bluuebunny/crossref_metadata_2025 huggingface dataset:
Downregulation of connexin 26 in human lung cancer is related to promoter methylation
Authors: Chen Yuan , Hühn Daniela , Knösel Thomas , Pacyna‐Gengelbach Manuela , Deutschmann Nicole , Petersen Iver
Published: August 2004

Cell‐Cell communication via gap junctions plays a key role in carcinogenesis and in growth control. One of the gap junction proteins, Connexin 26 (Cx26) was considered as tumor suppressor in various cancers. In our study, the expression of Cx26 was analyzed in human lung cancer. The reduced mRNA expression was observed in 17 lung cancer cell lines examined by Northern blot analysis and RT‐PCR. In 138 primary carcinomas comprising all subtypes analyzed by immunohistochemistry, 85 cases (62%) exhibited no expression of Cx26, whereas in other 53 cases the Cx26 staining was positive (38%). Additionally, an association between Cx26 protein expression and higher grading of tumors was found in whole tumor samples (p =0.028) but no statistically significant correlations could be observed with tumor stage, tumor size and node status. In squamous cell carcinoma, tumors with higher stage and grading were linked to higher expression of Cx26 (p = 0.015 and 0.017, respectively). To explore the mechanism responsible for the downregulation of Cx26, we treated 2 lung cancer cell lines H2170 and H226 with the demethylation agent 5‐aza‐2′‐deoxycytidine and found the reexpression of Cx26 mRNA. Methylation status of these 2 cell lines was further analyzed by PCR amplification of bisulfite modified DNA and sequencing. A heterogeneous methylation pattern turned out. Our results suggest the inactivation of Cx26 in lung cancer may be explained by promoter methylation.
p53 mutation profiling of multiple esophageal carcinoma using laser capture microdissection to demonstrate field carcinogenesis
Authors: Ito Shuhei , Ohga Takefumi , Saeki Hiroshi , Nakamura Toshihiko , Watanabe Masayuki , Tanaka Shinji , Kakeji Yoshihiro , Maehara Yoshihiko
Published: August 2004

Inactivation of the p53 tumor suppressor gene is one of the most frequent genetic alterations observed in human esophageal carcinomas. In patients with esophageal carcinoma, one of the significant pathological features of the tumor is the presence of multiple lesions within the esophagus. However, the molecular mechanisms involved in the occurrence of multiple lesions have remained elusive. To characterize p53 alterations in multiple esophageal carcinomas and to study their roles in carcinogenesis, we performed p53 immunohistochemical and p53 mutation analyses using laser capture microdissection on surgically resected human esophageal carcinomas from 11 patients: 9 patients with multiple esophageal carcinomas, 1 with an intramural metastasis lesion within the esophagus and 1 with an intraepithelial carcinoma lesion contiguous to the main lesion. In each of the patients with multiple esophageal carcinomas, we examined samples from 1 main lesion and 1 representative concomitant lesion. Molecular analyses of samples from fresh‐frozen normal tissues and tumor tissues of the main lesion (whole tumor) were also performed by the same method. p53 protein accumulation was observed in 16 (72.7%) of 22 lesions from the 11 cases. No p53 mutation was found in normal esophageal tissues. In the 9 cases of multiple esophageal carcinomas, point mutations were detected in the whole tumor in 1 (11.1%) case, in the microdissected tumor samples of main lesions in 3 (33.3%) cases and in the microdissected tumor samples of concomitant lesions in 3 (33.3%) cases. For the microdissected tumor samples, there was a 54.5% (12/22) concordance rate between the results of immunostaining and molecular analysis. In the 8 cases of whole tumors in which a p53 mutation was not observed, 2 cases revealed p53 mutation in the microdissected tumor samples of the main lesion. All 6 cases of multiple esophageal carcinomas that showed a p53 mutation in the microdissected tumor sample had a discordant p53 mutational status between the main and concomitant lesions. In contrast, both the intramural metastasis lesion and the intraepithelial carcinoma contiguous to the main lesion showed p53 mutational patterns identical to those of the main lesions. In conclusion, the analysis of microdissected DNA by laser capture microdissection is useful for characterizing the heterogeneity of the p53 gene mutation in multiple carcinoma lesions that cannot be accurately analyzed in whole esophageal tumor DNA. The finding of different p53 gene mutations among multiple esophageal carcinoma lesions suggest further evidence of multicentric or field carcinogenesis of the human esophagus.
HPV‐associated flat penile lesions in men of a non‐STD hospital population: Less frequent and smaller in size than in male sexual partners of women with CIN
Authors: Bleeker Maaike C.G. , Hogewoning Cornelis J.A. , Voorhorst Feja J. , van den Brule Adriaan J.C. , Berkhof Johannes , Hesselink Albertus T. , Lettink Marjolein , Starink Theo M. , Stoof Tom J. , Snijders Peter J.F. , Meijer Chris J.L.M.
Published: August 2004

Human papillomavirus (HPV) infections and HPV‐associated penile lesions are frequently found in male sexual partners of women with cervical intraepithelial neoplasia (CIN). To determine the significance of these findings, we studied the prevalence of HPV and HPV associated penile lesions in a male hospital population with non‐STD complaints. Penoscopy was performed after application of acetic acid to identify flat lesions, papular lesions, condylomata acuminata and pearly penile papules (PPPs). Presence of HPV DNA in penile scrapes was tested by GP5+6+ PCR. In case of HPV 16 positivity, viral loads were quantified using a LightCycler based real‐time PCR method. Comparing the non‐STD male hospital population (n = 118) with the male sexual partners of women with CIN (n = 238), flat penile lesions were found in 14% vs. 60% and penile HPV in 25% vs. 59% of the men, respectively. We found that the presence of penile HPV and, in case of HPV 16 positivity, higher viral loads were associated with the presence of flat penile lesions. Amongst the HPV‐positive men, flat penile lesions were more common and larger in size in male sexual partners of women with CIN than in the non‐STD hospital population. HPV infections and HPV‐associated flat penile lesions are commonly found in the non‐STD male population. However, these lesions are less frequently present and smaller in size than in male sexual partners of women with CIN. Higher viral loads in penile scrapes of male sexual partners of women with CIN are reflected by a higher prevalence of flat penile lesions and a larger size of these lesions.
TRAF interactions with raft‐like buoyant complexes, better than TRAF rates of degradation, differentiate signaling by CD40 and EBV latent membrane protein 1
Authors: Ardila‐Osorio Hector , Pioche‐Durieu Catherine , Puvion‐Dutilleul Francine , Clausse Bernard , Wiels Joëlle , Miller William , Raab‐Traub Nancy , Busson Pierre
Published: August 2004

