Isolation and characterization of aminopeptidase (Jc-peptidase) from Japanese cedar pollen (Cryptomeria japonica)

An aminopeptidase, Jc-peptidase, was purified from Japanese cedar pollen by seven steps, including precipitation with ammonium sulfate, ion-exchange chromatography, gel filtration, hydrophobic interaction chromatography on phenyl-agarose, and high-performance liquid chromatography. Purified Jc-peptidease has a molecular weight of 42 kDa and hydrolyzes the synthetic substrates of L-phenylalanyl-4-methylcoumaryl-7-amide (Phe-MCA) with Km = 5 x 10(-5) M, Tyr-MCA with Km = 7 x 10(-4) M, Leu-MCA with Km = 1 x 10(-3) M, and Met-MCA with Km = 1 x 10(-3) M. Other MCA analogues such as Arg-MCA or Glu-MCA failed to serve as its substrates. The activity was inhibited in the presence of phebestin, [(2S,3R)-3-amino-2-hydroxy-4-phenylbutanoyl-L-valyl]-L-phenylalanine, with Ki = 4.7 x 10(-5) M, or bestatin, [(2S,3R)-3-amino-2-hydroxy-4-phenylbutanoyl]-L-leucine, with Ki = 1.1 x 10(-4) M. According to amino acid sequence analysis, the N-terminal amino group seems to be blocked. The physiological function of the aminopeptidase (Jc-peptidase) has not been clarified in vivo.     

Fat depot‐specific differences in leptin mRNA expression and its relation to adipocyte size in steers

ABSTRACT This experiment was conducted to investigate leptin mRNA expression, adipocyte size, and their relationship in several adipose tissues of fattening steers. Subcutaneous, perirenal, intermuscular and intramuscular adipose tissues were collected from three crossbred steers (Japanese Black cattle X Holstein) aged 21 months. The mRNA level and adipocyte diameter were determined in these adipose tissues. The intramuscular adipose tissue had a lower leptin mRNA level than the intermuscular and perirenal adipose tissues (P < 0.05). Leptin mRNA level was lower in the subcutaneous depot than in the intermuscular depot (P < 0.05). Adipocyte diameter was larger in the intermuscular adipose tissue than in the subcutaneous and intramuscular adipose tissues (P < 0.05). Leptin mRNA level was positively correlated with adipocyte diameter (r² = 0.81, P < 0.05). These results suggest that the cattle have fat depot‐specific differences in leptin gene expression, which are a result of a difference in adipocyte size.     

‘巨峰’葡萄主梢和副梢花器官与种子发育的研究

 ‘巨峰’葡萄主梢花粉粒大于副梢, 花粉染色率与副梢差异不显著, 发芽率低于副梢, 但花粉发芽率不影响种子的形成。主梢小花雌蕊在减数分裂后形成的功能大孢子小于副梢。同时开花前主梢胚囊的发育状况优于副梢, 开花期正常胚囊率主梢也明显低于副梢。因此, 开花前后主梢异常胚囊的出现是导致受精发生障碍并使结籽率低下的原因。     

Effect of the strain combination of the donor and recipient on the production efficiency of W‐bearing sperm in mixed‐sex germline chimeric chickens

Effect of the strain combination of the donor and recipient on production efficiency of W‐bearing sperm in mixed‐sex chimeric testes was analyzed. The combinations of the donors and recipients were White Leghorn (WL) and Rhode Island Red (RIR), and vice versa. Generated mixed‐sex chimeras that had the male phenotype at sexual maturity were classified into four groups: (1) a female WL donor and a male RIR recipient; (2) a male WL donor and a female RIR recipient; (3) a female RIR donor and a male WL recipient; (4) a male RIR donor and a female WL recipient. The mean number of W‐bearing sperm detected by in situ hybridization among 10 000 sperm observed were 147, 165, 30 and 45 in groups 1, 2, 3 and 4, respectively. The numbers in groups 1 and 2 were both significantly higher than those of groups 3 and 4 (P < 0.05). The combination of a WL donor and a RIR recipient produced W‐bearing sperm more efficiently than the reverse combination.     

