Reproductive ecology and adaptive host choice correlated with body size in an autumn-spawning bitterling Acheilognathus typus.

The life history and reproductive ecology of an autumn-spawning bitterling Acheilognathus typus were studied under natural and experimental conditions. In the study pond, the embryos of A. typus emerged from mussels in May and grew rapidly until August, whereas overwintered age-1 fish grew slowly. Adult A. typus in the pond was smaller (32-47 mm in standard length) than they were in other habitats and mainly spawned in smaller mussels. The number of A. typus embryos in mussels was negatively correlated with the shell length of the mussel, and a lower number of embryos were observed in larger mussels (over 110 mm in shell length). In the mussel size-choice experiment conducted in an enclosure, smaller A. typus selected smaller mussels, and larger A. typus selected larger mussels for spawning. In some cases, smaller A. typus spawned in larger mussels and the number of spawned eggs ejected increased by over four times compared with other cases. These results of the enclosure experiment explained the lower number of embryos in larger mussels in the study pond. In addition, reproductive traits such as ovipositor length and the number of ovulated eggs of female A. typus, which are considered to contribute to their size-dependent host utilization, were positively correlated with their standard length. Because A. typus is geologically or seasonally isolated from other bitterling species, this size-dependent host utilization contributes to a reduction in intraspecies rather than interspecies competition.

Mitochondrial genotyping of an endangered bitterling Acheilognathus typus (Cyprinidae).

Genotyping of endangered species is helpful for establishing and evaluating conservation strategies. Mitochondrial sequence data was analyzed from 541 individuals of a critically endangered fish, Acheilognathus typus from present-day range-wide localities to re-evaluate an in-progress restoration program around Lake Izunuma-Uchinuma, Miyagi, Japan. Acheilognathus typus showed low sequence diversity with only eight haplotypes and π and h values of 0.59129 and 0.00118 respectively. Genetic data suggests Acheilognathus typus is adapted to pulsed environments and prone to population flush and crash. Genotyping of populations in introduced localities revealed that their source is not from nearby localities.

The influence of orifice sealing with various filling materials on coronal leakage.

The aim of this study was to evaluate the sealing ability of materials filled in the orifice after root canal treatment. A total of 100 root canal-treated teeth were divided into six experimental groups: 1, Protect Liner F (PL); 2, Panavia F (PF); 3, DC core-Light cured (DCL); 4, DC core-Chemically cured (DCC); 5, Super-EBA (SE); 6, Ketac (KC). The materials were filled--to a depth of 4 mm--in the coronal part of the root canals, and evaluated for microleakage. The number of teeth that failed to stop dye penetration in the filled materials differed statistically between PL and DCL or SE or KC, PF and SE or KC, DCC and KC, DCL and KC. The mean distance of dye penetration differed significantly between PL and SE or DCC, PF and SE or DCC. Hence, these results indicated the advantageous sealing ability of adhesive and flowable materials.

Molecular and cell biological properties of mouse osteogenic mesenchymal progenitor cells, Kusa.

A cell line of murine osteogenic progenitor cells, Kusa, was established from femoral bone marrow stromal cells with other types of mesenchymal progenitor cells. We characterized two sublines of Kusa (Kusa-A1 and Kusa-O) from several aspects, including the use of an expression profiling system, a cDNA microarray. The original Kusa subline (Kusa-A1) had high alkaline phosphatase activity and high accumulation of calcium deposits in a condition inducing mineralization, with ascorbic acid and beta-glycerophosphate. Kusa-O, a low osteogenic subline of Kusa, had high alkaline phosphatase activity but slow accumulation of calcium deposits even in the inducing condition. These two Kusa sublines differed in the expression of the osteogenic marker genes, osteocalcin and osteopontin, during mineralization. A type of cDNA microarray revealed marked downregulation of gene expression in the inducing condition in both Kusa-A1 and Kusa-O. Another type of high-throughput microarray was performed to examine the difference in gene expression patterns between Kusa-A1 and Kusa-O. By this analysis, periostin, which would be involved in a stage of osteogenesis, was low in Kusa-A1. On the contrary, Myocyte enhancer factor 2C (MEF2C), a myogenic transcriptional factor, was high in Kusa-A1, although no expression of any other myogenic genes was shown.

Osteogenic differentiation of the mesenchymal progenitor cells, Kusa is suppressed by Notch signaling.

Notch receptor plays a crucial role in proliferation and differentiation of many cell types. To elucidate the function of Notch signaling in osteogenesis, we transfected the constitutively active Notch1 (Notch intracellular domain, NICD) into two different osteoblastic mesenchymal cell lines, KusaA and KusaO, and examined the changes of their osteogenic potentials. In NICD stable transformants (KusaA(NICD) and KusaO(NICD)), osteogenic properties including alkaline phosphatase activity, expression of osteocalcin and type I collagen, and in vitro calcification were suppressed. Transient transfection of NICD attenuated the promoter activities of Cbfa1 and Ose2 element. KusaA was capable of forming trabecular bone-like tissues when injected into mouse abdomen, but this in vivo bone forming activity was significantly suppressed in KusaA(NICD). Osteoclasts were induced in the KusaA-derived bone-like tissues, but lacked in the KusaA(NICD)-derived tissues. These results suggest that Notch signaling suppresses the osteoblastic differentiation of mesenchymal progenitor cells.