Kale is a cruciferous vegetable (Brassicaceae) that contains a large amount of health-promoting phytochemicals. The chronic ingestion of cabbage of the same family is known to accelerate conjugating acetaminophen (AA) and decrease the plasma AA level. Therefore, we examined to clarify the effects of kale on the pharmacokinetics of AA, its glucuronide (AA-G) and sulfate (AA-S). AA was orally administered to rats pre-treated with kale or cabbage (2000 mg/kg/day) for one week. Blood samples were collected from the jugular vein, and the concentrations of AA, AA-G and AA-S were determined. In results, kale ingestion induced an increase in the area under the concentration-time curve (AUC) and a decrease in the clearance of AA, whereas cabbage had almost no influence. In addition, there were significant differences in the AUC of AA-G between the control and kale groups. mRNA expression levels of UDP-glucuronosyltransferases, the enzymes involved in glucuronidation, in the kale group were significantly higher than those in the control group. In conclusion, kale ingestion increased the plasma concentrations of both AA and AA-G. The results suggest that kale ingestion accelerates the glucuronidation of AA, but an increase of plasma AA levels has a different cause than the cause of glucuronidation.
This study investigated whether the suppression of hypoxia-inducible factor (HIF)-1α up-regulation prevents high-dose and long-term UVB-induced wrinkle formation and angiogenesis associated with increased matrix metalloproteinase (MMP)-2 and MMP-9 activities. Twenty-four hairless mice were assigned to three groups: 1) control, 2) UVB-irradiation (UVB), and 3) UVB-irradiation followed by hyperoxia (UVB+HO). The backs of the mice were exposed to UVB irradiation three times a week for 10 weeks. To suppress UVB-induced cutaneous HIF-1α up-regulation, the mice were exposed to hyperoxia (90% oxygen) for 2 h immediately after each UVB irradiation. The UVB and UVB+HO groups had significantly increased degrees of wrinkle formation, dermal blood vessel density, and MMP-2 and MMP-9 activities compared with the control group. HIF-1α expression levels were significantly higher in the UVB group than in the control group whereas these levels remained unchanged in the UVB+HO groups. The activity of type 1 collagenase which digests collagen type 1 (a main component of the dermis), was similar in all groups; furthermore, the dermal soluble collagen content was similar in all groups. These results suggest that the suppression of increases in HIF-1α levels alone is insufficient to restrain wrinkle formation caused by higher doses and longer periods of the UVB irradiation that led to the up-regulation of MMP-2 and MMP-9 activities.
Testican, a chondroitin/heparan sulfate proteoglycan, is primarily expressed in neurons of the adult and embryonic mouse brain, suggesting its role in normal and/or proliferation and differentiation processes of neurons. However, the role of testican in injured brain remains unclear. In the present study we investigated testican-1 mRNA expression pattern after cryo-injury of the brain. In situ hybridization histochemistry revealed that testican-1 mRNA is induced in the region surrounding the necrotic tissue. Time course study of testican-1 mRNA showed the highest level of signal intensity at 7 days after the injury. To determine which cell types express testican-1 mRNA, we performed in situ hybridization histochemistry combined with immunohistochemistry of several cell markers. Testican-1 mRNA signals were detected in the proximal reactive astrocytes, whereas the distribution pattern of testican-1 mRNA positive cells was different from those of mature oligodendrocytes and activated microglia. In addition, signals for testican-1 mRNA overlapped with those of FGF-2 mRNA, showing that these molecules are coexpressed in reactive astrocytes. These results suggest a possibility that testican-1 plays a permissive role for regenerating axons in reactive astrocytes after injury.
Caspase-8 and caspase-9 play crucial roles in the extrinsic and intrinsic apoptotic pathways, respectively. The nuclear translocation of apoptosis-inducing factor (AIF) is involved in caspase-independent apoptosis. Microtubule-associated protein 1 light chain 3 (LC3) plays a pivotal role in autophagy. In the present study, we analyzed the expression of cleaved caspase-8 (CC8), cleaved caspase-9 (CC9), AIF, and LC3 in 160 gastrointestinal adenocarcinomas. The nuclear expression of AIF was rare. The expression of CC8 in gastric and colorectal adenocarcinomas did not differ, whereas the percentage of CC9-positive tumors in gastric adenocarcinomas was significantly higher than in colorectal adenocarcinomas. In contrast, the percentage of LC3-positive tumors in gastric adenocarcinomas was significantly lower than in colorectal adenocarcinomas. CC8 and CC9 occasionally co-existed in the same tumor cells in gastric adenocarcinoma. However, LC3-positive tumor cells in colorectal adenocarcinomas were constantly negative for CC8. No correlation was identified between the expression of any markers and clinicopathological parameters. These results suggest that different cell death pathways are activated in a manner that depends upon the primary site and cell type. The extrinsic and intrinsic apoptotic pathways may be mutually regulated in gastric adenocarcinomas. Also, autophagy may function as a cellular guardian to avoid apoptosis in colorectal adenocarcinomas.
