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Recombinant adipogenesis inhibitory factor (AGIF) was purified to homogeneity from the conditioned medium of COS-1 cells transfected with human AGIF cDNA. The amino-terminal sequence analysis of the mature AGIF revealed that AGIF was produced as a precursor consisting of 199 amino acids and processed into a mature form of 178 amino acids by a cleavage between Ala(-1) and Pro(+1). The purified AGIF inhibited the process of adipogenesis in mouse 3T3-L1 preadipocytes, indicating that AGIF directly acts on the cells. AGIF acted as an adipogenic antagonist not only on the extramedullary cell line 3T3-L1 but also on the mouse bone marrow stroma-derived cell line H-1/A, suggesting that this cytokine may regulate adipogenesis in bone marrow.

A cDNA encoding a novel adipogenesis inhibitory factor (AGIF) that inhibits the process of adipogenesis in mouse 3T3-L1 preadipocytes was cloned from a cDNA library of the human bone marrow-derived stromal cell line KM-102. The cloned cDNA contains an open reading frame coding for an AGIF precursor of 199 amino acids. Analysis of the sequence of this cDNA revealed identity of this factor with a recently reported novel cytokine, designated interleukin-11 (IL-11). AGIF/IL-11 may play an important role in stromal cell-associated hematopoeisis through its regulatory action on adipocyte differentiation in the bone marrow microenvironment.

Administration of human hybrid interferon-alpha A/D (IFN) for 5 consecutive days induced antitumor activity in spleen of Meth A fibrosarcoma-bearing mice 9-10 days after cessation of IFN administration. Antitumor activity assayed by the in vivo neutralization test was found in the spleen of responding mice but not in that of progressive mice. This activity was T-cell-dependent and tumor-selective. However, tumor-neutralizing activity was not found in spleen recovered as early as 3 days after cessation of IFN administration when many tumor cells are still surviving in tumor nodules and being attacked by the host. Instead, T-cell-dependent and tumor-selective tumor-neutralizing activity was found in the lymph node of tumor-bearing mice at this early stage. Furthermore, the tumor-neutralizing activity was already detected in the lymph nodes during the course of IFN administration, although there was no difference in the cell composition of the lymph nodes of IFN- and placebo-administered mice. Since IFN-alpha is apparently not a direct modifier of T cells, these results indicate that IFN probably did not directly increase the number of antitumor T cells but instead modified the host-tumor interaction in such a way that, as an early event, the enhanced T-cell response to tumor cells occurred, resulting in an increase in the antitumor T-cell population in lymph nodes. We supposed that, as the tumor regressed, these antitumor T cells would eventually leave the lymph node and migrate to the spleen.

Administration of human interferon-alpha A/D (IFN-alpha A/D) to Meth A fibrosarcoma-bearing mice produced mononuclear blood cells with antiproliferation activity that were able to suppress Meth A growth in vitro. Their production depended on IFN doses and administration intervals. They were short-lived since antiproliferation activity was not detectable in mononuclear cells 2 days after cessation of IFN administration, although it was detectable as long as IFN administration continued. A similar production of antiproliferative mononuclear blood cells by IFN was achieved in tumor-free as well as in tumor-bearing mice, indicating no deleterious effect of the growing tumor on the production of these cells. IFN-induced antiproliferation activity was not tumor-specific since mononuclear blood cells of IFN-administered mice bearing either Meth A and Meth 1, another antigenically distinct fibrosarcoma, suppressed the in vitro growth of both Meth A and Meth 1 cells. T cells were not involved in the in vivo production of antiproliferative mononuclear blood cells by IFN therapy because the cells with this activity were produced in vivo in Meth A-bearing athymic mice and also in Meth A-bearing mice with reduced T-cell populations by the treatment with alpha mouse thymocyte globulin. T cells were not involved in their antiproliferation activity either, since the in vitro T-cell depletion of mononuclear cells did not diminish this antiproliferation activity. Because mononuclear cells surviving the in vitro T cell depletion were positive for AcM.1 (an epitope of activated macrophage) and also because the alpha Mac-1 antibody completely abolished their in vitro antiproliferation activity, this led us to conclude that it was activated monocytes in blood that were produced by IFN in a T-cell-independent fashion and that these monocytes were the cells responsible for the observed in vitro antiproliferation activity.

