bluuebunny/crossref_metadata_2025 huggingface dataset:
Selective Killing of Transformed Fibroblasts by Combined Treatment with Cycloheximide and Aphidicolin
Authors: Okuda Atsuyuki , Kimura Genki
Published: August 2005

The possibility of selective killing of transformed cells in a mixed population of untransformed and transformed cells was examined using a cell culture system of rat 3Y1 fibroblasts (parental 3Y1 cells, 3Y1 cells transformed with either SV40, polyoma virus, Rous avian sarcoma virus, E1A gene of adenovirus type 12, or H‐v‐ras oncogene). The principle of the selective killing is as follows. Under suboptimal culture conditions, untransformed cells are inhibited from progressing through G1 phase and retain viability, while transformed cells are not arrested. When DNA synthesis is inhibited for a long period, both types of cells in S phase die. Therefore, if we administer inhibitors of G1 progression and of DNA synthesis simultaneously to a cell population consisting of untransformed and transformed cells, most untransformed cells are arrested in G1 phase, retaining viability, while transformed cells leak from the G1 phase, cease DNA synthesis, and gradually die. The present study shows that all types of transformants in stationary‐phase cultures (consisting of cells mainly with a G1 DNA content) were killed to higher extents compared with untransformed cells, during incubation at lower cell densities with a combination of cycloheximide (G1 inhibitor) and aphidicolin (DNA‐synthesis inhibitor). However, cycloheximide reduced the killing effect of aphidicolin by changing the irreversible DNA‐synthesis inhibition to a reversible inhibition. The availability of G1 inhibitors that do not interfere with the irreversibility of inhibition of DNA synthesis is required for the treatment of cancer based on this idea.
Immunocytochemical Identification and Localization of the Mr 22,000 Calcium‐binding Protein (Sorcin) in an Adriamycin‐resistant Myelogenous Leukemia Cell Line
Authors: Sugawara Isamu , Mizumoto Kenji , Ohkochi Etsuko , Hamada Hirofumi , Tsuruo Takashi , Mori Shigeo
Published: August 2005

Monoclonal antibody against the Mr 22,000 calcium‐binding protein (sorcin) from an adriamycin‐resistant myelogenous leukemia cell line K562 (K562/ADM) was prepared and used as a probe to study the localization of sorcin in K562/ADM cells and the parental cell line, K562. Analysis of extracts from K562/ADM cells by sodium dodecyl sulfate‐polyacrylamide gel electrophoresis and fluorescence image analysis showed that K562/ADM cells possessed abundant sorcin in the cytoplasm which was almost entirely absent from the drug‐sensitive parental cell line, K562. Furthermore, immuno‐electron microscopic studies revealed that sorcin was closely associated with free ribosomes, rough endoplasmic reticulum, mitochondria, microfilament bundles and perinuclear membranes. These observations provide the first clue that the Ca‐binding protein, sorcin, may play an important role in the development of the multidrug resistance phenomenon, although the relationship between sorcin and P‐glycoprotein is still unknown.
Functional Analysis of Mononuclear Cells Infiltrating into Tumors: Establishment of a New System to Obtain Functionally Active Tumor‐infiltrating Cells
Authors: Shijubo Noriharu , Uede Toshimitsu , Nomura Naohiro , Kikuchi Kokichi
Published: August 2005

We have demonstrated that tumor‐infiltrating T cells play a crucial role in tumor growth. In order to investigate the precise function of tumor‐infiltrating cells, we avoided the use of any enzyme for separation of tumor‐infiltrating cells from the solid tumor mass and established a new system in which we could easily obtain functionally active tumor‐infiltrating cells in large amounts from the peritoneal cavity of T‐9 gliosarcoma‐sensitized rats. In this system, viable T‐9 cells were intraperitoneally administered and peritoneal exudate cells were recovered. Importantly, cells obtained from the intraperitoneal cavity are phenotypically and functionally indistinguishable from those obtained from subcutaneous tumor masses. Furthermore, injection of purified lymphocyte migration factor‐b resulted in the accumulation of CD8 (+) cytotoxic T cells in the peritoneal cavity. Therefore, this system also provides a means by which the in vivo function of chemotactic factor can be tested.
Enhancement of the Antibody‐dependent Tumoricidal Activity of Human Monocytes by Human Monocytic Colony‐stimulating Factor
Authors: Suzu Shinya , Yanai Nobuya , Saito Minoru , Kawashima Takuji , Kasahara Tadashi , Saito Masaki , Takaku Fumimaro , Motoyoshi Kazuo
Published: August 2005

Human monocytic colony‐stimulating factor (hM‐CSF) stimulates mature cells as well as progenitor cells of human monocyte lineage. In the present study, we investigated the effect of completely purified native hM‐CSF on the tumoricidal activity of human monocytes against Raji (Burkitt lymphoma‐derived cell line) using a chromium release assay. Preincubation of human peripheral blood monocytes with hM‐CSF for 48 h markedly enhanced their antibody‐dependent cellular cytotoxicity (ADCC). This enhancing effect was dependent on the concentration of hM‐CSF, and maximal enhancement was achieved at the concentration of 66 ng/ml. The number of receptor sites on monocytes against the Fc portion of immunoglobulin G increased 2‐ to 4‐fold during 48 h incubation with hM‐CSF. These results suggest that hM‐CSF stimulates monocyte‐ADCC by increasing the number of Fc receptor sites on human monocytes.
Proposed Distal Margin for Resection of Rectal Cancer
Authors: Kameda Kanji , Furusawa Motonosuke , Mori Masaki , Sugimachi Keizo
Published: August 2005