The CD40 receptor and the Epstein‐Barr virus oncoprotein LMP1 are both members of the TNF‐receptor family and share several signaling mediators, including TRAF2 and TRAF3. Depending on the cell lineage and stage of maturation, LMP1 and CD40 can have synergistic, antagonist or unrelated effects. Previous publications have suggested that both TRAF2 and TRAF3 move into lipid rafts upon LMP1 expression or CD40 activation, whereas their proteolysis is only enhanced by CD40. However CD40‐induced proteolysis of TRAF2 has only been reported in murine cells, and there are conflicting data regarding translocation of TRAF2 into lipid rafts. We therefore investigated TRAF2 and TRAF3 modifications induced by CD40 and LMP1 signaling in a panel of human cell lines of lymphoid and epithelial origins. Upon CD40 stimulation, a marked redistribution of TRAF2 into the buoyant raft fraction was observed in all cell lines and was often associated with a similar redistribution of TRAF3. In contrast, only TRAF3 was redistributed into the raft fraction upon LMP1 expression. Moreover parallel changes in subcellular distribution of TRAF2 and TRAF3 were recorded by electron microscopy. A significant decrease in TRAF2 and TRAF3 concentrations triggered by CD40 ligation was observed in only 1 cell line and there was no evidence that this decrease was required for the negative feed‐back on JNK activation. TRAF2 redistribution into raft‐like complexes thus appears as the most significant event distinctive of CD40 and LMP1 signaling. On the other hand, the parallel influence of CD40 and LMP1 on TRAF3 redistribution is consistent with functional similarities between the CD40‐TRAF3 and LMP1‐TRAF3 axes.
Occupational risk factors for low grade and high grade glioma: Results from an international case control study of adult brain tumours
Authors: Schlehofer Brigitte , Hettinger Iris , Ryan Philip , Blettner Maria , Preston‐Martin Susan , Little Julian , Arslan Annie , Ahlbom Anders , Giles Graham G. , Howe Geoffrey R. , Ménégoz Francoise , Rodvall Ylva , Choi Won N. , Wahrendorf Jürgen
Published: August 2004

The majority of suspected occupational risk factors for adult brain tumours have yet to be confirmed as etiologically relevant. Within an international case‐control study on brain tumours, lifelong occupational histories and information on exposures to specific substances were obtained by direct interviews to further investigate occupational risk factors for glioma. This is one of the largest studies of brain tumours in adults, including 1,178 cases and 1987 population controls from 8 collaborating study centres matched for age, gender and centre. All occupational information, was aggregated into 16 occupational categories. In a pooled analysis, odds ratios (OR), adjusted for education, were estimated separately for men and women and for high‐grade glioma (HGG) and low‐grade glioma (LGG), focusing especially on 6 categories defined a priori: agricultural, chemical, construction, metal, electrical/electronic and transport. For men, an elevated OR of glioma associated with the category “metal” (OR = 1.24, 95% CI 0.96–1.62) was seen, which appeared to be largely accounted for by LGG (OR = 1.59, 95% CI 1.00–2.52). For the other 5 occupational categories, no elevated risks for glioma were observed. For women the only noteworthy observation for the 6 a priori categories was an inverse association with the “agriculture” category (OR = 0.60, 95% CI 0.36–0.99). Apart from the 6 major categories, women working in food production or food processing (category “food”) showed an increased OR of 1.95 (95% CI 1.04–3.68). None of the 20 substance groups was positively associated with glioma risk. Although some other point estimates were elevated, they lacked statistical significance. The results do not provide evidence of a strong association between occupational exposures and glioma development.
Inhibition of leukemic cell growth by a novel anti‐cancer drug (GUT‐70) from calophyllum brasiliense that acts by induction of apoptosis
Authors: Kimura Shinya , Ito Chihiro , Jyoko Naoto , Segawa Hidekazu , Kuroda Junya , Okada Masayuki , Adachi Souichi , Nakahata Tatsutoshi , Yuasa Takeshi , Filho Valdir Cechinel , Furukawa Hiroshi , Maekawa Taira
Published: August 2004

During our search for cancer chemopreventing compounds derived from plant sources, we discovered that the natural product GUT‐70, isolated from the stem bark of Calophyllum brasiliense collected in Brazil, significantly inhibits the growth of leukemic cells. GUT‐70, characterized as a tricyclic coumarin, 5‐methoxy‐2,2‐dimethyl‐6‐(2‐methyl‐1‐oxo‐2‐butenyl) ‐10‐propyl‐2H,8H‐benzo[1,2‐b;3,4‐b′]dipyran‐8‐one (C23H26O5), inhibited all 6 human leukemic cell lines evaluated, including the P‐glycoprotein overexpressing cell line, in a concentration and time‐dependent manner with IC50 values from 2–5 μM. Furthermore, GUT‐70 did not inhibit colony formation by normal hematopoietic progenitors up to 30 μM and also did not inhibit the proliferation of normal human hepatocytes up to 30 μM. GUT‐70 activated the caspase 2, 3, 8 and 9, and induced the apoptosis in leukemic cells, which was inhibited by caspase inhibitors. GUT‐70 induced anti‐leukemic effects independent of the p53‐p2lWAFl/CIP1 pathway and increased the overall expression of p27KIP1 and p57KIP2, to stop the cell cycle at the G1/S transition. Thus, a novel anti‐cancer drug, GUT‐70 isolated from the stem bark of C. brasiliense induces caspase‐mediated and p53‐independent apoptosis to overcome multidrug resistance and may become a potent leukemia therapeutics.
Alcohol consumption and risk of prostate cancer in middle‐aged men
Authors: Schoonen W. Marieke , Salinas Claudia A. , Kiemeney Lambertus A.L.M. , Stanford Janet L.
Published: August 2004

