Interplay of physical activity and vitamin D receptor gene polymorphism on bone mineral density.

The interplay of physical activity and vitamin D receptor (VDR) gene polymorphism in their effects on bone mineral density (BMD) was studied for 120 Japanese girls aged 18-19 years. BMD at distal radius in the group with the VDR genotype at the Apal site of Aa was significantly higher than that in the aa group, but this association disappeared in a group having the habit of physical activity. The Aa genotype gave a higher BMD than the aa genotype only in the group without the habit of physical activity. The habit of physical activity was associated with a higher BMD only in the aa genotype group. The similar interplay was observed in the VDR genotype at the TaqI site. We thus suggest that physical activity and VDR genotypes affect BMD in independent mechanism to give a saturated level of BMD. Higher dietary calcium intake tended to be associated with higher BMD only in the Aa genotype, suggesting that the calcium intake and VDR genotype affect BMD in a synergistic mechanism.

Novel neuronal nitric oxide synthase (nNOS) selective inhibitor, aplysinopsin-type indole alkaloid, from marine sponge Hyrtios erecta.

Two novel aplysinopsin-type indole alkaloids, 1 and 2, and three known indole alkaloids were isolated from the marine sponge Hyrtios erecta. These compounds exhibited selective inhibitory activity against the neuronal isozyme of nitric oxide synthase (nNOS). Furthermore, new quinolone 7 was also isolated from the same marine sponge. The chemical structures of these new compounds were elucidated on the basis of spectroscopic analysis.

The potent role of increased sympathetic tone in pathogenesis of essential hypertension with neurovascular compression.

OBJECTIVE: To study the role of increased sympathetic tone in pathogenesis of hypertension in patients with essential hypertension with neurovascular compression. METHODS: Twenty-three patients with essential hypertension, 13 patients with secondary hypertension, and 46 normotensive subjects were investigated. Neurovascular compression was evaluated by MRT. The power spectral components of heart rate variability as indices of autonomic nerve tone were determined to investigate the possibility that sympathetic tone mediates the neurovascular compression-induced increase in blood pressure. RESULTS: Neurovascular compression of the rostral ventrolateral medulla (RVLM) was observed in 70% of essential hypertension group, none of secondary hyperension group and 16% of normotensive group (P < 0.001). The age-adjusted low-frequency power spectral density (A-PSD) (0.04 to 0.15 Hz), which is an index of sympathetic tone, was significantly higher in patients with essential hypertension (139.5 +/- 6.7%) with neurovascular compression than in essential hypertension patients without neurovascular compression (92.2 +/- 6.8%), normotensive subjects with (102.8 +/- 13.0%) and without neurovascular compression (100.1 +/- 4.1%), and patients with secondary hypertension (95.7 +/- 10.2%) (P < 0.001). There was no significant difference in the high-frequency A-PSD (0.15 to 0.40 Hz), which is an index of vagal tone, among groups. CONCLUSIONS: Neurovascular compression was not always associated with an increase in sympathetic nerve tone. Hypertension was present in subjects with neurovascular compression, who had increased sympathetic tone but not in those with normal sympathetic tone. An increase in sympathetic tone may mediate the neurovascular compression-induced increase in blood pressure. Journal of Human Hypertension (2000) 14, 807-811

[Chirality of natural products: hyoscyamine and scopolamine].

Monthly changes of the content-ratio between S-(-)- and R-(+)-hyoscyamine as well as those between S-(-)- and R-(+)-scopolamine in the leaves of Datura metel L. cultivated in the field, were quantitatively analyzed by the use of HPLC with a chiral adsorbent. It was found that S-(-)-isomer was predominant for hyoscyamine and the ratio of R-(+)-isomer gradually increased during the growth, whereas in the case of scopolamine, S-(-)-isomer was the sole one found throughout the cultivation period. The 1H-NMR study in the CD3OD solution has suggested that S-(-)-hyoscyamine (1) and S-(-)-scopolamine (2) take a "face-to-face" conformation between their tropane skeletons and the benzene rings of the tropic acid moieties. In the presence of an equimolar NaOD in the CD3OD solution, the racemization at C-2' of 1 and 2 proceeded more rapidly than the hydrolysis at the tropic acid ester bond, presumably due to the steric hindrance caused by their "face-to-face" conformations. In the D2O and H2O solutions, on the other hand, the racemization and the hydrolysis of 1 proceeded smoothly, while those of 2 did not occur. It has been supposed that these individual reaction manners are ascribable in considerable extent to the different basicity of N atom in each tropane skeleton of 1 and 2 and to stronger intramolecular hydrogen bond occurring between the carbonyl oxygen at C-1' and the hydroxyl group at C-3' in the tropic acid moiety of 1.