To determine the adequate distal margin, particularly from the point of extent of lymph node metastasis, 2,333 lymph nodes from 44 patients with rectal carcinoma were evaluated, using a clearing method. The tumors were divided into two growth patterns; infiltrative and localized. Lymph node metastasis was histo‐pathologically examined with special attention focused on nodes on the distal side of the tumor. The intramural spread was also given attention. The proximal pararectal lymph nodes often contained malignant cells, whereas the distal ones were rarely involved, and if involved, they were present within 1 cm from the tumor. Pararectal lymph node metastasis and intramural spread were seen with a range of 1 cm and 0.5 cm, respectively, in the localized type and 1 cm and 2.1 cm, respectively in the infiltrative type. Based on these findings, the distal margin for surgical resection of rectal carcinoma is considered to be 2 cm for the localized type and 3 cm for the infiltrative type.
Point Mutation of the neu Gene in Rat Neural Tumor RT4‐AC Cells: Suppression of Tumorigenicity by s‐Myc
Authors: Lee Soo‐Yong , Sugiyama Akinori , Sueoka Noboru , Kuchino Yoshiyuki
Published: August 2005

Forced expression of the s‐myc gene suppressed the tumorigenicity of rat RT4‐AC tumor cells in nude mice, as reported previously. Polymerase chain reaction (PCR) analysis indicated that RT4‐AC cells established from a tumor of the peripheral nervous system contain an activated neu gene with a T→A transversion in the transmembrane domain. Synthesis of a protein of 60 kd in RT4‐AC cells was specifically inhibited by expression of the s‐myc gene. These results strongly suggest that s‐Myc suppresses the transforming activity of rat neural cells transformed by expression of the activated neu gene, and plays an important role in regulating expression of a cellular gene contributing to cell transformation, such as the gene that encodes the p60 protein.
A Comparative Case‐Control Study of Colorectal Cancer and Adenoma
Authors: Kato Ikuko , Tominaga Suketami , Matsuura Akira , Yoshii Yuri , Shirai Masato , Kobayashi Seibi
Published: August 2005

We conducted a comparative case‐control study of colorectal cancer and adenoma involving 221 cases with colorectal cancer, 525 cases with colorectal adenoma and 578 neighborhood controls. Daily vegetables intake was associated with lower risks of distal colon adenoma (relative risk (RR)=0.59, 95% confidence interval (CI): 0.39–0.89) and rectal cancer (RR=0.46, 95% CI: 0.25–0.84). Daily beans intake was associated with lower risk of colon adenoma (RR=0.58, 95% CI: 0.37–0.91 for the proximal colon and RR=0.63, 95% CI: 0.45–0.88 for the distal colon) and daily intake of seaweeds was associated with lower risk of rectal cancer (RR=0.42, 95% CI: 0.22–0.82). Daily intake of fish and shellfish also showed an inverse association with the risk of colon adenoma (RR=0.67, 95% CI: 0.45–0.99 for the proximal colon and RR=0.70, 95% CI: 0.52–0.94 for the distal colon). Generally, intakes of animal or vegetable fat‐rich foods, especially meats, were associated with decreases in risks of both adenoma and cancer, though the association of cancer was not statistically significant. Other than dietary factors, daily alcohol drinking was associated with an increased risk of adenoma in the proximal colon (RR=1.95, 95% CI: 1.15–3.29) and ex‐drinkers showed higher risks for colon adenoma and colorectal cancer. Sports or occupational activities and coffee drinking were inversely associated and family history of colorectal cancer was positively associated with the risks of both colorectal adenoma and cancer.
Analysis of Stomach Cancer Incidence by Histologic Subtypes Based on a Mathematical Model of Multistage Cancer Induction and Exponential Growth
Authors: Yamaguchi Naohito , Watanabe Shaw , Maruyama Keiichi , Okubo Toshiteru
Published: August 2005

A mathematical model incorporating the processes of both cancer induction and subsequent tumor growth has been developed. The model was applied to incidence data of stomach cancer classified into histologic subtypes: papillary adenocarcinoma (PAP), well and moderately differentiated tubular adenocarcinomas (WEL and MOD), poorly differentiated adenocarcinoma (POR), mucinous adenocarcinoma (MUC) and signet‐ring cell carcinoma (SIG). The multistage theory was assumed for cancer induction as in the Armitage‐Doll model. For the period of growth, exponential growth was assumed and clinical surfacing was formulated as a stochastic process related to tumor diameter. The number of stages in cancer induction and the tumor growth rate were simultaneously estimated for each histologic subtype using the maximum likelihood procedure. The present model showed better fits than the Armitage‐Doll model in most histologic subtypes except WEL. PAP, WEL and MOD, which are characterized as differentiated subtypes with less mucous production, showed different features from POR, MUC and SIG: 1) the number of stages was estimated to be larger, 2) the differences in incidence rates between males and females were more marked, and 3) males tended to have larger growth rates in PAP and MOD, while in POR, MUC and SIG, females had larger values. The present study showed that an analysis by histologic subtypes is of importance in stomach cancer and that the period of tumor growth should not be ignored when formulating a model of the natural history of stomach cancer.
Repressive Effect of the nef cDNA of Human Immunodeficiency Virus Type 1 on the Promoter Activity of the Viral Long Terminal Repeat
Authors: Mori Shigehisa , Takada Ritsuko , Shimotohno Kunitada , Okamoto Takashi
Published: August 2005

The nef gene product of human immunodeficiency virus type 1 (HIV‐1) has been implicated as a negative factor for viral replication and is suspected to play an important role in the maintenance of viral latency. However, there seems to be evidence both for and against the negative effect of nef gene product. In the present report, we reevaluated the function of the nef gene by means of transient CAT assays with two human T cell lines. In most of the experiments, carefully controlled triplicate studies were carried out. We observed that not only the nef‐expression plasmid, but also an effector plasmid containing the nef cDNA sequence in a reverse orientation, not expressing the Nef protein, showed a similar extent of repression of the HIV‐1 promoter activity. We also examined the repressive effect of the nef cDNA with deletion mutants of HIV‐1 long terminal repeat and heterologous promoters. The results led us to conclude that the apparent “repressor”‐like action of the nef cDNA itself could be explained by competition for certain transcription factors required for HIV‐1 gene expression by identical sequences also present in the nef cDNA.
Heterogeneity of Proliferative Activity in Nodule‐in‐Nodule Lesions of Small Hepatocellular Carcinoma
Authors: Matsuno Yoshihiro , Hirohashi Setsuo , Furuya Shuichiroh , Sakamoto Michiie , Mukai Kiyoshi , Shimosato Yukio
Published: August 2005

