Inhibitory effects of pesticides on growth and respiration of cultured cells

Effects of various drugs including pesticides on the growth and respiration of cultured cells were evaluated comparatively using cell lines derived from mosquito ovary and subcutaneous mouse tissues. The concentration producing 50% inhibition of cell growth, I₅₀ (M), was determined for each of 42 drugs. Inhibitors of respiration and nucleic acid and protein biosyntheses such as rotenone, piericidin A₁, actinomycin D, and puromycin had very high pI₅₀ values of approximately 8. Except for the compounds known to be uncouplers of oxidative phosphorylation, the drugs suppressed the respiration rate of the cultured cells to various degrees. The pI₅₀ value (and the pEC₁₅₀, 150% enhancement of the control, value for uncouplers) was determined for each compound. By examining the relation of pI₅₀ (and pEC₁₅₀) values between cell growth and respiration, the compounds could be classified into two groups according to their modes of inhibitory action against the cultured cells. One mode relates to the inhibition of energy synthesis and the other, perhaps, to interference with the biosynthesis of biomacromolecules.     

N-acetylglucosaminyltransferase I activity of bovine oviduct epithelial cells: stimulation by luteinizing hormone, vascular endothelial growth factor and tumor necrosis factor alpha

N-acetylglucosaminyltransferase I (GnT I; EC 2.4.1.101), which catalyzes the first step in the conversion of oligomannose to complex or hybrid N-glycans of glycoproteins, was found in media cultured with bovine oviduct epithelial cells (BOEC) obtained from non-pregnant cows during the follicular phase. Combined treatment with specific hormones increased GnT I release from BOEC. Luteinizing hormone (LH; 10 ng/ml) alone slightly, but together with 17beta-estradiol (E2; 1 ng/ml), synergistically increased GnT I activity. Vascular endothelial growth factor (VEGF) and tumor necrosis factor (TNF) alpha, which have been shown to have their highest activities in the bovine oviduct during the periovulatory period, also increased in GnT I activity. This study provides the first evidence of an increase of GnT I release from BOEC in vitro, and shows that endocrine as well as local factors such as LH, VEGF and TNFalpha increase this activity. The results suggest that GnT I activity in the bovine oviduct may contribute to the induction of glycosylation and thereby contributing to the provision of the optimal microenvironment for fertilization and early development of the embryos.     

Recovery of the everted cystacanth of seven acanthocephalan species of birds from feral raccoons (Procyon lotor) in Japan

Acanthocephalan everted cystacanths were detected in the intestines of 61 out of 555 feral raccoons (Procyon lotor) trapped between May 2003 and April 2005 in the western region of mainland Japan (Honshu). All collected specimens were identified as the species of birds including 3 Centrorhynchus species (C. bazaleticus, C. elongatus, and C. teres), Sphaerirostris lanceoides, Plagiorhynchus ogatai, Porrorchis oti, and Southwelina hispida. Three species, C. bazaleticus, C. teres, and S. lanceoides, were new to distribution records in Japan and the Far East. Recovery rates of these acanthocephalans were higher from the raccoons trapped in spring and early summer (range 8.3-36.8%, average 21.5%) than during the remainder of the year (range 0-20.0%, average 6.3%). The reason for this seasonal change is unknown. There is little information on the acanthocephalan fauna of wild birds in Japan, however, recovery of the cystacanths from the feral raccoons captured through the year might provide useful information on the fauna and geographical distribution of avian acanthocephalans.     

Antigenic analysis of avian Chlamydia psittaci using monoclonal antibodies to the major outer membrane protein.

Monoclonal antibodies to the major outer membrane protein (MOMP) of Chlamydia psittaci derived from a parrot were established for antigenic analysis of avian C. psittaci. With 17 monoclonal antibodies to MOMP, 17 reactivity patterns were identified on 112 strains of C. psittaci, C. pneumoniae and C. trachomatis, which were isolated from birds, mammals and humans in Japan, U.S.A., Canada and Taiwan, from 1938 to 1987. Immunological reactivity of budgerigar-derived strains to the monoclonal antibodies was different from that of pigeon-derived strains. Imported bird-derived strains were distinguishable from domestic bird-derived strains by the reactivity to the monoclonal antibodies. A close relationship between the subtypes and geographic origins was indicated on budgerigar-derived strains. On the contrary, various reactivity patterns were shown in pigeon-derived strains isolated in a narrow area. The monoclonal antibodies established in the present work may be useful probes for ecological study of avian C. psittaci.     

