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Twenty-four patients with rheumatoid arthritis (RA) and 20 normal controls were examined for the ability of their peripheral blood B cells to produce interleukin 1 (IL-1) with or without lipopolysaccharide (LPS). B cells were purified from peripheral blood by negative selection methods (i.e., removal of adherent cells and sheep red blood cell rosette-forming cells, followed by treatment with monoclonal antibodies (OKT3 and OKM1) and complement). The amount of IL-1 in B cell culture supernatants (SN) was measured by thymocyte and fibroblast proliferation assays and an enzyme-linked immunosorbent assay for IL-1 alpha and beta. As a group, cultured B cells from patients with RA, both spontaneously and when stimulated with LPS, produced higher levels of IL-1 than those from normal controls. IL-1 production by RA B cells with LPS had a weak but positive correlation with disease activity. Moreover, RA B cell culture SN with elevated levels of IL-1 had a synergistic effect on the growth of anti-human IgM (anti-mu) stimulated B cells. In separate experiments, the growth of RA B cells was significantly promoted by IL-1 beta both with and without anti-mu stimulation. These results suggest that B cell-derived IL-1 may be involved in the B cell clonal expansion of RA through its own activity as a B cell stimulatory factor.

Pulmonary function tests were performed on 234 healthy non-smoking young subjects (189 males and 45 females free from respiratory and allergic symptoms). Maximal expiratory flow-volume (MEFV) curves were visually classified into five MEFV types: Type A, convex or straight flow changes; types B, C, and D, concave-convex-concave flow changes; and type E, sudden flow-fall and accompanying decreased flow rates at lower lung volumes. The reproducibility of MEFV patterns were shown by one way analysis of variance (ANOVA) of MEFV data obtained from 4 groups each consisting of 3-4 males and representing different MEFV types. Distribution of MEFV types was different between males and females; the rate of type A was higher in females than in males and those of types B and E were higher in males than in females. When analyzed in terms of three fractional flow rates, Fr-75, Fr-50, and Fr-25, these values could also be classified into 5 types similarly to the visual MEFV type analysis. It is concluded that MEFV type analysis is useful in assessing health conditions.

We demonstrated the ultrastructure of rat glomerular basement membrane (GBM) by ultra-high resolution scanning electron microscopy. GBM prepared by sonication methods and conductive-staining could be observed without metal coating at magnifications as high as 400,000 times. The GBM showed an irregular meshwork structure composed of various strands and pores. The width of the strands ranged from 6 to 15 nm, and the diameter of pores ranged from 6 to 50 nm. The present study confirmed our molecular sieve theory of the basement membrane.

The cytotoxic effects of ferric nitrilotriacetate (Fe-NTA) have been considered to be caused by free radicals produced by the drug. The present study was carried out to determine whether or not cytotoxic effects of Fe-NTA on cell growth and lipoperoxide formation of Chinese hamster cells were reduced by antioxidants. Using a spin trapping technique, we found that hydroxyl radical formation in the cells increased in the presence of Fe-NTA. Antioxidants, with the exception of superoxide dismutase, slightly inhibited production of the hydroxyl radical. Mannitol significantly reduced lipoperoxide formation, but other antioxidants did not. However, the growth inhibitory effects of Fe-NTA were not attenuated by these antioxidants. These results indicated that the cytotoxic effects of Fe-NTA may be mostly due to unknown factors other than oxygen free radicals.

A 56-year-old man was admitted to our department with a chief complaint of lower extremity dysesthesia. He described a dull numbness below the ankle and a dull pain in the nates for the past two years. Although the numbness extended to the thigh, he did not notice any muscular weakness or atrophy. Neurological examination revealed weakness and atrophy in the face, tongue and the proximal portions of all four extremities. Deep tendon reflexes were decreased. A moderate loss of vibratory sensation was noted below the knees. Electromyography showed neurogenic changes. Muscle biopsy revealed both myogenic and neurogenic changes. Sural nerve biopsy revealed a mild reduction of myelinated fibers, particularly the large-diameter fibers. Based on these findings, a diagnosis of bulbospinal muscular atrophy (BSMA) was made. In recent years, there have been some case reports of BSMA with sensory disturbances, or merely with subclinical manifestations of a sensory disturbance. This case is included in the same category as those reports, but it is interesting to note that the sensory disturbance in the lower extremities occurred as the chief complaint of the disease.