Surgically resected small hepatocellular carcinomas showing “nodule‐in‐nodule’formation were analyzed in terms of cell proliferative activity. The analysis was achieved by successful immunohistochemical demonstration of proliferating cell nuclear antigen in formalin‐fixed paraffin‐embedded tissue sections. Eight nodules (up to 3 cm in diameter) examined were either atypical adenomatous hyperplasia or hepatocellular carcinoma of low histologic grade, containing a discrete inner nodular area composed of obvious hepatocellular carcinoma of higher histologic grade. In all cases, the proliferating cell nuclear antigen labeling index of the latter area was much higher than that of the former, which in turn was slightly higher than that of the non‐cancerous liver of the patient in 6 cases. The data presented here provide supporting evidence that the successive emergence and expansion of a more rapidly proliferating subclone within a nodule result in the stepwise progression of malignancy of human hepatocellular carcinoma.
An Antibody‐Tumor Model for the Targeting of CA125‐producing Gynecologic Malignancies
Authors: Saga Tsuneo , Ishiwata Isamu , Endo Keigo , Sakahara Harumi , Koizumi Mitsuru , Watanabe Yuji , Nakai Toshiharu , Hosono Makoto , Ishikawa Hiroshi , Sawada Masumi , Konishi Junji
Published: August 2005

By immunizing a mouse with HOUA‐1 cells established from an endometrial cancer patient, two murine monoclonal antibodies designated 196–14 and 196–28 were generated, which were reactive with ovarian cancer‐associated antigen CA125, originally defined by OC125 antibody. Antigenic determinants of these antibodies, although overlapping each other, were different from that of OC125 and the combined use of 125I‐labeled 196–14 and OC125‐coated beads markedly increased the sensitivity of measuring CA125 antigen. Both radioiodinated and 111In‐labeled 196–14 localized well in CA125‐producing human ovarian cancer tissues OVA‐5 xenografted in nude mice. The biodistribution of radioiodinated 196–14 was quite different from that of 111In‐labeled 196–14. Radioiodine was cleared faster from the OVA‐5 tumor, making a clear contrast to the prolonged retention of 111In in the tumor. Initial tumor uptake of radioiodinated 196–14 was the same as that of 111In‐labeled 196–14 but decreased thereafter, due to the dehalogenation of radioiodinated antibody in the tumor. This antibody‐tumor model seems to be suitable for examining the usefulness of monoclonal antibody‐conjugates in the diagnosis and therapy of CA125‐producing endometrial or ovarian cancers.
Monoclonal Anti‐P‐glycoprotein Antibody‐dependent Killing of Multidrug‐resistant Tumor Cells by Human Mononuclear Cells
Authors: Heike Yuji , Hamada Hirofumi , Inamura Noriaki , Sone Saburo , Ogura Takeshi , Tsuruo Takashi
Published: August 2005

Mouse monoclonal antibodies (MRK16 and MRK17) against human multidrug‐resistant cancer cell lines were tested for antibody‐dependent cytotoxicity mediated by human blood mononuclear cells, using a 4‐h 51Cr release assay. MRK16 (IgG28 isotype) was shown to be more effective than MRK17 (IgG1 isotype). Moreover, when four pairs of drug‐resistant and their parent sensitive human cancer cells were tested for antibody‐dependent cell‐mediated cytolysis (ADCC) using MRK16, only the drug‐resistant cell lines were susceptible to ADCC reaction. When highly purified lymphocytes (>99%) and monocytes (>97%) were isolated from blood mononuclear cells by centrifugal elutriation and adherence, MRK16 promoted both lymphocyte‐ and monocyte‐mediated tumor cell killing, whereas MRK17 induced only a lymphocyte‐mediated ADCC reaction. These results suggest that MRK16 of IgG28 subtype may be a useful therapeutic agent in eradication of drug‐resistant cancer cells expressing P‐glycoprotein through ADCC reaction.
Antitumor Activity of Recombinant Human Interleukin‐1 against Heterotransplanted Human Non‐Hodgkin Lymphomas in Nude Mice
Authors: Miyamoto Tadaaki , Wu Shu‐guang
Published: August 2005

Antitumor activity of recombinant human interleukin 1α (IL‐1) against seven human non‐Hodgkin lymphomas grown in athymic nude mice was studied. Growth of the lymphomas was markedly inhibited after an injection of 0.4 mg/kg IL‐1. The growth inhibition of Burkitt lymphoma was found to be dose‐dependent up to 0.4 mg/kg, reaching a plateau thereafter. The loss of colony‐forming ability of the cells and the loss of cell viability showed the same type of dose‐dependence and progressed during 24 h following an injection of IL‐1. In accordance with these observations, histopathologic examination revealed progressively spreading coagulative necrosis without bleeding. Little infiltration of inflammatory cells into the tumor tissue was observed. IL‐1 growth inhibition of T lymphoma in beige nude mice having low natural killer activity was similar to that in nude mice. These findings suggested that the antitumor effects might not be produced through cell‐mediated antitumor actions. Immunocytological examination with anti‐IL‐1 antibody revealed that administered IL‐1 was bound to the lymphoma cells, suggesting that IL‐1 receptor is probably expressed on these cells in vivo. The antitumor action of IL‐1 on the lymphomas may be exerted directly through the IL‐1 receptor.
Prediction from Creatinine Clearance of Thrombocytopenia and Recommended Dose in Patients Receiving (Glycolato‐O, O′)‐diammine Platinum (II) (NSC 375101D)
Authors: Sasaki Yasutsuna , Fukuda Masaaki , Morita Masashige , Shinkai Tetsu , Eguchi Kenji , Tamura Tomohide , Ohe Yuichiro , Yamada Kohzo , Kojima Akira , Nakagawa Kazuhiko , Ohmori Toru , Saijo Nagahiro
Published: August 2005

