Inhibition by gossypol of tumor promoter-induced arachidonic acid metabolism in rat peritoneal macrophages.

Rat peritoneal macrophages were prelabeled with [3H]arachidonic acid. The release of radioactivity into the medium was increased by treatment with TPA-type tumor promoters, such as TPA, teleocidin and aplysiatoxin, and the non-TPA-type tumor promoter, thapsigargin. Gossypol, at concentrations of 3 and 10 microM, inhibited the release of radioactivity stimulated by both types of tumor promoter, although the mechanism of stimulation of arachidonic acid metabolism is different in the two types of tumor promoter. Stimulation of prostaglandin E2 production by these tumor promoters was also inhibited by treatment with gossypol. Calcium ionophore A23187-stimulated release of radioactivity and prostaglandin E2 production were also inhibited by gossypol treatment. The mechanism of inhibition by gossypol of prostaglandin E2 production is discussed.

Dual effects of staurosporine on arachidonic acid metabolism in rat peritoneal macrophages.

Staurosporine is a microbial anti-fungal alkaloid having a most potent inhibitory activity on protein kinase C and is recently found as a non-12-O-tetradecanoylphorbol-13-acetate (non-TPA)-type tumor promoter of mouse skin, although tumor promotion induced by a TPA-type tumor promoter teleocidin is suppressed by staurosporine. When rat peritoneal macrophages were incubated in the medium containing various concentrations of staurosporine, prostaglandin E2 production and release of radioactivity from [3H]arachidonic acid-labeled macrophages were stimulated at concentrations of 1 and 10 ng/ml. But higher concentrations of staurosporine such as 100 and 1000 ng/ml showed no stimulative effect on prostaglandin E2 production although cytoplasmic free calcium levels were increased in a dose-dependent manner. Staurosporine-induced stimulation of prostaglandin E2 production was inhibited by treatment with cycloheximide, suggesting that a certain protein synthesis is prerequisite for the stimulation of arahcidonic acid metabolism. At higher concentrations (100 and 1000 ng/ml), staurosporine inhibited TPA-type tumor promoter (TPA, teleocidin and aplysiatoxin)-induced stimulation of arachidonic acid metabolism probably due to the inhibition of protein kinases. Tumor promotion activity and anti-tumor promotion activity of staurosporine might be explained by the fact that the lower concentrations of staurosporine stimulate arachidonic acid metabolism and the higher concentrations of staurosporine inhibit the tumor promoter-induced arachidonic acid metabolism, respectively.

Assessment of biological age by principal component analysis.

A method of assessing biological age by the application of principal component analysis is reported. Healthy individuals (462) randomly selected from about 6000 men who had taken a 2-day health examination were studied. Out of the 30 physiological variables examined in routine check-ups, 11 variables were selected as suitable for the assessment of biological age based on the results of factor analysis and the physiological meaning of each test. This variable set was then submitted to principal component analysis, and the 1st principal component obtained from this analysis was used as an equation for assessing one's biological age. However, the biological age calculated from this equation is expressed as a score, so the estimated score was transformed to years (biological age) using the T-score idea. The biological age estimated by this method is practically useful and theoretically valid in contrast with the multiple regression model, because this approach eliminates and overcomes the following 2 big problems of the multiple regression model: (1) the distortion of the individual biological age at the regression edges; and (2) a theoretical contradiction in that a perfect model will merely be predicting the subject's chronological age, not his biological age.

Effects of long-term, light exercise under restricted feeding on age-related changes in physiological and metabolic variables in male Wistar rats.

The effects of long-term, light exercise under restricted feeding on age-related changes in physiological and metabolic functions were examined in male Wistar rats. Adult (100 days old) rats were divided into sedentary (R10S) and exercise (R11E) groups, and given 10 and 11 g/day, respectively, of a 20% casein diet until they reached 900 days of age. Group R11E simultaneously underwent 3000 m/day of running exercise throughout the test period. As compared with the sedentary group, long-term, light exercise significantly increased body nitrogen retention and serum protein levels, decreased body fat and plasma insulin levels, prevented age-related decline in the basal metabolic rate, and reduced age-associated histopathological changes in the kidney and liver. Long-term, light exercise further enhanced the benefits of restricted feeding on age-related deterioration in physiological and metabolic variables and improved body composition, but did not prolong survival at 900 days of age.

Interaction of silkworm larval hemolymph antitrypsin and bovine trypsin.

