Development of a system for measuring calcitonin in the stingray Dasyatis akajei (a cartilaginous fish): the possible involvement of stingray calcitonin in gonadal development

To elucidate the physiological role of calcitonin (CT) in stingrays (cartilaginous fish), an enzyme-linked immunosorbent assay (ELISA) system using a specific antibody against stingray CT has been developed. Synthetic stingray CT was subcutaneously injected into mice four times—once every 2 weeks—together with an adjuvant. We purified the IgG antibody fraction using the protein A affinity chromatography from collected antiserum. Evaluating the antibody titer, we found the antibody’s optimum dilution ratio to be 600 times. Competitive ELISA has been developed using the antibody diluted 600 times. Our antibody did not cross-react with teleost CTs and muscle extraction, but cross-reacted with stingray plasma and the extract of the ultimobranchial gland, the secretary organ of stingray CT. Using this ELISA, we measured the plasma CT level in stingrays and examined its correlation with several mineral concentrations. Plasma CT did not show significant correlation to calcium, magnesium, inorganic phosphorus, sodium, chlorine, or urea, although there was a correlation among the factors involved in osmoregulation, such as sodium, chlorine, and urea. On the other hand, plasma CT was significantly correlated to body weight and length. Furthermore, there was a significant correlation between plasma CT and gonad weight. Since plasma CT was correlated with the weight of liver, which is involved in the synthesis of egg yolk protein, we examined the influence of 17β-estradiol (E2) on CT secretion. After E2 injection, the plasma CT level increased significantly. This is the first study to demonstrate that E2 induced plasma CT secretion in cartilaginous fish.     

Influence of Water Temperature on Morphological Deformities in Cultured Larvae of Japanese Eel,Anguilla japonica,at Completion of Yolk Resorption

The occurrence of morphological deformities under different rearing water temperatures (18, 20, 22, 24, 26, 28, and 30 C) was examined in Japanese eel larvae. The rates of hatching and survival until yolk resorption at 22–26 C were higher than those at other water temperatures. Fertilized eggs never hatched at 18 and 30 C. The rates of occurrence of abnormal larvae reared at the water temperatures 24–28 C were lower than those at 20 or 22 C. Pericardial edema and lower jaw deformities occurred most frequently at lower temperatures (20 and 22 C). In contrast, the incubation temperature did not significantly affect the relative frequency of some neurocranial deformities and of spinal curvature. These results imply that the optimal temperatures for rearing Japanese eel eggs and embryos are 24–26 C from the viewpoints of survival and deformity.     

Changes in free amino acid concentrations in mycorrhizal strawberry plants

The influence of arbuscular mycorrhizal fungi (AMF) colonization on free amino acid concentrations in strawberry (Fragaria × ananassa Duch., cv.Nohime) plants was investigated using two AMF species [Glomus mosseae (Gm), Gl. aggregatum (Ga)] under phosphorus-supplemented (+P) or non-supplemented (−P) conditions. Ten weeks after AMF inoculation, mycorrhizal plants showed higher values in dry weight of both shoots and roots than did non-mycorrhizal ones among most of the treatments. Shoots and roots of mycorrhizal plants had greater phosphorus concentrations in −P plots, while in +P plots, P concentrations differed little among the inoculation treatments. AMF colonization was greater in plants inoculated with Gm than in plants inoculated with Ga. Total amino acid concentration was higher in most of the plants inoculated with Gm than in non-mycorrhizal ones regardless of P treatment. Serine, glutamic acid, glycine, alanine, leucine and GABA were higher in both mycorrhizal plants in −P plots than in non-inoculated plants. In +P plots, threonine and isoleucine concentrations were greater in both mycorrhizal plants than in non-inoculated ones. Asparagine, glutamic acid, glycine, citrulline, GABA and arginine were greater in plants inoculated with Gm than in non-mycorrhizal ones. These findings verified that inoculation with AMF increases total amino acid concentrations and concentrations of specific amino acids in strawberry plants with or without phosphorus supplementation, though the effects varied with species of mycorrhizal fungus.     

Molecular cloning of sex hormone-binding globulin cDNA in carp, Cyprinus carpio

Two types of cDNAs encoding carp SHBGs were isolated and characterized. Northern blot analysis showed their mRNAs distributed mainly in liver, and mRNA levels were almost constant during annual reproductive cycle. This result suggests SHBG synthesis is independent of serum steroid concentration.     

Immunohistochemical studies on ontogeny of bovine lymphoid tissues.

Developing lymphoid tissues of bovine fetuses ranging from 70 to 270 days of fetal age were examined by histological and immunohistochemical procedures. In the peripheral blood, surface membrane immunoglobulin bearing cells (B-lymphocytes) and sheep red blood cell rosette forming cells (T-lymphocytes) had already appeared by 70 days of fetal age. In the lymph nodes intracytoplasmic IgM positive cells appeared at 90 days of fetal age. The cells positive for IgG appeared at 150 days of fetal age and IgA positive cells appeared at 180 days of fetal age. The spleen contained intracytoplasmic immunoglobulin positive cells at almost the same time as those in the lymph nodes. In the ileocecal region, IgM positive cells and IgG positive cells were present at 180 days of fetal age and IgA positive cells were present at 210 days of fetal age. The tonsils contained IgM positive cells and IgG positive cells at 240 days of fetal age. In the thymus, terminal deoxynucleotidyl transferase positive cells appeared at 90 days of fetal age.     

Molecular ordering of organic molten salts triggered by single-walled carbon nanotubes

When mixed with imidazolium ion-based room-temperature ionic liquid, pristine single-walled carbon nanotubes formed gels after being ground. The heavily entangled nanotube bundles were found to untangle within the gel to form much finer bundles. Phase transition and rheological properties suggest that the gels are formed by physical cross-linking of the nanotube bundles, mediated by local molecular ordering of the ionic liquids rather than by entanglement of the nanotubes. The gels were thermally stable and did not shrivel, even under reduced pressure resulting from the nonvolatility of the ionic liquids, but they would readily undergo a gel-to-solid transition on absorbent materials. The use of a polymerizable ionic liquid as the gelling medium allows for the fabrication of a highly electroconductive polymer/nanotube composite material, which showed a substantial enhancement in dynamic hardness.     

Development of a rapid method for identifying asparagus super-males using, N-(4-chloro-2-trifluoromethylphenyl)-N'- propoxyacetamidine to induce flowering

In order to shorten the term for the identification of asparagus super-maleplants, a method usingN-(4-chloro-2-trifluoromethylphenyl)-N'-pro-poxyacetamidine (AM12) toinduce flowering was developed. This method is conducted as follows: seedsfrom andromonoecious flowers are treated with AM12 to induce flowersto form on the seedlings, the new male flowers are crossed with normalfemale flowers, and the progeny seeds are again treated with AM12 toidentify super-male plants from the sex ratio of the seedlings.Asparagus officinalis cv. `UC157' seeds were treated with AM12.The seedlings were induced to flower at a high frequency in 25 days. Thesex ratio of the plants was 1:1 and the male flowers had the pollengermination ability. One male flower induced by AM12 was crossed withfemale plant. This cross produced many progeny seeds. The seeds weretreated again with AM12, and induced to flower. Two super-male plantswere found among the progeny of andromonoecious flowers of the all-malecultivar `Gijnlim' within six months by this method. This method thusshortens the time for identifying super-males. Since the female flowers werefertile as well, AM12 treatment would also be effective for cross breeding.