A DFT and multi-configurational perturbation theory study on O2 binding to a model heme compound via the spin-change barrier.

Dioxygen binding to a model heme compound via intersystem crossing (ISC) was investigated with a multi-state multi-configurational self-consistent field method with second-order perturbation theory (MS-CASPT2) and density functional theory (DFT) calculations. In elongated Fe-O distances, the energy levels of the S0 and T1 states are separated, which decreases the probability of intersystem crossing in these structures. At the DFT(B97D) level of calculation, the Fe-O distances of the S0 and T1 states were 1.91 and 2.92 Å, respectively. The minimum energy intersystem crossing point (MEISCP) was located as a transition state at a Fe-O distance of 2.17 Å with an energy barrier of 1.0 kcal mol(-1) from the T1 minimum. The result was verified with MS-CASPT2 calculations including the spin-orbit interaction which also showed the intersystem crossing point at a Fe-O distance of 2.05 Å. An energy decomposition analysis on the reaction coordinate showed the important contribution of the ring-shrinking mode of the porphyrin ring, indicating that the reaction coordinates which control the relative energy level of the spin-states play a key role in intersystem crossing.

Antitumour-promoting and cytotoxic constituents of etlingera elatior.

Phytochemical studies on rhizome of Etlingera elatior have resulted in the isolation of 1,7-bis(4-hydroxyphenyl)-2,4,6-heptatrienone (1), demethoxycurcumin (2), 1,7-bis(4-hydroxyphenyl)-1,4,6-heptatrien-3-one (3), 16-hydroxylabda-8(17),11,13-trien-16,15-olide (4), stigmast-4-en-3-one (5), stigmast-4-ene-3,6-dione (6), stigmast-4-en-6b-ol-3-one (7), 5α,8α-epidioxyergosta-6,22-dien-3ß-ol (8). 1 and 4 were new compounds. Compounds 5 and 7 displayed high antitumour-promoting activity. Ethyl acetate extract showed a very significant cytotoxic activity against CEM-SS and MCF-7 cell lines (4 µg/ml and 6.25 µg/ml respectively). The antitumour-promoting activity was determined by EBV-EA assay and cytotoxic activity was determined by MTT assay.

Treatment of supracondylar fracture of the humerus in children by skeletal traction in a brace.

In 1980, we developed a specially designed brace for treating supracondylar fractures of the humerus in children, along with an easy and safe technique of reduction by skeletal traction. This method, which takes into consideration only the medial tilting and anterior angulation of the distal fragment, achieves complete reduction, ignoring any lateral, posterior and minor rotational displacements of the fragment. Skeletal traction is applied through a screw inserted into the olecranon and the angulation at the fracture site is reduced regardless of the anatomical position without manipulation. We treated 193 children with displaced supracondylar fractures of the humerus using this method between 1980 and 2001. Only four children (2%) developed cubitus varus. The majority obtained an excellent range of movement at the elbow; one had a 25 degree limitation of flexion. This technique is an effective and easy method of treating supracondylar fractures of the humerus in children.

Comprehensive expression analysis of a rat depression model.

Herein we report on a large-scale analysis of gene expression in the 'learned helplessness' (LH) rat model of human depression, using DNA microarrays. We compared gene expression in the frontal cortex (FC) and hippocampus (HPC) of untreated controls, and LH rats treated with saline (LH-S), imipramine or fluoxetine. A total of 34 and 48 transcripts were differentially expressed in the FC and HPC, respectively, between control and LH-S groups. Unexpectedly, only genes for NADH dehydrogenase and zinc transporter were altered in both the FC and HPC, suggesting limited overlap in the molecular processes from specific areas of the brain. Principal component analysis revealed that sets of upregulated metabolic enzyme genes in the FC and downregulated genes for signal transduction in the HPC can distinguish clearly between depressed and control animals, as well as explain the responsiveness to antidepressants. This comprehensive data could help to unravel the complex genetic predispositions involved in human depression.

