EFFECT OF SEASONAL CHANGES ON TESTICULAR MORPHOLOGY AND THE EXPRESSION OF CIRCADIAN CLOCK GENES IN JAPANESE WOOD MICE (APODEMUS SPECIOSUS).

This study aimed to determine the seasonality of reproduction throughout the year in Japanese wood mice (Apodemus speciosus). The effect of seasonal changes on testicular morphology and the periodic expression of circadian clock genes in the hypothalamus and testes of male individuals was evaluated. We also examined the morphology of the testes and caudae epididymides of male mice. In addition, RT-PCR analysis was carried out with mRNA extracted from the hypothalamus and testes to evaluate the expression of the circadian clock genes Clock, Bmal1, Per1, and Cry1. The complete induction of testicular activity was detected from February to April and from August to October, with testes weight increasing with the completion of spermatogenesis (reproductive season). From May to early June and from November to early January, testicular weight declined, the seminiferous tubules reduced in size, spermatogenesis was arrested, and sperm were not produced (non-reproductive season). From mid- June to July and mid-January, the re-induction of testicular activity for spermatogenesis was observed in the seminiferous tubules (transitional season). Out of the four examined genes, Cry1 had the highest expression level in both the hypothalamus and testes throughout the year, followed by Bmal1, Per1, and Clock. The expression of Bmal1 was significantly lower in the hypothalamus and testes during the transitional season compared to the reproductive and non-reproductive seasons. Cry1 transcript levels were also significantly lower in the hypothalamus and testes during the transitional season compared to the reproductive season. In conclusion, the results indicating changes in testicular morphology revealed annual reproductive, non-reproductive, and transmission periods in Japanese wood mice. When an increase in testicular activity was observed indicating the onset of the reproductive season, the mean day length was approximately 11–13 h. The expression of the circadian clock genes Bmal1 and Cry1 in the hypothalamus and testes during the reproductive season was significantly higher than that of the same genes during the transitional season. Consequently, completion of spermatogenesis occurred in the seminiferous tubules of Japanese wood mice testes during the reproductive period.

Antidiabetic activity of the rhizoma of Anemarrhena asphodeloides and active components, mangiferin and its glucoside.

The antidiabetic activity of the rhizoma of Anemarrhena asphodeloides was investigated in KK-Ay mice, an animal model of genetic type 2 diabetes. The water extract of the rhizoma (AA) (90 mg/kg) reduced blood glucose levels from 570 +/- 29 to 401 +/- 59 mg/dl 7 h after oral administration (p<0.05) and also tended to reduce serum insulin levels in KK-Ay mice. AA-treated KK-Ay mice had significantly reduced blood glucose levels in an insulin tolerance test. Based on these results, the antidiabetic mechanism of AA may be due to decreased insulin resistance. In addition, the active components of AA were confirmed to be mangiferin and its glucoside.

Antidiabetic activity of a xanthone compound, mangiferin.

Mangiferin (MF) isolated from Anemarrhena asphodeloides Bunge rhizome, was tested for antidiabetic activity in KK-Ay mice, an animal model of type-2 diabetes. MF lowered the blood glucose level of KK-Ay mice 3 weeks after oral administration (p < 0.01). However, no effect on the blood glucose level in normal mice was seen, indicating that MF could be useful in treating type-2 diabetes. In addition, MF improved hyperinsulinemia and, on insulin tolerance test, reduced blood glucose levels of KK-Ay mice. From these findings, it seems likely that MF exerts its antidiabetic activity by decreasing insulin resistance.

Hypoglycemic activity of the fruit of the Momordica charantia in type 2 diabetic mice.

The antidiabetic activity of Momordica charantia L. (Cucurbitaceae) was investigated in KK-Ay mice, an animal model with type 2 diabetes with hyperinsulinemia. The water extract of the fruit of Momordica charantia L. (MC) reduced the blood glucose of KK-Ay mice 3 weeks after oral administration (p<0.01) and also significantly lowered the serum insulin of KK-Ay mice under similar conditions (p<0.01). However, MC did not affect the blood glucose in normal mice. MC-treated KK-Ay mice blood glucose significantly decreased in an insulin tolerance test. Moreover, the muscle content of facilitative glucose transporter isoform 4 (GLUT4) protein content in the plasma membrane fraction from muscle significantly increased in the orally MC-treated mice when compared with that of the controls (p<0.01). These results suggest that the antidiabetic effect of MC is derived, at least in part, from a decrease in insulin resistance because of the increase of GLUT4 protein content in the plasma membrane of the muscle.

Infarct size limitation by bradykinin receptor activation is mediated by the mitochondrial but not by the sarcolemmal K(ATP) channel.