Thrombocytopenia is the most serious dose‐limiting toxicity of (glycolato‐O, O′)‐diammine platinum (II) (254‐S; NSC 375101D), one of the new platinum analogues. A total of 38 patients treated with 100 mg of 254‐S per m2 in a clinical phase II study were retrospectively analyzed to determine the factors influencing thrombocytopenia. Performance status, sex, age and prior chemotherapy did not affect either the percent reduction of platelets or the nadir platelet count. However, creatinine clearance (Ccr.: ml/min) was found to be a predictive variable for thrombocytopenia. There was a significant relationship between nadir platelet count and Ccr. (R2=0.637) or relative dose [RD = (dose of 254‐S: mg/m2)/Ccr.] (R2= 0.707). From these observations, the following equations for predicting nadir platelet count associated with administration of 254‐S were proposed.

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If we administer 100 mg of 254‐S per m2 to a patient with Ccr. of less than 40.3 ml/min, thrombocytopenia of grade 3 or more according to the Eastern Cooperative Oncology Group (ECOG) criteria (< 50,000/mm3) can be predicted from equation (A). In these patients, dose modification using equation (B) is recommended.
Effect of Butylated Hydroxyanisole on the Level of DNA Adduction by Aristolochic Acid in the Rat Forestomach and Liver
Authors: Routledge Michael N. , Orton Terry C. , Lord Peter G. , Garner R. Colin
Published: August 2005

Administration of butylated hydroxyanisole (BHA) orally at either 0.5 g or 1 g/kg daily for 14 days to rats did not produce any DNA adducts in the forestomach as measured by the 32P‐postlabeling method using (1) limiting concentrations of 32P‐ATP; (2) nuclease P1 enhancement; or (3) butanol extraction. Experiments were conducted to establish the effects of BHA administration on aristolochic acid (AA) DNA adduct formation in the forestomach and liver, when BHA was administered prior to, together with or after AA administration. Adduct levels per 109 nucleotides in the liver after oral dosing daily for 5 days with 1 mg/kg AA and BHA (1 g/kg) or corn oil (5 ml/kg) for 7 days were as follows: (a) BHA and AA given simultaneously; 235±71, (b) AA+corn oil; 63±39, (c) AA followed by BHA; 57±13, (d) AA followed by corn oil; 91±38, (e) BHA followed by AA; 90±12, (f) corn oil followed by AA; 83±24. For the forestomach the values were: (a) 236±86, (b) 77±25, (c) 367±97, (d) 296±47, (e) 217±81, (f) 70±64. These data suggest that BHA could have an enhancing effect on AA‐induced lesions in the forestomach if dosed together with, or prior to, AA as adduct levels are significantly higher than in controls.
Amplification of HTLV‐1‐related Sequences among Patients with Neurological Disorders in Highly Endemic Nagasaki: Lack of Evidence for Association of HTLV‐1 with Multiple Sclerosis
Authors: Nakayama Daisuke , Katamine Shigeru , Kanazawa Hajime , Shibuya Noritoshi , Kawase Ken'ichiro , Moriuchi Ryozo , Miyamoto Tsutomu , Hino Shigeo
Published: August 2005

We studied DNA sequences homologous to HTLV‐1 in peripheral blood mononuclear cells (PBMC) of 30 patients with neurological disorders in Kyushu, where HTLV‐1 is highly endemic. The regions of HTLV‐1 amplified by means of polymerase chain reaction (PCR) included U3, U5, gag, pol, env and pX. Our system specifically detected HTLV‐1 sequences only from HTLV‐1‐positive cells and was sufficiently sensitive to detect one proviral copy per 103 PBMC. All PCR‐positive cases were seropositive, including 4 cases of tropical spastic paraparesis/HTLV‐1‐associated myelopathy (TSP/HAM) (4/4), and one case each of myasthenia gravis (1/7) and multiple sclerosis (1/8). The PCR‐positive rate of patients, excluding 4 TSP/HAM cases, was 8% (2/26), which is similar to the seroprevalence of the adult population in the area. The data suggest that multiple sclerosis is not associated with prototype HTLV‐1.
Decrease in the Metabolic Activating Capacities of Arylamines in Livers Bearing Hyperplastic Nodules: Association with the Selective Changes in Hepatic P‐450 Isozymes
Authors: Ozawa Shogo , Abu‐Zeid Medhat , Murayama Norie , Yamazoe Yasushi , Kato Ryuichi
Published: August 2005

The mechanism of the alteration in carcinogenic arylamine‐activating capacities in livers bearing pre‐neoplastic (or hyperplastic) nodules induced by the Solt‐Farber protocol was investigated in relation to the changes in hepatic cytochrome P‐450 isozymes. In the Salmonella mutagenesis test, the numbers of revertants induced with 2‐amino‐3‐methylimidazo[4,5‐f]quinoline and 2‐aminofluorene were significantly lower in the presence of microsomes of nodule‐bearing livers than of control livers. A similar tendency was also observed with another heterocyclic arylamine, 2‐amino‐6‐methyldipyrido‐[1,2‐a:3′,2′‐d]imidazole. In Western blots using specific antibodies against 5 different forms of cytochrome P‐450, hepatic contents of P‐450‐male (a main constitutive form) and P‐450b (a main phenobarbital‐inducible form) were decreased in the livers with hyperplastic nodules to 63% and 35% of the corresponding controls, while no significant decrease was observed in the contents of P‐448‐H (a main 3‐methylcholanthrene‐inducible form), P‐4506β‐1 (testosterone 6β‐hydroxylase) and P‐450e (a phenobarbital‐inducible form). In accordance with the reduction in P‐450‐male, capacities for microsomal 16α‐ and 2α‐hydroxylations, but not 6β‐hydroxylation, of testosterone were decreased in the livers with hyperplastic nodules. Although P‐448‐H has higher capacities for the activation of arylamines than does P‐450‐male, the hepatic content of P‐450‐male is more than ten‐fold higher than that of P‐448‐H in both normal and nodule‐bearing livers. These results indicate that the selective decrease in hepatic content of P‐450‐male is likely to be a main cause of the decrease in arylamine metabolic activating capacities in livers with hyperplastic nodules.
Cyclophosphamide‐dependent Lymph Node Modification in Lymph Node Metastasis of MM48 Tumor Cells in Syngeneic Mice
Authors: Yoshida Kikuyoshi , Tachibana Takehiko
Published: August 2005

