Alarm pheromone eliciting attack and escape responses in the sugar cane woolly aphid,Ceratovacuna lanigera (Homoptera,Pemphigidae)

All nymphal stages and apterous adults ofCeratovacuna lanigera secreted droplets containing an alarm pheromone from their abdominal cornicles when stimulated with a pin. When fresh droplets on piece of filter paper were put near the aphids, 1st-instar nymphs originating from apterous adults attacked it, but the advanced instar nymphs and adults escaped from the spot. Nymphs and adults originating from alate adults showed escaping behavior only. These different responses strongly correlated with morphological differences, such as relative lengths of the frontal horns and legs. When a syrphid larva,Allograpta javana, was placed on a colony of aphids, many 1st-instar nymphs attacked it using their frontal horns. During the attack, these nymphs spontaneously secreted droplets and adhered them to the syrphid body. When these droplets were placed on the body, the syrphid larva was agressively attacked by additional 1st-instar nymphs.     

Aurora B but Not Rho/MLCK Signaling Is Required for Localization of Diphosphorylated Myosin II Regulatory Light Chain to the Midzone in Cytokinesis

Non-muscle myosin II is stimulated by monophosphorylation of its regulatory light chain (MRLC) at Ser19 (1P-MRLC). MRLC diphosphorylation at Thr18/Ser19 (2P-MRLC) further enhances the ATPase activity of myosin II. Phosphorylated MRLCs localize to the contractile ring and regulate cytokinesis as subunits of activated myosin II. Recently, we reported that 2P-MRLC, but not 1P-MRLC, localizes to the midzone independently of myosin II heavy chain during cytokinesis in cultured mammalian cells. However, the mechanism underlying the distinct localization of 1P- and 2P-MRLC during cytokinesis is unknown. Here, we showed that depletion of the Rho signaling proteins MKLP1, MgcRacGAP, or ECT2 inhibited the localization of 1P-MRLC to the contractile ring but not the localization of 2P-MRLC to the midzone. In contrast, depleting or inhibiting a midzone-localizing kinase, Aurora B, perturbed the localization of 2P-MRLC to the midzone but not the localization of 1P-MRLC to the contractile ring. We did not observe any change in the localization of phosphorylated MRLC in myosin light-chain kinase (MLCK)-inhibited cells. Furrow regression was observed in Aurora B- and 2P-MRLC-inhibited cells but not in 1P-MRLC-perturbed dividing cells. Furthermore, Aurora B bound to 2P-MRLC in vitro and in vivo. These results suggest that Aurora B, but not Rho/MLCK signaling, is essential for the localization of 2P-MRLC to the midzone in dividing HeLa cells.     

Purification, characterization, and studies on biosynthesis of a 59-kDa bone sialic acid-containing protein (BSP) from rat mandible using a monoclonal antibody. Evidence that 59-kDa BSP may be the rat counterpart of human alpha 2-HS glycoprotein and is synthesized by both hepatocytes and osteoblasts.

