bluuebunny/crossref_metadata_2025 huggingface dataset:
Analysis of production, purification, and cytolytic potential of bi‐specific antibodies reactive with ovarian‐carcinoma‐associated antigens and the T‐cell antigen CD3
Authors: van Ravenswaay Claasen Hedda H. , van de Griend René J. , Mezzanzanica Delia , Bolhuis Reinder L. H. , Warnaar Sven O. , Fleuren Gert Jan
Published: February 2007

OV‐TL 3 and MOvl8 MAbs, due to their restricted specificity, have been successfully used to visualize ovarian cancer in patients and might therefore be used to develop therapies for ovarian cancer. The bi‐specific MAbs αT3/OC2 and αOC/TR (both being combinations of MOvl8 and aCD3) have been shown to lyse ovarian tumor cells in vitro. To evaluate the relative merits of MOvl8/CD3 and OV‐TL 3/CD3, the present study was undertaken in which the bi‐specific MAbs αT3/OC2 and αOC/TR, and a newly developed bi‐specific MAb, OV‐TL 3/CD3, were highly purified and compared for specificity, stability, purification and cytolytic potential. The dual specificity of the hybrid‐hybridoma supernatants was analyzed by immunohistochemistry, and by testing bi‐specific MAb‐mediated cytotoxicity against relevant target cells in the presence of effector cells. Stability testing of bi‐specific MAb‐producing hybrid‐hybridomas showed that, after sub‐cloning, clones stably produced up to 40% bi‐specific MAb even after prolonged in vitro culture. The purification of the bi‐specific fractions was performed with protein A and by ion‐exchange high‐pressure liquid chromatography, depending on the sub‐class combination of the bi‐specific MAb. The purified bi‐specific MAbs were tested for their ability to mediate target‐cell lysis with the use of cytotoxic T‐cell clones and activated peripheral‐blood lymphocytes. The purified αT3/OC2, αOC/TR, and OV‐TL 3/CD3 were all able to mediate highly specific lysis of various ovarian‐carcinoma cell lines. No correlation was found between the level of antigen expression and bi‐specific MAb‐mediated cytolysis. © 1993 Wiley‐Liss, Inc.
Invasive squamous‐cell cervical carcinoma and combined oral contraceptives: Results from a multinational study
Authors: Berry Geoffrey , Maclennan Robert , Shearman Rodney , Jelihovsky Tatiana , Booth Joan Cooper , Molina Ramiro , Martinez Luis , Salas Oriana , Dabancens Alfredo , Zhiheng Chen , Yun Tao , Wei Hu Yong , Cuadros Alvaro , Aristizabal Nubia , Modan Baruch , Ron Elaine , Alfandary Esther , Mati J. G. , Kenya Patrick , Kungu Alfred , Gatei D. , Cuevas Hector Rodriguez , Salazar Socorro Benavides , Palet Antonio , Ontiveros Patricia , Ibeziako Pat A. , Junaid T. A. , Aghediuno P. , Abioye A. A. , Apelo Ruben A. , De la Cruz Julietta R. , Baens Jose , Canlas Benjamin D. , Silpisornkosol Suporn , Pardthaisong Tieng , Charoeniam Viruch , Theetranont Choti , Boosiri Banpot , Chutivongse Supawat , Virutamasen Pramuan , Wongsrichanalai Chansuda , Sindhavananda Sermsri , Koetsawang Suporn , Rachawat Duangdao , Koetsawang Amorn , Riotton Gustave , Christopherson William M. , Melnick Joseph L. , Adam Ervin , Thomas David B. , Ray Roberta M. , Noonan Elizabeth A. , Stanford Janet L. , Rosenblatt Karin A. , Holck Susan , Meirik Olav , Farley Timothy M. M. , Thomas David B. , Ray Roberta M.
Published: February 2007

Data from a hospital‐based case‐control study collected in 11 participating centers in 9 countries were analyzed to determine whether use of combined oral contraceptives alters risk of invasive squamous‐cell cervical cancer. Information on prior use of oral contraceptives, screening for cervical cancer, and suspected risk factors for this disease were ascertained from interviews of 2361 cases and 13,644 controls. A history of smoking and anal and genital warts was obtained, and blood specimens were collected for measurement of antibodies against herpes simplex and cytomegalo viruses, from selected sub‐sets of these women, as was a sexual history from interviews of husbands. The relative risk of invasive squamous‐cell cervical carcinoma was estimated to be 1.31, with a 95% confidence interval that excluded one, in women who ever used combined oral contraceptives. Risk of this disease increased significantly with duration of use after 4 to 5 years from first exposure, and declined with the passage of time after cessation of use to that of non‐users in about 8 years. No sources of bias or confounding were identified that offered plausible explanations for these findings. The strength of these results, and their consistency with those from other studies, suggest that a causal relationship may exist between use of combined oral contraceptives and squamous‐cell cervical carcinoma. Women who have used these products for 4 or more years, and who most recently used them within the past 8 years, should receive high priority for cervical cytologic screening.
Photodynamic therapy with m‐tetrahydroxyphenylchlorin in vivo: Optimization of the therapeutic index
Authors: Ris Hans‐Beat , Altermatt Hans J. , Stewart Charles M , Schaffner Thomas , Wang Qiang , Lim Chang K. , Bonnett Raymond , Althaus Ulrich
Published: February 2007

