bluuebunny/crossref_metadata_2025 huggingface dataset:
Biodistribution of lymphokine‐activated killer (LAK) cells in WAG rats after hepatic‐artery or jugular‐vein infusion
Authors: Kuppen Peter J. K. , Marinelli Andreas , Camps Jan A. J. , Pauwels Ernest K. J. , van de Velde Cornells J. H. , Fleuren Gert Jan , Eggermont Alexander M. M.
Published: February 2007

This study deals with the biodistribution of syngeneic radiolabeled lymphokine‐activated killer (LAK) cells in Wag rats after infusion via the hepatic artery or the jugular vein. The biodistribution of IIIIndium‐labeled LAK cells was evaluated using serial whole‐body gamma camera imaging. Furthermore, we investigated 2 factors that might influence the biodistribution of these effector cells: purity of LAK cells and administration of interleukin‐2 (IL‐2). After injection of IIIIndium‐labeled LAK cells via the hepatic artery or via the jugular vein we could detect important differences in the biodistribution pattern up to 5 hr after injection. LAK cells administered via the jugular vein were all found in the lungs up to 2 hr after injection and then redistributed to the liver and to the spleen. LAK cells administered via the hepatic artery were all found in the liver after injection and redistributed after 2 hr mainly to the spleen. About 8 hr after injection we could no longer detect any differences in the biodistribution pattern according to the route of administration. Biodistribution was followed for up to 72 hr after injection but the pattern showed no change after 8 hr, whichever the route of administration. A purified adherent‐LAK population, a large granular lymphocyte culture with only 6% T cells, showed the same distribution pattern as standard LAK cells (40% T cells). Infusion of 40,000 units of IL‐2 per day, starting 3 days before and continuing after administration of radio‐labeled LAK cells, accelerated the redistribution of these cells by both routes of administration. We conclude that up to 2 hr after local infusion, a high concentration of LAK cells in the first capillary bed can be obtained. Therefore, local administration of LAK cells may be more effective against tumors. © 1992 Wiley‐Liss, Inc.
Derivation of tumorigenic and non‐tumorigenic mouse alveolar type‐II cell lines from fetal type‐II cells after a combined in vivo/in vitro carcinogen treatment
Authors: Oomen L. C. J. M. , Calafat J. , Have‐Opbroek A. A. W. Ten , Egberts J. , Demant P.
Published: February 2007

Alveolar type‐II cells wens isolated from the lungs of fetuses (day 18 of gestation) of the A/WySnAf (A/Sn) mouse strain, which were treated in utero at day 15 with the directly‐acting carcinogen N‐ethyl‐N‐nitrosourea (ENU). The isolated type‐II cells were again treated with ENU during their initial growth in vitro. After a prolonged culture period, 5 cell lines were obtained, which were identified as type‐II cell lines. Differences between cell lines were found with respect to contact‐inhibited growth, cell doubling time and ability to grow in a serum‐free medium. Two out of the 5 cell lines produced highly invasive type‐II cell carcinomas after s.c. injection of 5 × 106 cells into nude nice. Thus, both tumorigenic and non‐tumorigenic mouse alveolar type‐II cell lines were derived after this combined in vivo and in vitro carcinogen treatment of fetal mouse alveolar type‐ll cells. This offers the possibility of studying in vitro the factors thought to influence lung tumorigenesis in vivo. In addition, our findings strongly suggest that alveolar type‐II cells are the progenitor cells of malignant mouse lung tumors. © 1992 Wiley‐Liss, Inc.
Epidermal growth factor reduces HER‐2 protein level in human ovarian carcinoma cells
Authors: Marth Christian , Lang Thomas , Cronauer Marcus V. , Doppler Wolfgang , Zeimet Alain G. , Bachmair Franz , Ullrich Axel , Daxenbichler Günter
Published: February 2007

Over‐expression of the proto‐oncogene HER‐2 (c‐erbB‐2/neu) in ovarian, endometrial, and mammary carcinoma is an indicator of poor prognosis. Interactions between the epidermal growth factor (EGF) receptor and the HER‐2 protein have been described. The aim of this study was to elucidate the effects of EGF on HER‐2 expression. In the human ovarian carcinoma cell lines HTB‐77, OVCAR‐3, 2780, SKOV‐6, SKOV‐8 and 2774, and the human mammary tumor cell line SKBR‐3, total cellular p185HER‐2 was determined by an ELISA, whereas the surface p185HER‐2 was measured with a living‐cell RIA. Stimulation of these cell lines with either EGF (0.1–30 nM) or TGF‐α (0.1–30 nM) led to a significant reduction in p185HER‐2 expression. The effect was more pronounced in cells with normal HER‐2 expression. A reduction of mRNA levels for p185HER‐2 by EGF was observed in OVCAR‐3 cells but not in the over‐expressing lines HTB‐77 and SKBR‐3. Interestingly, the EGF‐induced effect was not always associated with growth stimulation and was not correlated with the number of EGF binding sites detected by a radioligand assay. Our data indicate that EGF treatment results in a down‐regulation of p185HER‐2. © 1992 Wiley‐Liss, Inc.
Lipoprotein(a) and CA125 levels in the plasma of patients with benign and malignant ovarian disease
Authors: Kuesel Annette C. , Kroft Thérèse , Préfontaine Michel , Smith Ian C. P.
Published: February 2007

