Expression of dentin matrix protein 1 (DMP1) in nonmineralized tissues

Dentin matrix protein 1 (DMP1) is an Arg-Gly-Asp-containing acidic phosphoprotein that was originally identified from a rat incisor cDNA library and was thought to be a dentin-specific protein. DMP1 was later shown to express in a number of hard tissue-forming cells, including osteoblasts, osteocytes, ameloblasts, and cementoblasts, and was considered to play important roles in mineralization. Further, DMP1 gene expression was also detected in fetal bovine brain and in newborn mouse brain. These findings indicate the possibility of DMP1 expression in other soft tissues. In the present study, to clarify the significance of DMP1 expression in nonmineralized tissues, we made a specific antibody to mouse DMP1 peptides and demonstrated that DMP1 protein was localized in mouse brain, pancreas, and kidney by immunohistochemistry. Further DMP1 mRNA was detected in nonmineralized mouse tissues including liver, muscle, brain, pancreas, and kidney by RT-PCR. Based on the evidence that the localization and the expression of DMP1 are not restricted to mineralized tissues, we assume that DMP1 may have functions other than the regulation of mineralization.     

Projections from estrogen receptor-alpha immunoreactive neurons in the periaqueductal gray to the lateral medulla oblongata in the rhesus monkey

The periaqueductal gray (PAG) contains numerous estrogen receptor-alpha immunoreactive (ER-alpha IR) neurons that are distributed in a species-specific way. These neurons might modulate different types of behavior that are mediated by the PAG such as active and passive coping responses, analgesia, and reproductive behavior. In primates, it is not known whether ER-alpha IR PAG neurons represent local interneurons and/or neurons that project to brainstem areas that control these behaviors. In this double labeling study, we asked whether ER-alpha IR neurons in the PAG of the rhesus monkey project to the nucleus retroambiguus (NRA), an area in the ventrolateral caudal medulla oblongata that is involved in expiration, vocalization, and reproductive behavior. Tracer was injected into the caudal lateral medulla oblongata to retrogradely label PAG neurons, and ER-alpha was visualized immunohistochemically. Although ER-alpha IR neurons and NRA-projection neurons were present at similar levels of the PAG, their distributions hardly overlapped. ER-alpha IR PAG neurons that project to the lateral caudal medulla represented less than 2% of ER-alpha IR PAG neurons. These double-labeled neurons were mainly located in the ipsilateral caudal PAG. The cluster of neurons in the medial part of the lateral PAG that projects specifically to the NRA-region did not contain double-labeled cells. The results indicate that only a few ER-alpha IR PAG neurons project to the NRA-region. This might be related to the modest effects of estrogen on mating-related behavior in primates compared most other mammalian species. Remaining ER-alpha IR PAG neurons might act locally on other PAG neurons, or they might represent neurons that project to other areas. Furthermore, the finding that the distributions of ER-alpha IR neurons and neurons that project to premotor neurons in the NRA-region scarcely overlap illustrates that the PAG in primates is very highly organized into anatomically distinct regions compared with other species.     

Reproductive function after treatment of malignant germ cell ovarian tumors

The outcome and reproductive function were examined among patients with malignant ovarian germ cell tumors treated since 1980. Between 1980 and 2001, fertility-sparing surgery was performed in 26 women, 23 of whom received adjuvant chemotherapy. With a median follow-up of 66.6 months, all patients have been alive, with histological types of 6 immature teratomas, 8 dysgerminomas, 6 yolk sac tumors, and 6 mixed types. Clinical stages were involved of 17 early stage and 9 advanced stage patients. After treatment, 20 women out of 26 recovered menstruation within 6 months. During follow-up, two chemotherapy-untreated and one treated patients experienced 4 conceptions in total. A treated patient conceived but selected artificial termination by affection of chemotherapy. Conservative surgery with adjuvant chemotherapy is the standard approach to treat patients with malignant ovarian germ cell tumors. In these 20 years, we experienced no delivery, so that fertility seems to be seriously affected by treatments.     

