Granule-dependent killing of Toxoplasma gondii by CD8+ T cells

Immunization of mice with live bradyzoites of a low-virulent Beverley strain of Toxoplasma gondii has been shown to increase CD8+ T-cell mediated immunity against a highly virulent RH strain. We found that preimmunization with an RH homogenate further enhanced this immunity. Using this model, we investigated the mechanism of CD8+ T-cell mediated protection against T. gondii infection. Splenic cells from mice immunized with RH homogenate and live bradyzoites stimulated apoptosis of RH-infected J774A.1 macrophages in vitro, and at the same time, the immunization significantly suppressed the proliferation of parasites within macrophages, as assessed by measuring 3H-uracil uptake by the parasites. Splenic cells from the immunized mice produced larger amounts of interferon-gamma (IFN-gamma) than did naive splenic cells; however, the production of nitric oxide (NO) by RH-infected macrophages was not enhanced. The elimination of CD8+ T cells from splenic cells significantly reduced their inhibitory action on parasite proliferation as well as their cytotoxic activity against RH-infected macrophages, but it did not affect the production of IFN-gamma. Treatment of CD8+ T-enriched splenic cells from the immunized mice with concanamycin A, but not an anti-Fas ligand monoclonal antibody, significantly reduced their anti-proliferative and killing capabilities, suggesting that the CD8+ T cells induced by immunization with RH antigen and live bradyzoites of the Beverley strain may exert protection against T. gondii infection at least in part through granule-dependent cytotoxic activities.     

Accumulation of p53 in esophageal squamous cell carcinoma

D-mannoheptulose is a specific inhibitor of D-glucose phosphorylation by hexokinase isoenzymes. In the present study, the phosphorylation of this heptose was investigated by either a spectrophotometric or radioisotopic procedure. Using yeast hexokinase, the phosphorylation of 25 mM D-mannoheptulose only represented 0.02% of that of 5 mM D-glucose. Such a percentage was increased to 3.93% in the case of bovine heart hexokinase. In the latter case, the Km for D-mannoheptulose was close to 0.2 mM and both D-glucose (0.1-1.0 mM) and D-glucose 6-phosphate (also 0.1-1.0 mM) inhibited the phosphorylation of the heptose (0.03-0.60 mM). Human B-cell glucokinase also catalyzed the phosphorylation of D-mannoheptulose (0.1 mM), which was now increased in a bell-shaped manner by D-glucose (1.0-20 mM). Likewise, rat parotid gland, liver and pancreatic islet homogenates catalyzed the phosphorylation of D-[3H]mannoheptulose. The results obtained in these three tissues differed from one another by their absolute values (per mg wet wt.), relative values (by reference to the phosphorylation rate of 10 mM D-glucose), and sensitivity to inhibition by D-glucose (10 mM).     

Cation effects on the voltammetric behavior of α-Keggin-type [SiMo12O40]4− and [PMo12O40]3− complexes in CH3COCH3 and CH3CN

The effect of small cations such as H⁺, Li⁺ and Na⁺ on the voltammetric behavior of α-Keggin-type [SiMo₁₂O₄₀]^4? and [PMo₁₂O₄₀]^3? complexes was investigated in CH₃COCH₃ and CH₃CN. For the [SiMo₁₂O₄₀]^4? complex, the presence of Li⁺ or Na⁺ caused the one-electron waves to be converted into a two-electron wave at ca. 0.3 V more positive than the first one-electron wave. In the presence of Li⁺ or Na⁺, the [PMo₁₂O₄₀]^3? complex underwent a two-electron reduction at the same potential as the original first one-electron wave in CH₃COCH₃, whereas it exhibited only successive one-electron waves in CH₃CN. The addition of a trace amount of H⁺ produced new two-electron waves at more positive potentials. These findings give a clue to the understanding of the reactivity of polyoxometalates as redox catalysts.     

Rationale for the use of combination angiotensin-converting enzyme inhibitor and angiotensin II receptor blocker therapy in heart failure.