Apianane terpenoids from Salvia officinalis.

Three apianane terpenoids, rel-(5S, 6S, 7S, 10R, 12S, 13R)-7-hydroxyapiana-8,14-diene-11,16-dion-(22,6)-olide, rel-(5S, 6S, 7R, 10R, 12S, 13R)-7-hydroxyapiana-8,14-diene-11,16-dion-(22,6)-olide and rel-(5S, 6S, 7S, 10R, 12R, 13S)-7-hydroxyapiana-8,14-diene-11,16-dion-(22,6)-olide were isolated from the leaves of Salvia officinalis, together with 15 known compounds, salvigenin, lupeol, beta-sitosterol, stigmasterol, physcion, carnosol, rosmadial, rosmanol, epirosmanol, isorosmanol, columbaridione, atuntzensin A, miltirone, carnosic acid and 12-O-methyl carnosic acid. Their structures were established on the basis of spectral evidence.

Furanosesquiterpenoids of Commiphora myrrha.

An investigation on the gum exudates of Commiphora myrrha has led to the isolation of six sesquiterpenoids. On the basis of spectroscopic data interpretation, they were determined as two new furanosesquiterpenoids, rel-1S,2S-epoxy-4R-furanogermacr-10(15)-en-6-one (1) and rel-2R-methyl-5S-acetoxy-4R-furanogermacr-1(10)Z-en-6-one (2), and four known furanosesquiterpenoids, rel-3R-methoxy-4S-furanogermacra-1E,10(15)-dien-6-one (3), rel-2R-methoxy-4R-furanogermacr-1(10)E-en-6-one (4), furanogermacra-1(10)Z,4Z-dien-6-one, and curzerenone [6,7-dihydro-5beta-isopropenyl-3,6beta-dimethyl-6-vinylbenzofuran-4(5H)-one]. This is the first report of the relative stereochemistry for the known compounds 3 and 4. Compound 1 exhibited weak cytotoxic activity against a MCF-7 breast tumor cell line in a clonogenic assay, while the other five compounds were inactive in this assay.

Antioxidative activity of carbazoles from Murraya koenigii leaves.

The antioxidative properties of the leaves extracts of Murraya koenigii using different solvents were evaluated based on the oil stability index (OSI) together with their radical scavenging ability against 1-1-diphenyl-2-picrylhydrazyl (DPPH). The methylene chloride (CH(2)Cl(2)) extract and the ethyl acetate (EtOAc) soluble fraction of the 70% acetone extract significantly prolonged the OSI values comparable to those of alpha-tocopherol and BHT. Five carbazole alkaloids were isolated from the CH(2)Cl(2) extract and their structures were identified to be euchrestine B (1), bismurrayafoline E (2), mahanine (3), mahanimbicine (4), and mahanimbine (5) based on (1)H and (13)C NMR and mass (MS) spectral data. The OSI value of carbazoles at 110 degrees C decreased in the order 1 and 3 > alpha-tocopherol > BHT > 2 > 4, 5 and control. It is assumed that compounds 1 and 3 contributed to the high OSI value of the CH(2)Cl(2) extract of M. koenigii. The DPPH radical scavenging activity for these carbazoles was in the order ascorbic acid > 2 > 1, 3 and alpha-tocopherol > BHT > 4 and 5.

Antibacterial compounds of licorice against upper airway respiratory tract pathogens.

The antibacterial activity of compounds obtained from licorice was measured against upper airway respiratory tract bacteria such as Streptococcus pyogenes, Haemophilus influenzae and Moraxella catarrhalis. Among the tested compounds, licoricidin exhibited the highest activity against all tested microorganisms with an MIC of 12.5 microg/mL. Three coumarin derivatives, glycyrol, glycyrin and glycycoumarin also showed antibacterial activity.

1,1-Diphenyl-2-picrylhydrazyl radical-scavenging active compounds from greater cardamom (Amomum subulatum Roxb.).

