Mosaicism in the expression of tumor-associated carbohydrate antigens in human colonic and gastric cancers.

Serial sequential sections from a single tumor were examined by immunohistological staining with several monoclonal antibodies directed, respectively, to different tumor-associated carbohydrate epitopes. Staining patterns were compared with those of conventional staining with hematoxylin-eosin or periodate/Schiff's reagent. Each tumor showed different areas of staining with different antibodies, and the combined staining map shows a clear mosaicism of antigen expression within the same tumor. For example, some areas of a given tumor were stained by FH4 (defining dimeric Le(x)), while other complementary areas were strongly stained, in a mutually exclusive manner, by SH1 (defining Le(x)), AH6 (defining Le(y)), FH6 (defining sialosyl dimeric (Le(x)), or TKH2 (defining sialosyl-Tn). Some areas were stained by two or three of these antibodies. Comparisons of the mosaic-staining patterns with cytohistological properties of tumor cells within specific areas suggested that the pattern of antigen expression is correlated with degree of differentiation; e.g., poorly-differentiated cells with severe dysplasia did not express high levels of Le(x) or Le(y) but did express sialyl-Le(x) or dimeric Le(x); on the other hand, moderately or well-differentiated tumor cells in some areas expressed high levels of Le(x) or Le(y) but lower levels of sialyl-Le(x). Areas showing strong expression of sialyl-Tn in their secretions were consistently correlated with presence of well-differentiated tumor cells, whereas secretions from normal mucosae were consistently characterized by lack of sialyl-Tn expression. It is postulated that the original in situ tumors (which had homogeneous glycosylation patterns) evolved into several spatially discrete cell populations displaying different degrees of glycosylation, reflecting stages of tumor cell differentiation and progression.

Effects of oral lentinan on T-cell subsets in peripheral venous blood.

The effect of oral lentinan, a biological response modifier, on the control of systemic immune function was studied in six-week-old male Wistar-Imamichi specific-pathogen free rats. In the lentinan-treated group, 1 mg of lentinan dissolved in 1 ml of physiological saline was administered forcibly into the stomach twice weekly for four or eight weeks. Physiological saline alone was administered in a similar fashion to the control group. Leukocyte and lymphocyte counts were made and lymphocyte subsets measured using monoclonal antibodies W3/13, W3/25, and OX8, and a laser flow cytometry system. The T-cell level, the helper/inducer T-cell level, and the suppressor/cytotoxic T-cell level were measured. The peripheral leukocyte and lymphocyte counts did not change significantly in either group during treatment. After four weeks of treatment, however, the lentinan group had a significantly higher T-cell level, helper-cell level, and helper-suppressor ratio, and a significantly lower suppressor-cell level than did the control group. No significant between-group differences in the lymphocyte subsets or the helper-suppressor ratio were noted after eight weeks of treatment. Oral administration of lentinan appears to modulate the systemic immune function through stimulation of T cells, especially helper cells. Continued administration produced less effect, possibly due to a tolerance to the effect of lentinan.

Postoperative intravenous administration of lysine acetylsalicylate: effect on pain relief and platelet function.

The effects of lysine acetylsalicylate, a new injectable salicylate, on postoperative pain relief and platelet function were studied in ten men who had surgery for peptic ulcer. Four of five patients receiving lysine acetylsalicylate had satisfactory pain relief. One patient required an additional injection of 15 mg of pentazocine. Of the five patients in the control group, an average (+/- SD) dose of 90 +/- 23.7 mg of pentazocine was required to achieve adequate postoperative pain relief. Lysine acetylsalicylate decreased platelet aggregability but without resulting in hemorrhage. We concluded that this new salicylate administered intravenously to patients in the postoperative period provided adequate analgesia while allowing effective hemostasis despite its inhibitory effect on platelet aggregation.

The effect of a protein-bound polysaccharide on T cell subsets of peripheral venous blood and thoracic duct lymph.

