We previously found that malathion residue in wheat kernels was decomposed by wheat kernel carboxylesterase (CE) during sample preparation for pesticide residue analysis. The degradation of malathion in supernatant of wheat kernel homogenate was enzyme-kinetically analyzed to characterize the CE. Malathion α-monocarboxylic acid (MMCα), malathion β-monocarboxylic acid (MMCβ), malathion dicarboxylic acid (MDC) and desmethyl malathion (DMal) were identified by analysis using liquid chromatography/electrospray ionization mass spectrometry (LC/ESI-MS) after incubation at 36°C of the reaction mixture of malathion in the supernatant of wheat kernel homogenate. Since small amounts of DMal and MMCβ were formed in boiled supernatant of wheat kernel homogenate, these compounds were nonenzymatically produced. Examination of the time course of enzymatic malathion degradation indicated that malathion was decomposed into MDC through both MMCα and MMCβ, and the amount of MMCα formed was greater than that of MMCβ at maximum concentrations of both MMC isomers. MMCα and MMCβ were enzymatically decomposed into only MDC, which was the final metabolite of malathion because there was no further decomposition product. The order of degradation half-lives by wheat kernel CE (malathion<MMCβ<MMCα) is consistent with the intensity of their hydrophobicity. These results suggest that when malathion is degraded via MMC to MDC by wheat kernel CE, its enzymatic degradability and the decomposition products depends on their hydrophobicity.
Titanium dioxide (TiO2)-assisted photodegradation of volatile organic compounds (VOCs), especially malodorous substances at ppb level in closed air are reported. As target VOCs we selected acetaldehyde, dimethyl disulfide (DMDS), dimethyl sulfide (DMS) and methyl mercaptan, because of their toxicity and unpleasant odor at low concentration in closed air. The initial concentrations were fixed at 0.4-5.0 ppm for acetaldehyde, 0.03 ppm for DMDS and DMS and 0.02 ppm for methyl mercaptan respectively. A blacklight UV-lamp was employed as a light source and the intensity of UV-light was controlled at 1.0 mW/cm2 (351 nm). The half life of these odorous substances was from 20 min to 120 min and pseudo-first-order reaction rates were 1.1×10-2 min-1 (acetaldehyde), 1.4×10-2 min-1 (DMDS), 4.8×10-2 min-1 (DMS) and 8.0×10-3 min-1 (methyl mercaptan). One of the by-products of TiO2 photodegradation of acetaldehyde was identified as formaldehyde by liquid chromatography/mass spectrometry (LC/MS). These basic data of TiO2 photodegradation for odorous substances will be useful for the construction of an air purification system using TiO2 photodegradation in closed air, for example in living rooms, hospitals and care rooms.
Pantoprazole is an excellent proton pump inhibitor for the treatment of acid-related lesions. In this study we compared the pharmacodynamic effects of (±)-pantoprazole sodium [(±)-PAN·Na] and its enantiomers on gastric acid secretion and their possible side effects on gastric mucus, mucosa, and gastric endocrine cells. (±)-PAN·Na, (-)-PAN·Na, and (+)-PAN·Na dose-dependently inhibited the secretion of basal gastric acid and histamine-induced gastric acid, and (-)-PAN·Na showed the most potent effect, which was confirmed in an in vivo experiment in rabbits and in an in vitro experiment using the stomachs of juvenile rats. On the other hand, (-)-PAN·Na, (+)-PAN·Na, and (±)-PAN·Na did not influence significantly the free and barrier mucus content in rats with short-term administration for 3 days. No obvious differences were observed in serum gastrin levels, volume densities of parietal cells, G cells and D cells, mucosal thickness, or relative stomach weight and body weight among the rats administered (-)-PAN·Na, (+)-PAN·Na, or (±)-PAN·Na for 40 days. Direct administration of (-)-PAN·Na can inhibit gastric acid secretion more effectively with no increase in side effects.
