Identification of ommastrephid squid paralarvae collected in northern Hawaiian waters and phylogenetic implications for the family Ommastrephidae using mtDNA analysis

ABSTRACT:   A total of 110 adult individuals from four ommastrephid (family Ommastrephidae) squid species ( Ommastrephes bartramii, Sthenoteuthis oualaniensis, Eucleoteuthis luminosa, and Hyaloteuthis pelagica ) were used to obtain diagnostic DNA markers for species identification. Restriction fragment length polymorphism (RFLP) analysis of a partial segment (855 bp) of the mitochondrial DNA cytochrome oxidase I (COI) gene amplified by polymerase chain reaction (PCR) revealed that the restriction profiles of two endonucleases ( Alu  I and Tsp 509 I) were diagnostic for species identification. The restriction assay partially supplemented with nucleotide sequence analysis successfully assigned 69 damaged and morphologically equivocal ommastrephid paralarvae collected in northern Hawaiian waters, identifying 60 O. bartramii , eight S. oualaniensis , and one E. luminosa . The family Ommastrephidae appears to be monophyletic. Although the phylogenetic relationships among genera were not resolved well due to apparent homoplasy and large genetic divergence between species, COI sequence data without transitions provided support for subfamily level relationships.     

Peripartum back fat thickness of multiparous Holstein-Friesian cows with displacement of the abomasum or ketosis

To establish a method to predict postpartum diseases using prepartum back fat thickness (BFT), the peripartum BFTs of 54 healthy multiparous cows before calving, which were diagnosed with postpartum displacement of the abomasum (DA), clinical ketosis or subclinical ketosis were compared with those of healthy cows from 8 weeks before the expected calving date to 8 weeks after calving. The peripartum BFTs of the cows with DA or clinical ketosis were significantly higher than those of healthy cows. The peripartum BFTs of the cows with subclinical ketosis were not significantly higher than those of the healthy cows.     

Current classification of Ralstonia solanacearum and genetic diversity of the strains in Japan

Ralstonia solanacearum is the causal organism of bacterial wilt of more than 200 species representing 50 families of plants in tropical, subtropical, and warm temperate regions in the world. Traditionally classified into five races based on differences in host range, R. solanacearum has also been grouped into six biovars on the basis of biochemical properties. With recent developments in molecular biology, various DNA-based analyses have been introduced and used to confirm that this binary system does not completely represent the diversity within R. solanacearum strains. Therefore, a new hierarchical classification scheme has been suggested, which defines R. solanacearum as a species complex and reorganized the concept of the species as a monophyletic cluster according to a phylogenetic analysis based on genomic sequence data. Here we discuss the current bacterial wilt situation and genetic relationships based on the recent classification system of Japanese R. solanacearum strains as well as worldwide strains. We also review the genetic, biochemical, and pathological characteristics of R. solanacearum strains, in particular, those affecting potato and Zingiberaceae plants as distinctly important pathogens in relation to continuously problematic and recent emergent diseases in Japan.     

Primer-directed enzymatic amplification of DNA with a thermostable DNA polymerase

A thermostable DNA polymerase was used in an in vitro DNA amplification procedure, the polymerase chain reaction. The enzyme, isolated from Thermus aquaticus, greatly simplifies the procedure and, by enabling the amplification reaction to be performed at higher temperatures, significantly improves the specificity, yield, sensitivity, and length of products that can be amplified. Single-copy genomic sequences were amplified by a factor of more than 10 million with very high specificity, and DNA segments up to 2000 base pairs were readily amplified. In addition, the method was used to amplify and detect a target DNA molecule present only once in a sample of 10(5) cells.     

Perceptual learning incepted by decoded fMRI neurofeedback without stimulus presentation

It is controversial whether the adult primate early visual cortex is sufficiently plastic to cause visual perceptual learning (VPL). The controversy occurs partially because most VPL studies have examined correlations between behavioral and neural activity changes rather than cause-and-effect relationships. With an online-feedback method that uses decoded functional magnetic resonance imaging (fMRI) signals, we induced activity patterns only in early visual cortex corresponding to an orientation without stimulus presentation or participants' awareness of what was to be learned. The induced activation caused VPL specific to the orientation. These results suggest that early visual areas are so plastic that mere inductions of activity patterns are sufficient to cause VPL. This technique can induce plasticity in a highly selective manner, potentially leading to powerful training and rehabilitative protocols.     