The aim of this study was to determine the effects of dietary supplementation with a combination of fish oil, bilberry extract, and lutein on subjective symptoms of asthenopia in humans by a double- blind, randomized, parallel-group, and placebo-controlled trial. In the Active group, eleven subjects ingested a supplement containing omega-3 fatty acid-rich fish oil (docosahexaenoic acid 783 mg/day, eicosapentaenoic acid 162 mg/day), bilberry extract (anthocyanidin 59 mg/day), and lutein (17.5 mg/day) in soft gel capsule form, every day for 4 weeks. In the Placebo group, nine subjects ingested placebo capsules. Before and after supplementation, subjects completed a questionnaire to determine their asthenopia symptoms and were also assessed for mental fatigue symptom by the visual analog scale (VAS) test. Asthenopia symptoms such as “stiff shoulder, low back pain”, “frustration”, “dry-eye”, and “stuffy head” were improved in the Active group. Furthermore, a score of mental fatigue was improved after 4 weeks of supplementation, and no side effects were observed after the 4-week supplementation and a 2-week washout period in the Active group. These results suggest that dietary supplementation with the combination of omega-3 fatty acid-rich fish oil, bilberry extract, and lutein may safely improve subjective symptoms of asthenopia and mental fatigue in humans.
The objective of this study was to examine the association between sleep bruxism and psychological stress. The subjects consisted of 76 volunteers, who were divided into those with and without bruxism according to the diagnostic criteria for sleep bruxism outlined by the American Academy of Sleep Medicine (AASM). Stress sensitivity was evaluated before and after an experimental stress task, which involved simple mathematical calculations. It was assessed objectively by measuring the subjects' salivary chromogranin A (CgA) levels and subjectively using a ten-division visual analog scale (VAS). Compared with those observed before the stress task, the mean salivary CgA levels of the non-bruxism group (n = 54) were not significantly increased after the stress task. Conversely, the mean salivary CgA levels of the bruxism group (n = 22) were significantly increased after the stress task (P < 0.01). The mean VAS scores of the groups without (n = 54) and with (n = 22) bruxism were significantly (P < 0.01) increased after the stress task compared with those observed before the stress task, but no differences were detected between the two groups in the stress task. These findings suggest that there is an association between sleep bruxism and psychological stress sensitivity.
Gallstone disease is one of the most prevalent digestive diseases. The frequency of gallstone disease is about 10% in middle-age persons and 20% in aged persons. Gallbladder dysmotility and stasis of bile flow promote sludge and/or gallstone formation. Gallbladder contraction depends on cholecystokinin (CCK) via CCK-1 receptors (R)s. Previously, we raised CCK-1R deficient (minus;/minus;) mice and observed sludge and/or gallstone formation in more than 30% at 12-24 months of age. As ursodeoxycholate (UDCA) is commonly used for patients with gallstone disease, we expected that continuous administration of UDCA could prevent sludge and/or gallstone formation in CCK- 1R(minus;/minus;) mice. A diet containing 0.1% UDCA was administered in age-matched CCK-1R(minus;/minus;) and wild-type male and female mice for 8 months. Administration of UDCA decreased the frequency of sludge and/or gallstone formation compared with the control (CRF-1) diet (39%→26% in male, 35%→25% in female mice); however, these effects did not attain a level of statistical significance. Although the body weight was significantly higher in UDCA-fed than CRF-1-fed male mice regardless of genotypes, the plasma lipid concentrations did not differ between the two diets. In conclusion, administration of UDCA was less effective than expected at preventing sludge and/or gallstone formation in CCK-1R(minus;/minus;) mice.
Angiotensin I-converting enzyme (ACE, EC 18.104.22.168) is one of the most important enzymes in the renin-angiotensin system, a major blood pressure control system in mammals. We synthesized novel internally quenched fluorogenic (IQF) substrates for ACE based on the cleavage site of an angiotensin I, introducing N-methyl anthranic acid (Nma) and Nε-2,4-dinitrophenyl-lysine (Lys(Dnp))at the N- and C-terminal regions. Kinetic parameters of the synthesized IQF substrates Nma-Phe-His-Lys(Dnp) and Nma-His-Pro-Phe-Lys(Dnp)-Pro were compared with those of a common peptide substrate for ACE, hippuryl (Hip)-His-Leu. The kcat/Km values of Nma-Phe-His-Lys(Dnp), Nma-His-Pro-Phe-Lys(Dnp)-Pro, and Hip-His-Leu were 5.12, 1.90, and 0.80 μM-1 s-1 for rabbit lung ACE, and 16.0, 7.36, and 0.30 μM-1 s-1 for recombinant human (rh)-ACE, respectively. These results indicate that Nma-Phe-His-Lys(Dnp) is an excellent substrate for rh-ACE. Carboxypeptidase Y also hydrolyzed Nma-Phe-His-Lys(Dnp) efficiently with Km, kcat, and kcat/Km values of 60.2 μM, 105 s-1, and 1.74 μM-1 s-1, respectively. On the other hand, carboxypeptidase B did not hydrolyze IQF substrates. The newly developed IQF substrate, Nma-Phe-His-Lys(Dnp), is a valuable tool for ACE and carboxypeptidase studies.