To investigate the effect of a protein-polysaccharide preparation, PSK, on immune system function, the ratios of peripheral venous blood and thoracic duct lymphocyte subsets were determined in 6-week-old male SPF Wistar-Imamichi strain rats maintained on a diet containing 2% PSK ad libitum. Peripheral blood and thoracic duct lymph specimens were obtained from pretreatment, 4- and 8-week PSK-treated, and period-matched control groups of rats. The lymphocyte count and percentages of helper/inducer and suppressor/cytotoxic T cells were determined with a laser flow-cytometry system using monoclonal antibodies W3/13, W3/25 and OX8. Compared with the controls, the PSK-treated group showed significantly lower total T cell and helper/inducer T cell counts in the peripheral blood, but showed no difference in the suppressor/cytotoxic T cell count. However, in the thoracic duct, there were significantly increased total T cell and helper/inducer T cell counts, but no difference in suppressor/cytotoxic T cell counts compared with the controls. These data appear to indicate a significant activity of oral PSK, resulting in modulation of lymphocyte distribution in the body by facilitation of the transition of circulating T lymphocytes, especially helper/inducer T cells, into the lymph.

Inhibitory effect of simultaneous intraportal administration of 5-fluorouracil, uracil and degradable starch microspheres on experimental hepatic micrometastasis, is of colon cancer.

This study was designed to examine the optimal regimen of 5-fluorouracil (5-FU), uracil and degradable starch microspheres (DSM) to prevent the hepatic metastasis of colorectal cancer. BALB/C mice were used as a model of hepatic micrometastasis of mouse transplantable colorectal cancer, Colon 26. We intraportally administered to the mice 5-FU at a dose of 34.8 mg/kg body weight; uracil at the dose equal to, or 8 times higher than that of 5-FU in molar weight and DMS at a dose of 5 mg/kg body weight. We investigated the hepatic and serum concentrations of 5-FU and uracil and the inhibitory effect of various combinations of the three substances on hepatic metastasis. The study showed that concurrent use of 5-FU with uracil at a dose 8 times higher than that of 5-Fu could keep 5-FU concentration high in hepatic tissue and relatively low in serum. This tendency became more marked with the presence of DSM. These findings support the previous study results that uracil antagonizes breakdown of 5-FU in the liver and with the aid of DSM, retards excretion of 5-FU from the liver. Further investigations for clinical application of this method will lower the incidence of hepatic metastasis of colorectal cancer.

Effect of the proteoglycan (PSK) on lymphocyte subsets in normal rats.

The effect of the proteoglycan biologic response modifier, PSK, on lymphocyte subsets was investigated in normal rats. Six-week-old male SPF Wistar-Imamichi rats were fed a diet containing 2% PSK. Peripheral blood lymphocytes (PBL), thoracic duct lymphocytes (TDL), and those existing in the thymus, spleen and Peyer's patches were collected for analysis of subsets of T cells, helper/inducer T (Th) cells, suppressor/cytotoxic T (Ts) cells and B cells compared with those of the control group. In the PBL, differential and absolute counts of T and Th cells were lower in the group fed PSK (the PSK group) than the control group. There was no difference in Ts cells between the groups, and the PSK group showed a higher B-cell differential count. In the case of TDL, the PSK group showed greater absolute counts of T and Th cells than the control group. In tissue lymphocytes, differential T and Th cell counts were significantly greater in the PSK group than the control group, as observed first in Peyer's patches and later in the spleen. No differences between the groups were observed in these counts in the thymus. Changes in body distribution of T and Th cells induced by PSK treatment first appeared in lymphocytes in Peyer's patches, followed by PBL and TDL, and those in the spleen. No such changes were observed in the thymic lymphocytes.

Two cases of adenolipoma of the breast.

Adenolipoma of the breast is a rare tumor. Two cases are described here and the literature is reviewed. Case 1 was a 52-year-old woman who had a 3-cm-long elastic hard tumor in the left breast. Case 2 was a 40-year-old woman who had a 3-cm-long elastic soft tumor in the left breast. In each case, mammography revealed well demarcated soft tissue density. Ultrasonography demonstrated well defined tumors composed of echogenic and sonolucent areas. These tumors were easily enucleated at surgery. Histologically, the tumors consisted of mammary ducts, fibrous stroma and adipose tissue, findings which are characteristic of mammary adenolipoma. An adenolipoma may still be misdiagnosed as a fibroadenoma or a fibrocystic disease. Characteristic findings of mammography and ultrasonography are emphasized.

