Role of metal proteinases in the fruit-body formation ofHypsizygus marmoreus

We investigated the possible role of metal proteinase on the fruit-body formation ofHypsizygus marmoreus. The addition of a specific metal proteinase inhibitor, phosphoramidon, to the culture medium (10g/ml) completely inhibited fruit-body formation. Metal proteinase activity in both the medium and the mycelia of this fungus increased markedly during vegetative mycelial growth, and activity was maximal 25 days after inoculation. When phosphoramidon was added to the culture medium during vegetative mycelial growth, the metal proteinase activity in the mycelium decreased to 56% of the control (without inhibitor) level. Isoelectric focusing analysis showed that two kinds of metal proteinases with a pl of 7.7 and 8.4, respectively, were obtained from 29-day-old mycelia. Uptake of phosphoramidon into the mycelia was confirmed as the result of inhibition of thermolysin activity by the mycelial extracts. The degree of inhibitor uptake into mycelia was about 2.0% and was independent of the initial concentration of the inhibitor administered. The addition of peptone and amino acids to medium treated with phosphoramidon resulted in fruit-body dry weight yields that were about 50% that of the control.     

Antioxidative and antimutagenic activities of 4-vinyl-2,6-dimethoxyphenol (canolol) isolated from canola oil

A potent antioxidative compound in crude canola oil, canolol, was recently identified, and reported herein are studies of its scavenging capacity against the endogenous mutagen peroxynitrite (ONOO(-)). ONOO(-) is generated by the reaction between superoxide anion radical and nitric oxide, both of which are produced by inflammatory leukocytes. Among various antioxidative substances of natural or synthetic origin, canolol was one of the most potent antimutagenic compounds when Salmonella typhimurium TA102 was used in the modified Ames test. Its potency was higher than that of flavonoids (e.g., rutin) and alpha-tocopherol and was equivalent to that of ebselen. Canolol suppressed ONOO(-)-induced bactericidal action. It also reduced intracellular oxidative stress and apoptosis in human cancer SW480 cells when used at a concentration below 20 microM under H(2)O(2)-induced oxidative stress. In addition, canolol suppressed plasmid DNA (pUC19) strand breakage induced by ONOO(-), as revealed by agarose gel electrophoresis.     

Inhibitory actions of several natural products on proliferation of rat vascular smooth muscle cells induced by Hsp60 from Chlamydia pneumoniae J138

Atherosclerosis is a vascular disorder involving inflammation, a narrowed vascular lumen in the entire tunica intima, and reduced elasticity of the arterial wall. It has been found that Hsp60 from Chlamydia pneumoniae, an obligate bacterial pathogen associated with atheroma lesions, mimics human Hsp60, thereby causing attacks by immune cells on stressed endothelial cells expressing endogenous Hsp60 on their surface. Furthermore, Hsp60 from C. pneumoniae has been shown to promote the growth of vascular smooth muscle cells (VSMCs). To explore probes that can be used for studying signal transduction elicited by the chlamydial Hsp60, we have tested several natural products for their inhibitory actions on the Hsp60-induced proliferation of rat arterial smooth muscle cells. Sesamol, vanillyl alcohol, and trans-ferulic acid exhibited moderate inhibitory actions on the Hsp60-induced cell proliferation; zerumbone, humulene, and caryophylene effectively inhibited it at low concentrations with IC(50) values of 529, 122, and 110 nM, respectively. The results indicated that the 11-membered alicyclic ring is favorable for interactions with receptors involved in the Hsp60-induced VSMC proliferation.     

Effect of pH Adjustment on Preservation of Kitchen Waste Used for Producing Lactic Acid

Interest in the production of lactic acid (LA) is increasing inrelation to its applications in the synthesis of biodegradablepolymer materials. Before kitchen waste is used as substrates inLA fermentation, it must be stored for some days in the wastecollection system. This study examined the effects of pHadjustment including initial pH before the storage in terms of thekind and the amount of added acid on preservation of the waste. The optical purity of LA generated during the storage was alsoanalyzed. It was found that antibacterial activity andsuppression ability to D-lactic acid (D-LA) were stronger in theorder: acetic acid > L-lactic acid (L-LA) > hydrochloric acid ≈ nitric acid. The optimum condition of waste storage was found to be adjusting initial pH to 3.4 by adding acetic acid. It can be expected that preservation of kitchen waste can be realized by inhibiting the growth of putrefactive bacteria, and higher optical purity of L-LA was obtained by adding weak acid to the waste.     

