Central PACAP mediates the sympathetic effects of leptin in a tissue-specific manner.

We previously demonstrated that the peptidergic neurotransmitter pituitary adenylate cyclase-activating polypeptide (PACAP) affects the autonomic system and contributes to the control of metabolic and cardiovascular functions. Previous studies have demonstrated the importance of centrally-mediated sympathetic effects of leptin for obesity-related hypertension. Here we tested whether PACAP signaling in the brain is implicated in leptin-induced sympathetic excitation and appetite suppression. In anesthetized mice, intracerebroventricular (ICV) pre-treatment with PACAP6-38, an antagonist of the PACAP receptors (PAC1-R and VPAC2), inhibited the increase in white adipose tissue sympathetic nerve activity (WAT-SNA) produced by ICV leptin (2µg). In contrast, leptin-induced stimulation of renal sympathetic nerve activity (RSNA) was not affected by ICV pre-treatment with PACAP6-38. Moreover, in PACAP-deficient (Adcyap1-/-) mice, ICV leptin-induced WAT-SNA increase was impaired, whereas RSNA response was preserved. The reductions in food intake and body weight evoked by ICV leptin were attenuated in Adcyap1-/- mice. Our data suggest that hypothalamic PACAP signaling plays a key role in the control by leptin of feeding behavior and lipocatabolic sympathetic outflow, but spares the renal sympathetic traffic.

The effects of microbial materials adhered to Asian sand dust on allergic lung inflammation.

Asian sand dust (ASD) containing microbiological materials, sulfate (SO(4)(2)), and nitrate (NO(3)(-) ) derived from air pollutants in East China, reportedly cause adverse respiratory health effects. ASD aggravates ovalbumin (OVA)-associated experimental lung eosinophilia. In this study, the toxic materials adsorbed onto ASD were excluded by heat treatment at 360 degrees C for 30 min. The effects of nonheated ASD or heated ASD (H-ASD) toward the allergic lung inflammation were compared in murine lungs. ICR mice were administered intratracheally with normal saline (control), H-ASD, ASD, OVA, OVA + H-ASD, and OVA + ASD, four times at 2-week intervals. ASD only increased neutrophils in bronchoalveolar lavage fluids (BALFs) along with pro-inflammatory mediators, such as keratinocyte chemoattractant (KC). H-ASD and ASD enhanced eosinophil recruitment induced by OVA in the alveoli and in the submucosa of the airway, which has a goblet cell proliferation in the bronchial epithelium. The two ASDs synergistically increased interleukin-5 (IL-5), monocyte chemotactic protein-3 (MCP-3), and eotaxin, which were associated with OVA, in BALF. The enhancing effects were much greater in ASD than in H-ASD. The two ASDs induced the adjuvant effects to specific IgE and IgG1 production by OVA. In the in vitro study using RAW264.7 cells, ASD increased the expression of Toll-like receptor 2 (TLR 2) mRNA but not TLR4 mRNA. H-ASD caused no expression of either TLR mRNA. These results suggest that the aggravated lung eosinophilia by ASD may be due to activation of Th2-associated immune response via the activation of TLR2 by microbial components adhered to ASD.

Murine strain differences in 8-hydroxy-deoxyguanosine formation in hepatic DNA induced by oxidized lard and dietary oils.

Difference of 8-hydroxy-deoxyguanosine (8-OH-dG) formation in liver DNA in C3H/HeN and in C57BL/6 mice--fed oxidized lard and dietary oils (soybean and sardine)--was investigated. The blank levels of 8-OH-dG were higher in C3H/HeN mice (highly sensitive to liver tumorigenesis) than in C57BL/6 mice (resistant strain). The level of 8-OH-dG increased much more in C3H/HeN mice than in the C57BL/6 mice fed by oxidized lard and dietary oil treatment. Feeding oxidized lard and dietary oils increased 8-oxo-guanine DNA glycosylase I (OGG1) and mRNA 8-oxo-dGTPase in C57BL/6 mice. On the other hand, no appreciable change of mRNA in the C3H/HeN mice was observed. The formation differences of 8-OH-dG from the two murine strains fed with oxidized lard and dietary oils may be associated with the different mRNA levels in the DNA repair enzymes because the mRNA levels in the DNA repair enzymes were much lower in C3H/HeN mice than in C57BL/6 mice.

Liver carcinogenesis and formation of 8-hydroxy-deoxyguanosine in C3H/HeN mice by oxidized dietary oils containing carcinogenic dicarbonyl compounds.

