Delta15N of soil N and plants in a N-saturated, subtropical forest of southern China.

We investigated the delta(15)N profile of N (extractable NH(4)(+), NO(3)(-), and organic N (EON)) in the soil of a N-saturated subtropical forest. The order of delta(15)N in the soil was EON > NH(4)(+) > NO(3)(-). Although the delta(15)N of EON had been expected to be similar to that of bulk soil N, it was higher than that of bulk soil N by 5 per thousand. The difference in delta(15)N between bulk soil N and EON (Delta(15)N(bulk-EON)) was correlated significantly with the soil C/N ratio. This correlation implies that carbon availability, which determines the balance between N assimilation and dissimilation of soil microbes, is responsible for the high delta(15)N of EON, as in the case of soil microbial biomass delta(15)N. A thorough delta(15)N survey of available N (NH(4)(+), NO(3)(-), and EON) in the soil profiles from the organic layer to 100 cm depth revealed that the delta(15)N of the available N forms did not fully overlap with the delta(15)N of plants. This mismatch in delta(15)N between that of available N and that of plants reflects apparent isotopic fractionation during N uptake by plants, emphasizing the high N availability in this N-saturated forest.

Contribution of atmospheric nitrate to stream-water nitrate in Japanese coniferous forests revealed by the oxygen isotope ratio of nitrate.

Evaluation of the openness of the nitrogen (N) cycle in forest ecosystems is important in efforts to improve forest management because the N supply often limits primary production. The use of the oxygen isotope ratio (delta(18)O) of nitrate is a promising approach to determine how effectively atmospheric nitrate can be retained in a forest ecosystem. We investigated the delta(18)O of nitrate in stream water in order to estimate the contribution of atmospheric NO(3) (-) in stream-water NO(3) (-) (f(atm)) from 26 watersheds with different stand ages (1-87 years) in Japan. The stream-water nitrate concentrations were high in young forests whereas, in contrast, old forests discharged low-nitrate stream water. These results implied a low f(atm) and a closed N cycle in older forests. However, the delta(18)O values of nitrate in stream water revealed that f(atm) values were higher in older forests than in younger forests. These results indicated that even in old forests, where the discharged N loss was small, atmospheric nitrate was not retained effectively. The steep slopes of the studied watersheds (>40 degrees ) which hinder the capturing of atmospheric nitrate by plants and microbes might be responsible for the inefficient utilization of atmospheric nitrate. Moreover, the unprocessed fraction of atmospheric nitrate in the stream-water nitrate in the forest (f(unprocessed)) was high in the young forest (78%), although f(unprocessed) was stable and low for other forests (5-13%). This high f(unprocessed) of the young forest indicated that the young forest retained neither atmospheric NO(3) (-) nor soil NO(3) (-) effectively, engendering high stream-water NO(3) (-) concentrations.

Regulation of adipose triglyceride lipase by rosiglitazone.

AIM: To elucidate the mechanism by which rosiglitazone regulates adipose triglyceride lipase (ATGL). METHODS: Male C57Bl/6 mice were treated with rosiglitazone daily (10 mg/kg body weight), and adipose tissues were weighed and preserved for mRNA and protein analysis of ATGL. In parallel, preadipocyte (3T3-L1) cells were differentiated with insulin/dexamethasone/3-isobutyl-1-methlxanthine cocktail or rosiglitazone, and ATGL levels were measured with real-time PCR, western blotting and immunohistochemistry. RESULTS: Rosiglitazone concomitantly promoted differentiation of pre-adipocytes to functional adipocytes and induced mRNA levels of ATGL. The peroxisome proliferator-activated receptor-gamma (PPARgamma) antagonist bisphenol A diglycidyl ether significantly abrogated the induction of mRNA, but not protein levels of ATGL by rosiglitazone in differentiated 3T3-L1 adipocytes. In the presence of epinephrine rosiglitazone stimulated free fatty acid release and increased diacylglycerol acyltransferase-1 (DGAT-1) mRNA suggest that ATGL and DGAT-1 may be cooperatively involved in rosiglitazone-stimulated triglyceride hydrolysis and fatty acid re-esterification in 3T3-L1 adipocytes. Treatment of 3T3-L1 adipocytes with rosiglitazone or insulin did not appear to alter localization of ATGL staining surrounding lipid droplets. Finally, we found that rosiglitazone increased ATGL mRNA levels in 3T3-L1 adipocytes in the presence of cycloheximide, an inhibitor of protein synthesis, suggesting that rosiglitazone regulation of ATGL occurs at the transcriptional level. CONCLUSIONS: Rosiglitazone directly regulates transcription of ATGL, likely through a PPARgamma-mediated mechanism.

