Green tea polyphenols inhibit metalloproteinase activities in the skin,muscle, and blood of rainbow trout

We have investigated the inhibitory effects of polyphenols from natural products, such as green tea, bilberry, grape, ginkgo, and apple, on rainbow trout gelatinase activities. Gelatinases from the skin, muscle, and blood of rainbow trout contained serine proteinase, metalloproteinase, and other proteinase activities as measured by gelatin zymography. The polyphenols of green tea caused the strong inhibition of some gelatinase activities when compared with those of the other products. This inhibition was quite similar to that of metalloproteinase by ethylenediaminetetraacetic acid, suggesting that the effects of green tea polyphenols on proteinase activities are specific for metalloproteinases. The major catechins of green tea polyphenols were then separated and identified by reverse-phase chromatography to be (-)-epigallocatechin gallate (EGCG), (-)-epigallocatechin, (-)-epicatechin gallate, and (-)-epicatechin. The effects of these catechins on gelatinase activities were examined; the most potent inhibitor of metalloproteinase activities was found to be EGCG. These results have indicated that green tea polyphenols including EGCG are useful for regulating metalloproteinase activities of fish meat.     

Reverse phase liquid chromatographic determination and confirmation of aflatoxin M1 in cheese

A systematic method is proposed for determination and confirmation of aflatoxin M1 in cheese by liquid chromatography (LC). A sample of cheese is extracted with chloroform, cleaned up on 2 silica gel columns followed by a Sep-Pak C18 cartridge, and chromatographed on a 5 microns octadecyl silica column with fluorometric detection. The sample extract or standard is treated with n-hexane-trifluoroacetic acid (TFA) (4 + 1) for 30 min at 40 degrees C. Analysis by LC with TFA-treatment of the extract provides quantitative data. Multiple assays of 5 samples of Gouda cheese spiked with aflatoxin M1 at levels of 0.5, 0.1, and 0.05 ng/g showed average recoveries of 93.2, 91.6, and 92.4%, with coefficients of variation of 2.63, 3.97, and 4.52%, respectively. Assay of 5 naturally contaminated cheeses resulted in 0.051-0.448 ng/g of aflatoxin M1. Limit of quantitation is about 0.01 ng/g. The identity of aflatoxin M1 is confirmed by treating aflatoxin M1 or the M2a derivative with TFA-methanol (or ethanol) (3 + 1). The TFA-methanol reaction products of M2a could be detected quantitatively.     

Control mechanism of the circadian clock for timing of cell division in vivo

Cell division in many mammalian tissues is associated with specific times of day, but just how the circadian clock controls this timing has not been clear. Here, we show in the regenerating liver (of mice) that the circadian clock controls the expression of cell cycle-related genes that in turn modulate the expression of active Cyclin B1-Cdc2 kinase, a key regulator of mitosis. Among these genes, expression of wee1 was directly regulated by the molecular components of the circadian clockwork. In contrast, the circadian clockwork oscillated independently of the cell cycle in single cells. Thus, the intracellular circadian clockwork can control the cell-division cycle directly and unidirectionally in proliferating cells.     

Apricot carotenoids possess potent anti-amyloidogenic activity in vitro

Alzheimer's disease (AD) is the most common form of dementia and is characterized by the progressive accumulation of amyloid β protein (Aβ) in areas of the brain. There has been an increased interest in screening for food-grade anti-amyloidogenic compounds in foodstuffs. The purpose of this study was to screen and identify bioactive compounds with anti-amyloidogenicity in apricot fruits using synthetic Aβ(1-42). The anti-amyloidogenicity was investigated using thioflavin T fluorescence assay, electron microscopy, and dot blotting analysis. The carotenoid fraction from apricot showed strong inhibitory effects against oligomer and fibril formation of Aβ and fibril-destabilizing effects. Among the peaks in the HPLC chromatogram, lutein showed the strongest inhibitory effect on Aβ fibril formation. The inhibitory effect was dependent on the number and portion of hydroxyl groups on both sides of carotenoids. These findings suggest that lutein in fruits may be useful as a preventive agent for amyloid-associated diseases.     