We investigated the role of immunosuppressive activity induced in the regional lymph nodes (RLN, popliteal lymph nodes) in the establishment of lymph node metastasis by cyclophosphamide (CY) administration. The CY treatment led to the elimination of suppressive activity with the appearance of positive immune responses, and the inhibition of lymph node metastasis of MM48 tumor cells. In CY‐treated mice, the removal of RLN together with the primary tumor lowered the survival rate compared with the mice in which the RLN remained intact. During 4 days after primary tumor resection, the proliferation of tumor cells in the RLN was significantly decreased in CY‐treated mice. These results suggested that the induction of suppressive activity in the lymph node is closely associated with the establishment of lymph node metastasis.
In vivo Analysis of Tumor Vascularization in the Rat
Authors: Hori Katsuyoshi , Suzuki Maroh , Tanda Shigeru , Saito Sachiko
Published: August 2005

By using transparent chambers in rats, we have directly observed tumor‐induced neovascularization in the early stage and the formation of intricate networks in Yoshida rat ascites hepatoma AH109A and Sato lung carcinoma at high magnification. We counted branching point numbers per unit area in the microvascular network with and without tumors in order to clarify the sites from which new vascular sprouts originate. Branching point number per unit area in normal tissue was 13.6 ± 7.4/0.1 mm2 in the field near a terminal arteriole, and 12.9 ± 7.3/0.1 mm2 in the field distant from a terminal arteriole. There was no significant difference between these two fields in the normal vascular network. On the other hand, in the tumor vascular network, the branching point number in the field near a terminal arteriole was 50.4 ± 12.6/0.1 mm2, and 30.1 ± 11.5/0.1 mm2 in the field distant from a terminal arteriole. The difference is highly significant (P<0.001). The frequency with which new capillaries originated from veins and venules was very low. We concluded from these results that the position from which tumor vessels originated was usually the terminal portion of a terminal arteriole.
Cytocidal Activity of a Synthetic Isoprenoid, N‐Solanesyl‐N, N′‐bis(3,4‐dimethoxy‐benzyl)ethylenediamine, and Its Potentiation of Antitumor Drugs against Multidrug‐resistant and Sensitive Cells in vitro
Authors: Suzuki Hideo , Tomida Akihiro , Nishimura Toshio
Published: August 2005

A synthetic isoprenoid, N‐solanesyl‐N, N′‐bis(3,4‐dimethoxybenzyl)ethylenediamine (SDB‐ethylene‐diamine), inhibited the colony formation of multidrug‐resistant mutant cell lines derived from Chinese hamster V79 (V79/ADM) and human hepatoma PLC/PRF/5 (PLC/COL) cells to a greater extent than that of the parental cells. When combined with other clinically useful antitumor agents, it potentiated the cytotoxic activity of almost all kinds of drugs tested including adriamycin (ADM), actinomycin D, vincristine, cytosine arabinoside, and 5‐fluorouracil (5‐FU), and the potentiation ratios were higher against V79/ADM cells than against V79/S cells. Among the antitumor agents tested, the activities of bleomycin‐group antibiotics were more strongly enhanced by SDB‐ethylenediamine and the potentiation was higher in the parental cells than in V79/ADM cells. SDB‐ethylenediamine enhanced the uptake of ADM and daunorubicin into V79/ADM and its parental cells, but it did not increase the uptake of 5‐FU or peplomycin, indicating that different mechanisms operate for potentiation in the cases of the latter drugs, i.e., not simply an increase of intracellular drug uptake. Two fragments of SDB‐ethylenediamine, solanesol (polyprenoid moiety) and the diamine component (verapamil‐like moiety), showed neither cytotoxic activity nor potentiator activity, even if they were mixed together, indicating that the steric conformation of intact SDB‐ethylenediamine molecule is important for these two activities.
Synergistic Effect of Ultrasound and Hematoporphyrin on Sarcoma 180
Authors: Yumita Nagahiko , Nishigaki Ryuichiro , Umemura Koshiro , Umemura Shin‐ichiro
Published: August 2005

The antitumor effects of combined use of ultrasound (US) and a photosensitizer, hematoporphyrin (Hp), were determined in mice bearing sarcoma 180. In order to find the optimum timing of the US irradiation after the administration of Hp, the Hp concentrations in the tumor and in the plasma were determined and were analyzed pharmacokinetically. Antitumor effects were evaluated by measuring the tumor size and the tumor weight. Hp alone showed no antitumor effect but US alone showed a slight antitumor effect. The combined treatment with US and Hp showed marked synergistic effects on sarcoma 180 (inhibition ratio was 74% of the control). From these results, the enhancement of antitumor effect is thought to be caused by the sensitization of tumor cells to US mediated by Hp.
An Unusual Ring Contraction in the Formation of N‐Nitrosohexamethyleneimine and N‐Nitrosopiperidine from Tolazamide
Authors: Eshraghi Jamshid , Longo John , Dalton David R. , Harrington George W.
Published: August 2005

The previously reported reaction of tolazamide with nitrite, under physiological conditions, to form N‐nitrosohexamethyleneimine and surprisingly, N‐nitrosopiperidine was confirmed. By using the six‐membered ring analogue of tolazamide, 1‐(piperidyl)‐3‐(p‐tolylsulfonyl)urea, which yields the corresponding N‐nitrosopiperidine and N‐nitrosopyrrolidine, the present study shows that an unusual ring contraction occurs, excising the carbon alpha to the nitrogen.
Induction of Mouse Anti‐melanoma Cytotoxic and Suppressor T Cells in vitro by an Artificial Antigen, GM3‐lactone
Authors: Harada Yoshitada , Sakatsume Minoru , Taniguchi Masaru
Published: August 2005