Noninvasive, continuous evaluation of peripheral vascular resistance in humans.

We sought a noninvasive alternative method of monitoring peripheral vascular resistance continuously in humans, based on the analysis of arterial pressure waveforms. Radial arterial pressure waveforms were recorded noninvasively with a tonometer and analysed using a neural network method. To test the accuracy of this method, the peripheral vascular resistance was also determined by an invasive thermodilution method using a Swan-Ganz catheter in 20 subjects. To test the method in a clinical application, peripheral vascular resistance was determined by the noninvasive method before and after administration of nifedipine in 6 patients with essential hypertension. Neural network analysis of waveforms reliably yielded values between 0.00 and 1.00. Peripheral vascular resistance determined by neural network analysis and according to the invasive method showed a significant (p< 0.005) positive linear correlation. The peripheral vascular resistance measured by neural network analysis showed a significant (p< 0.05) decrease 30 min after administration of nifedipine, paralleling a decrease in blood pressure. Neural network analysis of tonometric radial artery waveforms provides an accurate, noninvasive, and continuous index of peripheral vascular resistance in human subjects. This simple method should permit more extensive homodynamic studies and larger epidemiological surveys in contrast to those undertaken using invasive techniques.

Marine spongean cytotoxins.

Elucidation of naturally occurring toxins is very important from various points of view. Of these toxin studies, search for cytotoxins which show selective toxicity against tumor cells is a challenging subject of study. In search of new bioactive substances from marine organisms, we have been investigating cytotoxic constituents in marine sponges. Through bioassay-guided separation of marine spongean extracts by use of L1210 and KB cell lines, we isolated several potent cytotoxins. This article reviews our recent investigations of three spongean cytotoxins: 1) altohyrtin A and its allied compounds (macrolides, IC50 0.01-0.4 ng/ml (KB cells)) from Hyrtios altum, 2) arenastatin A (a depsipeptide, IC50 5 pg/ml (KB)) from Dysidea arenaria, and 3) callystatin A (a polyketide, IC50 10 pg/ml (KB)) from Callyspongia truncata.

Foods contributing to absolute intake and variance in intake of fat, fatty acids and cholesterol in middle-aged Japanese.

On the basis of 351 one-day weighed diet records, we selected foods/recipes contributing to nutrients of interest for a data-based food frequency questionnaire by contribution analysis and multiple regression analysis. Total fat was largely of animal and vegetable origin, irrespective of analytic methods. Saturated fatty acid was mostly from animal and vegetable sources according to contribution analysis, and that of animal origin was the main contributor by multiple regression analysis. Mono-unsaturated fatty acid was substantially supplied by animal and vegetable products by either analytic method. Poly-unsaturated fatty acid, n-6 poly-unsaturated fatty acid and linoleic acid were found to be of vegetable origin and chicken egg according to contribution analysis; while vegetable oil and mayonnaise were the major contributors to variance in intake. Arachidonic acid was, however, mostly provided by animal sources including chicken egg and fish, irrespective of analytic methods. N-3 poly-unsaturated fatty acids and alpha-linolenic acid were of vegetable and marine origin. Eicosapentaenoic and docosahexaenoic acids were particularly from marine products, irrespective of analytic methods, except for chicken egg in docosahexaenoic acid by contribution analysis. Cholesterol was of animal and marine origin by either analytic method. Thus, foods contributing to absolute intake and variance in intake of fat, fatty acids and cholesterol differed considerably.

Foods contributing to absolute intake and variance in intake of selected vitamins, minerals and dietary fiber in middle-aged Japanese.