Oxidized dietary oils (lard, soybean oil, and sardine oil) were orally administered to C3H/HeN male mice. After 6 months, benign hepatocellular adenoma was observed in the mice treated with all three oxidized dietary oils. After 12 months, malignant hepatocellular carcinoma and hepatoblastoma were observed in addition to the benign tumor. Oxidized sardine oil caused the highest tumor incidence (35%) and malignant tumors (27.5%) among the oxidized dietary oils tested. Mice treated with oxidized lard and sardine oil exhibited a significant increase of 8-OH-dG in the livers. The amounts of 8-OH-dG found in the mice treated with oxidized sardine oil correlated with the rates of tumor incidence. After 6 months, mRNA decreased in the case of oxidized lard and sardine oil, whereas it increased in the case of oxidized soybean oil, either in 8-oxoguanine-DNA glycosylase (OGG1) or in 8-oxo-dGTPase. On the other hand, there was no appreciable change in mRNA, in either OGG1 or 8-oxo-dGTPase, after 12 months. Oxidized sardine oil contained the highest level of malonaldehyde (MA) (713+/-91.1 nmol/g) and glyoxal (33.3+/-5.2 nmol/g) among three oxidized oils. The malignant tumor incidence correlated with the high level of MA and glyoxal found in the dietary oils tested.

Inhibition of malonaldehyde formation from blood plasma oxidation by aroma extracts and aroma components isolated from clove and eucalyptus.

The inhibitory effect of aroma extracts isolated from clove buds [Syzygium aromaticum (L.) Merr. et Perry] and eucalyptus leaves (Eucalyptus polyanthemos Schauer) on malonaldehyde (MA) formation from horse blood plasma oxidized with Fenton's reagent was determined by gas chromatography. Aroma chemicals such as eugenol, thymol and benzyl alcohol, identified in the aroma extracts, were examined for their inhibitory effect on the same system. Between the two aroma extracts tested, clove exhibited the most potent antioxidant activities. Extracts of eucalyptus and clove inhibited MA formation by 23 and 48%, respectively, at the level of 400 microg/ml, whereas, alpha-tocopherol and BHT inhibited MA formation by 52 and 70%, respectively, at the same level. Eugenol, thymol and benzyl alcohol inhibited MA formation by 57, 43 and 32%, respectively, at the level of 400 microg/ml.

Antioxidative activity of volatile chemicals extracted from beer.

Volatile chemicals obtained from a commercial beer by liquid-liquid continuous extraction were evaluated for antioxidant activity. The inhibitory ability of this extract toward the conversion of hexanal to hexanoic acid was monitored over a 35-day period. The volatile extract demonstrated >99% effectiveness at inhibiting hexanal oxidation at 50 microg/mL, comparable to that of the natural antioxidant alpha-tocopherol (vitamin E). Volatile compounds contained in the extract were isolated and identified by gas chromatography-mass spectrometry (GC-MS). From the volatile constituents identified in beer extract, phenylethyl alcohol, maltol, and 2-furanmethanol were examined for antioxidative activities. At a concentration of 500 microg/mL, maltol and 2-furanmethanol demonstrated approximately 95 and 100% inhibition of hexanal oxidation over 35 days, respectively. Phenylethyl alcohol did not show any appreciable level of inhibition of hexanal oxidation. Heterocyclic compounds, some of which are known to possess antioxidative activities, were also identified in the volatile extract.

Variation of major volatile constituents in various green teas from Southeast Asia.

A total of 15 green tea samples were prepared from fresh tea leaves obtained from three different countries: two from Laos, seven from Myanmar, and six from Vietnam. The volatile aroma constituents of the 15 samples were analyzed by gas chromatography/mass spectroscopy. Eleven aroma constituents were chosen from over 100 chemicals found in the samples to compare differences among various teas. They were hexanal, 1-penten-3-ol, heptanal, 1-pentenal, (Z)-2-penten-1-ol, (Z)-3-penten-1-ol, linalool oxide (trans-furanoid), linalool oxide (cis-furanoid), linalool, linalyl propanoate, and geraniol. Generally, concentrations of linalool and hexanal seem to play an important role in the quality of green teas. Green teas from Laos and Myanmar contained heterocyclic compounds, such as pyridines and pyrazines, formed by high-temperature processing. The presence of these heterocyclic compounds suggested that the temperature used for tea processing plays an important role in the formation of aroma chemicals in green teas.