The alteration of the GABAergic system in dimethoate intoxication was explored. A rat brain AChE (acetyl-cholinesterase) activity and GABAergic profiles, including the level of gamma-aminobutyric acid (GABA) and the densities and affinities of gamma-aminobutyric acid A (GABAA) receptors in the hippocampus, were examined 2 hr after dimethoate administration (0, 38.9, 83.7 and 180.0 mg/kg bw po). The AChE activity was reduced by 60-70% by all three doses of dimethoate. Statistically significant elevation of GABA levels was observed following the administration of dimethoate 180.0 mg/kg bw (121.5% of control, p<0.05), while the Bmax values for the GABAA receptors of the hippocampal synaptic membranes were 50.6% and 51.6% of control values in the 83.7 and 180.0 mg/kg bw dimethoate groups, respectively (p<0.05). Statistically significant decreases in the Kd values of 19.8, 51.7, and 53.4% vs. controls were observed in a dose-dependent manner (p<0.05). In conclusion, it is suggested that the GABAergic system is maybe involved in the neurotoxicity of organophosphates as well as the cholinergic mechanisms.
Although many physiological changes after space flight have been reported, it is not clear how microgravity influences our bodies. The focus of the present study was to clarify the changes in G-protein-coupled receptor-mediated intracellular signaling, especially Gs-adenylyl cyclase (AC)-adenosine 3', 5'-cyclic monophosphate (cyclic AMP) pathway, under simulated microgravity. Human astrocytoma 1321N1 cells were cultivated under vector-averaged microgravity conditions generated by clinostat rotation (20 rpm) for 24 hr. Isoproterenol, a β-adrenergic agonist and forskolin, a direct AC stimulant, increased intracellular cyclic AMP level in concentration dependent manners, however, both of which response were decreased in cells cultivated in clinostat rotation. While the level of Gαs or intracellular ATP, a substrate for AC, was not changed, the AC activity was significantly low in the membranes of clinostat-rotated cells. Reverse transcriptase-polymerase chain reaction (RT-PCR) analysis revealed that AC type 3 (AC3), AC6, and AC9 and to a lesser extent AC7 and AC8 were expressed in 1321N1 cells. Among them, the expression of AC6 mRNA was significantly decreased by clinostat rotation. These results indicate that intracellular cyclic AMP production by agonists may be decreased via a reduction in AC6 expression under simulated microgravity conditions.
To elucidate a physiological function of dried-bonito broth (DBB) on blood fluidity and oxidative stress, we performed a randomized double-blind placebo-controlled study in twenty-four healthy adult subjects. The subjects ingested DBB or a placebo for four weeks, and blood fluidity and oxidative stress were measured before and after ingestion. Blood fluidity was measured using a microchannel array flow analyzer by the passage time of 100 μl of heparinized whole blood through the microchannel array, while oxidative stress was evaluated as a level of derivative of reactive oxygen metabolites (d-ROMs) by a free radical analysis system (FRAS). DBB ingestion significantly shortened the blood passage time from 55.4±3.4 to 47.6±2.0 sec (mean±SEM, p<0.05), while no significant change was observed in the placebo group (52.4±3.4 to 51.4±2.6 sec, mean±SEM) indicating that DBB ameliorated blood fluidity. The level of d-ROMs, known as a biomarker of oxidative stress, significantly decreased after DBB ingestion from 337.2±18.5 to 316.5±12.9 Carrotelli units (Carr. U.) (mean±SEM, p<0.05), suggesting that DBB reduced oxidative stress. Among subjects with a d-ROMs score >320, regarded as being in a state of oxidative stress, changes in blood fluidity tended to correlate with changes in d-ROMs score (ρ=0.55, p=0.06), showing that blood fluidity may have improved in subjects whose oxidative stress was markedly decreased. These results also showed a possibility that DBB ingestion improved blood fluidity by decreasing oxidative stress. In previous studies, daily DBB ingestion improved various fatigue-related symptoms, so we investigated the effect of DBB on fatigue-related symptoms via a questionnaire survey in the present study. The result of this survey showed that symptoms of shoulder stiffness and visual fatigue were improved only in the DBB group (p<0.05, p<0.1, respectively). Insufficient blood circulation is considered to lead to the development of shoulder stiffness, visual fatigue, and other fatigue-related symptoms. Based on these findings, we considered that dietary intake of DBB may improve blood fluidity by reducing oxidative stress and thus might protect against fatigue.
