TNF-alpha induces protein synthesis through PI3-kinase-Akt/PKB pathway in cardiac myocytes.

The activation of phosphatidylinositol (PI) 3-kinase and Akt/protein kinase B (PKB) by tumor necrosis factor (TNF)-alpha and their roles on stimulation of protein synthesis were investigated in cultured neonatal rat cardiac myocytes. Treatment of cells with TNF-alpha resulted in enlargement of cell surface area and stimulation of protein synthesis without affecting myocyte viability. TNF-alpha induced marked activation of PI3-kinase and Akt/PKB, and the activation of PI3-kinase and Akt/PKB was rapid (maximal at 10 and 15 min, respectively) and concentration dependent. Akt/PKB activation by TNF-alpha was inhibited by a PI3-kinase-specific inhibitor LY-294002 and adenovirus-mediated expression of a dominant negative mutant of PI3-kinase, indicating that TNF-alpha activates Akt/PKB through PI3-kinase activation. Furthermore, TNF-alpha-induced protein synthesis was inhibited by pretreatment with LY-294002 and expression of a dominant negative mutant of PI3-kinase or Akt/PKB. These results indicate that activation of the PI3-kinase-Akt/PKB pathway plays an essential role in protein synthesis induced by TNF-alpha in cardiac myocytes.

A case of pulmonary acariasis in lung of Japanese macaque.

A 20-year-old female Japanese macaque, weighing 8.7 kg, developed severe pulmonary acariasis. Numerous whitish nodules, 2-4 mm, were scattered throughout the lungs. Histologically multifocal granulomatous lesions consisting of a large number of eosinophils, epithelioid cells, foreign body type giant cells, and collagen fibers were aggregated around the mait bodies. Numerous mast cells were also detected in the lesions by toluidine blue staining, and tested positive for tryptase by immunohistochemistry. This may be the first reported case of severe pulmonary acariasis in a Japanese macaque.

Measurement of urinary amino acids using high-performance liquid chromatography equipped with a strong cation exchange resin pre-column.

We determined the urinary amino acid concentrations by high-performance liquid chromatography (HPLC) using an eluent buffer which had been processed through a column packed with strong cation-exchange resin. With this column, ghost peaks resolved or much reduced in size and shifted to a post-arginine position. A clearer separation of peaks of urinary amino acids and ammonia was obtained. This could be due to the elimination of O-phthalaldehyde reactive substances in the eluent by the resin column. This method does not require a separate step to remove ammonia, unlike most of the methods currently used, and thus saves time.

Activated RhoA stimulates c-fos gene expression in myocardial cells.

Rho regulates various cell functions, including cell morphology and motility. However, the functional role of Rho on the signaling pathway in myocardial cells (MCs) is unknown. In the present study, we attempted to explore the mode of Rho action for c-fos gene expression in MCs. Expression of the c-fos promoter/enhancer linked to the luciferase reporter gene (c-fos luciferase) was stimulated by the wild type of RhoA and the point-mutated active form of RhoA (RhoA Val14) but not the biologically inactive effector domain mutant of RhoA. Rho GDP dissociation inhibitor inhibited the action of RhoA on c-fos luciferase expression. The deletion analysis revealed that the c-fos serum response element (SRE) and the 12-O-tetradecanoylphorbol-13-acetate response element (TRE) mainly account for c-fos luciferase expression by RhoA Val14. The c-fos SRE mutant, which contains an intact binding site for the serum response factor but lacks the ternary complex factor binding site, was activated by RhoA Val14. The action of RhoA Val14 on c-fos luciferase expression was not inhibited by downregulation of protein kinase C, protein kinase C inhibitors, or tyrosine kinase inhibitors. These results indicate that activated RhoA stimulates c-fos gene expression through the c-fos SRE and TRE and that the signaling pathway from activated RhoA to the c-fos promoter/enhancer is independent of these inhibitor-sensitive pathways in MCs.

Systemic capillary leak syndrome associated with multiple myeloma of IgG kappa type.

A case of systemic capillary leak syndrome in a 67-year-old woman is reported. The patient suddenly presented with severe edema and hypotension associated with an increased hematocrit and a decreased level of serum albumin, suggesting capillary hyperpermeability. The patient had IgG kappa monoclonal gammopathy in association with not only a slight increase in plasma cells in the bone marrow but also several punched out lesions in the skull, indicating multiple myeloma. Although most of the cases of systemic capillary leak syndrome reported previously were accompanied by monoclonal gammopathy, this appears to be the first case in which capillary leak syndrome developed in a patient with multiple myeloma.

