CPNE7 Induces Biological Dentin Sealing in a Dentin Hypersensitivity Model.

Dentin hypersensitivity commonly occurs due to opened dentinal tubules for many reasons. In our previous study, copine 7 (CPNE7) could induce dentin formation for an indirect pulp-capping model in vivo. This study aims to investigate the formation of tertiary dentin when CPNE7 is applied to intentionally exposed dentin with nothing over it in vivo, whether it affects microleakage of the teeth, and the penetration ability of CPNE7 molecules through dentinal tubules in vitro. Cervical dentin areas of 6 maxillary incisors of 5 beagles were exposed to a class V-like lesion, and 1 side of 3 maxillary incisors was adapted with recombinant CPNE7 protein for 5 min as the experimental group. The other side was the control group, and there was no treatment of ethylenediaminetetraacetic acid (EDTA) and CPNE7 after preparation. The defects were exposed without any restorations, and all beagles were sacrificed after 4 wk. The fluid penetration of exposed dentin areas was investigated by a microleakage-testing device and confocal laser scanning microscope. Tertiary dentin formation was confirmed with histological scanning electronic microscopic analysis. Tertiary dentin formation reduces dentinal fluid flow due to occluded tubules or discontinuity with primary or secondary dentin. The in vivo hypersensitivity model with the anterior teeth of beagle dogs showed newly formed tertiary dentin at the dentin-pulp boundary in recombinant CPNE7-treated teeth when compared with the untreated control group in histologic analysis. Scanning electronic microscopic analysis revealed occluded sites with mineral deposition of intratubular dentin. In the permeability test, the mean microleakage value of the CPNE7-treated group was significantly lower than that of the control group (P < 0.05). The tubular penetration of rhodamine B-combined CPNE7 was confirmed under confocal laser scanning microscope. CPNE7 induces formation of tertiary dentin through shallowly exposed dentinal tubules, which decreases dentin permeability.

Effect of vitamin levels and different stocking densities on performance, nutrient digestibility, and blood characteristics of growing pigs.

This study was conducted to evaluate the effects of dietary vitamin levels and stocking densities on growth performance, nutrient digestibility, and blood characteristics in growing pigs. A 2×3 factorial (two vitamin levels, three regimens of stocking densities) arrangement was utilized with 96 pigs (23.10×0.95 kg initial body weight and 63 d of age) for 36 d. The pigs were allocated to pens with different stocking density (0.64, 0.48, and 0.38 m(2)/pig, respectively). The diets used in this study were a normal diet (based on NRC) and a high level of vitamin diet (2-fold higher than normal diet). The ADG and ADFI of pigs were decreased as the stocking density increased (p = 0.03 and p = 0.01, respectively). The G/F of pigs was 5% lower in the high vitamin treatment (p = 0.03) as compared with the control treatment. The apparent total tract digestibility (ATTD) of DM and N digestibility was negatively affected by the high level of vitamin in diets (p = 0.05 and p = 0.04, respectively). Moreover, a significant and negative effect on the ATTD of N was detected in the large groups (linear, p = 0.02). Blood cortisol concentration was increased with increasing stocking density (linear, p = 0.05), and was decreased by high level of vitamin (p = 0.04) at the end of this experiment. Stocking density also caused a linear reduction in WBC concentration (p = 0.05). Our data indicated that the principal effect of stocking density was not reliant on dietary vitamin levels. In conclusion, results indicated that doubling the vitamin supplementation did not improve the growth performance of pigs in high density. However, the blood cortisol concentration was decreased but the ATTD of N digestibility was impaired by high level of vitamin diet.

Study on the inhibitory effects of Korean medicinal plants and their main compounds on the 1,1-diphenyl-2-picrylhydrazyl radical.