The therapeutic index of meta‐tetrahydroxyphenylchlorin‐mediated photodynamic therapy (mTHPC‐PDT) was assessed in BALB/c nude mice bearing human malignant mesothelioma xenografts. Equal doses of 650 nm laser light were delivered to the tumour and to an equal‐sized area of the hind leg (control site) after i.p. administration of mTHPC. Twenty‐one groups of 6 animals each were treated under various drug‐light conditions and at drug‐light intervals ranging from 4 hr to 6 days. After light delivery the extent of tumour necrosis and the depth of alterations in normal tissue were assessed by light microscopy of standardized histological sections. A therapeutic index (T1) of mTHPC‐PDT was defined as the cross‐sectional area of tumour necrosis per depth of visible tissue injury at the control site. This T1 was strongly related to the conditions of treatment. In particular, it was increased by prolonging the drug‐light interval up to 5 days and by increasing the dose of light for any dose of drug. The most profound increase of T1 was obtained by increasing the intensity of light administered at the chosen interval while reducing the dose of drug. Our findings suggest that threshold conditions operate in PDT and have important implications for clinical application of the treatment.
Variable effects of sodium butyrate on the expression and function of the MDR 1 (P‐glycoprotein) gene in colon carcinoma cell lines
Authors: Frommel Thomas O. , Coon John S. , Tsuruo Takashi , Roninson Igor B.
Published: February 2007

Expression of the MDRI (P‐glycoprotein) gene confers resistance to several classes of chemotherapeutic drugs (multi‐drug resistance). Colon carcinomas frequently express high levels of MDRI mRNA and P‐glycoprotein, presumably reflecting the origin of these tumors from MDRI ‐expressing normal colonic cells. In 4 colon carcinoma cell lines (SW 620, HCT‐15, DLD‐I, LS 180), MDRI expression was reported in an earlier study to be elevated after exposure to a differentiating agent, sodium butyrate (NaB). In one of these cell lines (SW 620), increased MDR I expression reportedly was not accompanied by a decrease in the accumulation or cytotoxicity of vinblastine, a P‐glycoprotein‐transported drug, suggesting a possible functional abnormality of NaB‐induced P‐glycoprotein. We have re‐examined the effect of NaB on MDRI /P‐glycoprotein expression and function in the same colon carcinoma cell lines. NaB treatment induced differentiation‐related changes and increased expression of MDRI mRNA in all 4 cell lines. A major increase in MDRI mRNA and P‐glycoprotein expression was observed in only one line, SW 620. This increase, however, was accompanied by a commensurate increase in the activity of P‐glycoprotein, indicating that the induced protein was fully functional. NaB treatment caused a relatively minor increase in MDRI mRNA expressed in the other 3 cell lines. Two of these lines showed a detectable increase in the P‐glycoprotein expression and function, but in the third line (LS 180) P‐glycoprotein was undetectable either before or after exposure to NaB. The magnitude of MDRI induction by NaB showed no apparent correlation with differentiation‐related changes induced by this agent.
A polymerase‐chain‐reaction assay for the specific identification of transcripts encoded by individual carcinoembryonic antigen (CEA)‐gene‐family members
Authors: Thompson J. , Mössinger S. , Reichardt V. , Engels U. , Beauchemin N. , Kommoss F. , von Kleist S. , Zimmermann W.
Published: February 2007

Carcinoembryonic antigen (CEA) is a tumor marker that belongs to a family of closely related molecules with variable expression patterns. We have developed sets of oligonucleotide primers for the specific amplification of transcripts from individual CEA‐family members using the reverse transcriptase/ polymerase chain reaction (RT/PCR). Specific primer sets were designed for CEA, non‐specific cross‐reacting antigen (NCA), biliary glycoprotein (BGP), carcinoembryonic antigen gene‐family members 1, 6 and 7 (CGMI, CGM6 and CGM7), and one set for all pregnancy‐specific glycoprotein (PSG) transcripts. Primers were first tested for their specificity against individual cDNA clones and product‐hybridization with internal, transcript‐specific oligonucleotides. Total RNA from 12 brain and 63 gynecological tumors were then tested for expression of CEA‐related transcripts. None were found in tumors located in the brain, including various mesenchymal and neuro‐epithelial tumors. CEA and NCA transcripts were, however, present in an adenocarcinoma located in the nasal sinuses. In ovarian mucinous adenocarcinomas, we always found co‐expression of CEA and NCA transcripts, and occasionally BGP mRNA. CEA‐related transcripts were also found in some serous, endometrioid and clear‐cell ovarian carcinomas. CEA, NCA and BGP transcripts were present in endometrial carcinomas of the uterus and cervical carcinomas, whereas uterine leiomyomas were completely negative. No transcripts were found from CGM 1, CGM6, CGM7 or from PSG genes in any of the tumors tested. The PCR data were compared with immunohistochemical investigations of ovarian tumors at the protein level using CEA (26/3/13)‐, NCA‐50/90 (9A6FR) and NCA‐95 (80H3)‐specific monoclonal antibodies.
Immunogenic properties of retroviral protein p15e from drug‐treated murine mastocytoma P815
Authors: Grohmann U. , Fioretti M. C. , Binaglia L. , Belladonna M. L. , Bianchi R. , Puccetti P.
Published: February 2007

A triazene‐xenogenized tumor sub‐line was derived from the mouse mastocytoma cell line P815 following several transplant generations in vivo on DTIC. The highly immunogenic P815/ DTIC variant line expressed new CTL‐defined antigens. Novel antigens were also detected by antibodies in immunoprecipitation and by Western blot analysis. Upon immunoprecipitation of metabolically labeled cells, one such variant‐specific 20‐kDa antigen was shown to be related to retroviral envelope protein p15E. When injected intrasplenically into recipient mice, the electroblotted nitrocellulose‐bound 20‐kDa antigen resulted in increased frequency in CTL precursors to P815/DTIC cells. In addition to previous data in the L5I78Y/DTIC tumor‐model system, these data suggest that expression of aberrant, retrovi‐rus‐related proteins may be a common finding in different parental tumors xenogenized by triazene treatment.
The association between epstein‐barr virus and chinese hodgkin's disease
Authors: Zhou Xiao‐Ge , Hamilton‐Dutoit Stephen J. , Pallesen Gorm , Yan Qin‐Han
Published: February 2007