Bacterial black spot caused by <Emphasis Type="Italic">Burkholderia andropogonis</Emphasis> on <Emphasis Type="Italic">Odontoglossum</Emphasis> and intergeneric hybrid orchids

A new bacterial black spot disease was observed on Odontoglossum, Odontioda, Odontocidium, and Vuylstekeara orchids in Japan. Typical symptoms on the leaves were dark or black spots (or both) with a yellow halo. The causal agent was identified as Burkholderia andropogonis (Smith 1911) Gillis, Van Van, Bardin, Goor, Hebbar, Willems, Segers, Kersters, Heulin and Fernandez 1995. The isolates were pathogenic on four original host orchids, Phalaenopsis orchid, and tulip; they were not pathogenic on white clover or corn after needle stab inoculation. An antibiotic bactericide (oxytetracycline/streptomycin mixture WP) was most effective for controlling the disease.     

Incidence of silent ovulation in dairy cows during post partum period

The present study was undertaken to show the incidence of silent ovulation in high producing dairy cows, by monitoring ovarian cyclicity based on practical milk progesterone assay which was established in this study. The direct milk progesterone enzyme linked immunosorbent assay (ELISA) was developed using anti-progesterone-3(E)-carboxymethyloxime-BSA antibody for the antibody and horse radish peroxidase labeled progesterone for tracer. High sensitivity (26 pg/mL, 0.65 pg/well), high recovery rates (83-97%), high reproducibility (CV of intra-assay 4.8-11.5%; CV of inter-assay 14.3-19.1%) and high correlation between milk progesterone concentrations measured by the direct ELISA and the values obtained by the ELISA after extraction proved the reliability of the assay. In second experiment the incidence of silent ovulation was investigated based on the milk progesterone concentrations in 32 dairy cows within 70 days post partum. The incidences of silent ovulation at the first, second, third and fourth ovulation post partum were 83, 46, 13 and 0%, respectively. Most commonly observed patterns of sequential occurrence of silent ovulation in cows ovulating 2, 3 or 4 times within 70 days post partum were silent-estrus (50%), silent-silent-estrus (60%) and silent-silent-estrus-estrus (67%), respectively. These results suggest that the present ELISA is a reliable and practically applicable method for determination of progesterone in milk and that high producing dairy cows show a high incidence of silent ovulation at the first post partum ovulation as well as the second ovulation, which then decreased with the increased frequency of ovulation.     

A novel repeated sequence located on the bovine Y chromosome: its application to rapid and precise embryo sexing by PCR

A novel repeated sequence specific to male cattle was identified and named S4. S4 is a highly repetitive sequence and is a 1.5 kb repeating unit that contains various internal repeated sequences. FISH analysis showed that S4 is localized on the whole long arm and the proximal region of the short arm of the Y chromosome. We found that a PCR primer set for S4 amplified a male-specific 178 bp product in addition to a 145 bp product common to both male and female cells. Although the origin of the 145 bp product is unknown, it acts as a positive internal control in practical embryo sexing. Due to the high copy number of S4, PCR required only 0.5 pg purified DNA for accurate amplification. This made it possible to reduce the amount of biopsy sample required for embryo sexing and thus result in less damage to embryos manipulated. These studies indicate that embryo sexing based on the S4 sequence is accurate and sensitive.     

Chicken peripheral blood CD3+CD4-CD8- cells are regulated by endocrine and nerve systems

The existence of CD3(+)CD4(-)CD8(-) T cells in thymus and spleen has already been known. However, because of the presence of large amounts of thrombocytes in peripheral blood (PB), the proportion of CD3(+)CD4(-)CD8(-) T cells in PB has yet to be investigated. Therefore, the proportion of peripheral T cell-subsets was investigated in 6-week-old chickens. The percentage of CD3(+) cells, CD4(+) cells, CD8 alpha(+) cells, CD8 beta(+), and CD3(+)CD4(-)CD8(-) cells was 76%, 41%, 14%, 5%, and 15%, respectively. The proportion of CD3(+)CD4(-)CD8(-) cells in PB increased during egg-laying periods and in chickens treated with an analog of estrogen, while it decreased with age and in response to restraint stress. All of the CD3(+)CD4(-)CD8(-) cells expressed TCR1, and did not have NK activity. CD3(+)CD4(-)CD8(-) cells represent about 60% of peripheral TCR1(+) cells. These findings indicate that the proportion of CD3(+)CD4(-)CD8(-) cells is regulated by the endocrine and nerve systems.