Nonobese diabetic mouse congenic analysis reveals chromosome 11 locus contributing to diabetes susceptibility, macrophage STAT5 dysfunction, and granulocyte-macrophage colony-stimulating factor overproduction

Unstimulated monocytes of at-risk/type 1 diabetic humans and macrophages of the NOD mouse have markedly elevated autocrine GM-CSF production and persistent STAT5 phosphorylation. We analyzed the relationship between GM-CSF production and persistent STAT5 phosphorylation in NOD macrophages using reciprocal congenic mouse strains containing either diabetes-susceptible NOD (B6.NODC11), or diabetes-resistant C57L (NOD.LC11) loci on chromosome 11. These intervals contain the gene for GM-CSF (Csf2; 53.8 Mb) and those for STAT3, STAT5A, and STAT5B (Stat3, Stat5a, and Stat5b; 100.4-100.6 Mb). High GM-CSF production and persistent STAT5 phosphorylation in unactivated NOD macrophages can be linked to a region (44.9-55.7 Mb) containing the Csf2 gene, but not the Stat3/5a/5b genes. This locus, provisionally called Idd4.3, is upstream of the previously described Idd4.1 and Idd4.2 loci. Idd4.3 encodes an abundance of cytokine genes that use STAT5 in their macrophage activation signaling and contributes approximately 50% of the NOD.LC11 resistance to diabetes.     

Homologies of the adductor mandibulae muscles in Tetraodontiformes as indicated by nerve branching patterns

Homologies of the adductor mandibulae muscles in eight families of Tetraodontiformes were hypothesized from the branching patterns of ramus mandibularis trigeminus. Insertions of the muscles to the upper or lower jaw were weak indicators of homology, migrations of the sites occurring frequently in A1, A2, A2, and A3. In monacanthids, tetraodontids, and diodontids, A1 tended to be split into numerous subsections, whereas in aracanids and ostraciids, A3 was highly developed, comprising three or four subsections. In tetraodontids, A2 was found to be a composite of A1 subsection and A2. The methods of and limits to applying nerve branching patterns to muscle homology are discussed. A new naming system that reflects both muscle homologies and insertions is proposed.     

The occurrence of two species of <Emphasis Type="Italic">Macroramphosus</Emphasis> (Gasterosteiformes: Macroramphosidae) in Japan: morphological and ecological observations on larvae,juveniles, and adults

Earlier opinions that Macroramphosus is monotypic are refuted, with two species apparently occurring in Japan (tentatively identified as M. gracilis and M. scolopax). In postsettlement young and adults, the former is characterized by a dark slender body (vs. red-orange and deep) and short second dorsal fin spine with a smooth posterior margin (vs. long spine with a serrated margin). Food habits also differ between the two species, which are either plankton or benthos feeders. Two types of Macroramphosus larvae and juveniles occurring at the surface were recognized, one having a straight ventral body profile of the body (identified here as M. gracilis) and the other having a notch in the anal region. The dark body of postsettlement M. gracilis is considered to be a retention of the character suited to the neustonic distribution of the larval and juvenile stages, the species remaining to ca. 40mm in standard length (SL) in that habitat (vs. to ca. 12mm SL in M. scolopax).     

Hydrophilic interaction liquid chromatography-tandem mass spectrometry for the determination of levosulpiride in human plasma

A rapid, sensitive and selective hydrophilic interaction liquid chromatography-tandem mass spectrometric (HILIC-MS/MS) method for the determination of levosulpiride in human plasma was developed. Levosulpiride and internal standard, tiapride were extracted from human plasma with ethyl acetate at pH 11 and analyzed on an Atlantis HILIC silica column with the mobile phase of acetonitrile-ammonium formate (190 mM, pH 3.0) (94:6, v/v). The analytes were detected using an electrospray ionization tandem mass spectrometry in the multiple-reaction-monitoring mode. The standard curve was linear (r = 0.999) over the concentration range of 1.00-200 ng/ml. The lower limit of quantification for levosulpiride was 1.00 ng/ml using 100 microl plasma sample. The coefficient of variation and relative error for intra- and inter-assay at three quality control (QC) levels were 3.8-9.1 and -2.9 to -0.1%, respectively. The recoveries of levosulpiride ranged from 80.5 to 87.4%, with that of tiapride (internal standard) being 84.6%. This method was successfully applied to the pharmacokinetic study of levosulpiride in humans.     

