Search

Antibody-dependent macrophage-mediated cytotoxicity was studied to determine the significance of cytophilic anti-thyroglobulin antibody (ATgA) present in the sera of patients with Hashimoto's thyroiditis. Effector cells were normal human monocytes or guinea-pig peritoneal exudate cells, and target cells were human thyroglobulin(Tg)-coated chicken erythrocytes. Cytotoxicity was evaluated by morphological observation and by 51Cr-releasing assay. Normal human monocytes rapidly destroyed ATgA-bound Tg-coated chicken erythrocytes by extracellular cytolysis and by phagocytosis. On the contrary, human monocytes "armed" with cytophilic ATgA destroyed Tg-coated chicken erythrocytes slowly and to a lesser extent, and only by extracellular cytolysis. When normal monocytes or peritoneal exudate cells were incubated with Tg-coated chicken erythrocytes in the presence of the sera of patients with Hashimoto's thyroiditis, phagocytosis occurred rapidly, but extracellular cytolysis developed rather slowly. These data suggest the possibility that human monocytes participate in antibody-dependent cell-mediated cytotoxicity (ADCC) in vivo, which may be an important destructive mechanism in Hashimoto's thyroiditis. It is also possible that ATgA cytophilic for monocytes render non-immune peripheral monocytes cytotoxic against Tg-bearing cells.

To investigate factors involved in excision repair DNA synthesis, a soluble extract was prepared from permeable mouse sarcoma (SR-C3H/He) cells by homogenization and ultracentrifugation. DNA synthesis measured by using native calf thymus DNA as the template-primer and the extract as the polymerase source showed low activity. The DNA synthesis was enhanced more than ten-fold by the addition of an appropriate concentration of bleomycin, a radiomimetic DNA-damaging drug. Using selective inhibitors of DNA polymerases, it was shown that the DNA polymerase involved in the bleomycin-induced DNA synthesis was DNA polymerase beta. In addition to DNA polymerase beta, an exonuclease which converts bleomycin-damaged DNA into suitable template-primers for repair DNA synthesis appeared to be present in the permeable cell extract.

The difference in the physiological condition of drivers of manual transmission buses (MTB) and automatic transmission buses (ATB) was examined from the viewpoint of occupational health. This study was based on a self-administered questionnaire which involved items concerning subjective fatigue complaints. No differences in the mental fatigue and stress between MTB drivers and ATB drivers were observed. Although ATB drivers tended to feel less physical fatigue than MTB drivers, the difference was not statistically significant. From these results, it was suggested that there was little difference in the subjective fatigue between ATB drivers and MTB drivers.

The analysis of bile acids in human bile was tried by high-performance liquid chromatography (HPLC). Bile acids in human bile were first prefractionated into free, glycine- and taurine-conjugated bile acids using a Seppak C18 cartridge and a piperidinohydroxypropyl Sephadex LH-20 (PHP-LH-20) column. Each fraction was then processed through a HPLC system consisting of a Zorbax ODS column and a 3 alpha-hydroxysteroid dehydrogenase (3 alpha-HSD) column. By these procedures the major 15 bile acids were clearly separated, and each bile acid of 10-125 ng was accurately analyzed. More than 400 times of analysis could be repeated on one 3 alpha-HSD column without loss of sensitivity. Thus the pretreatment through Seppak C18 and PHP-LH-20 made the HPLC analysis of human bile acids accurate and applicable to clinically obtained materials.

We administered serum fractions obtained from cancer patients by double-filtration plasmapheresis (DFPP) to cancer-bearing mice to examine the effects on tumor growth and metastasis. Fraction 1 (whole plasma), fraction 2 (a plasma fraction containing substances with higher particle size), fraction 3 (a plasma fraction containing substances with smaller particle size) and saline were administered intravenously to cancer-bearing mice for 10 days following the inoculation of tumor cells. The tumor growth and metastasis in mice administered fraction 2 was far more rapid than that in the control mice. On the other hand, tumor growth in mice administered fraction 3 was significantly delayed compared with that in mice injected with fraction 2. These results suggest that factors in the higher particle-size fraction of cancer patients' sera promote the growth and the metastasis of tumors in mice, and that DFPP, which remove these factors, is an effective therapy against cancer.

A clinical trial was performed to investigate the efficacy of hyperthermia in combination with chemotherapy for gynecological malignancies. Sixty-nine patients with vaginal or vulvar malignancies (9 primary vulvar, 3 recurrent vulvar, 11 primary vaginal, 4 primary cervical, 40 recurrent cervical, and 2 recurrent ovarian carcinomas) were treated by thermochemotherapy (42 cases) or chemotherapy alone (27 cases). After treatment, 7 patients underwent surgery and 46 patients irradiation. The chemotherapeutic schedule was mainly a combination therapy with bleomycin and mitomycin C (B-M). Microwaves of 2.45 GHz were applied to induce local hyperthermia. The side effects of chemotherapy were not increased by hyperthermia. The rate of partial response plus complete response increased to 84% (16/19) in primary cancers and 45% in recurrent cancers by hyperthermia, compared to the respective values of 40% (2/5) and 17% (3/17) for chemotherapy alone. However, a satisfactory prognosis cannot be expected with thermochemotherapy, unless additional treatments are performed. Subsequent surgery or radiation treatment improved the progression-free interval.

