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The present study was designed to investigate the in vitro effects of potential therapeutic agents on cytokine production by five HTVL-I-infected T cell clones (TCC) established from the ocular fluid of patients with HTLV-I uveitis. Each of the five HTLV-I-infected TCC was cultured at 1 x 10(6) cells/ml with or without an immunosuppressive agent (hydrocortisone, FK506, rapamycin, indomethacin, or prostaglandin E2) for 22 hr in humidified 5% CO2 in air at 37 C. The production of various cytokines in the culture supernatant from each TCC was measured by ELISA. The HTLV-I-infected TCC produced high amounts of IL-1 alpha, IL-3, IL-6, IL-8, TNF-alpha, IFN-gamma, and GM-CSF, and low but significant levels of IL-2 and IL-10 without any stimuli. Hydrocortisone severely depressed the production by these TCC of all the cytokines except for IL-2, which was slightly increased. Prostaglandin E2 depressed the production of IL-1 alpha, while it up-regulated the production of IL-6, TNF-alpha, and IFN-gamma. Rapamycin depressed the production of IL-6 and TNF-alpha, and FK506 depressed the production of TNF-alpha. Hydrocortisone also severely depressed the cytokine production by PHA-stimulated peripheral blood mononuclear cells obtained from healthy volunteers. Of the immunosuppressive agents tested, hydrocortisone exhibited the strongest suppression of cytokine production by HTLV-I-infected TCC. This result was in agreement with the in vivo effects of hydrocortisone in patients with HTLV-I uveitis. These TCC will be useful in investigating the effects of potential therapeutic agents for HTLV-I uveitis in vitro.

Human T-cell lymphotropic virus type I (HTLV-I) is known to cause adult T-cell leukemia/T-cell lymphoma and tropical spastic paraparesis/HTLV-I-associated myelopathy. Recent seroepidemiologic, clinical, and virologic studies indicate that the virus is also related to a certain type of uveitis, which has been classified as uveitis without defined etiologies or idiopathic uveitis. According to the seroepidemiologic survey, the seroprevalence of HTLV-I in patients with idiopathic uveitis was significantly higher than that of two control groups, that is, patients with uveitis with defined etiologies and patients with nonuveitic ocular diseases. Clinically, the uveitis seen in HTLV-I carriers is characterized by moderate to severe cellular infiltration in the eye and by moderate retinal vasculitis, and the intraocular inflammation responds well to corticosteroid therapy. Interestingly, 25% of female patients with the disease had a previous history of Graves disease with hyperthyroidisms. The following virologic, molecular biologic findings suggest that cytokines produced by HTLV-I-infected T cells in the eye play the central role in the pathogenic mechanisms of the uveitis: (a) the virus load in the peripheral blood monocytes analyzed by the quantitative polymerase chain reaction methods was significantly greater in patients with the uveitis than in asymptomatic carriers, (b) the proviral DNA of HTLV-I and the gene expression of the virus at the mRNA level was detected in the infiltrating cells from the eyes of the patients, (c) the virus particles were detected by electron-microscopic examination in the T-cell clones established from the intraocular fluid of the patients, and (d) the HTLV-I-infected T cells produced a variety of cytokines without any stimuli, such as interleukin (IL)-1 alpha, IL-2, IL-3, IL-6, IL-8, IL-10, tumor necrosis factor alpha, interferon-gamma, and granulocyte-macrophage colony-stimulating factor. Based on the seroepidemiologic, clinical, and virologic data, the uveitis seen in HTLV-I carriers is considered to be a distinct clinical entity related to HTLV-I infection, and the disease is designated as HTLV-I uveitis.

Two murine monoclonal antibodies (mAb) designated as NU-T1 and NU-T2 were generated to identify the novel antigens expressed on human T cells. NU-T1 reacted with peripheral blood T cells, but not with granulocytes or monocytes, whereas NU-T2 were not reactive with peripheral blood lymphocytes, granulocytes or monocytes. Both NU-T1 and NU-T2 reacted with thymocytes. The incidence of NU-T1 or NU-T2 reactivity with the leukemic cells from patients with various types of leukemia was as follows: 12/12 or 4/12 in acute lymphocytic leukemia (ALL) of T cell type, 6/6 or 0/7 in adult T cell leukemia, 0/6 or 0/19 in ALL of non-T non-B cell type, and 0/8 or 0/8 in acute myelocytic leukemia, respectively. The incidence of NU-T1 and NU-T2 reactivity with human leukemic T cell lines were 4/10 and 5/10, respectively. In contrast, neither NU-T1 nor NU-T2 reacted with B cell lines, non-T non-B cell lines, or myeloid cell lines tested. These results indicate that NU-T1 and NU-T2 recognize two distinct differentiation antigens expressed on the cells in the T cell lineage. These mAb would be useful reagents to study the function of human T cells and the immunophenotyping of human T cell malignancies.

In serum, the enzyme adenosine deaminase (ADA) is known to be divided into two isoenzymes, ADA1 and ADA2, which have different molecular weights and kinetic properties. The present study investigated ADA isoenzyme levels in the sera of patients infected with retroviruses associated with adult T-cell leukemia (ATL), human T-cell lymphotropic virus type 1 (HTLV-1)-associated myelopathy (HAM), and AIDS, ADA isoenzyme activities were found to be significantly (P < 0.001) higher in the sera of patients with ATL, HAM, and AIDS than in the sera of healthy controls. In the case of the ADA subtypes in the sera of patients with ATL, ADA1 activity was significantly (P < 0.001) elevated in patients with the acute and lymphoma types of ATL compared with that in patients with the chronic and smoldering types of ATL. ADA2 activity was significantly elevated in the sera of patients with the acute, lymphoma, and chronic types of ATL (P < 0.001) compared with that in patients with smoldering ATL and HTLV-1 carriers. In the case of patients with human immunodeficiency virus type 1 (HIV-1) infection, ADA1 and ADA2 activities in the sera of patients with AIDS and HIV-1 antibody-positive individuals were significantly (P < 0.001) higher than those in the sera of HIV-1 antibody-negative individuals. A significant elevation in ADA2 activity was also seen in the sera of AIDS patients (P < 0.01) compared with that in the sera of HIV-1 antibody-positive individuals. These results suggest that the magnitude of elevation of ADA isoenzyme levels in serum correlates well with the clinical conditions of the patients with these diseases. Measurement of the activities of ADA isoenzymes may therefore provide an additional parameter for distinguishing the subtypes of ATL and may prove to be useful as prognostic and therapeutic monitors in diseases associated with HTLV-1 and HIV-1 infections.

This study was performed to evaluate the antimetastatic activity of antitumor agents against metastatic colon carcinoma 26 (Co 26Lu), and to investigate their mechanisms of action. Pirarubicin demonstrated the most striking antitumor activity in mice bearing intravenously injected Co 26Lu cells. Etoposide and mitoxantrone also showed marked antitumor activity. Pirarubicin and mitoxantrone also exerted remarkable inhibitory effect on spontaneous lung metastases from subcutaneously implanted Co 26Lu. Pirarubicin showed marked inhibition of both primary tumor growth and lung metastases. Mitoxantrone was effective in preventing lung metastases even at doses that did not exhibit an antitumor effect on the primary tumor. Moreover, mitoxantrone administered two days after intravenous injection of tumor cells obviously reduced the number of lung colonies, while simultaneous injection of the drug did not inhibit colony formation. Mitoxantrone effectively inhibited angiogenesis on the chorioallantoic membrane at doses that did not affect the growth rate of embryos. These results suggest that mitoxantrone, besides its direct antitumor effect on tumor cells, may inhibit lung metastases by inhibiting angiogenesis.

