Phosphorylation of inositol 1,4,5-triphosphate receptor 1 during in vitro maturation of porcine oocytes

During fertilization in mammalian species, a sperm-induced intracellular Ca²⁺ signal ([Ca²⁺]_i) mediates both exit of meiosis and oocyte activation. Recently, we demonstrated in mouse oocytes that the phosphorylation levels of inositol 1,4,5 trisphosphate receptor type1 (IP₃R1), the channel responsible for Ca²⁺ release and oscillations during fertilization, changed during maturation and fertilization. Therefore, we examined the expression and phosphorylation of IP₃R1 during in vitro maturation of pig oocytes. Here, our present study shows that expression of IP₃R1 protein did not change during maturation, although the phosphorylation status of the receptor, specifically at an MPM-2 epitope, did. We found that while at the beginning of maturation IP₃R1 lacked MPM-2 immunoreactivity, it became MPM-2 reactive by 24 h and reached maximal reactivity by 36 h. Interestingly, the acquisition of MPM-2 reactivity coincided with the activation of p34^cdc2 kinase and mitogen-activated protein kinase (MAPK), which are involved in meiotic progression. Following completion of maturation, inactivation of MAPK by U0126 did not affect IP₃R1 phosphorylation, although inactivation of p34^cdc2 kinase by roscovitine dramatically reduced IP₃R1 phosphorylation. Neither inhibitor affected total expression of IP₃R1. Altogether, our results show that IP₃R1 undergoes dynamic phosphorylation during maturation and this might underlie the generation of oscillations at fertilization.     

Simultaneous detection by multiplex PCR of the high-temperature-growing Pythium species: P. aphanidermatum, P. helicoides and P. myriotylum

The objective of this study was to develop a multiplex PCR detection method for the high-temperature-growing pathogens Pythium aphanidermatum, P. helicoides and P. myriotylum. Species-specific primer pairs were designed that targeted the rDNA ITS regions. The multiplex PCR was constructed with a universal primer pair for eukaryotes directed at the 18S rDNA as a positive control, in addition to the three species-specific primer pairs. When the multiplex PCR was applied to naturally infested soils, the expected species were reliably identified, suggesting that the method is suitable for the detection of the three Pythium pathogens in environmental samples.     

Evaluation of bone toxicity in various bones of aged rats

The aim of the present study was to provide a method for evaluating bone toxicity induced by drugs in various bones in aged rats. Male Crl:CD (SD) rats at 46 weeks of age were administered 15 mg/m(2) body surface area of doxorubicin, which effects the growth plate in weanling rats, weekly for 9 weeks by intravenous injection, and the femur, sternum, humerus and tibia were examined histopathologically. In the doxorubicin-treated group, thinning of the growth plate was remarkably observed in the proximal tibia and humerus; however, these changes were not observed in other regions. In addition, the osteoclast number per bone perimeter in the proximal tibia was significantly higher than others in control aged rat. Thus, recognizing the various histological reactions related to the time of epiphyseal closure is important for evaluating bone toxicity in aged rats.     

Inhibition of capsular protein synthesis of Pasteurella multocida strain P-1059

A mutant strain, PBA322, was constructed by electroporation of a phagemid containing the coding region of antisense RNA of the ompH gene, encoding 39 kDa capsular protein or OmpH, into the parental strain P-1059 (serovar A:3) of Pasteurella multocida, and the pathogenicity was determined in mice and chickens. Grayish colonies of the mutant, indicating loss of capsule synthesis, were observed under a stereomicroscope using obliquely transmitted light, while iridescent colonies were observed for the parental strain. Moreover, strain PBA322 showed a low amount of OmpH compared with the parental strain on SDS-PAGE. Additionally, the capsule of strain PBA322 was thinner than that of the parental strain according to electron microscopy, correlating to the attenuation against chickens. In conclusion, strain PBA322, the mutant of P. multocida strain P-1059, was completely attenuated for chickens.     

Herbivore-induced volatiles from tea (Camellia sinensis) plants and their involvement in intraplant communication and changes in endogenous nonvolatile metabolites

As a defense response to attacks by herbivores such as the smaller tea tortrix ( Adoxophyes honmai Yasuda), tea ( Camellia sinensis ) leaves emit numerous volatiles such as (Z)-3-hexen-1-ol, linalool, α-farnesene, benzyl nitrile, indole, nerolidol, and ocimenes in higher concentration. Attack of Kanzawa spider mites ( Tetranychus kanzawai Kishida), another major pest insect of tea crops, induced the emission of α-farnesene and ocimenes from tea leaves. The exogenous application of jasmonic acid to tea leaves induced a volatile blend that was similar, although not identical, to that induced by the smaller tea tortrix. Most of these herbivore-induced plant volatiles (HIPV) were not stored in the tea leaves but emitted after the herbivore attack. Both the adaxial and abaxial epidermal layers of tea leaves emitted blends of similar composition. Furthermore, HIPV such as α-farnesene were emitted mostly from damaged but not from undamaged leaf regions. A principal component analysis of metabolites (m/z 70-1000) in undamaged tea leaves exposed or not to HIPV suggests that external signaling via HIPV may lead to more drastic changes in the metabolite spectrum of tea leaves than internal signaling via vascular connections, although total catechin contents were slightly but not significantly increased in the external signaling via HIPV.     