Elevated plasma levels of apolipoprotein (a) have been reported earlier in cancer patients. In order to investigate the potential of apolipoprotein(a) as an ovarian tumour marker, plasma apolipoprotein(a) and CA125 levels were measured in healthy women and women with benign or malignant pelvic masses. Among women younger than 49 years, 80% of healthy controls and ovarian cancer patients had apolipoprotein(a) levels below 350 U/I. Among women aged 49 years or older, 46% of healthy controls but 73% of ovarian cancer patients and 77% of women with successfully treated ovarian cancer, had low apolipoprotein(a) levels. For both age groups, apolipoprotei‐n(a) is not a suitable marker for ovarian cancer. No correlation was found between apolipoprotein(a), triglyceride or cholesterol in plasma. Healthy women younger than 49 years had significantly higher CA125 levels than women 49 years or older (20 × 14 U/ml vs. 13 × 12 U/ml, p < 0.005). Levels of CA125 above 35 U/ml were found in 12% of the younger and 4% of the older healthy women, 73% of the younger and 61 % of the older patients with untreated or residual tumours, and in 33% of the younger and 31 % of older patients with no evidence of disease, as well as in 58% of women of both age groups with benign pelvic masses. The sensitivity and specificity of CA125 levels for the detection of cancer were 73% and 74% respectively for women younger than 49 years, and 62% and 78% respectively for women 49 years or older. © 1992 Wiley‐Liss, Inc.
Isolation of exfoliated colonic epithelial cells, a novel, non‐invasive approach to the study of cellular markers
Authors: Albaugh George P. , Iyengar Vasantha , Lohani Althaf , Malayeri Mehran , Bala Shukal , Nair P. P.
Published: February 2007

Human stool is a heterogeneous mixture of non‐digestible food residues, bacteria, cells exfoliated from the gastrointestinal mucosa and other secretory products. We have demonstrated that fresh human stools dispersed in a buffered saline solution can be fractionated over Percoll/BSA gradients to yield 9 discrete bands of cells in the density range of p 1.033 to 1. 139 and which could be further purified over Histopaque 1077. Enzyme‐linked immunoassays (ELISA) for colon‐specific antigen (CSA) and cytokeratins (CK) were positive. Western blot analysis showed the presence of 3 cytokeratin bands in the 40‐kDa to 60‐kDa range suggestive of cytokeratins 8, 18, and 19. Fluorescence flow‐cytometric analysis of these cells using antibodies against CSA, CK, the blood‐group antigens, carcinoem‐bryonic antigen (CEA), non‐mucus‐secreting columnar‐epithelium‐specific MAb PRIA3, and to mucus‐secreting colonic‐epithelium‐specific MAb PR5D5 showed varying degrees of reactivity. Expression of the blood‐group phenotype suggests that cells from the proximal half of the colon had survived the transit, since in the adult the expression of this marker is limited to cells from the proximal region of the colon. In this report we demonstrate the feasibility of studying, non‐invasiveLy, cell‐specific markers on exfoliated cells isolated from stools. The evidence strongly suggests that almost all the cells are of colonic origin. © 1992 Wiley‐Liss, Inc.
Glucose depletion enhances the anti‐tumor effect of TNF
Authors: Volland Sabine , Amtmann Eberhard , Sauer Gerhard
Published: February 2007

Treatment of human carcinoma xenotransplants in athymic mice with recombinant human tumor necrosis factor (rh TNF) causes necrosis mainly in the central parts of the tumors, while peripheral sections remain mitotically active. As tumors are known to be supplied with adequate glucose exclusively in their periphery, the influence of the lack of glucose on the cytotoxic activity of rh TNF was studied. The absence of glucose enhanced the killing of tumor cell lines by rh TNF in tissue culture. Meth‐A, a cell line known to be resistant to TNF in vitro but highly sensitive to it in vivo, was readily killed in tissue‐culture medium lacking glucose. All non‐transformed cell lines tested were found to be resistant to rh TNF, regardless of the presence or absence of glucose. In tumor‐bearing mice a reduction of the blood glucose content augmented by insulin led to increased anti‐tumor efficiency of rh TNF. The enhanced anti‐tumor activity was reflected both in histological sections of the tumor xenotransplants, by extensive central necroses, and by reduction of the tumor volumes. © 1992 Wiley‐Liss, Inc.
Induction of protein kinase C down‐regulation by the phorbol ester TPA in a calpain/protein kinase C complex
Authors: Savart Michel , Letard Pascale , Bultel Sandrine , Ducastaing Andre
Published: February 2007

Using a calpain/protein kinase C (PKC) complex, we were able to reproduce, in vitro, the induction of PKC down‐regulation by the phorbol ester 12‐O‐tetradecanoyl‐phorbol‐13‐acetate (TPA) which had been previously observed in cells. We show that TPA initiates this phenomenon by promoting a calpain‐dependent conversion of PKC to the Ca2+ phospholipid‐independent protein kinase M (PKM), at physiological calcium concentrations. This effect of TPA was dependent upon the presence of phosphatidylserine and was observed only when PKC was the substrate for the protease, inactivation of calpain by autolysis not being modified by the presence of TPA. Moreover, PKM generated from the calpain‐PKC complex was resistant to calpain, even after addition of TPA. These results suggest that TPA induces a conformational change in PKC, increasing the affinity of the kinase for calpain and consequently permitting its proteolysis for the basal level of calcium in cells. © 1992 Wiley‐Liss, Inc.
Production of stable phenotypes from 9L rat brain tumor multicellular spheroids treated with 1,3‐bis(2‐chloroethyl)‐1‐nitrosourea
Authors: Barcellos‐Hoff Mary‐Helen , Linfoot Peter A. , Marton Laurence J. , Deen Dennis F.
Published: February 2007