The MAGE gene family, encoding tumor-rejection antigens recognized by cytotoxic T lymphocytes, is frequently expressed in human solid cancers. However, its expression in leukemia has not been well studied. We have investigated MAGE gene expression at the mRNA level in human leukemia. The MAGE gene family was expressed in 17 of 34 (50%) examples of T cell leukemia (12/21 patients' peripheral blood mononuclear cells and 5/13 cell lines), in 7 of 16 (44%) cases of B cell leukemia (1/8 and 6/8 respectively), but in none of 23 myelomonocytic leukemia cases (0/16 and 0/7), as evaluated by the primers common to the MAGE-1, -3, -4 (-4a and/or -4b), and -6 genes and the semi-quantificative reverse transcription/polymerase chain reaction method. None of a panel of normal lymphoid cells expressed the MAGE gene family. As revealed by the primers specific for each of the MAGE genes, the MAGE-1, -2, -3, -4 or -6 gene was expressed in 8, 8, 6, 2, or 6 respectively out of 23 types of leukemia cell lines. Expression of the MAGE-1 protein in both the cell lines and patients' cells was confirmed by immunoblot analysis with the polyclonal antibody to recombinant MAGE-1 protein. Cellular MAGE-4 protein in the cell lines was measured by an enzyme-linked immunosorbent assay with the polyclonal and monoclonal antibodies to recombinant MAGE-4b protein. In summary, the MAGE gene family was found to be expressed in the substantial proportion of T cell leukemias, but in no case of myelomonocytic leukemia. Antigens coded by the MAGE gene family could be important molecules for understanding specific immunity against lymphocytic leukemia.

A 51-year old woman who had type-C chronic hepatitis developed autoimmune hemolytic anemia after 4 weeks treatment with a total 48 million interferon-beta, 6 million 8 times. Indirect Coomb's test turned from negative to positive through IFN therapy and a complication of asymptomatic primary biliary cirrhosis was confirmed by histologic findings of the biopsied liver which was performed during IFN therapy and a presence of anti-mitochondrial antibody. After stopping IFN therapy and adding prednisolone anemia improved in getting negative indirect Coombs test but remaining positive direct Coombs test.

A 78-year-old male was admitted to our hospital with cutaneous hemorrhage, aphasia and somnolence. On physical examination, he appeared drowsy and complained of epistaxis and multiple purpura. His platelet count was 8,000/microliters with a high level of PA-IgG, but the coagulation time was normal. A CT scan of the head revealed a left temporal and right frontal hemorrhage. Bone marrow aspiration revealed abundant megakaryocytes. He was diagnosed as having idiopathic thrombocytopenic purpura with intracranial hemorrhage. He was treated with prednisolone and azathioprine in addition to platelet infusions and high doses of gamma-globulin. After two months of treatment, the platelet count had risen to 40,000/microliters and his symptoms had subsided.

A human hepatoma cell line constitutively expressing proteins of hepatitis C virus (HCV) was established by transfection with cDNA encoding part of the virus nonstructural (NS) genome region. Proteins consistent with authentic processing at NS3/NS4A, NS4A/NS4B and NS4B/NS5A were identified. Pulse-chase experiments indicated that the cleavage between NS3 and NS4A occurred first and cleavage at other sites followed. Expression of specific surface antigens in response to the presence of HCV proteins was analysed by flow cytometry. A significant increase in CD26 expression was observed in cells expressing the HCV proteins. CD26 plays an important role in cellular signal transduction. Its upregulation in response to the presence of HCV proteins may play a role in viral pathology.

The effects of PGE1.CD on dimethylnitrosamine (DMN)-induced acute liver damage with intravascular coagulation in rats were biochemically and histopathologically investigated. PGE1.CD was administered i.v. from 30 min before to 24 hr after DMN-intoxication (pretreatment) and from 30 min after or from 4 hr after to 24 hr after DMN-intoxication (post-treatment). Pretreatment with PGE1.CD (0.2-2 micrograms/kg/min) dose-dependently suppressed the decrease of platelet counts and the elevation of blood biochemical parameters (PT, HPT, GOT, GPT, LDH, LAP, T-Bil) caused by DMN-intoxication. PGE1.CD (0.5 microgram/kg/min and over) significantly suppressed the DMN-induced histopathological changes (occurrence of hemorrhage and necrosis). Post-treatment with PGE1.CD (2 micrograms/kg/min) also suppressed the liver damage. Furthermore, pretreatment with PGE1.CD (2 micrograms/kg/min) not only suppressed the disruption of hepatocytes, but also prevented the damages of sinusoidal endothelial cells and lysosomal membrane, and it reduced the increase of lipid peroxidation. PGE1.CD (1 microgram/kg/min and over) significantly suppressed the decrease of hepatic tissue blood flow caused by DMN-intoxication. These results demonstrate that PGE1.CD has therapeutically efficacy against DMN-induced acute liver damage in rats; Therefore, it will be clinically useful for the treatment of severe hepatitis such as fulminant hepatitis with intravascular coagulation in the sinusoid.

KD is an acute febrile illness in children characterized by coronary arteritis accompanied by aneurysm and thrombotic occlusion. The etiology of KD is unknown. It has been recently reported that KD is associated with the selective expansion of V beta 2+ and V beta 8.1+ T cells in peripheral blood lymphocytes (PBL), by studying the T cell receptor (TCR) repertoire of in vitro activated T cells. KD may therefore be caused by a superantigen [1-3]. To understand better the immunopathology of KD, we investigated TCR V beta 2 and V beta 8.1 expression on both the T cells of freshly isolated PBL and T cell clones (TCC) from patients with KD. Cytokine production by TCC was also studied. Blood samples were obtained from patients with acute (n = 20) and convalescent (n = 20) KD, age-matched children with non-infectious diseases (n = 18), and healthy adults (n = 20). Among these four groups, there were no significant differences in the percentages of either V beta 2+ or V beta 8.1+ T cells of freshly isolated PBL. The same was true for the CD4+ or CD8+ T cell subsets. One hundred and five TCC (98 CD3+ CD4+ CD8- and seven CD3+ CD4- CD8+) established from the affected skin, lymph node or PBL of six patients with KD were also negative for either V beta 2 or V beta 8.1 TCR. Sixty-eight of 105 TCC (65%) produced detectable levels (> 5 pg/ml) of TNF-alpha (6-1016 pg/ml), in the absence of any stimuli. In contrast, only 11 (10%) of 105 TCC or 7 (7%) of 97 TCC produced detectable levels of IL-2 or IL-6, respectively, in the absence of any stimuli. Stimulation with phytohaemagglutinin (PHA) and phorbol myristate acetate (PMA) induced most TCC to produce higher amounts of TNF-alpha, IL-2 and IL-6. These results suggest that CD4+ T helper cells expressing TCR-beta other than V beta 2 or V beta 8 receptor, primarily through TNF-alpha production, are involved in the immunopathology of KD.

The immunopathology of human T cell lymphotropic virus type 1 (HTLV-I) uveitis was addressed by using T cell clones (TCC) established from the intraocular fluid of patients with HTLV-I uveitis. Proviral DNA of HTLV-I was identified in 55 out of 94 (59%) or 13 out of 36 (36%) TCC from the ocular fluid or the peripheral blood of these patients, respectively. Most of HTLV-I-infected TCC had a CD3+ CD4+ CD8- phenotype. HTLV-I infection on TCC was confirmed by analysis of the viral mRNA, nucleotide sequence, virus-associated proteins, and virus particles. HTLV-I-infected TCC, but not HTLV-I negative TCC, constitutively produced high amounts of IL-6 (1,336 +/- 1,050 pg/ml) and TNF-alpha (289 +/- 237 pg/ml) in the absence of any stimuli. HTLV-I-infected TCC from the ocular lesion also constitutively produced high amounts of IL-1 alpha (12,699 pg/ml), IL-2 (61 pg/ml), IL-3 (428 pg/ml), IL-8 (1,268 pg/ml), IL-10 (28 pg/ml), IFN-gamma (5,095 pg/ml), and GM-CSF (2,886 pg/ml). Hydrocortisone, a drug effective in vivo for the treatment of HTLV-I uveitis, severely depressed cytokine production in vitro in most cases. In summary, the results demonstrated direct evidence of HTLV-I infection of the eye and suggest that cytokines produced by HTLV-I-infected T cells are responsible for the intraocular inflammation in patients with HTLV-I uveitis.