We report a case of a non-Hodgkin's lymphoma (NHL) patient treated successfully with combination chemotherapy during pregnancy who delivered a full-term baby. A 29 year-old patient with cervical and inguinal lymphadenopathy in the 27th week of gestation was referred to our hospital. The diagnosis of lymph node biopsy was NHL (diffuse, large cell type with B-cell phenotype). Three courses of CHOP regimen (adriamycin, cyclophosphamide, vincristine and prednisolone) were given before delivery. The patient has been in complete remission for three years and her baby has been in normal development. Our case supports previous reports that chemotherapy in the third trimester may be given safely on NHL patients.

Neural regulation of the motility between the haustra and taenia coli was studied in the isolated rabbit proximal colon. Four types of haustral and taenial preparations were used: the haustral strip without the taenia coli (type 1), the haustral strip including the taenia coli (type 2), the L-shaped (taenia-haustra) preparations for recording the haustral (circular) response to taenial stimulation (type 3) and the L-shaped (haustra-taenia) preparation for recording the taenial (longitudinal) response to haustral stimulation (type 4). Field electrical stimulation induced a contractile response in the haustra and taenia coli. Hexamethonium reduced the contraction in type 2, 3 and 4 preparations. The desensitization to serotonin reduced the response in type 2 and 3 preparations. After atropinization, the response in types 1 and 4 was reversed to relaxation, and the response in types 2 and 3 was reversed to relaxation followed by contraction which was reduced or abolished by indomethacin. The responses remaining after atropinization in all types of preparations were not affected by other blocking agents tested or desensitization to neuropeptides. Tetrodotoxin abolished all relaxation and contractile responses in all types of preparations. These results suggest that the indirect contractile response to field stimulation is induced mainly via cholinergic and serotonergic neurons, and that the relaxation is mainly mediated by nonadrenergic noncholinergic neurons. The late haustral contractions after atropine may be caused by endogenous prostaglandin.

We examined the effect of fetal calf serum (FCS) on meiotic division, subsequent fertilization, and first cleavage to the 2-cell stage of rat oocytes during in vitro maturation. FCS had no effect on the nuclear progression from dictiate to metaphase of the second maturation in vitro and, FCS had no effect on the first cleavage to the 2-cell stage of fertilized oocytes. However, FCS efficiently increased penetration rate of oocytes and shortened the time required for dissolution of the zona pellucida by alpha-chymotrypsin. These results showed that FCS did not affect cytoplasmic maturation necessary for oocytes to develop to the 2-cell stages. We found that FCS only affects the zona pellucida and does not affect the nucleus or cytoplasm of rat oocytes. FCS may prevent hardening of the zona pellucida.

Ki-67 is a commercially available mouse monoclonal antibody (MoAb), which reacts with a nucleolar antigen (the Ki-67 antigen) expressed in proliferating eukaryotic cells. The author examined the precise localization of the Ki-67 antigen in C-6 cells using immunohistochemical and immunoelectron microscopic methods and estimated the proliferative activity of human brain tumors in situ. Positive nucleoplasmic reactions (early G1 phase) and nucleolar staining (late G1 phase) were observed. The cells showed very weak positive reactions in only one or two nucleoli (S phase) and multiple spicule reactions in the nucleoplasm (G2 phase). During the mitotic phase, the Ki-67 antigen was stained on the surfaces of all chromosomes and finely dispersed in the cytoplasm. By immunoelectron microscopic study, positive reactions were observed on the granular and dense fibrillar components. Therefore, the Ki-67 antigen seems to participate in the processing and assembly of preribosomal particles. In human brain tumors, the Ki-67 score (positive cells/total neoplastic cells), ranging 0 to 36.7%, correlated well with the histopathological grade of malignancy of the tumor. These findings suggest that immunohistochemical staining with the MoAb Ki-67 can be used as a convenient procedure for the simple evaluation of the proliferative activity of brain tumors.