Preparation and characterization of monoclonal antibodies against an avian reovirus

Thirteen monoclonal antibodies against avian reovirus strain Uchida were derived. Of the 13 antibodies, three (MAb1, MAb2, and MAb3) had the ability to neutralize the infectivity of the virus. MAb1 neutralized strains Uchida, CS-108, and TS-142 equally. MAb2 neutralized the same three strains, but the activity of neutralization was 10 times higher against Uchida than against CS-108 and TS-142. MAb3 neutralized only strain Uchida. It seems that MAb1 and MAb2 have a rather broad neutralization activity and that MAb3 has a type-specific activity. These data indicate that there are both type-specific and more broadly specific neutralization epitopes in avian reovirus particles, as in mammalian reoviruses. In Western blot analysis, MAb1 bound to a sigma protein of strain Uchida, so it was suggested that this protein carries the epitope of the less specific neutralizing activity.     

Changes in Activities of Enzymes Related to Energy Metabolism in Peripheral Leukocytes of Fattening Steers

Glucose, triglyceride, cholesterol and immunoreactive insulin (IRI) concentrations, some enzyme activities in plasma, and activities of enzymes related to energy metabolism in peripheral leukocytes were measured in fattening Japanese Black Wagyu x Holstein steers fed on different diets at 8, 12, 16, 20 and 24 months of age. The plasma IRI concentrations at 20 and 24 months of age were significantly higher than those at 8 months of age. Activities of hexokinase (HK), glucose-6-phosphate dehydrogenase (G6PD), aspartate aminotransferase (AST), and malate dehydrogenase (MDH) in cytosolic fractions, and glutamate dehydrogenase (GLDH), MDH and AST in mitochondrial fractions in peripheral leukocytes of steers at 24 months of age were significantly higher than those at 8 months. Increasing plasma insulin concentration was considered to induce acceleration of glucose utilization in leukocytes of fattening steers. The cytosolic ratio of MDH/lactate dehydrogenase (LDH) activity in leukocytes increased significantly in the fattening process and was considered to be a useful indicator for evaluating changes in energy metabolism in steers.     

Ability of CD8+ T cell anti-feline immunodeficiency virus (FIV) activity and FIV proviral DNA load in mononuclear cells in FIV-infected cats

We investigated the relationship between CD8+ T cell anti-feline immunodeficiency virus (FIV) activity and FIV proviral DNA load integrated in mononuclear cells. The anti-FIV activity and the proviral DNA load were correlated, and the number of proviral DNA copies was high in cats with decreased anti-FIV activity. Particularly, no anti-FIV activity was detected in the cats staged as having an acquired immunodeficiency syndrome (AIDS)-related complex or AIDS, and the number of proviral DNA copies was obviously increased compared to those in the cats in the asymptomatic stage. These results suggest that decreased anti-FIV activity destroys the control of in vivo FIV replication, which leads to an increased proviral DNA load with the progression of the clinical stage of disease.     

Production and characterization of monoclonal antibodies against porcine interleukin-4

Interleukin 4 (IL-4) is an important regulatory cytokine produced by activated T lymphocytes and mast cells, and regulates the growth and differentiation of cells such as B and T lymphocytes. In the present study, recombinant thioredoxin (Trx)-porcine IL-4 (pIL-4) fusion protein was prepared by Escherichia coli (E. coli), and by using this protein as an immunogen, monoclonal antibodies (mAbs) against pIL-4 were produced to establish a basis for a research on immune responses in pigs. Six stable hybridoma cell lines were successfully established and specific binding of each mAb to recombinant pIL-4 produced by E. coli and insect cells infected with recombinant baculovirus was shown by enzyme-linked immunosorbent assay (ELISA) and/or immunoblot analysis. Isotype analyses of these mAbs revealed that the subclass of 5 out of 6 mAbs was IgG1 and the rest was IgG2b. Further, assessment of their epitopes by competition binding assay indicated that the mAbs obtained in this study bound to 4 different epitopes. The recombinant proteins and mAbs produced in this study will be useful tools for the assessment of porcine immune system.