Detection of antifungal activity in Anemarrhena asphodeloides by sensitive BCT method and isolation of its active compound

Antifungal activity was detected from Anemarrhena asphodeloides by the Bio-Cell Tracer (BCT) method. An active fraction was separated by silica gel column chromatography and reverse-phase HPLC. The molecular weight was determined by GC-MS, and the molecular structure was analyzed by IR, (1)H NMR, and (13)C NMR. The isolated compound was found to be identical to nyasol, (Z)-1, 3-bis(4-hydroxyphenyl)-1,4-pentadiene, which formerly appeared in the literature without any remark on the antifungal activity. This compound showed antimicrobial activity against 38 strains of fungi and five strains of bacteria. The minimum inhibitory concentration (MIC) ranged from 12.5 to 200 microg mL(-)(1), except for two strains based on the broth dilution method.     

Effects of cooking on concentrations of polychlorinated dibenzo-p-dioxins and related compounds in fish and meat

We investigated the cooking-induced changes in concentrations of polychlorinated dibenzo-p-dioxins (PCDDs), polychlorinated dibenzofurans (PCDFs), and dioxin-like polychlorinated biphenyls (PCBs) (dioxins) using mackerel and beef. The concentrations of dioxins (29 congeners) were determined by isomer specific analyses and were compared between uncooked and cooked samples. The cooking procedures examined in this study included grilling as a fillet, boiling as a fillet, and boiling as tsumire (small, hand-rolled balls) for mackerel and boiling as a slice, broiling as a slice, and broiling as a hamburger for beef. Three trials were carried out for each cooking method. Generally, concentrations of dioxins were reduced in every cooking trial. When nondetected congener concentrations were assumed to be half the limit of detection for mackerel, the maximum percentage reductions of total concentrations given as 2,3,7,8-tetraCDD equivalents (TEQ) were 31% in grilling as a slice, 14% in boiling as a slice, and 21% in boiling as tsumire under the conditions of this study. In contrast, for beef, the reductions were 42% in boiling as a slice, 42% in broiling as a slice, and 44% in broiling as a hamburger. These results suggest that ordinary cooking processes with heating undoubtedly reduce the dioxin content in animal products, and the reductions estimated should be considered when dioxin intake is evaluated using contamination data for individual food items.     

Flavonol Caffeoylglycosides as alpha-Glucosidase Inhibitors from Spiraea cantoniensis Flower

In the screening experiments for rat intestinal alpha-glucosidase inhibitors in 218 plants cultivated in the Japanese temperate region, potent maltase-inhibiting activity was found in the extract of flowers of Spiraea cantoniensis. The enzyme assay guided fractionation of the extract led to the isolation of three flavonol caffeoylglycosides, quercetin 3- O-(6- O-caffeoyl)-beta-galactoside ( 1), kaempferol 3- O-(6- O-caffeoyl)-beta-galactoside ( 2), and kaempferol 3- O-(6- O-caffeoyl)-beta-glucoside ( 3), as rat intestinal maltase inhibitors. This is the first report on the alpha-glucosidase-inhibitory activity of those flavonol caffeoylglycosides. Comparison in the activity of the isolates indicated the importance of caffeoyl substructures in the molecule for the alpha-glucosidase-inhibiting activity. The relatively high contents of the active isolates in the plant suggest that S. cantoniensis could be physiologically useful for treatment of diabetes.     

The first occurrence of leaf mold of tomato caused by races 4.9 and 4.9.11 of <Emphasis Type="Italic">Passalora fulva</Emphasis> (syn. <Emphasis Type="Italic">Fulvia fulva</Emphasis>) in Japan

Tomato leaf mold caused by Passalora fulva was found on two tomato varieties carrying the Cf-9 gene in Japan, in 2007. The isolates obtained from Chiba and Fukushima were identified as race 4.9.11, and those from Gunma were races 4.9 or 4.9.11. This is the first report in Japan of tomato leaf mold caused by P. fulva strains that can overcome the Cf-9 gene.     

Effect of the span length of Granier-type thermal dissipation probes on sap flux density measurements

? Granier-type thermal dissipation sensors measure sap flux density (u) by using the temperature difference between the heater and the reference probe. To detect u correctly, heat must not be transferred to the reference probe by thermal conduction. The distance across which heat can be transferred by conduction is important for the span length of a sensor and spacing of a number of sensors.

? To validate span lengths and spacing of sensors, we used numerical simulations to calculate the potential distance across which heat can be transferred by conduction. We compared measurements with an original and a modified sensor for a Japanese red pine (Pinus densiflora) from December 2004 to May 2005. The span length of the original and the modified sensor is 15 and 4 cm, respectively.

? Numerical simulations showed that span length and spacing of Granier sensors should be more than 10 cm for trees in which u ceases for a few hours before the predawn period. The modified sensor underestimated u by 18–46% in winter (December–March) because its reference temperature was increased by heat transferred by conduction. The modified sensor measured u correctly in warm seasons, and only underestimated the annual amount of transpiration by 6%.