Constituents of the fruits of greater cardamom (Amomum subulatum) were fractionated into three fractions, the dichloromethane extract, and the ethyl acetate-soluble and water-soluble fractions of the 70% aqueous acetone extract. The ethyl acetate-soluble fraction showed a high radical-scavenging activity against 1,1-diphenyl-2-picrylhydrazyl (DPPH). Four compounds were isolated from the ethyl acetate-soluble fraction, and their structures were ascribed to protocatechualdehyde (1), protocatechuic acid (2), 1,7-bis(3,4-dihydroxyphenyl)hepta-4E,6E-dien-3-one (3) and 2,3,7-trihydroxy-5-(3,4-dihydroxy-E-styryl)-6,7,8,9-tetrahydro-5H-benzocycloheptene (4) on the basis of spectroscopic evidence. This is the first isolation of these compounds from greater cardamom. In particular, 4 was a new type of cyclic diarylheptanoid. DPPH radical-scavenging activity of these compounds was measured by colorimetric analysis. Compounds 1 and 3 showed stronger activity than such natural antioxidants as alpha-tocopherol and L-ascorbic acid. Compounds 2 and 4 were comparable to alpha-tocopherol and L-ascorbic acid.

Various cooking methods and the flavonoid content in onion.

Onion is a major source of flavonoids and is cooked in various ways in the world. The major flavonoids in onion are two quercetin glycosides, quercetin 4'-O-beta-glucoside (Q4'G) and quercetin 3,4'-O-beta-diglucosides (Q3,4'G), which are recognized as bioactive substances that are good for our health. We have investigated the effect of cooking procedures on the content of antioxidants. We selected quercetin conjugates, total phenol compounds, and ascorbic acid to estimate the amount of flavonoid ingestion from onion. We examined the following cooking methods: boiling, frying with oil and butter, and microwave cooking. Various cooking methods do not consider the degradation of quercetin conjugates when cooking onion. Microwave cooking without water better retains flavonoids and ascorbic acid. Frying does not affect flavonoid intake. The boiling of onion leads to about 30% loss of quercetin glycosides, which transfers to the boiling water. At that time, the effect of additives on the quercetin conjugates is different according to the compounds. The hydrolysis of quercetin glycosides for daily cooking might occur with the addition of seasonings such as glutamic acid. Additional ferrous ions accelerated the loss of flavonoids.

Ecdysteroids of quinoa seeds (Chenopodium quinoa Willd.).

Quinoa (Chenopodium quinoa) is a hardy and nutritious Latin American pseudo-cereal. Studies on the seeds led to the isolation of five ecdysteroids using column chromatography. Their structures were determined as ecdysterone, makisterone A, 24-epi-makisterone A, 24(28)-dehydromakisterone A, and 20,26-dihydroxyecdysone by spectroscopic methods. This study demonstrates that quinoa seeds are a source of ecdysteroids, which were reported to be molting hormones in insects.

Reduced hepatic triglyceride secretion in rats fed docosahexaenoic acid-rich fish oil suppresses postprandial hypertriglyceridemia.

To evaluate the mechanisms of suppression of postprandial hypertriglyceridemia by fish oil rich in docosahexaenoic acid, the effect on the intestinal absorption of triglyceride, activities of lipoprotein lipase (LPL) and hepatic triglyceride lipase (HTGL) and metabolism of chylomicrons (CM) and CM remnants were compared with that of safflower oil in Sprague-Dawley rats in a series of studies. The feeding of fish oil for 3 wk suppressed postprandial hypertriglyceridemia (study 1). Dietary fish oil did not alter the rate of lymphatic absorption of triglyceride (study 2). The activities of LPL and HTGL were measured at 5 h after the beginning of feeding, when serum triglyceride concentrations were highest in both dietary groups. The activities of LPL in adipose tissue and heart were greater (P < 0.05) and those of HTGL were lower (P < 0.05) in the rats fed fish oil (study 3). In contrast, there were no differences in the activities of LPL and HTGL in postheparin plasma between the fish and safflower oil groups (study 4). The clearance rates of CM and CM remnants were measured by injecting intravenously CM collected from rats fed safflower or fish oils with [14C]triolein and [3H]cholesterol (study 5). Dietary oil did not influence the half-lives of CM or CM remnants. The secretion of triglyceride from the liver of rats injected with Triton WR-1339 was lower (P < 0.05) in the rats fed docosahexaenoic acid, a major component of fish oil, than those fed linoleic acid, a major component of safflower oil (study 6). These observations strongly support the hypothesis that in rats, the principal cause of the suppression of postprandial hypertriglyceridemia by fish oil is the depression of triglyceride secretion from the liver.