Nonylphenol, which is used industrially as a surfactant, is an endocrine-disrupting chemical (EDC) which has estrogenic activity. The novel biotransformation of nonylphenol was investigated, based on our previously reported ipso-metabolism of para-substituted phenols by cytochrome P450 (P450). Three novel metabolites of nonylphenol, i.e., nonylquinol, 4'-hydroxynonanophenone (CO-NP) as benzyl-oxidized nonylphenol, and hydroquinone, were detected in a rat liver microsome reaction mixture. On the other hand, production of 1-(4'-hydroxyphenyl)nonan-1-ol (OH-NP), namely benzyl-hydroxylated nonylphenol, was detected in a human liver microsome reaction mixture. The formation of all these metabolites was suppressed by the addition of P450 inhibitor. This showed that all nonylphenol metabolism was catalyzed by P450. To identify which P450 isoenzyme is involved in each reaction, fourteen human P450 (CYP) isozymes, CYP1A1, 1A2, 2A6, 2B6, 2C8, 2C9, 2C18, 2C19, 2D6, 2E1, 3A4, 3A5, 3A7, and CYP4A11, were examined. CYP1A1, 1A2, and CYP2B6 effectively catalyzed the production of nonylquinol. CYP2B6 also catalyzed the benzyl-hydroxylation to give OH-NP. Hydroquinone was formed mainly from OH-NP, not via CO-NP. We examined the estrogenic activity of these new metabolites by estrogen receptor (ER)-binding reporter gene assay. Nonylquinol, OH-NP and hydroquinone have no ER-binding activity. However, CO-NP showed the same level of estrogen receptor binding activity as nonylphenol. Moreover, the amount of CO-NP formed was small. Therefore, the novel metabolic pathways led overall to metabolic inactivation, as concerns the estrogenic activity of nonylphenol through the ER.
Estrogenic/antiestrogenic activities of 14 polycyclic aromatic hydrocarbons (PAHs) and 63 monohydroxylated PAHs (OHPAHs) having 2 to 6 rings were evaluated by yeast two-hybrid assay expressing human estrogen receptor α. Relative effective potencies of estrogenic and antiestrogenic activities were calculated as the inverse values of the relative concentration of the test compound that gave the same activities of E2 and 4-hydroxytamoxifen, respectively. PAHs did not show any estrogenic/antiestrogenic activity, but several OHPAHs having 3 to 5 rings showed activities. Especially, OHPAHs having 4 rings such as 3-, 4- and 10-hydroxybenz[a]anthracenes (3-, 4- and 10-OHBaAs) and 2-hydroxychrysene (2-OHCh) showed strongly estrogenic activity. Several other OHPAHs having 4 rings such as 2- and 3-hydroxybenzo[c]phenanthrenes (2-, 3- OHBcPhs), 2-OHBaA and 3-OHCh showed strongly antiestrogenic activity. The length-to-breadth (L/B) ratios of the rectangular van der Walls planes surrounding the ring molecules of estrogenic OHPAHs were in the narrow range from 1.599 to 1.734. The distances between the oxygen atom of the phenol group and farthest hydrogen atom (O-H distance) of the estrogenic OHPAHs ranged from 10.825 Å to 11.738 Å. The L/B ratios and O-H distances of antiestrogenic OHPAHs were in the wider ranges from 1.277 to 1.734 and from 8.47 Å to 11.681 Å, respectively. The partial charges (atomic unit) of the phenol group of both estrogenic and antiestrogenic OHPAHs were in the range from -0.250 atmic unit (au) to -0.253 au. The similarity of these values to those of E2 and diethylstilbestrol suggested that the compositions of estrogenic OHPAHs were similar to them and that the compositional conditions of estrogenic OHPAHs were much smaller than those of antiestrogenic OHPAHs. These results raise the possibility of predicting the estrogenic/antiestrogenic activities of OHPAHs from their structural characteristics, although using only the above three parameters might not be enough for accurate estimations.