During chemotherapy and regrowth of brain tumors, tumor‐cell heterogeneity, and possibly tumor progression, may change as a result of both the selective forces and mutagenic effects of treatment. We have isolated and characterized drug‐response variants of multicellular rat 9L brain‐tumor spheroids exposed to 1,3‐bis(2‐chloroethyl)‐1‐nitrosourea (BCNU). Ten colonies were isolated from spheroids disaggregated immediately after treatment, and 10 colonies were isolated from treated spheroids disaggregated after 1 week in suspension culture. The sensitivity to BCNU was determined by assays of sister chromatid exchange and colony‐forming efficiency in monolayer cultures of each subline after a 1‐hr exposure to graded doses of BCNU. Three classes of response were found: BCNU sensitivity increased, decreased, or was comparable to that of uncloned, parent 9L cells. Resistant phenotypes were predominant (8/10) in sublines from spheroids disaggregated immediately after treatment, whereas hypersensitive phenotypes (4/8) were isolated only from spheroids disaggregated after 1 week of re‐growth. Since subpopulations isolated immediately after treatment do not have the same biological characteristics as those isolated after a period of regrowth, these data suggest that tumor‐cell heterogeneity may be generated by distinct processes at various times during therapy. The predominance of hypersensitive sublines obtained by the regrowth protocol may have resulted from the recovery of cells that would have died if isolated but were instead able to repair the drug‐induced damage when left in contact with neighboring, possibly resistant cells. Two resistant and two hypersensitive sublines were studied further. © 1992 Wiley‐Liss, Inc.
Influence of the estrous cycle at gamma‐ray exposure on radiation‐induced mammary tumorigenesis in virgin rats
Authors: Inano Hiroshi , Ishii‐Ohba Hiroko , Suzuki Keiko , Yamanouchi Hiroshi
Published: February 2007

Wistar rats received whole body irradiation with 260 cGy gamma‐rays at 10 a.m. of individual phases of their estrous cycle and then had diethylstilbestrol pellets implanted for 1 year. When the radiation was given during di‐estrus II, the highest incidence (73.3%) of mammary tumorigenesis was observed, and mean latency until the first tumor appearance was 8.5 × 0.7 months. Also, rats irradiated on estrus had significantly lower incidence (35.3%) of mammary tumors than those irradiated on pro‐estrus and di‐estrus II, but there was no significant difference in latency period, Iball's indices of total mammary tumors, fibroadenoma plus adenocarcinoma, were 14.3 × 0.9, 28.8 × 2.1, 23.3 × 0.8 and 12.2 × 0.2 in rats irradiated during di‐estrus I, di‐estrus II, pro‐estrus and estrus respectively. The percentage of adenocarcinomas was comparatively uniform (25.0 to 34.8%) throughout the various phases of the estrous cycles. Also, Iball's indices calculated from adenocarcinoma indicated no significant differences between all groups. From our results, the highest incidence of mammary tumors arose in rats after irradiation at di‐estrus II with minimum level of prolactin in serum. We discuss different mechanisms of radiation‐induced and chemical‐carcinogen‐induced tumorigenesis of mammary glands. © 1992 Wiley‐Liss, Inc.
Strong induction of ICAM‐1 in human T cells transformed by human T‐cell‐leukemia virus type 1 and depression of ICAM‐1 or LFA‐1 in adult T‐cell‐leukemia‐derived cell lines
Authors: Fukudome Kenji , Furuse Mikio , Fukuhara Norio , Orita Satoshi , Imai Toshio , Takagi Shin , Nagira Morio , Hinuma Yorio , Yoshie Osamu
Published: February 2007

Sixteen human T‐cell lines were studied for the expression of a cell‐adhesion molecule ICAM‐1 and its counter‐receptor LFA‐1. The cell lines included 3 human T‐cell‐leukemia‐virus‐type‐1 (HTLV‐1)‐negative cell lines derived from acute lymphoblastic leukemia (ALL) and 13 HTLV‐1‐positive cell lines, 7 of them established from cord‐ or peripheral‐blood T cells by in vitro transformation with HTLV‐1, 2 derived from HTLV‐1 carriers, and 4 derived from patients with adult T‐cell leukemia (ATL). In sharp contrast to a basal level of ICAM‐1 in 3 HTLV‐1‐negative ALL cell lines, strong induction of ICAM‐1 was seen in all HTLV‐1‐positive T‐cell lines except for MT‐1, one of the 4 ATL cell lines used in the present study. On the other hand, the expression of LFA‐1 (CD11a and CD18) was more or less similar among the cell lines with and without HTLV‐1. Interestingly, however, 3 out of 4 ATL cell lines (TL‐Oml, H582, HUT102) revealed striking depression of LFA‐1 expression. Several lines of evidence strongly argued against direct involvement of the viral transactivator p40tax or some autocrine cytokines in the induction of ICAM‐1 in HTLV‐1‐positive T‐cell lines. It was also found that ICAM‐1 and LFA‐1 were involved in syncytium formation induced in the co‐culture of HTLV‐1‐positive and HTLV‐1‐negative human T‐cell lines. Implications of constitutive expression of ICAM‐1 for certain clinical manifestations of ATL and of depression of either ICAM‐1 or LFA‐1 during progression of ATL are discussed. © 1992 Wiley‐Liss, Inc.
Bile acids, non‐phorbol‐ester‐type tumor promoters, stimulate the phosphorylation of protein kinase C substrates in human platelets and colon cell line HT29
Authors: Huang X. P. , Fan X. T. , Desjeux J. F. , Castagna M.
Published: February 2007

Protein kinase C (PKC) is the target for a number of tumor promoters. The mechanism underlying the promoting effects of bile acids in colorectal cancer is not understood. We report that sodium deoxycholate (DOC) triggered activation of PKC in physiological conditions. The biphasic effects of DOC upon PKC activation were Ca2+‐stimulated and did not require phosphatidylserine (PtdSer) as phospholipid co‐factor. The optimal rate of activation was obtained at 0.4 mM DOC and reached approximately half the maximal rate of activation obtained in the presence of PtdSer. Similarly to PtdSer, DOC supported diacylglycerol‐as well as phorbol‐ester‐mediated PKC activation. The reciprocal effects of PtdSer and DOC upon PKC in either 0.5 mM CaCl2 or 0.5 mM EGTA suggest that DOC interacts with the phospholipid‐binding domain to elicit PKC activation. DOC‐supported enzyme activation exhibited substrate specificity different from that of PtdSer‐supported enzyme activation. All tested primary and secondary bile acids activated PKC to various extents, with DOC being the most potent. We suggest that amphipathic bile acids acting in a PtdSer‐like manner provide the hydrophobic environment required for PKC activation. Treatment of 32P‐labeled platelets and colonic cells HT29 C1. 19A with DOC enhanced the phosphorylation of endogenous substrates for PKC. Colonic cells responsive at 50 μM DOC, appeared to be 10‐fold more sensitive than platelets. We suggest that direct or indirect activation of PKC by bile acids may account for the promoting effects of these non‐phorbol‐ester‐type tumor promoters. © 1992 Wiley‐Liss, Inc.
Insertion of SMRV‐H viral DNA at the c‐myc gene locus of a BL cell line and presence in established cell lines
Authors: Middleton P. G. , Miller S. , Ross J. A. , Steel C. M. , Guy K.
Published: February 2007

