Hepatotoxin (microcystin) and neurotoxin (anatoxin-a) contained in natural blooms and strains of cyanobacteria from Japanese freshwaters

Amounts of hepatotoxic microcystin and neurotoxic anatoxin-a were estimated in natural blooms and strains of cyanobacteria from freshwaters in Japan. A simultaneous analysis method of anatoxin-a and microcystin was applied to natural bloom samples, which has been dominated by several species and the strains of cyanobacteria which produced simultaneously both toxins. The natural blooms examined in the present study were mainly composed of Anabaena and Oscillatoria, but most also contained Microcystis and other cyanobacteria. Only one sample was almost unialgal, Anabaena spiroides, collected from Lake Sagami. The toxins in 14 samples collected from nine different natural blooms during 1988-1992 were identified as microcystins-RR, -YR, and -LR; desmethyl-7-microcystin-LR (7-DMLR); and anatoxin-a. Microcystins were the main toxins contained in these natural blooms, with anatoxin-a not being detected or of very little quantity. 7-DMLR was detected in samples only from Lake Kasumigaura. Five strains of Anabaena isolated from waters in Japan produced a small amount of anatoxin-a, but no microcystins. One half of the strains of Microcystis produced microcystins and/or anatoxin-a. This is the first study showing Microcystis producing both anatoxin-a and microcystins.     

F-actin bundling activity of Tetrahymena elongation factor 1 alpha is regulated by Ca2+/calmodulin

Translation elongation factor 1 alpha (EF-1 alpha) catalyzes the GTP-dependent binding of amino-acyl-tRNA to the ribosome. Previously, Tetrahymena 14-nm filament-associated protein was identified as EF-1 alpha [Kurasawa et al. (1992) Exp. Cell Res. 203, 251-258]. This and several other studies suggest that EF-1 alpha functions not only in translation but also in regulation of some part of the cytoskeleton. Tetrahymena EF-1 alpha bound to F-actin and induced bundling of F-actin. We investigated the effects of GTP/GDP and Ca2+/calmodulin on F-actin bundling activity of EF-1alpha. The presence of GTP, GDP, or guanylyl-imidodiphosphate (GMP-PNP) slightly decreased the amount of EF-1 alpha which bound to F-actin, but each had virtually no effect on the F-actin bundling activity. The formation of F-actin bundles by EF-1 alpha was Ca(2+)-insensitive. In the absence of Ca2+, calmodulin did not bind to EF-1 alpha and F-actin. On the other hand, in the presence of Ca2+, calmodulin directly bound to EF-1 alpha but did not have any serious influence on EF-1 alpha/F-actin binding. Under the conditions, electron microscopy demonstrated that Ca2+/calmodulin completely inhibited the F-actin bundling by EF-1 alpha. These results indicate that CA2+/calmodulin regulates the F-actin bundling activity of EF-1 alpha without inhibition of the binding between Ef-1 alpha and F-actin.     

Uniform temperature cooling power of the superfluid Stirling refrigerator

Uniform temperature cooling power measurements of a superfluid Stirling refrigerator are presented for³He-⁴He molar concentrations of 5.9%, 17% and 36% and for temperatures between 0.37 K and 1.4 K. The results are compared to an ideal Fermi gas model and to a more general thermodynamic model. The Fermi model agrees well with the 5.9% concentration data; however, the more elaborate model is needed for higher concentration mixtures.     

Studies on chitin. IX. Crosslinking of water-soluble chitin and evaluation of the products as adsorbents for cupric ion

Water-soluble chitin was successfully crosslinked to varying extents with glutaraldehyde in homogeneous aqueous solutions to improve the properties as an adsorbent for metal cations, and the effects of crosslinking were discussed. Complete insolubilization was achieved with the fivefold excess aldehyde, but, in terms of adsorptivity of Cu²⁺, the chitin crosslinked at an aldehyde/amino group ratio of 1.0 was found to exhibit remarkable capacity and was much superior to others. The desorption of Cu²⁺ from the adsorption complex was also attained effectively at pH 2.0. These results indicated that the loose crosslinking was quite simple and efficient to produce high capacity adsorbents for practical use. Thermal behavior of the crosslinked chitin was examined by TMA and TGA; a softening phenomenon was observed at 145°C.     

Growth of Potassium Titanate Fibers in Flowing N2 Gas

The length of potassium titanate fibers produced by several conventional methods averages 50 μm, with a maximum of 100 μm. Extremely long fibers (most >1000 μm long) were obtained by calcination in N₂ gas flowing at 5.2×10^-4 m/s.     

Analysis of the genes encoding the variable regions of human IgG rheumatoid factor

OBJECTIVE: To better understand the immunoglobulin variable (V) region repertoire of rheumatoid factors (RF). METHODS: We characterized the heavy (H) and light (L) chain gene segments utilized in a monospecific IgG RF secreting hybridoma (AEE111F) which were derived from a patient with rheumatoid arthritis (RA). The hybridoma was established by fusion of a mouse myeloma cell line with bone marrow derived mononuclear cells from a patient with RA. First strand complementary DNA (cDNA) was generated and used for a polymerase chain reaction amplification of the H and L chain V domains. The amplified V domains were sequenced and compared with an extensive database of germline and cDNA V gene segments. RESULTS: The VH sequence was found to be 96% homologous to a previously described fetal VH3 cDNA (60P2). The VL sequence was also highly homologous to the previously described V lambda II gene (96%) derived from a patient with systemic lupus erythematosus which correlated with an 8.12 idiotype (Id), and to an antibacterial antibody against the Haemophilus influenzae type b capsular polysaccharide (94.7%). CONCLUSION: The overlap among this RF VL gene and the 2 reported V lambda sequences of antibodies that expressed anti-DNA related Id and an environmental pathogen specificity suggests that a part of the IgG RF isolated from patients with RA may thus be derived from the physiological natural antibody repertoire during an abnormal immune response and then develop high affinity, monospecific RF by the selection of an antigen driven mechanism.     