The amount of heavy metals derived from domestic sources in Japan

The amount of heavy metals discharged from daily life was estimated in order to know the origin of the metals contained in the sewage sludge which is produced in the wastewater treatment plant treating only domestic wastewater. The amounts of the metals discharged from our daily life were estimated to be in the range of 0.2 to 0.3 for Cd, 1.6 to 1.9 for Ni, 3.5 to 6.8 for Pb, 0.8 to 1.4 for Cr, 8.2 to 19.3 for Mn 9.4 to 55.8 for Cu, 44.3 to 62.7 for Zn and 111 to 293 for Fe in mg.d^–1 per person, Using these data, the cycle of the metals in our daily life was discussed in relation to land application of sewage sludge.     

Quantities and types of ceramides and their relationships to physical properties of the horn covering the claws of clinically normal cows and cows with subclinical laminitis

Quantities and types of ceramides and their relationships to physical properties of the horn covering the claws of clinically normal cows and cows with subclinical laminitis were investigated. Total ceramide content of the horn covering the sole and wall from cows with subclinical laminitis was 872.2 +/- 146.6 microg/g and 528.6 +/- 61.3 microg/g, respectively, and was significantly (P < 0.01, 0.05) lower than that from clinically normal cows. The mean moisture content in the claws from cows with subclinical laminitis (43.5% +/- 4.3%) was significantly (P < 0.05) higher than that in the claws from clinically normal cows. The hardness of claws from cows with subclinical laminitis (35.2 +/- 3.5) was significantly (P < 0.05) less than that of claws from clinically normal cows. Significant correlations between ceramides and moisture content (P < 0.001) and between ceramide and hardness (P < 0.001) were found in clinically normal cows and cows with subclinical laminitis. Our results indicate that decreases in ceramide contents may be related to changes in physical properties of the horn covering the claw in cows with subclinical laminitis.     

Time-dependent changes in protein phosphorylation patterns in rat brain synaptosomes caused by deltamethrin

Effects of deltamethrin, a powerful pyrethroid insecticide, on the protein phosphorylation and dephosphorylation processes during depolarization in rat brain synaptosomes were studied by using [³²P]phosphoric acid as a starting radiotracer and high external concentration of potassium ions or veratridine (10^?-5 M) as depolarizing agents. At the onset of depolarization there was a quick rise in phosphorylation in various synaptic proteins for about 15–30 s followed by a gradual decline in levels of phosphorylation. The effect of deltamethrin (10^?-7 M) on this system was found to be dependent on the length of preincubation of the synaptosome with the pesticide prior to depolarization. At an early stage (0–3 min preincubation period) it caused a modest suppression of protein phosphorylation activities. When the period of deltamethrin preincubation was extended to 5–20 min, however, it caused a significant increase in protein phosphorylation throughout the depolarization period. At the later stage of the action of deltamethrin (e.g. preincubation period of 30–40 min), deltamethrin-treated synaptosomes no longer responded to the depolarization signal to raise the level of phosphorylation on many proteins. These results indicate that deltamethrin's actions on the synaptic process are complex. Depending on the length of exposure, its effects on protein phosphorylation responses in intact synaptosomes could be either stimulatory or inhibitory. To study the cause of deltamethrin-induced synaptic block at the later stage, effects of deltamethrin on protein kinases were studied by using lysed synaptic membranes with [gamma-³²P]ATP. Deltamethrin was shown to inhibit calcium–calmodulin-dependent protein phosphorylation activities at 10^?-7 M when given directly to the enzyme source 10 min prior to the addition of [³²P]ATP. Such an observation helps to explain the inhibitory action of deltamethrin on protein phosphorylation which occurs at the late stage of its action (i.e. preincubation time > 20 min).     

Progesterone receptor mRNA levels during pregnancy, labor, lactation and the estrous cycle in rat uterus

Progesterone plays important roles in the regulation of female reproduction. In this study, progesterone receptor (PR) mRNA levels in rat uterus during pregnancy, labor, lactation and the estrous cycle were examined by competitive RT-PCR. During pregnancy and lactation, PR mRNA levels had decreased on day 20 of pregnancy (P20) and P21 compared with P15 but increased during labor. After a decline on day 1 of lactation (L1), PR mRNA levels had increased again on L3 and L14 compared with P15, P18, P20, P21 and P21pm (at 2200-2300 h on P21). There was no significant change in the PR mRNA level during the estrous cycle. The PR mRNA level did not change during 1 week of progesterone treatment or afterwards. Injection of 17beta-estradiol did not affect PR mRNA levels in rats treated with progesterone or those without any injections. In rats on P18, 17beta-estradiol injection did not change PR mRNA levels after sham-operation but induced an increase in PR mRNA levels of rats ovariectomized 6 h before the treatment. These results suggest that uterine PR mRNA levels are differently regulated during late pregnancy, labor and lactation, and during labor estrogen is one of the essential factors for the increase in PR mRNA levels.