Purpose: To understand better the immunopathology of HTLV-I uveitis by investigating the clonality of HTLV-I-infected T-cell clones., Methods: Eleven T-cell clones were established from the aqueous humor (six clones) and the peripheral blood (five clones) of a patient with HTLV-I uveitis, and the clonality of the HTLV-I-infected T cells was investigated by sequencing the T-cell receptor (TCR) alpha gene after the amplification of TCR alpha cDNA using an adaptor-ligation method and reverse transcriptase-polymerase chain reaction (RT-PCR)., Results: TCR alpha use was different for each of 11 T-cell clones, encompassing eight different HTLV-I-infected T-cell clones (four from the aqueous humor and four from peripheral blood) and three HTLV-I-negative T-cell clones., Conclusions: This study demonstrated polyclonal use of TCR alpha for HTLV-I-infected T cells in the ocular lesion and the peripheral blood. Results suggested that these T cells are not precursors of the leukemic cells associated with malignant transformation. Instead, they might be randomly infected with HTLV-I in the process of HTLV-I uveitis.

Behçet's disease is a systemic inflammatory disease immunologically characterized by the infiltration of CD(4)(+)T cells and polymorphonuclear cells in the affected tissues, including the eye. Despite extensive studies, neither the etiology nor the pathogenic mechanism of Behçet's disease has been clarified. Production of cytokines by T cell clones (TCC) from the ocular fluid (18 CD(3)(+)CD(4)(+)CD8 clones and 4 CD(3)(+)CD(4)(-)CD8(+) clones) and peripheral blood mononuclear cells (PBMC) (16 CD(3)(+)CD(4)(+)CD8(-) clones) of two patients with uveitis associated with Behçet's disease was investigated to understand better the immunopathology of the disease. The level of IL-8 spontaneously produced by TCC from the ocular fluid (mean: 659 pg/ml) or PBMC (536 pg/ml) of the patients was significantly higher (p<0.005, and p<0.05, respectively) than that by TCC (18 CD(3)(+)CD(4)(+)(-)CD8(+) clones) and 4 CD(3)(+)CD(4)(-)CD8(+) clones) from PBMC of healthy donors (241 pg/ml). As for the other cytokines, IL-Iα, IL-2, IL-6, IL-10, and TNF-α were produced at low but detectable amounts by the majority of TCC from the ocular fluid of patients with Behçet's disease; IL-3, IFN-Y, and GM-CSF were produced by some of TCC at low amounts; and IL-4 was not produced by any TCC tested. IL-8 production by TCC from the ocular fluid was further up-regulated upon stimulatation with PHA, but was suppressed by FK506 and hydrocortisone, though not by diclofenac sodium. Colchicine rather increased IL-8 production by these TCC. These results suggest that T cells in both the affected tissues and PBMC play an important role in the pathogenesis of uveitis associated with Behçet's disease through IL-8 and probably the other cytokines.

Proximal tubular reabsorption is of primary importance in the regulation of the homeostasis of inorganic sulfate, an electrolyte that is necessary for biosynthetic and detoxification reactions. The objective of the present investigation was to determine the effect of dietary sulfate deprivation, produced by a diet low in methionine, on the renal transport of sulfate. Female Lewis rats were fed a diet containing negligible amounts of sulfate and cystine and low in methionine (0.37%) or a control diet (methionine 1.12%, cystine 0.07%) for 8 days to examine the urinary excretion rate and renal clearance of sulfate. The sulfate excretion rate was decreased by day 4 of the low methionine diet and remained low. Both the urinary excretion rates and renal clearance values were significantly decreased on day 8 (144 +/- 71 vs. 517 +/- 264 mumol/12 hr in controls, mean +/- S.D., n = 7, P < .005 and 0.38 +/- 0.19 vs. 1.07 +/- 0.61 ml min-1 kg-1 in controls, n = 5-6, P < .05, respectively), although the serum sulfate concentrations were unchanged. In vitro transport studies were performed in kidney cortex brush border membrane (BBM) and basolateral membrane vesicles isolated from rats fed either the low methionine or control diet for 7 to 9 days. The Vmax for BBM sodium/sulfate cotransport was increased in kidneys from animals that received the low methionine diet (1.1 +/- 0.10 vs. 0.75 +/- 0.08 nmol mg of protein-1 10 sec-1 in controls, n = 5, P < .001); there were no significant differences in the Km.(ABSTRACT TRUNCATED AT 250 WORDS)

A novel human T cell line (SALT-3) was established from the pleural effusion of a patient with adult T cell leukemia (ATL) of lymphoma type. SALT-3 showed atypical T cell markers such as CD1-CD2-CD3-CD4+CD5+CD7+CD8-CD19-CD20-CD25+HLA-DR+. T cell receptor alpha/beta and gamma/delta were undetectable. Human T cell lymphotropic virus type 1 (HTLV-I) particles were seen on SALT-3 cells by electron microscopic analysis. HTLV-I gag p19, proviral DNA and mRNA of HTLV-I genes were also detected in the cells. Chromosome analysis showed abnormal karyotypes as 47, XY, partial trisomy of No.3 chromosome, and trisomy of No. 7 chromosome. Furthermore, SALT-3 were susceptible to the infection of human immunodeficiency virus type 1 (HIV-1) and the cells were rapidly killed after HIV-1 infection. This newly established HTLV-I-infected human T cell line would be a useful tool to study biological activities of atypical type of ATL cells and to examine the cytotoxic effects of HIV-1 and it's modulators.

To investigate the regulation of endothelin-1 (ET-1) production, cardiac catheterization was performed in young patients with congenital heart disease (27 +/- 5 months old, mean +/- SEM), and plasma levels of ET-1 in the inferior vena cava were measured by a sandwich-enzyme immunoassay. Plasma ET-1 levels of age-matched healthy controls were 1.55 +/- 0.07 pg/ml (n = 6). In patients with atrial septal defect [without pulmonary hypertension (PH)] who had volume but not pressure overload to the pulmonary circulation (PC), plasma ET-1 levels were significantly lower than in controls. In patients with PH due to ventricular septal defect (VSD) who had both volume and pressure overload to PC, the levels were significantly higher than in controls. In patients with PH and severe pulmonary congestion due to pulmonary venous stenosis (PVS), plasma ET-1 levels were significantly higher than in those with VSD, suggesting that ET-1 production is also augmented by pulmonary congestion. The present findings suggest that ET-1 production is increased by pressure overload to PC but decreased by volume overload to PC.

In an attempt to identify the antigens expressed on human myeloid leukemia cells, two murine monoclonal antibodies (mAb) designated as MCS-1 (isotype; IgG3) and MCS-2 (IgG1) were raised. MCS-1 reacted with peripheral blood granulocytes, but not with monocytes, whereas MCS-2 reacted with both granulocytes and monocytes. The incidence of MCS-1 and MCS-2 reactivity with the cells from a total of 121 patients with various type of leukemias was as follows: 25/46 (54%) and 39/47 (83%) in acute myeloblastic leukemia and acute monoblastic leukemia, 8/16 and 16/16 in chronic myeloid leukemia in blastic crisis (CML-BC) of myeloid type, 0/7 and 2/7 in CML-BC of lymphoid type, 0/26 and 2/32 in acute lymphoblastic leukemia (ALL), 7/7 and 7/7 in chronic phase of CML, respectively. Two cases of MCS-2 positive ALL had a Philadelphia chromosome marker (Ph1). In contrast, neither MCS-1 nor MCS-2 reacted with lymphocytes or any of leukemic and non-leukemic lymphoid cell lines tested. These results indicate that MCS-1 and MCS-2 mAb recognized two different differentiation antigens expressed on granulocytes and monocytes. These mAb would be useful reagents to determine differentiation antigens on the cells in granulocyte and monocyte lineage.