Hemolytic streptococcus preparation OK-432; beneficial adjuvant therapy in recurrent gastric carcinoma.

The administration of a hemolytic streptococcus preparation, OK-432, is thought to be of therapeutic value in the treatment of cancer patients through a stimulatory effect on the immune system. In order to evaluate any beneficial effect of such an administration, a group of patients with recurrent gastric cancer was studied. This group was randomly subdivided into 3 groups: 1) Intradermal group (ID Group), 42 patients given an intradermal injections of OK-432. 2) Intramuscular group (IM Group), 40 patients given an intramuscular injections of OK-432. 3) Control group (C Group), 39 patients not given injections. Each group was studied in regards to the length of survival, the host immune response and the incidence of side effects. When compared to the IM and C Groups, the ID Group showed improved survival. Accompanying this improved survival, the ID Group also had greater white cell and lymphocyte counts, a greater number of T cells, and a more dramatic skin reaction to the extracted cell wall polysaccharide of hemolytic streptococcus Su-strain (Su-PS). The ID Group, following OK-432 injection, had a 4.8% incidence of fever and a 52.4% incidence of local abscess formation at the injection site. 90% of the IM Group developed fever but abscess formation was absent in all patients. From these results, it was concluded that in the patients tested, intradermal injection of OK-432 appears to be clinically superior to intramuscular injection.

Effects of biological response modifier on thoracic duct lymphocytes in recurrent gastric cancer. Evaluation of OK-432, a hemolytic streptococcus preparation.

In 9 patients with recurrent gastric carcinoma treated with intracutaneous injections of a hemolytic streptococcal preparation of OK-432 (PIC), the T/B cell ratio, the number of IgG-Fc receptor positive (T gamma) cells, the degree of blastoid transformation of lymphocytes in response to concanavalin-A (Con-A) and phytohemagglutinin (PHA), and the activity of natural killer cells (NK) in the thoracic duct lymphocytes (TDL) and peripheral blood lymphocytes (PBL) were measured. In thoracic duct lymph, the T cell to B cell ratio was higher than that found in peripheral blood. The proportion of T gamma cells and the natural killer cell activity was found to be considerably lower in TDL than PBL, though there was a higher degree of blastoid transformation of lymphocytes in response to Con-A and PHA in TDL. After the administration of PIC, there was no significant difference in T/B cell ratios and T gamma cell proportion between TDL and PBL, though the lymphocytic blastoid transformation in response to Con-A and PHA decreased in TDL but not in PBL. PIC administration appeared to augument natural killer cell activity in both TDL and PBL.

Effects of oral administration of streptococcal preparation OK-432 on thoracic duct lymphocytes.

One KE of streptococcal preparation OK-432 (PIC) was given orally to 6-week old male Wistar-Imamichi rats twice weekly, and control animals received comparable volumes of saline alone. Samples of thoracic duct lymph and posterior vena caval blood were collected prior to and 4 and 8 weeks following PIC. The T cell count of thoracic duct lymphocytes was about three times greater than that of venous blood specimens, and increased significantly in animals following PIC. Controls showed only a slight increase followed by a decrease, so that by 8 weeks of treatment with PIC, the thoracic duct T cell count revealed a significantly greater number in the PIC group than in the controls. In addition, after 8 weeks the per cent T cell subpopulation in thoracic duct lymph increased significantly in the treated group, but it declined in the control group. Concerning T cell subsets in thoracic duct lymphocytes, the percentage of suppressor/cytotoxic T cells showed a significant elevation in the control group after 4 weeks compared with the treated group. No other blood or lymph variables reached statistical significance in this respect.

Estimation of the functional reserve of the human liver by urinary D-glucaric acid excretion after vitamin C administration.