Mutation of the c‐myc gene locus has occurred during in vitro culture of the Burkitt lymphoma cell line Namalwa. Gene cloning and sequencing of this c‐myc gene locus has revealed the insertion of incomplete copies of the Squirrel Monkey Retrovirus‐Human (SMRV‐H) proviral genome. Insertions of the SMRV‐H genome were seen in all Namalwa cell lines tested, with multiple insertions being seen in some sublines. Viral particles with the morphology of type‐D retrovirus, budding from these cells, were seen by transmission EM. New insertions of the genome were produced spontaneously in these cells. The origins of the type‐D retrovirus and its implications for workers in the field are discussed. © 1992 Wiley‐Liss, Inc.
Progressive abrogation of TGF‐β‐1 and EGF growth control is associated with tumour progression in ras‐transfected human keratinocytes
Authors: Game S. M. , Huelsen A. , Patel V. , Donnelly M. , Yeudall W. A. , Stone A. , Fusenig N. E. , Prime S. S.
Published: February 2007

This study examined the response of human keratinocytes in different stages of transformation to exogenous TGF‐β 1 and EGF as well as their receptor and growth‐factor expression. Cells of the spontaneously immortalized HaCaT cell line and c‐Ha‐ras transfected clones (1–6, 1–7, 11–3, 11–4) exhibited different tumorigenic potentials when transplanted to athymic mice. HaCaT‐ and 1–6 cells were non‐tumorigenic, 1–7 cells formed persisting epidermal cysts (benign tumours) and 11‐3 and 11‐4 cells developed into invasive squamous‐cell carcinomas. TGF‐β 1 inhibited thymidine uptake in a dose‐dependent manner, a progressive decrease in response being associated with an increasing malignant potential (HaCaT > 1–6 > 1–7 = 11‐4). HaCaT‐cells and ras‐clones expressed TGF‐β I mRNA at similar levels, but cells of increasing malignant potential secreted markedly less receptor‐binding TGF‐P (HaCaT > 1–6 = 1–7 > 11‐3 > 11‐4) into the culture medium. Whilst ras‐transfected cells expressed fewer TGF‐P receptors than HaCaT cells, there was little difference between TGF‐p receptor number or affinity between the 4 transfected cell clones. The same was true for the TGF‐p receptor types, but Type‐11 receptors were expressed at lower levels by the malignant clones 11‐3 and 11‐4. When HaCaT and ras‐transfected cells were investigated for their response to exogenous EGF, cells were refractory (1–7, 11‐4), partially stimulated (1–6) or fully stimulated (HaCaT). Cells with increasing malignant potential produced increasing amounts of endogenous TGF‐α (11‐4 = 11‐3 > 1–7 = 1–6 > HaCaT). All tumorigenic ras clones expressed higher mRNA levels than HaCaT‐cells. Ras‐transfected clones expressed fewer high‐ and low‐affinity EGF receptors than HaCaT cells with a tendency toward increased numbers of high‐affinity EGF receptors associated with increasing malignant potential (11‐4 = 11‐3 > 1–7 > 1–6) but these changes were associated with a progressive decrease in receptor affinity. The results indicate that tumour progression in human epidermal keratinocytes transfected with c‐Ha‐ras is associated with a progressive abrogation of TGF‐β 1 and EGF growth control. They suggest that the increased autonomous growth potential associated with advanced stages of epithelial tumour progression can be defined more closely using a cellular profile of TGF‐β and EGF. © 1992 Wiley‐Liss, Inc.
TGF‐α production correlates with tumorigenicity in clones of the SW613‐S human colon carcinoma cell line
Authors: Modjtahedi Nazanine , Haddada Hedi , Lamonerie Thomas , Lazar Eliane , Lavialle Christian , Brison Olivier
Published: February 2007

The c‐myc gene is amplified and the c‐Ki‐ras gene is mutated in the SW613‐S human colon carcinoma cell line. Two cell types with different levels of c‐myc amplification are present in the SW613‐S cell population and representative cell clones can be isolated. The clones with a high level of amplification and expression of the c‐myc gene are tumorigenic in nude mice whereas those with a low level are not. The tumorigenic clones secrete transforming growth factor a (TGF‐α) in the culture medium whereas the non‐tumorigenic clones do not produce any detectable amount. Accordingly the level of TGF‐α mRNA is higher and the transcription rate of the gene is increased in the tumorigenic clones. The acquisition of the tumorigenic pheno‐type by cells of non‐tumorigenic clones, following introduction of c‐myc gene copies by transfection, is accompanied by an increase in the steady‐state level of TGF‐α mRNA. These findings suggest a role for an elevated level of TGF‐α production in the tumorigenic phenotype of SW613‐S cells. The possibility that this role is indirect is discussed. © 1992 Wiley‐Liss, Inc.
HLA‐B5‐restricted auto‐tumor‐specific cytotoxic T cells generated in mixed lymphocyte‐tumor‐cell culture
Authors: Wang P. , Végh Zs. , Vánky F. , Klein E.
Published: February 2007