Inorganic sulfate is a physiological anion which is utilized in the metabolism of both endogenous compounds and xenobiotics. Its homeostasis is maintained predominantly by facilitated reabsorptive processes in the kidneys. The objectives of the present investigation were to evaluate the effects of menopausal status and caffeine ingestion on the serum concentrations and clearance of inorganic sulfate. Thirty-nine women who were classified as premenopausal, postmenopausal with or without estrogen treatment, and postmenopausal with osteoporosis participated in the study. The women were studied on two separate occasions following the ingestion of a decaffeinated beverage to which 6 mg caffeine/kg lean body mass or no caffeine was added. All women were habitual caffeine users (mean ingestion of 588 mg caffeine per day) but abstained from all caffeine sources for 2 weeks prior to the control study day. Postmenopausal women with estrogen supplementation exhibited significantly lower sulfate serum concentrations (0.24 +/- 0.02 mM vs. 0.32 +/- 0.04 mM in premenopausal women, mean +/- SD, p < 0.05) and a decreased renal reabsorption of sulfate for the control (no caffeine) period. There was no difference in serum sulfate or sulfate reabsorption in estrogen supplemented postmenopausal women, compared with women not taking estrogen. Postmenopausal women with osteoporosis had significantly lower creatinine and sulfate clearances than postmenopausal women with estrogen supplementation which may be related to their older age, or factors related to the disease process. The 6 mg/kg dose of caffeine caused a diuresis, but no change in GFR, as indicated by urine volume and creatinine clearance values, respectively. Caffeine administration resulted in an increase in the sulfate excretion rate; there was no change in sulfate serum concentrations. The results of this investigation indicate that menopause results in decreased sulfate serum concentrations that may be the consequence of a decreased renal reabsorption of sulfate. Secondly, this investigation demonstrated that caffeine ingestion increases the urinary excretion of sulfate, an effect that may be related to the diuretic effect of caffeine or due to a caffeine-induced alteration in the renal reabsorption of sulfate.

Data on T-cell subsets from 89 human T-cell lymphotropic virus-I (HTLV-I) carriers and 25 seronegative people were analyzed to identify differences in T-cell subset values among three subgroups: HTLV-I carriers with abnormal lymphocytes (Ably; n = 24), carriers without Ably (n = 65), and HTLV-I seronegatives (n = 25). Estimates of mean values were adjusted for age, sex, smoking, and alcohol drinking, as appropriate. The percentage of CD25+ T cells was elevated in carriers with Ably (mean, 16.7 +/- 1.0) compared with the seronegatives (11.4 +/- 1.4; p = 0.0002); individuals with CD25 T-cell percentages above the median for the seronegatives had a corresponding 5.4-fold risk for being a carrier with Ably. Similarly, the percentage of CD4 T cells was elevated in carriers with Ably. Conversely, the percentage of CD8 T cells was lower among both groups of HTLV-I carriers than in the seronegatives. There was a corresponding significant increase (p = 0.0004) of the CD4/CD8 ratio among carriers with Ably (1.57 +/- 0.12) compared with the seronegatives (1.22 +/- 0.12). Among subjects with CD4/CD8 ratios above the median for the seronegatives, there were 6.8- and 4.5-fold risks for being carriers with or without Ably, respectively. The percentage of CD7 was lower among carriers with Ably (75.6 +/- 1.6) than among seronegatives (78.9 +/- 1.5; p = 0.13). The percentage of beta-interleukin-2-receptor-positive T cells did not vary among the three subgroups.(ABSTRACT TRUNCATED AT 250 WORDS)

Objectives and Design: The expression of the accessory molecule CD28 was compared in various populations of T and natural killer (NK) cells from HIV-1-negative and HIV-1-positive individuals and correlated with activation using mitogens in vitro., Methods: Multiparameter flow cytometric analysis using combinations of CD3 CD28 and other markers was performed together with absolute cell counting in peripheral blood. Blast transformation and proliferative responses were also quantitated using the Cytoronabsolute after stimulation with phytohaemagglutinin (PHA) and anti-CD3. CD28- cells were also purified to confirm the observations., Results: In HIV-1-negative individuals > 90% of CD3+ T cells were CD28+ and responded to stimulation, while CD3- CD16+ CD57+ NK-like cells were CD28- and failed to respond. In HIV-1-positive individuals the expression of CD28 was greatly reduced and the proportion of CD3+CD28- T cells expanded. CD8 lymphocytosis was caused entirely by the accumulation of CD28- T cells and many of these expressed activation markers human lymphocyte antigen-DR, CD38 and CD45RO on their membrane and molecules such as TIA-1 and perforin, associated with cytolytic function, in their cytoplasm. The strong positive correlation (r = 0.66) between the lack of CD28 expression and the poor proliferation from HIV-1-positive individuals was confirmed by demonstrating that only CD28+ cells transformed into lymphoblasts and proliferated. Although the CD28- including CD3+ T cells transiently expressed CD25 (interleukin-2R alpha), they did not undergo blastogenesis or activation measured by bromodeoxyuridine uptake and died after 3-4 days in culture. These observations were confirmed in costimulation experiments with anti-CD2 and anti-CD28., Conclusion: In HIV-1 infection activated CD3+CD28- T cells accumulate but are unresponsive to mitogens and anti-CD28. These cells appear to represent terminally differentiated effector cells which fail to respond to further stimuli because of the absence of a CD28 second signal.

A highly specific antiserum against rat liver metallothionein (MT) was raised in a Japanese white rabbit. Using this anti-MT antiserum, we found that MT was localized in the nuclei as well as in the cytoplasm of hepatocytes in newborn rats. Since it is known that these cells are growing actively, we suspected that there was a relationship between the localization of MT in cell nuclei and the cell proliferation. Therefore, the induction and subcellular localization of MT were examined in rat liver remaining after 70% removal. MT was induced in the remnant liver rapidly after the hepatectomy, its concentration being about 80-fold higher than that of the intact liver. Flow cytometric analysis revealed that MT was translocated into the nuclei from the cytoplasm of hepatocytes during liver regeneration after partial hepatectomy. The highest MT level in the nuclei was observed 24 h after hepatectomy. MT-stained positive nuclei were in S to G2M phases of the cell cycle of regenerating hepatocytes, and the nuclei in G1 phase were not stained with anti-MT antiserum. The increase in hepatic MT levels did not directly cause MT translocation into the nuclei. These results suggested that MT was a cell cycle-dependent, nuclear protein.

Two cDNA fragments of terminal deoxynucleotidyl transferase, which contained new insertion sequences, were found from bovine thymus cDNA. One of the clones encoded 18 new amino acids and the other encoded 9 new amino acids. In the results of genomic structure analysis around the new insertion sequences, alternative splicing of the bovine terminal deoxynucleotidyl transferase gene was suggested.