Antitrypsin isolated from silkworm larval hemolymph can inhibit bovine trypsin. The apparent molar ratio of silkworm antitrypsin (sw-AT) to trypsin at extrapolated null trypsin activity was determined to be 1.3 by titration of trypsin with sw-AT. The undissociability of the complex between sw-AT and trypsin on sodium dodecyl sulfate-polyacrylamide gel electrophoresis was confirmed by immunoblotting and fluorescence labeling techniques. Chemical analysis of the complex elucidated that it contained equimolar sw-AT and trypsin. Densitometric analysis of electrophoretic patterns obtained during the titration of trypsin by sw-AT suggested the presence of a suicide product formed from sw-AT. This was the reason why excess sw-AT was needed for complete inhibition of trypsin. In the complex, the sw-AT molecule was cut at one site but the fragments produced were still joined together. Trypsin in the complex was released by treatment at pH 10.0, and it was deduced that the complex formation involved acyl-bond formation between sw-AT and trypsin. The sw-AT component obtained from the alkali-treated complex possessed two kinds of NH2-terminal amino acid sequences. Non-covalent forces may bind the two fragments of sw-AT, which could be separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Amino-terminal amino acid sequence analysis of the large fragment gave a sequence identical with that of intact sw-AT. This indicated that the reactive site of sw-AT with trypsin was located at the COOH-terminal region of the molecule. These characteristics resemble those of inhibitors belonging to the serpin family.

Controlled release from solid dispersion composed of poly(ethylene oxide)-Carbopol interpolymer complex with various cross-linking degrees of Carbopol.

Solid dispersion composed of the poly(ethylene oxide) (PEO)-Carbopol((R)) (CP) interpolymer complex containing phenacetin (PHE) was prepared by using six grades of CP having various cross-linking degrees. We attempted to control the medicine release from the PEO-CP solid dispersion by varying the CP grade. The powder X-ray diffraction pattern and differential scanning calorimetry curves suggested that PHE existed in the amorphous state, and PEO in the crystalline state disappeared in the solid dispersions. The release profile of PHE varied depending on the CP grade. A small release rate was observed at CP910 and CP971P that are cross-linked at low and middle degrees, respectively. The Fourier transform-infrared (FT-IR) spectra showed that the amount of the PEO-CP complex formed by hydrogen bonding changed depending on the CP grade. With the cross-linked CP, a good correlation was observed between the hydrogen bonding percent and the percent released of the PHE after 60 min (D(60 min)), indicating that PHE release was controlled by the amount of PEO-CP complex formation in the solid dispersion. These results show that it is feasible to control the medicine release from PEO-CP solid dispersion by varying the CP grade.

Control of medicine release from solid dispersion composed of the poly(ethylene oxide)-carboxyvinylpolymer interpolymer complex by varying molecular weight of poly(ethylene oxide).

Solid dispersion composed of the poly(ethylene oxide) (PEO)-carboxyvinylpolymer (CP) interpolymer complex containing phenacetin (PHE) was prepared by using nine grades of PEO having different molecular weights from 2000 to 4500000. We attempted to control the medicine release from the PEO-CP solid dispersion by varying the molecular weight of PEO. The physicochemical properties of the solid dispersion were analyzed by powder X-ray diffractometry and thermal analysis. The interaction between PEO and CP was analyzed by IR spectroscopy. Transmittance of the polymer solution was measured to study the complexation between PEO and CP. The release profile of PHE varied depending on the molecular weight of PEO. The minimum release rate was observed at the PEO molecular weight of 35000. It was found that the amount of the PEO-CP complex formation by hydrogen bonding changed depending on the molecular weight of PEO. These results indicate that it is feasible to control the medicine release from the PEO-CP solid dispersion by varying the molecular weight of PEO.

Degradation of arylsulfate by hepatic microsomes.

The enzyme liberated by some treatments and the changes in arylsulfatase C activity in chronic hepatic damage were investigated in rat liver. 1. The enzyme activity liberated by ultrasound was the highest in the conditions studied. 2. Arylsulfatase C was assayed using p-nitrophenyl sulfate in 0.25 M Tris/acetate buffer as substrate. It is shown that this method can be used to measure arylsulfatase C activity in a mixture of arylsulfatases A and B. 3. The enzyme is mainly located in the microsomal fraction in rat liver. In toxic hepatic damage, the enzyme activity decreases from the early stage; decreasing markedly in chronic hepatic damage. The activity seems to reflect damage to the microsomes and therefore arylsulfatase C activity can be a good indicator of injury to liver microsomes.