Epstein‐Barr virus (EBV) can be detected in Hodgkin and Reed‐Sternberg (HRS) cells in about one‐half of cases of Hodgkin's disease (HD) in Western countries. To determine whether EBV is also associated with HD in a developing country such as China, we studied paraffin sections from 28 Chinese cases of HD for expression of latent membrane protein‐1 (LMP‐1) and EBV‐encoded small RNA (EBER‐I), using immuno‐histology and RNA/RNA in situ hybridization respectively. The cases were selected from a large series of Chinese lymphomas following histological and immunophenotypical revision. EBV gene expression was found in HRS cells in 17/28 cases, and was related to histological sub‐type, being present in 10/11 of mixed cellularity, 6/14 nodular sclerosis, 0/1 lymphocytic predominance, 0/1 lymphocytic depletion, and 1/1 unclassified HD. The 2 methods for detecting EBV gene expression gave similar results, except in one case of nodular sclerosis, in which HRS cells were negative for EBER‐1, but weakly positive for LMP‐1. In 5/12 cases with EBER‐negative HRS cells, rare small or medium‐sized lymphocytes expressed EBER‐1 but not LMP‐1. These results suggest that (i) Chinese HD is frequently associated with EBV; (ii) the proportional frequency and sub‐type distribution of EBV‐positive HD are similar in China and in the West; (iii) both LMP‐1 immunohistology and EBER in situ hybridization reliably detect EBV in HRS cells in routine biopsies, but the former is simpler and less resource‐consuming to perform.
Tumor angiogenesis is an independent prognostic indicator in primary breast carcinoma
Authors: Toi Masakazu , Tominaga Takeshi , Kashitani Jitsuo
Published: February 2007

Clinical importance of tumor angiogenesis, especially its significance as a prognostic indicator, was examined in 125 primary breast‐cancer patients. The grade of neovascularization was assessed by the vessel density which was obtained by immunocytochemical staining for factor VIII antigen. Postoperative survey demonstrated that the vessel density is a statistically significant predictor of relapse‐free survival (median follow‐up period: 62 months). Patients with over 100 counts of factor‐VIII antigen‐positive cells per 200 x field in the most active areas of neovascularization showed significantly poorer prognosis than those with less than 100 counts. The prognostic value of the vessel density was also confirmed by another evaluation method using immunocytochemical staining to CD‐31 which is a platelet/endothelial cell adhesion molecule. A significant difference in relapse‐free survival rate was shown between patients having higher counts of CD‐31 positive cells and those having lower counts. Breakdown analysis stratified by nodal status showed that the vessel density was a significant prognostic indicator in node‐negative and node‐positive patients. Multivariate analysis indicated that the vessel density is an independent prognostic indicator in primary breast‐cancer patients.
Human papillomavirus in vulvar squamous‐cell carcinoma and in normal vulvar tissues: A search for a possible impact of HPV on vulvar cancer prognosis
Authors: Hørding Ulla , Bock Johannes E. , Kringsholm Birgitte , Andreasson Benny , Visfeldt Jakob , Daugaard Søren
Published: February 2007

Paraffin‐embedded sections of vulvar squamous‐cell carcinomas and of normal vulvar tissues were examined for HPV types 6, 11, 16, 18 and 33 by the polymerase chain reaction. Overall, 19 of 62 tumours harboured HPV DNA of types 16, 18 or 33. HPV types 6 and 11 were not detected. HPV DNA was found in 61% of tumours with adjacent intraepithelial neoplasia (VIN III), and in 13% of tumours without associated VIN III. HPV DNA was not detected in any of 101 normal vulvar tissues. HPV DNA was found more often in younger women, in patients with VIN III‐associated tumours, and in those with multicentric anogenital neoplasia. This points to the existence of a subset of vulvar carcinomas preceded by intraepithelial neoplasia, with HPV as a major factor in carcinogenesis. HPV also seems to be an important factor in the development of multiprimaries in these patients. The 2 groups of patients with vulvar carcinoma did not differ with regard to prognosis, as estimated by the risk of recurrence after primary surgery.
Hair dyes, analgesics, tranquilizers and perineal talc application as risk factors for ovarian cancer
Authors: Tzonou Anastasia , Polychronopoulou Argy , Hsieh Chung‐Cheng , Trichopoulos Dimitrios , Rebelakos Apostolos , Karakatsani Anna
Published: February 2007

In a hospital‐based case‐control study of ovarian cancer conducted in Athens (1989‐1991), 189 women with histologically confirmed common malignant epithelial tumors of the ovary were compared with 200 hospital visitor controls. All interviews were conducted by personal interview in the 2 participating hospitals and the data were analyzed by modelling through logistic regression, controlling for demographic and reproductive variables. Tranquilizing and hypnotic drugs (mostly diazepam) were not associated with risk of ovarian cancer: the adjusted relative risk (RR) and 95% confidence interval (CI) were 0.96 (0.57 to 1.63), whereas use of analgesics (mostly salicylates) was associated with significantly reduced risk of the disease (RR 0.51; CI 0.26 to 1.02). There was no evidence that perineal application of talc was associated with increased risk (RR 1.05; CI 0.28 to 3.98) but the frequency of reporting talc use was low in the study population. There was a statistically significant (p for trend 0.007) and a dose‐dependent association between hair dyeing and risk of ovarian cancer. Compared to never‐users, women dyeing their hair up to 4 times per year had a relative risk of 1.74 (0.91 to 3.32) whereas those dyeing their hair 5 or more times per year had a relative risk of 2.16 (1. 19 to 3.89).
Incidence of infiltrating cancer following superficial bladder carcinoma
Authors: Levi Fabio , Vecchia Carlo La , Randimbison Lalao , Franceschi Silvia
Published: February 2007