Angioscopic evaluation of stabilizing effects of an antilipemic agent,bezafibrate, on coronary plaques in patients with coronary artery disease: a multicenter prospective study

To evaluate the stabilizing effects of an antilipemic agent, bezafibrate, on coronary plaques, we carried out a prospective angioscopic and angiographic open trial. From April 1997 to December 1998, 24 patients underwent coronary angioscopy of plaques in non-targeted vessels during coronary interventions and then again 6 months later. The patients were divided into control (10 patients, 14 plaques) and bezafibrate (14 patients, 21 plaques) groups. Oral administration of bezafibrate (400 mg/day) was started immediately after the intervention and was continued for 6 months. The vulnerability score was determined based on the angioscopic characteristics of plaques and compared before and 6 months later. Six months later, the vulnerability score was reduced (from 1.6 to 0.8; P<0.05) in the bezafibrate group and unchanged (from 1.4 to 1.3; NS) in the control group. In the bezafibrate group, the changes in the vulnerability score were not correlated with those in % stenosis or minimal lumen diameter. The plasma total cholesterol level (T-C) was unchanged, triglyceride level (TG) was decreased, and high density lipoprotein cholesterol level (HDL-C) was increased in the bezafibrate group, but were unchanged in the control group. In the bezafibrate group, T-C and TG were decreased and HDL-C was increased in patients with a reduced vulnerability score but were unchanged in those with an unchanged score. These results indicate that 6 month administration of bezafibrate stabilizes coronary plaques and that the stabilization is not correlated with angiographic changes.     

Aging-related changes in release of growth hormone and luteinizing hormone in female rhesus monkeys

A decline in somatic function with aging in women is associated with a decrease in GH release and a loss of estrogen after menopause. As an initial step to establish a monkey model for the neuroendocrine mechanisms underlying somatopause and menopause, we have conducted three experiments in unrestrained aged (approximately 25.7-yr-old) and young (approximately 5.4-yr-old) female rhesus monkeys. GH release was pulsatile, and mean GH release and pulse amplitude were significantly lower in aged monkeys than in young monkeys. Injection of GHRH alone, GH-releasing peptide-2 alone, or the combination of both induced an increase in GH release in both age groups. The mean LH level, pulse amplitude, and baseline LH levels were significantly higher in aged animals than in young animals. Both estrogen and IGF-I levels were lower in aged than young monkeys. These results suggest that in female rhesus monkeys 1) there is a clear decline in circulating GH and IGF-I levels with aging; 2) GHRH and GH-releasing peptide-2 stimulate GH release synergistically; and 3) circulating LH levels increase as estrogen decreases with aging. These results indicate that the rhesus monkey is an excellent model for studies of the neuroendocrine mechanisms of aging.     

Therapeutic usefulness of Keishi-bukuryo-gan for diabetic nephropathy

Keishi-bukuryo-gan is a traditional herbal medicine, which is used clinically as a vascular system disorder-eliminating drug. In this study, its effect on the progression of diabetic nephropathy in experimental rats was investigated. The diabetic nephropathy model used in this study shows functional and morphological changes of the kidney resembling those seen in patients with diabetic nephropathy. Increased proteinuria and serum urea nitrogen and creatinine levels and decreased creatinine clearance, which are important parameters of renal function, were observed in rats with diabetic nephropathy. Pathological examination of the kidney revealed diffuse, nodular and exudative lesions and arteriolar hyalinosis. The deterioration of renal function was ameliorated in rats treated with Keishi-bukuryo-gan for 15 weeks and these results agreed with the renal histological findings. In addition, metabolic abnormalities mediated by persistent hyperglycaemia (the glycation reaction, excessive polyol pathway activity, oxidative stress and lipid metabolic abnormalities) were also observed. However, Keishi-bukuryo-gan reduced accumulation of advanced glycation end products, determined by measuring fluorescence, and serum lipid peroxidation, triglyceride and total cholesterol levels dose-dependently. Thus, this study indicates the potential therapeutic usefulness of Keishi-bukuryo-gan for retarding the progression of renal damage and suggests that its beneficial effects were due to its ability to improve metabolic abnormalities associated with diabetes.     

Clinical evaluation of the effect of daio (rhei rhizoma) on the progression of diabetic nephropathy with overt proteinuria

We studied the effect of traditional herbal medicines containing daio (Rhei Rhizoma) on the long-term progression of diabetic nephropathy with overt proteinuria in eight patients [mean age 60 (45-73) years; duration of diabetes 18 (7-36) years]. At the beginning of the study, mean HbA1c was 8.2% and mean serum creatinine was 1.0 +/- 0.3 mg/dl. Everypatient had diabetic neuropathy and retinopathy. Three of the patients had hypertension and four had ischemic heart disease. After 107 +/- 25 months, the mean serum creatinine level had significantly increased to 4.8 +/- 2.6 mg/dl. The mean serum creatinine levels of five patients not advancing to dialysis treatment increased from 1.2 +/- 0.3 to 3.2 +/- 1.0 mg/dl, and the three patients requiring dialysis increased from 0.8 +/- 0.1 to 7.5 +/- 2.1 mg/dl. In the control group, treated without traditional herbal medicines, the mean serum creatinine level had significantly increased from 1.0 +/- 0.3 to 9.5 +/- 1.9 mg/dl after 71 +/- 12 months. All of the control group required dialysis treatment. Diabetic nephropathy with overt proteinuria is reported to develop into renal failure after 6-7 years. In this retrospective study, traditional herbal medicines with Daio were considered to be effective in prolonging the pre-dialysis period of diabetic nephropathy.     