Effects of Zeolite on Soil Nutrients and Growth of Barley Following Irrigation with Saline Water

ABSTRACT

Soil salinity is a major abiotic factor limiting crop production but an amendment with synthetic zeolite may mitigate effects of salinity stress on plants. The objective of the study was to determine the effects of zeolite on soil properties and growth of barley irrigated with diluted seawater. Barley was raised on a sand dune soil treated with calcium type zeolite at the rate of 1 and 5% and irrigated every alternate day with seawater diluted to electrical conductivity (EC) levels of 3 and 16 dS m^?1. Irrigation with 16 dS m^?1 saline water significantly suppressed plant height by 25%, leaf area by 44% and dry weight by 60%. However, a substantial increase in plant biomass of salt stressed barley was observed in zeolite-amended treatments. The application of zeolite also enhanced water and salt holding capacity of soil. Post-harvest soil analysis showed high concentrations of calcium (Ca^{2 +}), magnesium (Mg^{2 +}), sodium (Na⁺), and potassium (K⁺) due to saline water especially in the upper soil layer but concentrations were lower in soils treated with zeolite. Zeolite application at 5% increased Ca^{2 +} concentration in salt stressed plants; concentrations of trace elements were also increased by 19% for iron (Fe^{2 +}) and 10% for manganese (Mn^{2 +}). The overall results indicated that soil amendment with zeolite could effectively ameliorate salinity stress and improve nutrient balance in a sandy soil.     

Field trap test for bioassay of synthetic (1<Emphasis Type="Italic">S</Emphasis>,4<Emphasis Type="Italic">R</Emphasis>)-4-isopropyl-1-methyl-2-cyclohexen-1-ol as an aggregation pheromone of <Emphasis Type="Italic">Platypus quercivorus</Emphasis> (Coleoptera: Platipodidae)

An ambrosia beetle, Platypus quercivorus, is a vector of Japanese oak wilt, which causes massive mortality of oak trees in Japan. Previous studies have identified (1S,4R)-4-isopropyl-1-methyl-2-cyclohexen-1-ol ((−)-IMCH) as the major component of the aggregation pheromone of P. quercivorus. We tested the ability of synthetic (−)-IMCH to attract P. quercivorus adults. The synthetic compound attracted 14.4 times as many P. quercivorus, of both sexes, as control traps. Control and (−)-IMCH-baited traps both captured 3.32 times as many males as females, probably because of the nature of Lindgren funnel traps and of male-biased emergence sex ratio early in the season of adult flights. (−)-IMCH was proved to be a major component of the aggregation pheromone of P. quercivorus.     

Damage to residual stands from thinning with short-span tower yarders: Re-examination of wounds after five years

The current status of wounds on residual trees was investigated and compared with the wound status from five years ago when the stands were thinned and logged. Wound status was defined according to the four levels of severity based on appearance. Several damaged trees were cut for inspection of the wounded areas. Most wounds of a lighter severity level had already healed by occlusion and were no longer evident. Wounds reflecting heavy damage but had healed, were smaller in the horizontal dimension, or width, than those not yet healed. Slight wounds, but with a current status reflecting persistent damage were smaller in the vertical dimension, or length. An inspection of wound sections revealed the development of discoloration in the wood. Only the slight wounds that healed early had no or small discolored areas. The width of discolored areas of wounds in which cambium was still exposed was greater than the original width of the wound. In conclusion, wound width rather than length significantly affects healing or occlusion of wounds.     

Synthesis and Structure–Activity Relationships of Miticidal 4,5-Dihydropyrazole-5-thiones

A series of novel 4,5-dihydropyrazole-5-thiones (DHPs) was synthesised by treating the corresponding dihydropyrazolones with ‘Lawesson’s reagent and evaluated for miticidal activity against two-spotted spider mites (Tetranychus urticae Koch). Of these, 3-(4-chlorophenyl)-4,4-dimethyl-1-phenyl-4,5-dihydropyrazole-5-thione, 3-(4-chlorophenyl)-4-ethyl-4-methyl-1-phenyl-4,5-dihydropyrazole-5-thione, 3-(4-chlorophenyl)-1-phenyl-4,5-dihydropyrazole-5-thione-4-spirocyclopentane and 4,4-dimethyl-1-phenyl-3-(4-trifluoromethyl-phenyl)-4,5-dihydropyrazole-5-thione were highly active (pEC₅₀>4·0) and were more effective than the miticide dicofol (pEC₅₀=3·879), which has traditionally been used for the control of phytophagous mites. Structure–activity relationship (SAR) studies were performed on each position of the pyrazole ring of DHPs. The results indicated that the unsubstituted phenyl, 4-substituted phenyl and thioxo groups on the 1-, 3- and 5-positions of DHPs respectively were required for activity. Quantitative SAR studies using physicochemical parameters of substituents and the capacity factor k′ as a hydrophobicity index suggested that: (a) the activities of all types of DHPs examined were mainly dominated by hydrophobicity, (b) the bulkiness of 4-substituents of the 3-phenyl ring favoured the activity and (c) the log k′ optimum for all DHPs was 1·675, equivalent to a log P_ow value of c. 5·0.