Patients with histologically confirmed first diagnosis of superficial bladder carcinoma notified to the population‐based cancer registry of the Swiss Canton of Vaud during the calendar period 1974‐90 were actively followed‐up to December 31, 1990 for the occurrence of a subsequent invasive tumour of the urinary bladder. Among 1,012 incident cases of superficial bladder neoplasms, followed for a total of 6,286 person/years at risk, 93 infiltrating tumours of the urinary bladder were diagnosed. Only 5.3 cases were expected on the basis of the general population of the canton. The overall standardized incidence ratio (SIR) was 17.5 (95% confidence interval, CI: 14.2–21.7). The SIR was significantly greater for females than for males. The SIR was highest between I and 4 years following registration of noninfiltrating cancer, and declined thereafter. The cumulative risk of invasive bladder cancer was 7%, 13%, and 16%, after 5, 10 and 15 years, respectively. This work provides population‐based, accurate and reliable estimates of the risk of invasive bladder cancer following non‐infiltrating cancers. Although the overall relative risk was almost 20‐fold higher than in the general population, the cumulative risk of developing an invasive bladder cancer was only 16% at 15 years.
Defective vasculature in fibronectin‐receptor‐deficient cho cell tumors in nude mice
Authors: Schreiner C. L. , Fisher M. , Bauer J. , Juliano R. L.
Published: February 2007

The level of expression of the α5/β1 integrin fibronectin receptor(FnR) strongly affects the growth rate of CHO cell tumors in nude mice. Here we report that α5/β1 expression also influences the organization of the tumor vasculature. Tumors formed from CHO clones defective in FnR expression have a leaky vasculature that gives them a hemorrhagic appearance. Tumors from wild‐type CHO cells, or from FnR‐deficient CHO clones transfected with constructs coding for the α5 integrin subunit, have an intact, non‐leaky vasculature. In tumors from FnR‐deficient cells, the endothelial lining of blood vessels is sparse, and red cells and plasma proteins can be detected in the tumor parenchyma. In tumors from cells expressing the α5/β1 FnR, tumor vessels are circumscribed by a lining of von‐Willebrand‐factor‐positive endothelial cells, and blood cells and proteins are confined to the vessel lumina. Thus, the level of expression of the α5/β1 FnR in the tumor parenchymal cells can influence the development of tumor vasculature. Since α5/β1 is vital to the organization of the extracellular matrix, one possibility is that altered matrix assembly contributes to the defective vascularization seen in α5/β1 ‐deficient tumors.
Association of insulin‐like growth‐factor‐I‐induced DNA synthesis with phosphorylation and nuclear exclusion of p53 in human breast cancer MCF‐7 cells
Authors: Takahashi Kazuhide , Suzuki Katsuo
Published: February 2007

Human breast cancer MCF‐7 cells, growth‐arrested by serum starvation, were stimulated with recombinant human insulinlike growth factor‐1 (IGF‐I). An increase in DNA synthesis was induced 20 hr later, which was as effective as that induced by serum. The increase in DNA synthesis was significantly inhibited either by antibody to the IGF‐I receptor or by the tyrosine kinase inhibitor, methyl‐2,5‐dihydroxycinnamate (2,5‐MeC). The IGF‐l‐induced DNA synthesis coincided with an elevated level of phosphorylation of p53 on tyrosine and an alteration in the subcellular distribution of the protein from the nucleus to the cytoplasm. Whereas the increases in DNA synthesis and p53 phosphorylation were inhibited by antibody to the IGF‐I receptor and by 2,5‐Mec, the nuclear exclusion of p53 was prevented by the antibody and also, although not significantly, by 2,5‐Mec. The results suggest that growth stimulation of MCF‐7 cells by IGF‐I is accompanied by tyrosine phosphorylation and nuclear exclusion of p53.
The value of histoquantitative measurements in prognostic assessment of renal adenocarcinoma
Authors: Eskelinen Matti , Aittooja Leena , Hall Outi , Lipponen Pertti , Syrjänen Kari
Published: February 2007

In a series of 135 patients with renal‐cell carcinomas (followed up for a mean of 9.5 years), a variety of clinical and histological factors were analyzed in relation to morphometric measurements of the nuclear parameters in the primary tumours to establish their value as prognostic factors. Clinical, histological and morphometric factors were significantly interrelated in that the metastatic high‐grade tumours had larger nuclei, larger variation in nuclear size and shape, and were also rapidly proliferating. In a univariate analysis, the most important clinical predictor of recurrence‐free survival (RFS) was T category, followed by combined nuclear grade, N category, nuclear grade, tumour size, sex and M category. The most important quantitative predictor of RFS was the mean area of the 10 largest nuclei (NAIO), the mean of the longest nuclear axis (Dmax), SD of nuclear area (SDNA), the volume‐corrected mitotic index (M/V index), inflammatory‐cell reaction, SD of nuclear perimetry (SDPE), and the mean of nuclear area (NA). M category, T category, combined nuclear grade, nuclear grade and N category were significantly related to patient survival. Of the quantitative variables, M/V index, Dmaxand NA were significant predictors of survival in a univariate analysis. Females had longer RFS than men, and density of tumour‐infiltrating lymphocytes (TIL) referred to an increased risk of recurrent tumour in both sexes. In a multivariate analysis, the RFS was independently predicted by the clinical stage, female sex and mitotic frequency/mm2, while nuclear parameters or nuclear grading had no independent prognostic value. The extent of the primary tumour was the single most important determinant of survival, followed by the proliferation rate of the tumour. In local TI‐2NOMO tumours, mitotic frequency/mm2was the only independent prognostic factor for RFS. The clinical stage, mitotic frequency/mm2, nuclear grade and density of TIL were independent predictors in Cox's analysis. In these local tumours, mitotic frequency/mm2of neoplastic epithelium was the only independent prognostic factor. The results indicate that although an accurate prognostic evaluation of renal‐cell carcinomas can be based on subjective nuclear grading and histoquanti‐tative measurements of nuclear parameters, the simple assessment of mitotic frequency alone supplies most of the prognostic data, particularly in local tumours.
Cutaneous malignant melanoma: Association with height, weight and body‐surface area. A prospective study in Norway
Authors: Thune Inger , Olsen Anne , Albrektsen Grethe , Tretli Steinar
Published: February 2007