T‐cell‐enriched lymphocyte populations derived from the malignant exudate of a patient with ovarian carcinoma were exposed to autologous tumor cells in the mixed lymphocyte‐tumor‐cell culture (MLTC) and propagated for 42 days. Proliferation of lymphocytes depended on exposures to autologous tumor cells and on the presence of IL‐2. After 7 days, the MLTC‐lymphocytes lysed K562 and the autologous tumor cells. The latter effect was not inhibited by monoclonal antibodies (MAbs) reactive with MHC class‐1 antigens or with CD3. After 7 restimulations, the culture was enriched in CD8+ cells (92%) and showed selective lytic activity against the autologous tumor. This function was inhibited by the α‐class 1 or α‐CD3 MAbs, and also by antibodies reactive with the HLA B locus or B5 allele products. The antibodies reactive with HLA A molecules had no such effect. It seems therefore that the function of the CTLs was restricted by HLA B5. Analysis of the TCRβ genes indicated clonal T‐cell expansion in this culture. This MLTC was I of 21 initiated with 11 blood‐and 10 tumor‐derived lymphocyte (TIL) populations prepared from the malignant effusions of ovarian carcinoma patients. None of these ex‐vivo lymphocytes lysed autologous tumor cells. In 17 MLTCs the lymphocytes did not proliferate, and in 3 cultures the proliferation was maintained only for 2–3 weeks. In 3 of 4 cultures auto‐tumor cytotoxicity was induced. © 1992 Wiley‐Liss, Inc.
Effect of smoking on folate levels in buccal mucosal cells
Authors: Piyathilake C. J. , Hine R. J. , Dasanayake A. P. , Richards E. W. , Freeberg L. E. , Vaughn W. H. , Krumdieck C. L.
Published: February 2007

The objective of the study was to document the existence of localized deficiency of folate in a tissue exposed to cigarette smoke, by analysis of oral and circulatory levels of this vitamin in smokers and non‐smokers. Buccal mucosal cells and blood samples were collected from 25 smokers and 34 non‐smokers. The Health Habits and History Questionnaire was completed by each subject. A 96‐well plate L. casei assay, along with preincubation with a folate‐free chick pancreas pteroyl‐gamma‐glutamyl hydrolase, was used to quantitate total buccal mucosal cell folates. The reproducibility (CV 5 to 7%) and recovery (95 to 106%) of the folate assay were satisfactory. Smokers had significantly lower buccal mucosal cell folate levels than did non‐smokers. The mean plasma folate level of smokers although within normal limits, was also significantly lower than that of non‐smokers. There were no significant differences in mean dietary folate intake or in alcohol consumption between the 2 groups. The strength of the positive association between smoking and plasma and buccal mucosal cell folate deficiency (by any definition) was moderate to strong and statistically significant. Our results indicate that cigarette smoking may result in a localized folate deficiency in buccal mucosal cells, independent of the plasma folate levels. © 1992 Wiley‐Liss, Inc.
Lung cancers associated with thorotrast exposure: High incidence of small‐cell carcinoma and implications for estimation of radon risk
Authors: Ishikawa Yuichi , Mori Takesaburo , Kato Yoshio , Tsuchiya Eiju , Machinami Rikuo , Sugano Haruo , Kitagawa Tomoyuki
Published: February 2007

The widely accepted concept that the alpha‐emitting radionuclide radon (222Rn,220Rn) induces lung cancers in humans has been based on the excess of lung tumours observed in underground miners extracting uranium or other substances. However, this poses the important question of whether radon is the only carcinogenic factor because such miners are also heavily exposed to mine dusts including silicates, diesel exhaust, etc. in their working environment. Patients to whom Thorotrast was administered continuously exhale radon (220Rn) derived from232Th deposits in the body and therefore provide a good model for lung carcinogenesis by radon without concomitant dust exposure. We therefore investigated lung‐cancer incidence in our epidemiological follow‐up series, analysing the histological types of 11 lung cancers which were found among 359 Thorotrast autopsy cases and measuring radioactivity in the breath of living Thorotrast patients. The study revealed that, while the proportion of small‐cell lung cancers considered to be related to alpha‐particles was significantly increased, the overall lung cancer incidence was not significantly higher than in controls, in spite of the high level of220Rn in the patients' breath. This result suggests that radon in the lung does induce cancers (particularly small‐cell carcinomas) but that the induction rate is not as high as expected from risk factors associated with mining. Thus, excess lung cancers among the miners might be related to the combined effects of exposure to radon and mine dusts, and not solely to radon. © 1992 Wiley‐Liss, Inc.
The gene encoding for MC56 determinant (drug‐sensitivity marker) is located on the short arm of human chromosome 11
Authors: Cianfriglia M. , Viora M. , Tombesi M. , Merendino N. , Esposito G. , Samoggia P. , Forsberg U. H. , Schröder J.
Published: February 2007

A panel of mouse × human B‐and T‐cell hybrids was analyzed for the expression of MC56 determinant which marks the drug‐sensitive state of CEM cells. Karyotypic and phenotypic analyses of the tested clones showed that the expression of MC56 determinant correlated to the presence of human chromosome 11 and segregated concordantly to the epitopes recognized by monoclonal antibodies in the CD44 cluster. By using a particular class of interspecific rodent × human‐cell hybrids in which the human genome counterpart is represented in the different clones only by human chromosome 11 or its fragments, we showed that the gene encoding for MC56 determinant is located on the region p 13‐pter of the short arm of chromosome 11. Therefore, the hypothesized homology between the drug‐sensitivity marker MC56 and the CD44 determinant is supported also by gene mapping studies. © 1992 Wiley‐Liss, Inc.
The lipoxygenase metabolite 12(S)‐hete promotes αllbβ3integrin‐mediated tumor‐cell spreading on fibronectin
Authors: Timar Jozsef , Chen Yong Q. , Liu Bin , Bazaz Rajesh , Taylor John D. , Honn Kenneth V.
Published: February 2007