Earlier studies have shown that activation of bradykinin B2 receptor triggers protein kinase C (PKC)-mediated cardioprotective mechanism in ischemic preconditioning (PC). In the present study, we examined whether the effector in this B2-receptor triggered pathway of PC is the ATP sensitive potassium (K(ATP)) channel in the mitochondria (mito-K(ATP) channel) or K(ATP) channel in the sarcolemma (sarc-K(ATP) channel). Isolated rabbit hearts were perfused with modified Krebs-Henseleit buffer in a Langendorff mode, and regional myocardial ischemia was induced by occluding a left coronary artery for 30 min and then reperfusing for 2 hours. Infarct size was determined by triphenyltetrazolium chloride staining and expressed as a percentage of area at risk (% IS/AR). Infusion of bradykinin (500 nmol/L) for 15 min prior to ischemia significantly reduced % IS/AR from 37.4 +/- 2.9 (SE) of the untreated controls to 12.0 +/- 3.3%. This protective effect of bradykinin was completely abolished by coinfusion of 5-hydroxydecanoate (5-HD, 50 micromol/L), a selective mito-K(ATP) channel blocker (% IS/AR = 44.2 +/- 6.4). In contrast, a high dose of HMR1098 (20 micromol/L), which is a newly developed sarc-K(ATP) channel selective blocker with IC50 of 0.6 micromol/L, failed to modify the infarct size limitation by preischemic infusion of bradykinin (% IS/AR = 11.7 +/- 3.4). Neither 5-HD nor HMR1098 alone modified infarct size (% IS/AR = 37.8 +/- 3.8 and 35.1 +/- 6.2, respectively). These results suggest that opening of the mito-K(ATP) channel but not the sarc-K(ATP) channel is involved in infarct size limitation by a mechanism triggered by bradykinin B2 receptor activation.

[Intra-arterial infusion chemotherapy in combination with microwave hyperthermia for cancer of head of pancreas and liver metastasis--a case of 16 years survival].

Our experience of arterial infusion chemotherapy combined with regional hyperthermia in the treatment of non-resectable pancreatic cancer was presented. A patient with cancer in the pancreatic head with accompanying extensive metastasis in both hepatic lobes was treated by sub selective aortic infusion of 5-FU and MMC with microwave hyperthermia. Both the cancer in the pancreatic head and the liver metastasis showed complete remission for 16 years.

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.

Antioxidant activity of soya hypocotyl tea in humans.

Antioxidative activity of isoflavones has not been shown in humans. Newly-developed isoflavone-rich soya hypocotyl tea contains about 12 mg isoflavones per liter. 15 tea drinkers and 23 control young female students were randomly selected from volunteers, and underwent physical examination, blood chemistry and urinary analysis before and after one month of tea drinking. A three-day dietary record was taken before each physical examination. The tea drinkers showed a lower level of phosphatidylcholine hydroperoxide (PCOOH) and phosphatidyl-ethanolamine hydroperoxide (PEOOH) in the red blood cells and a significant reduction of 8-hydroxydeoxyguanine (8ohdG) in the urine compared to the controls.

A comparative study of Embelia schiperi and Embelia keniensis on blood glucose and triglycerides in normal and epinephrine-induced hyperglycemic mice.

Intraperitoneal administration of the methanol extract of Embelia schiperi (ES) to normal mice caused a significant decrease in blood glucose (p < 0.01) and a significant increase in triglycerides 4 hours after administration at 100 mg/kg (p < 0.01). The toluene fraction of Embelia keniensis methanol extract (TS) showed hypoglycemic and lipid lowering activity 7 hours after intraperitoneal administration at 100 mg/kg. In addition, TS (100 mg/kg) administration significantly decreased blood glucose in epinephrine-induced hyperglycemic mice (p < 0.01). Moreover, ES tended to increase while TS tended to decrease the blood triglycerides in epinephrine-induced hyperglycemic mice. On the other hand, no changes in blood cholesterol were observed after the administration of ES or TS in normal and epinephrine-induced hyperglycemic mice. We found that two species from Embelia, ES and TS, have different activities on blood glucose and triglycerides in normal and epinephrine-induced hyperglycemic mice.

Antidiabetic mechanism of Bakumondo-inshi.

To determine the antidiabetic mechanism of Bakumondo-inshi (BI), we examined its effects on glucose absorption, alpha-glucosidase activity, sodium-dependent glucose transporter and facilitative glucose transporter isoform 5 (GLUT5) in small intestine. The oral administration of BI into KK-Ay mice caused a significant decrease in the glucose absorption in small intestine. The small intestine content of active glucose transporter isoform (SGLUT) protein content from KK-Ay mouse significantly decreased in the BI-treated KK-Ay mice compared to that in the controls. However, the small intestine content of facilitative glucose transporter isoform, GLUT5 protein content did not change. The alpha-glucosidase activity in small intestine significantly decreased in the BI-treated KK-Ay mice. These results suggest that the antidiabetic effect of BI is derived, at least in part, from a decrease of glucose absorption in small intestine , due to the reduction of SGLUT protein content in total membrane of the small intestine and the reduction of alpha-glucosidase activity. Because of its therapeutic mechanism, BI could be a new category of therapeutic agent for non-insulin dependent diabetic mellitus.