Molecular size-dependent leakage of intracellular molecules from frog skeletal muscle fibers permeabilized with beta-escin

The permeability of beta-escin-treated cell membrane was characterized in terms of the permeant molecular size, by monitoring the leak of cytoplasmic molecules in frog skeletal muscle fibers. With a low concentration of beta-escin (5 microM), most of the cellular ATP was lost within 30-40 min (as revealed by rigor force generation), whereas a fluorescence-labeled dextran injected into the cytoplasm (approximately 10 kDa) and cytoplasmic proteins (14-80 kDa) slowly leaked out of the cell. A high concentration of beta-escin (50-100 microM) accelerated the leak of large molecules. Therefore, low concentrations of beta-escin may be used as a means of permeabilizing the cell membrane to relatively small molecules, while retaining a major fraction of the cellular macromolecules.     

Compatibility of alkaline earth metal (Mg, Ca, Sr)-doped lanthanum chromites as separators in planar-type high-temperature solid oxide fuel cells

The mechanical, electrical and thermal properties of alkaline earth metal (Mg, Ca and Sr)-doped LaCrO₃ have been examined as separators in planar-type high-temperature solid oxide fuel cells. The maximum three-point bending strength at 1000°C in air was measured and found to be 186 MPa for LaCr_0.9Mg_0.1O₃, 36 MPa for La_0.9Ca_0.1CrO₃ and 77 MPa for La_0.9Sr_0.1CrO₃. The La_0.8Sr_0.2CrO₃ separator placed in both an oxidizing and a reducing environment at 1000°C showed almost the same electrical conductivities of the H₂ atmosphere, and the conductivity was independent of sample thickness in the range 0.5–3.0 mm. For all the doped LaCrO₃ perovskites, a difference between the thermal expansion behaviours of air and the H₂ atmosphere was observed. In particular, the thermal expansion slope for the first heating cycle under the H₂ atmosphere showed a marked change. The volume changes were due to the formation of oxygen defects in the perovskite structure. This revised version was published online in November 2006 with corrections to the Cover Date.     

Acetazolamide challenge and technetium-99m-ECD versus iodine-123-IMP SPECT in chronic occlusive cerebrovascular disease

We compared the acetazolamide challenge test using 99mTc-ECD SPECT and 123I-IMP SPECT images in patients with chronic occlusive cerebrovascular disease. We also evaluated the usefulness of linearization correction for acetazolamide challenge test of 99mTc-ECD SPECT. METHODS: Twenty patients with unilateral chronic occlusive cerebrovascular disease (10 patients had middle cerebral arterial lesion and 10 had internal carotid lesion) were included in the study. Split-dose (a dose fractioning was 1:2), and sequential SPECT technique was used for 99mTc-ECD SPECT studies while only acetazolamide challenge test studies for 123I-IMP SPECT were performed. Permeability surface area product model (PS model) and back-diffusion model (Lassen's correction) were used for linearization correction of acetazolamide challenge with 99mTc-ECD SPECT. RESULTS: Six of 16 patients with reduced vasodilatory capacity in 123I-IMP SPECT were underestimated by 99mTc-ECD SPECT acetazolamide challenge test. Relative ECD uptake normalized by cerebellar uptake compared with IMP uptake showed a nonlinear relationship, indicating relatively less uptake in high flow range. The underestimations of limited vasodilatory capacity observed in 99mTc-ECD SPECT without linearization correction was modified by linearization algorithm. However, the effect of correction based on PS model was superior than that of Lassen's correction. The corrected 99mTc-ECD uptake ratio, based on PS model, and IMP uptake ratio demonstrated a better linear relationship than that of Lassen's correction. CONCLUSION: Technetium-99m ECD SPECT corrected based on the PS model is a better method of linearization for evaluating cerebrovascular reserve using acetazolamide challenge.     

Measurements ofPVTx properties of refrigerant mixture HFC-32+HFC-125 in the gaseous phase

The experimental 156PVTx properties of an important binary refrigerant mixture, HFC-32 (difluoromethane)+HFC-125 (pentafluorethane), have been measured for three compositions, i.e., 50, 60, and 80 wt% HFC-32, by a constant-mass-method coupled with expansion procedure in an extensive range of temperaturesT from 320 to 440 K, of pressuresP from 1.8 to 5.3 M Pa, and of densities p from 50 to 124 kg · m^–3. The experimental uncertainties of the present measurements are estimated to be within ±7 mK in temperature, ±2 kPa in pressure, ±0.2% in density and ±0.02 wt% of HFC-32. The sample purities are 99.998 wt% for HFC-32 and 99.99 wt% for HFC-125. Seventy-eight second and third virial coeflicients for temperatures from 320 to 440 K have been determined by the present measurements.Paper presented at the Twelfth Symposium on Thermophysical Properties, June 19–24, 1994, Boulder, Colorado, U.S.A.