Sex-specific effects of growth hormone on hepatic 11beta-hydroxysteroid dehydrogenase activity and gene expression in hypothyroid rats.

To investigate the effects of growth hormone (GH) on 11beta-HSD1, we determined changes in hepatic 11beta-HSD1 activity in hypothyroid rats following treatment with subcutaneous (s.c) injection of GH for periods ranging from 24 h to 7 days. In male rats, hypothyroidism markedly reduced the hepatic 11beta-HSD1 activity and serum testosterone levels (p < 0.01). Subcutaneous injection of GH once daily to male hypothyroid rats for 48 h inhibited hepatic 11beta-HSD1 activity. However, the same daily dose of GH administered to male hypothyroid rats for 7 days, resulted in a marked increase in hepatic 11beta-HSD1 activity and gene expression (p < 0.01). Furthermore, daily s.c injections of GH to castrated male hypothyroid rats for 7 days reduced hepatic 11beta-HSD1 activity rather than inducing it, the same response seen in hypothyroid female rats. In addition, the treatment of castrated male hypothyroid rats with testosterone for 7 days significantly increased this enzyme activity (p < 0.01). The changes in hepatic 11beta-HSD1 were demonstrated to be associated with the testes in hypothyroid male rats following treatment with GH for 7 days. Moreover, the prolonged exposure to GH required to induce hepatic 11beta-HSD1 in intact hypothyroid male rats and the lack of a similar effect in castrated male hypothyroid rats suggests that this action is indirect and that it may be mediated by androgen production from Leydig cells of the testes and induced by the daily injections of GH. Treatment of hypothyroid male rats with GH at 6-h intervals, however, feminized the hepatic 11beta-HSD1 gene expression.

Evaluation of the ACR-NEMA standard for communications in digital radiology.

An implementation and evaluation of a prototype multivendor communications system which complies with the American College of Radiology (ACR) and the National Electrical Manufacturers Association (NEMA) standard for communications in digital radiology is discussed. The system allows communications between interfaces from different manufacturers within a networked environment. The implementation includes network software compatible with the International Standards Organization's Open Systems Interconnect standard. The experience of the implementation effort and the evaluation of the system provide the basis for a critique of the ACR-NEMA standard. It is concluded that the ACR-NEMA standard is not well suited for application to the networked environment of picture archiving and communications systems. Two possible solutions are recommended for this problem. The first is a major revision of the existing standard. The second is the development of a family of network communications standards for digital radiology.

Decreased bradykinin binding sites in fibroblasts from progressive systemic scleroderma.

The numbers of bradykinin receptors (BK-R) in cultured dermal fibroblasts from patients with progressive systemic sclerosis (PSS) and from healthy controls were measured using a receptor binding assay. The numbers of BK-R were significantly fewer in PSS fibroblasts than in control fibroblasts (P < 0.02). However, no differences in affinity were observed in BK-R between PSS and control fibroblasts. The BK-R mRNA levels were determined in PSS and control fibroblasts by Northern blot hybridization using BK-R cDNA, but no significant differences were found. These findings suggest that the decrease in BK-R in PSS fibroblasts might occur during a posttranslational step.

Tissue content of glycosaminoglycans in the diffuse idiopathic interstitial fibrosis patient.

Quantitative changes of glycosaminoglycans in lung tissues of patients with diffuse idiopathic interstitial fibrosis and patients with bronchial asthma were studied. The total amount of glycosaminoglycans in lung tissue of patients with diffuse idiopathic interstitial fibrosis increased in comparison to the lung tissue of those patients with bronchial asthma. The increase in tissue content of glycosaminoglycans was accompanied by an increase in dermatan sulfate level. The increases in total amount of glycosaminoglycans in relative proportion of dermatan sulfate were closely related to the clinical findings, including vital capacity of the lung, PaO2, serum lactate dehydrogenase, and to the pathologic pictures of the lung.

Application of acid-treated yeast cell wall (AYC) as a pharmaceutical additive I. AYC as a novel coating material.