A 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical-generating system was used to evaluate the antioxidant properties of Korean medicinal plants that have been used widely as folk medicines for several disorders, as well as compounds isolated from them. Among the Rosaceae, Rosa rugosa and Rosa davurica showed strong DPPH radical-scavenging activity. The most effective medicinal plant from families other than Rosaceae was Cedrela sinensis, followed in order by Nelumbo nucifera, Eucommia ulmoides, Zanthoxylum piperitum, Cudrania tricuspidata and Houttuynia cordata. These results serve as a good index of the free radical-scavenging activities of Korean medicinal plants. Furthermore, the polyphenols isolated from these plants, procyanidin B-3, (+)-catechin, gallic acid, methyl gallate, quercetin, quercetin-3-O-beta-D-glucoside, quercetin-3-O-beta-galactoside, quercetin-3-O-rutinose and kaempferol, exerted strong DPPH radical-scavenging activity. These results suggest that the Korean medicinal plants and the polyphenols isolated from them that exhibited effective radical-scavenging activity may be promising agents for scavenging free radicals and treating diseases associated with excess free radicals.

Oxygen dynamics in petroleum hydrocarbon contaminated salt marsh soils: III. A rate model.

A model describing oxygen dynamics due to crude oil biodegradation under flooded conditions in saltwater wetlands was developed. The model is composed of three non-linear ordinary differential equations (ODEs) that simulate oxygen uptake, cell growth, and oil degradation simultaneously. The model equations were solved by using a stiff version of ODE solver, ODEPACK, which employs a multistep method and allows the change of step sizes and order of methods (ie., Gear's method). The results of model simulation were compared with experimental data obtained from a fully aerated microcosm study. The results of model simulation indicate that dissolved oxygen concentration in the overlying water rapidly depleted below 3 mg l(-1) unless the reaeration coefficient was higher than 2.0 day(-1). Active aerobic biodegradation of crude oil did not occur under flooded conditions because (i) dissolved oxygen is rapidly depleted, (ii) reaeration is not sufficient enough to replenish dissolved oxygen and (iii) the oil dissolution rate constant decreases over time. The model may lead to better understanding of oxygen demand for a long bioremediation period. The results of this study may be applicable for the establishment of an engineered bioremediation strategy.

Protective effects of green tea polyphenol against reactive oxygen species-induced oxidative stress in cultured rat calvarial osteoblast.

The injurious effects of reactive oxygen species on osteoblasts and the potential protective role played by green tea polyphenols (GtPP) were investigated using primarily cultured rat calvarial osteoblasts. Oxidative stress was induced in cultured osteoblasts, either by adding 100 mmol/L H2O2 or by the action of 40 U/L xanthine oxidase (XO) in the presence of xanthine (250 micromol/L). After incubation, the cellular viability, function and morphology were evaluated. Both treatments produced a significant reduction in osteoblast viability, as assessed by a two-colored fluorescence staining method combined with flow cytometric analysis and MTT assay. A significant reduction in the alkaline phosphatase activity was observed after H2O2 addition, whereas XO did not have the same effect. On the microscopic observations, the morphological changes and intracellular ultrastructural damages were remarkably induced by both treatments. The H2O2-induced alterations were prevented by pre-incubating the osteoblasts with 200 microg/ml GtPP for 1 h. When the oxidative stress was induced by XO, the cellular viability and morphology was also maintained at the same polyphenol concentration. These results demonstrate that GtPP can act as a biological antioxidant in a cell culture experimental model and protect cells from oxidative stress-induced toxicity.

Calcification of leaflets from porcine aortic valves crosslinked by ultraviolet irradiation.

Glutaraldehyde (GA)-pretreated porcine aortic valves are generally used as a bioprosthetic valve, but gradual calcification of the leaflets often occurs. It has been hypothesized that the crosslinking agent, GA, stabilizes and perhaps modifies phosphorus-rich calcifiable structures in the bioprosthetic tissue. This is supported by our findings that calcium deposition is induced rapidly in GA-pretreated leaflets in comparison with ultraviolet (UV) irradiated leaflets. After 3 days of in vitro calcification test, calcium levels were 257.6 +/- 23.5 microg/cm3 in GA-pretreated, 57.7 +/- 10.2 microg/cm3 in the control, and 108.6 +/- 7.6 microg/cm3 in 24 h UV irradiated leaflets. The calcium level in all test groups increased as time proceeds and the calcium level of GA-pretreated leaflets was significantly higher than the control and UV irradiated leaflets during test. This rapid calcium deposition on the GA-pretreated leaflets may be due to residual aldehyde groups after pretreatment. The exposure time of UV irradiation was not significantly correlated with the extent of calcification. After 14 days of the test, calcium levels in leaflets UV irradiated for 1, 2, 4, and 24 h were 502.6 +/- 12.3 microg/cm3, 547.5 +/- 34.1 microg/cm3, 564.3 +/- 26.1 microg/cm3, and 543.0 +/- 55.5 microg/cm3. In all test groups, [Ca]/[P] molar ratio decreased toward that of hydroxyapatite as the predominant mineral phase as time proceeds. This study suggests that UV irradiation can be considerable as an efficient crosslinking method to surmount the side effects induced by GA-pretreatment and may endow tissue with mechanical property.