Antioxidative activities of heterocyclic compounds formed in brewed coffee.

Typical volatile heterocyclic compounds found in brewed coffee extracts-pyrroles, furans, thiophenes, and thiazoles-were examined for antioxidative activity, which was determined by measuring the oxidative conversion of hexanal to hexanoic acid using gas chromatography. 2-Acetylpyrrole, 1-methylpyrrole, and pyrrole inhibited hexanal oxidation by 98, 87, and 78%, respectively, at a concentration of 500 microgram/mL over a period of 30 days. 2-Methylfuran, which inhibited hexanal oxidation by 90% at all concentrations tested (500, 200, and 100 microgram/mL) for a 30-day period, exhibited the greatest activity among furans tested. Similarly, 2-methylthiophene, which inhibited hexanal oxidation by almost 100% at a concentration of 500 microgram/mL over 30 days, exhibited the greatest activity among the thiophenes tested. In general, thiazoles were ineffective antioxidants at all concentrations tested. However, 4,5-dimethylthiazole was able to inhibit hexanal oxidation by 50% at the highest level tested (500 microgram/mL). 2-Acetylpyrrole, 2-methylfuran, and 2-methylthiophene at concentrations of 500, 200, and 100 microgram/mL and furan at a concentration of 500 microgram/mL exhibited antioxidative activities comparable to that of the synthetic antioxidant butylated hydroxytoluene at a concentration of 50 microgram/mL.

Determination of antioxidant properties of aroma extracts from various beans.

Aroma extracts from fresh soybeans, mung beans, kidney beans, and azuki beans were prepared using simultaneous steam distillation and solvent extraction (SDE) under mild conditions (55 degrees C and 95 mmHg). Extracts were examined for antioxidative activities in two different assays. The aroma extracts isolated from all beans inhibited the oxidation of hexanal for nearly one month at a level of 250 microL/mL. Mung bean and soybean extracts inhibited malonaldehyde (MA) formation from cod-liver oil by 86% and 88%, respectively, at the 250 microL/mL level. Azuki and kidney bean extracts inhibited MA formation from cod-liver oil by 76% and 53%, respectively, at the 250 microL/mL level. The antioxidative activities of mung bean and soybean extracts were comparable with that of the natural antioxidant, alpha-tocopherol (vitamin E).

Antioxidant properties of aroma compounds isolated from soybeans and mung beans.

Aroma compounds contained in the extracts of soybean and mung bean that possess antioxidant activity were identified by gas chromatography (GC) and gas chromatography/mass spectrometry (GC/MS). The major aroma constituents of soybeans were 1-octen-3-ol (13.699 ppm), maltol (1.662 ppm), phenylethyl alcohol (1.474 ppm), hexanol (1.430 ppm), and gamma-butyrolactone (1.370 ppm). The major aroma constituents of mung beans were hexanol (3.234 ppm), benzyl alcohol (2.060 ppm), gamma-butyrolactone (1.857 ppm), 2-methyl-2-propenal (1. 633 ppm), and pentanol (1.363 ppm). The major aroma chemicals of soybeans and mung beans were examined for antioxidative activities in two different assays. Eugenol, maltol, benzyl alcohol, and 1-octen-3-ol showed potent antioxidative activities in two different assays. Eugenol, maltol, benzyl alcohol, and 1-octen-3-ol inhibited the oxidation of hexanal by 100%, 93%, 84%, and 32%, respectively, for a period of 40 days at the 500 microg/mL level. Eugenol, maltol, benzyl alcohol, and 1-octen-3-ol inhibited malonaldehyde (MA) formation from cod liver oil by 91%, 78%, 78%, and 78%, respectively, at the 160 microg/mL level. The antioxidative activity of eugenol was comparable to that of the natural antioxidant alpha-tocopherol (vitamin E).

Volatile chemicals identified in extracts from leaves of Japanese mugwort (Artemisia princeps pamp.).

