Anti-rat myoglobin antisera in the immunocytochemical diagnosis of rhabdomyosarcomas of rats

Anti-rat myoglobin (Mb) was prepared and used in the avidin-biotin-peroxidase complex (ABC) method on paraffin-embedded sections of nine soft tissue tumors (including two rhabdomyosarcomas) of rats. Distribution and nature of the reactive substance to Mb antiserum were compared to those of desmin antiserum. Rat Mb was isolated from the skeletal muscle; monospecificity of the rat antiserum was confirmed by the immunoblotting procedures. The Mb antiserum reacted specifically to normal and neoplastic striated muscle cells. Mb-staining reactions were present diffusely in the cytoplasm, while desmin-staining substances were localized at Z-bands or were diffuse in the cytoplasm as separated aggregates. Reaction to the Mb serum was also detected in cells of thick portions of Henle's loop and distal convoluted tubules.     

Physico-chemical properties of calf-diarrhea coronavirus

Replication of calf diarrhea coronavirus was not inhibited by 5-iodo-2′-deoxyuridine, indicating that the virus is an RNA virus. Sensitivity to ether and chloroform indicated that the virus is enveloped, and this was confirmed by electron microscopic observation of the virion. The virus was readily inactivated by trypsin and sodium deoxycholate. The virus was labile at 50°C in diluted medium, but readily stabilized in the presence of MgCl₂. It was stable at pH 5 and 7, while a slight loss of infectivity was observed at pH 3. The virus was readily filtered through membrane filters of 200 and 100-nm pore sizes, but not through 50-nm filters. The buoyant density of the virion in CsCl was estimated to be 1.25 g/ml.     

Establishment of a large-scale purification procedure for purified recombinant bovine interferon-tau produced by a silkworm-baculovirus gene expression system

We developed a procedure for the large-scale purification of bovine interferon-tau (boIFN-tau) by means of a silkworm-baculovirus gene expression system. Recombinant boIFN-tau (rboIFN-tau) was efficiently produced in the silkworm infected with boIFN-tau cDNA recombinant baculovirus and accumulated in the haemolymph. To establish a purification method suitable for mass production, we tried three crude purification methods, namely, an acidification and neutralization treatment (ANT), silica gel column chromatography (SGCC), and Blue sepharose column chromatography (BSCC) with a combination of Q-sepharose (QSC) and chelating sepharose column chromatographies (CSCC). As a result, the acidification and neutralization treatment was found to be the most efficient and cost effective. With this combination, we obtained 91% pure products. To confirm the applicability of the procedure for mass production, we inoculated 100 silkworms with the recombinant virus, and recovered about 4.55 mg (1.26 x 10(8)U/mg) of 91% pure rboIFN-tau by means of a combination of the ANT, followed by QSC and CSCC.