The effects of exercise load during development on oxidative stress levels and antioxidant potential in adulthood.

The objective of this study was to elucidate the impact of physical activity during the growth period as well as on oxidative stress and antioxidative potential in adulthood. The experimental animals used were four-week old male Wistar rats, which were randomly divided into three groups. The exercise loads were as follows: control (CON), treadmill exercise (TE), and jumping exercise (JE). The exercise was performed at the same time of day, at a frequency of five days per week, for eight weeks. Derivatives of reactive oxygen metabolites (d-ROSs) and biological antioxidant potential (BAP) were measured during periods of rest prior to commencement of the experiment and after the experiment. Analysis was conducted using a Wilcoxon signed-rank test and Schaffer's multiple comparison procedure and the significance level was set at p < 0.05. The percent increase in d-ROM levels in the JE group, which experienced short-duration intense exercise loads, was higher than that in the TE group, which experienced moderately intense exercise loads. However, BAP, which is an index of antioxidant potential, markedly decreased in adulthood in the CON group, as compared to that in the developmental period, whereas the exercise groups showed no notable changes in BAP levels. Oxidative stress levels and antioxidant potential are affected differently in adulthood, depending on the intensity of sustained exercise loads experienced during development. Results suggested that in order to increase antioxidant potential, while taking oxidative stress production into account, moderately intense exercise loads are more desirable than highly intense exercise loads.

Functional dissociation between Kana and Kanji: agraphia following a thalamic hemorrhage.

We report the case of a 61-year-old woman with a left thalamic hemorrhage causing agraphia of Kanji (morphograms). Single-photon emission computed tomography (SPECT) showed a decrease in the blood flow in the left thalamus from the superior temporal convolution to the parietal lobe, as well as in the frontal lobe while computed tomography showed no remarkable lesions in the cortex. The agraphia in this case may be due to the thalamic lesion itself, but the SPECT findings strongly suggest that a secondary cortical lesion may be involved in producing the higher cognitive disorder.

Thermochemical transformation of sulfur compounds in Japanese domestic Allium, Allium victorialis L.

Sulfur compounds contributed to the health promotion in Allium species are produced via enzymic and thermal reactions. Potent antithrombotic agents which have been identified as allyl trisulfides, dithiins, and ajoene in garlic (A. sativum) and caucas (A. victorialis) are thermochemically transformed from allicin (allyl 2-propenethiosulfinate). The leaves and stems of Japanese domestic Allium plant, A. victorialis L. which is widely distributed in the northern part of Japan, under the name "Gyoja-ninniku" is a nutritious vegetable. The significant flavor compounds of caucas are methyl allyl disulfide (Chinese chive odor), diallyl disulfide (garlic-like odor), and dimethyl disulfide and methyl allyl trisulfide (pickles-like odor) among more than 85 peaks on the gas chromatogram. 2-Vinyl-4H-1,3-dithiin and 3,4-dihydro-3-vinyl-1,2-dithiin as platelet aggregation inhibitors were found eliminated in dichloromethane extract of caucas. The significant health promoting factors, allyl trisulfides and dithiins were relatively increased when caucas was cooked on a frying pan.

Thymidylate synthase levels as a therapeutic and prognostic predictor in breast cancer.

5-Fluorouracil (5-FU) is a commonly used adjuvant therapeutic drug in treating breast cancer. 5-FU is metabolically converted to 5-fluorouracil-2'-deoxyuridine-5'-monophosphate-(FdUMP) which is believed to inhibit DNA synthesis in neoplastic cells by forming a tightly bound ternary complex with thymidylate synthase (TS). In the present study, we examined the possible relationship between TS levels and clinico-pathologic and prognostic features in breast disease. Mean TS levels of 2.9 pmol/g, 6.1 pmol/g, and 23.1 pmol/g were obtained in cases of benign breast disease (3 cases), primary breast cancer (115 cases), and recurrent tumors (4 cases), respectively. In breast cancer, mean TS levels significantly correlated with S-phase fraction (SPF), DNA polymerase a and lymphatic invasion. Thus, TS levels in breast cancer significantly reflected cell proliferation and malignancy. Regarding the survival rate, patients with TS values above 10 pmol/g showed an unfavorable prognosis. The effectiveness of adjuvant 5-FU derivatives chemotherapy was reflected in a higher disease-free survival rate in node (+) cases showing TS levels between 5 and 10 pmol/g (p < 0.1), but not in node (-) cases. In conclusion, TS levels in neoplastic tissues of the breast were highest in recurrent tumors, followed by those in primary cancer, benign breast disease and in breast cancer which reflected proliferative activity. Breast cancers with extremely high TS levels were accompanied by an unfavorable prognosis; however, those with moderately high TS levels tended to respond to adjuvant chemotherapy with 5-FU derivatives.