A significant amount of anticoagulant substance was released along with histamine, when human lung mast cells were stimulated with anti-IgE and Ca-ionophore A23187. Its activity was lost by heparinase, not by chondroitin-ABC lyase or chondroitin-AC lyase, and also inhibited by Polybrene, suggesting it would be heparin.

Cyclic AMP accumulation in response to norepinephrine was examined in slices of rat cerebral cortex the day after a unilateral application of anodal current of 0.3, 3.0 or 30 microA to the surface of the sensorimotor cortex. When 3.0 microA was applied, the norepinephrine-elicited accumulation of cyclic AMP was greater in the cortical area including the application point than in either the contralateral cortical area or non-polarized control cortical slices. The changes in cyclic AMP accumulation are discussed in relation to the role of the direct current in producing functional changes in the cortex.

A new acidic ninhydrin method for determining free sialic acids is described. The method is based on the reaction of sialic acids with Gaitonde's acid ninhydrin reagent 2 which yields a stable color with an absorption maximum at 470 nm. The standard curve is linear in the range of 5 to 500 nmol of N-acetylneuraminic acid per 0.9 ml of reaction mixture. The reaction was specific only for sialic acids among the various sugars and sugar derivatives examined. Some interference of this method by cysteine, cystine and tryptophan was noted, although their absorption maxima differed from that of sialic acids. The interference by these amino acids was eliminated with the use of a small column of cation-exchange resin. The acidic ninhydrin method provides a simple and rapid method for the determination of free sialic acids in biological materials.

The thalamic posterior ventral neurons with bifurcating axons to both the first and second somatosensory cortical areas (SI and SII) in the cat were examined by the fluorescent retrograde double labeling technique. After injection of Evans blue (EB) into the SI, and of 4',6-diamidino-2-phenylindol.2HCl (DAPI) into the SII of the same hemisphere, EB- and DAPI-labeled cells were observed predominantly in both the posterolateral ventral and the posteromedial ventral nuclei of the thalamus. Although EB single-labeled and DAPI single-labeled cells tended to occupy separate regions within the posterior ventral nuclei, a small number of cells double-labeled with both EB and DAPI were detected in the border zone between two single-labeled cell groups. These observations indicate that some cells in the posteromedial and posterolateral ventral nuclei project both to the SI and SII by bifurcating axons.

Pregnant normal (N) and acatalasemic (A) mice treated with aminotriazole (AT) were exposed to metallic mercury. The mercury contents of the fetus and maternal organs were subsequently determined. The fetal and placental mercury contents were the highest in the AT-treated A mice (A-AT), and the contents decreased in the order of AT-treated N mice (N-AT), non-treated N mice (N-C) and non-treated A mice (A-C). Statistically significant differences in the fetal mercury levels were observed between N-C and A-C, A-C and N-AT, and N-AT and A-AT. The ratios of the mercury concentration in the fetus to that in the maternal blood decreased in the order of A-AT, N-AT, A-C and N-C. The differences in the ratio were significant between these groups. Similar results were obtained when the ratios of the maternal liver level to the maternal blood level or the ratios of the placental level to the maternal blood level were compared. The effect of AT on mercury uptake is remarkable in the fetus of both normal and acatalasemic mice exposed to metallic mercury.

The enzyme activities involved in the transamination of L-cysteine sulfinate (L-alanine 3-sulfinic acid), L-aspartate and L-cysteine were examined in fetal, neonatal and maternal rat liver and placenta. In fetal and neonatal rat liver, aminotransferase activity was most active with L-cysteine sulfinate as a substrate and was also active with L-aspartate, while activity with L-cysteine was very low. The activity of transamination of L-cysteine sulfinate in rat liver developed in parallel with that of L-aspartate and L-cysteine. The aminotransferase activity markedly increased after the 19th day of gestation, reaching the same value as adult liver on the 3rd day after birth. The ratios of transamination of L-cysteine sulfinate to that of L-aspartate and to that of L-cysteine were constant during development. These observations suggest that L-cysteine sulfinate, L-aspartate and L-cysteine are transaminated by the same enzyme in the rat liver during development. Since placental aminotransferase activity was extremely low compared with that of the liver, it was suggested that the placenta did not play an important role in the transamination of these amino acids during pregnancy.