Tumor‐cell interaction with the vessel wall during metastasis involves adhesion, induction of endothelial‐cell retraction and spreading on the exposed sub‐endothelial matrix. The signals for initiation of tumor‐cell spreading and the receptors involved are unknown. A protocol was developed to distinguish between initial tumor‐cell (B16 amelanotic melanoma; B16a) adhesion to and spreading on fibronectin. The time for maximum spreading was 50 min. Treatment with a lipoxygenase metabolite of arachidonic acid [12(S)‐HETE] resulted in maximum spreading in 15 min (max. effect approx. O.1 μM). Other lipoxygenase metabolites were ineffective. 12(S)‐HETE treatment induced a rearrangement of F‐actin, vinculin, vimentin intermediate filaments and integrin α11bβ3, but not integrin α5β1. Antibodies to α11bβ3 but not α5β1 blocked the 12(S)‐HETE effect on B16a spreading. B16a‐cell attachment to fibronectin resulted in increased metabolism of arachidonic acid to 12(S)‐HETE, which was inhibited by lipoxygenase but not by cyclo‐oxygenase inhibitors. Accordingly, lipoxygenase inhibitors but not cyclo‐oxygenase inhibitors blocked spontaneous B16a‐cell spreading. The protein‐kinase‐C inhibitors calphostin C, H7 and staurosporine also inhibited spreading, while the protein‐kinase‐A inhibitor H8 was ineffective. These data suggest that B16a‐cell spreading on fibronectin is initiated by a lipoxygenase metabolite [12(S)‐HETE] of arachidonic acid and is mediated by protein kinase C. © 1992 Wiley‐Liss, Inc.
An anti‐mucin immunotoxin BrE‐3‐ricin A‐chain is potently and selectively toxic to human small‐cell lung cancer
Authors: Derbyshire Elaine J. , Wawrzynczak Edward J.
Published: February 2007

Monoclonal antibodies (MAbs) known to recognize epithelial mucin or defined carbohydrate structures present on mucin molecules were screened for their ability to form cytotoxic agents with ricin A‐chain active against human small‐cell lung cancer (SCLC) in an indirect assay of immunotoxin cytotoxicity. Anti‐X hapten and anti‐Y hapten antibodies binding to a high proportion of SCLC cells mediated only weak to moderate effects on 3H‐leucine incorporation in combination with the screening agent, sheep anti‐mouse IgG F'ab‐ricin A‐chain. In contrast, the mouse MAb BrE‐3, recognizing the polypeptide core of the MUCI mucin gene product, exerted potent and selective cytotoxic effects in the assay. An immunotoxin made by the direct attachment of ricin A‐chain to BrE‐3 was selectively toxic to SCLC cell lines in tissue culture. The cytotoxic activity of BrE‐3‐ricin A‐chain was enhanced 100‐fold in the presence of monensin but not by lysosomotropic amines or calcium antagonists. Our findings suggest that anti‐mucin immunotoxins may have a therapeutic role to play in the treatment of SCLC. © 1992 Wiley‐Liss, Inc.
Immunotoxins directed against the high‐molecular‐weight melanoma‐associated antigen. Identification of potent antibody‐toxin combinations
Authors: Godal Aslak , Kumle Brita , Pihl Alexander , Juell Siri , Fodstad Øystein
Published: February 2007

To study factors influencing the cytotoxicity of immunotoxins (ITs), we compared the in vitro cytotoxicity of conjugates in which the plant toxin abrin and the bacterial toxin Pseudomonas exotoxin A (PE) were coupled by 2 different procedures to 2 MAbs, 9.2.27 and NR‐ML‐05, which bind to different epitopes on the melanoma‐associated antigen p250. The individual target cell lines differed widely in sensitivity to the different ITs, as assessed by measurement of protein synthesis inhibition. The action of the ITs was highly specific, as the toxicity of abrin and PE conjugates was respectively 20–540 and 2,200–550,000 times higher in antigen‐positive cell lines (FEMX, SESX, OHS) than in the antigen‐negative line KPDX. The PE conjugates prepared with the 2 different MAbs differed in potency by factors of 16–126 in the target‐cell lines, but were consistently more toxic than the abrin ITs. The results demonstrate that the cytotoxicity of ITs varies with the nature of both of its moieties and that optimal results require that toxins and MAbs be matched. Moreover, the 2 coupling procedures affected differentially the binding and potency of some ITs. Each of the 2 toxins was conjugated to a sheep anti‐mouse antibody (SAM) and the toxicity of these 2 conjugates was tested in an indirect approach using 9.2.27 and NR‐ML‐05 as primary MAbs. The results showed that the indirect procedure would have correctly predicted the most potent antibody‐toxin pair, indicating that the approach may be valid for selecting suitable combinations of MAbs and toxins for use as direct ITs. © 1992 Wiley‐Liss, Inc.
Prevention of programmed cell death in burkitt lymphoma cell lines by bcl‐2‐dependent and ‐independent mechanisms
Authors: Milner Anne E. , Johnson Gerald D. , Gregory Christopher D.
Published: February 2007

Burkitt lymphoma (BL) cell lines which retain the phenotypic characteristics of the freshly‐isolated tumour cells (group I cells) readily enter programmed cell death (apoptosis) in response to a variety of triggers. By contrast, isogenic BL cells which are phenotypically altered as a result of activation of their resident EBV genome (group‐III cells) are highly protected from apoptosis. Phenotypic changes in group‐III cells include the up‐regulation of the oncogene, bcl‐2. Expression of the 26‐kDa bcl‐2 protein in group‐1 BL cells following gene transfer was found to afford protection from apoptosis: the degree of protection was proportional to the amount of bcl‐2 protein expressed. When group‐1 bcl‐2 transfectants were compared with their group‐III counterparts it was found that, whilst bcl‐2 made a significant contribution in protecting from entry into apoptosis, hyper‐expression of bcl‐2 protein in group‐1 cells (well beyond that of group‐III cells) was necessary to attain the high levels of protection observed in group‐III cells. These results suggested that additional, bcl‐2‐independent, survival mechanisms could operate in BL cells. In support of this notion it was also found that: (I) prolonged culture of group‐1 lines in vitro resulted in enhanced survival in the absence of bcl‐2 up‐regulation, and (2) exposure of group‐1 cells to interferon‐α triggered a bcl‐2‐independent protective response. The molecular mechanisms of both the bcl‐2‐dependent and ‐independent survival pathways remain to be determined. © 1992 Wiley‐Liss, Inc.
Genes involved in tumor invasion and metastasis are differentially modulated by estradiol and progestin in human breast‐cancer cells
Authors: van den Brûle1 Frédéric A. , Engel Jason , Stetler‐Stevenson William G. , Liu Fu‐Tong , Sobel Mark E. , Castronovo Vincent
Published: February 2007