A monoclonal antibody was raised against a mineralized tissue-specific sialoprotein containing no phosphorus using partially purified noncollagenous bone matrix proteins from rats as antigen. Then the sialoprotein was purified by high performance liquid chromatography from rat mandibulae using the monoclonal antibody as a marker. The sialoprotein (59-kDa bone sialoprotein (BSP)) with a molecular weight of 59,000 contained 1.4% sialic acid but no detectable phosphorus. Immunohistochemical studies with the antibody showed that the protein was specific to mineralized tissues such as bone and dentin, and present in osteoblasts, osteocytes, and bone matrix. No other soft tissues, such as the cartilage, liver, kidney, and periosteum, were stained. However, Western blot analysis showed that plasma contained immunoreactive 59-kDa BSP. The quantitative amino acid composition of 59-kDa BSP resembled that of human alpha 2-HS glycoprotein (alpha 2-HSG) (Lee, C.-C., Bowman, B.H., and Yang, F. (1987) Proc. Natl. Acad. Sci. U.S.A. 84, 4403-4407; Kellermann, J., Haupt, H., Auerswald, E.-A., and Muller-Esterl, W. (1989) J. Biol. Chem. 264, 14121-14128) and rat 64-kDa protein (Franzén, A., and Heineg?rd, D. (1985) in The Chemistry and Biology of Mineralized Tissues (Butler, W.T., ed), p. 132, EBSCO Media, Birmingham, AL). Amino acid sequence analyses of the amino-terminal region and four peptide fragments of 59-kDa BSP revealed that about 50% of the amino acids were homologous with those of human alpha 2-HSG, which is known to be synthesized by the liver, transported in the bloodstream, and incorporated into calcified tissues. But when newborn rat calvaria, primary cultures of osteoblast-rich cells, and adult rat hepatocytes were incubated with radioactive leucine, immunoreactive 59-kDa BSP was detected in their conditioned medium by fluorography. Several characteristics, including the amino acid sequence, suggest that 59-kDa BSP may be the rat counterpart of human alpha 2-HSG. However, rat 59-kDa BSP is a single peptide and synthesized by both osteoblasts and hepatocytes, whereas human alpha 2-HSG is known to be a heterodimer and to be synthesized by the liver.     

Detection of Hydroxyl free Radicals in the Reperfused Primate Heart

Early reperfusion of an ischemic region can result in significant salvage of the area at risk. We show the presence of hydroxyl free radicals at the time of post ischemia reperfusion using electron paramagnetic resonance (EPR) spectroscopy in a macaque model. These free radicals may be formed as a result of reperfusion or may be an un-involved bystander. It is possible that they may be involved in reperfusion injury.     

Target Binding to S100B Reduces Dynamic Properties and Increases Ca2 +-Binding Affinity for Wild Type and EF-Hand Mutant Proteins

Mutations in the second EF-hand (D61N, D63N, D65N, and E72A) of S100B were used to study its Ca^2 + binding and dynamic properties in the absence and presence of a bound target, TRTK-12. With ^D63NS100B as an exception (^D63NK_D = 50 ± 9 μM), Ca^2 + binding to EF2-hand mutants were reduced by more than 8-fold in the absence of TRTK-12 (^D61NK_D = 412 ± 67 μM, ^D65NK_D = 968 ± 171 μM, and ^E72AK_D = 471 ± 133 μM), when compared to wild-type protein (^WTK_D = 56 ± 9 μM). For the TRTK-12 complexes, the Ca^2 +-binding affinity to wild type (^WT + TRTKK_D = 12 ± 10 μM) and the EF2 mutants was increased by 5- to 14-fold versus in the absence of target (^{D61N + TRTK}K_D = 29 ± 1.2 μM, ^{D63N + TRTK}K_D = 10 ± 2.2 μM, ^{D65N + TRTK}K_D = 73 ± 4.4 μM, and ^{E72A + TRTK}K_D = 18 ± 3.7 μM). In addition, R_ex, as measured using relaxation dispersion for side‐chain ¹⁵N resonances of Asn63 (^D63NS100B), was reduced upon TRTK-12 binding when measured by NMR. Likewise, backbone motions on multiple timescales (picoseconds to milliseconds) throughout wild type, ^D61NS100B, ^D63NS100B, and ^D65NS100B were lowered upon binding TRTK-12. However, the X-ray structures of Ca^2 +-bound (2.0 Å) and TRTK-bound (1.2 Å) ^D63NS100B showed no change in Ca^2 + coordination; thus, these and analogous structural data for the wild-type protein could not be used to explain how target binding increased Ca^2 +-binding affinity in solution. Therefore, a model for how S100B–TRTK‐12 complex formation increases Ca^2 + binding is discussed, which considers changes in protein dynamics upon binding the target TRTK-12.     

