Cytogenetic analysis and developmental assessment of mouse embryos derived from in vitro fertilization of oocytes reconstructed by meiosis-II chromosome transplantation

An electrofusion methodology for transferring meiosis-II chromosomes (M-II-t) has not been completely established. The present study compared the use of two temperatures (fusion at 37 C for Group A and 25 C for Group B) during an electrofusion procedure for mouse oocyte M-II-t and investigated the cytogenetic normality and developmental competence of embryos derived from in vitro fertilization using oocytes reconstructed by M-II-t. The M-II-t oocytes were fertilized in vitro and cultured to the blastocyst stage; the resultant embryos were analyzed cytogenetically. Subsequently, chromosomal normality of the resultant embryos at the prometaphase stage of first cleavage division and the integrity of the meiosis-II spindles of the reconstructed oocytes were analyzed. The success rate of electrofusion in Group B was 92.1%, which was significantly different from that in Group A (49.2%) (P<0.05). The fertilization rates (A, 80.7%; B, 77.2%) and development rates (A, 70.9%; B, 65.5%) in the M-II-t groups were significantly lower than those in the control group (95.0 and 92.2%, respectively) (P<0.05). The incidence of chromosomal abnormalities in the Group A embryos (20.5%) at the blastocyst stage was significantly higher than that in the control group embryos (8.5%) (P<0.05), but the incidence of chromosomal abnormalities in Group B (12.5%) was not significantly different compared with the other groups. A temperature of 25 C during the electrofusion procedure for M-II-t resulted in a good fusion rate, good development rate, and efficient production of chromosomally normal blastocysts. Furthermore, the incidence of chromosomal abnormalities in the first cleavage embryos at the prometaphase stage in Group B (9.6%) did not differ significantly from that in the control group (6.6%). The spindle morphology of the M-II-t oocytes in Group B was normal.     

Effect of ovarian hormones on maturation of dendritic cells from peripheral blood monocytes in dogs

Previously, we reported that ovarian hormones affect the immune response against E. coli isolated from the dogs affected with pyometra. In order to investigate mechanisms underlying the immune modulation, we examined the effects of ovarian hormones on the generation of dendritic cells (DCs), the most potent antigen presenting cell. DCs were differentiated from peripheral blood monocytes (PBMOs) using a cytokine cocktail. Both estrogen receptor and progesterone receptors were expressed by the PBMOs and immature DCs. When various ovarian hormones were added to the culture for the DC differentiation, progesterone significantly decreased the expression of DC maturation markers, such as CD1a, CD80 and CD86, on mature DCs. Conversely, the addition of estrogen to the cultures increased the expression of CD86, but not other maturation makers. Furthermore, DCs differentiated in the presence of progesterone did not stimulate allogeneic mononuclear cells in PB. Taken together, these results indicate that progesterone diminishes the maturation of DCs, leading to decreased immune responses against invading pathogens.     

Effects of Calcium Lactate on the Development of Chicken Embryos in a Shell-less Culture System up to Day Seventeen of Incubation

This study examined the effects of calcium lactate on the development of chicken embryos in a shell-less culture system (cSLCS) up to the seventeenth day of incubation. In the presence of calcium lactate, a significant reduction in embryo viability was observed during the first week of incubation in cSLCS. On day 17 of embryo development, no significant difference was observed in the blood plasma calcium concentration or tibia bone density between cSLCS and intact control embryos, whereas the tibia length was significantly shorter in cSLCS embryos than in the intact control. These results suggest that calcium lactate supplementation in cSLCS supports bone formation in developing chicken embryos, but has adverse effects on the viability of embryos, particularly during the first week of embryo development.     

Investigations on the radioactive contamination of crop plants as a result of H-bomb detonation (Part 2) Root and foliage uptake of “Bikini ash”

The Foliar Uptake by Squash Plant

The radioactive ash for experimental use, hereafter referred to as “Bikini ash”, was prepared by igniting the heavily contaminated substances on board No. 5 Fukuryu Maru at about 650°C, followed by sifting through a 100 mesh sieve. On ignition some parts of the fission products, particularly iodine, ruthenium and tellurium would have possibly been lost to the air.     

Distribution of mecA-harboring staphylococci in healthy mares

The prevalence of staphylococci that harbor the mecA gene responsible for methicillin resistance was examined in healthy breeding mares. Staphylococci often cause diseases of horses such as metritis, keratitis, and abscess. Methicillin-resistant staphylococci would make antibiotic treatments ineffective, so it may be significant to know the distribution of mecA-harboring staphylococci in mares. Isolation of mecA-harboring staphylococci was achieved from nares and pasterns of 100 mares in Hokkaido, Japan. From 13% of the mares, mecA-harboring staphylococci, including 15 isolates of Staphylococcus sciuri and 3 of Staphylococcus lentus, were isolated. Isolates of S. sciuri were found to be genetically polyclonal by pulsed-field gel electrophoresis. These isolates produced no PCase and showed low or no resistance to beta-lactam and other classes of antibiotics. Distribution of staphylococcal species and levels of antibiotic resistance were found to be different between isolates from the present mares and those previously reported from riding-horses. Antibiotic pressure may lead to these differences. In addition, it appears that mecA-harboring S. sciuri may be native to horses.     