Acid-treated yeast cell wall (AYC) was newly prepared by acidifying the cell wall of brewer's yeast and the potential to use AYC as a novel coating material was studied. AYC had an oval shape with the diameter of several microm. The rheogram of AYC aqueous dispersion showed the plastic fluid property that is generally observed in the suspension. Core tablets containing 3% of acetaminophen (AAP) were coated with the AYC aqueous dispersion containing 5% (w/v) of AYC and 0.35% (w/v) of glycerol at various coating percents. The AAP release profile from the AYC-coated tablets was studied by the JP13 paddle method using solutions at various pH. Tensile strength and permeability of oxygen and water vapor of AYC cast film were measured. The AAP release from the AYC-coated tablets showed sigmoidal release profile with an initial lag time and the duration of the lag time depended on the coating percent of AYC. The pH of the dissolution fluid or the storage at room temperature for 120 days had little affect on AAP release from the AYC-coated tablets. These results suggest that it is possible to control the start time of medicine release independent of the pH by coating of AYC, that is the time-controlled release. The AYC cast film showed a large tensile strength and an extremely small oxygen permeability coefficient and a sufficient level of water permeability coefficient in order to protect from moisture. These results present that AYC has the high utility as a novel aqueous coating material for DDS preparations.

Application of acid-treated yeast cell wall (AYC) as a pharmaceutical additive. II: effects of curing on the medicine release from AYC-coated tablets.

Acid-treated yeast cell wall (AYC) was newly prepared by acidifying brewers' yeast cell wall. Core tablets containing 3% of acetaminophen (AAP) were coated with the AYC aqueous dispersion containing 5% (w/v) of AYC and 0.35% (w/v) of glycerol. The curing of AYC-coated tablets was performed at various curing periods of time and temperatures. The effects of curing on AAP release from AYC-coated tablets, the weight and thickness of the coated layer of AYC and the water sorption into the AYC-coated tablets were studied. The tensile strength and pore size distribution of the AYC cast film were measured. In the case of 60, 80, or 100 degrees C curing, AAP release from AYC-coated tablets showed a sigmoidal release profile with an initial lag time. The duration of the lag time increased with the increasing curing time and temperature, though the release rate after the lag time hardly changed. At 120 degrees C curing, the release rate after the lag time decreased with the increasing curing time and a sustained release was observed. The weight and thickness of the AYC-coated layer and the water sorption rate into AYC-coated tablets decreased with the increasing curing time and temperature. The tensile strength of the AYC cast film increased with increasing the curing temperature, particularly at 120 degrees C curing. It is considered that the water was evaporated from the AYC-coated layer and the adhesion force between AYC particles increased during curing, making the structure of the AYC-coated layer densely firm. The changes in the duration of lag time and the release rate may be due to changes in the structure of the AYC-coated layer caused by curing. These results show that it is feasible to control the lag time and the release rate of AAP from AYC-coated tablets by varying the curing time and temperature.

Application of Carbopol to controlled release preparations I. Carbopol as a novel coating material.

We investigated the application of Carbopol(R) (CP) as a novel coating material prepared with various grades of CP having different degrees of cross-linking and molecular weights. Viscosity and spray mist size of CP aqueous solutions at various concentrations of CP were measured. Core tablets containing theophylline (TP), as a model drug, were coated with CP at various coating ratios. The TP release profile from the CP-coated tablets was studied by the JP13 paddle method. CP tablets were prepared by compressing CP powder, and the swelling behavior of the CP tablets in JP 1st fluid, purified water, and JP 2nd fluid was observed. The spray mist size of all CP aqueous solutions was small at a concentration of 1% and below, and drastically increased over a concentration of 1%. This result suggests that the appropriate concentration of the CP solution for coating is 1% or below. Sustained release of TP from the CP-coated tablets at a coating ratio of only 3% was observed in the JP 1st fluid and purified water, although fast release was observed in the JP 2nd fluid. The fast release in the latter fluid may be due to the fact that CP is an acid material. These results suggest that it is feasible to control the drug release by use of an extremely small amount of CP coating and that CP is useful as a novel coating material.

Ribose 1,5-bisphosphate regulates rat kidney cortex phosphofructokinase.