The potential of NO3--N utilization by a woody shrub species Lindera triloba: a cultivation test to estimate the saturation point of soil NO3--N for plants.

Responses of seedlings of a shrub species, Lindera triloba, grown in perlite culture medium, to nitrate (NO3--N) supply were investigated to estimate the saturating point of available NO3--N for plant utilization. NO3--N concentration and nitrate reductase activity (NRA) in leaves and roots were used as indicators of NO3--N uptake and assimilation by L. triloba. Root NRA increased with NO3--N supply when concentrations were low and reached a plateau at high NO3--N concentrations. On the other hand, root NO3--N concentration increased linearly with NO3--N supply; therefore, it is suggested that NO3--N uptake did not limit NO3--N assimilation by L. triloba. In contrast, leaf NRA and leaf NO3--N concentration were low and were not influenced by NO3--N supply. This may be caused by the lack of transport of NO3--N from roots to leaves. The NO3--N retained in perlite was compared with NO3--N pool sizes in soils from a forest where L. triloba occurs naturally to estimate the level of NO3--N availability to plants in the forest soil. The maximum NO3--N pool size in the forest soil was comparable to concentrations at which root NRA reached a plateau in perlite cultures. These results indicate that soil NO3--N availability is below the saturation point for NO3--N uptake by L. triloba, and it is the limiting factor of NO3--N utilization by L. triloba under field conditions in which this species naturally occurs.

Dietary manipulations of body fat-reducing potential of conjugated linoleic acid in rats.

To study whether the body fat-reducing potential of conjugated linoleic acid (CLA) could be increased through dietary manipulations, the effects of the combination of CLA with different proteins, fats, and sesamin were examined in rats. Male rats were fed diets containing 1% CLA or linoleic acid (LA) in combination with different proteins (20% of casein or soybean protein), fats (7% perilla oil or soybean oil) and 0.2% sesamin (SES) for 3 or 4 weeks. When the dietary fat source was soybean oil, CLA, as compared with LA, significantly reduced weights of epididymal and perirenal adipose tissues, irrespective of the dietary protein sources. However, the highest reducing effect was shown when soybean protein was given as a protein source. SES stimulated the reduction of epididymal and perirenal adipose tissue weights in both protein diets. In contrast, CLA increased the weight of brown adipose tissue, and SES further increased it in combination with soybean oil but not with perilla oil. No effect of dietary manipulation was observed on serum leptin and TNF-alpha levels. Thus, the body fat-reducing potential of CLA can be increased by an appropriate combination with food factors that may stimulate fatty acid beta-oxidation.

Cholesterol supplementation attenuates the hypocholesterolemic effect of rice bran oil in rats.

Rice bran oil (RBO), when blended with safflower oil (SFO) at the ratio of 7 to 3, has been shown to lower serum cholesterol in humans consuming cholesterol. The mechanism as to how this oil blend exerts its effect is not yet clear. This study examined the effect of cholesterol supplementation on the cholesterol-lowering ability of different RBO/SFO blends. Male Sprague Dawley rats (4 wk old) were fed purified diets containing 10% fat with or without the addition of 0.5% cholesterol for 3 wk. The fat was either SFO or RBO alone, or the mixture of these two oils at the ratio of 7: 3 (7S/3R), 5:5 (5S/5R), or 3:7 (3S/7R). Without cholesterol supplementation, there were no significant differences in the serum and liver total cholesterol levels among different dietary fats. However, the HDL cholesterol level of rats fed the RBO-containing diets (especially in rats fed the 3S/7R diet) was higher than that of rats fed the diet containing SFO alone. This resulted in an increase in the ratio of HDL/total cholesterol-a desirable outcome. Supplementation of the diets with 0.5% cholesterol significantly increased the cholesterol level in both the serum and the liver. Increasing the proportion of RBO in the diet further raised the total cholesterol level in the serum whereas it reduced liver cholesterol. Then, the specific effect of the 3S/7R mixture on the ratio of HDL/total cholesterol disappeared. These findings suggest that cholesterol supplemented at the level of 0.5% in this study masked the cholesterol-lowering effect of RBO. Smaller percentages of polyunsaturated fatty acid (i.e., 18:2n-6) in the RBO-containing diets than in the SFO diet might have reduced their ability to dispose the circulating serum cholesterol into the liver.