BACKGROUND: The present multicenter study investigated whether the combination of angiotensin-converting enzyme inhibitor (ACEI) and angiotensin II receptor blocker (ARB) is more beneficial for preventing left ventricular remodeling and suppressing neurohumoral factors than either ACEI or ARB alone. METHODS AND RESULTS: One hundred and six patients with mild-to-moderate congestive heart failure treated in 26 Japanese institutes were randomly assigned to the combination therapy or monotherapy. Changes in physical activity (New York Heart Association functional classes, Specific Activity Scale (SAS)), concentrations of neurohumoral factors (plasma renin activity, angiotensin II, aldosterone, and brain natriuretic peptide (BNP)), and cardiac function for 6 months were compared between the 2 groups. It was found that the combination therapy, which was administered at doses standard in Japan, increased the SAS score (4.5 +/- 1.5 to 4.9 +/- 1.5, p<0.05) and decreased the plasma BNP concentration (183 +/- 163 to 135 +/- 118 pg/ml, p<0.05). In contrast, there were no changes in SAS score (4.5 +/- 1.4 to 4.6 +/- 1.4, NS) or BNP concentration (156 +/- 157 to 151 +/- 185 pg/ml, NS) in the patients receiving monotherapy. CONCLUSIONS: The results of the study demonstrate that the combination therapy, even at the standard doses for Japan, improves physical activity and plasma BNP concentration more than the monotherapy. A larger study is required to assess the effects of the combination therapy on major clinical outcomes.     

A weight reduction and weight maintenance program with long-lasting improvement in left ventricular mass and blood pressure.

Obesity is a major risk factor for cardiovascular disease and is associated with hypertension and increased left ventricular mass (LVM). Maintenance of reduced weight has been a matter of recent concerns in the treatment of obese subjects. This study was conducted to confirm the effect of the addition of exercise to diet on maintenance of body weight in a weight reduction program. In addition, this study was conducted to estimate whether LVM changes in parallel with a change in body weight during a long-term follow-up after a weight-reduction program. Twenty-two normotensive (NT) obese subjects and 14 mild hypertensive (HT) obese subjects ranging in age from 22 to 51 years participated in a 12-week supervised weight-reduction program involving mild exercise and a mild hypocaloric diet. After this 12-week intervention, they were advised to maintain their modified lifestyle during a 1-year follow-up period. After the 12-week intervention, the mean reductions in body weight (BW) in the NT and HT groups were 4.1 kg (P < .0001) and 5.8 kg (P < .0001), respectively. LVM in the NT and HT groups was significantly reduced from 154 g to 136 g (P < .005) and from 169 g to 152 g (P < .002), respectively. One year after intervention, the mean gains in BW in the NT and HT groups were 2.3 kg (not significant, NS) and 0.4 kg (NS), respectively. The mean gains in LVM in the NT and HT groups were 8 g (NS) and 7 g (NS), respectively. It was also shown that blood pressures in the HT group were significantly decreased after the 12-week intervention and there was no significant change in blood pressure in the HT group 1 year after intervention. In conclusion, reduced body weight was maintained for 1 year after a 12-week supervised weight-reduction program in both normotensive and mild hypertensive obese subjects. Reduced left ventricular mass was maintained for a long period in both normotensive and mild hypertensive obese subjects and lowered blood pressure was maintained in the mild hypertensive obese subjects.     

Human V delta 2+ gamma delta T-cell tolerance to foreign antigens of Toxoplasma gondii.