Invasion of basement membranes by cancer cells is a critical step in metastasis, which requires the coordinated expression of specific genes such as laminin receptors and metalloproteinases. Estradiol and progesterone modulate the clinical progression of steroid‐sensitive breast cancers; however, little is known about the molecular regulation of the invasive phenotype by these hormones. We therefore examined the effects of 10 nM estradiol and/or 10 nM progestin R5020 on the expression of 2 non‐integrin laminin binding proteins, the 67‐kDa laminin receptor (67LR) and HLBP31 as well as the 72‐kDa type‐IV collagenase (MMP‐2) and its inhibitor, TIMP‐2, in steroid‐receptorpositive (T47D and MCF‐7) and ‐negative (MDA‐MB 231) human breast‐cancer cells. The relative steady‐state level of 67LR mRNA was increased 2‐ to 3‐fold by estradiol in both MCF‐7 (p < 0.001) and T47D (p < 0.001) cells, also by R5020, alone or in combination with estradiol, in T47D cells (p < 0.001) and to a much less extent in MCF‐7 cells. HLBP31 mRNA and protein levels were increased 2‐ to 3‐fold (p < 0.001) by R5020 alone or in combination with estradiol. but not by estradiol alone. None of the steroid treatments affected the expression or activity of MMP‐2. Interestingly, however, TIMP‐2 mRNA levels and protein expression in MCF‐7 and T47D cells were 50% down‐regulated (p < 0.001) by treatment with R5020 or R5020 plus estradiol, but not by treatment with estradiol alone. None of these genes were modulated in steroid‐independent MDA‐MB231 cells. The data suggest that estradiol and progesterone might act as coordinators regulating specific genes in the steroid‐sensitive breast‐cancer cell, leading to the acquisition of the metastatic phenotype. © 1992 Wiley‐Liss, Inc.
The presence of concanavalin‐a(C0N‐A)‐like molecules on natural‐killer (NK)‐Sensitive target cells: Their possible role in swainsonine‐augmented human NK cytotoxicity
Authors: Yagita Masato , Noda Ichiro , Maehara Masanori , Fujieda Shigeharu , Inoue Yoshiko , Hoshino Takashi , Saksela Eero
Published: February 2007

In the present study we examined the expression of concanavalin‐A(Con‐A)‐like molecules on natural‐killer (NK)‐sensitive target cells and investigated their possible role in the human NK‐cell phenomenon. The incubation of either peripheral‐blood lymphocytes (PBL) or large granular lymphocytes (LGL) with swainsonine (SW), an inhibitor of mannosidase II, resulted in the augmentation of cytotoxicity against K562 leukemia cells. The enhanced cytotoxicity was associated with increased binding of fluorescein isothiocyanate‐conjugated Con‐A to SW‐treated effector cells, and immunofluorescence staining of the target K562 cells using goat anti‐Con‐A antibody (Ab) showed a significant positive shift in the flow cytometric pattern. Electrophoretic separation and immunoblotting analysis revealed that 4 components with a molecular weight of approximately 95, 80, 60 and 50 kDa were recognized by anti‐Con‐Ab from the detergent‐extract of K562 cells. The addition of Con‐A during the antibody incubation step of the Western blotting abolished their expression, thus excluding non‐specific binding of the antibody. The addition of Con‐A also strongly inhibited the cytotoxicity of SW‐treated effector cells (PBL or LGL) against K562 cells, and this inhibition was abolished by 40 mM α‐methyl‐mannopyranoside (α‐MM), which binds to Con‐A. Furthermore, Con‐A increased the binding frequency of SW‐treated LGL to K562. in spite of the inhibited cytotoxicity, and this effect could be neutralized by the further addition of α‐MM. Our results suggest that Con A‐like molecules might play an important role in cell‐cell interactions between SW‐treated effector cells and NK target cells. © 1992 Wiley‐Liss, Inc.
Variations in the ganglioside profile of uveal melanoma correlate with cytologic heterogeneity
Authors: Kanda Shinji , Cochran Alistair J. , Lee William R. , Morton Donald L. , Irie Reiko F.
Published: February 2007

Gangliosides may play an important role in the proliferation and spread of human malignant melanoma. Because the frequency of metastases in uveal and cutaneous melanoma differs, it is possible that they may express different gangliosides. We analyzed the ganglioside profiles of primary uveal melanoma in 14 cases and of cutaneous melanoma metastasis in 19 cases. In cutaneous melanoma, GM3 ranged from 4.2% to 74.6% and GD3 from 22.1% to 91.8% of total lipid‐bound sialic acid. GM2 (found in 13 of 19 cases, ranging from 0.5% to 11.7%), GD2 (11/19, 0.5%–22.0%) and 9‐O‐acetyl‐GD3 (13/19, 0.5%‐12.6%) were also frequently observed. By contrast, in 11 cases of uveal melanoma, GM3 was >90%, GD3 was < 10%, GM2 was <1.1%; neither GD2 nor 9‐O‐acetyl‐GD3 were detected. The ganglioside profiles of these uveal melanomas were virtually identical to those of normal melanocytes obtained from foreskins. Histological examination of these 11 biopsies showed a monomorphous cell composition, but neither infiltration of lymphocytes or melanophages nor cell necrosis was observed. In 3 other cases, GD3 was increased to 19.5%‐46.0%. Histological examination of these 3 biopsy specimens showed at least 2 populations of tumor cells that were separable based on morphological grounds, and mononuclear inflammatory cells interspersed among the tumor cells. An increase in GD3 appears to be related to tumor polyclonality and infiltration of the tumor by lymphocytes and macrophages. These results suggest that ganglioside expression of uveal melanoma is associated with host immune responses to the tumor. Furthermore, the low metastatic capacity of uveal melanoma, in contrast to the high metastatic rate of cutaneous melanoma, may be a result of its differentiated ganglioside expression, which is strikingly similar to that of normal melanocytes. © 1992 Wiley‐Liss, Inc.
Expression of different tenascin isoforms in normal, hyperplastic and neoplastic human breast tissues
Authors: Borsi Laura , Carnemolla Barbara , Nicolò Guido , Spina Bruno , Tanara Giorgio , Zardi Luciano
Published: February 2007