Using 351 one-day weighted diet records, we selected foods providing vitamins, minerals and dietary fiber according to contribution analysis (CA) and multiple regression analysis (MRA). Vitamin C was supplied by various vegetables and fruits, and carotene was specifically derived from green-yellow vegetables based on MRA as well as CA. Vitamin A was provided by green-yellow vegetables, fruits, chicken egg and milk (whole) according to CA; whereas chicken liver and pork liver were major sources according to MRA. Vitamin E was mainly of vegetable origin as determined by CA, and largely of spinach, safflower oil and pumpkin as determined by MRA. Vitamin D was mainly derived from chicken egg, fish and mushroom based on CA, and particularly from fish based on MRA. Calcium was supplied by milk (whole), soy products and chicken egg as determined by CA; while milk, tofu and various small fishes were the main contributors to variance. Magnesium was provided by soy products, well-milled rice and spinach according to both analyses, and iron by chicken egg, spinach and soy products. Zinc was largely derived from well-milled rice, followed by chicken egg and milk (whole) as determined by CA, and copper was provided by well-milled rice, soy and wheat products. Dietary fiber was supplied by vegetable sources, whether water soluble or insoluble, based on both analytic methods.

[Variation of chemical constituents in processed licorice roots: quantitative determination of saponin and flavonoid constituents in bark removed and roasted licorice roots].

As a continuing study of chemical characterization of crude drug processing, we have been analyzing the chemical constituents in licorice roots before and after processing. At first, we analyzed chemical constituents in licorice roots of various origins. Next, we have developed the HPLC analytical method, by which saponins and flavonoids, major constituents in various licorice roots, were determined simultaneously in a quantitative manner. In this paper, by means of the HPLC analytical method, chemical constituents in licorice roots, processed and unprocessed, were determined. It was found that nonglycosidic flavonoid constituents were mostly lost while root bark removing, whereas, in roasted licorice roots, sugar chains in the saponin and glycosidic flavonoid constituents were hydrolyzed stepwise during roasting through hydrothermolysis.

Advanced onset of menarche and higher bone mineral density depending on vitamin D receptor gene polymorphism.

Bone mineral density (BMD) at distal forearm, and weight and height of healthy Japanese girls aged 18-19 years were measured and their age at menarche was obtained through a questionnaire. A statistically significant association was found between BMD at distal radius and vitamin D receptor (VDR) gene polymorphism at the ApaI site. The age at menarche in the population with Aa genotype was significantly earlier than that in the aa population. In addition, BMD was significantly dependent on the earlier onset of menarche in the population with genotype Aa but not in the population with genotype aa. BMD was also positively associated with the body mass index (BMI) in the population with genotype Aa. Statistical analysis suggested a stronger effect of VDR genotype on age at menarche than on BMI. Thus, we show that VDR gene polymorphism advances the age at menarche and increases BMD in cooperation with age at menarche.

[Quantitative determination of constituents in various licorice roots by means of high performance liquid chromatography].

During the course of our studies on the chemical constituents of botanically identified Chinese licorice roots of various origins, we have identified 15 kinds of saponins and 49 kinds of flavonoids (including flavonoid glycosides). With these chemical constituents at hand, we have carried out the chemical evaluation studies of licorice roots available in Japan by means of high performance liquid chromatography (HPLC). In the HPLC analysis using an ODS reversed-phase column, a combination of 1% aqueous acetic acid and acetonitrile was used as the mobile phase in a gradient solvent system, and the eluate was monitored with UV absorption at 254 nm (for saponins with a diene or a conjugated carbonyl moiety, liquiritin, and liquiritin apioside) or at 350 nm (for other flavonoid constituents). By our analytical method described here, quantitative analyses of saponin and flavonoid constituents were effected simultaneously. We have found an interesting piece of information concerning the origin of parent plants and their characteristic constituents such as saponins and flavonoids.

Development of data-based semi-quantitative food frequency questionnaire for dietary studies in middle-aged Japanese.