Antioxidant flavonoids and chlorogenic acid from the leaves of Eriobotrya japonica.

The antioxidant activity of Eriobotrya japonica was determined by measuring the radical scavenging effect on DPPH (1,1-diphenyl-2-picrylhydrazyl) radical and lipid peroxidation produced when mouse liver homogenate was exposed to the air at 37 degrees C, using 2-thiobarbituric acid (TBA). The methanol extract and its fractions of Eriobotrya japonica leaves showed strong antioxidant activity. The antioxidant activity of EtOAc and n-BuOH soluble fractions were stronger than the others, and were further purified by repeated silica gel, MCl gel CHP-20P, and Sephadex LH-20 column chromatography. Antioxidant chlorogenic acid, quercetin-3-sambubioside from n-BuOH fraction, and methyl chlorogenate, kaempferol- and quercetin-3-rhamnosides, together with the inactive ursolic acid and 2 alpha-hydroxyursolic acid from EtOAc fraction were isolated. Antioxidant flavonoids and chlorogenic acid also showed prominent inhibitory activity against free radical generation in dichlorofluorescein (DCF) method.

Protective effect of Oenanthe javanica on the hepatic lipid peroxidation in bromobenzene-treated rats and its bioactive component.

To investigate the detoxification of bromobenzene-induced hepatic lipid peroxidation by Oenanthe javanica DC, the hepatic lipid peroxide level and the activities of enzymes responsible for production and removal of epoxide were studied. The level of lipid peroxide elevated by bromobenzene was significantly reduced by the methanol extract (250 mg/kg) and persicarin (5 mg/kg). The methanol extract and persicarin administered daily over 4 weeks before intoxication with bromobenzene did not affect the activities of aminopyrine N-demethylase, aniline hydroxylase, and glutathione S-transferase. Epoxide hydrolase activity was decreased significantly by bromobenzene, which was restored to the control level by pretreatment with persicarin. However, the identical pretreatment with isorhamnetin and hyperoside did not change the enzyme activity or lipid peroxide level. The results suggest that the reduction of bromobenzene-induced hepatic lipid peroxidation by O. javanica under our experimental conditions is effected through enhancing the activity of epoxide hydrolase, an enzyme removing bromobenzene epoxide. In addition, the bioactive component of this plant responsible for the detoxification of bromobenzene, at least in part, is thought to be persicarin.

Diterpenoid alkaloids from Delphinium davisii.

Three new hetisane-type diterpenoid alkaloids, davisinol (6), 18-benzoyldavisinol (7), and Davisin (9) have been isolated from Delphinium davisii Munz. and their structures established by detailed spectroscopic studies. Accurate 1H- and 13C-NMR assignments have been made for kobusine (8), a related hetisane-type alkaloid, and karakoline (5), a norditerpenoid alkaloid. The known norditerpenoid alkaloids 14-acetylperegrine (4), 6-deacetylperegrine (3), and karakoline (5) and the diterpenoid alkaloids hetisine and hetisinone were also isolated.

Acute hemodynamic improvement by thermal vasodilation in congestive heart failure.