We examined the modification of host T cells of tumor nodules by interferon (IFN) therapy in mouse models. The host cells were recovered from regressing tumor nodules of mice at Day 13 after intradermal tumor inoculation at Day 0 and administration of 5 x 10(5) U/mouse/day IFN at Day 6 to Day 10. These host cells neutralized in vivo Meth A growth in a dose-dependent fashion. In vitro treatment of these cells with anti-Thy 1.2 monoclonal antibody and rabbit sera as a source of complement abrogated their tumor-neutralizing activity, but only partially, indicating that both T cells and non-T cells were involved in tumor neutralization. The finding that host cells from regressing tumor nodules of either Meth A or Meth 1, an antigenically distinct fibrosarcoma, neutralized both Meth A and Meth 1 tumors without much selectivity was consistent with possible non-T cell involvement. Most of these characteristics of host cells of regressing nodules of IFN-administered mice were also noted with host cells of progressing nodules of placebo-administered mice and there was no significant difference in neutralizing activity qualitatively or quantitatively between the two sources of host cells. There was no significant difference in host T and B cell numbers and compositions of regressing and progressing nodules either. These essentially negative findings raise the possibility, among others, that the primary target host cells to be modified by IFN were not T cells, although the therapeutic effect of IFN was dependent on the host T cells.

Effects of cyclophosphamide (CY) on the antibody titer level and incidence and severity of swine serum (SS)-induced bile duct lesion (BDL) were examined. BDL induced by 0.2 ml of SS per head twice a week for 2 weeks was characterized by hyperplasia of biliary epithelial cells, proliferation of mucous glands, and periductal infiltration of eosinophils with mild fibrosis. CY showed no significant influence on the above-mentioned parameters at the dose levels of 140 and 210 mg/kg. On the other hand, CY lowered the antibody titer level and decreased the severity of BDL at the dose level of 280 mg/kg, and it suppressed the antibody response and BDL at the dose level of 280 x 2 mg/kg. Thus the antibody titer level and the severity of BDL were closely related each other.

Spontaneous glomerular lipidosis was found in a 12-week-old male Syrian hamster of the APA strain. Lipids in the glomeruli were observed as droplets in a prominently expanded mesangial area and as emboli in a dilated capillary lumen. Lipid deposition was also, but less often, detected in tubular epithelial cells and interstitial cells around the lipid-laden glomeruli. This case of glomerular lipidosis was considered to be closely related to hyperglycaemia and hyperlipidaemia.

Safracins A and B have antibacterial activity against Gram-positive and Gram-negative bacteria in vitro but no therapeutic activity in mice infected with Staphylococcus aureus. Safracins A and B induce abnormal morphological changes in Echerichia coli cells. Tests with transplantable mice tumors demonstrate that safracins A and B inhibit the growth of P388 leukemia and IMC carcinoma.

Histopathological examinations were carried out on female DBA/2N and CD-1 mice which were autopsied 4 and 12 weeks after six daily intraperitoneal injections of streptozotocin (SZ). Histopathological changes related to SZ treatment were found in the pancreas, liver and kidneys. Little difference was observed between the two strains in the histological changes of the pancreas (a decrease in size of the islets, and degranulation and a decrease in the number of B cells) and liver (hypertrophy of hepatocytes and cytoplasmic invagination into hepatocyte nuclei). With regard to the changes in the kidneys, DBA/2N mice showed characteristic inclusions positive to periodic acid-Schiff reagent in the distal tubule epithelial cells, while CD-1 mice showed remarkable luminal dilatation and epithelial cell deformation of distal tubules. SZ-induced diabetes had no influence on the development of spontaneous cardiovascular lesions in DBA/2N mice under the present experimental conditions.

Calcified tongue lesions were observed in 4- to 8-week-old DBA/2NCrj mice, the highest frequency being reached at 6 weeks of age. Calcification was initially observed in the superficial longitudinal muscles and these lesions were distributed segmentally under the dorsolateral mucous epithelium in the middle portion of the lingual body. After calcification, marked inflammation was observed in the same lesion. Finally, the swollen tissue elevated the covering mucous epithelium and produced polypoid lesions.

A comparative study was conducted of the urinary level of gamma-carboxyglutamic acid (gamma-Gla) in patients with infectious diseases to investigate the mechanism of hemorrhagic diathesis occurrence resulting in post-administrative side effects of cephem-group antibiotics. The results showed that the group of administered antibiotics with N-methyl thiotetrazole (NMTT) side chains exhibited decreases in the urinary level of gamma-Gla. Especially, in the aged group of patients (over 65) with non-carcinoma infectious diseases, significant decreases in the urinary level of gamma-Gla were observed. From the above, it is considered that the administration of antibiotics with NMTT side chains has an effect of prohibiting the vitamin K function.