New constituents of the leaves of Alpinia flabellata.

Two new compounds were isolated from the leaves of Alpinia flabellata. The structures of these compounds were determined by a combination of NMR techniques and HREIMS as 4-hydroxy-2-(2,4,5-trimethoxyphenyl)-2E-butenal (1) and rel-labd-12-en-15(16)-olid-7-one-8R-spiro-1'-[2S-(2,4,5-trimethoxyphenyl)-3-cyclohexene] (2).

Measurement of ethylene emission from Japanese red pine (Pinus densiflora) under field conditions in NOx-polluted areas.

Emission of ethylene from the needles of Japanese red pine, Pinus densiflora, was measured in air-polluted areas in Hiroshima, Japan. We applied a suitable protocol to determine the rate of ethylene emission from the excised needles. The influence of excision of needles on ethylene emission was not detected during the first 4 h of incubation at 20 degrees C. Ethylene emissions were low in the unpolluted (clean) areas regardless of the altitude or season. The emission of stress ethylene increased with the atmospheric NO2 concentration, suggesting that atmospheric NOx or related substances induced the higher ethylene emission in the polluted areas (near urban and industrial areas). In all cases, 1-year-old needles emitted significantly larger amounts of ethylene than the current needles. Ethylene emission did not increase evenly in the polluted areas, but the frequency of trees emitting high ethylene increased. Therefore, threshold rates for the baseline ethylene emission were proposed.

Phenylbutanoid dimers from the leaves of Alpinia flabellata.

Three phenylbutanoid dimers, cis- and trans-1-(2,4,5-trimethoxy-E-styryl)-2-(2,4,5-trimethoxy-Z-styryl)cyclobutane and 1,2-bis(2,4,5-trimethoxy-Z-styryl)- cyclobutane, were isolated from the leaves of Alpinia flabellata Ridley, together with three known compounds (2,4,5-trimethoxybenzaldehyde, 2,4,5-trimethoxycinnamaldehyde and 3,5-dihydroxy-7,4'-dimethoxyflavone). The structures of these compounds were determined by spectroscopic analysis.

Identification, quantitative determination, and antioxidative activities of chlorogenic acid isomers in prune (Prunus domestica L. ).

Neochlorogenic acid (3-CQA) and cryptochlorogenic acid (4-CQA), isolated from prune (Prunus domestica L.), were identified by NMR and MS analyses. In addition, the quantity of chlorogenic acid isomers in prune were measured by HPLC. These isomers, 3-CQA, 4-CQA, and chlorogenic acid (5-CQA), were contained in the ratio 78.7:18. 4:3.9, respectively. 4-CQA was identified and quantified in prune for the first time, and relatively high amounts of this isomer were characteristic. Antioxidative activities of the chlorogenic acid isomers, such as scavenging activity on superoxide anion radicals and inhibitory effect against oxidation of methyl linoleate, were also evaluated. Each isomer showed antioxidative activities which were almost the same.

Role of cytosolic phospholipase A2 in the production of lipid mediators and histamine release in mouse bone-marrow-derived mast cells.