Influence of in vitro growth conditions on photosynthetic competence and survival rate of Rehmannia glutinosa plantlets during acclimatization period

The photosynthetic responses of Rehmannia glutinosa grown under photoautotrophic or heterotrophic conditions in vitro were investigated after transfer to greenhouse conditions. In addition, the changes in carbohydrate content and survival rates of the plantlets were evaluated. During six days after transplantation, the photosynthetic rate declined and photoinhibitory impairments represented by decrease of Fv/Fm and chlorophyll content were observed regardless of environmental conditions in vitro. Excessive transpiration was observed in plantlets grown under heterotrophic conditions during that period. Fructose and glucose content of the plantlets grown under photoautotrophic conditions increased with time and reached almost the same level of field grown plants after day 15. Under heterotrophic conditions, in contrast, the content of these sugars decreased continuously during that period. It is suggested that high survival rate of plantlets grown under photoautotrophic conditions has to be attributed to improvement of photosynthetic competence by imposed high light intensity and CO₂ concentration in vitro. The results strongly suggest that the control of transpiration during early stage after transplantation plays a key role in the acclimatization process, and photoautotrophic conditions could be a solution to solve the problems associated with transplantation stress. This revised version was published online in June 2006 with corrections to the Cover Date.     

In-vitro seed germination ofCalanthe sieboldii, an endangered orchid species

Immature seeds from unripe capsules ofCalanthe sieboldii, were sown on one of three sterilized media: MS; modified MSH [i.e., the inorganic salts of MS plus the organic elements of Schenk and Hildebrandt]; Hyponex. Germination and protocorm development occurred on the MS and MSH media within eight weeks, but percent germination was low. The addition of putrescine (1 mg L^-1) or adenine sulfate (25 mg L^-1) to the MSH medium enhanced germination. Resultant protocorms were subcultured on the Hyponex medium, where they developed into plantlets after 12 weeks of additional culture. Plantlets were then successfully transferred to community pots.     

Blue and red light-emitting diodes with or without sucrose and ventilation affect in vitro Growth ofRehmannia glutinosa plantlets

Single-node, in vitro cuttings ofRehmannia glutinosa were transplanted to MS basal media and grown for 30 d. Plantlets were grown under various culture conditions: four different light qualities (red LEDs, blue LEDs, mixed LEDs, and fluorescent); with sucrose (30 mg.L^-1) or without (0 mg.L^-1); with air exchanges (3.5 h.^-1) or without (0.1 h.L^-1). Highest dry weights were obtained from plantlets under blue LEDs with 3.5h.L^-1 air exchanges. Light source did not affect shoot elongation in ventilated conditions, but without ventilation, the shoots of plantlets under red LEDs were twice as long as for plantlets growing under other types of lighting. Plantlets grown without sucrose showed little difference in photosynthesis under any of the tested light qualities. In contrast, the photosynthetic rate of those in the sucrose-containing media varied according to light source.     

Aeration volume and photosynthetic photon flux affect cell growth and secondary metabolite contents in bioreactor cultures ofMorinda citrifolia

To improve their growth and secondary metabolite production, we culturedMorinda citrifolia leaf cells for 3 weeks in bioreactors with different aeration volumes (0.05, 0.1, 0.2, or 0.3 vvm; or 0.05/0.1/0.2/0.3 vvm, as increased at 5-d interval), and photosynthetic photon fluxes (PPF; 0, 15, 30, or 45 μ,moL m^-2 s^-1). Cell growth was greatest (15.6 g L^-1 dry weight) at 0.3 vvm whereas the accumulation of secondary metabolites (total anthraquinones, phenolics, and flavonoids) was maximized at 0.1 vvm. A PPF of 15 μmoLm^-2 s^-1 accelerated the accumulation of both cell biomass and metabolites. Dark-culturing suppressed cell growth, while a high PPF (45 μmoLm^-2 s^-1) inhibited metabolite biosynthesis. Further studies are required to understand the reason for differences in the effect of light on cell growth and secondary metabolite contents inM. citrifolia cell cultures.