BACKGROUND: We designed a data-based semi-quantitative food frequency questionnaire to clarify the relationship between food intake and lifestyle-related diseases among middle-aged Japanese. METHODS: A total of 351 middle-aged individuals were recruited to a one-day weighed diet record survey in 1994. In all, 586 foods were consumed. Intake of 31 nutrients including energy, protein, fat, carbohydrate, vitamins, minerals and dietary fiber by food was computed by multiplying the weight of food consumed by its nutrient content. First, 252 foods with up to 90 cumulative % contribution to nutrient intake were selected. Of these, foods having apparently the same/similar nutrient content were combined into 206 foods by research dietitians. Next, 183 foods with up to 0.90 cumulative multiple regression coefficient and 90 cumulative % contribution were chosen. At this stage an additional food grouping was made. RESULTS: Finally, 102 foods/recipes were included in the questionnaire: rice (2 items), bread and noodles (11), eggs, milk and dairy products (10), soybean, soybean products and other beans (7), meat including beef, pork and chicken (12), fish (5), other fish, shellfish and fish products (10), green-yellow vegetables (8), other vegetables and mushrooms (7), edible roots (2), seaweeds (3), seeds (2), fruits (8), beverages (7) and confectioneries (8). The frequencies were classified into eight categories. Portion size was calculated for the respective foods largely from the one-day weighed diet record. CONCLUSIONS: The developed semi-quantitative food frequency questionnaire substantially covered the intake of 31 nutrients and may be competent to rank middle-aged Japanese efficiently.

[On the constituents of the root of fukuchiyama-jio, the hybrid of Rehmannia glutinosa var. purpurea and R. glutinosa forma hueichingensis].

In order to characterize the pharmaceutical utility of the root of Fukuchiyama-jio, which was produced as the hybrid of Rehmannia glutinosa LIBOSCH. var. purpurea MAKINO (Akaya-jio in Japanese) and Rehmannia glutinosa LIBOSCH. forma hueichingensis HSIAO (Kaikei-jio in Japanese) (Scrophulariaceae), we have investigated the chemical constituents of the root of Fukuchiyama-jio in detail in comparison with those of Akaya-jio and Kaikei-jio. Chemical analysis as well as quantitative one by means of gas liquid chromatography (GLC) have shown that the root of Fukuchiyama-jio contains a larger amount of iridoid glucosides as compared with Akaya-jio and Kaikei-jio. During these chemical analyses, a new ionone glucoside named oxyrehmaionoside B (7) along with melasmoside (6) were isolated as the characteristic constituents of Fukuchiyama-jio and the structure of 7 has been determined.

Structures of new acylated oleanene-type triterpene oligoglycosides, theasaponins E1 and E2, from the seeds of tea plant, Camellia sinensis (L.) O. Kuntze.

Two new acylated oleanene-type triterpene oligoglycosides, theasaponins E1 and E2, were isolated from the seeds of tea plant [Camellia sinensis (L.) O. Kuntze]. The structures of theasaponins E1 and E2 were elucidated on the basis of chemical and physicochemical evidence to be expressed as 21-O-angeloyl-22-O-acetyltheasapogenol E 3-O-[beta-D-galactopyranosyl(1-->2])[beta-D-xylopyranosyl(1-->2)-alpha-L - arabinopyranosyl (1-->3)]-beta-D-glucopyranosiduronic acid and 21-O-angeloyl-28-O-acetyltheasapogenol E 3-O-[beta-D-galactopyranosyl(1-->2)][beta-D-xylopyranosyl (1-->2)-alpha-L-arabinopyranosyl (1-->3)-beta-D-glucopyranosiduronic acid, respectively. Theasaponin E1 was found to show antisweet activity.

Indonesian medicinal plants. XXI. Inhibitors of Na+/H+ exchanger from the bark of Erythrina variegata and the roots of Maclura cochinchinensis.

Through bioassay-guided separation of the methanol extracts of Indonesian medicinal plants, three inhibitors of the Na+/H+ exchange system, erythrinin B (2), euchrenone b10 (3), and 1,3,5-trihydroxy-4-(3-methylbut-2-enyl)xanthen-9-one (4), were isolated from the bark of Erythrina variegata (Fabaceae) (for 2) and the roots of Maclura cochinchinensis (Moraceae) (for 2, 3, 4). Compounds 2, 3, and 4 significantly inhibited the Na+/H+ exchange system of arterial smooth muscle cells, with minimum inhibitory concentrations of 1.25, 1.25, and 10 micrograms/ml, respectively. Three new prenylated xanthones named isocudraniaxanthones B (5) and A (7) and isoalvaxanthone (9) were also isolated from M. cochinchinensis and the chemical structures were elucidated on the bases of their chemical and physicochemical properties.