Effects of Sho-saiko-to, a Japanese herbal medicine, on hepatic fibrosis in rats.

It has been shown that lipid peroxidation is associated with hepatic fibrosis and stellate cell activation. Sho-saiko-to (TJ-9) is an herbal medicine, which is commonly used to treat chronic hepatitis in Japan, although the mechanism by which TJ-9 protects against hepatic fibrosis is not known. As a result, we assayed the preventive and therapeutic effects of TJ-9 on experimental hepatic fibrosis, induced in rats by dimethylnitrosamine (DMN) or pig serum (PS), and on rat stellate cells and hepatocytes in primary culture, and assessed the antioxidative activities and the active components of TJ-9. Male Wistar rats were given a single intraperitoneal injection of 40 mg/kg DMN or 0.5 mL PS twice weekly for 10 weeks. In each model, rats were fed a basal diet throughout, or the same diet, which also contained 1.5% TJ-9, for 2 weeks before treatment or for the last 2 weeks of treatment. TJ-9 suppressed the induction of hepatic fibrosis, increased hepatic retinoids, and reduced the hepatic levels of collagen and malondialdehyde (MDA), a production of lipid peroxidation. Immunohistochemical examination showed that TJ-9 reduced the deposition of type I collagen and the number of alpha-smooth muscle actin (alpha-SMA) positive-stellate cells in the liver and inhibited, not only lipid peroxidation in cultured rat hepatocytes that were undergoing oxidative stress, but also the production of type I collagen, alpha-SMA expression, cell proliferation, and oxidative burst in cultured rat stellate cells. In addition, TJ-9 inhibited Fe2+/adenosine 5'-diphosphate-induced lipid peroxidation in rat liver mitochondria in a dose-dependent manner and showed radical scavenging activity. Among the active components of TJ-9, baicalin and baicalein were found to be mainly responsible for the antioxidative activity. These findings suggest that Sho-saiko-to (TJ-9) functions as a potent antifibrosuppressant by inhibition of lipid peroxidation in hepatocytes and stellate cells in vivo.

Creation of genetic information by DNA polymerase of the thermophilic bacterium Thermus thermophilus.

Genetic information encoded in a template of a genome is replicated in a complementary way by DNA polymerase or RNA polymerase with high fidelity; no creation of information occurs in this reaction unless an error occurs. We report here that DNA polymerase of the thermophilic bacterium Thermus thermophilus can synthesize up to 200 kb linear double-stranded DNA in vitro in the complete absence of added primer and template DNAs, indicating that genetic information is actively created by protein. This ab initio DNA synthesis occurs at 74 degrees C and requires magnesium ion. There is a lag time of approximately 1 h and then the reaction proceeds linearly. The synthesized DNAs have a variety of sequences; they are mostly tandem repetitive sequences, e.g. (CATGTATA) n , (TGTATGTATACATACATA) n and (TATACGTA) n . Some degenerate sequences of these basic repeat units are also found. The similar repetitive sequences are found in many natural genes. These results, together with similar results found using DNA polymerase of archaeon Thermococcus litoralis , suggest that creative, non-replicative synthesis of DNA by protein was a driving force for diversification of genetic information at a certain stage of the evolution of life on the early earth.

Calcium bioavailability of a total bone extract (TBE) and its effects on bone metabolism in rats.

The effects of a total bone extract (TBE), a new mineral preparation made from bovine bone rendered soluble by lactic acid and citric acid under decompression, on the bone metabolism and apparent calcium absorption were examined for an ovariectomized (OVX) rat model of osteoporosis. The apparent calcium absorption from TBE was significantly higher than that from calcium carbonate. There were no significant differences in serum biochemical indices. Although there were no significant differences in the dry weight, ash and calcium content in the tibia, or in the bone mineral density of the femur, TBE feeding increased the cortical thickness index of the femur. A positive effect of TBE on bone formation is thus suggested.

[Transcatheter oily chemoembolization and intermittent hepatic artery infusion chemotherapy in the management of advanced hepatocellular carcinoma].

Between 1990 and 1997, 227 patients with hepatocellular carcinoma were treated by intrahepatic arterial injection of a Lipiodol-Epirubicin-Mitomycin C emulsion followed by intermittent hepatic artery infusion of Epirubicin, Mitomycin C and 5-FU, employing an implantable subcutaneous infusion port. A catheter was inserted percutaneously into the hepatic artery using the Seldinger technique. Objective remission was induced in 80% of the evaluable patients as evidenced by a decrease in their AFP and PIVKA II levels. These remissions were also confirmed by liver sonogram and CT scan showing decreased tumor volume. Transcatheter oily chemoembolization combined with intermittent hepatic artery infusion chemotherapy seems to be an effective treatment for unresectable hepatocellular carcinoma both for palliation of symptoms as well as prolongation of survival with good quality of life.

