Molecular cloning and characterization of chicken tumor necrosis factor (TNF)-superfamily ligands, CD30L and TNF-related apoptosis inducing ligand (TRAIL)

CD30 ligand (CD30L) and tumor necrosis factor (TNF)-related apoptosis inducing ligand (TRAIL) are members of the TNF-superfamily that have many important biological activities in cell proliferation and apoptotic death. In this study, both genes in the chicken were cloned and their expression was analyzed. Complementary DNA fragments were obtained from a suppressive subtractive hybridization library with or without lipopolysaccharide (LPS)-stimulation. Chicken CD30L consists of 1,152 base pairs (bp) with an open reading frame (ORF) of 720 bp having 36.4% identity with human CD30L, whereas chicken TRAIL is 1,134 bp long with an ORF of 912 bp having 54.4% identity with human TRAIL. Chicken CD30L was expressed at high levels in the spleen, bursa of Fabricius and in the chicken monocytic leukemia cell line, IN24. Stimulation with LPS in the spleen, bursa of Fabricius and the IN24 cell line did not affect CD30L expression. The gene expression of chicken TRAIL was essentially to the same level in all tissues examined. The time course of expression was not significantly altered by LPS-stimulation in the spleen, thymus and bursa of Fabricius, but reached a maximal level 8 hr after stimulation in the IN24 cell line. The high level expression of both genes in lymphoid organs and IN24 cell line indicates that chicken CD30L and TRAIL may also play an important role in apoptotic signal transduction and the regulation of cell proliferation in the immune system.     

Connective tissue-type mast cell leukemia in a dog

An unusual case diagnosed as connective tissue-type mast cell leukemia with marked mastocyte infiltration into visceral organs in a seven-year-old female Curly-Coated retriever is presented. Acute circulatory collapse, emesis, diarrhea, abdominal enlargement, icterus, cyanosis, dyspnea, pulmonary edema, hepatomegary, ascites, and right ventricular enlargement were observed. Hematologic and biochemical examinations revealed mast cell leukemia, mature neutrophilia, monocytosis, thrombocytopenia, hemolytic hyperbilirubinemia, hyperhistaminemia, renal and hepatic injuries. Mast cells were distributed systemically, but predominantly in the diaphragm and liver with a large mass among the serosa of ileum, cecum and colon. Mast cells were stained intensely by both safranin and berberine sulfate.     

Stabilization of atomic hydrogen in both solution and crystal at room temperature

Atomic hydrogen has been stably encapsulated in both solution and crystal at room temperature. Upon gamma-ray irradiation of [(CH(3))(3)Si](8)Si(8)O(20), which is the trimethylsilylated derivative of the silicate anion with a double four-ring (D4R) cage, electron spin resonance (ESR) spectra revealed that a single hydrogen atom is encapsulated in the center of the D4R cage and is stable for periods of many months. Attack by chemically reactive species such as oxygen was prevented by the D4R cage, but the ESR signal of the hydrogen atom was sensitive to the magnetic interaction caused by the presence of the O(2) molecule near the cage.     

Transgenic rice plants expressing human CYP1A1 remediate the triazine herbicides atrazine and simazine

The human cytochrome P450 CYP1A1 gene was introduced into rice plants (Oryza sativa cv. Nipponbare). One-month-old CYP1A1 plants grown in soil clearly showed a healthy growth and tolerance to 8.8 microM atrazine and 50 microM simazine, but nontransgenic plants were completely killed by the herbicides. Although transgenic and nontransgenic plants metabolized the two herbicides into the same sets of compounds, CYP1A1 plants metabolized atrazine and simazine more rapidly than did control plants. In small-scale experiments, residual amounts of atrazine and simazine in the culture medium of CYP1A1 plants were 43.4 and 12.3% of those in control medium; those of nontransgenic Nipponbare were 68.3 and 57.2%, respectively. When cultivated in soil with 2.95 microM atrazine and 3.15 microM simazine for 25 days, CYP1A1 plants eliminated 1.3 times more atrazine and 1.4 times more simazine from the soil than did control plants. Thus, CYP1A1 rice plants make it possible to remove atrazine and simazine more rapidly from the culture medium and soil than can nontransgenic Nipponbare.     