The effect of bee pollen Cistus ladaniferus extract on ovariectomy (OVX)-induced bone loss in vivo was investigated. The water-solubilized extracts were obtained from the bee pollen of Cistus ladaniferus. Cistus extract (5.0 or 10.0 mg/100 g body weight) was orally administered once daily for 30 days to OVX rats. The analysis using a peripheral quantitative computed tomography (pQCT) showed that OVX-induced a significant decrease in mineral content, mineral density, and polar strength strain index in the femoral-metaphyseal tissues. These decreases were significantly prevented after the administration of cistus extract (10.0 mg/100 g). Moreover, OVX-induced a significant decrease in calcium content in the femoral-diaphyseal and -metaphyseal tissues. This decrease was significantly prevented after the administration of cistus extract (5.0 or 10.0 mg/100 g). This study demonstrates that cistus extract has a preventive effect on OVX-induced bone loss in vivo.
Ferrous ferric chloride (FFC) is a special aqueous iron which is a complex of ferrous chloride and ferric chloride and has a function in both oxidation and reduction. FFC is thought to stimulate the cellular function of living organisms. Although FFC is reported to stimulate the function of red blood cells, it has not been determined whether FFC stimulates the function of skin cells. To understand the role of FFC in the function of skin cells, we studied its effects on the proliferation and differentiation of keratinocytes and melanoblasts or melanocytes. FFC was added to a serum-free culture of neonatal mouse epidermal cells and its effects on the proliferation and differentiation of keratinocytes and melanoblasts or melanocytes were investigated. FFC stimulated the proliferation and differentiation of keratinocytes and melanoblasts or melanocytes. The proliferation of keratinocytes and that of melanoblasts or melanocytes was stimulated to the same extent (a two-fold increase), suggesting that the proliferation of the two types of cells constituting the epidermis may be equally stimulated by FFC. These results suggest that FFC may activate skin function by promoting cell renewal via the stimulation of the proliferation and differentiation of keratinocytes and melanoblasts or melanocytes.
5-Methoxy-N,N-diisopropyltryptamine (5-MeO-DIPT), a psychotomimetic tryptamine derivative, and its relevant metabolites have been determined in eleven urine specimens from six 5-MeO-DIPT users, and their excretion profiles have been investigated by gas chromatography/mass spectrometry (GC/MS) and liquid chromatography/mass spectrometry (LC/MS). Three metabolites, 5-hydroxy-N,N-diisopropyltryptamine (5-OH-DIPT), 6-hydroxy-5-methoxy-N,N-diisopropyltryptamine (6-OH-5-MeO-DIPT), and 5-methoxy-N-isopropyltryptamine (5-MeO-NIPT) were determined in the urine specimens. Urinary conjugated metabolites, both sulfates and glucuronides of 5-OH-DIPT and 6-OH-5-MeO-DIPT, were hydrolyzed completely by the use of Helix pomatia sulfatase/β-glucuronidase. Degradation of 6-OH-5-MeO-DIPT during incubation for hydrolysis was successfully prevented by the addition of ascorbic acid. The hydrolysis treatment increased the detection amounts of 5-OH-DIPT and 6-OH-5-MeO-DIPT in most of the specimens, and the increase in 6-OH-5-MeO-DIPT was more drastic than that in 5-OH-DIPT. The concentrations of 5-MeO-DIPT (<1.7 μg/ml) and 5-MeO-NIPT (<3.5 μg/ml) were lower than those of 5-OH-DIPT (0.01-47 μg/ml) and 6-OH-5-MeO-DIPT (<69 μg/ml) detected after hydrolysis (the totals of their free and conjugated forms). These metabolites were detectable over longer periods post intake than the parent drug; 35 hr for 5-MeO-DIPT, 80 hr for 5-OH-DIPT, and 60 hr for 6-OH-5-MeO-DIPT and 5-MeO-NIPT.
