Induction of multiple embryos with NaHCO3 or calcium free sea water in the horseshoe crab

Summary When horseshoe crab embryos were treated with NaHCO₃ at the developmental stage when the germ disc appears, multiple embryos were formed. NaHCO₃ may effect the formation of multiple embryos by binding Ca²⁺ ions of the embryo since multiple embryos were also formed by treatment with Ca²⁺ free sea water.The treatment caused the blastoderm layer to tear. When the embryos were returned to normal sea water after the treatment, the blastoderm recovered. Some cell masses, probably derived from the germ disc or its prospective cells, formed during the process of the recovery. Each cell mass developed into an embryo.Contributions from the Shimoda Marine Research Center, Nor. 348     

Veneza zonata (Hemiptera: Coreidae)/trypanosomatid relationship: action of hemolymph in vitro and experimental infection

The defense response of Veneza zonata (Hemiptera: Coreidae) against three different trypanosomatid infections was assessed: (1) strain 714TD, a Leptomonas which has V. zonata as vector of a plant trypanosomatid, (2) strain 563TD, a Leptomonas isolated from the digestive tract of Euchistus heros (Hemiptera: Pentatomidae), and (3) Leishmania (L.) amazonensis, a human parasite that cannot infect V. zonata. Experiments with V. zonata hemolymph showed agglutination only of L. (L.) amazonensis culture forms and hemocytic recognition was more intense with this strain. L. (L.) amazonensis also activated the prophenoloxidase system, whereas strains 714TD and 563TD did not activate this system but rather seemed to inhibit phenoloxidase activity. No flagellates were seen in the digestive tract, hemolymph, or salivary glands in insects infected with L. (L.) amazonensis. The digestive tract, the hemolymph, and the salivary glands of insects fed on tomatoes inoculated with 714TD are sequentially invaded by the flagellate, which is inoculated in plants together with saliva. Insects fed on tomatoes inoculated with 563TD exhibited culture forms in the digestive tract (6 days after) and hemocoel (three additional days); however, they died 12 to 14 days after exposure. The salivary glands in insects inoculated in the hemocoel with 714TD strain are rapidly invaded, whereas those with 563TD culture forms died approximately 24 h after infection. Bacterial proliferation in the hemocoel and hemocyte surface blebbing were seen in insects infected only with 563TD strain as the probable pathogenic mechanism of insect death.     

Determination of polycyclic aromatic hydrocarbons in milk samples by high-performance liquid chromatography with fluorescence detection

This paper describes a high-performance liquid chromatographic (HPLC) method for the determination of polycyclic aromatic hydrocarbons (PAHs) in milk samples. The method involves a liquid-liquid extraction procedure after saponification of milk samples with sodium hydroxide. Reproducible determination with highly sensitive detection was attained by HPLC with fluorescence detection using 1,2-bis(9-anthryl)ethane as an internal standard. The detection limits of 12 kinds of PAHs ranged from 1.3 to 76 ng/kg milk at a signal/noise ratio of 3. By the proposed method, the presence of 12 and 11 kinds of PAHs could be confirmed in commercial milk and human milk samples, respectively. The average concentrations of total PAHs (mean+/-SD, micro g/kg) were found to be 0.99+/-0.37 for commercial milk (n=14), 2.01+/-0.30 for infant formula (n=3) and 0.75+/-0.47 for human milk (n=51). High correlation coefficients between the concentrations of total PAHs and triglyceride were observed for commercial milk (r=0.659) and human milk (r=0.645).     

Stimulation of Locomotion of Peripheral Blood Monocytes by Human Plasma Fibronectin

The motility of human peripheral blood granulocytes and monocytes in response to human plasma fibronectin was quantified by an in vitro assay using blind-well chemotaxis chambers. Purified fibronectin under nondenaturing conditions produced increased migration of granulocytes only at concentrations higher than 100 nm, and induced increased chemotactic and random locomotion of monocytes at concentrations higher than 0.1 nm. The monocyte migration-inducing activity of fibronectin was concentration dependent, and was strongly inhibited by low concentrations of colchicine (100 nm–100 μm). These findings suggest the possibility that plasma fibronectin serves as a chemotactic stimulus for monocytes in vivo and attracts these cells to sites of microscopic tissue injury where plasma fibronectin is deposited.     