Little is known about the mechanisms involved in human gammadelta T-cell tolerance to self or to foreign antigens. Patients with congenital toxoplasmosis offer a unique opportunity to examine Vdelta2+ gammadelta T-cell tolerance. Analysis of gammadelta T cells in patients with congenital toxoplasmosis revealed evidence for anergy of these cells with or without clonal Vdelta2+ gammadelta T-cell expansion in the acute phase of the Toxoplasma infection. T cells in general were unresponsive and did not proliferate upon exposure to mitogens or to Toxoplasma lysate antigens or in response to live Toxoplasma-infected cells when the congenitally infected infants were 1 month of age, and they exhibited selective anergy to Toxoplasma lysate antigens and live Toxoplasma-infected cells when the infants were aged 5 months. During the chronic phase of congenital toxoplasmosis in the patients who were more than I year of age, the repertoires of the gammadelta T-cell receptors were found to be within normal ranges. In addition, in the chronic phase, the gammadelta T cells proliferated and secreted gamma-interferon in response to exposure to live Toxoplasmia-infected cells. By contrast, alphabeta T cells remained anergic. Vdelta2+ gammadelta T cells have been considered to undergo extrathymic maturation and thus to be subject to development of peripheral tolerance. Our findings indicate that Vdelta2+ gammadelta T-cell tolerance was lost in these infected infants earlier than alphabeta T-cell tolerance. These findings suggest that gammadelta T cells play a role in protection against Toxoplasma gondii in the chronic phase when congenitally infected children are more than 1 year of age, especially in those in whom alphabeta T cells continue to exhibit deficits in specific immune responses to Toxoplasma antigens.     

Marked secretion of pyruvate in human duodenal juice stimulated with pancreozymin or secretin

Pyruvate and lactate in duodenal aspirates were investigated to determine whether they are excreted from human pancreas as substrates for alkaline secretion as is bicarbonate. Secretion of these acids was compared with that of another organic acid, citrate, which is thought to be excreted in close relationship to digestive enzymes. All acids were assayed in the fluid obtained from 11 subjects without pancreatic diseases, before and after sequential intravenous injections of 1 unit/kg pancreozymin and 1 unit/kg secretin. Pyruvate concentrations were markedly increased by each stimulation, especially by secretin, and the cumulative excretions of pyruvate and bicarbonate after secretin stimulation were significantly correlated among the subjects. In contrast, lactate concentrations, although high just after administration of pancreozymin, declined to a considerable extent following each injection, rather similar to those of protein or citrate. These data suggest that pyruvate may be secreted from human pancreatic duct cells similar to bicarbonate secretion through mechanisms related to alkaline secretion.     

Expression of oncogenes in human liver disease

To elucidate the role of oncogene expression in hepatocarcinogenesis, we examined the expression of 8 cellular oncogenes by dot blot and/or northern blot analysis in neoplastic, cirrhotic and non-cirrhotic human liver tissues obtained at surgery. Significantly higher levels of c-myc gene expression were observed in tissues of hepatocellular carcinoma (HCC) and adjacent cirrhotic tissues than in apparently normal liver tissues or those of chronic hepatitis (normal-chronic hepatitis). There was a tendency to higher c-myc mRNA levels in HCC than in liver cirrhosis. However, when tumorous and adjacent cirrhotic tissues from the same patient were compared, c-myc mRNA levels were not consistently higher in HCC. No significant differences in mRNA levels of c-fos, N-myc, N-ras, Ha-ras, c-erbA, c-erbB and c-abl were observed among the HCC, cirrhosis and normal-chronic hepatitis groups. Although the significance of increased c-myc gene expression in liver cirrhosis and HCC is still not known, it is conceivable that the persistent elevation of c-myc gene expression in cirrhosis contributes to the development of HCC.     

Marrow transplantation from tolerant donors to treat and prevent autoimmune diseases in BXSB mice.

Autoimmune-prone BXSB male mice were supralethally irradiated and transplanted with CBA/H bone marrow cells. A complete and long-term chimerism was established when donor mice had been induced to develop tolerance of BXSB male antigens by combined treatment with BXSB male spleen cells and cyclophosphamide. Such chimeras did not express autoimmune phenomena or develop lethal autoimmune manifestations. Nor did the recipient mice develop the wasting syndrome or evidence of persistent immunodeficiencies that have been seen in other strains of autoimmune-resistant mice that had been transplanted with bone marrow cells across major histocompatibility complex barriers following an initial purging of the bone marrow of Thy-1+ cells using anti-Thy-1+C.