Modulation of cholesterol concentration in Caco-2 cells by incubation with different n-6 fatty acids.

Incorporation of exogenous cholesterol was compared in human adenocarcinoma colon cells (Caco-2) after incubation with 100 microM of either linoleic acid (LA, 18:2n-6), gamma-linolenic acid (GLA, 18:3n-6), arachidonic acid (AA, 20:4n-6) or adrenic acid (or n-6 docosatetraenoic acid, DTA, 22:4n-6). In both cells 7 days after seeding and 14 days after confluency, incubation with LA significantly raised the proportion of 18:2n-6 but not its long-chain metabolites in cellular phospholipid. Incubation with GLA increased the levels of 18:3n-6, 20:3n-6, and 20:4n-6. Incubation with AA increased the levels of 20:4n-6 and 22:4n-6, and incubation with DTA increased the levels of 22:4n-6 as well as its retro-conversion metabolite, 20:4n-6. A subsequent addition of cholesterol (180 microM) to the medium significantly raised the cellular cholesterol level but less so in the cells 7 days after seeding incubated with GLA. The increase in cellular cholesterol level was generally greater in the cells of 7 days after seeding, particularly those incubated with long-chain highly unsaturated n-6 fatty acids, than in those of 14 days after confluency. These findings suggest that the cell growth and the extent of unsaturation in cell membrane phospholipid fatty acids modulate the incorporation of the exogenous cholesterol into the Caco-2 cells.

Post-operative symptoms and healing after endodontic treatment of infected teeth using pulsed Nd:YAG laser.

Forty-four teeth in 38 patients, diagnosed with chronic apical periodontitis, were endodontically treated. Root canals were shaped using a step-back technique with 5% NaOCl and 3% H2O2 as irrigants. In half of the teeth the canal terminus was irradiated with pulsed Nd:YAG laser (1 W, 15 pps, 1 s). All canals were then obturated with laterally condensed gutta-percha points and sealer, and final radiographs were obtained. Occurrence of spontaneous pain was recorded 1 day after treatment. Percussion pain was recorded after 1 week, and then at 3 and 6 months after treatment. Radiographic follow-up was performed at 3 and 6 months. Percussion pain was significantly less (P < 0.05) in the laser-treated group than in the control group, both 1 week and 3 months after treatment. Other differences between the groups were not significant. These results suggested that the clinical application of pulsed Nd:YAG laser might be advantageous for the treatment of infected root canals.

A histopathological study of the effects of pulsed Nd:YAG laser irradiation on infected root canals in dogs.

The effects of pulsed Nd:YAG laser irradiation during root canal treatment of infected teeth were investigated histopathologically in dogs. One hundred and eight teeth with a single root in 15 healthy adult beagle strain dogs were used in this study. After inducing infected teeth, each root canal was shaped with up to at least a #40 K-file; then, after coating with black ink, the canal was irradiated using the following parameters: 1 W, 30 pps, for 1 and 2 s, and 2 W, 30 pps, for 2 s. The degree of inflammation of the periapical regions at 2, 4, and 8 wk was examined histopathologically by light microscopy. Inflammation of the periapical regions in the laser-treated groups was significantly less than that in the control group at 4 and 8 wk (p < 0.05). These results suggest that pulsed Nd:YAG laser is useful for root canal treatment of infected teeth, if appropriate parameters are selected.

A clinical study on the effects of pulsed Nd:YAG laser irradiation at root canals immediately after pulpectomy and shaping.