To investigate whether anthropometric indices as body height, body‐mass index (BMI) and body‐surface area (BSA) have any influence on the risk of cutaneous malignant melanoma (CMM), we analyzed height and weight data from a Norwegian screening survey running between 1963 and 1975 and comprising 1.3 million individuals followed for 14 to 26 years. Among females, CMM was diagnosed in 2814 incident cases, while the number among males was 2144 during 22,988,345 person‐years of follow‐up. With adjustment for age, birth cohort, geographic region and BMI, the risk for CMM increased significantly with increasing quintiles of height in both sexes. The tallest persons were at greater relative risk than the shortest ones: 1.60 and 1.59 in males and females respectively. This positive association was also found for CMM localized on face and trunk in both sexes, and in the lower limbs only in females. Males in the highest quintile of BMI had a relative risk for CMM of 1.26 compared with the lowest quintile, while a non‐significant negative association were found in obese females. This negative association increased and was significant for CMM located on the trunk and lower limbs in females. Among males we observed a consistent association between BSA and risk of CMM similar to that for height. These relationships between anthropometric indices and CMM are discussed according to factors which promote growth, among which nutrition, social class and hormones are the most important.
The value of hepatitis B x antigen as a prognostic marker in the development of hepatocellular carcinoma
Authors: Zhu Minghua , London W. Thomas , Duan Ling‐Xun , Feitelson Mark A.
Published: February 2007

Chronic infection with hepatitis‐B virus (HBV) is associated with high risk for the development of hepatocellular carcinoma (HCC). Several studies have implicated that the X gene product(s) of HBV are important to the pathogenesis of HCC. This study tests the hypothesis that immunohistochemical detection of hepatitis B x antigen (HBxAg) is closely associated with HCC. The patterns of HBxAg were determined by staining in tumor and non‐tumor liver sections from 30 Chinese patients with HBV‐associated HCC, and the results were compared with other markers of infection. HBxAg was the most prevalent marker of HBV infection both in tumor and in non‐tumor tissues of HCC patients, as compared with the hepatitis‐B surface and core antigens. This pattern was observed among carriers as well as several patients who were HBsAG in serum. The HBxAg staining results were validated by Southern blotting with an X‐region probe and by Western blotting with anti‐HBx. These results suggest that the persistence of HBxAg is important to the pathogenesis of early HCC and that HBxAg expression in the liver during chronic HBV infection may be an important prognostic marker for the development of HCC.
Expression of epstein‐barr‐virus‐related nuclear antigens and B‐cell markers in lymphomas of SIV‐immunosuppressed monkeys
Authors: Li Su Ling , Feichtinger H. , Kaaya E. , Biberfeld P. , Migliorini P. , Putkonen P. , Biberfeld G. , Middeldorp J. M. , Ernberg I.
Published: February 2007

Simian‐immunodeficiency‐virus(SIV)‐infected cynomolgus monkeys develop B‐cell lymphomas in approximately one third of the cases. We have now studied the expression of cynomolgus‐Epstein‐Barr‐virus(cyno‐EBV) nuclear antigens in 13 cyno‐EBV‐carrying SIVsm‐associated monkey lymphomas and established cell lines from 3 of these tumors. Immunoblots of cell lysates were probed with polyspecific and monospecific reagents directed against human EB‐virus EBNA1‐6, and against the membrane protein LMP1. An EBNA2‐cross‐reacting protein was demonstrated in 8 lymphoma tissues (8/13) and in the 3 cell lines derived from the tumors. All tumors expressed a polypeptide with 50 to 55 kDa molecular weight, which cross‐reacted with some antibodies to EBNA1. Absorption experiments with normal monkey tissue showed that this polypeptide was specific for the cyno‐EBV‐carrying lymphoma cells. Equivalents of EBNA3‐6 and LMP1 could not be detected. Immunophenotypical characterization showed that the monkey lymphomas were similar to human HIV‐associated B‐cell lymphomas. Malignant B‐cell lymphomas in experimentally SIVsm‐infected cynomolgus monkeys can be a model for EBV‐associated lymphomagenesis in immunodeficiency states.

This study was supported by grants from the Swedish Medical Research Council, the Swedish Cancer Society and Erik and Edith Fernström's Foundation for Medical Research. We thank Dr. J. Dillner (Department of Virology, Karolinska Institute) for helpful suggestions, Ms. A. de Santiago de Aguilar, Ms. M. Ekman and Ms. R. Gendelman for skillful technical assistance, and Drs. A. Rickinson and M. Rowe, CRC laboratories, for PE‐2 MAb. H. F. was partly supported as a visiting scientist by the European Community Concerted Action “Pathophysiology and immunology of HIV‐related diseases”, and E.K. (visiting scientist from Muhimbili Medical Centre, Dar‐es‐Salaam, Tanzania) was supported by the Swedish Agency for Research Cooperation with Developing Countries (SAREC) and World Laboratory (Project MDC 2).
Induction and release of manganese superoxide dismutase from mitochondria of human umbilical vein endothelial cells by tumor necrosis factor‐α and interleukin‐1α
Authors: Nakata Tsoshiyuki , Suzuki Keiichiro , Fujii Junichi , Taniguchi Naoyuki , Ishikawa Mutsuo
Published: February 2007