Alcohol consumption is a modifiable lifestyle factor that may affect prostate cancer risk. Alcohol alters the hormonal milieu and contains chemical substances such as flavonoids (red wine), which may alter tumor cell growth. Data from a population‐based case‐control study in King County, WA, were utilized to evaluate the association of alcohol consumption with prostate cancer in middle‐aged men. A total of 753 newly diagnosed prostate cancer cases, 40–64 years of age, participated in the study. Seven hundred three control subjects, frequency matched to cases by age, were selected through random digit dialing. All participants completed an in‐person interview on lifetime alcohol consumption and other risk factors for prostate cancer. Logistic regression models were used to estimate odds ratios (OR) and assess significance (95% confidence intervals [CI]). All tests of statistical significance were two‐sided. No clear association with prostate cancer risk was seen for overall alcohol consumption. Each additional glass of red wine consumed per week showed a statistically significant 6% decrease in relative risk (OR = 0.94; 95% CI = 0.90–0.98), and there was evidence for a decline in risk estimates across increasing categories of red wine intake (trend p = 0.02). No clear associations were seen for consumption of beer or liquor. Our present study suggests that consumption of beer or liquor is not associated with prostate cancer. There may be, however, a reduced relative risk associated with increasing level of red wine consumption. Further research is needed to evaluate the potential negative association between red wine intake and prostate cancer risk.
Aberrant promoter methylation of human DAB2 interactive protein (hDAB2IP) gene in lung cancers
Authors: Yano Masaaki , Toyooka Shinichi , Tsukuda Kazunori , Dote Hideaki , Ouchida Mamoru , Hanabata Tetsuro , Aoe Motoi , Date Hiroshi , Gazdar Adi F. , Shimizu Nobuyoshi
Published: August 2004

The human DOC‐2/DAB2 interactive protein gene (hDAB2IP) is a novel member of the Ras GTPase‐activating gene family that is known to act as a tumor suppressor gene and is inactivated by methylation in prostate and breast cancers. We established previously a methylation‐specific PCR (MSP) for the promoter region (m2a and m2b regions) of hDAB2IP and examined hDAB2IP methylation status in breast cancers. We analyzed the methylation and expression status of hDAB2IP in lung cancers. The methylation status of hDAB2IP was examined in lung cancer cell lines using bisulfite sequencing and MSP. Expression was examined using conventional and real‐time RT‐PCR, and methylation was found to be inversely correlated with expression, confirming that the MSP can also be used to examine hDAB2IP methylation status in lung cancers. Aberrant methylation was detected at the m2a region in 19 of 47 lung cancer cell lines (40%) and 26 of 70 primary tumors (37%) and at the m2b in 16 lines (34%) and 25 of 70 tumors (36%). Gene expression was restored in methylated cell lines supplemented with 5‐aza‐2′‐deoxycytidine, confirming that methylation was responsible for downregulation. We also examined the relationship between hDAB2IP methylation and clinico‐pathological features of the lung cancers and found that hDAB2IP methylation was associated with advanced disease stage. Our results demonstrate that hDAB2IP methylation is frequently present in lung cancers and plays a key role in hDAB2IP silencing. hDAB2IP methylation could be used as a biomarker for disease stage, reflecting the degree of clinico‐pathological malignancy of lung cancer.
Xenogeneic immunization in mice using HER2 DNA delivered by an adenoviral vector
Authors: Gallo Pasquale , Dharmapuri Sridhar , Nuzzo Maurizio , Maldini Daniele , Iezzi Manuela , Cavallo Federica , Musiani Piero , Forni Guido , Monaci Paolo
Published: August 2004

The protective efficacy of xenogeneic vaccination with DNA encoding the HER2 oncogene was evaluated in BALB/c mice transgenic for the transforming form of the neu oncogene, which spontaneously develops carcinomas in all mammary glands. Intramuscular injection of either plasmid DNA followed by electrical stimulation (pVIJ‐HER2 with ES) or an adenoviral vector (Ad5‐HER2), both expressing the HER2 oncogene, was tested. Immunization using pVIJ‐HER2 with ES elicited a cell‐mediated response that was much lower than that elicited by the immunization with Ad5‐HER2, as measured by the frequency of IFN‐γ‐secreting spleen cells. The dominant T‐cell epitope of the HER2 protein product (p185) in the BALB/c (H‐2d) genetic background was identified. While the T‐cell response elicited was only partially crossreactive with the corresponding rat epitopes because of sequence variations (89% similarity), a cytotoxic T‐lymphocyte activity against the rat immunodominant epitope was also evident. The Ad5‐HER2 vaccination induced also antibodies against p185, which crossreacted with the rat protein homolog. Both T‐ and B‐cell responses slowly declined with time. Vaccination with Ad5‐HER2 at 6 and 9 weeks of age delayed incidence and reduced multiplicity of tumors in neu transgenic mice.
CD95L mediates tumor counterattack in vitro but induces neutrophil‐independent tumor rejection in vivo
Authors: Igney Frederik H. , Behrens Christian K. , Krammer Peter H.
Published: August 2004