OBJECTIVE: The effects of pulsed Nd:YAG laser irradiation for the treatment of root canals immediately after pulpectomy and shaping with regard to improvement of symptoms were evaluated clinically. SUMMARY BACKGROUND DATA: The effects of pulsed Nd:YAG laser irradiation for the treatment of root canals were experimentally investigated using an animal model, but not under clinical conditions. MATERIALS AND METHODS: Thirty-eight teeth from 29 patients were diagnosed as normal or irreversible pulpitis were used. After extirpation of pulp and shaping using a step-back technique and cleansing with 5% sodium hypochlorite (NaOCl) and 3% hydrogen peroxide (H2O2), laser irradiation was applied at 1 W and 15 pps for 1 second in 23 teeth (laser-treated group). Root canals were then obturated with gutta-percha points and sealer by the lateral condensation method. The condition of the obturations was confirmed radiographically. The control teeth were treated in the same way, but without laser irradiation. Occurrence of spontaneous pain was recorded 1 day after treatment and occurrence of percussion pain was recorded 1, 2, and 3 weeks after treatment. RESULTS: Effective ratio, which was the ratio of the number of "none" teeth to total teeth number in the laser-treated group, was higher than that in the control group, but there was no significant difference between 2 groups with regard to spontaneous and percussion pain (p > 0.05). CONCLUSION: These results suggest that the clinical application of pulsed Nd:YAG laser might be advantageous for the one-visit treatment of root canals immediately after pulpectomy shaping and to reduce postoperative pain.

Health benefits of rice bran oil.

Although scientific evidence is relatively limited, rice bran oil (RBO) is tenaciously believed to be a healthy vegetable oil in Asian countries. It exerts hypocholesterolemic activity in relation to more commonly used vegetable oils and is characterized by a relatively high content of non-fatty acid components, some of which are known to have beneficial health effects. Components specific for RBO such as gamma-oryzanol and tocotrienols could participate in its hypocholesterolemic effects. In addition, blending RBO with safflower oil, but not with sunflower oil, may magnify the hypocholesterolemic efficacy. This observation is of particular interest with regard to dietary intervention with RBO. The possible mechanism underlying this effect may at least in part be related to the specific triglyceride structure of safflower oil, differing from that of sunflower oil.

Pulsed Nd:YAG laser application to one-visit treatment of infected root canals in dogs: a histopathological study.

OBJECTIVE: In this study, the effects of pulsed Nd:YAG laser irradiation during root canal treatment of infected teeth were investigated histopathologically in dogs. SUMMARY BACKGROUND DATA: Effects of Nd:YAG laser on infected root canal treatment have not been reported in vivo. METHODS: One hundred thirty-five teeth with a single root, including incisors and premolars, in 15 healthy adult beagle-strain dogs were used in this study. After inducing infection in the teeth, each root canal was shaped with at least a # 40 K-file, then the canal was irradiated using the following parameters: 1 W, 30 pps for 1 and 2 sec; 2 W, 30 pps for 1 and 2 sec. Efficacy of debris removal and evaporation on the root canal walls at 2 weeks, and the degree of inflammation of the periapical region at 2, 4, and 8 weeks after laser irradiation were examined histopathologically by light microscopy. RESULTS: Effective debris removal was observed in all cases of the laser-treated groups. No evaporation was observed except at the conditions of 2 W for 2 sec. Inflammation of periapical region in the laser-treated groups was similar in the control group at 2 weeks, but was significantly less than that in the control group at 8 weeks (p < 0.05). CONCLUSION: These results suggest that pulsed Nd:YAG laser is useful for one-visit root canal treatment of infected teeth in dogs, if appropriate parameters are selected, and this is a potential therapy for human apical lesions of teeth.

A histopathological study of the morphological changes at the apical seat and in the periapical region after irradiation with a pulsed Nd:YAG laser.