The effects of tumor necrosis factor‐α (TNFα) on cultured human umbilical vein endothelial cells (EC) and 2 cancer cell lines, AS49 and ME 180, were compared. The effects of interleukin‐ 1α (IL‐ 1α) on EC were also examined. While A549 cells were fairly resistant to the cytolytic effects of TNFα and IL‐1α, ME 180 cells were sensitive. EC were also less sensitive to TNFα than ME 180 cells, as judged by viability of individual cells and by the release of lactate dehydrogenase (LDH) into the medium. Both manganese superoxide dismutase (Mn‐SOD) and its mRNA were markedly induced by these cytokines in EC and in A549 cells but not in ME 180 cells. The levels of Mn‐SOD in the conditioned medium of EC were markedly increased after stimulation with cytokines, whereas those in ME180 and A549 cells were relatively low. The amount of Mn‐SOD released appears to be comparable to that from cells lysed due to the cytocidal effect of cytokines, as assessed by measuring intra‐ and extra‐cellular LDH activity. These data suggest that, in vivo, the TNFα and IL‐1α produced by cancer cells and other cells may induce Mn‐SOD in vascular endothelial cells as well as other host tissues, resulting in release of a relatively large amount of this protein into the serum.
Parity and hepatocellular carcinoma. A population‐based study in sweden
Authors: Lambe Mats , Trichopoulos Dimitrios , Hsieh Chung‐Cheng , Ekbom Anders , Pavia Maria
Published: February 2007

The association between parity and hepatocellular carcinoma (HCC) was studied using a data‐base generated by linking 2 Swedish nation‐wide registries; the Cancer Registry and the Fertility Registry. Among women born between 1925 and 1960, 133 patients with HCC recorded in the Cancer Registry between 1958 and 1984 were compared with 665 age‐matched controls. In this nested case‐control study there was no positive association between parity, age at first birth or frequency of twinning on the one hand, and risk of HCC on the other. It appears that the positive association between parity and HCC previously reported is limited to cases of HCC caused by chronic infection with hepatitis B virus; these cases represent only a small fraction of HCC cases in Sweden.
Epstein‐barr virus subtype distribution in angioimmunoblastic lymphadenopathy
Authors: Borisch Bettina , Caioni Massimo , Hurwitz Nina , Dommann‐Scherrer Corina , Odermatt B. , Waelti Ernst , Hubert R. Hubert , Kraft Rainer , Laissue Jean
Published: February 2007

The tissues of 16 patients bearing a T‐cell lymphoma of angioimmunoblastic lymphadenopathy type (AILD‐TCL) were investigated for the distribution of Epstein‐Barr virus (EBV) subtypes 1and 2. EBV‐association had been proven in these cases by polymerase chain reaction (PCR) for EBV‐DNA, in situ hybridization (ISH) for EBV‐encoded small nuclear RNAs (EBER) and immunohistology for EBV‐encoded latent membrane protein (LMP). PCR and EBER‐ISH produced mostly identical results, but some cases were positive with only one of the 2 methods employed. LMP was detected in a few large cells of 8/13 cases. Twelve cases were investigated for the distribution of EBV subtypes. One case contained EBV genome of subtype 2, 3 cases contained subtype 1 and 4 cases contained both subtypes. Four cases could not be typed. These finclings suggest that in AILD, as in AIDS‐associated lymphomas and lymphomas of the lethal midline granuloma type, subtype 2 of EBV may occur, perhaps in relation to an immunodysfunction developing progressively in these patients.
Immune recognition of human colonic‐tumour‐associated muc‐2 mucins using an anti‐peptide antibody
Authors: Price Michael R. , Sekowski Michael , Ladányi Andrea , Uray Katalin , Ma Yuejie , Durrani Lindy , Tendler Saul J. B.
Published: February 2007

In human intestinal malignancy, alterations occur in the expression of mucins defined by the MUC‐2 gene. These changes include the unmasking of epitopes in the mucin protein core. In order to probe these modifications associated with mucins of the malignant phenotype, a monoclonal antibody (MAb) was developed against synthetic peptide with a sequence based upon that of the protein core of the MUC‐2 mucin. The antibody (designated 996) was shown to recognize a high‐molecular‐weight glycoprotein from colonic carcinoma tissue. The material reacted uniformly with Concanavalin A but variably with other lectins, indicating heterogeneity in the associated oligosaccharide side chains. The protein core was accessible both to 996 antibody binding and to degradation with proteases. Immunization with the affinity‐purified mucin‐like material elicited antibodies reactive with both the immunogen and the synthetic peptides, confirming the immunogenic character of protein‐core determinants. Epitope mapping studies, using synthetic peptides in solution and synthetic peptides tethered to the heads of plastic pins, indicated that the minimum epitope for the 996 antibody is a tetramer of T G T Q. Antibody interaction with the glutamine (Q) residue was determined to be of major importance in the antigen‐antibody reaction. The findings illustrate the characterization of an anti‐peptide antibody which may be used to probe alterations in MUC‐2 mucin expression associated with human intestinal malignant disease.
An analysis of a melanoma epidemic
Authors: Burton Robert C. , Coates Marylon S. , Hersey Peter , Roberts Graeme , Chetty M. P. , Chen Stan , Hayes Monica H. , Howe Chris G. , Armstrong Bruce K.
Published: February 2007

Between 1986 and 1988 the annual incidence of invasive melanoma in the Hunter area of New South Wales, Australia, almost doubled to 52.5 per 100,000 in men and 42.9 per 100,000 in women. These rates have been maintained and are similar to those reported for 1987 in Queensland, Australia, which are the highest in the world. Most of the increase in incidence was in melanomas less than 1,50 mm in thickness, and adults of both sexes and all ages were affected. Thicker melanomas also increased in incidence but only in adults 45 years and older, and mainly in men. An analysis of health insurance data on treatment of skin lesions and data from a histopathology laboratory suggested that diagnosis and treatment of skin lesions generally in the Hunter area had increased almost 2‐fold over this period. Advancement of the time of diagnosis and a real increase in incidence were likely explanations for some of the observed trends. Increasing diagnosis of a non‐metastasising form of thin melanoma, consequent upon increasing removal of pigmented skin lesions by medical practitioners, may also explain some of the observed increase in the incidence of the disease. This possibility has important implications for proposed population screening programs, and methods are needed to distinguish such lesions, if they exist, from potentially fatal melanoma.
Stromelysin‐3 expression in breast cancer biopsies: Clinico‐pathological correlations
Authors: Hähnel Erika , Harvey Jennet M. , Joyce Raymond , Robbins Peter D. , Sterrett Gregory F. , Hähnel Roland
Published: February 2007

