Duration of calcineurin and Erk signals regulates CD4/CD8 lineage commitment of thymocytes

CD4/CD8 lineage commitment of thymocytes is controlled by the T cell receptor-mediated signals and is mimicked in vitro by a long-pulse stimulation of isolated CD4(+)CD8(+) thymocytes with proper combinations of phorbol myristate acetate and the calcium ionophore ionomycin. CD4 lineage commitment required higher intracellular Ca(2+) levels than CD8 lineage commitment in this culture system. The calcineurin inhibitor FK506 at 1nM inhibited the development of thymocytes to either lineage, but 0.3nM FK506 significantly switched the development from the CD4 cell fate to the CD8 cell fate. The switch in lineage commitment was also observed when 1nM FK506 was added 8h after the start of the culture. Delayed addition of 20microM U0126, an Mek (Erk kinase) inhibitor, also induced the switch. These results suggest that the intensity of calcineurin activity and the duration of both calcineurin and Erk pathway activation are crucial for thymocyte lineage commitment.     

Genetic variants of the receptors for thromboxane A2 and IL-4 in atopic dermatitis

Thromboxane A2 (TXA2) is an arachidonate metabolite which is considered to relate to chronic inflammation in atopic diseases characterized by elevated immunoglobulin E productivity. The elevation of immunoglobulin E levels involves many molecules including interleukin-4 (IL-4) and interleukin-4 receptor alpha chain (IL-4R alpha). To assess whether genetic variants of TXA2 receptor, IL-4 and IL-4R alpha genes relate to the elevation of serum immunoglobulin E levels in patients with atopic dermatitis (AD), we conducted an association study of genetic polymorphisms of TXA2 receptor (795C/T), IL-4 (-589C/T), and IL-4R alpha (Ile50Val) in a Japanese population (n = 789). The TXA2 receptor 795TT genotype strongly related to AD with high serum immunoglobulin E concentrations. AD patients with both TXA2 receptor 795TT genotype and the IL-4R alpha Ile50/Ile50 genotype showed the greatest immunoglobulin E concentrations. These results suggest TXA2 receptor polymorphism strongly interacts with IL-4R alpha polymorphism as a major determinant of high serum immunoglobulin E levels in AD.     

Stretch-activated cation channels in skeletal muscle myotubes from sarcoglycan-deficient hamsters

Deficiency of -sarcoglycan(-SG), a component of the dystrophin-glycoprotein complex, causescardiomyopathy and skeletal muscle dystrophy in Bio14.6 hamsters. Usingcultured myotubes prepared from skeletal muscle of normal and Bio14.6hamsters (J2N-k strain), we investigated the possibility that the-SG deficiency may lead to alterations in ionic conductances, whichmay ultimately lead to myocyte damage. In cell-attached patches (withBa²⁺ as the charge carrier), an ~20-pS channel wasobserved in both control and Bio14.6 myotubes. This channel is alsopermeable to K⁺ and Na⁺ but not toCl. Channel activity was increased by pressure-inducedstretch and was reduced by GdCl₃ (>5 µM). The basal openprobability of this channel was fourfold higher in Bio14.6 myotubes,with longer open and shorter closed times. This was mimicked bydepolymerization of the actin cytoskeleton. In intact Bio14.6 myotubes,the unidirectional basal Ca²⁺ influx was enhanced comparedwith control. This Ca²⁺ influx was sensitive toGdCl₃, signifying that stretch-activated cation channelsmay have been responsible for Ca²⁺ influx in Bio14.6hamster myotubes. These results suggest a possible mechanism by whichcell damage might occur in this animal model of muscular dystrophy.

     

Sequence of IGF-I, IGF-II, and HGF expression in regenerating skeletal muscle

Various cytokines are thought to play a role in muscle regeneration, however, the interaction and mechanisms of action of these cytokines remains largely unknown. In this study, we investigated the role of HGF, IGF-I, and IGF-II during myogenesis using the regeneration model of skeletal muscle as well as myoblast culture. RT-PCR analysis revealed that HGF and IGF-I expressions were markedly upregulated, in regenerating muscle. In contrast, there was no significant difference in IGF-II expression between normal and regenerating muscle. Immunohistochemical analysis demonstrated that HGF was expressed mostly by myocytes during the early stages of muscle regeneration. Additionally, HGF inhibited the formation of myotubes by myoblasts, but promoted cellular proliferation. Otherwise, IGF-I and IGF-II were expressed by myocytes through the early to middle stages of muscle regeneration. The addition of HGF to myoblast growing in vitro significantly increased the number of cells. These findings indicate that these three cytokines have pleiotropic effects in regenerating skeletal muscle.     

