Histochemical and immunohistochemical characterization of chordoma in ferrets

Chordomas of the tip of the tail in 6 ferrets were examined using histopathological, histochemical and immunohistochemical procedures. Histopathologically, round neoplastic cells containing numerous cytoplasmic vacuoles of varying sizes, categorized as “physaliphorous cells”, were observed in the amorphous eosinophilic or pale basophilic myxoid stroma. Physaliphorous cells were arranged in lobules and in a “chordoid” or “cobblestone” manner. The neoplasms were diagnosed as benign chordoma without local invasion and metastasis. Histochemically, the cytoplasm of small neoplastic cells was positive for periodic acid-Schiff stain and alcian blue (AB) pH 2.5 and pH 1.0 stains, but negative for hyaluronidase digestion-AB pH 2.5 stain. All neoplastic cells were strongly stained with colloidal ion, negative for high iron diamine AB pH 2.5 and toluidine blue pH 2.5 stains, and positive for Mayer’s mucicarmine stain. Immunohistochemistry using antibodies directed against low-molecular-weight cytokeratins (CK18, CK19 and CK20), vimentin and mucin core protein (MUC5AC) revealed that neoplastic cells had both epithelial and mesenchymal elements. The expression of low-molecular-weight cytokeratins suggests that neoplastic cells acquired the properties of glandular epithelial cells and produced epithelial mucus. Furthermore, the expression of cytokeratins, vimentin, S100 protein, brachyury and epithelial membrane antigen indicates that the neoplasms were equivalent to the classic type of human chordoma. Therefore, immunohistochemistry using these antibodies can be useful for the characterization of ferret chordoma.     

Cesium radioactivity in peripheral blood is linearly correlated to that in skeletal muscle: Analyses of cattle within the evacuation zone of the Fukushima Daiichi Nuclear Power Plant

The accident at the Fukushima Daiichi Nuclear Power Plant (FNPP) released a large amount of radioactive substances into the environment. Furthermore, beef contaminated with radioactive cesium above the 500 Bq/kg safety standard was circulated in the food chain in 2011. Japanese consumers remain concerned about the safety of radioactively contaminated food. In our previous study, we detected a linear correlation between radioactive cesium (¹³⁷Cs) activity in blood and muscle around 500 to 2500 Bq/kg in cattle. However, it was unclear whether the correlation was maintained at a lower radioactivity close to the current safety standard of 100 Bq/kg. In this study, we evaluated 17 cattle in the FNPP evacuation zone that had a ¹³⁷Cs blood level less than 10 Bq/kg. The results showed a linear correlation between blood ¹³⁷Cs and muscle ¹³⁷Cs (Y = 28.0X, R² = 0.590) at low radioactivity concentration, indicating that cesium radioactivity in the muscle can be estimated from blood radioactivity. This technique would be useful in detecting high‐risk cattle before they enter the market, and will contribute to food safety.     

Fractal analysis of ice crystals in frozen food

Fractal analysis of the morphology of ice crystal particles in frozen food was attempted for frozen soybean curd (tofu). From a microscopic image analysis of the ice crystal particles, it was found that the perimeter of the ice crystal particles could be recognized as a fractal. Effects of the storage time and storage temperature on the fractal dimension (d(p)) of the perimeter of the ice crystal particles were also investigated. As storage time was increased, the d(p) value tended to decrease. When the storage temperature was increased, the value of d(p) decreased more rapidly. The changes corresponded to the visual observation of the shape change for the ice crystal particles during storage reported by many researchers, suggesting that the fractal dimension d(p) could be used as a quantitative indicator reflecting the surface roughness of ice crystal particles.     

Microbial metabolism in reduction process of paddy soils (Part 2)

The previous experiences^1-3) with incubation experiments of the soils under submargd condition indicated that the disappearance of molecular oxygen and nitrates occurred shortly after sub-emerging, and then the reduction of ferric iron proceeded, being followed by the reduction of sulfates and the formation of methane which nre caused by the activities of strict anaerobes. It was suggested from the above fact that the reduction process in the submarged soils was affected mainly by two factors: namely, the amount of reducible ferric iron and the soil organic matters which are avaiable as substrate for microbial metabolism.     

Effectiveness of tetrazolium salts in microbial ecological studies in submerged soil

Several trials to obtain information on the living microorganisms at micro-sites of the submerged soil by the novel utilization of the tetrazolium salts were described. Results are summarized as follows:

(1) The air-dried soil was incubated with TIC (triphenyl tetrazolium chloride) under submerged condition according to the modified double layer plate method. The red formazan (the reduced product of TIC) was precipitated at the biologically active sites by the dehy. drogenase activity. Using a binocular microscope, most of the biologically active sites were found to be localized on and around the organic debris.

(2) The amount of nitrogen mineralized by incubating the soil under submerged condi. tion correlated well not only with the amount of formazan formed biologically in the submerged soil but also with the amount of formazan formed non-biologically by heating the soil with TIC in Ba(OH). solution. In addition, the pattern of precipitation of the nonbiologically formed formazan was very similar to that of the biologically formed formazan. Accordingly, the substances which reduced TIC non-biologically were considered to be closely associated with the substrates for the microorganisms.