The excretion of D-glucaric acid in the urine (uGA) correlates with the total liver content of hepatic cytochrome P-450, the metabolism of which depends on adenosine triphosphate (ATP) being produced by intrahepatic cellular mitochondria. Five cases of compensated liver cirrhosis group with less than 0.4 mg/kg/min of the maximum removal rate of indocyanine green (ICGR max), and 5 cases with normal hepatic function (control group), were monitored for uGA before and after P-450 activation induced by administration of 1 g of vitamin-C. Before vitamin-C administration, no differences in uGA excretion were observed comparing the cirrhosis with the control group. After administration of vitamin-C, the excretion of uGA was significantly lower in the cirrhosis group. The measurement of uGA is considered to represent vitamin-C induced activation by P-450, and is a new method for evaluation of the functional reserve of the liver.

A case of microhepatoma associated with schistosomiasis japonica diagnosed by enhanced ultrasonography after hepatic intraarterial injection of carbon dioxide gas.

The patient was a 58-year old man whose complaints were generalized malaise and right epigastralgia. He had liver cirrhosis and schistosomiasis japonica, previously diagnosed by laparoscopy. Computed tomography (CT) showed a high density funicular shadow in the liver. However no tumorous lesions in the liver were visualized. Ultrasonography (US) of the liver showed a reticulate or scaly pattern, but no images of tumorous lesions. Hepatic angiography showed a single, deeply colored image about 1cm in diameter, in the segmentum anterosuperior. Preoperative and intraoperative enhanced US with hepatic intraarterial injection of carbon dioxide gas was performed. It showed a hyperechoic tumor shadow about 1cm in the segmentum anterior. The segmentum anterosuperior including the tumor was partially resected. Pathologically, the tumor was found to be a hepatoma of Edmondson type II, caused by cirrhosis and schistosomiasis japonica. The patient's postoperative course was uneventful. Enhanced US with hepatic intraarterial injection of carbon dioxide gas was useful for the diagnosis and treatment of the microhepatoma associated with schistosomiasis japonica.

[The influence of a biological response modifier on thoracic lymphocytes--a study involving clinical cases and animal experiments].

The influence of a biological response modifier on thoracic lymphocytes was studied clinically and experimentally. The thoracic lymphocytes and peripheral blood were taken from nine cases of gastric cancer relapse before and after the administration of hemolytic streptococci, OK-432 (PIC), for immunological examination. T: B cell ratio, T cell ratio and ADCC activity of the thoracic lymphocytes and peripheral blood were not changed but the NK activities of both were increased. The juvenilization of lymphocytes was reduced but that of the peripheral blood was not changed. The number and the ratio of T cells in the thoracic lymphocytes of Wistar rats were considerably increased by the administration of PIC but they were reduced in the peripheral blood. T-cell subsets of thoracic lymphocytes and peripheral blood were hardly changed by the administration of PIC. In the case immunological treatment, the thoracic lymphocytes and blood lymphocytes should be monitored because their biological specificities are different.

[An immunological study on the effect of simultaneous intrahepatic injection of an immunomodulator (OK-432) and adriamycin--an experiment in rats].