Requirements of the RNA polymerase II C-terminal domain for reconstituting pre-mRNA 3' cleavage

RNA polymerase II (RNAP II) has previously been shown to be required for the pre-mRNA polyadenylation cleavage reaction in vitro. This activity was found to reside solely in the C-terminal domain (CTD) of the enzyme's largest subunit. Using a deletion analysis of glutathione S-transferase-CTD fusion proteins, we searched among the CTD's 52 imperfectly repetitive heptapeptides for the minimal subset that possesses this property. We found that heptads in the vicinity of 30 to 37 contribute modestly more than other sections, but that no specific subsection of the CTD is necessary or sufficient for cleavage. To investigate further the heptad requirements for cleavage, we constructed a series of all-consensus CTDs having 13, 26, 39, and 52 YSPTSPS repeats. We found that the nonconsensus CTD heptads are together responsible for only 20% of the wild-type cleavage activity. Analysis of the all-consensus CTD series revealed that the remaining 80% of the CTD-dependent cleavage activity directly correlates with CTD length, with significant activity requiring approximately 26 or more repeats. These results are consistent with a scaffolding role for the RNAP II CTD in the pre-mRNA cleavage reaction.     

Aboveground biomass and soil nutrient pools of a Scalesia pedunculata montane forest on Santa Cruz, Galápagos

We estimated the live aboveground biomass (AGB) and soil nutrient pools of the Scalesia pedunculata monodominant tropical montane forest at 600 m above sea level on Santa Cruz, Galápagos, an isolated oceanic island. The estimated AGB was 60.4 Mg ha^–1, which was considerably lower than that of other montane forests of similar climates elsewhere. Nutrient pools were ample for inorganic N, soluble P, and exchangeable cations. We suggest that the low AGB, in spite of the ample nutrients, is related to the absence of tall-statured climax species, that have high demands for nutrients (particularly N) to fix C, due to the isolation.     

Species turnover and diversity patterns along an evergreen broad-leaved forest coenocline

Direct gradient analysis was applied to the evergreen broad-leaved forest coenocline in the Tatera Forest Reserve, Japan. 10 quadrats of 0.1 -0.05 ha were laid out from 140 m to 560 m above sea level at intervals of 25–70 m. Gradient analysis revealed that distributions of many species terminated or started at ca. 400 m. Community similarity, calculated in Percentage Similarity (PS) and Community Coefficient (CC), changed abruptly below and above the 400 m contour, suggesting a change of vegetation structure at this altitude, which was also clear from population distributions. The spatial turnover rate of species along the altitudinal gradient was calculated in two ways: as the Average turnover rate along the whole range of the gradient, and as the Zone turnover rate at individual altitudes. The overall rates calculated for five categories of populations: DBH > 10 cm, DBH >3 cm, all woody species, herb-layer, and total vegetation, were- 0.0011 to- 0.0021 for PS, and - 0.0009 to- 0.0019 for CC. The calculated rates (PS basis) indicate that a 95% change in species composition is reached at 1120 to 620 m altitude. Similarly, the rates -0.0009 to - 0.0019 (CC) correspond to 1410 - 680 m. The altitudinal range expected here for a 95% change agrees with the actual elevation of forest zonation in northwestern Kyushu. The average rate of both PS and CC in the herb-layer population was 1.56 times higher than the rate in the woody species population, showing a more rapid change in herb-layer population than in the woody ones along the gradient. The Zone turnover rates were higher at the 370–440 m belt than those below and above the belt. This coincided with the interchanging pattern in population distributions and the abrupt change in similarity at about 400 m above sea level. This may be due to the change in environmental conditions such as physiography and air humidity. In the diversity measurements, the species density per 100 m² showed a gradual increase in the DBH >3 cm population but a constant level in the DBH >10 cm population along the whole range of the forest coenocline studied, while index values of S₍₁₀₀₎ and Shannon's H showed decreasing trends in the same gradient with a few exceptionally high and low values.     

Species diversity of mainland-and island forests in the Pacific area

Alpha diversity, or species richness, of East Asian mainland evergreen broadleaved forests, expressed by indices of Fisher's alpha () and S(₁₀₀), a new index showing species number in a 100-individual sample, is significantly correlated with the climatic favorableness, expressed by Kira's warmth index. On the contrary, diversity values of insular forests studied on Kyushu satellites of Japan, the Bonins, the Eastern Carolines of Micronesia, and the Galápagos in the eastern Pacific, are below those expected from the climate of respective oceanic islands. Species-individual curves, comparing mainland-and insular communities, also support clearly the above conclusion of species poverty in the insular communities studied.Abbreviations WI Kira's (1977) warmth index.