Physical properties of shark gelatin compared with pig gelatin

Physical properties of shark gelatin were examined during gel formation and postgelation in comparison with pig gelatin. Samples with various concentrations and pH values were evaluated by breaking strength, dynamic viscoelasticity, and dynamic light scattering. Sol-gel and gel-sol transition temperatures for shark gelatin were remarkably lower than those for pig gelatin. Shark gelatin gel shows a narrower pH range to form a stable gel compared with pig gelatin. Melting enthalpy of shark gelatin gel was greater than that of pig gelatin gel, and G' of shark gelatin gel changed more extensively with rising temperature in comparison with pig gelatin gel. It is concluded that shark gelatin has different characteristics from pig gelatin not only for gel characteristics but also for the solution property.     

Ultrastructure of the oocytes of the Japanese eel Anguilla japonica during artificially induced sexual maturation

ABSTRACT: Morphological changes in the oocytes of the Japanese eel, Anguilla japonica , induced to undergo ovarian development by repeated injections of salmon pituitary homogenate, were examined using electron microscopy. Oil droplets were closely associated with organelles, especially mitochondria, and increased in number as oocyte growth proceeded. They fused at the migratory nucleus stage. During vitellogenesis, two types of cortical alveoli were distinguished, one having filamentous contents, the other having latticated contents. As oocytes reached maturity, the structure of the cortical alveoli was exclusively filamentous. Yolk globules were homogeneous and highly electrondense, but electrondensity decreased during hydration. The structure of the zona radiata of previtellogenic oocytes consisted of two layers, and an additional reticular network structure was formed on the inside of the zona radiata during the vitellogenic stage. The zona radiata lost the reticular network structure and assumed a layered structure of uniform electrondensity at the migratory nucleus stage. These structural changes during oocyte development were mostly comparable to those in other teleosts. Results of the present study should assist in developing improved methods for full control of artificial maturation in the Japanese eel.     

Expression patterns of the slit subfamily mRNA in canine malignant mammary tumors

Slit, a secreted protein, functions as a chemorepellent factor in axon guidance and neuronal migration and as an inhibitor in leukocyte chemotaxis. In humans, slit2 protein attracts endothelial cells and promotes tube formation in the tumor angiogenic mechanism. In this study, we cloned a part of the canine slit subfamily and examined the expression of slit subfamily mRNAs in 3 normal canine mammary glands and 11 mammary tumor samples by RT-PCR. The cloned part of the slit gene sequences showed high similarity to those of the human, mouse, and rat. The mRNAs were expressed at low levels in the normal mammary gland. The expression levels of slit1 mRNA were low in both the normal and tumor tissues. In contrast, the expression of slit2 mRNA increased in most of the malignant mammary tumors, and an increase in slit3 mRNA expression was observed in 2 of the malignant mixed tumors. These results suggest that the expression of slit2 plays an important role in tumor angiogenesis in canine mammary gland tumors and that slit2 can be a putative marker for malignancy diagnosis of these tumors.     

Evaluation by weight change rate of dairy herd condition

The purpose of this study was to clarify whether weight change rate (WCR) is useful in evaluating herd health. Five herds (A, B, C, D, and E) were examined monthly for approximately 1 year to measure both WCR and BCS. WCR was calculated using the following equation: WCR (%) = (body weight of current month-body weight of the preceding month)/body weight of current month x 100. In addition, blood profiles were performed in each herd before and after calving. The incidence of periparturient diseases was higher in Herds A, B, and C compared with Herds D and E. Reproductive performance in Herds A and B was rather poor compared with Herds C, D, and E. There were significantly lower WCRs in Herd A compared with Herd C during the period of day -30 to day 30 after calving. According to the blood profiles, high levels of non-essential fatty acids (NEFA) were observed during the period of day -30 to -1 and high beta-hidroxybutyricacid (BHB) and low levels of glucose were observed at day 30 after calving in Herd A. A marked poor condition in Herd A during the peripaturient period was clearly expressed by low WCR in this experiment. WCR can express body condition of herds and is suggested to be a useful parameter in the determination of herd performance in field experiments.