Immunohistochemical features of canine ovarian papillary adenocarcinoma and utility of cell block technique for detecting neoplastic cells in body cavity effusions

Dogs with ovarian papillary adenocarcinoma occasionally present with ascites and/or pleural effusion. These aspirated fluids often contain a large number of cells, and distinction between neoplastic cells and activated mesothelial cells can be difficult. In this study, 7 cases of canine ovarian papillary adenocarcinoma, including 3 with ascites and pleural effusion, were immunohistochemically examined. Ovarian tumor cells were positive for cytokeratin CAM5.2 (CAM5.2), Wilms’ tumor 1 (WT-1) and progesterone receptor (PR) in all 7 cases. A metastatic lesion of the mediastinum in one case was also positive for CAM5.2, WT-1 and PR. Immunohistochemistry on cell blocks obtained from ascites and/or pleural effusion of 2 cases revealed the presence of PR-positive epithelial cells. Whereas, activated mesothelial cells in ascites or pleural effusion collected from dogs without neoplastic lesions were negative for PR. In addition, surface epithelium and subsurface epithelial structures (SES) of normal canine ovaries, that are considered to be the cell of origin for ovarian papillary adenocarcinoma, were also positive for CAM5.2, WT-1 and PR. These results indicate that, together with CAM5.2, WT-1 and PR is a useful diagnostic marker for canine ovarian papillary adenocarcinoma. Expression of PR may be associated with progesterone-dependent nature of canine ovarian papillary adenocarcinoma.     

有机肥无机肥配施对稻田氨挥发和水稻产量的影响

在南方红壤区双季稻田进行田间试验,研究等氮、磷、钾量条件下,有机无机肥配施对稻田氨挥发及水稻产量的影响。结果表明,有机无机肥配合施用能显著地降低稻田氨挥发,减少氮素损失,提高氮肥利用率。单施化肥(尿素),其氨挥发损失达37.8%,而单施有机肥和有机无机肥各半配合施用,氨挥发损失分别为0.7％-1.0％和7.2％-18.2％。田间氨挥发持续的时间,早稻约在施肥后20d,晚稻为9-10d。虽然有机无机肥各半配合施用的水稻产量与单施化肥的相近,均比对照提高约70％,但前者的氮损失少,其氮肥利用率为34.9％,高于化肥处理(33.2％)和有机肥处理(28.0%)。有机无机肥配合施用对提高水稻产量和降低氮肥环境负效应的综合效应最佳。     

Automated sampling system for long-term monitoring of nitrous oxide and methane fluxes from soils

We describe an automated gas sampling system for monitoring trace gas fluxes from soils. The sampling system allows automated collection of gas samples in glass vials using a syringe pump connected to an automated static chamber installed in the field. The gas samples are transferred to a laboratory and then analyzed using a gas chromatography system. Comparisons between manual and automated sampling of standard gases showed good agreement ( r ² = 0.99996 for N₂O, r ² = 0.999 for CH₄ and r ² = 0.998 for CO₂). In a field test, replicated flux measurements using two chambers generally showed good agreement. The sampling system allows frequent and long-term monitoring of fluxes under a wide range of weather conditions (tested temperatures ranged from –6.5 to 40°C; 127 mm day⁻¹ max precipitation). The major advantages of the system are its increased portability, ease of operation and cost effectiveness compared with on-line automated sampling/analytical systems.     

Nitrification, ammonia-oxidizing communities, and N2O and CH4 fluxes in an imperfectly drained agricultural field fertilized with coated urea with and without dicyandiamide

Agricultural soil is a major source of nitrous oxide (N₂O), and the application of nitrogen and soil drainage are important factors affecting N₂O emissions. This study tested the use of polymer-coated urea (PCU) and polymer-coated urea with the nitrification inhibitor dicyandiamide (PCUD) as potential mitigation options for N₂O emissions in an imperfectly drained, upland converted paddy field. Fluxes of N₂O and methane (CH₄), ammonia oxidation potential, and ammonia-oxidizing bacteria (AOB) and ammonia-oxidizing archaea (AOA) abundances were monitored after the application of PCU, PCUD, and urea to upland soil. The results showed that urea application increased the ammonia oxidation potential and AOB and AOA abundances; however, the increase rate of AOB (4.6 times) was much greater than that of AOA (1.8 times). These results suggested that both AOB and AOA contributed to ammonia oxidation after fertilizer application, but the response of AOB was greater than AOA. Although PCU and PCUD had lower ammonia oxidation potential compared to urea treatment, they were not effective in reducing N₂O emissions. Large episodic N₂O emissions (up to 1.59 kg N ha^?1 day^?1) were observed following heavy rainfall 2 months after basal fertilizer application. The episodic N₂O emissions accounted for 55–80 % of total N₂O emissions over the entire monitoring period. The episodic N₂O emissions following heavy rainfall would be a major source of N₂O in poorly drained agricultural fields. Cumulative CH₄ emissions ranged from ?0.017 to ?0.07 kg CH₄ ha^?1, and fertilizer and nitrification inhibitor application did not affect CH₄ oxidation.     