Expression of carbohydrate-binding protein p33/41 in human tumor cell lines.

We previously reported a new type of lectin, p33/41 (annexin IV), which was isolated from a bovine tissue extract [Kojima, K. et al. (1992) J. Biol. Chem. 267, 20536-20539]. When the expression of p33/41 (annexin IV) was surveyed in the lysates of 39 human tumor cell lines by SDS-PAGE, followed by Western blot analysis with polyclonal anti-bovine p33/41 and monoclonal anti-annexin IV (Z016, Zymed) antibodies, 21 cell lines were found to be reactive with the polyclonal antibody, whereas all 39 cell lines were stained with Z016. These results together with those obtained with standard proteins, annexins IV and V, suggested that the monoclonal antibody, Z016, recognizes annexin V, but not p33/41 (annexin IV). Therefore, we performed cDNA cloning of human p33/41 (annexin IV) to prepare a recombinant protein and raised monoclonal antibodies against the protein. Northern blot analysis with the cDNA as a probe showed that a human colon cancer cell line, HT29, contains p33/41 (annexin IV) mRNA of two sizes, 2.0 and 3.0 kb. The two monoclonal antibodies, AS11 and AS17, against the recombinant protein generated were useful for flow cytometric analysis, ELISA, Western blot analysis and immunoprecipitation. Flow cytometric analysis with AS17 showed that p33/41 (annexin IV) is located in the cytoplasm of HT29 cells, but not on the cell surface. However, one of the cell surface proteins first labeled with biotin and then solubilized with a detergent was immunoprecipitated with AS17. The results suggest the existence of a membrane spanning form of p33/41 (annexin IV).

Effects of spinach leaf protein concentrate on the serum cholesterol and amino acid concentrations in rats fed a cholesterol-free diet.

The effects of spinach leaf protein concentrate (SPPC) on serum and liver lipid concentrations and on serum free amino acid concentrations were examined in rats fed a cholesterol-free diet containing 2 and 10% fats. The serum total cholesterol, triacylglycerol and phospholipid concentrations in the rats fed an SPPC diet containing 2% corn oil were significantly lower than those of the rats fed a corresponding casein diet. When 10% corn oil or lard was used, the serum cholesterol-lowering effect of the SPPC became insignificant, but the serum and liver triacylglycerol concentrations were kept at significantly lower levels. Both the amounts of fecal neutral steroids and bile acids were significantly higher in the rats fed the SPPC than those of the casein-fed rats. The concentrations of serum threonine, serine, glutamine, glycine, cystine, and isoleucine were significantly higher in the rats fed the SPPC diet containing 2% corn oil compared with those of the control rats, but when the dietary fat was raised to 10%, only glycine showed a higher serum concentration. These results indicate that the SPPC has a stronger cholesterol-lowering effect at a lower dietary fat level, 2%, and the activity is partly due to the inhibition of intestinal absorption of cholesterol and bile acid, and partly due to an increase in the concentration of some of the serum amino acids.

Supplemental dietary cystine elevates kidney metallothionein in rats by a mechanism involving altered zinc metabolism.

Dietary sulfur-containing amino acids influence zinc and copper status. This study was conducted to investigate the relationship between the status of these elements and tissue metallothionein in rats fed diets supplemented with sulfur-containing amino acids. In a series of experiments, a diet containing 100 g casein/kg diet was unsupplemented or supplemented with L-cystine (3-50 g/kg diet) or L-methionine (3 or 30 g/kg diet). Kidney concentrations of zinc and copper in rats fed the diet supplemented with high levels of cystine (25 or 50 g/kg) were significantly higher than those in rats fed the unsupplemented diet. Kidney concentrations of metallothionein and metallothionein mRNA were also significantly higher in rats fed cystine-supplemented diets. There was a correlation (r = 0.838, P < 0.01) between the levels of zinc and metallothionein in kidney of rats fed the diets with different levels of cystine and zinc. However, in the rats fed the diets with different levels of cystine and copper, the correlation was not as strong (r = 0.587, P < 0.01). The changes in kidney metallothionein concentration due to the addition of 3 or 30 g/kg cystine were associated with parallel changes in serum zinc concentration and in apparent absorption of zinc, but not in serum copper concentration or apparent absorption of copper. Addition of 3 or 30 g methionine/kg diet to the diet did not affect kidney concentrations of metallothionein or zinc. These results indicate that supplementation of cystine to the diet can induce kidney metallothionein through a mechanism involving altered zinc metabolism.