Functionally different tenascin (TN) isoforms, containing varying numbers of a 91 amino‐acid motif resembling the fibronectin type‐III homology repeat, may be generated by alternative splicing of the TN primary transcript. In fact, only the TN isoform containing the alternatively spliced region can induce loss of focal adhesion in cultured cells and seems to be able to facilitate cell migration. We examined the patterns of alternative splicing of the TN primary transcript in normal, hyperplastic and neoplastic breast tissues, and found that, in all the invasive breast carcinomas analyzed, the relative amount of TN mRNA in which the alternatively spliced region was included was about 10 times higher than in RNA from normal breast tissues. A similar result was observed in phyllodes tumors and in those fibroadenomas which showed very high stromal cellularity. Western‐blot analysis using different monoclonal antibodies showed the same pattern as that seen in Northern blotting. The data reported here suggest that, in the breast, expression of the high‐molecular‐mass TN isoform is a marker of stromal element proliferation and that, in invasive breast carcinomas, this TN isoform could play a role in generating a permissive environment for proliferation, invasion and metastasis of neoplastic epithelial cells. © 1992 Wiley‐Liss, Inc.
Effect of DNA ploidy classification on prognosis in breast cancer
Authors: Joensuu Heikki , Alanen Kalle , Falkmer Ursula G. , Klemi Pekka , Nordling Stig , Remvikos Yorghos , Toikkanen Sakari
Published: February 2007

A series of 327 breast cancers was analyzed for DNA ploidy by flow cytometry from paraffin‐embedded tissue, and the resulting DNA histograms were classified independently by 6 researchers in the field as DNA diploid (Di), aneuploid (An), tetraploid (Te), multiploid (Mu), or technically uninterpretable. The frequency of diploid, aneuploid, tetraploid and multiploid cancers varied from 28 to 41%, 33 to 49%, 8 to 21% and 2 to 6%. According to the scale Di‐An‐Te‐Mu, DNA ploidy was not significantly associated with breast‐cancer mortality by 2 classifiers, but if DNA euploid cancers (Di+Te) were tested against non‐euploid, or diploid cancers against non‐diploid, all classifiers found DNA euploid and diploid cancers to have better prognosis. Mortality associated with diploid or tetraploid cancers decreased with improving histogram quality and increasing uniformity of classification, whereas that associated with aneuploid cancers remained unaltered. Among the cases where all classifiers agreed on ploidy, tetraploid, diploid and aneuploid cancers were associated with 100%, 88% and 68% 5‐year survival rates. In this sub‐set the S‐phase fraction and possibly DNA ploidy were independent prognostic factors, together with histological grade, axillary node status, and primary tumor size. © 1992 Wiley‐Liss, Inc.
Correlation between the serum level of hepatitis C virus RNA and disease activities in acute and chronic hepatitis C
Authors: Gunji Toshiaki , Kato Nobuyuki , Mori Shigehisa , Ootsuyama Yuko , Hijikata Makoto , Imawari Michio , Shimotohno Kunitada
Published: February 2007

The influence of viremia on hepatic injury in patients infected with hepatitis C virus was examined by analysis of the relationship between alanine aminotransferase activity and the amount of hepatitis C virus RNA in sequential serum samples from 1 untreated patient with acute hepatitis C and 3 untreated patients with chronic hepatitis C. Semiquantitative analysis by the competitive‐reverse‐transcription/polymerase‐chain‐reaction method indicated that the quantity of hepatitis C virus RNA in the serum affected the disease activities of acute and chronic hepatitis C through their natural clinical courses in all these patients. The nucleotide sequence encoding the putative envelope region of the viral genome in the patient with acute hepatitis C was examined. Blood samples taken serially at 2 times of exacerbation of the hepatitis revealed 2 nucleotide mutations, resulting in changes of predicted amino acid residues. This finding suggests that nucleotide mutations in the envelope region of the viral genome may be responsible for the recurrent hepatic injury attributed to recurrence of viremia in patients with hepatitis C. From these aspects, the serial divergence of the virus genome in infected individuals, especially in the region encoding the viral envelope protein, may possibly play an important role in developing chronic infection of hepatitis C virus. © 1992 Wiley‐Liss, Inc.
Interaction of gangliosides with the methyltransferase‐I of microsomes from normal and neoplastic human mammary gland
Authors: Losa Gabriele A. , Graber Riccardo , Chigorno Vanna , Sonnino Sandro
Published: February 2007

The activity of the phospholipid methyltransferase‐1, which catalyzes the rapid transmethylation of phosphatidylethanolamine to phosphatidylcholine, was found to be about 6‐fold enhanced in microsomal membranes of breast cancer with respect to the level found in normal human mammary gland. Exogenous gangliosides GM1 and GM2 added to neoplastic breast microsomes induced progressive inhibition of the methyltransferase activity. In contrast, in microsomal membranes of non‐neoplastic breast tissue treated with these gangliosides, the methyltransferase activity was markedly increased. The addition of cholesterol to these microsomes led to complete inhibition of the GM1 ‐stimulated enzyme activity. The methyltransferase activity was not affected by GM3 alone in either type of tissue. Experiments carried out on non‐neoplastic microsomes revealed that the phospholipid methyltransferase‐1 was affected by that portion of gangliosides which remained stable associated to microsomal membranes. © 1992 Wiley‐Liss, Inc.