Catechins are naturally occurring polyphenols, which are supposed to have antioxidative effects on foods and living cells. We examined the antioxidative effects of (+)-catechin and 3-O-acyl derivatives of (+)-catechin in combination with copper(II) [Cu(II)] ions on the formation of carbonyl groups in bovine serum albumin (BSA) at pH 7.4 by incubating in vitro for 90 min at 37°C. In the presence of 2,2'-azobis(2-amidinopropane) dihydrochloride (AAPH), a free radical generator, the formation of carbonyl groups in BSA, which increased time-dependently, was significantly (p<0.05) attenuated by the addition of (+)-catechin and 3-O-acyl-(+)-catechins. However, Cu(II) ions, but not Zn(II) ions, blocked the antioxidative effects of (+)-catechin and 3-O-octanoyl-(+)-catechin on the oxidative modification of BSA induced by AAPH. On the other hand, Cu(II) ions had no influence on the scavenging activity of (+)-catechin for 2,2'-azinobis(3-ethylbenzothiazoline-6-sulphonate) cation radicals. Furthermore, in the absence of AAPH, coexistence of Cu(II) ions with (+)-catechin greatly accelerated the formation of carbonyl groups in BSA, being dependent on the concentrations of Cu(II) ions and (+)-catechin. These results suggest that Cu(II) ions could convert (+)-catechin from an antioxidant to a prooxidant in protein oxidation.
Minimal reports are available on the relationship between blood lipids such as cholesterol, triglycerides, high-density lipoprotein cholesterol (HDL-C) and low density lipoprotein cholesterol (LDL-C) and acne. Most of available literature was about the effect of drugs used in acne treatment on these parameters. In this work we determined plasma total cholesterol, triglycerides, HDL-C and LDL-C levels in 166 (83 males and 83 females) newly diagnosed untreated Jordanian acne patients and compared with 105 (52 males and 53 females) of age and sex matched healthy controls. Results indicated that acne patients, males and females, had significantly low plasma HDL-C levels (p=0.000). Plasma total cholesterol, triglycerides and LDL-C levels were shown to be within the normal range except for triglycerides and LDL-C levels in severe acne cases for both sexes, were shown to be significantly elevated compared with those in healthy controls (p=0.004 and 0.000 consequently). It has been noticed that there was a trend for plasma HDL-C of acne patients to decrease as the severity of acne condition increases. Our results indicated that acne patients have significant changes in the plasma lipids profile that should be considered in the pathogenesis as well as in the treatment of acne.
The fate of pretilachlor and esprocarb in soil with or without spherosomes was investigated for 71 days. The estimated half-lives of pretilachlor in nonsterile soil with and without spherosomes were calculated to be 35.6 and 54.3 days, respectively, while those of esprocarb in nonsterile soil with and without spherosomes were 35.5 and 44.8 days, respectively. The degradation of these pesticides appeared to be essentially due to the biological activity of the soil. Spherosomes should enhance the rate of pretilachlor and esprocarb destruction.
Rice bran was found to effectively adsorb p-dichlorobenzene. The amount of p-dichlorobenzene adsorbed was plotted against the equilibrium concentration of substsnces in solution on a logarithmic scale, and a linear relationship was obtained, indicating that the adsorption reaction was Freundlich-type. Adsorption of p-dichlorobenzene by rice bran was observed in the pH range 1-12. The removal of p-dichlorobenzene by rice bran was attributed to the uptake by intracellular particles called spherosomes.
