Inhibition on human liver cytochrome P450 3A4 by constituents of fennel (Foeniculum vulgare): identification and characterization of a mechanism-based inactivator

Fennel, a seed of Foeniculum vulgare, is used as a culinary spice and traditional medicine. The methanolic extract of fennel showed a characteristic of mechanism-based inactivation on erythromycin N-demethylation mediated by human liver microsomal cytochrome P450 3A4 (CYP3A4). The present study was conducted to identify the fennel constituent having the inhibition. Thirteen compounds have been isolated from a methanol extract of fennel and tested for their inhibition on CYP3A4. Among them, 5-methoxypsoralen (5-MOP) showed the strongest inhibition with an IC50 value of 18.3 microM and a mixed type of inhibition. In addition, with the preincubation time of 20 min only 5-MOP showed preincubation time dependency; the IC50 value decreased from 18.3 microM with a preincubation time of 0 min to 4.6 microM with a preincubation time of 20 min. Further investigation on 5-MOP showed the characteristics of time-dependent inhibition, requirement of NADPH, lack of protecting effect of nucleophiles, and recovery of CYP3A4 activity by the competitive inhibitor. This result suggests that the inhibitory activity of CYP3A4 by 5-MOP was a mechanism-based inactivation. The kinetic parameter for mechanism-based inactivation was characterized by a KI value of 15.0 microM and a kinact value of 0.098 min(-1).     

Supplementation of various meals to fish meal diet for chum salmon fry

In a 6-week feeding experiment, chum salmon, Oncorhynchus keta, swim-up fry, were fed fish meal diets supplemented with silkworm pupae powder (5%), dried beef liver (5%), krill meal (5%) or earthworm powder (5%) at the expense of fish meal, or substituting glucose (13%) for dextrin. Fish fed the diet with earthworm powder showed the best growth performance (675% weight gain in 6 weeks) and feed efficiency (117%). The growth rate and feed efficiency of the krill meal group were also significantly better than those of the control group (without supplement or substitution). Substitution of glucose and supplementation of silkworm pupae or beef liver failed to improve the growth rate, but significantly improved feed efficiency. Food consumption of fish fed the diet supplemented with earthworms was lowest, and none of the dietary treatments appreciably increased food intake of the fish over that of the control group. The dietary group receiving earthworm powder also showed significantly higher body fat content than the remaining dietary groups. No significant differences were noted in body protein and ash contents among all dietary treatments.     

Changes in estrogen receptor expression in the chick thymus during late embryonic development

In chickens, although estrogen receptors (ER) are reported to be associated with the immunological processes, detailed information about the differences in ER expression in the tissues related to the development of lymphocytes is not fully known, especially during the developmental stage. To learn more about this immunological relationship, we used semi‐quantitative polymerase chain reaction method to detect the ER expression levels in the thymus tissues of chicks during the developmental stage. Furthermore, ER‐expressing cells were detected by immunohistochemistry. The results of this study show that the expression level of ER increased on embryonic day 16 and decreased on day 20. Furthermore, ER expression was significantly higher in male than in female chickens at day 16. The increased expression on day 16 and decreased level on day 20 were also reproduced in the incidence of immunoreactive cells, although there was a 1‐day delay in the elevated incidence of the cells. This study revealed the changes in ER expression and the incidence of ER‐positive cells in the thymus of chickens during the developmental stage.     

Comprehensive Analysis of Plant Gene Expression in Soybean Root Nodules at Different Growth Stages

Gene expression profiles in early (12 d after sowing, DAS), mature (35 DAS) and senescent (77 DAS) soybean ( Glycine max (L.) Merr. cv. Enrei) root nodules were analyzed using Lotus japonicus cDNA macroarray. The cDNA macroarray membranes contained 18,144 independent cDNAs of L. japonicus isolated from various organs. The membranes were hybridized to ³³P-labeled single-strand cDNAs transcribed from mRNA extracted from the soybean root nodules at three different growth stages. The results showed that over 75% of the cDNA clones gave a relative signal intensity (RSI) lower than 300, irrespective of the soybean growth stage. cDNA with an RSI value over 5,000 accounted for 2.1, 1.2 and 0.5% of the total cDNA clones at 12, 35 and 77 DAS, respectively. The highest RSI value was detected in a clone hybridized to putative pectinesterase (MWM097c10_r) at 12 DAS, while the RSI value of lycopene β-cyclase (MWL025d06_r) was the highest at 35 and 77 DAS. At 77 DAS, the percentage of transferase gene cDNA clones accounted for 20.5% of the total cDNA clones showing an RSI value over 500, a value clearly lower than that at 12 and 35 DAS. On the other hand, the percentage of hydrolase gene cDNA clones accounted for 45.2% at 77 DAS, a value around 1.5 times higher than that at 12 and 35 DAS. The chronological RSI profiles of individual cDNA clones were classified into nine patterns, and the profiles of the cDNA clones belonging to 15 homologous gene groups encoding enzymes of nitrogen and carbon metabolism, nodule specific proteins, enzymes of RNA and protein hydrolysis, and synthesis of jasmonic acid precursor were compared within and between groups. As a result, significant variations both within- and between-groups were revealed.     

Identification of food-derived collagen peptides in human blood after oral ingestion of gelatin hydrolysates

In the present study, we identified several food-derived collagen peptides in human blood after oral ingestion of some gelatin hydrolysates. Healthy human volunteers ingested the gelatin hydrolysates (9.4-23 g) from porcine skin, chicken feet, and cartilage after 12 h of fasting. Negligible amounts of the peptide form of hydroxyproline (Hyp) were observed in human blood before the ingestion. After the oral ingestion, the peptide form of Hyp significantly increased and reached a maximum level (20-60 nmol/mL of plasma) after 1-2 h and then decreased to half of the maximum level at 4 h after the ingestion. Major constituents of food-derived collagen peptides in human serum and plasma were identified as Pro-Hyp. In addition, small but significant amounts of Ala-Hyp, Ala-Hyp-Gly, Pro-Hyp-Gly, Leu-Hyp, Ile-Hyp, and Phe-Hyp were contained.