Propolypeptide of von Willebrand factor circulates in blood and is identical to von Willebrand antigen II

The generally mild bleeding disorder of von Willebrand disease is associated with abnormalities of two distinct plasma proteins, the large multimeric von Willebrand factor (vWF), which mediates platelet adhesion, and von Willebrand antigen II (vW AgII), which is of unknown function. The two proteins were found to have a common biosynthetic origin in endothelial cells and megakaryocytes, which explains their simultaneous absence in the severe form of this hereditary disease. Shared amino acid sequences from a 100-kilodalton plasma glycoprotein and from vW AgII are identical to amino acid sequences predicted from a complementary DNA clone encoding the 5' end of vWF. In addition, these proteins have identical molecular weights and immunologic cross reactivities. Monoclonal antibodies prepared against both proteins recognize epitopes on the pro-vWF subunit and on a 100-kilodalton protein that are not present on the mature vWF subunit in endothelial cell lysates. In contrast, polyclonal antibodies against vWF recognize both pro-vWF and vWF subunits. Thus, the 100-kilodalton plasma glycoprotein and vW AgII are identical proteins and represent an extremely large propolypeptide that is first cleaved from pro-vWF during intracellular processing and then released into plasma.     

Atomic-Scale Images of the Growth Surface of Ca1-xSrxCuO2 Thin Films

The surface microstructure of c-axis (Ca,Sr)CuO(2) thin films, grown by laser molecular beam epitaxy on SrTiO(3)(001) substrates, was studied by ultrahigh-vacuum scanning tunneling microscopy (STM). Images were obtained for codeposited Ca1-xSrxCuO(2) thin films, which show a layered-type growth mode. The surfaces consist of atomically flat terraces separated by steps that are one unit cell high. A pronounced dependence of the growth mechanism on the Sr/Ca ratio of the films was observed. Atomic resolution STM images of the CuO(2) sheets in the ab plane show a square lattice with an in-plane spacing of 4 angstroms; the lattice contains different concentrations of point defects, depending on the polarity of the sample-tip bias.     

Mitogenesis in response to PDGF and bombesin abolished by microinjection of antibody to PIP2

The turnover of phosphatidylinositol 4,5-bisphosphate (PIP2) is believed to constitute a crucial step in the signaling pathways for stimulation of cells by a variety of bioactive substances, including mitogens, but decisive evidence for the idea has not been obtained. In the present study, a monoclonal antibody to PIP2 was microinjected into the cytoplasm of NIH 3T3 cells before or after exposure to mitogens. The antibody completely abolished nuclear labeling with [3H]thymidine induced by platelet-derived growth factor and bombesin, but not by fibroblast growth factor, epidermal growth factor, insulin, or serum. The findings strongly suggest that PIP2 breakdown is crucial in the elicitation and sustaining of cell proliferation induced by some types of mitogens such as platelet-derived growth factor and bombesin.     

Comparative pathogenicity study on antigenically variant strains of Enterococcus seriolicida

Enterococcus seriolicida strains were divided into two groups, agglutinating and nonagglutinating, by a slide agglutination test using antiserum against the YT-3 strain. Intraperitoneal injection of agglutinating and nonagglutinating strains into yellowtail, Seriola quinqueradiata Temminck & Schlegel, revealed that nonagglutinating strains were more virulent than agglutinating strains. Two nonagglutinating and highly pathogenic strains SS91–014 and SS91–092 were subcultured 30 times in brain heart infusion broth, and the agglutination titres of 50 colonies from subcultures 1, 2, 3, 4, 5, 6, 11, 16, 21, 26 and 30 against anti-YT-3 serum were determined. Transformation from a nonagglutinating (1:<4) to an agglutinating (1: ≥4) pattern was first observed at the sixteenth subculture, and the ratio of agglutinating to nonagglutinating substrains rose until the thirtieth subculture. At this time, 70% of the SS91–014 population and 52% of the SS91–092 population were transformed to an agglutinating pattern. When the pathogenicity of four transformed substrains with different agglutination titres was tested in yellowtail, the nonagglutinating substrain showed higher pathogenicity than the agglutinating substrains, but no relationship between LD₅₀ values and the agglutination titres of transformed substrains was observed. The pathogenicity of E. seriolicida appears to be related to the agglutination pattern, although it was not demonstrated that this property is solely responsible for pathogenicity.     

Estimating the spawning season of black sea bream Acanthopagrus schlegelii in Hiroshima Bay,Japan, from temporal variation in egg density

Fisheries Science - We aimed to determine the spawning period of black sea bream Acanthopagrus schlegelii in Hiroshima Bay, Japan, based on the distribution of its eggs, to assist in the resource...     