The role of the spleen in endotoxin-induced liver injury.

In these experiments, the role of the spleen in endotoxin-induced liver injury was evaluated, using rats which underwent splenectomy or splenic vein ligation with antecedent spleno-systemic shunt. Male Wistar rats were divided into three groups: a sham-operated group, a splenectomy group, and a splenic vein ligation group. In each animal, 48 h after surgery, 5 mg/kg lipopolysaccharide (LPS) were injected intravenously. Six rats from each group were sacrificed 6 or 12 h after LPS administration. Bronchoalveolar lavage fluid (BALF) and arterial blood were also collected. Splenectomy reduced the liver injury as indicated by the serum lactate dehydrogenase level. A decrease in liver tissue adenosine triphosphate and increase in lipid peroxide were induced by LPS administration and inhibited by splenectomy. Splenectomy also reduced alveolar protein release as indicated by the protein level in BALF. Splenic vein ligation provided similar protective effects on the liver, but did not affect lung injury. From these results, it appears that the spleen plays a significant role in endotoxin-induced liver injury, and a mediator derived from the spleen is likely associated with development of liver injury. This mediator may be cleared or inactivated by not only splenectomy but also splenic vein ligation.

Effects of hepatic arterial infusion chemotherapy on unresectable or recurrent hepatocellular carcinoma.

We performed hepatic arterial infusion chemotherapy (HAI) on 86 patients with unresectable hepatocellular carcinoma (HCC, 61 patients) or unresectable recurrent HCC after hepatectomy (25 patients). As drug therapy, 250 mg of 5-fluorouracil was injected daily for 14 days using a reservoir embedded in the subcutaneous layer. During this period, 0.4 mg/kg of doxorubicin and 0.12 mg/kg of mitomycin C suspended in Lipiodol Ultra-Fluide were also injected twice intra-arterially. This was defined as one course of HAI, and it was repeated every 3 months. In the patients with unresectable HCC, the 1-, 2-, and 3-year survival rates were 31.5%, 22.4%, and 10.7%, respectively, and the numbers of cases showing a complete response (CR), a partial response (PR), a minor response (MR), no change (NC), and progressive disease (PD) according to the Criteria for the Evaluation of the Clinical Effects of Solid Cancer Chemotherapy established by the Japan Society for Cancer Therapy were 1 (1.6%), 20 (32.8%), 5 (8.2%), 28 (45.9%), and 7 (11.5%), respectively. On the other hand, the 1-, 2-, and 3-year survival rates of the patients with unresectable recurrent HCC were 69.6%, 34.8%, and 14.9%, respectively. The rate of catheter patency after 1 year was 64.1%, and the mean catheter-patency period was 311.9 days. Patients in group A (CR+PR, n = 21) survived significantly longer than those in group B (MR+NC+PD, n = 40; P < 0.05). In conclusion, since responders to HAI achieve longer survival than nonresponders, the selection of effective drugs is important for this therapy.

Serum levels of coenzyme Q10 and lipids in patients during total parenteral nutrition.

Serum levels of coenzyme Q10 (CoQ10) as well as lipids were determined in patients during total parenteral nutrition (TPN). The mean CoQ10 levels (M +/- SD) were 0.77 +/- 0.30 microgram/ml for 108 normal subjects and 0.59 +/- 0.35 microgram/ml for 95 patients before TPN. The mean CoQ10 level of the patients decreased significantly to 0.35 +/- 0.23 microgram/ml one week after the start of TPN, and then remained almost unchanged during TPN for up to 6 weeks. When the patients receiving TPN (TPN patients) were grouped according to their clinical diagnoses, the mean CoQ10 level of patients with cancer was significantly lower than that of the other patients without cancer in 4 week therapy, but there was no difference in the levels between the patients with and without diseases of the gastrointestinal tract. Serum levels of total cholesterol (T-Chol) and esterified cholesterol in TPN patients also declined below their respective normal ranges, but not to the same extent in comparison to CoQ10. The levels of triglycerides (TG), phospholipids (PL), non-esterified fatty acids, low density lipoproteins, very low density lipoproteins, chylomicrons, and cholesterol in the high density lipoprotein fraction in serum of TPN patients were within their normal ranges. The levels of CoQ10 in TPN patients were correlative to those of T-Chol, TG, and PL, and decreased rapidly prior to the latter levels.