Long-term hydrochemical monitoring in an Oyasan Experimental Forest Watershed comprised of two small forested watersheds of Japanese cedar and Japanese cypress

Forest ecosystems are self-fertilizing systems, and development of forest stands depends on nutrient supply via biogeochemical cycling within the ecosystem. Therefore, it is important to clarify the nutrient cycle mediating growth and development. In addition, long-term hydrochemical monitoring is needed to understand the influence of environmental changes on biogeochemical cycling in forest ecosystems. The Oyasan Experimental Forest Watershed (OEFW) is located in the Field Museum Oyasan, the university forest of Tokyo University of Agriculture and Technology, in Gunma prefecture, Japan. OEFW comprises two small adjacent forested watersheds—A-watershed and B-watershed—with respective areas of 1.3 and 1.8 ha. A-watershed is a reestablished forest planted with sugi (Japanese cedar; Cryptomeria japonica) and hinoki (Japanese cypress; Chamaecyparis obtusa) in 1976, and has been managed intensively with fertilizer application. By contrast, B-watershed is an established forest planted with sugi and hinoki in 1907. No forest practices have been carried out except for thinning of suppressed trees in 1983. However, the sugi plantation on the lowest slope (18% of the watershed area) was cut in 2000, and sugi was replanted the following year. In this data paper, we present data on the daily precipitation, discharge, pH, and concentrations of major nutrients (Ca²⁺, Mg²⁺, K⁺, Na⁺, NH₄ ⁺, Cl⁻, NO₃ ⁻, and SO₄ ²⁻) in rainwater and stream water since November 1978. The arithmetical mean pH of precipitation, stream water in A- and B-watershed from the beginning of the monitoring to the present were 4.77 ± 0.67, 6.85 ± 0.41 and 6.88 ± 0.36 (average ± SD), respectively. The arithmetical mean concentrations in precipitation in mmol_c L⁻¹ were 0.030 ± 0.030 for Ca²⁺, 0.010 ± 0.011 for Mg²⁺, 0.009 ± 0.013 for K⁺, 0.020 ± 0.024 for Na⁺, 0.035 ± 0.041 for NH₄ ⁺, 0.026 ± 0.029 for Cl⁻, 0.033 ± 0.038 for NO₃ ⁻, and 0.046 ± 0.043 for SO₄ ²⁻. The mean concentrations in stream water in A-watershed were 0.180 ± 0.032 for Ca²⁺, 0.073 ± 0.013 for Mg²⁺, 0.018 ± 0.009 for K⁺, 0.182 ± 0.024 for Na⁺, 0.010 ± 0.010 for NH₄ ⁺, 0.060 ± 0.008 for Cl⁻, 0.111 ± 0.038 for NO₃ ⁻, and 0.074 ± 0.012 for SO₄ ²⁻; whereas for B-watershed the mean concentrations were 0.169 ± 0.025 for Ca²⁺, 0.079 ± 0.016 for Mg²⁺, 0.018 ± 0.005 for K⁺, 0.192 ± 0.026 for Na⁺, 0.010 ± 0.010 for NH₄ ⁺, 0.065 ± 0.010 for Cl⁻, 0.093 ± 0.025 for NO₃ ⁻, and 0.087 ± 0.011 for SO₄ ²⁻.     

Short-Term Dynamics of Tidepool Fish Community: Diel and Seasonal Variation

Short-term temporal dynamics in a tidepool fish community were quantitatively examined through a series of direct observation at consecutive low tides in a temperate-subtropical intertidal of southwestern Japan. The visual observation was sufficiently effective compared with the capture by anaesthetics and allowed a continual, undisturbed monitoring. Species composition markedly varied between consecutive day versus night in spring-autumn, but not in winter. In contrast, species composition on consecutive days and on consecutive nights was relatively stable all through the year. The pattern of diel change in abundance varied among species and seasons. C. gulosus, Istiblennius sp. and Girella spp. showed the tendency of night-time occurrence, while B. fuscus individuals tended to occur by day. These trends of day–night differences were mainly observed in spring and summer and rarely in winter. Size differences between day and night were detected in B. fuscus, C. gulosus, Istiblennius sp. and Girella spp. This tidepool fish community demonstrated variation in abundance and body size on short time scales, which may be attributable to behavioural differences among species.