In normal T cells, an anti-CD28 MoAb (Kolt-2) will synergize with the mitogenic stimuli phytohaemagglutinin (PHA), anti-CD3 (OKT3) or a combination of anti-CD2 antibodies (OKT11 and GT2) in the induction of DNA synthesis. A subgroup of patients with common variable immunodeficiency (CVID) show a defect in DNA synthesis by T cells stimulated in vitro with the above mitogens. We have now investigated whether anti-CD28 will correct the defect. This strategy partially restored DNA synthesis, providing evidence that the CD28 co-stimulatory pathway in CVID T cells is normal. Ligation of CD28 acts through co-stimulating IL-2 secretion. The natural ligand (B7) for CD28 on antigen-presenting cells from CVID patients is expressed normally. We conclude that the defect in CVID T cells lies in pathways that lead to transcription of the IL-2 gene other than that induced by ligation of CD28 with Kolt-2.

Surface molecules involved in human T cell activation were investigated using a newly developed monoclonal antibody (H47 mAb). H47 antigen (Ag) recognized by H47 mAb was expressed on approximately 10% of resting T cells (mostly CD4-CD8+), 30% of phorbol 12-myristate 13-acetate (PMA)-activated T cells (both CD4+CD8- and CD4-CD8+), and most NK, B cells, and monocytes in the peripheral blood mononuclear cells (PBMC). H47 mAb respectively immunoprecipitated a 100 or 120-kD molecular weight (MW) membrane protein of T cells and monocytes under nonreducing or reducing conditions, suggesting that H47 Ag consists of a single polypeptide that has intramolecular disulfide bonds. H47 mAb significantly enhanced PMA-induced proliferation of PBMC in a monocyte-independent fashion. H47 mAb, however, failed to enhance T cell proliferation induced by anti-CD3 mAb, anti-CD2 mAb, or phytohemagglutinin (PHA). H47 mAb also enhanced PMA-induced interleukin-2 receptor (IL-2R) expression and IL-2 synthesis, but did not induce a change in intracellular free calcium ([Ca2+]i) of T cells. These results suggest that H47 Ag is a new membrane molecule involved in PMA-induced T cell activation.

We developed and characterized a monoclonal antibody (mAb), H227, recognizing a new epitope of human 4F2 molecular complex whose roles remain to be identified. Both staining patterns and molecular sizes recognized by H227 mAb were the same to those of 4F2 mAb. Thus, both H227 and 4F2 mAbs were reactive to monocytes, thymocytes, and activated lymphocytes and immunoprecipitated the 125-kDa molecular weight membrane protein under nonreducing conditions and 85-kDa heavy-chain and 40-kDa light-chain proteins under reducing conditions. These two mAbs immunoprecipitated only the 85-kDa heavy-chain protein when the cell lysate was initially treated with dithiothreitol. Sequential immunoprecipitation proved their cross-reactivity. Both the mAbs inhibited phytohemagglutinin- or concanavalin A-induced proliferation of peripheral blood mononuclear cells (PBMC). In contrast to these similarities, the pretreatment of cells with one mAb failed to block the reactivity to the other, suggesting that their epitopes were different from each other. Furthermore, only H227 mAb augmented phorbol myristate acetate (PMA)-induced PBMC proliferation in association with increase in interleukin 2 receptor expression. In summary, H227 mAb recognizes a new epitope of 4F2 heavy chain that might be involved in the PMA-induced T cell activation pathway and therefore shall be a useful tool for understanding the roles of the 4F2 molecular complex.

Nef is one of the non-structural genes encoded by human immunodeficiency viruses. The protein product is associated with the cellular and plasma membranes, and is synthesized soon after entry of the virus. However, little is known about its functions relevant to viral replication and cytopathogenesis. CD4 is considered to be a crucial molecule for the signal transduction and maturation of T cells, and also serves as the receptor for HIV-1. It has been suggested that the down-regulation of cell surface CD4 by Nef proteins is somehow controversial. In order to evaluate the effects of Nef on the CD4 molecule, we constructed a CD4+ monocytoid cell line in which the HIV-1 nef gene was placed under the transcriptional control of the human metallothionein IIA promoter. The analysis of this cellular clone showed that CD4 on the cell surface was down-regulated when Nef proteins were induced. The amount of CD4 antigens on the cell surface correlated inversely with the level of Nef protein expression. This down-regulation of CD4 antigens by Nef was found to occur at the post-translational level. These results showed that the nef gene could modulate the pathophysiology of AIDS by altering the surface CD4 expression.

Distribution of CD11b Ag, a member of the leukocyte integrin (beta 2 integrin) family, on human T cells is not well characterized. We investigated in this study the tissue distribution and age-related alteration of human CD11b+ T cells. Significant proportions of CD11b+ cells were found in T cells from adult peripheral blood (18-58 yr) (20%), liver (32%), and spleen (16%), but were rarely observed in those from thymus, lymph nodes, intraepithelial lymphocytes, lamina propria lymphocytes, and lung. In contrast, percentages of CD28+ T cells, a counter population of CD11b+ T cells, were highest in lymph nodes (91%), modest in intraepithelial lymphocytes (82%), lamina propria lymphocytes (77%), PBL (77%), spleen (73%) and lower in thymus (62%), lung (60%) and liver (56%). The CD11b+ T cells from peripheral blood gradually increased with age, whereas the CD28+ T cells showed an age-dependent decrease. Namely, the percentages of CD11b+ T cells were 6% in newborns (cord blood), 15% in young adults (18 to 22 yr), 25% in middleaged adults (45 to 58 yr), and 27% in aged adults (72 to 86 yr), whereas those of CD28+ T cells were 96%, 83%, 72%, and 71%, respectively. Proportions of CD11b+ T cells in TCR-gamma delta+ or CD8+ T cells were, respectively, higher than those in TCR-alpha beta+ or CD4+ T cells. Age-dependent increase of CD11b+ T cells in CD8+ T cells was more obvious than that of CD4+ T cells. Furthermore, CD11b+ CD28+ T cells increased in the peripheral blood only of aged adults. The percentage of CD11b+ CD28+ T cells in TCR-gamma delta+ T cells was highest in young adults (25%), whereas that in CD4+ T cells was highest in aged adults (15%). CD11b+ CD28+ T cells expressed much potent IFN-gamma, IL-2, and IL-4 at mRNA level than CD28+ CD11b- T cells or CD11b+ CD28- T cells, and had potential cytotoxic activity. Roles of CD11b+ T cells in vivo were discussed.

The effects of combination therapy including various antitumor agents and interferon on mice bearing hepatic metastases of colon carcinoma 26 were determined. Combined treatment with interferon-alpha A/D and carboplatin (CBDCA) was associated with a considerably more pronounced antitumor effect than was treatment with either drug alone. Murine interferon-beta and -gamma each also potentiated the antitumor activity of CBDCA. Combination therapy with interferon-alpha A/D and CBDCA also resulted in marked inhibition of hepatic metastasis of M5076 reticulum cell sarcoma. However, interferon-beta did not potentiate the antitumor activity of CBDCA against either subcutaneously implanted colon carcinoma 26 or pulmonary metastases of this tumor. Thus, in our model the combined administration of interferon and CBDCA was associated with a synergistic antitumor effect on hepatic metastases alone.

To investigate the pathogenesis of Kawasaki disease, we examined biopsy specimens from the skin lesions of 10 Japanese patients at the site of the polymorphous exanthem on the hip and at the site of an inoculation with BCG (bacillus Calmette-Gruérin) vaccine of two of them. Ten normal patients were also examined for control purposes. The histopathologic features of the skin lesions were characterized by extensive edema with dilation of the small blood vessels in the papillary dermis; the site of BCG vaccine inoculation showed much greater inflammatory change; neutrophil emigration was weak and most of the infiltrates were CD4+ T lymphocytes and CD13+ macrophages. The expression of the DR locus of human leukocyte antigen was detected not only on the epidermal keratinocyte surface but also on the walls of the small blood vessels and the infiltrating cells around these blood vessels. We also detected cytokines in the skin lesions: (1) interleukin-1 alpha and tumor necrosis factor alpha were strongly positive in all patients with acute Kawasaki disease, (2) interleukin-2 and interferon gamma were weakly or partially positive, (3) no cytokines were detected in the convalescent phase, and (4) the amounts of cytokines at the site of BCG vaccine inoculations were larger than those at the site of the polymorphous exanthem. These findings suggest that CD4+ T lymphocytes and CD13+ macrophages are activated and interleukin-1 alpha and tumor necrosis factor alpha may be involved in the pathogenesis of the inflammation of acute Kawasaki disease.