BACKGROUND: A warm-water bath (WWB) or sauna bath (SB) has generally been considered inappropriate for patients with severe congestive heart failure (CHF). However, a comprehensive investigation of the hemodynamic effects of thermal vasodilation in CHF has not been previously undertaken. METHODS AND RESULTS: To investigate the acute hemodynamic effects of thermal vasodilation in CHF, we studied 34 patients with chronic CHF (mean age, 58 +/- 14 years). Clinical stages were New York Heart Association functional class II in 2, III in 19, and IV in 13 patients. Mean ejection fraction was 25 +/- 9%. After a Swan-Ganz catheter was inserted via the right jugular vein, the patient had a WWB for 10 minutes at 41 degrees C or an SB for 15 minutes at 60 degrees C. Blood pressure, ECG, echo-Doppler, expiration gas, and intracardiac pressures were recorded before, during, and 30 minutes after each bath. Oxygen consumption increased mildly, pulmonary arterial blood temperature increased by 1.2 degrees C, and heart rate increased by 20 to 25 beats per minute on average at the end of WWB or SB. Systolic blood pressure showed no significant change. Diastolic blood pressure decreased significantly during SB (P < .01). Cardiac and stroke indexes increased and systemic vascular resistances decreased significantly during and after WWB and SB (P < .01). Mean pulmonary artery, mean pulmonary capillary wedge, and mean right atrial pressures increased significantly during WWB (P < .05) but decreased significantly during SB (P < .05). These pressures decreased significantly from the control level after each bath (P < .01). Mitral regurgitation associated with CHF decreased during and 30 minutes after each bath. Cardiac dimensions decreased and left ventricular ejection fraction increased significantly after WWB and SB. In an additional study, plasma norepinephrine increased significantly during SB in healthy control subjects and in patients with CHF and returned to control levels by 30 minutes after SB. CONCLUSIONS: Hemodynamics improve after WWB or SB in patients with chronic CHF. This is attributable to the reduction in cardiac preload and afterload. Thus, thermal vasodilation can be applied with little risk if appropriately performed and may provide a new nonpharmacological therapy for CHF.

Two new norditerpenoid alkaloids from Delphinium elatum var. "black night".

Two new norditerpenoid alkaloids, blacknine [1] and blacknidine [2], have been isolated from the whole plants of Delphinium elatum var. "black night" along with six known alkaloids. The structures of the new alkaloids 1 and 2 were derived from their spectroscopic data (1H, 13C, DEPT, COSY, HETCOR, and selective INEPT nmr experiments). The phytochemistry of this plant variety has not been examined previously.

[Effects of hot water bath or sauna on patients with congestive heart failure: acute hemodynamic improvement by thermal vasodilation].

The acute hemodynamic effects of thermal vasodilation caused by exposure to hot water bath or sauna in chronic congestive heart failure were investigated in 32 patients (mean age 57 +/- 15 years old) with dilated cardiomyopathy (25 idiopathic and 7 ischemic). The clinical symptoms were New York Heart Association Class II in 2 patients, III in 17 and IV in 13, and the mean ejection fraction was 25 +/- 9% (9-44%). Exposure to hot water bath was for 10 minutes at 41 degrees C in a semi-sitting position, and to sauna for 15 minutes at 60 degrees C in a supine position using a special far infrared ray sauna chamber. Blood pressure, electrocardiogram, two-dimensional and Doppler echocardiograms, expiration gas, and intracardiac pressure tracings were recorded before (control), during, and 30 minutes after hot water bath or sauna. 1. The increase in oxygen consumption was only 0.3 Mets during hot water bath or sauna, and returned to the control level 30 minutes later. 2. The deep temperature in the main pulmonary artery increased by 1.0-1.2 degrees C on average at the end of hot water bath or sauna. 3. Heart rate increased significantly (p < 0.01) by 20-25/min during bathing and still increased 30 min later. 4. Systolic blood pressure did not change significantly during and after hot water bath or sauna, while, diastolic blood pressure decreased significantly during (p < 0.05) and after sauna (p < 0.01), and after hot water bath (p < 0.01).(ABSTRACT TRUNCATED AT 250 WORDS)

Impairment of the chick's grip and balance by streptomycin. A preliminary study.

Chicks were injected daily with streptomycin (400 or 1,200 mg/kg). Damage to the vestibular apparatus was accompanied by steadily impaired abilities of chicks to tighten their grips as evidenced by reduced torque and by slippage of their feet on the perch. As compensation, intoxicated chicks adjusted their footing with cautious, deliberate movements. They always stood erectly and stiffly on the perch; they were unable to roost. They also experienced difficulties in balancing. While symptoms were qualitatively similar at both dosages, their onset came earlier and with greater severity at 1,200 mg/kg. Control chicks steadily increased their torque and displayed normal balance during the course of the experiment.