Ecological and morphological characteristics of the sclerotia of Rhizoctonia solani Kühn produced in soil

Ecological and morphological characteristics of the sclerotia of R. solani produced in soil were compared among Japanese isolates belonging to five anastomosis groups (AG). Twenty-three out of 24 isolates produced sclerotia in soil and the number and size of sclerotia varied considerably among isolates within the same AG. Sclerotia from each group ranged in size from 0.25 mm to more than 2.5mm dia, with most between 0.5–1.0mm dia. Sclerotia formed in soil were smaller than those formed in pure culture. Sclerotia of each AG were tested for viability after 30–270 days incubation in artificially-inoculated soil. Sclerotia produced in soil were brown to dark-brown and these were divided into two distinct groups on external morphology: firm and irregularly globose to subglobose with either a pitted or smooth surface in one group, and rather soft with an indefinite shape in the other. On the basis of internal morphology, the sclerotia fell into three groups. Sclerotia of AG-1 were composed of three well-defined layers; the central cells had dense contents and were surrounded by an outer layer of empty cells which were bordered by a darkly-pigmented mucilaginous surface-layer. Sclerotia of AG-2, Type-1 had only two well-defined layers; The darkly-pigmented mucilaginous layer was absent. Sclerotia of the other AG were constructed of very loosely-arranged brown cells without any well-defined zones.     

Bovine haptoglobin: single radial immunodiffusion assay of its polymeric forms and dramatic rise in acute-phase sera.

Using purified bovine haptoglobin (Hp) and specific antisera, a single radial immunodiffusion (SRID) assay method has been developed to measure the serum Hp level in cattle. Bovine Hp is a highly polymerized protein showing heterogeneous molecular forms in serum. After treatment with cysteine or glutathione, Hp was partially reduced to a homogeneous form, suitable for SRID assay. This method gives values comparable to those obtained by hemoglobin-binding capacity assay, and has the advantage of being simple and convenient. Although serum Hp was not detectable in healthy cattle, it was found more than 50-fold after invasive surgery, indicating that Hp is a characteristic acute-phase protein in cattle.     

Physiological and Mineralogical Properties of Arsenic-Induced Chlorosis in Barley Seedlings Grown Hydroponically

The experiment was carried out to investigate the effects of arsenic (As) on the physiological and mineralogical properties of barley (Hordeum vulgare L. cv. ‘Minorimugi’). The plants were grown in nutrient solution treated with 0, 6.7, 33.5, and 67 μ M As (0, 0.5, 2.5, and 5 ppm As, respectively) in the phytotron. Dry matter yield of shoots and roots decreased significantly with the As treatments, indicating that barley plants are As-sensitive and As-toxicity depends on the As concentration in the rooting medium. Necrosis in older leaves and chlorosis symptoms (whitish color) in the fully developed young leaves were observed at the 33.5 and 67 μ M As treatments. Arsenic concentration, accumulation, and translocation increased with the increase of As concentration in the rooting medium. Arsenic was mostly concentrated in roots and a little amount was moved to shoots, indicating that As was not easily translocated to shoots of barley seedlings. Concentrations and accumulations of phosphorus (P), potassium (K), calcium (Ca), magnesium (Mg), manganese (Mn), zinc (Zn), and copper (Cu) decreased significantly in shoots for 33.5 and 67 μ M As treatments as compared to the 0 μ M As treatment. Concentrations of P, K, Ca, Mg, Mn, and Cu decreased in roots, but Zn concentration increased in roots at 67 μ M As treatment. Accumulations of P, K, Ca, Mg, Mn, Zn, and Cu in roots also decreased significantly at 67 μ M As treatment. Accumulation of P and the cations showed negative relationship with As. Concentration of Fe decreased in shoots at 33.5 and 67 μ M As treatments where chlorosis was induced in the young leaf but increased in roots at 33.5 and 67 μ M As treatments. It was suggested that As might induce iron (Fe)-chlorosis in the plants. Among the micronutrients, Fe translocation was more affected than others by As. Phytosiderophore (PS) accumulation in roots, which is a symptom of Fe-deficiency in grasses, did not change significantly between 0 and 33.5 μ M As treatments; indicating that As-induced chlorosis did not enhance PS accumulation in roots and decreased due to As-toxicity at 67 μ M As treatment.     