Simple method for synthesizing phenolicβ-O-4 dilignols

A modified synthetic method for phenolic-O-4 lignin substructure model dimers was developed involving protection of the phenolic hydroxyl group of acetophenons with benzoyl chloride, bromination with 4-dimethylaminopyridiniumbromide perbromide, condensation with phenols in the presence of 18-crown-6-ether, condensation with paraformaldehyde, reduction with NaBH₄ and debenzoylation. This method results in shorter reaction times and increasing yields without the application of strict anhydrous and drastic conditions or chloric solvents. This alternative route could be applied to the-O-4 dilignol syntheses of four combinations of guaiacyl and syringyl derivatives.     

The functional effect of Gly209 and Ile213 substitutions on lysozyme activity of family 19 chitinase encoded by cyanophage Ma-LMM01

ORF69 in the cyanophage infecting Microcystis aeruginosa, Ma-LMM01, shows homology to the family 19 chitinases where the catalytic domain has structural similarity to lysozyme. Chitinases hydrolyze chitin, a β-1, 4-linked monopolymer of N-acetylglucosamine (GlcNAc); whereas lysozymes hydrolyzes peptidoglycan, alternating β-1, 4-linked copolymers of N-acetylmuramic acid (MurNAc) and GlcNAc. Using amino acid sequence comparison to ORF69, the putative sugar binding residues, Gln162 and Lys165, from the barley chitinase (the model enzyme for the family 19 chitinases) corresponding to subsites −4 and −3 were found to be replaced with Gly209 and Ile213, respectively, in ORF69. To analyze their contribution to substrate binding affinity, ORF69 was cloned into Escherichia coli; and two mutant proteins G209Q and I213K were prepared using site-directed mutagenesis. The wild-type gene product (gp69) showed both lysozyme and chitinase activities. In contrast, the I213K mutant showed a decrease (70%) in lysozyme activity and a significant increase (50%) in chitinase activity; whereas, the G209Q mutant almost completely abolished both enzyme activities. The data suggest the Ile213 residue is involved in recognizing the substrate MurNAc; and Gly209 has significant contribution in chitinase and lysozyme activities for the wild-type gp69.     

Sorption of Cr(VI) on the wood of Japanese larch treated with concentrated sulfuric acid

A carbonaceous sorbent was prepared from the wood of Japanese larch (Larix leptolepis) by dehydration with concentrated sulfuric acid in a 69% yield. The abilities of the sorbent to remove Cr(VI) from aqueous solutions were investigated. Research parameters included the initial solution pH, temperature, and initial concentration of Cr(VI) in solution. The removal of Cr(VI) was highly solution pH dependent and was mainly governed by physicochemical sorption under weak acidic conditions. The equilibrium data fit well in the Langmuir isotherm model. The Langmuir constants were calculated at different temperatures, and the sorption capacity increased with rising temperature, indicating the endothermic nature of the Cr(VI) sorption onto the sorbent. The desorption experiments suggest that the Cr(VI) sorption is generally irreversible, owing to strong interaction of HCrO ₄ ⁻ with the active sites of the sorbent.     

Diving behavior of immature, feeding Pacific bluefin tuna (Thunnus thynnus orientalis) in relation to season and area: the East China Sea and the Kuroshio–Oyashio transition region

Twenty‐four archival tags were recovered from Pacific bluefin tuna previously released off Tsushima Island in the East China Sea. By analysis of the time‐series data of the pressure and the ambient and internal temperature from the 24 tags, we examined the relationship between the tuna's pattern of diving and the thermocline depth. In the East China Sea, diving and feeding events occurred throughout almost the entire day in both winter and summer, suggesting that the purpose of diving is for feeding. In summer, the feeding frequency was greater than that in winter, which corresponds to the fact that growth is more rapid in summer than in winter. During summer in the Kuroshio–Oyashio transition region, on the other hand, feeding events were much more frequent than those in the East China Sea, in spite of a lower diving frequency. The mean horizontal distance traveled was also significantly higher and it seems that in this area they may move horizontally to feed on prey accumulated at the surface. We conclude that, in addition to the ambient temperature structure, the vertical and horizontal distribution of prey species plays an important role in the feeding behavior of Pacific bluefin tuna. One bluefin tuna migrated to the Oyashio frontal area, where both the horizontal and the vertical thermal gradients are much steeper. The fish spent most of the time on the warmer side of the front and often traveled horizontally to the colder side during the day, perhaps to feed. This implies that there is a thermal barrier effect, in this case from the Oyashio front, on their behavior. The frequency of feeding events was low, although all the monitored fish dived every dawn and dusk, irrespective of the seasons or location. It is possible that these twice‐daily diving patterns occurred in response to the change in ambient light at sunrise and sunset.