Although pentobarbital has been found to depress myocardial function, the magnitude of its direct effects on ventricular contraction at anesthetic concentrations has not been well quantified. The direct effects of pentobarbital on left ventricular function were measured by employing an isolated canine heart preparation with a blood oxygenator. Seven hearts were perfused with blood, dextran, and perfluorochemical artificial blood. Ventricular function was evaluated using the slope of the end-systolic pressure-volume relationship (Ees) and the maximal rate of pressure development (dP/dtmax) in ventricles contracting isovolumically in control, after a low dose (13 micrograms/ml), and after a high dose (48 micrograms/ml) of pentobarbital. These concentrations represent one-half and two times the typical value (25 micrograms/ml) found to produce anesthesia in canines (assessed by tail clamp or blink reflex). The low dose of pentobarbital did not produce clear-cut depression in contractile function. The high dose of pentobarbital produced significant reductions of Ees, and dP/dtmax: Ees decreased 29%, from a control of 4.30 +/- 0.84 to 3.05 +/- 0.49 mmHg/ml and dP/dtmax decreased 24%, from a control of 909 +/- 148 to 695 +/- 173 mmHg/s. Thus, the threshold for the direct depressant effect of pentobarbital on ventricular function falls within the range of half to double the typically-reported anesthetic concentrations.

CD1 antigens are classified into at least three groups, CD1a, CD1b, and CD1c. In order to delineate the localization of epitopes of CD1 antigens in human skin, we examined the immunoreactivity of fourteen different CD1 antibodies (seven CD1a, five CD1b, and two CD1c antibodies). The epitopes for CD1a, CD1b, and CD1c are differentially localized on epidermal Langerhans cells, dermal dendritic cells, keratinocytes, the luminal portion of eccrine gland ducts, and the basement membrane zone in normal human skin.

Background: CD1 antigens are classified serologically into at least three groups, CD1a, CD1b, and CD1c, and many kinds of monoclonal antibodies are available for each subgroup of CD1 antigens. CD1a, CD1b, and CD1c antigens have been shown to be selectively and differentially expressed on epidermal Langerhans cells and dermal dendritic cells in normal human skin., Objective: The objective was to further delineate the localization of epitopes of CD1 antigens in human skin., Methods: We examined the immunoreactivity of 14 different CD1 antibodies (seven CD1a, five CD1b, and two CD1c antibodies) with the immunoperoxidase technique. We also studied the reactivity of NU-T2 (CD1b) antibody by immunogold electron microscopy., Results: The epitopes for CD1a, CD1b, and CD1c antigens were differentially mapped on epidermal Langerhans cells, dermal dendritic cells, keratinocytes, the luminal portion of eccrine gland ducts, and the basement membrane zone in human skin., Conclusion: These CD1 antibodies may be useful to analyze the phenotypic alteration of immune and nonimmune cells in various skin diseases.

Ten patients with chronic lymphoid leukemia were analyzed for clinical characteristics, morphology and phenotype of leukemic cells. There were 3 patients with T-chronic lymphocytic leukemia (CLL), 2 with T-prolymphocytic leukemia (PLL), 2 with B-CLL, 1 with B-PLL, 1 with non-T-non-B-CLL and 1 with Waldenström's macroglobulinemia. Although chronic lymphoid leukemia is usually characterized by proliferation of B-lymphocytes, our study revealed that 5 of 10 patients had T-cell phenotype. A peripheral blood specimen of T-CLL showed small lymphocytes with a mature appearance and an irregular nuclear margin. Most of the cells lacked large azurophilic granules in the cytoplasm and nucleoli were also inconspicuous in the nucleus by light and electron microscopic examinations. In PLL, a majority of the cells were large lymphocytes with a prominent nucleolus and abundant basophilic cytoplasm which were more clearly observed by transmitted electron microscopic examination. The patients were treated with cyclophosphamide and prednisolone, but response to treatment was transient. The median survival time was 7 months for all patients, while that of T-cell lineage cases was only 1 month. Therefore, new modalities of treatment must be investigated in the future.

Clinically useful monoclonal antibodies, applied for immunophenotyping of leukemias, are reviewed. With a combination of 15 antibodies, including CD2, CD3, CD4, CD5, CD7, and CD8 for T cell marker analysis, CD10, CD19, CD20, surface immunoglobulins, and cytoplasmic mu chain for B cell marker analysis, CD13 and CD33 for myeloid marker analysis, and HLA-DR and CD25 for other marker analysis, acute lymphoblastic leukemias of T cell type, cALL type, pre-B cell type and B cell type, acute myeloid leukemias, acute unclassified leukemias and adult T cell leukemias could be clearly diagnosed by immunophenotyping of cell membrane molecules. By using additional CD11b, CD14, and CD15 monoclonal antibodies, subclassification of acute myeloid leukemia was partially possible.

We investigated the origin of leukemic progenitors in a case of the simultaneous occurrence of myelomonocytic leukemia and multiple myeloma (IgG-kappa). At presentation, myeloperoxidase and nonspecific esterase-positive myelomonocytic cells had proliferated up to 12.2 x 10(9)/liter in the peripheral blood. Bone marrow cell differentials revealed the coexistence of myelomonocytic cells (30%) and atypical plasmacytoid cells (26%). Myelomonocytic cells in peripheral blood expressed both myeloid antigens (CD11b, CD13, CD14, CD15, CD33) and T/B-lymphoid antigens (CD2, CD4, CD5, CD7, CD10, PCA-1). Bone marrow mononuclear cells (BMMC) could be divided into PCA-1 strongly positive and PCA-1 weakly positive populations, which were considered to represent myeloma cells and myelomonocytic cells, respectively; the former were CD2-positive (CD2+), CD14-, and CD15-, whereas the latter were CD2+, CD14+, and CD15+. Immunohistochemical analysis revealed that, in addition to plasmacytoid cells, a minority of myelomonocytic cells showed a positive reaction for IgG staining, and production of IgG was observed in the culture supernatant of CD14+ myelomonocytic cells in peripheral blood. Southern blot analysis revealed the presence of two identical rearrangement bands of immunoglobulin heavy chain gene in both BMMC containing myeloma cells and myelomonocytic cells and CD14+ myelomonocytic cells in peripheral blood. In a long-term methylcellulose assay, peripheral blood mononuclear cells produced large compact colonies consisting of macrophages and IgG+ plasmacytoid cells (M phi/P colonies), while BMMC produced a different type of colonies consisting of CD14+ myelomonoblasts, macrophages, and IgG+ plasma cells (Mb/M phi/P colonies) in addition to M phi/P colonies. Recloning experiments showed that primary Mb/M phi/P colonies gave rise to both secondary M phi/P and Mb/M phi/P colonies. These observations strongly suggest that common leukemic progenitors provide both myeloma and myelomonocytic leukemia cells, and the mechanism of "lineage infidelity" is probably involved in the development of their "bilineal" differentiation.