Inhibition by ginsenoside Rg3 of bombesin-enhanced peritoneal metastasis of intestinal adenocarcinomas induced by azoxymethane in Wistar rats.

The effects of concomitant use of bombesin and ginsenoside Rg3 on the incidence of peritoneal metastasis of intestinal adenocarcinomas induced by azoxymethane were investigated in male inbred Wistar rats. From the start of the experiment, rats were given weekly s.c. injections of azoxymethane (7.4 mg/kg body weight) for 10 weeks and s.c. injection of bombesin (40 microg/kg body weight) every other day, and from week 20, s.c. injections of ginsenoside Rg3 (2.5 or 5.0 mg/kg body weight) every other day until the end of the experiment in week 45. Bombesin significantly increased the incidence of intestinal tumors and cancer metastasis to the peritoneum in week 45. It also significantly increased the labeling index of intestinal cancers. Although administration of a higher dose of ginsenoside Rg3 with bombesin had little or no effect on the enhancement of intestinal carcinogenesis by bombesin, the location, histologic type, depth of involvement, infiltrating growth pattern, labeling and apoptotic indices and tumor vascularity of intestinal cancers, it significantly decreased the incidence of cancer metastasis. These findings indicate that ginsenoside Rg3 inhibits cancer metastasis through activities that do not affect the growth or vascularity of intestinal cancers.

Indonesian medicinal plants. XIX. 1) Chemical structures of four additional resin-glycosides, mammosides A, B, H1, and H2, from the tuber of Merremia mammosa (Convolvulaceae).

Four new resin-glycosides, named mammosides A (10), B (11), H1 (12), and H2 (13), were isolated from the tuber of Merremia mammosa (Convolvulaceae), a Jamu raw material. Their chemical structures have been elucidated on the bases of chemical and physicochemical evidence, including synthesis of a glycosidic acid designated as mammoside I.

Bruceine B, a potent inhibitor of leukocyte-endothelial cell adhesion.

Leukocyte adhesion to vascular endothelial cells is an essential step in the development of inflammatory diseases. We have searched for inhibitors of leukocyte-endothelial cell adhesion that could be used as anti-inflammatory drugs and found that bruceine B (0.2 microgram/ml; 0.44 microM) inhibited human neutrophil or T cell adhesion to tumor necrosis factor-alpha (TNF) stimulated human umbilical vein endothelial cells (HUVEC). The inhibition of neutrophil adhesion to TNF-stimulated HUVEC by bruceine B was not derived from cytotoxic effects, as determined by measurement of the level of lactate dehydrogenase (LDH) activity in conditioned medium. The effect of bruceine B on neutrophil adhesion to HUVEC was not seen when the neutrophils were preincubated with bruceine B. However, inhibitory effects were evident when the HUVEC were preincubated with bruceine B. Bruceine B also inhibited neutrophil adhesion to lipopolysaccharide-stimulated HUVEC and T cell adhesion to TNF-stimulated HUVEC. These findings suggest that bruceine B may have anti-inflammatory activity.

Metallothionein-independent hepatoprotection by zinc and sakuraso-saponin.

Hepatoprotective activities of zinc and sakuraso-saponin against toxicity of carbon tetrachloride were investigated in metallothionein (MT)-deficient mice. Pretreatment of control 129/Sv mice with zinc or sakuraso-saponin blocked carbon tetrachloride-induced elevation of plasma transaminase activities. Quantitatively equivalent protection against carbon tetrachloride-induced hepatic damage was also observed in MT-deficient mice. Zinc and sakuraso-saponin caused elevation of hepatic MT levels in control 129/Sv mice, whereas hepatic MT was undetectable in MT-deficient mice. To examine the possibility that sakuraso-saponin-induced hepatoprotection is mediated by endogenous zinc, the hepatic concentration of zinc was analyzed. Hepatic zinc concentration in MT-deficient mice was not changed by the treatment of sakuraso-saponin. Injection of sakuraso-saponin caused a decrease of activity of aniline hydroxylation. The suppression of cytochrome P450 appears to be a mechanism by which sakuraso-saponin protects mice from the hepatotoxic effects of carbon tetrachloride. These findings indicate that the hepatoprotective activity of zinc or sakuraso-saponin is not dependent on their MT-inducing activity.