We examined what changes occurred in the activity and content of superoxide dismutase (SOD) in the blood and the amount of 8-hydroxy-2'-deoxyguanosine (8-OHdG) in the urine as a consequence of oral administration of antioxidant health foods including plant-based SOD, vitamin C and vitamin E to seven healthy subjects every day for 15 days. Although there was a significant increase in the concentration of vitamins C and E in serum, there was a significant decrease in SOD (extracellular type) activity and Mn-SOD (mitochondrial type) content and a narrower range of variation therein. In contrast, there was a tendency toward an increase in the amount of 8-OHdG in the urine (observed in 6 of 7 subjects). We looked into the possibility that SOD activity was being inhibited by pycnogenol (water extract of the bark of the French maritime pine) as the main ingredient of the antioxidant health foods, and it became clear that SOD activity is included in pycnogenol. These results suggest that oral administration of antioxidant health foods containing SOD originating in plants has the effect of lowering the activity and content of SOD in the blood.
The traditional Japanese Kampo medicine Bushirichu-to (Bushi-ninjin-to) has been empirically used for the treatment of chronic gastroenteritis, gastric atony, and chronic constipation accompanied by a feeling of coldness in the body. One of the mechanisms of the empirical effects is assumed to be due to local changes in gut-regulated peptide levels. We studied the effect of Bushi-richu-to on calcitonin gene-related peptide (CGRP)-, substance P-, vasoactive intestinal polypeptides (VIP)-, somatostatin-, and motilin-like immunoreactive substances (IS) in plasma taken from healthy subjects. Bushi-richu-to (4.5 g) or placebo was orally administered to five healthy males. Blood samples were taken before, and at 20, 40, 60, 90, 120, 180, and 240 min after administration, extracted, and then submitted to a highly sensitive enzyme immunoassay system for CGRP-, substance P-, VIP-, somatostatin-, and motilin-IS. A single oral administration of Bushi-richu-to caused significant increases in plasma CGRP-, substance P-, VIP-, and somatostatin-IS levels compared with placebo. It is concluded that Bushi-richu-to might improve a peripheral and uncomfortable feeling of cold and gastrointestinal dysmotility via gut-regulated peptide release.
We investigated whether the efficiency of intestinal calcium (Ca) absorption was improved by concomitant ingestion of gum arabic (GA) in rats. We used the Ussing chamber method to clarify the effect of GA on upper and lower small intestinal absorption of Ca. Increased in vitro Ca permeation was observed in rats who ingested water with 7.5% GA for 10 days. These results suggested that administration of GA with Ca could increase the efficiency of oral Ca absorption.
The effect of bee pollen extract on osteoblastic MC3T3-E1 cells was investigated. The water-solubilized extracts, which were obtained from the bee pollen of Cistus ladaniferus, was purified using the membrane fractionation method with molecular weight (MW) less than 1000. Osteoblastic cells were cultured for 72 hr in a medium containing either vehicle or cistus extract of less than MW 1000 (10, 25, or 50 μg/ml of medium) in the presence of 10% fetal bovine serum (FBS). The proliferation of osteoblastic cells was significantly enhanced in the presence of cistus extract (25 or 50 μg/ml). The stimulatory effect of cistus extract on cell proliferation was also observed when the cells with subconfluency were cultured for 24 or 72 hr without FBS. Cells with subconfluency were cultured for 24 or 72 hr in a medium containing either vehicle or cistus extract without FBS. DNA content or alkaline phosphatase activity in osteoblastic cells was significantly increased after culture with cistus extract (50 μg/ml) for 24 hr. The effect of 25 μg/ml of cistus extract on these components was seen after culture for 72 hr. This study demonstrates that the cistus extract fraction of less than MW 1000 has anabolic effects in osteoblastic MC3T3-E1 cells which involve in bone formation.
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