An immunological study of the effect of a simultaneous intrahepatic injection of adriamycin (ADM) and OK-432 (PIC) in seven-week-old S.D. rats was carried out. Three experimental groups included an ADM-PIC-0.5 group which was administered 0.5 mg/kg ADM and 0.5 KE/kg PIC, an ADM-PIC-1.0 group which was administered 1.0 mg/kg ADM and 1.0 KE/kg PIC and an ADM-PIC-2.0 group which was administered 2.0 mg/kg ADM and 2.0 KE/kg PIC. A control group was injected with normal saline. The numbers of T-cells, B-cells, helper T-cells and suppressor T-cells were counted before, 1 day after and 5 days after intrahepatic injection. The suppressor T-cell count decreased (p less than 0.05) in the ADM-PIC-0.5 group and the T-cell count (p less than 0.05), B-cell count (p less than 0.05), helper T-cell count (p less than 0.01), suppressor T-cell (p less than 0.05) and the ratio of suppressor T-cells among lymphocytes were decreased in the ADM-PIC-1.0 group. Decreases in the T-cell count (p less than 0.01), B-cell count (p less than 0.05), helper T-cell count (p less than 0.01), suppressor T-cell (p less than 0.01) and the ratio of suppressor T-cells among lymphocytes (p less than 0.01) were recognized in the ADM-PIC-2.0 group one day after injection. Five days after injection, the ratio of suppressor T-cells among lymphocytes (p less than 0.01) was decreased in the ADM-PIC-0.5 group, and suppressor T-cells (p less than 0.05) and the ratio of suppressor T-cells among lymphocytes were decreased in the ADM-PIC-1.0 group. In the ADM-PIC-2.0 group, suppressor T-cell count (p less than 0.01) and the ratio of suppressor T-cells among lymphocytes were decreased. The result suggested that a simultaneous intrahepatic injection with PIC could improve the immunosuppression produced by an intrahepatic injection of ADM.

[Prospective randomized trial comparing modified FAM (5-fluorouracil (5-FU) + adriamycin + mitomycin C) versus 5-FU alone for the treatment of non-resectable pancreatic and biliary tract carcinomas (the 1st trial in non-resectable patients). Study Group of Surgical Adjuvant Therapy for Carcinomas of the Pancreas and Biliary Tract].

The modified FAM (5-fluorouracil (5-FU) + adriamycin (ADR) + mitomycin C (MMC)) therapy (FAM group) was compared with 5-FU mono-therapy (F group) by multi-institutional randomized trial in the patients with cancer of the pancreas or the biliary tract who underwent non-resection. The patients in FAM group received 6 mg/m2 of i.v. MMC during operation, 310 mg/m2 of i.v. 5-FU for 5 days in the 1st and 3rd postoperative weeks and 12 mg/m2 of i.v. ADR in the 2nd postoperative week. Those in F group received only 5-FU course in the administration schedule of FAM group. Among the cases which completed respective whole administration schedules. 35 cases in FAM group and 36 in F group, better effect than partial response (PR) was observed in neither groups, and there was no significant difference between groups with respect to overall/each disease survival duration, progression-suppressed duration and clinical effect. Primary adverse effects were alimentary symptoms and hepatic dysfunction, neither of which was serious, and there was no difference between groups except that hair loss was observed in more cases in FAM group (p less than 0.05). Results in FAM group did not statistically surpass those in F group, but a tendency was observed that FAM group was better than F group in terms of survival duration and clinical effect for cancer of the gall-bladder.

[Immunohistochemical studies on tumor associated carbohydrate antigens in gastric cancers of different histological stages].

By comparing gastric cancer tissues treated by the conventional hematoxylin and eosin (HE) staining and those by the chemical staining of immunohistologicals using monoclonal antibodies (MoAB) which recognize different carbohydrate antigens, the relation of cancer tissue patterns between the two staining methods was studied. Consequently, stainability was not seen in MoAB-FH4, AH6, FH6 and TKH2 in the cancer tissues where MoAB-SH1 responded to immunohistological staining. Likewise, each of MoAB-FH4, AH6, FH6 was found to have its own stain localization. The patterns made by immunohistological staining using MoAB showed so-called mosaicism even where the HE stain presented the same histologic form. Study of correlation between gastric cancer patterns and MoAB's localization revealed that localization of MoAB-SH1, AH6 and TKH2 was predominant in well differentiated adenocarcinoma. On the contrary, MoAB-FH4 and FH6, which are more specific, showed predominant localization in poorly differentiated adenocarcinoma of gastric cancers. Localization of MoAB-FH6 and AH6 increased as cancer grew from the early stage to the advanced stage. These results leads to this assumption: cancer, being of an isogenic carbohydrate structure at the initial stage when carcinoma in situ is generated, gains heterogeneity with the process of growth, differentiating into various directions and thus changing into a complicated carbohydrate structure.