Intraperitoneal inoculation of L1210 murine leukemia cell vaccine increased I-Ad antigen-positive and -negative macrophages in the peritoneal cavity of histocompatible mice. Mitomycin C, a poor producer of I-Ad antigen-positive macrophages, selectively augmented the production of I-Ad antigen-positive macrophages in L1210 vaccine-primed mice, since it did not augment the production of non-macrophage cells. Since our previous study showed that the priming of mice with L1210 vaccine and mitomycin C induced augmented antitumor response, we tested the feasibility of the association of mitomycin C-augmented I-Ad antigen-positive macrophages with the augmented antitumor response. Prostaglandin E2 suppressed the antitumor response in L1210 vaccine- and mitomycin C-primed mice and, consistent with this, it inhibited the production of I-Ad antigen-positive macrophages, although it inhibited the production of non-macrophage cells as well. This hypothesis was further tested by the use of silica. When L1210 vaccine- and mitomycin C-primed mice were further given silica on the day of and one day after live L1210 inoculation, their antitumor response was strongly suppressed, while a kinetic analysis of the cellularity of peritoneal cells showed that administration of silica resulted in a decrease of I-Ad antigen-positive macrophages, but not I-Ad antigen-negative macrophages or non-macrophage cells in these mice. These results strongly suggest, though they do not prove, the association of mitomycin C-augmented production of I-Ad antigen-positive macrophages with the antitumor response in L1210 vaccine- and mitomycin C-primed mice.

A sensitive method is described for detecting circulating carcinoembryonic antigen (CEA) by combined use of counter-immunoelectrophoresis and intensifiers. Perchloric acid extraction of serum was followed by counter-immunoelectrophoresis against monospecific antiserum to CEA. A precipitin line formed was intensified further by electrophoretic binding of anti-gammaG gamma-globulin and Amido Black staining. Less than 10 ng CEA/ml was detectable. Using this method, a total of 84 sera from patients with or without malignancy was examined. The results obtained were in general agreement with those reported by radioimmunoassay. The method appears to be useful as a diagnostic aid, since it requires neither labeled preparations of purified CEA nor special equipment for assay.

The terminal oxygenase of the NADH-depending lathosterol (cholest-7-en-3 beta-ol) 5-desaturase system was partially purified from rat liver microsomes, by Triton X-100 solubilization, DEAE-cellulose column chromatography, and hydrophobic affinity chromatography with Aminohexyl-Sepharose. The terminal oxygenase activity was approximately 18 fold greater than the starting microsome, and the yield was 18.4%, nevertheless, the terminal enzyme activity was almost free from other electron transfer components in microsomes. It was demonstrated that NADH, molecular oxygen, phospholipid, and three enzymes: NADH-cytochrome b5 reductase, cytochrome b5, and the terminal oxygenase, were absolutely essential for lathosterol 5-desaturation in the reconstituted system. Furthermore, the rate of the NADH-depending lathosterol 5-desaturation in the reconstitution system, was proportional to the concentration either of the terminal desaturase, cytochrome b5, or NADH-cytochrome b5 reductase, under conditions in which other enzymes were present in excess.

A 54-year-old male complaining mainly of pain in the right terminal rib area was admitted to hospital for hepatic tumor examination. Cholangiocarcinoma was diagnosed by various imaging techniques, and the patient was started on a program of daily oral administrations of 600 mg of UFT after abdominal arterial injection of 30 mmg of MMC. The tumor continued to decrease in size on CT image during two years of UFT administration. The patient continues to be monitored as an outpatient. A marked reduction in the size of a cholangiocarcinoma was observed in the case, and administration of UFT has proved effective in prolonging life for over two years.

Electron microscopic observations were carried out on the biliary epithelial cells of BALB/c and DDY mice which had received an intraperitoneal injection of 0.2 ml of swine serum twice a week for 2 or 4 weeks. The most characteristic feature of the biliary epithelial cells of BALB/c mice was a marked increase in the number of vesicles having a close spatial relationship with the well-developed Golgi apparatus or rough endoplasmic reticulum (rER). In contrast, marked dilation of rER filled with moderately electron-dense material was conspicuous in the biliary epithelial cells of DDY mice. A prominent increase in the number of blebs and lateral and basal cytoplasmic protrusions in the dilated intercellular space of the biliary epithelium, and submucosal eosinophil infiltration, collagen fiber proliferation and gland hyperplasia with increased mucin secretion were common to both strains. This experimental model of bile duct disease also seems to be useful for investigating alteration of protein synthesis and secretion in epithelial cells.