Establishment and characterization of two new cell lines derived from flounder, Paralichthys olivaceus (Temminck & Schlegel)

Two new cell cultures from flounder, Paralichthys olivaceus (Temminck & Schlegel), flounder fin (FFN) cells from fin tissue and flounder spleen (FSP) cells from spleen tissue, were established and characterized. The cells multiplied well in Eagle's minimum essential medium, supplemented with 10% foetal bovine serum, and have been subcultured more than 100 times, becoming continuous cell lines. Modal diploid chromosome number of FFN and FSP cells was 64 and 62, respectively. Polymerase chain reaction products were obtained from FFN and FSP cells with primer sets ofmicrosatellite markers of flounder. Optimal growth temperature was 20 degrees C and consisted of epithelioid cells. FFN and FSP cells showed cytopathic effects after inoculation of infectious pancreatic necrosis virus, marine birnavirus, chum salmon virus, infectious haematopoietic necrosis virus, spring viraemia of carp virus and hirame rhabdovirus. Thus these new cell lines may be useful for studying a wide range of fish viruses.     

Characterization of wood-based molding bonded with citric acid

The wood-based moldings were fabricated by using only citric acid as an adhesive. The mechanical properties, water resistances, thermal properties and chemical structure were investigated. Wood powder obtained from Acacia mangium was mixed with citric acid under certain weight ratios (0–40 wt%), and each powder mixture was molded using two types of metal molds at 200°C and 4 MPa for 10 min. The modulus of rupture and the modulus of elasticity values of the wood-based molding containing 20 wt% citric acid were 35.8 MPa and 5.4 GPa, respectively. The maximum impact strength was 0.94 kJ/m² with the same citric acid content. The water resistance increased with increasing citric acid content, and the good resistance to boiling water was also recognized. However, some elution of substances derived from citric acid was observed at high levels of citric acid, causing a decrease in the thermal properties. The ester linkages were detected by Fourier transform infrared spectroscopy, indicating that the citric acid reacted with the wood. Our results demonstrated that citric acid brought about good adhesiveness for fabricating the wood-based molding.     

Susceptibilities against bovine lactoferrin with microorganisms isolated from mastitic milk

Antibacterial effects of bovine lactoferrin were studied in vitro against microorganisms isolated from mastitic milk in Tokachi area, Hokkaido, Japan. Microorganisms isolated were Escherichia coli (11 isolates), Klebsiella pneumoniae (5 isolates), enterococci (8 isolates), Staphylococcus aureus (10 isolates), coagulase negative staphylococci (CNS, 13 isolates), streptococci (11 isolates), Prototheca zopfii (7 isolates) and yeast-like fungi (9 isolates). Lactoferrin has been known as a multifunctional protein and its antimicrobial effect is one of the most essential function of it. In order to compare their susceptibilities against lactoferrin, the minimal inhibitory concentration values were estimated by a microplate assay method using 96-well microplate, which involved measuring the optical density of the cultures. Prototheca zopfii was highly sensitive to bovine lactoferrin and complete inhibition of this microorganism was observed even at the low concentration of 7 mug/ml. On the other hand, E. coli and enterococci showed resistance against lactoferrin action and staphylococci showed strain-dependent resistance.     

Dynamics of lingual antimicrobial peptide,lactoferrin concentrations and lactoperoxidase activity in the milk of cows treated for clinical mastitis

The aim of the present study was to examine changes in innate immune factors in the milk of mastitic dairy cows treated with antibiotics. Cows in the antibiotics group (n = 13) were infused into the mammary gland with cefazolin on the sixth day after mastitis was diagnosed (the day of the mastitis diagnosis = day ?6). The control group (n = 12) was not treated. Milk samples were collected once every 2 days from days ?6 to 12 and somatic cell count (SCC), lingual antimicrobial peptide (LAP), and lactoferrin (LF) concentrations and lactoperoxidase (LPO) activity were measured. SCC and LF concentrations in the antibiotics group markedly decreased after the antibiotic treatment. When cows in the antibiotics group were divided according to SCC on day 0, LAP concentrations and LPO activity in cows with a lower SCC on day 0 (<5 × 10⁶ cell/mL) were significantly higher and lower than those in cows with a higher SCC, respectively. These results suggest that LF concentration decreased with decrease in SCC after treatment and that LAP concentration and LPO activity differed depending on the severity of mastitis. This is the first report to reveal the dynamics of innate immune factor in milk of cows treated for clinical mastitis.     

Central nervous system lesions caused by canine distemper virus in 4 vaccinated dogs

We examined the cerebellum and cerebrum of 4 vaccinated dogs, 3–60-mo-old, that displayed clinical signs of canine distemper virus (CDV) infection, and died 7–40 d after developing neurologic signs. The main histologic lesions were demyelination, gliosis, meningitis, perivascular lymphocytic cuffing, and inclusion bodies. These lesions were similar in all 4 cases regardless of the time since vaccination, except that meningoencephalitis and gliosis were subacute in 3 dogs and chronic in 1 dog. However, these differences did not appear to be related to their vaccination status. Immunohistologically, a CDV-positive immunoreaction was seen mainly in astrocytes, neurons and their axons, lymphocytes around and in the blood vessels of the pia mater and choroid plexus, ependymal cells of each ventricle, and the cells of the choroid plexus. The histologic and immunohistologic changes were similar in the cerebellum and cerebrum. The genetic characterization of the virus strains in 2 of these naturally occurring canine distemper cases confirmed that they were South American wild-type strains (Kiki and Uy251) belonging to the EU1/SA1 lineage. These strains are not included in the commercial CDV vaccines available in Uruguay.