In this study, the effects of pulsed Nd:YAG laser irradiation on the apical seat and periapical region of instrumented root canals were investigated. One hundred and ninety-two teeth, including incisors and molars (239 root canals), in 18 healthy adult beagle dogs were used. After extirpation of the pulp under anaesthesia, each root canal was shaped up to at least a size 40 K-file, and then laser irradiated using the following parameters: 1 W, 15 pps for 2, 3, and 4 s; 2 W, 15 pps for 2 s. The effectiveness of debris removal and carbonization on the root canal walls immediately laser irradiation were examined and inflammation of the periapical region at 1, 2, and 4 weeks after laser irradiation was examined using light microscopy. Effective debris removal was observed in most cases in the laser-treated groups and was dependent on irradiated energy. Carbonization was observed in the irradiated root canals except at the parameter of 1 W for 2 s. Periapical inflammation was observed at 1 W for 2 s to the same extent as in the control group (P > 0.01). These results suggest that the pulsed Nd:YAG laser is useful for debris removal with no adverse effects immediately after extirpation of the pulp if appropriate parameters are selected.

The effect of pulpotomy using CO2 and Nd:YAG lasers on dental pulp tissue.

The histological response of the dental pulp after laser irradiation was studied. After pulpotomy was performed in the premolar and molar teeth of dogs, the exposed pulp tissue at the root canal opening was lased using either a CO2 or Nd:YAG laser. The laser parameters were 2 W, 10 ms, 5 times per second for 1, 2 and 3 s for CO2 laser and 2 W, 20 pulses per second for 1, 2 and 3 s for the Nd:YAG laser. Observations were made 30 and 45 days after treatment. The results revealed that laser irradiation caused carbonization, necrosis, infiltration of inflammation cells, oedema and haemorrhage in the pulp tissue. Under the conditions of this experiment, there was little histological evidence of repair to the treated pulp with a newly formed dentine barrier, which was in contrast to the control samples treated with a calcium hydroxide-containing cement (Dycal).

Distribution of plasma phosphatidylcholine molecular species in rabbits fed fish oil is modulated by dietary n-6 fatty acids.

The present study examined the distribution of plasma phosphatidylcholine (PC) molecular species in rabbits fed a chow diet supplemented with fish oil (FO) in combination with either hydrogenated coconut oil or the n-6 fatty acid-rich evening primrose oil (EPO) for 4 weeks. Significant proportions of plasma PC molecular species contained long-chain n-3 fatty acids. Addition of EPO to the FO supplemented diet increased the incorporation of n-6 fatty acids into plasma PC molecules; it also raised the proportions of 16:0-18:2, n-6, 18:1-18:2, n-6, 18:2, n-6-18:2, n-6, and 16:0-20:4, n-6. The increase of n-6 fatty acid-containing PC was at the expense of n-3 fatty acid containing PC species. However, feeding n-6 fatty acids did not affect the distribution of PC molecular species based on total carbon chain length. The most interesting observation was that dietary suplementation with EPO, raised the ratio of 22:6, n-3-containing to 20:5, n-3-containing molecular species, suggesting an enhanced conversion of 20:5, n-3 to 22:6, n-3.

Effect of calcium hydroxide paste dressing on uninstrumented root canal wall.

Dissolution of soft tissues on the uninstrumented root canal wall by an intracanal dressing of calcium hydroxide paste was examined by scanning electron microscopy (SEM). The roots of human extracted teeth were split longitudinally into halves, and the pulp tissue was removed by forceps. Then, the halves were divided randomly into three groups. Specimens in the control group were given no treatment. Specimens of the 1-wk group were stored for 7 days in calcium hydroxide-saline paste under the condition of 100% humidity and 37 degrees C. Specimens of the 4-wk group were stored for 28 days in the same manner, but washed and replaced into new paste every week. Root canal walls of these specimens were compared under a scanning electron microscope. A layer of odontoblasts was observed in the control group, whereas it had disappeared completely in the 1- and 4-wk groups. However, the predentin remained almost intact in the 1-wk group, and although the surface was eroded, in the 4-wk group. It was found that calcium hydroxide paste dressing could dissolve the odontoblastic cell layer, but had little effect on predentin.

N-3 and n-6 fatty acid metabolism in undifferentiated and differentiated human intestine cell line (Caco-2).