Many tumors express CD95L (CD178, FasL, APO‐1L) and may thus kill tumor‐infiltrating lymphocytes, a phenomenon called tumor counterattack. However, presently it is not clear whether tumor counterattack is a relevant immune escape mechanism. To characterize the effect of CD95L expression of tumor cells on tumor‐specific T cells, we established an in vitro system with TCR tg T cells and a model tumor antigen. Preactivated antitumor T cells were able to kill CD95L− and CD95L+ tumor cells. CD95L+ tumor cells killed activated T cells in vitro and inhibited the expansion of cytotoxic antitumor T cells in mixed lymphocyte tumor reactions. In vivo CD95L expression led to delayed tumor growth or complete tumor rejection. Neutrophils were not responsible for the delayed growth of the CD95L+ tumors tested. In mice with neutrophils deficient for important cytotoxicity mechanisms (p47phox−/− or iNOS−/− mice), CD95L+ tumors grew similarly as in wild‐type mice. Incidence and growth rate of CD95L+ tumors in mice injected with a neutrophil‐depleting or an isotype control antibody was the same. In CD95‐deficient lpr mice, tumor growth was not altered as compared to wild‐type mice. Taken together, CD95L mediated tumor counterattack in vitro, but led to neutrophil‐independent tumor rejection in vivo.
Influence of glutathione S‐transferase pi and p53 expression on tumor frequency and spectrum in mice
Authors: Gate Laurent , Majumdar Rajrupa S. , Lunk Alexandra , Tew Kenneth D.
Published: August 2004

The role of glutathione S‐transferase π (GSTπ) in tumor development has been previously suggested; however the exact function of this enzyme in carcinogenesis remains unclear. GSTπ has been identified as a modulator of cell signaling by interacting with and inhibiting c‐Jun N‐terminal kinase (JNK). This kinase has been in turn described as a regulator of p53 stability and transcriptional activity. To study the possible interaction between GSTπ and p53, we crossed GSTπ‐deficient animals with p53−/− mice. Double knock out animals were viable but developed tumors within 6 months of age; the life span of these animals was however similar to that of GSTπ+/−/p53−/− and GSTπ+/+/p53−/−. Mice heterozygous for p53 lived significantly longer than the p53−/− animals and developed tumors much later, and the expression of GSTπ did not significantly modify the life span of the animals. In contrast, in a wild‐type p53 background, GSTπ−/− mice developed tumors with a significantly higher frequency than heterozygous and wild‐type animals with a median tumor free life span 20 weeks shorter. In addition, in p53+/+ background, one third of the GSTπ−/− animals developed lung adenomas, while less than 10% of GSTπ+/− and GSTπ+/+ presented such tumors. GSTπ expression did not alter the expression of tumorigenesis markers such as COX‐2 or ornithine decarboxylase in response to phorbol ester. Furthermore, GSTπ‐deficient mouse embryo fibroblasts were more sensitive to H2O2‐induced apoptosis. P53−/− cells, independent of GSTπ status, were more sensitive to UV and other DNA damaging agents than their wild‐type counterparts. These results suggest that GSTπ may play a protective role in the development of spontaneous tumors.
Mutant BRCA1 alleles transmission: Different approaches and different biases
Authors: de la Hoya Miguel , Meijers‐Heijboer Hanne , Fernández Juan Manuel , Díez Orland , Osorio Ana , Alonso Carmen , van Leeuwen Inge , Díaz‐Rubio Eduardo , Cornelisse Cees , Benítez Javier , Devilee Peter , Caldés Trinidad
Published: August 2004

Recently, a non‐random transmission of BRCA1 mutant alleles to the off‐spring of carriers has been suggested. If confirmed by further studies, the finding will have important implications both for risk assessment and the understanding of BRCA1 biology. However, these kind of analysis are not easy to perform due to several ascertainment biases inherent to the identification of BRCA1 carriers. In this report we propose different approaches to overcome such biases and we present additional data supporting a non‐random transmission of BRCA1 mutant alleles.
Lack of prognostic implications of HER‐2/neu abnormalities in 345 stage I nonsmall cell carcinomas (NSCLC) and 207 stage I–III neuroendocrine tumours (NET) of the lung
Authors: Pelosi Giuseppe , Del Curto Barbara , Dell'Orto Patrizia , Pasini Felice , Veronesi Giulia , Spaggiari Lorenzo , Maisonneuve Patrick , Iannucci Antonio , Terzi Alberto , Lonardoni Alessandro , Viale Giuseppe
Published: August 2004

HER‐2/neu oncogene activation by either gene amplification and/or protein overexpression has been documented in several human malignancies. Irrespective of protein overexpression, HER‐2/neu gene amplification is rare in lung cancer and studies on its prevalence and clinicopathological implications in early stage nonsmall cell lung cancer (NCSLC) and neuroendocrine tumours (NET) of the lung are lacking. We evaluated HER‐2/neu abnormalities in 345 Stage I NSCLC and 207 Stage I‐III NET of the lung of all the diverse histological types, by using immunohistochemistry and fluorescent in situ hybridization in selected cases. Overall, HER‐2/neu immunoreactivity was detected in 23% of 345 NSCLC and in 7% of 207 NET. Gene amplification was seen in only 7 (7.4%) of the immunoreactive tumours, with high‐level amplification (HER‐2/neu gene to chromosome 17 ratio > 4.0) in 3 adenocarcinomas, 1 squamous‐cell carcinoma and 1 large‐cell neuroendocrine carcinoma (LCNEC), and low‐level amplification (HER‐2/neu gene to chromosome 17 ratio from 2.0 to 4.0) in 1 squamous‐cell carcinoma and 1 LCNEC. None of tested carcinoids and SCLC showed gene amplification. All but 1 gene amplified case exhibited 2+ or 3+ membrane labeling. No relationship was found between gene amplification or protein overexpression and patients' survival or other clinicopathological variables. HER‐2/neu gene amplification and protein overexpression are not closely correlated in lung carcinomas and do not bear any prognostic implication. Among neuroendocrine tumours, LCNEC show a slightly higher prevalence of either HER‐2/neu gene amplification or protein overexpression.
A recombinant humanized anti‐insulin‐like growth factor receptor type I antibody (h7C10) enhances the antitumor activity of vinorelbine and anti‐epidermal growth factor receptor therapy against human cancer xenografts
Authors: Goetsch Liliane , Gonzalez Alexandra , Leger Olivier , Beck Alain , Pauwels Petrus J. , Haeuw Jean François , Corvaia Nathalie
Published: August 2004