We examined 24 pediatric patients to evaluate the usefulness of sudden near maximal exercise test (dash method), where the subjects began to run at Bruce protocol of the last stage. 1) No considerable differences between two protocols were found in maximal oxygen consumption (V O2max), maximal heart rate (HRmax), maximal systolic blood pressure, and findings of electrocardiography. 2) The sudden maximal exercise test could be completed during a shorter period compared to Bruce protocol. 3) The sudden maximal exercise protocol reached 84% of HRmax and 47% of V O2max at one minute after the onset of the protocol, and produced 96% of HRmax and 89% of V O2max at two minutes after the onset. We considered that sudden maximal exercise protocol was useful to obtain a response similar to Bruce protocol at maximal exercise within a short period. We have to pay attention to the safety of the patients because their cardiopulmonary response to sudden maximal exercise protocol is dramatic.

To determine the dynamic property of cardiac function response to alterations in coronary perfusion, we varied coronary arterial pressure (CAP) sinusoidally around one of four mean CAP levels with a constant amplitude at several frequencies in isolated, isovolumically contracting canine heart preparations. From the frequency spectrum of the peak left ventricular pressure (PLVP) response, we arrived at a phenomenological model with two components coupled in parallel and estimated the parameter values. One component is a first-order delay system having a short time constant (approximately 1 s) and a small gain (range 0.08-0.16); it probably represents a hydraulic effect of coronary perfusion. The other component is a second-order delay system with longer time constants (approximately 15 and 6 s) and a larger gain (range 0.14-0.69); this probably represents a metabolic effect. The gain value of the slow component varied inversely with the mean CAP level, and studies with adenosine suggested that this dependence was due to the coronary autoregulation. The model prediction of the transient responses of PLVP to step changes in CAP agreed reasonably well with those experimental data of transient responses obtained in the identical hearts but not used to determine the model parameter values.

A 63-year-old man with a history of Pustulosis Palmaris et Plantaris since 1960 was suffered from lumbago and anterior chest pain in 1973. Chest X-ray film showed inter-sterno-costo-clavicular ossification. Spondylodiscitis, Syndesmophyte of lumbal spine, Sacro-iliitis were seen by abdominal X-ray film. He was admitted to near hospital with generalized edema and urinary protein (10-20 g/day) in April 1986. Laboratory studies showed hypoproteinemia, positive C-reactive protein, high serum amyloid A protein(240 ng/100 microliters) and negative HL-A B27. He was diagnosed Pustulotic arthro-osteitis and Secondary Amyloidosis by renal biopsy and rectal biopsy in 1987. He was treated with high-dose oral prednisolone(60 mg/day)and dimethyl sulfoxide. After treatment, Edema and Nephrose improved. Increasement of renal function was not observed.

The immunological and virological status of three hemophiliacs infected with human immunodeficiency virus type 1 (HIV-1) was monitored for 11 months. One of these patients was also infected with human T cell lymphotropic virus type 1 (HTLV-1), and HIV-1 could be isolated only from this patient among the three subjects. The doubly infected subject had the fewest T4 (helper) lymphocytes and the highest proportion of T8 lymphocytes with DR human leukocyte antigens (DR-Ag). The serum level of HIV-1 antigen increased in this patient during the observation period, and this increase was accompanied by a decrease in the proportion of DR-Ag-positive cells among the T8 lymphocytes. This patient was treated with 800 mg of zidovudine daily for 50 days. With treatment, the nonspecific clinical symptoms improved and the proportions of DR-Ag positive cells among the T8 lymphocytes decreased. Serum levels of HIV-1 antigen decreased immediately when therapy was started but later increased during therapy. In persons infected with both HTLV-1 and HIV-1, HIV-1 seems to proliferate readily.

The effects of mechanical changes in loading conditions on the left ventricular end-systolic pressure-volume relation (ESPVR) were studied in nine open-chest dogs, including three dogs studied before and after beta-adrenergic blockade. Left ventricular pressure was measured with a micromanometer, and left ventricular volume was measured with a conductance catheter. ESPVRs were obtained by increasing left atrial inflow over wide volume ranges (as much as threefold) under three different conditions: control or high or low aortic impedance. High impedance was obtained by occlusion of the descending aorta, and low impedance was obtained by a shunt between the subclavian artery and the left atrium. In the unblocked animals in 21 of 28 runs, a second-order polynomial equation gave a better fit for the ESPVR than a linear relation. To quantify the effects of the changes in aortic impedance on the ESPVR, we calculated from the quadratic equation its volume intercept (V18) and its local slope (E18) at an end-systolic pressure (Pes) of 18 kPa. In the unblocked animals, a statistically significant difference was found in V18 between low impedance (21.50 +/- 6.27 ml) and high impedance (14.10 +/- 8.98 ml; p less than 0.005) and between control (19.14 +/- 9.58 ml) and high impedance (p less than 0.05). In most dogs, E18 was increased at high and decreased at low impedance, but not significantly. In the additional experiments with beta-blockade, the nonlinearity diminished somewhat, but the load dependency of the ESPVR remained present after beta-blockade because the same leftward shift of the ESPVR with high aortic impedance was found. Two other relations, namely, of dP/dtmax and of stroke work versus end-diastolic volume, were also investigated, which on the whole showed the same behavior as the ESPVR. These results indicate that the ESPVR and dP/dtmax-Ved and stroke work-end-diastolic volume relations, when studied over a wide volume range, are nonlinear and that changes in loading conditions influence indexes of contractility derived from these relations, especially the volume intercepts, in such a way that an increase in aortic impedance may be interpreted as an increase in contractility. Blocking the beta-adrenergic receptors did not influence the load dependency of the ESPVR but, in some cases, tended to decrease the nonlinearity in concordance with the relation between contractility and nonlinearity in isolated hearts.

A patient with chronic T-cell leukemia characterized by a suppressor phenotype is reported. A 71-year-old woman presented with symptoms and signs of hypercalcemia. Peripheral blood specimen showed abnormal lymphoid cells with an oval to cleaved nucleus, rather condensed chromatin, occasional prominent nucleolus, and basophilic cytoplasms with vacuoles which seems to be a T-cell counterpart of B-cell chronic lymphocytic leukemia with mixed cell types. The phenotype of these cells was CD4-, CD8+, CD5+, CD6+ with poor expression of CD3, CD7, and CD25. Southern blot analysis of T-cell receptor beta-chain gene revealed one allele rearranged band. The serum antibodies were positive against human T-cell leukemia virus, type I-associated antigens, but monoclonal integration of proviral DNA was not detected in the leukemic cells suggesting that she was just a carrier of this virus. Interestingly, serum PTH-related peptide (PRP) was elevated. The combination therapy with vincristine and prednisolone for leukemia decreased not only the number of leukemic cells but also the serum PRP levels. The clinical course was aggressive. She only responded transiently to treatments, and died of renal failure due to uncontrollable hypercalcemia six weeks after admission.

Using KOLT-2 monoclonal antibody (mAb) recognizing a 44,000 molecular weight (MW) T-cell activation antigen (CD28), we observed the co-expression of KOLT-2 (CD28) antigen and Tac (CD25) antigen, associated with a 55,000 MW chain of the interleukin-2 receptor (IL-2R) complex. IL-2R (p55), on several T-cell lines transformed by human T-lymphotropic virus-I (HTLV-I), as well as several subclones of the natural killer (NK)-like cell line YT. When YT cells (YTC3T, YT5.1) and IL-2-dependent HTLV-I+ T-cell line cells (ED, ATL35C) expressing both IL-2R (p55) and IL-2R (p70) chains were cultured with recombinant IL-2 (rIL-2), KOLT-2 antigen was down-regulated in 24 hr. However, KOLT-2 antigen on HPB-ALL (Kurume) cells expressing neither of the IL-2R chains was unaffected by IL-2. IL-2 also failed to down-regulate KOLT-2 antigen on MT-1 cells bearing IL-2R (p55)/Tac without IL-2R (p70). To clarify the role of IL-2R (p70) in the IL-2-induced down-regulation of KOLT-2 antigen, we analysed the effect of IL-2 on a Tac- subclone of YT (YT2C2) that expresses IL-2R (p70). As was the case with Tac+ YT cells, KOLT-2 antigen was down-regulated by IL-2, and this down-regulation was inhibited by anti-IL-2R (p70) mAb but not by anti-Tac mAb. These results show that the signalling through the IL-2/IL-2R system down-regulates KOLT-2 (CD28) antigen by way of the interaction between IL-2 and IL-2R (p70), irrespective of IL-2R (p55)/Tac. Possible involvement of the IL-2/IL-2R system in the CD28-mediated mitogenic mechanism is discussed.