High-performance liquid chromatography of coenzyme Q-related compounds and its application to biological materials.

A convenient and precise method for the separation and determination of coenzyme Q (CoQ)-related compounds (CoQ homologues, plastoquinone-9, ubichromenol-9, etc.) was developed using high-performance liquid chromatography (HPLC). All compounds tested were separated using a reverse-phase column with a suitable mobile phase and detected at a wavelength of 275 nm. CoQ extracts in plasma and erythrocytes were purified by thin-layer chromatography prior to HPLC analysis, but such purification was not necessary when determining CoQ in urine and tissues. Hydroquinone forms of CoQ existing in animal tissues were oxidized to the corresponding quinone forms with potassium hexacyanoferrate(III). This HPLC method was applied satisfactorily to the determination of the contents of CoQ homologues in human and animal samples. CoQ10 was the only homologue detected in human samples, and CoQ8, CoQ9 and CoQ10 were native homologues of CoQ in rat tissues. Ubichromenol-9 and plastoquinone-9 were not detected in these samples.

[Inspection of inner structure of sealed 60Co sources by the autoradiography with radcolor film].

In studies using 60Co gamma-ray irradiation, an extremely high dose rate is often required and samples must be placed very near a 60Co source. The dose rate in the vicinity of the source depends on the inner structure of the source. Therefore, it is necessary to examine the distribution of source activity. As a method of the examination an autoradiographic technique using several coloring materials for irradiation was tried. The materials used were "Radcolor film" developed by our center, besides a glass, polymethylmethacrylate (PMMA) and a blue cellophane. Results showed that the autoradiography with Radcolor film is very useful to examine the position, the size and the activity difference of 60Co pieces by the change of color.

[Gel-type agar color dosimeter].

Gel-type color dosimeter which is made by dissolving 2 kinds of dye in agar is developed. The dosimeter has following characteristics. (1) The dosimeter is easy to make and can be formed easily to any shape required. (2) The dose and dose distribution in krad region can be measured by color change of the dosimeter. (3) The 3 dimensional dose distribution can be observed by cutting or slicing the irradiated dosimeter. The dosimeter is used in the sprout inhibition experiment of potato by electron irradiation. From the data obtained, several irradiation conditions of the irradiating machine and irradiation method can be decided most suitably.

Vitamin A and E requirements during total parenteral nutrition.

Studies were undertaken to determine rational dosages of vitamin A and E during long-term total parenteral nutrition (TPN). Four kinds of vitamin prescriptions containing different amounts of vitamin A and E were prepared from commercially available products and/or hospital pharmacy products. Patients were divided into four groups according to the vitamin prescription used. Plasma vitamin levels of different patient groups were determined by a modified fluorimetric method and were compared with those of a normal subject group. The stability of vitamin A and E in TPN solution after admixing was determining by measuring the remaining vitamin contents by high pressure liquid chromatography. From the results, it was concluded that 1) about 50% of vitamin A was decomposed by sunlight (about 2000 lux) 3 hr after admixing and an orange-colored vinyl cover could protect its photodecomposition; 2) vitamin E was stable at any condition tested; 3) 2500 IU of vitamin A and 15 IU of vitamin E could meet the daily requirements; 4) the plasma levels of vitamin A and E were correlative (p less than 0.01); and 5) concomitant administration of vitamin E was essential to keep the poorer level of vitamin A in plasma.

Thiamin and pyridoxine requirements during intravenous hyperalimentation.

Studies were undertaken to determine rational dosages of vitamin B1 and B6 during long-term intravenous hyperalimentation, using more sensitive techniques than formerly used to evaluate B1 and B6 status. A standard vitamin combination, type A, (usually commercially available products) has been used up to now because of convenience, disregarding the effects of long-term administration. This combination lacks biotin, folic acid, and vitamin E and contains from 10 to 100 times the dietary allowances of such vitamins as B1, B2, B6, B12, and C. In response to the possibility of vitamin overdose, two new vitamin combinations, type B (from commercial products) and type C (a convenient and easily administered combination produced at the hospital) were developed in order to provide the normal dietary allowances and at the same time eliminate any harmful side-effects. From the results obtained, 5 mg/day for thiamin HCl and 3 mg/day for pyridoxine HCl in type B and type C were found to be a sufficient and safe level as opposed to 55 mg/day for thiamin HCl and 102 mg/day for pyridoxine HCl in type A.