(3) The plant debris stained with the red formazan could be mounted on a slide glass and observed at high magnification for the detailed study on the process of decomposition. Addition of lactophenol cotton blue to the plant debris could visualize the microorganisms responsible for the decomposition of the plant debris.

(4) NTB (nitro tetrazolium blue) was the most suitable for staining individual microorganisms growing on cellulosic materials or plant debris. The double staining with NTB and rosebengal made the discrimination between dead or resting cells and viable ones possible when this technique was applied to the decomposed filter paper.     

Factors Influencing the Determination of Microbial Biomass-Nitrogen in Submerged Soil

In a previous paper (1), a modification of the chloroform fumigation method (2) was proposed for estimating microbial biomass-nitrogen (N) in submerged soil. It was found that a large part of the living bacteria in the submerged soil was killed within about 12 h of the fumigation and was actively mineralized by subsequent anaerobic incubation.

In this paper, the following factors which influence the estimated amount of microbial biomass-N were examined: a) inoculation to fumigated soil, b) duration of anaerobic incubation, c) storage and sieving pretreatments of soil, and d) effect of fumigation on soil organic matter.     

Expression analysis of defense-related genes in wild kiwifruit (Actinidia rufa) tolerant to bacterial canker

Journal of General Plant Pathology - Kiwifruit bacterial canker, which is caused by Pseudomonas syringae pv. actinidiae biovar 3 (Psa3), is found throughout kiwifruit-growing countries. Here, we...     

Comparison of acute inhalation toxicity of sulfuric acid by the inhalation and intratracheal instillation methods

Recently, intratracheal instillation has been focused on as a simple, low-cost alternative to the inhalation method. In this study, intratracheal instillation of sulfuric acid, a typical acidic compound, was performed to compare the acute toxicity of acidic compounds that could cause damage to the respiratory system between intratracheal instillation and inhalation. Sulfuric acid was administered to male rats at doses of 0.7, 2, 7, 20, and 60 mg/kg by dividing the total dose into four doses. General condition and body weight were examined up to 14 days after administration, and macropathological and histopathological examinations were performed. The half-lethal dose was then estimated. All animals administered 20 and 60 mg/kg sulfuric acid and one animal administered 2 mg/kg sulfuric acid died within 4 h after administration. No abnormalities were observed in other animals. At 20 and 60 mg/kg, multiple red foci or diffuse red areas were macroscopically observed in the lungs. In these lesions, histopathologically, clefts between the mucosal epithelium and basement membrane and necrosis of the alveolar epithelium were observed. Deaths in these groups may have resulted from lung injury. No notable changes were observed in other animals. Therefore, the half-lethal dose of sulfuric acid by intratracheal instillation was estimated as 7–20 mg/kg. The acute toxicity by intratracheal instillation was evaluated with two-fold sensitivity since the exposure at the half-lethal sulfuric acid concentration in inhalation studies was calculated as 43.2 mg/kg.     

Purification and characterization of polyphenol oxidase from cauliflower (Brassica oleracea L.)

Polyphenol oxidase (PPO) of cauliflower was purified to 282-fold with a recovery rate of 8.1%, using phloroglucinol as a substrate. The enzyme appeared as a single band on sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The estimated molecular weight of the enzyme was 60 and 54 kDa by SDS-PAGE and gel filtration, respectively. The purified enzyme, called phloroglucinol oxidase (PhO), oxidized phloroglucinol (K(m) = 3.3 mM) and phloroglucinolcarboxylic acid. The enzyme also had peroxidase (POD) activity. At the final step, the activity of purified cauliflower POD was 110-fold with a recovery rate of 3.2%. The PhO and POD showed the highest activity at pH 8.0 and 4.0 and were stable in the pH range of 3.0-11.0 and 5.0-8.0 at 5 °C for 20 h, respectively. The optimum temperature was 55 °C for PhO and 20 °C for POD. The most effective inhibitor for PhO was sodium diethyldithiocarbamate at 10 mM (IC(50) = 0.64 and K(i) = 0.15 mM), and the most effective inhibitor for POD was potassium cyanide at 1.0 mM (IC(50) = 0.03 and K(i) = 29 μM).     

Transactivational property of chemicals via medaka glucocorticoid receptor 1b using a stable reporter gene assay

The transactivational property of natural and synthetic chemicals via medaka GR1b was investigated after development of a stable cell line for the reporter gene assay. In our study, cortisol was the most potent agonist among the natural corticoids assayed for potency [EC₅₀ (concentration of agonist provoking a response halfway between the baseline and maximum response) 68 nM] and efficacy. Three artificial corticosteroids, namely, dexamethasone [EC₅₀ 16 nM, relative agonistic activity to cortisol (RAA) 144 %], prednisolone (EC₅₀ 81 nM, RAA 116 %) and clobetasol propionate (EC₅₀ 0.10 nM, RAA 220 %), showed strong agonistic activity and were more potent than the original corticoid, F. All synthetic corticoids used in our study were full agonists. Interestingly, melengestrol acetate, a synthetic progestogen, induced luciferase activity in a dose-dependent manner. Based on its EC₅₀ value and RAA of 29 nM and 57 %, respectively, this molecule was assessed as a partial agonist. None of the other steroids and chemicals assayed in our study induced an agonistic response. In conclusion, we successfully developed a stable reporter gene assay that can be used to assess the transactivational property of glucocorticoid-like chemicals toward medaka GR1b.