Development of a quantification assay for the cysts of the toxic dinoflagellate <Emphasis Type="Italic">Alexandrium tamarense</Emphasis> using real-time polymerase chain reaction

ABSTRACT:   The cysts of toxic dinoflagellate Alexandrium tamarense are the seed population for the bloom responsible for paralytic shellfish poisoning (PSP). However, it is impossible to identify the Alexandrium spp. cyst on the basis of morphological features. In this study, we prepared A. tamarense cysts by sexual conjugation in laboratory conditions and developed an efficient DNA extraction method for polymerase chain reaction (PCR) assay. Using the A. tamarense cysts, we established the identification and quantification method showing the species specificity and the high sensistivity for A. tamarense cysts using real-time PCR. This assay was also able to detect and quantify the A. tamarense cysts accurately when mixed with excess cysts of A. catenella (Whedon and Kofoid) Balech prepared by conjugation experiment.     

Development of spermatogenic function in the sex maturation process in male cats

The spermatogenic function and plasma testosterone (T) level in the sex maturation process were investigated as to 180 mixed breed cats ranging from 4 months to 2 years in age to be castrated. Testis/epididymis weights reached a peak at 10 and 8 to 9 months of age, respectively. In the testis, sperm appeared at 5 months of age. At 7 months of age, sperm were observed in 96.2% of the cats. In the tail of the epididymis, sperm appeared in 46.9% of the cats at 6 months of age and in all cats at 8 or more months of age. Furthermore, the mean plasma T level rapidly increased at 8 months of age, and reached a peak (2.64 +/- 0.68 (SE) ng/ml) at 10 months of age. Three of 180 cats (1.67%) had unilateral cryptorchidism. These results suggest that the spermatogenic function in male cats becomes mature at 8 to 10 months of age.     

Quillaja saponin can modulate ovalbumin-induced IgE allergic responses through regulation of Th1/Th2 balance in a murine model

Quillaja saponin is the extract from the balk of a South American tree, and it is considered to modulate immunological responses. We hypothesized that Quillaja saponin may change allergy-associated cytokine profile and antigen-specific immune responses. The purpose of this study is to investigate whether Quillaja saponin can suppress ovalbumin (OVA)-induced IgE-mediated allergic responses through promoting a dominant Th1 immune response. The spleen cells from BALB/c mice, which were primed by OVA, were used for an in vitro challenge test. The level of total and OVA-specific IgE, IL-4, IFN-gamma, and IL-12 was determined by enzyme-linked immunosorbent assay (ELISA). BALB/c mice were orally administered with saponin for 35 days. The mice were immunized intraperitoneally with OVA on days 14 and 21. After intraperitoneal challenge with OVA on day 35, anaphylactic symptoms were monitored. Total and specific IgE and IgG, specific IgG1 and IgG2a, and histamine levels in serum were analyzed by ELISA. The increase of IL-12 and IFN-gamma levels was observed in the presence of Quillaja saponin, while the IL-4 level was decreased. Furthermore, Quillaja saponin suppressed total and OVA-specific IgE secretion in spleen cells. Balb/c mice that were orally administered Quillaja saponin exhibited lower total and OVA-specific IgE and OVA-specific IgG secretions, whereas total IgG levels remained unchanged. Suppression of OVA-specific IgG1 and an increase of OVA-IgG2a were observed in mice fed saponin. Quillaja saponin also decreased serum histamine levels and diminished anaphylactic symptoms. The present study indicates that Quillajasaponin can suppress allergen-specific IgE-mediated reactivity in a murine model of food allergy, which results from shifting from a Th2-dominated to a Th1-dominated immune response.     

Development of EST-SSR markers of Ipomoea nil

Although Japanese morning glory (Ipomoea nil (L.) Roth.) has been used intensively for genetic studies, DNA markers have not been developed in Ipomoea nil sufficient to cover all chromosomes. Therefore, we conducted microsatellite (simple sequence repeats, SSR) marker development in I. nil for future genetic studies. From 92,662 expressed sequence tag (EST) sequences, 514 unique microsatellite-containing ESTs were identified. Primer pairs were designed automatically in 326 SSRs. Of 150 SSRs examined, 75 showed polymorphisms among strains. A phenogram based on the SSR genotypes revealed the genetic relation among seven Japanese morning glories from five different regions of the world and an ivyleaf morning glory (I. hederacea Jacq.). The developed SSR markers might be applicable for genetic studies of morning glories and their relatives.