Cytosolic phospholipase A(2) (cPLA(2)) plays a critical role in mast-cell-related allergic responses [Uozumi, Kume, Nagase, Nakatani, Ishii, Tashiro, Komagata, Maki, Ikuta, Ouchi et al. (1997) Nature (London) 390, 618-622]. Bone-marrow-derived mast cells from mice lacking cPLA(2) (cPLA(-/-)(2) mice) were used in order to better define the role of cPLA(2) in the maturation and degranulation of such cells. Cross-linking of high-affinity receptors for IgE (FcepsilonRI) on cells from cPLA(-/-)(2) mice led to the release of negligible amounts of arachidonic acid or its metabolites, the cysteinyl leukotrienes and prostaglandin D(2), indicating an essential role for cPLA(2) in the production of these allergic and pro-inflammatory lipid mediators. In addition, the histamine content of the mast cells and its release from the cells were reduced to 60%. While these results are in agreement with a reduced anaphylactic phenotype of cPLA(-/-)(2) mice, the ratios of release of histamine and beta-hexosaminidase were, paradoxically, significantly higher for cells from cPLA(-/-)(2) mice than for those from wild-type mice. Consistently, IgE-induced calcium influx in mast cells was greater and more prolonged in cells from cPLA(-/-)(2) mice than in those from wild-type mice. Thus the loss of cPLA(2) not only diminishes the release of lipid mediators, but also alters degranulation. While the overall effect is still a decrease in the release of mast cell mediators, explaining the in vivo findings, the present study proposes a novel link between cPLA(2) and the degranulation machinery.

Novel glycosides from noni (Morinda citrifolia).

Three new glycosides were isolated from the fruits of noni (Morinda citrifolia). Their structures were determined to be 6-O-(beta-D-glucopyranosyl)-1-O-octanoyl-beta-D-glucopyranose (1), 6-O-(beta-D-glucopyranosyl)-1-O-hexanoyl-beta-D-glucopyranose (2), and 3-methylbut-3-enyl 6-O-beta-D-glucopyranosyl-beta-D-glucopyranoside (3) using MS and NMR methods.

Screening of Zingiberaceae extracts for antimicrobial and antioxidant activities.

Dichloromethane and methanol extracts of 13 Zingiberaceae species from the Alpinia, Costus and Zingiber genera were screened for antimicrobial and antioxidant activities. The antimicrobial activity of most of the extracts was antibacterial with only the methanol extract of Costus discolor showing very potent antifungal activity against only Aspergillus ochraceous (MID, 15.6 microg per disc). All the extracts showed strong antioxidant activity comparable with or higher that of alpha-tocopherol.

Chemopreventive effects of coumaperine from pepper on the initiation stage of chemical hepatocarcinogenesis in the rat.

This study was designed to investigate the chemopreventive action of three natural products, coumaperine, aurapten and an extract from rosemary, against the initiation stage of rat hepato-carcinogenesis. Coumaperine has been isolated from white pepper as a naturally occurring antioxidative agent, but its potential modifying effects on carcinogenesis remain unclear. In experiment 1, a modification of the model developed by Tsuda et al. was applied, with assessment of numbers and areas of induced glutathione S-transferase placental form (GST-P)-positive hepatocellular foci in male F344 rats. Coumaperine, aurapten and the extract from rosemary were administered i.g. at 100 mg / kg / day once daily for 5 days with initiation by diethylnitrosamine (DEN) on day 4 (20 mg / kg, i.p.). Numbers and areas of GST-P-positive foci in each group given test chemicals tended to be decreased as compared to the vehicle control group values, significance being achieved for number with coumaperine. Experiment 2 was planned to investigate the mechanism of the inhibitory effects of coumaperine. Livers at 8 h after initiation by DEN were examined with coumaperine administered at 100 mg / kg / day once daily for 3 days. Proliferating cell nuclear antigen (PCNA)-positive cells tended to be decreased as compared to the vehicle control, but no effects on apoptosis or cytochrome P-450 (CYP) 2E1 expression were apparent. Our results suggest that coumaperine provides protection against initiation of hepatocarcinogenesis, and that this is related to inhibition of cell proliferation.