We investigated the ability of GM3‐lactone liposomes to induce anti‐melanoma T cell responses in mice. GM3‐lactone liposomes, like murine B16 melanoma cells, induced anti‐melanoma cytotoxic T cells (CTL) and also suppressor T cells (Ts). A small dose of GM3‐lactone (0.0003 μg/ml) was enough to generate CTL in the in vitro primary response, whereas relatively large amounts of the antigen (0.03–0.3 μg/ml) were required for anti‐melanoma Ts induction. As the epitope for anti‐melanoma Ts is NeuAc but not NeuGc residue on GM3, and anti‐melanoma CTL are effectively induced by either GM3(NeuAc) or GM3(NeuGc)‐lactone liposomes, GM3(NeuGc)‐lactone or GM3(NeuGc) liposomes have potent activity as an artificial melanoma antigen to induce anti‐melanoma CTL in vitro.
Production of Tumor Necrosis Factor‐α by Alveolar Macrophages of Lung Cancer Patients
Authors: Okubo Akio , Sone Saburo , Singh Sukh Mahendra , Ogura Takeshi
Published: August 2005

The abilities of human alveolar macrophages (AM) obtained from healthy donors and patients with lung cancer to produce tumor necrosis factor (TNF) were compared with those of their blood monocytes after activation with lipopolysaccharide (LPS). TNF activity was assayed by measuring cytotoxicity against actinomycin D‐treated L929 cells and TNF was determined quantitatively by sandwich enzyme‐linked immnnosorbent assay (ELISA) with polyclonal and monoclonal antibodies against TNF‐α. Unstimulated AM from healthy donors released variable amounts of TNF spontaneously, whereas blood monocytes did not. When treated with LPS for 24 h, AM and monocytes produced TNF dose‐dependently, but TNF production by AM was significantly more than that by blood monocytes. This TNF activity was inhibited completely by monoclonal anti‐TNF‐α antibody. Macrophages generated by in vitro maturation of monocytes induced by granulocyte‐macrophage colony‐stimulating factor (GM‐CSF) produced more TNF than freshly isolated monocytes. No difference was found in the abilities of AM from healthy donors and patients with lung cancer to produce TNF after activation stimuli. These observations suggest that human AM may be important in in vivo antitumor defense of the lung through TNF‐α production.
Advantages in Combination Chemotherapy Using Cisplatin and Its Analogues for Human Testicular Tumor Xenografts
Authors: Hida Shuichi , Okada Kenichiro , Yoshida Osamu
Published: August 2005

The antitumor effects and toxicities of combination chemotherapies using cisplatin (CDDP) and its analogues were compared with those of single drug therapies. Congenitally athymic nude BALB/c (nu/nu) mice were used to estimate antitumor activities of these compounds against human testicular tumor (Ht‐14) xenografts and hetero‐BALB/c (nu/+) mice were used to evaluate the toxic effects of the drugs. Combination therapy with half dosages of CDDP and carboplatin (JM8) (CDDP: 2, JM8: 20 mg/kg/day for 5 days), or of CDDP and (glycolato‐O, O′)‐diammineplatinum (II) (254S) (CDDP: 2, 254S: 4 mg/kg/day for 5 days), resulted in significant tumor regression. The combination of CDDP and JM8 had the highest therapeutic efficacy while the CDDP and 254S combination had a lower antitumor potency. In addition, the toxicities of the combination therapies were lower than what was produced by the highest dosage of CDDP (4 mg/kg/day for 5 days). These results demonstrated that the antitumor activities of these combination chemotherapies were equal or superior to the activity of CDDP or an analogue alone, and that the toxicities produced by these combinations were more manageable than those produced by single drug therapies.
Ick Suppresses Gene Expression from Various Promoters Including Human T‐Cell Leukemia Virus Type I Promoter
Authors: Ohta Masataka , Morita Toshirou , Shimotohno Kunitada
Published: August 2005

The T‐lymphocyte‐specific tyrosine kinase gene, Ick, is expressed in T‐lymphocyte cell lines, except for several human T‐cell leukemia virus type I(HTLV‐I)‐transformed T‐lymphocyte cell lines, which produce HTLV‐I. By introducing an Ick‐expression vector, we found that Ick product suppresses gene expression from HTLV‐I promoter in a transient assay. Moreover, various other promoters of cellular genes or viruses were found to have their transcriptional activity repressed by Ic.
Changes in Serum Concentrations of β‐Carotene and Changes in the Dietary Intake Frequency of Green‐Yellow Vegetables among Healthy Male Inhabitants of Japan
Authors: Suzuki Sadao , Sasaki Ryuichiro , Ito Yoshinori , Hamajima Nobuyuki , Shibata Atsuko , Tamakoshi Akiko , Otani Motohiko , Aoki Kunio
Published: August 2005

Serum levels of β‐carotene among 147 healthy male inhabitants were measured twice with an interval of one year in order to determine the relationship between changes in serum β‐carotene levels and changes in the dietary intake of green‐yellow vegetables. A positive association was found to exist between changes in the intake frequency of green‐yellow vegetables and changes in serum β‐carotene levels, whereas changes in alcohol intake and smoking were discovered to be negatively associated with changes in serum β‐carotene levels. The positive association between changes in the intake frequency of green‐yellow vegetables and changes in serum β‐carotene levels was preserved after adjustment for these negative factors.
Two‐dimensional Electrophoretic Analysis of Hepatitis‐associated Polypeptides in Liver of LEC Rats Developing Spontaneous Hepatitis
Authors: Wirth Peter J. , Fujimoto Yoshinori , Takahashi Hidetoshi , Mori Michio , Yoshida Michihiro C. , Sugimura Takashi , Nagao Minako
Published: August 2005