Interaction of insulin‐like growth factor receptor I (IGF‐IR) with its ligands has been reported to induce cell proliferation, transformation and blockade of cell apoptotic functions. IGF‐IR is overexpressed on numerous tumor cell types and its blockade could be of importance for anti‐cancer therapy. We have generated a humanized anti‐IGF‐IR antibody h7C10 that blocksin vitroIGF‐I and IGF‐II‐induced cell proliferation of MCF‐7 breast cancer cells. Analysis of the IGF‐I transduction cascade demonstrated that the humanized anti‐IGF‐IR antibody and its murine parental form block IGF‐I‐induced tyrosine phosphorylation, both its β‐chain and IRS‐1 tyrosine phosphorylation. This presumably leads to cell cycle arrest and, consequently, growth inhibition. Treatment of nude mice bearing either human breast cancer cells (MCF‐7) or non small lung cancer cells (A549) with h7C10, or its murine parental form 7C10, inhibited significantly tumor growth. An almost complete inhibition of A549 tumor growth was observed when mice were treated with the anti‐IGF‐IR antibody combined with either a chemotherapeutic agent, Vinorelbine or an anti‐epidermal growth factor receptor (EGFR) antibody, 225. Combined therapy prolonged significantly the life span of mice in an orthotopicin vivomodel of A549; the combination of the anti‐IGF‐IR antibody with an anti‐EGFR antibody was superior to the Vinorelbine combination. The present results indicate that the humanized anti‐IGF‐IR antibody h7C10 has a great potential for cancer therapy when combined with either a chemotherapeutic agent or an antibody that targets other growth factor receptors, such as the epidermal growth factor receptor.
Overexpression of a hybrid gene consisting of the amino‐terminal fragment of urokinase and carboxyl‐terminal domain of bikunin suppresses invasion and migration of human ovarian cancer cells in vitro
Authors: Takei Yuji , Mizukami Hiroaki , Saga Yasushi , Kobayashi Hiroshi , Suzuki Mika , Matsushita Takashi , Ozawa Keiya , Suzuki Mitsuaki
Published: August 2004

A Kunitz‐type protease inhibitor, bikunin, is known to suppress the invasion and metastasis of cancer cells. HI8, a carboxyl‐terminal domain of bikunin, is an active site of this glycoprotein. To increase its affinity for cancer cells, we constructed a chimeric gene, ATF‐HI8, and investigated the anti‐invasive and anti‐migratory activity of ATF‐HI8 on ovarian cancer cells. ATF‐HI8‐expressing plasmid and ATF‐expressing plasmid were introduced into the highly invasive and metastatic ovarian cancer cell line HRA. The properties of the established cell line (HRA/ATF‐HI8) were compared to those of the HRA/ATF and the HRA/luciferase (HRA/LUC, control) cell lines in terms of cell proliferation, invasion and migration. As a result, (i) there were no differences in cell proliferation between HRA/ATF‐HI8 and HRA/LUC; (ii) the invasion and migration of HRA/ATF‐HI8 cells were significantly inhibited compared to those of HRA/LUC cells; (iii) the migration, but not the invasion, of HRA/ATF cells was significantly inhibited compared to that of HRA/LUC. These results indicate that the overexpression of ATF‐HI8 inhibits the invasion and migration of ovarian cancer cells without affecting cell proliferation and suggest that HI8 is involved in the anti‐invasive and the anti‐migratory activities, and the addition of ATF brought about the increase in the anti‐migratory activity of HI8. The above findings suggest the applicability of therapeutic strategies targeting the inhibition of peritoneal invasion and dissemination of ovarian cancer by the use of the chimeric gene ATF‐HI8.
Epidermal growth factor receptor (EGFR) downstream molecules as response predictive markers for gefitinib (Iressa®, ZD1839) in chemotherapy‐resistant non‐small cell lung cancer
Authors: Han Sae‐Won , Hwang Pil Gyu , Chung Doo Hyun , Kim Dong‐Wan , Im Seock‐Ah , Kim Young Tae , Kim Tae‐You , Heo Dae Seog , Bang Yung‐Jue , Kim Noe Kyeong
Published: August 2004

Gefitinib has shown meaningful antitumor activity with tolerable toxicity in chemotherapy‐refractory NSCLC in previous studies. Moreover, EGFR expression failed to show a correlation with response. In an attempt to identify predictive markers of response, we have investigated the tumoral expression of key signaling molecules of EGFR (EGFR, p‐EGFR, p‐Akt, p‐Erk, p‐STAT3) by immunohistochemistry and analyzed their correlations with response. Of 65 patients who received gefitinib (250 mg/day) for chemotherapy‐refractory NSCLC, there were 14 partial responses (21.5%), 21 stable diseases (32.3%) and 21 progressive diseases (32.3%). Median durations of overall survival and time to progression were 6.7 months and 2.8 months, respectively. Immunohistochemistry was performed in 34 patients with evaluable tissue specimens. EGFR was overexpressed (2+ or 3+) in 32.4% and p‐EGFR was positive in 26.5%. The expressions of p‐Akt, p‐Erk and p‐STAT3 were positive (1+ or 2+) in 50%, 38.2% and 79.4%, respectively. The EGFR expression was not correlated with p‐EGFR or the downstream molecules. EGFR or p‐EGFR status did not correlate with response. Positive expression of p‐Erk was significantly associated with poor response (38.1% in −, 14.3% in 1+, 0% in 2+; p = 0.046). Furthermore, tumors with positive p‐Akt and negative p‐Erk nuclear expression exhibited the best response (60%), whereas there was no response in the opposite [p‐Akt (−), p‐Erk (+)] cases. Intense nuclear staining of p‐Akt (2+) was associated with prolonged TTP (HR 0.25, 95% confidence interval [CI] 0.08–0.79, p = 0.018) and OS (HR 0.16, 95% CI 0.04–0.62, p = 0.008). These results support the assumption that gefitinib responsiveness might be predicted by activated EGFR downstream molecules such as p‐Akt and p‐Erk.
Explaining differences in incidence rates of pleural mesothelioma between Sweden and the Netherlands
Authors: Burdorf Alex , Järvholm Bengt , Englund Anders
Published: August 2004