Epinephrine promotes resuscitation from ventricular fibrillation because of its peripheral vasoconstrictive effects. However, the beta-adrenergic effects of epinephrine may be detrimental because of the stimulation of myocardial oxygen demand. To test whether functional recovery from fibrillation in hearts treated with a selective alpha-adrenergic agent is greater than in hearts treated with epinephrine, ventricular fibrillation was induced in eight isolated dog hearts while coronary perfusion pressure was maintained at 30 mm Hg. In random order, epinephrine (5 micrograms/min), phenylephrine (50 micrograms/min) or no drug was infused for 5 min. The heart was then defibrillated, the drug infusion stopped and coronary perfusion pressure increased to 100 mm Hg. Coronary blood flow (ml/min per 100 g), arteriovenous oxygen difference (ml O2/dl) and myocardial oxygen consumption (ml O2/min per 100 g) measured after 4 min of ventricular fibrillation were greater with epinephrine (mean +/- SD 30.9 +/- 11.7, 17.5 +/- 1.6 and 5.4 +/- 1.9, respectively) than with phenylephrine (24.4 +/- 6.0, 15.7 +/- 2.6 and 3.8 +/- 1.1, respectively) or no drug (19.8 +/- 5.2, 12.8 +/- 1.8 and 2.6 +/- 0.7, respectively) (p less than 0.05, p less than 0.05 and p less than 0.05, respectively). The slope of the end-systolic pressure-volume relation 10 min after defibrillation and restoration of normal coronary perfusion pressure was depressed (percent of prefibrillation value) most by epinephrine infusion (72 +/- 17%, n = 6), less by no drug infusion (82 +/- 12%, n = 4) and was increased after phenylephrine infusion (143 +/- 17%, n = 6) (p less than 0.002).(ABSTRACT TRUNCATED AT 250 WORDS)

Regional cerebral blood flow (rCBF) was determined by the 133Xe inhalation technique (Headtome II: ring detection SPECT) in 53 DSM-III schizophrenic patients. The rCBF values were corrected by using end-tidal carbon dioxide concentration values (PECO2). After rCBF measurement, neuropsychological tests--Word Fluency Test, Maze Test and Wisconsin Card Sorting Test--were performed. There were significant correlations between frontal rCBF and scores on each neuropsychological test. In particular, a moderate correlations between the frontal rCBF and the performance on the Wisconsin Card Sorting Test was noted. It seems likely that decrease of rCBF in prefrontal regions at rest reflects a disturbance of frontal lobe function in schizophrenic patients.

We tested the afterload and contractile state dependency of three indexes of myocardial oxygen consumption (MVO2): total energy requirement (Et), pressure work index (PWI), and pressure-volume area (PVA). MVO2 was measured in seven isolated canine hearts at four or five different end-diastolic volumes at each of three settings of afterload resistance and with the hearts contracting isovolumically. In several hearts, contractility was also varied by dobutamine infusion. Measured MVO2 (MMVO2) was compared with values predicted (PMVO2) by each index. There was always a high degree of correlation between MMVO2 and PMVO2 for each of the indexes. However, there was a large degree of variability in the coefficients of the MMVO2-PMVO2 relation from one heart to another. We also observed a statistically significant influence of both afterload and contractile state on the predictive power of each of the indexes. Thus each index that we tested had shortcomings in being able to predict MVO2 accurately over a wide range of hemodynamic conditions.

According to the three hypotheses on the regional brain dysfunction in schizophrenia that have received some support in studies of cerebral blood flow (CBF) and cerebral metabolic rate, we calculated eight CBF measurement indices in 59 schizophrenic patients; frontality, laterality, cortical to subcortical gradients and superior to inferior difference. Four factors were selected from these eight indices, treated by principal component factor analysis (factor 1: cortical to subcortical gradient; factor 2: inferior frontality; factor 3: superior frontality; factor 4: laterality). We investigated their correlations with clinical and demographic characteristics. Factor 1 correlated with duration of illness. Factor 2 related most highly to numbers of perseverative errors on the Wisconsin Card Sorting Test and moderately to anhedonia. Factor 4 related to attentional impairment score of the Scale for the Assessment of Negative Symptoms. The schizophrenia specific symptom score calculated from the Brief Psychiatric Rating Scale did not relate to any of these factors. It seemed that there were various dimensions of neural deficits in schizophrenia, corresponding to various aspects of symptomatology or neuropsychological functions.

To study the end-systolic pressure-volume relationship of left ventricle ejection against physiological afterload, we imposed seven simulated arterial impedances on excised canine left ventricles connected to a newly developed servo-pump system. We set each of the impedance parameters (resistance, capacitance, and characteristic impedance) to 50, 100, and 200% of normal value (resistance: 3 mm Hg sec/ml; capacitance: 0.4 ml/mm Hg; characteristic impedance: 0.2 mm Hg sec/ml), while leaving the other parameters normal. Under a given impedance, the end-systolic pressure-volume relationship was determined by preloading the ventricle at four different end-diastolic volumes. There was no significant change in the slope of the end-systolic pressure-volume relationship with changes in any of the afterloading impedance parameters. However, the volume intercept of the end-systolic pressure-volume relationship decreased significantly with resistance from 5.5 +/- 1.0 (SE) ml at resistance equal to 1.5 mm Hg sec/ml to 0.6 +/- 1.8 ml at resistance equal to 6 mm Hg sec/ml (P less than 0.01). The volume axis intercept also decreased with characteristic impedance, from 5.9 +/- 2.0 ml at a characteristic impedance of 0.1 mm Hg sec/ml to 5.4 +/- 2.1 ml at a characteristic impedance of 0.4 mm Hg sec/ml, (P less than 0.05). We conclude that the slope of the end-systolic pressure-volume relationship is insensitive to a wide range of changes in afterload impedance, but its volume intercept is dependent on resistance and characteristic impedance.

We developed a framework of analysis to predict the stroke volume (SV) resulting from the complex mechanical interaction between the ventricle and its arterial system. In this analysis, we characterized both the left ventricle and the arterial system by their end systolic pressure (Ps)-SV relationships and predicted SV from the intersection of the two relationship lines. The final output of the analysis was a formula that gives the SV for a given preload as a function of the ventricular properties (Ees, V0, and ejection time) and the arterial impedance properties (modeled in terms of a 3-element Windkessel). To test the validity of this framework for analyzing the ventriculoarterial interaction, we first determined the ventricular properties under a specific set of control arterial impedance conditions. With the ventricular properties thus obtained, we used the analytical formula to predict SVs under various combinations of noncontrol arterial impedance conditions and four preloads. The predicted SVs were compared with those measured while actually imposing the identical set of arterial impedance conditions and preload in eight isolated canine ventricles. The predicted SV was highly correlated (P less than 0.0001) with the measured one in all ventricles. The average correlation coefficient was 0.985 +/- 0.004 (SE), the slope 1.00 +/- 0.04, and the gamma-axis intercept 1.0 +/- 0.2 ml, indicating the accuracy of the prediction. We conclude that the representations of ventricle and arterial system by their Ps-SV relationships are useful in understanding how these two systems determine SV when they are coupled and interact.