A simple maneuver in follow-up digital subtraction angiography for multiple coronary-aorta bypass grafts.

A simple maneuver in intraarterial digital subtraction angiography (DSA) is proposed to improve the images of left internal thoracic artery bypass grafts (LITAGs). A contrast flush in the left subclavian artery, using a catheter that had been percutaneously introduced from the left brachial artery, was carried out in 14 patients with multiple coronary-aorta bypass grafts (CABGs) after aortic DSA. The side-hole portion of the catheter tip was positioned in the proximal left subclavian artery as the coiled portion remained in the aortic arch. Nonionic contrast medium (350 mg/ml of iodine) was injected with an automatic injector in a volume of 6-8 ml and at a flow rate of 3-4 ml/sec. Left subclavian arterial flush DSA delineated the LITAGs excellently in 11, well in 3, and fairly or poorly in none of the 14 cases. There were no complications in any of these cases. A supplemental contrast flush of the left subclavian artery with a pigtail catheter following aortic DSA improves the diagnosis of multiple CABG patency.

Role of nitric oxide in the pancreatic blood flow response to caerulein.

To clarify the role of nitric oxide (NO) in the pancreas, blood flow in the rat pancreas (pancreatic blood flow: PBF) was investigated by the hydrogen clearance technique using a specific NO synthase inhibitor, N omega-nitro-L-arginine (L-NNA). Continuous infusion of caerulein at doses of 5 and 20 micrograms/kg/h caused a significant increase in PBF in the early phase of caerulein infusion. The caerulein-induced increase in PBF was not affected significantly by atropine sulfate (100 micrograms/kg), nor by phenoxybenzamine (5 mg/kg) plus propranolol (50 micrograms/kg). Administration of L-NNA (0.5, 5, or 30 mg/kg) did not affect the basal PBF, but at 5 mg/kg it inhibited completely the caerulein-induced increase in PBF. The inhibitory action of L-NNA was reversed by a large dose of L-arginine (100 mg/kg bolus, i.v., followed by a continuous infusion at 400 mg/kg/h), but not by its enantiomer D-arginine. These results strongly suggest that NO has a mediator role in the early phase vascular response of the pancreas to superphysiologic doses of caerulein.

A novel neutral polysaccharide having activity on the reticuloendothelial system from the root of Glycyrrhiza uralensis.

A neutral polysaccharide, named glycyrrhizan UC, was isolated from the root of Glycyrrhiza uralensis Fischer. It was homogeneous on electrophoresis and gel chromatography, and its molecular mass was estimated to be 69,000. Glycyrrhizan UC is composed of L-arabinose:D-galactose:D-glucose:L-rhamnose in the molar ratio of 10:30 27:1. Methylation analysis, carbon-13 nuclear magnetic resonance and periodate oxidation studies indicated its structural feature as an arabino-3,6-galacto-glucan type polysaccharide.

Platelet aggregation caused by Carybdea rastonii toxins (CrTX-I, II and III) obtained from a jellyfish, Carybdea rastonii.