Telomerase inhibiting activity in vitro from natural resources, marine algae extracts

We attempted to screen for telomerase inhibitingactivity in vitro from a total of 304 marine algae samples which were collected from various Japancoasts by nonradioisotope telomeric repeatamplification protocol (Non-RI TRAP) assay using humanleukemia MOLT-4 cells. Ten of the MeOH extracts andtwo of the PBS extracts from those algal samplesshowed telomerase inhibiting activity. In particular,the MeOH extract from a green alga, Caulerpasertularioides strongly inhibited telomerase activitywhen added to MOLT-4 cell culture at a level of 1.25%(v/v), suggesting that it may be possible to developa novel anti-cancer agent in view of its specificanti-telomeric property.     

A new species of the genusamblyeleotris (pisces: Gobiidae) from Japan

A new shrimp-associated goby,Amblyeleotris melanocephala, is described on the basis of specimens from Okinoshima Island. Kochi Prefecture, and Okinawa Island, Okinawa Prefecture, Japan. The species is distinguished from other members of the genus by the following combination of characters: head dark brown, a few yellow spots on pectoral fin base and opercular margin, 13 second dorsal and 13 anal fin soft rays, 20 pectoral fin rays, longitudinal scales 92–101, proportional length of interpelvic connecting membrane relative to longest pelvic fin ray (CM-value) 0.46–0.55, presence of a ventral frenum, midline of nape naked, sides scaled above midpoint between preopercle and opercle.     

Whole‐genome prediction of fatty acid composition in meat of Japanese Black cattle

Because fatty acid composition influences the flavor and texture of meat, controlling it is particularly important for cattle breeds such as the Japanese Black, characterized by high meat quality. We evaluated the predictive ability of single‐step genomic best linear unbiased prediction (ssGBLUP) in fatty acid composition of Japanese Black cattle by assessing the composition of seven fatty acids in 3088 cattle, of which 952 had genome‐wide marker genotypes. All sires of the genotyped animals were genotyped, but their dams were not. Cross‐validation was conducted for the 952 animals. The prediction accuracy was higher with ssGBLUP than with best linear unbiased prediction (BLUP) for all traits, and in an empirical investigation, the gain in accuracy of using ssGBLUP over BLUP increased as the deviations in phenotypic values of the animals increased. In addition, the superior accuracy of ssGBLUP tended to be more evident in animals whose maternal grandsire was genotyped than in other animals, although the effect was small.     

The influence of preceding dive cycles on the foraging decisions of Antarctic fur seals

The foraging strategy of many animals is thought to be determined by their past experiences. However, few empirical studies have investigated whether this is true in diving animals. We recorded three-dimensional movements and mouth-opening events from three Antarctic fur seals during their foraging trips to examine how they adapt their behaviour based on past experience—continuing to search for prey in the same area or moving to search in a different place. Each dive cycle was divided into a transit phase and a feeding phase. The linear horizontal distance travelled after feeding phases in each dive was affected by the mouth-opening rate during the previous 244 s, which typically covered two to three dive cycles. The linear distance travelled tended to be shorter when the mouth-opening rate in the previous 244 s was higher, i.e. seals tended to stay in the same areas with high prey-encounter rates. These results indicate that Antarctic fur seals follow decision-making strategies based on the past foraging experience over time periods longer than the immediately preceding dive.     

Establishment and characterization of a human cell line, HS-MM, derived from a case of clear cell sarcoma

The characteristics of a new human clear cell sarcoma (CCSa) cell line, HS-MM, established from the pleural effusion in a 39-year-old man with lung metastasis, have been morphologically studied in vitro and in vivo. HS-MM cells growing on a cover-slip were round or spindle in shape with round nuclei containing extremely prominent nucleoli. Heterotransplantation of the cells into nude mice was easily succeeded following tumor development. Light microscopically, HS-MM cells, both in vitro and in vivo, were positive for anti-S-100 protein and anti-melanoma specific antibodies with immunostain, but no melanin pigment was detected in them. Ultrastructurally, the cells had round euchromatin-rich nuclei with large nucleoli revealing conspicuous nucleolonema, and contained a few mitochondria, rough endoplasmic reticulum and lysosomal dense bodies, besides a large amount of glycogen, but no melanosome in their cytoplasm. HS-MM cells retained and fully expressed morphologically unique characteristics as a CCSa, compatible with amelanotic type of malignant melanoma also. This cell line, HS-MM, therefore, proves to be extremely useful for clinicopathological studies on a CCSa.