High‐resolution two‐dimensional polyacrylamide gel electrophoresis in combination with silver staining was used to analyze between 800 and 1000 cytosolic and particulate polypeptides from age‐matched livers of normal male Long‐Evans rat with Agouti coat color (LEA) and Long‐Evans rat with Cinnamon‐like coat color (LEC) rats with hereditary trait of hepatitis at ages long before, immediately prior to, and just after the onset of hepatitis. Although the electrophoretic patterns of polypeptide expression were very similar with respect to the overall spot patterns, a number of polypeptides which differed either qualitatively or quantitatively were noted. Two constitutively expressed cytosolic polypeptides, P29.5 (Mr 29.5 kDa/pI 6.73) and P30 (30 kDa/6.70), were not detected in livers of LEC animals at any age. In the normal LEA rats both P29.5 and P30 were detected as early as one day after birth and both were expressed at similar concentrations at all ages. In the LEC rats P30‐C (30 kDa/6.68) was constitutively expressed in close proximity to the expected position of P30, and P30‐C was not detected in the LEA rats. By means of non‐equilibrium pH gradient electrophoresis two relatively basic polypeptides were detected in the LEC rats. P18ne was detected immediately prior to and P27ne immediately after the clinical manifestation of hepatitis. Experiments in F1 backcross ([LEA × LEC] × LEC) animals, however, failed to demonstrate any genetic link between either the expression or lack of expression of P29.5, P30, P30‐C, or P18ne and hepatitis development. P27ne was detected in all backcross animals exhibiting hepatitis, but was never observed in LEC rats prior to the onset of hepatitis. Although we were unable to identify any unique loss of expression of polypeptides which are genetically linked to hepatitis susceptibility in LEC rats, specific subsets of quantitatively modulated polypeptides were detected.
Novel Non‐tumorigenic Cell Variants Showing Potentially Different Susceptibility to v‐src‐induced Metastasis
Authors: Enomoto Taira , Ayaki Hitoshi , Nakamori Shoji , Enomoto Yuko , Inoue Hirokazu , Kakunaga Takeo
Published: August 2005

Two non‐tumorigenic variant cells were isolated from UV‐irradiated Balb/c 3T3 cells on the basis of their different responsiveness in phorbol ester‐induced morphological change (rounding formation). They showed marked differences of lung metastatic potentials after intravenous injections of their v‐src transformants into nude mice; phorbol ester‐resistant variant TR4 cells transformed by v‐src were hypermetastatic, whereas v‐src transformants of phorbol ester‐sensitive variant TR5 cells were not metastatic at all. These different metastatic responses were not observed in v‐K‐ras‐induced transformants of the variants. These non‐tumorigenic variant cells may pre‐acquire the genetic alteration of certain src‐specific and metastasis‐associated factors. This system may be useful for genetic analysis of the induction of metastasis.
Inhibition of Tumor Cell Arrest in Lungs by Antimetastatic Chitin Heparinoid
Authors: Murata Jun , Saiki Ikuo , Matsuno Kazuhiko , Tokura Seiichi , Azuma Ichiro
Published: August 2005

We have investigated the effect of sulfated chitin derivatives on the intravascular events in the metastatic cascade. 6‐O‐Sulfated carboxymethyl chitin (SCM‐chitin III), as well as heparin, significantly inhibited the arrest of B16‐BL6 cells in lungs after co‐injection with radiolabeled tumor cells, but carboxymethylated chitin (CM‐chitin) had no effect. Heparin showed a potent inhibitory effect on tumor cell‐elicited platelet aggregation and on blood coagulation, which can subsequently enhance the survival, arrest and invasiveness of tumor cells, whereas SCM‐chitin III showed much weaker properties. In contrast, SCM‐chitin III was found to inhibit the adhesion of tumor cells to subendothelial matrix, while heparin did not. SCM‐chitin III was still active in inhibiting experimental lung metastasis even in mice which had been pretreated with anti‐asialo GM1 serum or carrageenan to eliminate NK cells or macrophages. Thus, these results suggest that SCM‐chitin‐mediated inhibition of tumor metastases is distinct from that by heparin and may be due to interference with tumor cell arrest in the capillaries and consequently to the inhibition of tumor cell adhesion to subendothelial matrix.
Acquired Expression of hst‐1 in an Autonomous Subline (Chiba Subline 2) Derived from Androgen‐responsive Mouse Mammary Tumor (Shionogi Carcinoma 115)
Authors: Akakura Koichiro , Furuya Yuzo , Sato Naohide , Kodama Takaomi , Teshima Shinichi , Shimosato Yukio , Yoshida Teruhiko , Terada Masaaki , Shimazaki Jun
Published: August 2005

Since growth of Shionogi Carcinoma 115 (SC 115) and its autonomous subline (CS 2) were regulated by fibroblast growth factor‐like peptide, expression of int‐2 and hst‐1 was examined in these cell lines. Hybridization of genomic DNA with long terminal repeat of mouse mammary tumor virus (MMTV) revealed the same pattern of restriction fragments, showing the same integration of MMTV. Although weak expression of int‐2 was noticed in the two cells, clear expression of hst‐1 was seen only in CS 2 cultured with/without testosterone. It is suggested that autonomous growth of androgen‐unresponsive CS 2 is connected with expression of hst‐1.
Changes in Number of α‐Adrenergic Receptor Subtypes in Hepatocytes from Rats Fed 3′‐Methyl‐4‐dimethylaminoazobenzene
Authors: Miyamoto Ken‐ichi , Sanae Fujiko , Kohei Keiko , Nomura Masaaki , Koshiura Ryozo
Published: August 2005