Isolation of Molecular Markers Linked to the <Emphasis Type="Italic">Cry</Emphasis> Locus Conferring Resistance to <Emphasis Type="Italic">Cucumber mosaic cucumovirus </Emphasis>Infection in Cowpea

We previously demonstrated that cowpea [Vigna unguiculata (L.) Walp.] cultivar Kurodane-Sanjaku contains the Cry gene, which confers resistance against Cucumber mosaic cucumovirus infection. In this paper, randomly amplified polymorphic DNA (RAPD) analysis was carried out to tag the Cry locus. Bulked segregant analysis for RAPD resulted in many polymorphisms in amplified DNA patterns. Candidates were further screened using parental and/or F₂ cowpea DNAs. As a result, we obtained three RAPD markers, D13/E14-350, WA3-850 and OPE3-500, flanking the Cry locus. In addition, we amplified cowpea sequences coding for the putative nucleotide-binding site (NBS). Degenerate primers based on NBS sequences of tobacco N and Arabidopsis RPS2 disease resistance genes were used for polymerase chain reaction, and resultant products were cloned and sequenced. Among eight independent clones, cowpea resistance gene analog (CRGA) 5 showed a distinct polymorphism when used as a probe for restriction fragment length polymorphism analysis against the susceptible cowpea cultivar PI 189375 and a near-isogenic line for the Cry. Linkage analyses of these molecular markers showed that genetic distances of CRGA5, D13/E14-350, WA3-850 and OPE3-500 to the Cry locus were 0.7, 5.2, 11.5 and 24.5 cM, respectively. Received 16 December 1999/ Accepted in revised form 22 March 2000     

Dystrophin-deficient muscular dystrophy in a Toy Poodle with a single base pair insertion in exon 45 of the Duchenne muscular dystrophy gene

A 10-month-old, intact male Toy Poodle was referred for a postural abnormality. Blood biochemical tests revealed a marked increase in plasma creatine phosphokinase (CPK) concentration. The isoenzyme test showed that 99% of serum CPK consisted of CPK-MM. Histopathological evaluation of muscle biopsy samples confirmed scattered degeneration and necrosis of myofibers. Immunohistochemistry for dystrophin showed an absence of staining in muscle cells. Based on these findings, the dog was diagnosed with dystrophin-deficient muscular dystrophy. Whole genome sequencing using genomic DNA extracted from blood revealed a single base pair insertion in exon 45 of the Duchenne muscular dystrophy (DMD) gene. This is the first report on muscular dystrophy in Toy Poodles and identified a novel mutation in the DMD gene.     

A case of feline temporal lobe epilepsy with hippocampal sclerosis and dentate gyrus malformation

A two-months-old, male, mixed breed cat presented with epileptic seizures. The cat was diagnosed with drug-resistant epilepsy, and died at 3-years of age. No gross lesion was found at necropsy. Histopathologically, the dentate gyrus granule cell layer of the hippocampus was irregularly arranged. Granule cells were dispersed and ectopic cells were sporadically observed in the molecular layer. The granule cells had an enlarged cytoplasm and swollen nucleus. Immunohistochemistry for NeuN and GFAP confirmed severe neuronal loss and mild gliosis in CA1. Binucleation and ischemic change were observed in the remaining pyramidal cells. This report describes a case of feline temporal lobe epilepsy and hippocampal sclerosis associated with dentate gyrus malformation.     

Methanogenic Characteristics of Flooded Rice Soils in Response to Glucose Amendment

Extract

It was reported that flooded rice soil was an important source of atmospheric CH₄ (Cicerone and Shetter 1981; Sciler et al. 1984). In flooded rice soils, CH₄ is produced under strict anaerobic conditions by methanogenic bacteria (Jones et al. 1987; Jones 1991). Rice plants carry the produced CH₄ from anoxic sediment and release it to the atmosphere (Cicerone and Shetter 1981; Nouchi et al. 1990). More than 90% of emitted CH₄ is released through rice plants and the emission pattern usually shows large seasonal variations (Cicerone et al. 1983; Sciler et al. 1984; Inubushi et al. 1989). Since the number of methanogens did not change throughout the rice growth season (Schutz et al. 1989; Mayer and Conrad 1990), seasonal variations of CH₄ emission could be ascribed to the activities of methanogens and the amount of available substrates. However, the changes in the methanogenic activities in response to available substrates especially in relation to the growth period of rice plants are poorly documented.     

Dynamics of microbial biomass nitrogen as influenced by organic matter application in paddy fields

The effects of annual application of rice straw or cow manure compost for 17–20 y on the dynamics of fertilizer N and soil organic N in Gley paddy fields were investigated by using the ¹⁵N tracer technique during the rice cropping season. The chloroform fumigation-extraction method was evaluated to determine the properties of soil microbial biomass under submerged field conditions at the tillering stage before mid-summer drainage, with special reference to the fate of applied NH₄ ⁺-¹⁵N.

The transfer ratios from applied NH₄ ⁺-¹⁵N to immobilized N in soil and to uptake N by rice during given periods varied with the rice growth stages and were affected by organic matter application. The accumulated amounts of netmineralized soil organic N (net-M_j ), immobilized N (I_j ), and denitrified N (D_j ) during the cropping season were estimated to be 14.0–22.5, 6.3–11.2, and 3.4–5.3 g N m^-2, respectively. Values of net-M_j and I_j were larger in the following order: cow manure compost plot > rice straw plot > plot without organic matter application, and their larger increase by the application of cow manure compost contributed to a decrease of the D_j values, as compared with rice straw application.

Values of E _N extra extractable soil total N after fumigation, increased following organic matter application, ranging from 2.1 to 5.4 g N m^-2. Small residual ratios of applied ¹⁵N in the fraction E _N at the end of the given period indicated that re-mineralization of newly-assimilated ¹⁵N through the easily decomposable fraction of microbial biomass had almost ended. Thus, the applicability to paddy field soils of the chloroform fumigation-extraction method was confirmed.