The anti-ulcer action of clotiazepam (a thienodiazepine derivative) was studied in mice subjected to non-physical and physical stimuli in a communication box. There were two groups of mice: the "sender" mice that received electric shocks on the feet and responded by squealing and jumping, and the "responder" mice that were affected by the senders' responses without receiving shocks on the feet. Gastric ulcers resulted in both groups. The effect of clotiazepam was compared with that of diazepam. The incidence of gastric ulcers was suppressed by clotiazepam at a dose of 3 mg/kg, per os, in "responder" and "sender" mice, and by diazepam at a dose of 1 mg/kg, per os, in "responder" mice. These results suggest that clotiazepam has a suppressive action against gastric ulcers produced by non-physical or physical stimuli, although its potency is slightly weaker than that of diazepam.

Concanavalin A (Con A) induced cap formation in rat ascites hepatoma cells (AH7974). In these Con A-treated cells, the association of cytoplasmic proteins with cell membranes was suggested by observing their Triton shells. The transition from G-actin to F-actin occurred in these cells. The association of membrane lipid with cytoplasmic proteins extracted from AH cells was studied by the isolation of protein-bound liposomes and phase transition release. The analysis of isolated liposomes revealed that many cytoplasmic proteins which specifically associated with liposomes were cytoskeletal elements including F-actins. The association of proteins with liposomes was affected by the lipid composition of the liposomal membrane and by the Ca2+ concentration of the incubation medium. The strong interaction of liposomal membrane with cytoplasmic proteins or isolated cytoskeletal proteins was demonstrated also by phase transition release using carboxy fluorescein-containing liposomes. These experiments showed that there was a strong affinity between lipid membrane and cytoskeletal elements including F-actins and that the amount of F-actin increased due to Con A treatment. The association of the submembranous microfilaments with the cell membrane may contribute to capping of the cells caused by Con A.

Titers of antibody against Escherichia coli in human milk and in the sera of 11 breast-fed infants, 6 bottle-fed infants and 9 infants in the post-weaning period were measured by the passive hemagglutination method. High antibody titers were observed in human milk in the first 4 days after parturition, but the titer decreased rapidly thereafter. None of the healthy, breast-fed infants had detectable serum antibodies, while a breast-fed infant with a perianal E. coli abscess had antibodies. On the other hand, 4 of the 6 bottle-fed infants and all of the 9 infants in the post-weaning period had antibodies. The significance of these results was discussed.

The cytostatic and cytotoxic effects of highly purified natural human tumor necrosis factor (HuTNF-alpha) and natural human interferon-alpha (HuIFN-alpha) on 23 cell lines were studied in vitro. Natural HuTNF-alpha showed cytostatic and cytotoxic effects on PC-9, KHG-2, HT-1197, KG-1 and L-929 cells, and HuIFN-alpha showed both effects on KHG-2 and Daudi cells. A mixture of HuTNF-alpha and HuIFN-alpha (1:1, by unit) showed cytostatic and cytotoxic effects on HuTNF-alpha- or HuIFN-alpha-resistant cell lines such as KB, KATO-III, HEp-2, P-4788, as well as on HuTNF-alpha- or HuIFN-alpha-susceptible cells. Thus, the combined preparation of HuTNF-alpha and HuIFN-alpha expanded the spectrum of sensitive cells. The dosage of the mixed preparation required to produce 50% inhibition of cell growth was less than 20% of that of HuTNF-alpha or HuIFN-alpha alone. These results indicate that the cytostatic and cytotoxic effects of HuTNF-alpha and HuIFN-alpha are synergistically enhanced when they are administered together.

Blood vascular beds of fetal, adult and aged human kidneys were reproduced with methyl methacrylate and observed with a scanning electron microscope. The kidney glomeruli, including those from the fetal kidneys, had anastomosing capillaries. The glomeruli in the kidneys of an aged person contained many more capillaries which were much more tortuous than those of the adult and fetal kidneys. Furthermore, it was observed that the glomeruli in the kidneys of the aged person usually received tortuous afferent vessels and frequently emitted multiple efferent arterioles. The glomeruli in the juxtamedullary layer of the kidneys of the aged person were rather small in size and contained degenerative capillary networks. This observation suggests that the medulla of the kidneys of the aged is poorly supplied with blood.

Rat kidney glomerular basement membrane (GBM) was isolated and digested with alpha-amylase and elastase. Electron microscopy revealed a meshwork structure composed of fibrils 3 nm in width. They appeared to be type IV collagen fibrils. We succeeded in clarifying a significant ultrastructural aspect of the GBM which had been unclear until now. The findings are consistent with our previously proposed GBM molecular sieve theory.

We herein determined by fluorescence in situ hybridization the chromosomal localization of 2 human genes, BRAL1 and BCAN, both of which belong to the link-module superfamily, i.e. to the same band of chromosome 1q21-23. Further analysis of the genomic organization of BRAL1 and BCAN revealed that the BRAL1 gene was located 20-kb upstream of the BCAN start site. We isolated a polymorphic dinucleotide (CA) repeat sequence from a genomic clone containing the BCAN gene. High heterozygosity (0.79) makes this polymorphism a useful marker in the study of genetic disorders. Knowledge of the structure of the genes and the marker provides essential information for further analysis of the gene locus at chromosome 1q21-23.