Stromelysin‐3 expression was studied by Northern blotting in 222 tissue samples including primary and metastatic breast carcinoma and normal breast tissue. Uninvolved breast tissue from mastectomy specimens, normal breast tissue from reduction mammoplasties and normal lymph nodes did not contain stromelysin‐3 mRNA. About 62% of primary and metastatic breast carcinomas, but only I of 10 in situ ductal carcinomas, expressed stromelysin‐3. Stromelysin‐3 mRNA was found more often in estrogen‐receptor‐positive carcinomas and in histological grade‐l carcinomas. There was no significant correlation between stromelysin‐3 expression and other prognostic factors, including tumor size, lymph‐node involvement, age of patient, vascular invasion and cathepsin‐D.
Serologic response in human papillomavirus‐associated invasive cervical cancer
Authors: Viscidi Raphael P. , Sun Yeping , Tsuzaki Brian , Bosch F. Xavier , Munoz Nubia , Shah Keerti V.
Published: February 2007

Human papillomavirus (HPV) transforming proteins E6 and E7 are uniformly expressed in HPV‐associated cervical cancer. Our objective was to measure antibodies to HPV‐16 E6 and E7 proteins in cervical cancer patients using an assay which would detect antibodies to conformational epitopes. Serum specimens obtained from two case‐control studies of HPVs and cervical cancer were tested. The studies were performed in Cali, Colombia, South America and in 9 provinces of Spain. Cases consisted of women with invasive cervical cancer associated with HPV‐16 or other HPV types and women with HPV‐16‐associated high‐grade cervical intra‐epithelial neoplasia (CIN‐3). Controls for invasive cases and CIN‐3 cases were women who had no cytologic abnormalities and who were matched for age and country of residence. Serum antibodies to HPV‐16 E6 and E7 proteins were detected by radio‐immunoprecipitation of in vitro translated proteins. Antibodies to the E6 and E7 protein were observed among 56% and 43%, respectively, of invasive cases and 1.7% and 4.1%, respectively, of controls. Antibodies to either protein were detected in 72% of sera from invasive cases and 5.8% of sera from controls. High antibody reactivity and antibodies to both proteins were found almost exclusively in invasive cases. The frequency of antibodies to the E6 protein and the E7 protein among CIN‐3 cases did not differ significantly from the CIN‐3 controls. Five women with HPV‐18‐associated invasive cervical cancer were negative for serum antibody to HPV‐16 E6 and E7 proteins. Antibodies to HPV‐16 E6 and E7 proteins appear to be partially virus‐specific and disease state‐specific markers of HPV‐associated cervical cancer.
The predominant mRNA class in HPV16‐infected genital neoplasias does not encode the e6 or the E7 protein
Authors: Bohm S. , Wilczynski S. P. , Pfister H. , Iftner T.
Published: February 2007

Human papillomavirus (HPV) type 16 is strongly implicated in the development of progressive neoplasias of the uterine cervix. Its oncogenic potential is decisively determined by the activity of the early gene products E6 and E7. To look for changes in the expression of these genes during tumour progression we cloned subgenomic fragments of HPV 16 into RNA expression vectors, which allowed the generation of 35S‐labelled riboprobes specific for distinct mRNA classes. Four constructs were made to differentiate between transcripts starting upstream of the E6 ORF or the E1 ORF, and one probe was specific for unspliced E6/E7 region transcripts. Five other constructs were used to identify transcripts covering the E1, E2, E4, L1 and L2 regions. With the help of these constructs, we analyzed by in situ hybridization 2 low‐grade intraepithelial neoplasias of the vulva, I high‐grade neoplasia of the cervix as well as 4 vulvar and 3 cervical carcinomas. Transcripts from the E1, E2, E4, L1 and L2 region that were consistently detected in the differentiated layers of benign lesions were variably expressed in precancers and carcinomas. None of the investigated cases revealed detectable amounts of unspliced E6/E7 transcripts with a coding potential for a full‐length E6 protein. In benign lesions, the E7 transcripts were confined to isolated nuclei of differentiated cells, whereas high‐grade lesions and invasive cancers showed elevated levels of equally distributed E7‐specific signals in the cytoplasm of all tumour cells. The most abundant transcripts observed in intraepithelial neoplasias and in invasive cancers appear to initiate within ORF E7 and therefore have no coding potential for full‐length E6 and E7 proteins. Our data show that the actual level of E7‐specific transcripts in cancers is lower than anticipated from earlier studies using an ORF E6/E7‐specific probe that hybridizes with the 5′‐ends of the abundant mRNA class.
Enhanced expression of LFA‐3 on human T‐cell lines and leukemic cells carrying human T‐cell‐leukemia virus type 1
Authors: Imai Toshio , Tanaka Yoshikazu , Fukudome Kenji , Takagi Shin , Araki Koichi , Yoshie Osamu
Published: February 2007

Previously, we studied the surface expression of LFA‐1 (CD 11 a/CD 18) and ICAM‐1 (CD54) in a panel of 16 human T‐cell lines and found that those carrying HTLV‐1 pro‐viruses expressed high levels of ICAM‐1. Enhanced expression of ICAM‐1 was also seen in fresh leukemic cells from ATL patients. In the present study, we systematically examined the surface expression of 19 different cell‐adhesion molecules (CAMs) (the immunoglobulin superfamily, the homing receptors, the integrins, etc.) in a panel of 20 human T‐cell lines (6 HTLV‐1‐negative lines and 14 HTLV‐1‐positive lines). The surface expression of LFA‐3 (CD58) was constantly enhanced in HTLV‐1‐positive T‐cell lines except for MT‐1, an ATL‐derived cell line, which was devoid of a number of CAMs including ICAM‐1. LFA‐3 proteins and mRNA were found at strongly increased levels in HTLV‐1 ‐positive T‐cell lines.
Uncoordinate induction and differential regulation of hla class‐I and class‐II expression by γ‐interferon in differentiating human neuroblastoma cells
Authors: Ponzoni Mirco , Guarnaccia Francesco , Corrias Mariavaleria , Cornaglia‐Ferraris Paolo
Published: February 2007