Requirement of the glycosyl parts in platycodin D to stimulate pancreatic exocrine secretion.

The whole structure of platycodin D is found to be essential to stimulate the volumetric increase in the pancreatic exocrine secretion, while the prosapogenins prepared from platycodin D increased only protein output of pancreatic juice.

Antidiabetic effect of seishin-kanro-to in KK-Ay mice.

The hypoglycemic effect of Seishin-kanro-to (SK) was investigated in KK-Ay mice, one of the non-insulin dependent diabetic mellitus types. SK (1700 mg/kg) reduced the blood glucose of KK-Ay mice from 557 +/- 17 to 383 +/- 36 mg/100 ml 7 hours after single oral administration (P < 0.001). SK also decreased the blood glucose and improved glucose tolerance 5 weeks after repeated administration in KK-Ay mice. These results support, therefore, the use of SK in patients with diabetes and confirm its role as a traditional medicine. In addition, the active plants of SK were identified as the rhizome of Anemarrhena asphodeloides Bunge and the radix of Rehmannia glutinosa Liboschitz.

Hypoglycemic activity of some triterpenoid glycosides.

Four triterpenoid glycosides isolated from the rhizomes of Polygala senega var. latifolia, senegins II-IV (1-3) and desmethoxysenegin II (4), were tested for hypoglycemic activity in normal and KK-Ay mice. Compounds 1 and 2 reduced the blood glucose of normal mice 4 h after intraperitoneal administration and also significantly lowered the glucose level of KK-Ay mice under similar conditions. Compounds 3 and 4, as well as senegose A (5), an oligosaccharide ester, were inactive when tested against normal mice.

Hypoglycemic activity of to-kai-san (Chinese medicines) in normal and KK-Ay mice.

The hypoglycemic effect of To-Kai-San (TS) was studied in normal mice, streptozotocin-induced diabetic mice, one of the animal models of insulin-dependent diabetes mellitus (IDDM) with hypoinsulinemia, and KK-Ay mice, one of the animal models of non-insulin-dependent diabetes mellitus (NIDDM) with hyperinsulinemia. TS (1,500 mg/kg) reduced the blood glucose of the KK-Ay mice from 441 +/- 36 to 285 +/- 52 mg/100 ml 7 h after a single oral administration (p < 0.001). However, no changes in blood glucose were observed in the normal and STZ-induced diabetic mice. TS also decreased the blood glucose 20 days after repeated administration to normal and KK-Ay mice. In addition, TS-treated KK-Ay mice showed decreased plasma insulin levels after both single and repeated administration. From these findings, it is believed that TS may require the presence of insulin to display its activity.

Hypoglycemic effect of the rhizomes of Smilax glabra in normal and diabetic mice.

The hypoglycemic effect of the rhizomes of Smilax glabra ROXBURGH (Liliaceae) was investigated in normal and KK-Ay mice, one of the animal models of non-insulin dependent diabetes mellitus (NIDDM) with hyperinsulinemia. The methanol extract of rhizomes of Smilax glabra ROXBURGH (SM, 100 mg/kg body weight) reduced the blood glucose of normal mice 4 h after intraperitoneal administration (p<0.05), and also significantly lowered the blood glucose of KK-Ay mice under similar conditions (p<0.001). However, SM did not affect the blood glucose in streptozotocin-induced diabetic mice, one of the animal models of insulin-dependent diabetes mellitus (IDDM) with hypoinsulinemia. SM also suppressed epinephrine-induced hyperglycemia in mice. SM-treated KK-Ay mice significantly decreased the blood glucose in an insulin tolerance test. We concluded that the hypoglycemic effect of SM raised insulin sensitivity.

Hypoglycemic action of embelia madagascariensis in normal and diabetic mice.

The hypoglycemic effect of Embelia madagascariensis (Myrsinaceae) was investigated in both normal and streptozotocin-induced diabetic mice. The methanol extract of leaves of Embelia madagascariensis (EL)(500 mg/kg) reduced the blood glucose of normal mice from 206 +/- 9 to 137 +/- 10 mg/100 ml 4 hours after intraperitoneal administration (P < 0.001), and also significantly lowered the blood glucose of streptozotocin-induced diabetic mice from 570 +/- 29 to 401 +/- 59 mg/100 ml under similar conditions (P < 0.05). EL also suppressed epinephrine-induced hyperglycemia in mice (control vs EL, P < 0.01).