Changes in numbers of α1‐ and α2‐adrenergic receptors in the plasma membranes of hepatocytes from female Donryu rats given feed containing 0.06% of the carcinogen 3′‐methyl‐4‐dimethylamino‐azobenzene (3′‐MeDAB), were examined. α1‐Adrenergic receptors, measured in terms of [3H]‐prazosin binding, decreased to half of the control 2 weeks after the start of this diet, then gradually decreased for the next 22 weeks. α2‐Adrenergic receptors, measured in terms of [3H]clonidine binding, transiently increased 3‐fold over the control at 2 weeks. These changes in the early period of the 3′‐MeDAB diet intake may be related to hepatocarcinogenesis.
Distribution of Human Granulocyte Colony‐stimulating Factor Receptors on Hematopoietic and Nonhematopoietic Tumor Cell Lines
Authors: Kuwaki Tomoaki , Hosoi Takayuki , Hanazono Yutaka , Tsumura Haruhiko , Ishikawa Fuyuki , Miyazono Kohei , Miyagawa Kiyoshi , Takaku Fumimaro
Published: August 2005

Introduction of two tyrosine residues into human granulocyte colony‐stimulating factor (G‐CSF) allowed us to obtain radioiodinated material efficiently. By using this material, specific receptors for human G‐CSF were analyzed. It was found that human G‐CSF receptors were expressed on several myelocytic and monocytic leukemia cell lines. Furthermore, they were also expressed on two chorio‐carcinoma cell lines. However, the human G‐CSF receptors were not detected on other nonhemato‐poietic tumor cell lines examined, indicating that expression of G‐CSF receptors is strictly limited as compared with receptors for granulocyte/macrophage colony‐stimulating factor (GM‐CSF), which are widely distributed on nonhematopoietic tumors.
Correspondence in Cancer History between Husbands and Wives
Authors: Kato Ikuko , Tominaga Suketami , Suzuki Tsuguyoshi
Published: August 2005

To study the effect of the environments shared by spouses on the development of cancer and some chronic diseases, we analyzed the correspondence of disease history in 21,592 fathers and mothers using the baseline data of a population‐based cohort study. The observed number of cases (O) whose parents had the same disease history was statistically significantly greater than the expected (E); the O/E ratio was 1.53 (95% confidence interval (CI): 1.43–1.63) for all malignant neoplasms, 5.22 (95% CI: 2.81–9.70) for esophageal cancer, 1.63 (95% CI: 1.37–1,93) for stomach cancer, 3.01 (95% CI: 1.89–4.79) for colorectal cancer, 3.90 (95% CI: 2.75–5.53) for liver cancer, 3.14 (95% CI: 1.95–5.08) for lung cancer, 6.73 (95% CI: 2.53–17.87) for bladder cancer, 1.66 (95% CI: 1.54–1.78) for apoplexy and 1.67 (95% CI: 1.51–1.86) for heart disease. The results of the present study suggest that the environmental factors shared by family members for a long time may contribute to familial aggregation of cancer and some chronic diseases.
Species and Organ Differences in DNA Adduct Formation and Repair after Treatment with 4‐Hydroxyaminoquinoline 1‐Oxide
Authors: Qin Xiusheng , Nakatsuru Yoko , Tada Mariko , Ishikawa Takatoshi
Published: August 2005

4‐Hydroxyaminoquinoline 1‐oxide (4HAQO) demonstrates obvious organotropic and species specificity in its carcinogenesis and the present investigation concerns 4HAQO DNA adduct formation and repair as studied in various organs of four animal species (rats, mice, guinea pigs and hamsters). Three hours after an iv injection of 10 mg per kg body weight of tritium‐labeled 4HAQO, the major organs were removed and used for assessment of label incorporation in the DNA. The results showed that the DNA binding levels generally correlated well with the reported species and organ specificity of 4HAQO tumorigenesis. For example, rats showed highest DNA binding in the pancreas and kidney, major target organs. The levels of DNA binding in the liver were invariably low in all 4 animal species, in agreement with the lack of hepatocarcinogenicity associated with 4HAQO exposure. A clear relationship between DNA adduct formation and carcinogen dose was also found after treatment of mice with 4HAQO at doses of 1, 5, 10 and 20 mg per kg body weight in all tissues (pancreas, kidney and lung) except for the liver. Comparison of DNA repair processes in rats, a highly susceptible species, and hamsters, a resistant species in terms of 4HAQO carcinogenicity, revealed highest formation and slowest removal of adducts in the target organs of the rat. In the hamster organs and the rat lung and liver, DNA adduct formation was generally low and in the case of elevation in the initial phase, quickly removed.
A Congenic Line of the DDD Mouse Strain, DDD/1‐Mtv‐2/Mtv‐2: Establishment and Mammary Tumorigenesis
Authors: Matsuzawa Akio , Sayama Kazutoshi , Tsubura Airo , Murakami Akira
Published: August 2005

A single dominant gene on chromosome 18, Mtv‐2, controls both the early appearance of mammary tumors and expression of mouse mammary tumor virus (MMTV) in the milk. A congenic DDD mouse strain, DDD/1‐Mtv‐2/Mtv‐2 (DDD‐Mtv‐2), was developed by introducing this gene from GRS/AJms (GR) into DDD/1 mice by repeating 12 backcrosses and subsequent inbreeding using mammary tumors as a marker for selection. Southern blot analysis of the liver DNA from the resulting congenic mice with EcoRI and MMTV‐U3 probe revealed that two DNA fragments corresponding to Mtv‐2 were specifically transferred from GR to congenic mice. Detection of MMTV‐gp52 antigen in the mammary gland and mammary tumor development in DDDfDDD‐Mtv‐2 mice demonstrated the production of infectious mature MMTV by Mtv‐2 in congenic mice. About 80% of breeding DDD‐Mtv‐2 females developed mammary tumors in the course of one‐year follow‐up. The tumor incidence was lower and the tumor age higher than those in GR mice, suggesting less active functioning of the gene on the DDD genetic background. About 70% of these tumors were morphologically classified as pale cell and type P carcinomas peculiar to GR mice. The gene seemed to control the histologic features of mammary tumors. Congenic mice carried an MMTV provirus in an incomplete form on Y chromosome. The DDD‐Mtv‐2 strain will provide a new model for biological and molecular researches into mouse mammary tumorigenesis.