In recent years in several countries a deceleration or leveling off of pleural mesothelioma rates has been observed. The impact of asbestos used was analysed by comparing a country with a relative modest incidence rate of mesothelioma (Sweden) and an early response to asbestos use with a country with one of the highest incidence rates of mesothelioma in Western Europe (The Netherlands). In Sweden the Cancer Register provided information on the annual incidence of pleural mesothelioma, whereas in The Netherlands mortality data were provided by Statistics Netherlands for the period 1969–2001. In The Netherlands among men the incidence rate was consistently higher (1.5–2 times) than in Sweden, whereas among women similar rates were observed. Assuming that none of the female cases was caused by occupational exposure to asbestos, minimum estimates of the etiologic fraction for occupational exposure to asbestos in men would be 82% in Sweden and 92% in The Netherlands. Possible explanations for the consistently higher incidence rates in the Netherlands than in Sweden include differences in exposure levels, the proportion of exposed subjects in the workforce and types of asbestos fibres used. Measures to decrease the exposure to asbestos seem to have decreased the risk of pleural mesothelioma in both countries among age groups below 60 years. This effect will result in a leveling off of the increase in pleural mesothelioma in both countries in the next decade.
E5 protein of human papillomavirus type 16 selectively downregulates surface HLA class I
Authors: Ashrafi G. Hossein , Haghshenas Mohammad R. , Marchetti Barbara , O'Brien Philippa M. , Campo M. Saveria
Published: August 2004

Papillomaviruses have evolved mechanisms that result in escape from host immune surveillance. The E5 protein is expressed early in papillomavirus infection in the deep layers of the infected epithelium. It is localized to the Golgi apparatus (GA) and endoplasmic reticulum. The E5 protein of bovine papillomavirus (BPV) impairs the synthesis and stability of major histocompatibility (MHC) class I complexes and prevents their transport to the cell surface due to retention in the GA. Here we show that human papillomavirus type 16 (HPV‐16) E5 also causes the retention of MHC (HLA) class I complexes in the GA and impedes their transport to the cell surface, which is rescued by treatment with interferon. Unlike BPV E5, HPV‐16 E5 does not affect the synthesis of HLA class I heavy chains or the expression of the transporter associated with antigen processing TAP. These results show that downregulation of surface MHC class I molecules is common to both BPV and HPV E5 proteins. Moreover, we determined that HPV‐16 E5 downregulates surface expression of HLA‐A and HLA‐B, which present viral peptides to MHC class I‐restricted cytotoxic T lymphocytes (CTLs), but not the natural killer (NK) cell inhibitory ligands HLA‐C and HLA‐E. Selective downregulation of cell surface HLA class I molecules may allow the virus to establish infection by avoiding immune clearance of virus‐infected cells by both CTLs and NK cells.
Intracellular LPS inhibits the activity of potassium channels and fails to activate NFκB in human macrophages
Authors: Gerth Anja , Grosche Jens , Nieber Karen , Hauschildt Sunna
Published: August 2004

Although much has been learned about signal transduction mechanisms and binding proteins involved in lipopolysaccharides (LPS)‐induced activation of monocytes/macrophages, little is known about the ability of internalized LPS to activate cells. To approach this question we either exposed macrophages to LPS or microinjected the cells with LPS before studying early cellular events associated with LPS‐mediated macrophage activation. We measured membrane currents and translocation of NFκB to the nucleus. Using the whole‐cell patch clamp technique ion channels were analyzed and characterized as K+ sensitive inward and outward currents. Exogenous LPS was shown to increase the voltage‐dependent outward current whereas the voltage‐dependent inward current was unaffected. However when cells were microinjected with LPS both inward and outward current were completely abolished. The presence of LPS within the cells did not prevent them to perform phagocytosis or to respond to fMLP with an appropriate increase in [Ca2+]i. The immunocytological detection of NFκB p65 translocation revealed that exogenous LPS led to the nuclear localization of the p65 subunit of NFκB, whereas only the cytoplasmic localization of p65 was observed following microinjection of LPS. These data show that one major process in macrophage activation, the NFκB dependent transcription of a number of genes encoding for many inflammatory mediators cannot be induced by intracellular LPS but requires the interaction of LPS with external membrane components. However intracellular LPS causes a drastic decrease in potassium currents which by keeping the cell membrane at a depolarized potential may result in changed biological answers of these cells. © 2004 Wiley‐Liss, Inc.
Sustained Wnt protein expression in chondral constructs from mesenchymal stem cells
Authors: Nishioka Katsuhiro , Dennis James E. , Gao Jizong , Goldberg Victor M. , Caplan Arnold I.
Published: August 2004

