Shape changes in caprine erythrocytes exposed to chlorpromazine and lysolecithin

Stomatocytic and echinocytic transformations of caprine erythrocytes were studied in vitro using chlorpromazine as a stomatocyyic agent and lysolecithin as an echinocytic agent. Morphologic changes in erythrocytes generally varied with the cell shape and the concentration of the substances used. Discoytic, triangular, and pear-shaped red cells common to normal goats, exhibited classical stomatocytic and echinocytic changes with formation of spherostomatocytes, sphero-echinocytes, and spherocytes. In comparison, fusiform and spindle-shaped red cells found in certain Angora goats seemed less prone to shape changes and required greater concentrations of the inducing agents to effect such changes. Chlorpromazine at higher concentrations also inflicted localized membrane damage in form of tiny pits and lysolecithin likewise induced formation of fragile smooth or beaded filaments.     

Effect of a single polymorphism in the Japanese quail NK‐lysin gene on antimicrobial activity

NK‐lysins are cationic peptides that play important roles in host protection, and are an important constituent of innate immunity. We identified nine single‐nucleotide polymorphisms (SNPs) in the NK‐lysin open reading frame (ORF) from 32 Japanese quails in six strains: A, B, ND, K, P, and Y. The G to A substitution at nucleotide position 272 in the ORF resulted in a Gly (G) to Asp (D) amino acid substitution (Cj31G and Cj31D alleles). The Cj31D allele was detected in P (frequency 0.76) and Y (frequency 0.03) strains. We compared the antimicrobial activities of four synthetic peptides from the helix 2‐loop‐helix 3 region of avian NK‐lysins against Escherichia coli: Cj31G and Cj31D from quail and Gg29N and Gg29D from chicken. The antimicrobial activities of the four peptides decreased in the following order: Gg29N > Cj31G > Gg29D > Cj31D (P < 0.05). Although there were no differences in the predicted secondary structure of the Cj31G and Cj31D, the net charge of the Cj31G was higher than that of Cj31D. These data indicated that the antimicrobial activity of CjNKL is influenced by net charge, similar to that which has been observed in chicken. © 2015 Japanese Society of Animal Science     

In vitro platelet release by rat megakaryocytes: effect of metabolic inhibitors and cytoskeletal disrupting agents

Development of an in vitro visual assay facilitated the study of large numbers of megakaryocytes undergoing proplatelet formation in short-term cultures. Approximately 9% of megakaryocytes formed platelets during a 24-hour period. In the presence of an inhibitor of anaerobic glycolysis (NaF), proplatelet formation was inhibited, whereas inhibitors of respiration (NaCN) did not significantly (P greater than 0.05) decrease proplatelet formation. Presence of the microtubule-disrupting agents colchicine and vincristine sulfate in culture medium inhibited proplatelet formation, whereas the microfilament-disrupting agent cytochalasin B had a less pronounced inhibition.     

Methods for detection of immune-mediated neutropenia in horses, using antineutrophil serum of rabbit origin

Equine neutrophil antibody was raised in rabbits inoculated with equine neutrophils isolated to purity greater than 99.0%, using Percoll density-gradient sedimentation. Neutrophil antibody was detected by use of agar gel diffusion, leukoagglutination, indirect immunofluorescence, staphylococcal protein A and streptococcal protein G binding, and phagocytic inhibition techniques. Precipitin lines and leukoagglutination were seen in antiserum dilutions of 1:4 and 1:64, respectively. The specific nature of leukoagglutination was characterized by the formation of rosette-like clumps of neutrophils. Specific bright membranous fluorescence was seen in neutrophils treated with the antiserum and exposed to fluorescein-conjugated goat anti-rabbit immunoglobulin, and staphylococcal protein A and streptococcal protein G. Whereas the indirect immunofluorescence and protein G-binding tests were equally sensitive and resulted in titer of 1:256, the protein A-binding test was less sensitive and resulted in titer of only 1:32. Nonspecific binding of protein A and protein G was noticed as uniform or patchy cellular fluorescence in a small number of neutrophils. Treatment of neutrophils with antiserum up to dilution of 1:8 resulted in a significant (P less than 0.05) suppression of phagocytosis of opsonized zymosan particles. Thus, protein G-binding and indirect immunofluorescence tests are highly sensitive to detect neutrophil antibody and may be used to diagnose immune-mediated neutropenias in horses and, possibly, in other animal species.     