Inhibitory effects of Oren-Gedoku-To (Huanglian-Jie-Du-Tang) on free radical-induced lysis of human red blood cells

Oren-gedoku-to (Huanglian-Jie-Du-Tang, OGT) has been used for the treatment of cerebrovascular disease, hypertension, gastritis and liver disease in Japan. The present study was to test our hypothesis that ingestion of Oren-gedoku-to extract (TJ-15) would protect red blood cell (RBC) membrane from free radical-induced oxidation if antioxidants in OGT could be absorbed and circulated in blood. When incubated with RBC suspension, OGT and its four constituting herbs provided strong protection for RBC membrane to hemolysis induced by 2,2-azo-bis (2-amidinopropane) dihydrochloride (AAPH), an azo free radical initiator. The inhibitory effect was in a dose-dependent manner at concentrations of 5 microgram/ml to 500 microgram/ml. Furthermore, the ingestion of 7.5 g of OGT (daily dose) was associated with a significant decrease in susceptibility of RBC to hemolysis in humans. The direct protection of RBC membrane from free-radical attack as observed in the present study could provide an important pathophysiological basis for making use of the favorable hemorheological effect of OGT.     

Inhibitory effects of triterpenes isolated from Hoelen on free radical-induced lysis of red blood cells

Hoelen, sclederma of Poria cocos Wolf, has long been used as a sedative and diuretic in traditional medicine. Formerly, we demonstrated that Hoelen in vitro protects red blood cells from AAPH-induced hemolysis. In this study, tests were carried out to identify the main ingredient of Hoelen that has the scavenging effect on free-radicals. Triterpene carboxylic acids isolated from the methanol extract of Hoelen, i.e. pachymic acid, polyporenic acid, 3-epidehydrotumulosic acid, 3beta-hydroxylanosta-7,9(11), 24-trien-21-oic acid and 3-o-acetyl-16 alpha -hydroxytrametenolic acid, were found to have inhibitory activities against AAPH-induced lysis of red blood cells.     

Inhibitory effects of ursodeoxycholic acid on the induction of nitric oxide synthase in vascular smooth muscle cells

The expression of inducible nitric oxide synthase (iNOS) and the resultant increased nitric oxide production are associated with endotoxemia and atherosclerotic lesions observed in transplant hearts or balloon-injured artery. Ursodeoxycholic acid has been shown to have cardiovascular protective effects, such as inhibition of the development of transplant arteriosclerosis, but its mechanism remains unclear. Here, we investigated the effects of ursodeoxycholic acid on nitric oxide production and the expression of iNOS in vascular smooth muscle cells isolated from adult rat aorta and rabbit coronary artery. Nitrite released from cells in the culture medium was measured with the Griess reaction. iNOS mRNA and protein were measured by Northern and Western blot analyses. Treatment with ursodeoxycholic acid (30-1000 microM) significantly inhibited lipopolysaccharide plus interferon-gamma-induced nitric oxide production in a concentration-dependent manner, but ursodeoxycholic acid showed only small inhibitory effects on nitric oxide production that had already been induced by lipopolysaccharide plus interferon-gamma. Ursodeoxycholic acid by itself did not affect basal nitric oxide production. Ursodeoxycholic acid also suppressed lipopolysaccharide plus interferon-gamma-induced expression of iNOS mRNA and protein. Ursodeoxycholic acid had the most potent inhibitory effect among various kinds of bile acids examined, i.e. chenodeoxycholic acid, deoxycholic acid, cholic acid and conjugated bile acids such as tauroursodeoxycholic acid. These results suggest that ursodeoxycholic acid inhibits the induction of iNOS and then nitric oxide production in aortic and coronary artery smooth muscle cells, suggesting a possible mechanism for the cardiovascular protective effect of ursodeoxycholic acid under various pathophysiological conditions such as endotoxemia and atherosclerosis.