Carbohydrate metabolism of rats with obstructive jaundice caused by bile duct ligation was studied by intravenous glucose tolerance test (IVGTT) and by liver perfusion. The altered levels of carbohydrate-metabolizing enzyme were examined in relation to the glucose metabolism of the cholestatic rats. In the IVGTT, the rate of fractional glucose removal was increased with increases in plasma insulin and glucagon and with a decrease in non-esterified fatty acid. In liver perfusion, neither the glucose uptake nor insulin extraction by the whole liver of icteric rats was different from the control. The increased rate of glucose removal in IVGTT may be due to enhanced glucose utilization by peripheral tissues resulting from hypersecretion of insulin. In liver perfusate supplemented with glucose, a decrease in the glucose uptake per unit liver weight was observed in relation to the lowered glucokinase activity. Formation of glycogen from glucose and of glucose from lactate was also impaired, indicating inhibition of the gluconeogenic system or relative hyperfunction of the glycolytic system, which may further contribute to the reduction in glycogen content. These metabolic disorders correlated well with the changes in activities of key carbohydrate-metabolizing enzymes, which showed a characteristic pattern consistent with the loss of differentiated hepatic functions. Uptake of glucose and its conversion to glycogen were reduced in the cholestatic liver in close association with altered activities of some of related enzymes. However, due to increased utilization by the peripheral tissues, the total amount of glucose utilized in the whole rat was not reduced.

To prevent the development of hepatic metastases after surgery for colorectal cancer, it is important to inhibit the growth of any micrometastases which occur during the operation. We used a hepatic metastasis model in mice to investigate the effects of combination therapy with natural human tumor necrosis factor-alpha (nHuTNF-alpha) and natural murine interferon-alpha/beta (nMuIFN-alpha/beta). Decreased formation of hepatic metastases by murine colon-26 carcinoma was recognized following a single injection of nHuTNF-alpha, nMuIFN-alpha/beta, or both. These inhibitory effects were synergistic. NK activity was also measured, because notaral lerller cells not only have an anti-tumor effect but are also a representative of the host immune system. Both nHuTNF-alpha and nMuIFN-alpha/beta were able to activate NK cells, and the combination of the cytokines more significantly augmented NK activity. The in vivo elevation of NK activity induced by nHuTNF-alpha, nMuIFN-alpha/beta, or their combination may be one of the mechanisms of their antiproliferative effect on experimental hepatic metastases of murine colon-26 carcinoma.

In 15 patients with Japanese cedar pollinosis and 10 healthy control subjects, levels of major basic protein (MBP), eosinophil cationic protein (ECP), and arylsulfatase B (As) in the nasal secretions were examined before and after challenge with Japanese cedar pollen extract. The MBP and ECP levels in the patients were significantly higher 30 min after challenge than those before challenge (P < 0.005). MBP and ECP levels after challenge were significantly higher in the nasal secretions of patients than in the controls (MBP: P < 0.01, ECP: P < 0.05). The level of As after challenge was significantly higher in the nasal secretions of patients than in the controls. These results suggest that eosinophils activate or modify the immediate, nasal allergic reaction and have a role in regulating immunological responses.

To elucidate the effect of the arginine vasopressin (AVP) system in vivo, especially V1 and V2 activity, on blood pressure, we measured the acute changes in blood pressure and heart rate after AVP, OPC-21,268 (a V1 receptor antagonist), and OPC-31,260 (a V2 receptor antagonist) were injected intravenously in anesthetized spontaneously hypertensive rats (SHR) and Wistar-Kyoto (WKY) rats at the age of 15 weeks. Compared with the control period, single injection of AVP 5 ng/kg significantly increased systolic blood pressure in WKY rats without a concomitant increase in heart rate, but there was no significant increase in blood pressure in SHR. In contrast, single injection of either OPC-21,268 3 mg/kg or OPC-31,260 3 mg/kg did not affect blood pressure or heart rate in either SHR or WKY rats. Injection of AVP after the administration of OPC-31,260 induced a greater increase in blood pressure in SHR than in WKY rats, whereas injection of AVP after the administration of OPC-21,268 did not induce any clear increase in blood pressure in SHR or WKY rats. These results suggest that SHR have enhanced pressor activity mediated by V1 receptors and that this increase may be due to an increase in their number. In conclusion, enhancement of V1 activity may contribute to the development of high blood pressure in SHR.

In the fields of psychiatry and neurology, the dopaminergic system is one of the most important neurotransmitter systems in the brain. Whereas pharmacological and biochemical studies had initially indicated two subclasses of dopamine receptors (DA-R), recent progress in molecular biology techniques has led to the identification of five distinct genes of DA-Rs (D1-R-D5-R) and splice variants. The gene products are classified into the D1-R family (D1-R and D5-R) and D2-R family (D2-R, D3-R and D4-R) based on their structure and pharmacological features. This review summarizes the structure, localization, function and pharmacology of DA-R subtypes on the basis of knowledge obtained during the past few years. The genes encoding the D1-R family have no intron and the D2-R family genes have introns. The distributions of mRNAs encoding these five DA-R subtypes in the brain were different from their respective receptors. The localization of DA-R subtypes to particular brain regions and specific pharmacological profiles of DA-R subtypes allow new insights to be made into the mechanism of action of DA in the control of psychiatric and motor functions. The availability of detailed information about DA-R subtypes will not only clarify their roles in the brain, but will probably also lead to the development of new therapeutic drugs with more specific actions.