Phosphofructokinase (EC 2.7.1.11) is a major enzyme of the glycolytic pathway, catalyzing the conversion of fructose 6-phosphate to fructose 1,6-bisphosphate. In this study, we demonstrated the effect of ribose 1,5-bisphosphate on phosphofructokinase purified from rat kidney cortex. Ribose 1,5-bisphosphate relieved the phosphofructokinase from ATP inhibition and increased the affinity for fructose 6-phosphate at nanomolar concentrations. These activating effects of ribose 1,5-bisphosphate were enhanced in the presence of AMP. Ribose 1,5-bisphosphate reduced the inhibition of the phosphofructokinase induced by citrate. These results suggest that ribose 1,5-bisphosphate is an activator of rat kidney cortex phosphofructokinase and synergistically regulates the enzyme activity with AMP.

Levels of rheumatoid factor isotypes, metalloproteinase-3 and tissue inhibitor of metalloproteinase-1 in synovial fluid from various arthritides.

When synovial effusion is the only symptom, it is often difficult to make an exact diagnosis of the arthritic disease. To distinguish various types of arthritis with synovial effusion, we measured the levels of matrix metalloproteinase-3 (MMP-3, Stromelysin), tissue inhibitor of metalloproteinase-1 (TIMP-1) and rheumatoid factor (RF) isotypes in synovial fluid (SF) from patients with rheumatoid arthritis (RA), osteoarthritis (OA), pyogenic arthritis (PA), pseudogouty arthritis (PG), gouty arthritis (GA) and traumatic arthritis (TA). SF was aspirated from the knee joint or the ankle joint. Levels of IgG-, IgM- and IgA-RF isotypes were measured by ELISA. Levels of MMP-3 and TIMP-1 in SF were simultaneously determined by a one-step EIA system. Levels of IgG-RF, IgM-RF and MMP-3 in SF from RA patients were significantly higher than those in OA, PA, PG, GA and TA. However, IgA-RF in SF from RA patients, when compared with PA and GA, did not show a significantly increased level. In addition, TIMP-1 in SF from RA, when compared with PA and TA, also has not shown a significantly increased level. Therefore, in addition to analysing clinical data, measurements of IgG-RF, IgM-RF and MMP-3 in SF may contribute in distinguishing RA from other arthritic diseases.

Outcomes for patients undergoing one or more total hip and knee arthroplasties.

Either total hip arthroplasty (THA), total knee arthroplasty (TKA) or both were performed in 105 patients from 1981 to 1994. These patients were experiencing severe joint destruction in the lower extremities due to rheumatoid arthritis (RA). These patients were followed for more than 2 years after their last operation. Eighty-six patients were alive and 19 patients had died at the time of follow-up. The 86 living patients were divided into four groups based on the number of replaced joints. Their pre- and postoperative conditions, including such factors as pain, mobility and disability for the quality of life (QOL), were compared. All of the four groups showed some reduction in pain and disability, and an improvement in ambulation after the operations. The 19 deceased patients were classified into two groups, one including those with multiple (three or four) arthroplasties and the other, those with only a small number (one or two). The mean age at death was lower (55.7+/-6.2 years) in patients with multiple arthroplasties than that (69.1+/-7.5 years) in patients with only a small number of arthroplasties. Secondary diseases from RA, such as amyloidosis, spinal injury and pulmonary fibrosis, were found to be the primary cause of death in patients with multiple arthroplasties. The most important finding in this study is that although RA patients with multiple arthroplasties in the lower extremities improved their QOL, they were still afflicted with secondary diseases derived from RA and experienced complications that could shorten their lifespan.

Interleukin-16 in synovial fluids from cases of various types of arthritis.

OBJECTIVES: To examine the characteristic relationship between interleukin-16 (IL-16) and clinical data in various types of arthritis. METHODS: We measured IL-16 levels of the synovial fluids (SF) of patients with various types of arthritis, which included rheumatoid arthritis, traumatic arthritis, pseudogouty arthritis, gouty arthritis, and osteoarthritis, by an enzyme immunosorbent assay, and examined their correlations with clinical parameters. RESULTS AND CONCLUSIONS: Higher levels of IL-16 in synovial fluid from patients with rheumatoid arthritis, traumatic arthritis, and pseudogouty arthritis, compared to those with osteoarthritis, and gouty arthritis were indicated. Also, synovial IL-16 levels in patients with rheumatoid arthritis correlated significantly, especially with synovial matrix metalloproteinase-3 levels. But the IL-16 levels of both synovial fluid and peripheral blood did not correlate with conventional inflammatory parameters such as C-reactive protein, erythrocyte sedimentation rate, or rheumatoid factor. Although the function of IL-16 in inflammatory arthritis has not yet been defined, these data indicated some essential features of IL-16.