The pharmacological mechanisms of platelet aggregation induced by highly toxic proteins (CrTX-I, CrTX-II, and CrTX-III) obtained from tentacles of a jellyfish, Carybdea rastonii, were investigated. When the partially purified toxin (pCrTX) and CrTXs were added to the citrated platelet-rich plasma (PRP), aggregation was produced in a concentration-dependent manner. The activity of CrTXs was approximately 100 times more potent than pCrTX. The CrTXs-induced aggregation was little affected by indomethacin and quinacrine at concentrations sufficient to inhibit arachidonic acid- and collagen-induced aggregation. The CrTXs-induced aggregation in washed platelets was significantly augmented in the presence of Ca2+. The pretreatment with verapamil failed to modify this augmentation of aggregation. The concentration of cytoplasmic-free calcium ([Ca2+]i) of platelets was increased by CrTXs at the same concentrations that produced aggregation. This effect of CrTXs was again little affected by verapamil. CrTXs at the same concentrations as those that produced aggregation and increased [Ca2+]i caused depolarization of platelets, which was unchanged after pretreatment with sodium or potassium transport inhibitors. CrTX-I significantly increased the 22Na flux into platelets and this effect of CrTX-I was unaffected by tetrodotoxin. The CrTX-I-induced aggregation, depolarization, and increase in [Ca2+]i were all significantly attenuated in the low Na+ medium. These results suggest that CrTXs cause a massive depolarization by increasing cation permeability and this generalized depolarization permits an inward movement of Ca2+ down its electrochemical gradient which, in turn, triggers platelet aggregation.

Delayed flare-up reactions caused by jellyfish.

Four patients had a recurrence of cutaneous lesions 1 week after being stung by jellyfish. Three patients had flare-up lesions after only one exposure to jellyfish. All of the recurring lesions were vesicular erythema, and the histological findings of case 3 corresponded to that of allergic contact dermatitis.

Platelet aggregation caused by a partially purified jellyfish toxin from Carybdea rastonii.

A partially purified toxin (pCrTX) was obtained from the tentacles of the jellyfish, Carybdea rastonii. When pCrTX (3 X 10(-8) - 3 X 10(-7) g/ml) was added to citrated platelet-rich plasma, aggregation was produced in a concentration-dependent manner. Scanning electron microscopic examination revealed that both pCrTX and collagen produced aggregates of platelets possessing many pseudopods. The concentration which produced 50% aggregation for pCrTX was 1.8 X 10(-7) g/ml, as compared to 2.3 X 10(-6) g/ml for collagen. The pCrTX-induced aggregation was only slightly inhibited by indomethacin and quinacrine in concentrations sufficient to inhibit arachidonic acid- and collagen-induced aggregation. pCrTX was less active in washed platelets suspended in Ca2+ free medium, whereas the pCrTX-induced aggregation was significantly augmented in the presence of Ca2+. The augmentation of aggregation by Ca2+ was only slightly attenuated by pretreatment with 100 microM verapamil. pCrTX significantly increased the concentration of cytoplasmic free Ca2+ ([Ca2+]i) and depolarized the platelet membrane in concentrations that produced aggregation. The increase in [Ca2+]i caused by pCrTX was little affected by verapamil. The depolarization by pCrTX was unchanged in the presence or absence of Ca2+, or by sodium or potassium transport inhibitors. The movement of 22Na+ into platelets was significantly increased by pCrTX. This increase in the movement of 22N+ into platelets was unaffected by tetrodotoxin. On the other hand, pCrTX-induced aggregation, depolarization and the increase in [Ca2+]i were all significantly attenuated in low Na+ medium. These results suggest that pCrTX causes a massive depolarization by increasing cation permeability indiscriminately and this generalized depolarization permits an inward movement of calcium down an electrochemical gradient which, in turn triggers platelet aggregation.

Effects of antidepressants in the rat forced swimming test.

Effects of antidepressants and other drugs on the behaviour of rats in the forced swimming test were examined. Acute and chronic administration of antidepressants reduced the duration of immobility during the first 5 or 10 min of a 30 min test by prolonging escape-directed behaviour which appeared only during this period. However, the drugs did not affect the duration of immobility during the last 20 or 25 min, when the rats were in a state of almost complete immobility. In contrast, methamphetamine, caffeine and scopolamine reduced the duration of immobility not only during the first 5 min but also the next 15 or 25 min without prolonging the escape-directed behaviour but by increasing general motor activity. The effect of antidepressants was potentiated by chronic treatment as compared to acute administration. In the chronic experiments, a significant reduction in the duration of immobility was first observed on the 6th day of the treatment. Although a single injection of diphenhydramine caused an effect similar to antidepressants, this effect disappeared after chronic treatment. These results indicate that immobility itself is not affected by antidepressants. However, it is suggested that the reduction in the duration of immobility only during the first 5 min of the test, which was caused by the prolongation of the escape-directed behaviour, and the potentiation of the effect after chronic treatment are an action specific to antidepressants.