     

Distribution of the introduced cyprinid herpesvirus 3 in a wild population of common carp, Cyprinus carpio L.

Cyprinid herpesvirus 3 (CyHV-3), which causes a lethal disease in common carp, Cyprinus carpio L., and koi, C. carpio koi , first occurred in Lake Biwa, Japan in 2004. To elucidate distribution of CyHV-3 in a wild common carp population, we conducted a PCR survey of CyHV-3 among such fish in Lake Biwa in 2006. Only 6% (1/18) of the common carp smaller than 300 mm were positive with PCR, whereas 31% (18/58) of fish larger than 300 mm were positive. To evaluate their past exposure to CyHV-3 infection based on the presence of antibodies, we also measured the levels of serum anti-CyHV-3 antibodies in the carp, using an enzyme-linked immunosorbent assay. None (0/26) of the fish smaller than 300 mm was positive for the antibodies, whereas 54% (33/61) of fish larger than 300 mm were positive. Of the antibody-positive individuals, 44% (14/32) were also positive by PCR strongly suggesting that wild common carp that survived infection become CyHV-3 carriers. Five individuals were positive by PCR but negative for antibodies indicating that their infection with CyHV-3 had occurred recently. These results suggest that transmission of CyHV-3 from carriers to naïve common carp is still occurring in Lake Biwa.     

Effect of exposure to low salinity water on plasma ion regulation and survival rates in artificially wounded devil stinger <Emphasis Type="Italic">Inimicus japonicus</Emphasis>

We investigated the effect of exposure to low salinity water on plasma ion regulation and survival rates in artificially wounded devil stinger Inimicus japonicus. All fishes survived in 33% seawater (SW), while survival rate in 100% SW was 5.1% at 24 h. In 100% SW, plasma Na⁺, K⁺, Mg²⁺, and Ca²⁺ concentrations significantly increased to 238?±?49.9, 9.6?±?2.4, 15.1?±?3.5 and 5.0?±?0.7 mmol/l at 6 h, respectively; the gill Na⁺/K⁺–ATPase (NKA) activity was almost stable, although only one fish survived to 24 h. In 33% SW, plasma Na⁺ and K⁺ concentrations remained at the same level, and plasma Mg²⁺ and Ca²⁺ concentrations gradually increased to 16.2?±?0.7 and 4.5?±?0.2 mmol/l until 24 h, respectively. The NKA activity significantly increased to 5.1?±?1.1 µmol ADP/mg protein per h at 6 h. A positive correlation was observed between the wound surface area against body weight and the plasma ion concentrations, although no difference was observed in the restoration rate of the wounded area between 100 and 33% SW. These results indicate that exposure of wounded fish to low salinity water improves survivability by favoring plasma ion regulation without influencing the restoration rate.     

Plasticity of animal genome architecture unmasked by rapid evolution of a pelagic tunicate

Genomes of animals as different as sponges and humans show conservation of global architecture. Here we show that multiple genomic features including transposon diversity, developmental gene repertoire, physical gene order, and intron-exon organization are shattered in the tunicate Oikopleura, belonging to the sister group of vertebrates and retaining chordate morphology. Ancestral architecture of animal genomes can be deeply modified and may therefore be largely nonadaptive. This rapidly evolving animal lineage thus offers unique perspectives on the level of genome plasticity. It also illuminates issues as fundamental as the mechanisms of intron gain.     

Optimization of a protocol for cryopreservation of mouse spermatozoa using cryotubes

The rapid increase in the number of genetically modified mouse strains has produced a high demand for their frozen spermatozoa from laboratories and mouse banking facilities. Historically, plastic straws have been used preferentially as containers for frozen mammalian spermatozoa because spermatozoa frozen in plastic straws have a high survival rate after thawing. However, plastic straws are more fragile and are used less often than the cryotubes used for conventional cell freezing. In this study, we sought to develop a new protocol for sperm freezing using cryotubes as the container to increase the accessibility of mouse sperm cryopreservation. Epididymal spermatozoa were collected from mature ICR or C57BL/6J (B6) males and were suspended in 18% raffinose and 3% skim milk solution. We then optimized the following conditions using the sperm survival rate as an index: 1) distance of cryotubes from the surface of the liquid nitrogen at freezing, 2) volume of the sperm suspension in the cryotube and 3) temperature of warming sperm during thawing. The best result was obtained when cryotubes containing 10 μl of sperm suspension were immersed 1 cm below the surface of the liquid nitrogen and then thawed at 50 C. The fertilization rates using spermatozoa frozen and thawed using this method were 63.1% in ICR mice and 28.2% in B6 mice. The latter rate was increased to 62.3% by adding reduced glutathione to the fertilization medium. After embryo transfer, 68% and 62% of the fertilized oocytes developed into normal offspring in the ICR and B6 strains, respectively. These results show that cryotubes can be used for cryopreservation of mouse spermatozoa under optimized conditions. This protocol is easy and reproducible, and it may be used in laboratories that do not specialize in sperm cryopreservation.