Extracts from leaves of Japanese mugwort (Artemisia princeps Pamp.) were obtained using two methods: steam distillation under reduced pressure followed by dichloromethane extraction (DRP) and simultaneous purging and extraction (SPSE). A total of 192 volatile chemicals were identified in the extracts obtained by both methods using gas chromatography (GC) and gas chromatography-mass spectrometry (GC-MS). They included 47 monoterpenoids (oxygenated monoterpenes), 26 aromatic compounds, 19 aliphatic esters, 18 aliphatic alcohols, 17 monoterpenes (hydrocarbon monoterpenes), 17 sesquiterpenes (hydrocarbon sesquiterpenes), 13 sesquiterpenoids (oxygenated sesquiterpenes), 12 aliphatic aldehydes, 8 aliphatic hydrocarbons, 7 aliphatic ketones, and 9 miscellaneous compounds. The major volatile constituents of the extract by DRP were borneol (10.27 ppm), alpha-thujone (3.49 ppm), artemisia alcohol (2.17 ppm), verbenone (1.85 ppm), yomogi alcohol (1.50 ppm), and germacren-4-ol (1.43 ppm). The major volatile constituents of the extract by SPSE were 1,8-cineole (8.12 ppm), artemisia acetate (4.22 ppm), alpha-thujone (3.20 ppm), beta-caryophyllene (2.39 ppm), bornyl acetate (2.05 ppm), borneol (1.80 ppm), and trans-beta-farnesene (1. 78 ppm).

Antioxidative activities of aroma extracts isolated from natural plants.

Natural leaves and flowers containing numerous aroma chemicals are widely used in aromatherapy since ancient times. In addition to their pleasant smells, aroma chemicals might have some beneficial health effects. Aroma extracts, isolated from coffee beans, soybeans, and mung beans by steam distillation under mild conditions (55 degrees C and 85 mm Hg) were examined for their antioxidative activities. The inhibitory effect of these extracts toward hexanal/hexanoic acid conversion was measured in the testing solution over prolonged time periods. The inhibitory effects of these extracts toward malonaldehyde formation from lipids oxidized by Fenton's reagent were also measured. The antioxidative activity of these extracts, in particular coffee bean extract, was consistent with that of BHT or alpha-tocopherol (vitamin E). Soybeans and mung beans extract contained maltol, which inhibits hexanal oxidation significantly. Eugenol, which is one of the major constituents of mung bean extract, exhibited potent antioxidative activity in an aldehyde/carboxylic acid assay. Antioxidants such as eugenol and maltol may play an important role in the pharmaceutical activities of natural plant extracts used for aromatherapy.

Aroma chemicals isolated and identified from leaves of Aloe arborescens Mill. Var. Natalensis Berger.

Extracts from leaves of aloe (Aloe arborescens Mill. var. natalensis Berger) were obtained using two methods: steam distillation under reduced pressure followed by dichloromethane extraction (DRP) and simultaneous purging and extraction (SPE). A total of 123 aroma chemicals were identified in the extracts obtained by both methods using gas chromatography and gas chromatography/mass spectrometry. There were 42 alcohols, 23 terpenoids, 21 aldehydes, 9 esters, 8 ketones, 6 acids, 5 phenols, and 9 miscellaneous compounds. The major aroma constituents of this extract by DRP were (Z)-3-hexenol (29.89%), (Z)-3-hexenal (18.86%), (E)-hexenal (7.31%), 4-methyl-3-pentenol (5.66%), and butanol (4.29%). The major aroma constituents of this extract by SPE were (E)-2-hexenal (45.46%), (Z)-3-hexenal (32.12%), hexanal (9.14%), (Z)-3-hexenol (1.60%), and 3-pentanone (1.41%). Terpenoids were also found as one of the major constituents. The fresh green note of aloe leaves is due to the presence of these C(6) alcohols and aldehydes as well as terpenoids.

Lecithinized superoxide dismutase attenuates phorbol myristate acetate-induced injury in isolated dog lung.

Lecithinized superoxide dismutase, a lecithin derivative bound to recombinant human CuZn superoxide dismutase, has a higher affinity for cells such as polymorphonuclear leukocytes and endothelial cells than recombinant human CuZn superoxide dismutase has. We determined the protective effects of lecithinized superoxide dismutase on the increased microvascular permeability induced by phorbol myristate acetate (PMA) in isolated dog lungs. Microvascular permeability was assessed by the capillary filtration coefficient (Kf,c) and solvent drag reflection coefficient (sigma(f)). PMA (13.3 microg) increased microvascular permeability, as evidenced by an increase in Kf,c and the small sigma(f) value. Lecithinized superoxide dismutase at both low (4800 U) and high doses (48,000 U) inhibited the PMA-induced increase in Kf,c, but only the high dose of lecithinized superoxide dismutase attenuated the decrease in sigma(f). Recombinant human CuZn superoxide dismutase did not affect the PMA-induced increase in vascular permeability at either a low (4800 U) or a high dose (48,000 U). These findings suggest that lecithinized superoxide dismutase has a protective effect against oxygen radical-induced lung injury in isolated dog lungs.