Wnt genes encode a number of secreted glycoproteins which are closely associated with the cell surface and the extracellular matrix. Recently, members of Wnt family have been implicated in regulating chondrocyte differentiation, but their roles in the chondrogenic process are not fully understood. To contribute to an understanding of the roles of Wnts during chondrogenesis, we have analysed the spatiotemporal expression patterns of Wnt using in vitro models for chondrogenesis of human bone marrow‐derived mesenchymal stem cells (hMSCs). In chondrogenic aggregate culture system, RT‐PCR analysis revealed expression of Wnt5a and Wnt4 during late chondrogenesis (days 10 and 15). Immunohistochemical analysis showed widespread distribution of Wnt5a and Wnt4 throughout the aggregates at this late phase of culture (days 14 and 21). In addition, in this aggregate culture system, immunohistochemical staining of Wnt4 and Wnt5a showed similar spatiotemporal expression patterns to that of type II collagen or type X collagen. To confirm the results obtained by immunostaining, the specificity of the anti‐Wnt4 or anti‐Wnt5a antibody was assessed by Western blot analysis. Of Wnt4 and Wnt5a, only Wnt5a was immunodetectable by Western blot analysis. Western blot analysis showed that Wnt5a was expressed as two different molecular weight forms of 40 and 44 kDa. Treatment with PNGase F, which removes N‐linked oligosaccharides, revealed that the mass difference between these two forms could be accounted for by the N‐glycosylation status of the protein. When hMSCs were seeded on a porous gelatin sponge, immunolocalization studies showed that type II collagen and type X collagen were detected particularly at the periphery at day 7 of culture. In contrast, Wnt4 and Wnt5a showed even distribution throughout the hMSC/gelatin sponge constructs. Their different spatial expression patterns suggest that Wnt4 and Wnt5a proteins are not functionally linked to type II collagen and type X collagen synthesis in in vitro chondrogenic models of hMSCs. © 2004 Wiley‐Liss, Inc.
Radiosynthesis of [11C]docetaxel
Authors: van Tilburg E. W. , Franssen E. J. F. , van der Hoeven J. J. M. , van der Meij M. , Elshove D. , Lammertsma A. A. , Windhorst A. D.
Published: August 2004

Docetaxel (Taxotere®) is an accepted chemotherapeutic agent for the treatment of breast cancer and non‐small cell lung cancers. A potential means of predicting response is measuring tumor uptake of [11C]docetaxel using Positron Emission Tomography (PET). The synthetic approach to introduce the 11C isotope in the 2‐benzoyl moiety of docetaxel unfortunately was unsuccessful. The radiosynthesis of [11C]docetaxel (6b, Scheme 1), with the 11C isotope in the BOC moiety, was however, successful using a second synthetic approach. It started with the reaction of [11C]tert‐butanol with 1,2,2,2‐tetrachloroethyl chloroformate to give [11C]tert‐butyl‐l,2,2,2‐tetrachloroethyl carbonate in a good overall yield (62±9%). In the final step, the [11C]tert‐butoxycarbonylation of the free amine of docetaxel gave [11C]docetaxel 6b in a satisfactory decay corrected yield of 10±1% (from [11C]CO2). Copyright © 2004 John Wiley & Sons, Ltd.
Ochre decor and cinnabar residues in Neolithic pottery from Vinča, Serbia
Authors: Mioč U. B. , Colomban Ph. , Sagon G. , Stojanović M. , Rosić A.
Published: August 2004

The prehistoric site of Vinča, on the right bank of the River Danube, the territory of the City of Belgrade, first excavated by M. Vasić in 1931–34, provides Neolithic pottery dating back to 5200–4200 BC. Shards excavated in 1998 (Serbian Academy of Sciences and Arts) have a yellow coating on the external (convex) and red deposit on the internal (concave) side. Raman, IR and x‐ray identification proved that the yellow‐to‐red decor deposited on the external faces of pottery is made of ochre, a mixture of hematite, quartz and phyllosilicates. The red deposit found on some internal surfaces of the pottery consists of cinnabar (HgS) with some quartz and phyllosilicates. This indicates that cinnabar was not used for decor but for other purposes than and prepared or stored in ceramic utensils. A comparison is made with mercury ore from the Šuplja Stena mine located ∼20 km from Vinča village. Copyright © 2004 John Wiley & Sons, Ltd.
Vibrational spectroscopic characteristics of phthalocyanine and naphthalocyanine in sandwich‐type phthalocyaninato and porphyrinato rare earth complexes. Part 11—Raman spectroscopic characteristics of phthalocyanine in mixed [tetrakis(4‐chlorophenyl)porphyrinato](phthalocyaninato) rare earth double‐deckers
Authors: Lu Fanli , Rintoul Llew , Sun Xuan , Arnold Dennis P. , Zhang Xianxi , Jiang Jianzhuang
Published: August 2004

Raman spectroscopic data in the range 500–1800 cm−1 for a series of 15 mixed [tetrakis(4‐chlorophenyl)porphyrinato](phthalocyaninato) double‐decker complexes with tervalent rare earths MIII(TClPP)(Pc) (M = Y, LaLu except Ce and Pm) and intermediate‐valent cerium Ce(TClPP)(Pc) were collected using laser excitation sources emitting at 632.8 and 785 nm. Comparison with the Raman spectra of corresponding bis(phthalocyaninato) rare earths reveals that the Raman characteristics of mixed‐ring double‐deckers are dominated by the phthalocyaninato‐metal fragment M(Pc) and phthalocyanine exists as the monoradical anion Pc•− in MIII(TClPP)(Pc) and the dianion Pc2− in Ce(TClPP)(Pc), respectively. With excitation at 632.8 nm, which is in close resonance with the main Q absorption band of the phthalocyanine ligand, a typical Raman marker band of the monoanion radical Pc•− was observed at 1512–1519 cm−1 as a weak band resulting from the coupling of pyrrole CC and aza CN stretchings. For Ce(TClPP)(Pc), a strong band at 1492 cm−1 with contributions from both pyrrole CC and aza CN stretchings and also isoindole stretching was the marker Raman band of Pc2−. When excited with laser radiation of 785 nm, which is far away from the main Q absorption band of the phthalocyanine ligand, the marker Raman band of Pc•− in MIII(TClPP)(Pc) is observed at 1495–1514 cm−1 and of Pc2− in Ce(TClPP)(Pc) at 1511 cm−1 as a strong band. Copyright © 2004 John Wiley & Sons, Ltd.
Surface‐enhanced Raman scattering study of the adsorption of the anthraquinone pigment alizarin on Ag nanoparticles
Authors: Cañamares M. V. , Garcia‐Ramos J. V. , Domingo C. , Sanchez‐Cortes S.
Published: August 2004