Developmental patterns of lamellae and stroma fractions of chloroplasts in rice plants

On examining the changes in lamellae and stroma nitrogen during leaf development, it is demonstrated that the lamellae and stroma fractions ofrice chloroplasts develop in quite different ways. In the case of stroma, the stroma materials existing in the leaf section which has just emerged from a leaf sheath are quite limited and the major part of this fraction is derived from the successive protein synthesis, i.e., the synthesis of this fraction was markedly increased during leaf expansion. This developmental pattern of the stroma coincided with the changes in the high-molecular-weight water soluble leaf protein, which seemed to be mainly composed of Fraction I protein. A rapid increase in stroma nitrogen was found to be a major cause for an increase in the leaf nitrogen content during leaf development.

On the other hand, the developmental pattern of the lamellae fraction was characterized by the fact that a considerable amount of this fraction had already been prepared when a leaf emerged from a leaf sheath and thereafter, no outstanding increase was seen compared to that of the stroma. This developmental pattern of the lamellae fraction resulted in a lowering of the proportion of lamellae nitrogen to the total leaf nitrogen during leaf development.

A great change in the lamellae-stroma composition of chloroplasts was observed. The proportion of stroma nitrogen to the total chloroplast nitrogen tended to increase as a leaf develops. Since the developmental stage varied according to the regions of a leaf, variation of the lamellaestroma composition was seen even within a leaf, i.e., the proportion of stroma nitrogen increased from base to tip.

In order to compare the synthetic rate of chlorophyll with those of the stroma and lamellae fractions, the changes in the ratios of stroma nitrogen/chlorophyll and lamellae nitrogen/chlorophyll were examined. The lamellae nitrogen/chlorophyll ratio decreased as a leaf developed, whereas the stroma nitrogen/chlorophyll ratio increased. Then the synthetic rates of these fractions during leaf development turned out to be of the same order as the stroma fraction, chlorophyll, lamellae fraction.     

Identification and characterization of hens transmitting avian leukosis virus (ALV) to their embryos by ELISAs for detecting infectious ALV, ALV antigens and antibodies to ALV.

A total of 72 White Leghorn grandparent hens was examined by ELISA for avian leukosis virus (ALV), ALV antigens and anti-ALV antibodies to identify and characterize the hens transmitting ALV to their embryos (transmitters) by using fertilized eggs. These hens were divided into 3 groups as no antibody and non-viremic (NANV) (49 hens), antibody-positive and non-viremic (APNV) (21 hens) and no antibody and viremic (NAV) (2 hens) by testing the sera for the presence of ALV and anti-ALV antibody. Egg albumen and embryos were tested for the presence of ALV and ALV antigens. As a result, no ALV was detected in both albumen and embryos in the NANV group. On the other hand, all albumen samples collected repeatedly from 3 hens of the APNV group and 2 hens of the NAV group contained infectious ALV, although the infectivity differed with the individual. Also, these 5 hens produced infected embryos at varying frequencies. However, on AP hen which shed neither ALV nor ALV antigens into the albumen produced an infected embryo at a lower rate. These results indicate that testing for infectious ALV in albumen from a newly laid egg per hen is effective to identify the transmitters to some extent. When virus titers in each of 8 tissue samples from the 6 transmitting hens were determined, the highest virus titers were found in washing from the ampulla of the oviducts in most of the shedders, suggesting that embryo infection is closely correlated with ALV produced at the oviduct, but not with ALV transferred from the other parts of the body.     

Control of clubroot of crucifers by Phoma glomerata and its product epoxydon

Phoma glomerata strain JCM9972 controls clubroot of cruciferous crops caused by Plasmodiophora brassicae and the activity depends upon epoxydon (5-hydroxy-3-(hydroxymethyl)-7-oxabicyclo [4.1.0]hept-3-en-2-one) produced by the strain.