Phytoremediation of metolachlor by transgenic rice plants expressing human CYP2B6

We introduced the human cytochrome P450 gene CYP2B6 into rice plants (Oryza sativa L. cv. Nipponbare), and the CYP2B6-expressing rice plants became more tolerant to various herbicides than nontransgenic Nipponbare rice plants. In particular, CYP2B6 rice plants grown in soil showed tolerance to the chloroacetanilide herbicides alachlor and metolachlor. We evaluated the degradation of metolachlor by CYP2B6 rice plants to confirm the metabolic activity of the introduced CYP2B6. Although both CYP2B6 and nontransgenic Nipponbare rice plants could decrease the amount of metolachlor in plant tissue and culture medium, CYP2B6 rice plants could remove much greater amounts. In a greenhouse, the ability of CYP2B6 rice plants to remove metolachlor was confirmed in large-scale experiments, in which these plants appeared able to decrease residual quantities of metolachlor in water and soil.     

Molecular characterization of <Emphasis Type="Italic">Potato spindle tuber viroid</Emphasis> in dahlia

A new variant of Potato spindle tuber viroid (PSTVd) was detected for the first time from dahlia grown in Japan. The dahlia isolate of PSTVd formed a quasi-species and a major sequence variant consisting of 361 nucleotides in length, including five substitutions, three insertions, and one deletion, when compared to the intermediate strain from potato. In bioassays with the new isolate, Rutgers tomato developed mild stunting and leaf curling.     

A Case Study on Estimation of Dry Deposition of Sulfur and Nitrogen Compounds by Inferential Method

In order to estimate dry deposition, deposition velocity calculation and concentration measurement were carried out in Niigata, Japan. Deposition velocities of SO₂ and HNO₃ for some surfaces such as coniferous forest, deciduous forest, agricultural land, and water were calculated taking into account diurnal variations of meteorological elements using routine meteorological data. Deposition velocities of fine and coarse aerosols were also estimated respectively. Concentrations of SO₂, HNO₃, sulfate and nitrate in fine and coarse aerosols respectively were measured from July to December in 1998 using filter pack and denuder methods. The results indicate that HNO₃ dry deposition for the high aerodynamic roughness surfaces such as forests is quite large. It is suggested that sulfate and nitrate aerosols depositions as well as the gases depositions should be taken into account to estimate dry deposition of sulfur and nitrogen compounds. It is also indicated that dry depositions of sulfur and nitrogen compounds are unable to ignore compared with their wet depositions.     

Glucocorticoid sensitivity depends on expression levels of glucocorticoid receptors in canine neoplastic mast cells

Glucocorticoid (GC) administration with or without other chemotherapeutic reagents is a commonly used option in the treatment of mast cell malignancies. However, the responsiveness of mast cell tumors to GC treatment varies in individuals, and the regulatory mechanisms determining the GC sensitivity of malignant mast cells remain unclear. Since the expression of the GC receptor (GR) has been reported to be associated with GC sensitivity in human neoplastic lymphocytes, we attempted to investigate the relationship between GR levels and GC sensitivity by using neoplastic mast cells derived from canine mast cell tumors (MCTs). To elucidate the regulatory mechanisms involved in GC responsiveness, we analyzed various canine MCT cell lines and tissue samples from dogs with MCT. While the proliferation of canine MCT cells was suppressed by the addition of GC to the culture, we found that MCT cells derived from humans and rodents, as well as canine lymphoma cells, responded poorly to GC. However, there were also some variations in responsiveness to GC treatment among canine MCT cell lines used in this study. Using real-time polymerase chain reaction and Western blot analysis, we elucidated the relationship between GR expression and responsiveness to GC in canine MCT cells. Furthermore, to assess the involvement of GR expression in GC sensitivity in vivo, clinical investigations were conducted on dogs with cutaneous MCT. Written informed consent was obtained from owners, and the affected dogs were treated with prednisolone (0.5-2.0 mg kg(-1)day(-1), administered orally) 1 or 2 weeks prior to the surgical removal of the tumors. Tumor volume was measured according to WHO criteria both before and after prednisolone treatment, and the GC sensitivity of each MCT was determined on the basis of the reduction in tumor volume. Of the 15 dogs with MCT, 11 responded to treatment with prednisolone completely or partially, whereas 4 dogs showed no response. Examination of clinical samples obtained by surgical removal revealed that GR expression levels were significantly lower in GC-resistant MCT tissues than in GC-sensitive MCT tissues. Thus, these results strongly indicate that GR expression may contribute to GC sensitivity in canine MCT.