Metabolism of n-6 and n-3 fatty acids in the undifferentiated and differentiated human adenocarcinoma colon cell line (Caco-2) was studied. In cells incubated with either 18:2n-6 or 18:3n-3, no significant amounts of long chain n-6 and n-3 metabolites were found. Incubation with either 18:3n-6 or 18:4n-3 raised significantly the levels of 20:3n-6 and 20:4n-3, respectively. In the undifferentiated cells, significant proportions of 20:3n-6 and 20:4n-3 were further delta 5-desaturated to form 20:4n-6 and 20:5n-3, respectively. Incubation with either 20:4n-6 or 20:5n-3 raised the levels of their direct elongation products, 22:4n-6 and 22:5n-3, respectively. Incubation with 22:4n-6 or 22:5n-3 increased the levels of 20:4n-6 and 20:5n-6. These results suggest that delta 6-desaturation in the Caco-2 cells is less active in comparison with elongation, delta 5-desaturation and retro-conversion. These enzymes were modulated by the state of differentiation, and appeared to be non-specific to n-3 and n-6 fatty acids. When cells were incubated with 18:3n-6 and 18:4n-3 concomitantly, the levels of incorporation of total n-6 fatty acids into cellular lipids were greater than those of the n-3 fatty acids, whereas the ratios of 20+22 carbon metabolites to 18-carbon precursor favored n-3 over n-6 fatty acids. These results suggest that n-3 and n-6 fatty acids were not metabolized identically in Caco-2 cells.

Effects of dietary protein and cholesterol on phosphatidylcholine and phosphatidylethanolamine molecular species in mouse liver.

The present study examined the effects of two atherogenic factors, animal protein and cholesterol, on the distribution of fatty acids and the molecular species of major liver phospholipids in mice. Weanling mice were fed a semisynthetic diet supplemented with either casein or soy protein (20%, w/w) in the presence or absence of 0.5% cholesterol for 4 wk. Results from mouse liver showed that animal protein and, more so, dietary cholesterol modified the fatty acid profiles of the phospholipids. Animal protein had no significant effect on the concentration of lipids, but it altered the relative distribution and fatty acid profiles of the phospholipids, phosphatidylcholine and phosphatidylethanolamine. Dietary cholesterol, on the other hand, significantly increased the concentration of liver lipids, but it did not alter the relative distribution of phosphatidylcholine and phosphatidylethanolamine. In cholesterol-fed mice, the proportions of molecular species containing 18:2n-6 were increased, whereas those containing 20:4n-6 were decreased, indicating that dietary cholesterol suppressed linoleic acid metabolism. Since cholesterol feeding selectively decreased the ratio of 18:0/20:4n-6 in phosphatidylcholine, whereas it increased the 18:0/18:2n-6 ratio in phosphatidylethanolamine, this finding suggests that dietary cholesterol may affect the incorporation of fatty acids but not the rate of synthesis of phosphatidylcholine and phosphatidylethanolamine.

Effects of dietary proteins on linoleic acid desaturation and membrane fluidity in rat liver microsomes.

The effect of dietary protein, casein (CAS) and soybean protein (SOY), on linoleic acid desaturation in liver microsomes was studied in rats. The activity of delta 6 desaturase in total and rough endoplasmic reticula (ER and RER) was significantly higher in the CAS group than in the SOY group. In ER and smooth endoplasmic reticulum, the steady-state fluorescence anisotropy of 1,6-diphenyl-1,3,5-hexatriene, when incorporated into the membrane, was decreased in the SOY group and accompanied by a reduction in the cholesterol/phospholipid (CHOL/PL) ratio, consistent with an increase in membrane fluidity. In a separate study, the effect of varying dietary proteins, CAS, milk whey protein, egg albumin, SOY, potato protein and wheat gluten, on the relationship between the delta 6 desaturase activity and microsomal membrane fluidity was also examined. The results indicated that the dietary protein-dependent change in the liver microsomal CHOL/PL ratio affected membrane fluidity, and subsequently the activity of delta 6 desaturase in liver microsomes. However, since dietary protein influenced the delta 6 desaturase activity in RER without influencing membrane fluidity, it is possible that some regulation might have taken place at the level of enzyme synthesis.