC–COOH) metabolite analysis.

Findings  Between the two sessions, THC–COOH metabolite levels decreased significantly, while measures of withdrawal and craving increased significantly. The CNR1 SNP displayed a significant abstinence × genotype interaction on withdrawal, as well as a main effect on overall levels of craving, while the FAAH SNP displayed a significant abstinence × genotype interaction on craving.

Conclusions  These genetic findings may have both etiological and treatment implications. However, longitudinal studies will be needed to clarify whether these genetic variations influence the trajectory of marijuana use/dependence. The identification of underlying genetic differences in phenotypes such as craving and withdrawal may aid genetically targeted approaches to the treatment of cannabis dependence.
Youth exposure to alcohol use and brand appearances in popular contemporary movies
Authors: Dal Cin Sonya , Worth Keilah A. , Dalton Madeline A. , Sargent James D.
Published: August 2008

Aims  To describe alcohol use and alcohol brand appearances in popular movies and estimate adolescents' exposure to this alcohol‐related content.

Design and setting  Nationally representative, random‐digit dialed survey in the United States and content analysis of alcohol depictions in the top 100 US box office hits each year from 1998 to 2002 and 34 top movies from early 2003.

Participants  A total of 6522 US adolescents aged 10–14 years.

Measurements  Frequency of alcohol use and brand appearances in movies by Motion Picture Association of America (MPAA) rating. Estimated exposure to minutes of movie alcohol use and brand appearances among US adolescents in this age group.

Findings  Most movies (83%, including 56.6% of G/PG‐rated movies) depicted alcohol use and 52% (including 19.2% of G/PG movies) contained at least one alcohol brand appearance, which consisted of branded use by an actor 30.3% of the time. These movies exposed the average US adolescent 10–14 years of age to 5.6 [95% confidence interval (CI) 5.4, 5.7] hours of movie alcohol use and 243.8 (95% CI 238, 250) alcohol brand appearances (5 billion in total), mainly from youth‐rated movies. Exposure to movie alcohol content was significantly higher among African American youth than youth of other races.

Conclusions  Alcohol use and brand appearances are portrayed frequently in popular US movies (which are distributed world‐wide). Children and adolescents in the United States are exposed to hours of alcohol use depictions and numerous brand appearances in movies and most of this exposure is from movies rated for this segment of the population.