Simultaneous determination of acrolein, malonaldehyde and 4-hydroxy-2-nonenal produced from lipids oxidized with Fenton's reagent.

Ethyl linoleate, ethyl linolenate, ethyl arachidonate and cod liver oil were oxidized with Fenton's reagent. Acrolein, malonaldehyde and 4-hydroxynonenal formed were derivatized to N-methylpyrazoline, N-methylpyrazole and 5-(1'-hydroxyhexyl)-1-methyl-2-pyrazoline with N-methylhydrazine, respectively. The derivatives were simultaneously analysed by gas chromatograph equipped with a fused silica capillary column and a nitrogen-phosphorus detector. The maximum amounts of acrolein (9.7 +/- 2.11 nmol/ml) and malonaldehyde (61.18 +/- 6.51 nmol/ml) were formed from cod liver oil. The highest amount of 4-hydroxynonenal (6.83 +/- 0.53 nmol/ml) was produced from ethyl arathidonate.

Effects of Cassia obtusifolia (sicklepod) extracts and anthraquinones on muscle mitochondrial function.

Naturally occurring quinones and quinone-containing extracts of seeds of the toxic plant Cassia obtusifolia (sicklepod) affected muscle mitochondrial function. Aqueous suspensions and organic extracts of C. obtusifolia seeds slightly elevated plasma creatine kinase levels of Sprague-Dawley rats. These extracts were analyzed by fused silica capillary gas chromatography and found to contain nine anthraquinones and three anthrones. Urinary metabolites primarily consisted of beta-glucuronide conjugates of the anthraquinones. The three anthrones or conjugate analogues were not present in the urine in detectable amounts. Emodin, doxorubicin and organic extracts of C. obtusifolia inhibited NADH:cytochrome c oxidoreductase activity of bovine heart mitochondrial particles and NADH:CoQ oxidoreductase activity of porcine heart mitochondrial NADH dehydrogenase, whereas juglone was stimulatory. Relative quinone metabolism correlated with semiquinone formation rate and with redox potential. A protective effect of coenzyme Q against enzyme inhibition by anthraquinones was also observed.

Genotoxicity testing of Maillard reaction products.

Since the development of short-term genotoxicity tests such as the Ames assay, the mutagenicity of Maillard reaction products has been tested extensively. Some products have exhibited strong activity. For example, one of the earliest studies demonstrated some mutagenic activity in a dichloromethane extract of a D-glucose/ammonia Maillard model system. Many researchers have attempted to pinpoint the principal chemical(s) of mutagenicity of the Maillard products using various sugar-amino acid browning model systems over last two decades. However, no mutagenic individual Maillard product has been isolated and identified. Nitrite has been also used as a reactant in browning reaction model systems, primarily to investigate the formation of potentially mutagenic or carcinogenic N-nitroso compounds. Recently some potent mutagens isolated from pyrolyzed amino acids or proteins have begun to receive attention as Maillard reaction products.

Analysis of free malondialdehyde in photoirradiated corn oil and beef fat via a pyrazole derivative.

Malondialdehyde (MA) formed in linolenic acid, linoleic acid, corn oil and beef fat upon photoirradiation was determined by gas chromatography (GC). The MA produced was reacted with methylhydrazine to give 1-methylpyrazole and was subsequently analyzed on a GC equipped with a nitrogen-phosphorus specific detector and a fused silica capillary column. MA values determined by this method correspond to free or unbound MA levels. Linolenic and linoleic acids produced 867 micrograms MA/g and 106 micrograms MA/g, respectively. Oleic and stearic acids did not produce detectable levels of MA upon photoirradiation. Amounts of MA produced after eight hour irradiations of corn oil and beef fat were 56.24 micrograms/g and 25.01 micrograms/g, respectively. Some photoreaction products in irradiated corn oil also were identified as methylhydrazine derivatives.

Biological and chemical studies on overheated brewed coffee.

Vapour formed from overheated decaffeinated coffee was condensed and tested for mutagenicity using the Ames assay in Salmonella typhimurium strains TA98 and TA100. Vapour produced at 73 and 100 degrees C exhibited no mutagenicity. The basic fraction of vapour produced at 350 degrees C showed weak mutagenicity towards strains TA98 with metabolic activation. The chemical analysis of this fraction identified pyridines and pyrazines as the major constituents. None of the compounds identified in this fraction has been reported as mutagenic when tested in the Ames assay.