The establishment of a model system of neoplastic transformation of normal human cells has been attempted with a chemical carcinogen, 4-nitroquinoline 1-oxide (4NQO). In the course of these experiments, it was noticed that immortalization of human cells is a multi-step process involving several mutational genetic events. Thus, chromosomal changes which occurred during the process of immortalization of human fibroblasts were examined. To accomplish immortalization, fibroblasts obtained from an embryo were repeatedly treated with 10_-6M4NQO from primary culture to passage 51 (59 treatments in total). Before immortalization, some chromosomes (especially, chromosomes 2, 6, 8, 10, 11, 12, 15, 19, and 20), were lost at a relatively high frequency. After immortalization, the chromosomes distributed so broadly in the triploid to hypotetraploid region without a distinct modal number or without marker chromosomes that it was difficult to identify the specific chromosomes related to the immortalization of human cells. No specific structural chromosomal changes were detected. Although the significance of such chromosome changes in relation to immortalization is not clear, the loss of some specific chromosomes suggests that genes which are involved in cellular aging and which suppress immortalization may have been lost in the immortalization process.

Tissue PIVKA-II was examined in 32 hepatocellular carcinomas and 2 metastatic liver tumors using indirect immunofluorescence, and the results were compared with the size, histological grading and serum PIVKA-II level. The specificity of this method was confirmed by the disappearance of reactivity in PLC/PRF/5 cells after the addition of vitamin K to the culture medium. Positive PIVKA-II staining was observed as a clustered or a single cell pattern only in the HCC nodules, but not in the surrounding cirrhotic tissue. PIVKA-II staining was observed in all HCC groups regardless of histological grade. There was no relationship between PIVKA-II staining and the size of HCC. PIVKA-II was detected immunohistochemically even in small HCC of patients whose plasma PIVKA-II levels were below the detection limit. These results suggest that PIVKA-II production is a specific phenotype of HCC regardless of its histological grading and demonstrate that this immunofluorescent PIVKA-II staining is more sensitive and useful than plasma PIVKA-II assay for the diagnosis of HCC.

In single treatment study, ethanol was administered intraperitoneally to ICR mice (about 34 g) in the amounts of 1.0, 2.0, 3.0 or 4.0 g/kg body weight. The 3,4-dihydroxyphenylacetic acid (DOPAC) + homovanillic acid (HVA) concentration in the striatum was elevated with 3.0 and 4.0 g/kg of ethanol. In the hypothalamus, the DOPAC, HVA and 5-hydroxyindoleacetic acid concentrations were increased after injection of 3.0 and 4.0 g/kg of ethanol. Furthermore, the acetylcholine (ACh) and gamma-aminobutyric acid (GABA) concentrations were also increased following the injection of 1.0, 2.0, 3.0 and 4.0 g/kg. To study the effects of repeated administration, mice were injected intraperitoneally with 1.0 or 2.0 g/kg of ethanol once daily for 7 days. The DOPAC + HVA level in the striatum was elevated after injection of 1.0 and 2.0 g/kg of ethanol. The GABA and ACh concentrations in the hypothalamus were decreased after repeated injections of ethanol. These results suggest that ethanol significantly alters the utilization of dopamine, ACh and GABA in the hypothalamus. This may partially explain why ethanol has such profound effects on emotional behavior and mood.

Stress distribution in the first carpometacarpal joint was analyzed in 49 cadaveric hands using the finite element method to clarify the pathogenesis of osteoarthritis in the joint. The results of the finite element method analysis were compared with those of the contact pressure distribution in the first carpometacarpal joint of cadaveric specimens using pressure-sensitive film, and with the simple roentgenographical and microradiographical manifestations of spur formation, and with histological findings of osteoarthritis to verify the accuracy of the models of computer simulation models. The comparison of these results showed that osteoarthritic changes of the first carpometacarpal joint were found in areas where stress was concentrated during movement of the joint. The saddle shape of this joint is essentially well-designed for the dispersion of normal stress, however minimal displacement due to instability could easily induce osteoarthritis. Furthermore the shallow trapezial configuration may contribute to the high incidence of osteoarthritis changes. The finite element method helped clarify the relationship between stress patterns and osteoarthritis response.