The bile duct system of BALB/c and DDY mice, which were immunized with swine serum (SS) or not, was examined histochemically. Biliary epithelial cells of the SS-treated BALB/c mice, which were positively stained with periodic acid-Schiff (PAS) and had binding sites of Dolichos biflorus (DBA), were thought to secrete neutral glycoproteins with terminal N-acetyl-D-galactosamine residues. Those of the SS-treated DDY mice were however negatively or weakly stained with any histochemical stainings. On the other hand, glandular epithelial cells of the SS-treated mice of both strains, which were positively stained with high iron diamine-alcian blue (HID-AB) and had binding sites of DBA, Griffonia simplicifolia-II (GS-II), Ulex europaeus-I (UEA-I), and Triticum vulgaris (WGA), were thought to secrete glycoproteins with terminal sialic acid residues. Biliary and glandular epithelial cells of the normal mice contained only a small amount of glycoproteins showing similar histochemical characteristics to those in the SS-treated BALB/c mice. BALB/c mice immunized with SS were thought to be very useful for the investigation of production and secretion of glycoproteins in the bile duct system as well as being good model of bile duct disease.

The terminal oxidase of the NADH-dependent lathosterol 5-desaturation system was solubilized from rat liver microsomes with 2% Triton X-100, and partially purified approximately 18-fold with 19% yield after DEAE-cellulose and 6-aminohexyl-Sepharose column chromatography. The final enzyme preparation was free from other electron transfer components and phospholipids in microsomes, and the desaturation reaction was reconstituted with the following components: NADH, molecular oxygen, phospholipids and three proteins, i.e., NADH-cytochrome b5 reductase, cytochrome b5 and the terminal oxidase. Omission of one of these components led to an almost complete loss of the desaturase activity. Under the reconstitution conditions, the desaturase activity was significantly inhibited by potassium cyanide but was not affected by -SH reagents such as N-ethylmaleimide and dithiothreitol.

The detailed histopathology of the swine-serum (SS)-induced bile duct lesion and some factors influencing the incidence and severity of the lesion were examined. The lesion was expressed as an eosinophilic and proliferative cholangitis affecting almost the full length of the biliary tract. It appears to be a useful new model of bile duct diseases. The co-existence of sufficient anti-SS antibody together with repeated injection of SS were necessary to produce this lesion. The lesion could be induced reproducibly by intraperitoneal injection of 0.05 to 0.2 ml of swine serum twice a week for 4 weeks in young BALB/c mice.

With virus-infected autologous and allogenic mononuclear cells as specific targets, the development of cell-mediated cytotoxic reactivity to respiratory syncytial virus (RSV) was studied in peripheral blood lymphocytes (PBL) in groups of infants with acute RSV infection and in other control groups of subjects during a community outbreak of RSV infection. No RSV-specific cellular cytotoxicity was observed in cord blood lymphocytes and in other uninfected controls. The PBL of infants with acute RSV infection exhibited significant cellular cytotoxic response. The activity peaked early, usually within 1 week after infection. The response appeared to be age-dependent. Over 65% of infants 6-24 months of age and about 35-38% of infants under 5 months of age exhibited cellular cytotoxicity to RSV. Cellular cytotoxic reactivity was observed against autologous and less frequently against allogenic RSV-infected target cells. These findings suggest the appearance of virus-specific cell-mediated cytotoxic immune response after acute RSV infection. The development of cellular cytotoxic responses may play a role in the mechanisms of protection against or the pathogenesis of RSV infection in man.

The mouse strain difference in bile duct lesions was studied on male A/J, BALB/c, C57BL/6, C3H/He, DBA/2 and DDY mice 4 weeks old given intraperitoneal injections of swine serum (0.05 or 0.2 ml per mouse) twice a week for 4 weeks. The hepatic lesions were restricted to the portal tract. Biliary epithelial cells showed hypertrophy and hyperplasia, and eosinophilic and homogeneous or needle-shaped material appeared in the cytoplasm of such hypertrophied epithelial cells and in the ductular lumen. Around these damaged biliary epithelia, eosinophil leukocyte and plasma cell infiltration with proliferation of collagen fibres was commonly detected. These changes became more apparent with increasing size of bile duct. Such histopathological characteristics of hepatic lesions were essentially the same in all strains, but the severity showed a clear strain difference: the lesion was marked in the DDY, A/J and BALB/c strains, moderate in C3H/He and slight in C57BL/6 and DBA/2. A high production of anti-swine-serum antibodies associated with a marked increase in the number of mouse IgG-producing lymphocytes in the spleen was detected in the strains showing the marked hepatic lesions.