Recombinant γ‐interferon (IFN‐γ) has recently been shown to be one of the most effective inducers of neuroblastoma (NB) cell differentiation. Since increasing evidence indicates that expression of MHC class‐I and class‐II antigens by tumour cells is important for immunorecognition and cell targeting, we tested whether induction of NB cell differentiation by IFN‐γ is followed by expression of HLA class‐I and class‐II molecules. LAN‐5 human NB cell line completely lacks HLA class‐I antigens. Their expression was induced in a dose‐dependent manner by IFN‐γ. HLA class‐II molecules are also absent on LAN‐5 cells, but only DP antigens were dose‐dependently induced by IFN‐γ, while DR and DQ molecules were unaffected by the treatment. To confirm and extend the immunological data to all the class‐II molecules, we performed Northern blot analysis, observing that DPα mRNA was induced in a dose‐ and time‐dependent manner. DOβ and DZα genes were also induced peaking after 3 days of IFN‐γ treatment. DRβ and DQβ genes, which were not induced by IFN‐γ, gave a normal pattern of enzyme restriction by Southern blot. To get an insight into the regulation of HLA class‐II gene expression in the neuronal model, we measured the decline of the steady‐state HLA class‐II mRNA. DOβ mRNA rapidly returned to baseline level after removing IFN‐γ, while the decay rates of DPα and DZα mRNA were very slow. This might indicate different regulation at the post‐transcriptional level for DOβ mRNA with respect to DPα and DZα mRNA. To strengthen these findings we evaluated the half‐lives of the mRNA after IFN‐γ induction by means of actinomycin D treatment. HLA‐DOβ mRNA had a shorter half‐life, while DZα and DPα had a longer decay rate. Finally, we report that treatment of LAN‐5 cells with cycloheximide did not alter the rate of transcription of the HLA‐DPα gene, suggesting that no protein factor(s) is/are needed to maintain DPα gene expression.
Decreasing the level of translation initiation factor 4E with antisense rna causes reversal of ras‐mediated transformation and tumorigenesis of cloned rat embryo fibroblasts
Authors: Rinker‐Schaeffer Carrie W. , Graff Jeremy R. , De Benedetti Arrigo , Zimmer Stephen G. , Rhoads Robert E.
Published: February 2007

Transformation of cloned rat embryo fibroblasts (CREF) with the T24‐ras oncogene results in loss of contact inhibition, growth in soft agar and tumor formation in nude mice. Previously we showed that in such cells (CREF T24), the phosphorylation rate of protein synthesis initiation factor 4E (elF‐4E) is increased, correlating with an increase in the general rate of protein synthesis. In the present study, we have expressed antisense RNA complementary to elF‐4E mRNA in CREF T24 cells using a stably integrated vector. Cells expressing antisense RNA (CREF T24/AS) contained 30‐50% of the normal level of elF‐4E and exhibited many of the properties of untransformed cells. CREF T24 had a spindle‐shaped, refractile appearance, whereas CREF T24/AS grew in ordered, parallel patterns and exhibited contact inhibition similar to untransformed CREF. The rates of growth and protein synthesis in CREF T24/AS were decreased compared to CREF T24 but were not as low as in CREF. The efficiency of growth in soft agar was 11‐fold lower for CREF T24/AS compared with CREF T24. The latency period for tumor formation in nude mice was increased from 8 days for CREF T24 to 17‐27 days for CREF T24/AS and various clonal lines derived from them. Cell lines established from these CREF T24/AS‐derived tumors were shown to have partially regained the elF‐4E levels characteristic of CREF T24. These results demonstrate that many of the phenotypic alterations associated with ras‐induced malignant transformation can be reversed by a moderate reduction of the translational initiation capacity and therefore may be mediated through a translational mechanism.
Circumvention of tamoxifen resistance by the pure anti‐estrogen ICI 182, 780
Authors: Hu X. F. , Veroni M. , de Luise M. , Wakeling A. , Sutherland R. , Watts C. K. W. , Zalcberg J. R.
Published: February 2007

Both primary and acquired resistance to the growth‐inhibitory effects of anti‐estrogens (e.g., tamoxifen) limits the clinical usefulness of these drugs in the treatment of breast cancer. The new, steroidal anti‐estrogen ICI 182,780 was tested for its ability to inhibit the proliferation of a tamoxifen‐resistant variant of the parental MCF‐7 human breast‐cancer cell line. Two cell lines cloned from the MCF‐7 line were used for these experiments: a tamoxifen‐sensitive line, MCF 5‐21, and a tamoxifen‐resistant line, MCF 5‐23. Compared with tamoxifen, ICI 182,780 appeared to be 150 and 1540 times more effective in inhibiting cell growth in the 5‐21 and 5‐23 sub‐lines respectively. ICI 182,780 completely circumvented tamoxifen resistance at a concentration of (5 to 10) × 10−9 M in this model. Based on IC50 concentrations, the 5‐23 line was 22‐fold more resistant to tamoxifen than the 5‐21 line, but only 2‐fold more resistant to ICI 182, 780, reducing relative resistance by 10‐fold in the resistant line. There were no differences in ER parameters between the 2 lines. ER numbers/cell were: 40500 and 34800 and the KD 0.48 and 0.15 × 10−9 M in the 5‐21 and 5‐23 cells respectively. In the 5‐23 cells, the concentrations of ICI 182, 780 and tamoxifen resulting in a 50% inhibition of 3 H‐estradiol binding were 2.3 × 10−8 M and 1 × 10−6 M, respectively (cf. estradiol 0.89 × 10−9 M). Thus, one potential mechanism for the increased effectiveness of ICI 182,780 may relate to the increased affinity of this drug for the estrogen receptor as compared with tamoxifen.