Isolation of canine C-reactive protein and characterization of its properties

C-reactive protein (CRP) was isolated from the acute phase serum of dogs subjected to surgical stimulation. Its properties were characterized. Canine CRP was isolated by ion-exchange chromatography using DEAE-Sephacel and DEAE-Sephadex A-50 and affinity chromatography using protein A-Sepharose CL 4B in combination with agar-block electrophoresis. In immunoelectrophoresis, canine CRP had the same γ-mobility as human γ-type CRP. The molecular weight of purifined canine CRP was estimated by gel filtration and sodium dodecyl sulfate polyacrylamide gel electrophoresis to be approximately 157 000 and 155 000 respectively. This CRP was a thermolabile protein which completely lost its antigenicity by heating at 70°C for 15 min. The serum concentration of CRP in normal beagle dogs ranged from 0.198 to 0.826 μg ml⁻¹ (0.486±0.170 μg ml⁻¹). The concentration was acutely increased by surgery as it was in man and was rapidly decreased with convalescence. Dogs can be a useful animal model for investigation of the mechanism of CRP production and the function of CRP.     

Heterogeneity of Hanganutziu-Deicher antigen glycoproteins in different species animal sera.

A heterophilic Hanganutziu-Deicher (HD) antigen is present in many animal sera except human and chicken sera. To visualize the antigenic molecules, nine species animal and human sera were analyzed by SDS-PAGE, followed by Western blotting and immunostaining with avain anti-N-glycolylneuraminyl-lactosyl ceramide antibody which recognizes the terminal N-glycolylneuraminic acid moiety of glycoconjugates as an epitope of the HD antigen. Several HD antigen-active glycoprotein bands were detected in the sera of fetal calf, calf, horse, goat, monkey, rabbit, guinea pig, rat and mouse, except for human serum. The HD antigenic proteins showed heterogeneities in their molecular weights and were not identical with any major band visualized with silver-staining, indicating that they are minor components of serum proteins in each animal. Neuraminidase treatment destroyed the antigenicity of all proteins, confirming that N-glycolylneuraminic acid (NeuGc) at the non-reducing terminal of carbohydrate chains is the antigenic epitope in serum glycoprotein molecules as already confirmed in glycosphingolipid (GSL) antigens. The finding of HD-antigenic glycoproteins in animal sera suggests that they also stimulate HD antibody production in patients who received animal antiserum for therapeutic aim.     

Isolation and characterization of C-reactive protein and serum amyloid P component from bovine serum

C-reactive protein (CRP) and serum amyloid P component (SAP), which are known as acute phase reactants in human and many other animals, were purified from cow sera. Affinity chromatography using HE agarose gel was the most effective method to isolate both CRP and SAP from a large volume of bovine serum. Separation of CRP and SAP from the mixed preparation could be performed by DEAE-cellulose column chromatography, gel permeation HPLC using TSK-G3000SW or affinity chromatography using phosphorylcholine derivatives of bovine serum albumin-conjugated Toyopearl HW 65. Bovine CRP and SAP were identified as genuine CRP- and SAP-class proteins by their cross reactivities with anti-human CRP and anti-human SAP, respectively, and by their homology in amino acid compositions compared with those of human CRP and SAP, respectively. Bovine CRP moved slower than beta-globulin, and bovine SAP moved in the beta-globulin region in agarose gel electrophoresis. Both of them gave single bands in native polyacrylamide gel electrophoresis (PAGE). Bovine CRP and SAP molecular weights were estimated to be 100,600 and 109,500 daltons respectively, by sedimentation equilibrium analysis. Bovine CRP showed 23K dalton subunits by sodium laurylsulfate-PAGE and bovine SAP showed 28K and 32K dalton subunits, both of which were glycosylated and had identical amino acid compositions, indicating that both CRP and SAP molecules are pentamers. In fact, they appeared to have pentameric disk-like configurations in electronmicroscopical examination.