FT‐Raman and surface‐enhanced Raman scattering (SERS) spectroscopy were applied in the vibrational characterization and study of the adsorption and acidity behavior of the highly fluorescent anthraquinone dye alizarin on Ag colloids prepared by chemical reduction with hydroxylamine hydrochloride. The SERS spectra were obtained at different conditions of pH, excitation wavelength and pigment concentration in order to deduce the adsorption mechanism of this molecule. On the basis of the results found we propose an adsorption model for alizarin, which has a different acidic behavior on the metal surface to that in solution. On the metal the deprotonation order of the OH groups changes with respect to the aqueous solution, the OH in position 1 being the first to be ionized instead of that in position 2 as occurs in solution. The two main alizarin forms identified on the metal surface correspond to the mono‐ and dianionic alizarin species. Copyright © 2004 John Wiley & Sons, Ltd.
Time‐resolved resonance Raman and density functional theory investigation of the 4‐biphenylnitrene dimerization reaction
Authors: Chan Pik Ying , Ong Shing Yau , But Tracy Yuen Sze , Phillips David Lee
Published: August 2004

This paper reports time‐resolved resonance Raman spectra for the triplet 4‐biphenylnitrene and its reaction to form an azo dimer product (bis‐biphenyl‐4‐yldiazene) in acetonitrile solution. The properties of the triplet 4‐biphenylnitrenes were examined based on the results of UBPW91/cc‐PVDZ computations. 4‐Biphenylnitrene and its azo dimer product were also compared with results previously reported for the 4‐methoxyphenylnitrene and its azo dimer product, respectively. The substitution effect of p‐phenyl and p‐methoxy moieties on the structure and properties of arylnitrenes and azobenzenes are briefly discussed. Copyright © 2004 John Wiley & Sons, Ltd.
NIR‐FT Raman and FT‐IR spectral studies and ab initio calculations of the anti‐cancer drug combretastatin‐A4
Authors: Binoy J. , Abraham Jose P. , Joe I. Hubert , Jayakumar V. S. , Pettit G. R. , Nielsen O. F.
Published: August 2004

Combretastatin‐A4, a potential anti‐cancer drug in advanced preclinical development, is extracted from the medicinal plant Combretum caffrum. NIR‐FT Raman and FT‐IR spectral studies were carried out and ab initio calculations were performed at the HF/6–31G(d) level to derive equilibrium geometry and vibrational wavenumbers and intensities. The vibrational analysis shows that the molecule exhibits similar geometric behavior to cis‐stilbene, and has undergone steric repulsion resulting in phenyl ring twisting with respect to the ethylenic plane. The spectral behavior of the methoxy groups is influenced by the oxygen lone pair–aryl pz orbital conjugation and back‐donation. Chelation of the OH group with the oxygen lone pair of the methoxy group produces separate bands for free OH and associated OH vibrations. Copyright © 2004 John Wiley & Sons, Ltd.
Volatile aldehyde emissions from heated cooking oils
Authors: Fullana Andres , Carbonell‐Barrachina Ángel A , Sidhu Sukh
Published: August 2004

Low molecular weight aldehydes (LMWAs) formed during the heating of frying media (triglycerides) were adsorbed onto tenax and analyzed by GC‐MS after thermal desorption. Six alkanals (C5 to C10), seven 2‐alkenals (C5 to C11) and 3 alkadienals (C7, C9 and C10) were found in the fumes of canola oil (control), extra virgin olive oil, and refined olive oil, heated at 180 and 240 °C. The emission rates of these aldehydes depended on the heating temperature. Frying in any type of olive oil, independently of its commercial category, will effectively decrease the emission of volatile aldehydes at temperatures below the smoking point. Thus, using the cheaper olive oil for deep‐frying purposes will not affect aldehyde emissions. This is important since olive oil is usually used for deep‐frying operations while extra virgin olive oil is used as salad dressing in Spain. The mixture of refined olive oil with some virgin olive oil is the most acceptable type of olive oil in non‐Mediterranean countries due to its milder flavor. However, if higher temperatures are needed the use of canola oil is more advisable due to its higher smoke point. Copyright © 2004 Society of Chemical Industry
Functionality of native and denatured cashew nut kernel protein isolates at isoelectric pH as a function of salt concentration
Authors: Bora Pushkar Singh , Neto Vicente Queiroga
Published: August 2004

The reduced solubility of proteins near the isoelectric pH limits their use in food formulations whose pH lies in the range 5.0–6.0 because of poor functionality. In the present study, the effect of salt on the functionality of native and denatured cashew nut kernel protein isolates at the isoelectric pH was investigated. Both isolates showed improvement in their functional properties, but the improvement was greater for the denatured protein isolate. The solubilities of denatured and native protein isolates at the isoelectric pH increased from 26.4 g l−1 and 64 g l−1, respectively, without salt to maxima of 363 and 308 g l−1, respectively, at 0.75 M salt concentration. The water binding capacity of the isolates increased with increase in NaCl concentration from 1.70 ml g−1 to 1.77, 1.82, 1.92 and 2.2 ml g−1 for denatured protein isolate and from 1.45 ml g−1 to 1.65, 1.69, 1.82 and 1.97 ml g−1 for native protein isolate at 0.25, 0.50, 0.75 and 1.0 M salt concentrations, respectively. When the properties of the isolates in 0.75 M NaCl solutions were compared with those in salt‐free water there were 15% and 116% increases in emulsifying capacity, 40‐fold and 45‐fold increases in emulsifying activity and 4.6‐fold and 40‐fold increases in emulsion stability for native and denatured protein isolates, respectively, whilst the corresponding foaming capacities increased from 4 to 5.5 and 0 to 8.9 ml g−1 protein. Statistically, no difference in the foaming capacity of either of the isolates was observed above 0.5 M NaCl. The foam stability also exhibited similar behaviour. Copyright © 2004 Society of Chemical Industry