Changes in the soil C and N contents,C decomposition and N mineralization potentials in a rice paddy after long-term application of inorganic fertilizers and organic matter

A long-term experiment on combined inorganic fertilizers and organic matter in paddy rice (Oryza sativa L.) cultivation began in May 1982 in Yamagata, northeastern Japan. In 2012, after the 31^st harvest, soil samples were collected from five fertilizer treatments [(1) PK, (2) NPK, (3) NPK + 6 Mg ha^?1 rice straw (RS), (4) NPK + 10 Mg ha^?1 rice straw compost (CM1), and (5) NPK + 30 Mg ha^?1 rice straw compost (CM3)], at five soil depths (0–5, 5–10, 10–15, 15–20 and 20–25 cm), to assess the changes in soil organic carbon (SOC) content and carbon (C) decomposition potential, total nitrogen (TN) content and nitrogen (N) mineralization potential resulting from long-term organic matter addition. The C decomposition potential was assessed based on the methane (CH₄) and carbon dioxide (CO₂) produced, while the N mineralization potential was determined from the potassium chloride (KCl)-extractable ammonium-nitrogen (NH₄⁺-N), after 2, 4, 6 and 8 weeks of anaerobic incubation at 30°C in the laboratory. Compared to NPK treatment, SOC in the total 0–25 cm layer increased by 67.3, 21.0 and10.8%, and TN increased by 64.2, 19.7 and 10.6%, in CM3, RS and CM1, respectively, and SOC and TN showed a slight reduction in the PK treatment by 5.2 and 5.7%, respectively. Applying rice straw compost (10 Mg ha^?1) instead of rice straw (6 Mg ha^?1) to rice paddies reduced methane production by about 19% after the soils were measured under 8 weeks of anaerobic incubation at 30°C. Soil carbon decomposition potential (Co) and nitrogen mineralization potential (No) were highly correlated with the SOC and TN contents. The mean ratio of Co/No was 4.49, lower than the mean ratio of SOC/TN (13.49) for all treatments, which indicated that the easily decomposed organic matter was from soil microbial biomass and soil proteins.     

Rapid identification of CCA-treated wood using laser-induced breakdown spectroscopy

As chromated copper arsenate (CCA) contains copper, chromium and arsenic, waste CCA-treated wood must be separated from other treated wood because of environmental pollution by chromium and arsenic when it is incinerated and the regulation. Therefore, a method to identify CCA-treated wood was developed using laser-induced breakdown spectroscopy (LIBS). Using the LIBS apparatus assembled in our laboratory, plasma on a wood surface was generated by a 4?ns pulse of 1064?nm (55?mJ/mm²) emitted from Nd:YAG laser. Fluorescence from the plasma was collected by an ellipsoidal mirror and analyzed by a spectrometer in the range of 190–300?nm. The results showed that the 228.7?nm line from As and 267.6?nm line from Cr were useful for the identification of CCA-treated wood. As the discrimination capacity was confirmed by the elemental composition analysis by X-ray fluorescence, it was concluded that LIBS can specifically identify CCA-treated wood.     

Phylogenetic relationships of three species within the family Heligmonellidae (Nematoda; Heligmosomoidea) from Japanese rodents and a lagomorph based on the sequences of ribosomal DNA internal transcribed spacers, ITS-1 and ITS-2

Nematodes of the family Heligmonellidae (Heligmosomoidea; Trichostrongylina) reside in the digestive tracts of rodents and lagomorphs. Although this family contains large numbers of genera and species, genetic information on the Heligmonellidae is very limited. We collected and isolated adult worms of three species in Japan that belong to the family Heligmonellidae, namely Heligmonoides speciosus (Konno, 1963) Durette-Desset, 1970 (Hs) from Apodemus argenteus, Orientostrongylus ezoensis Tada, 1975 (Oe) from Rattus norvegicus and Lagostrongylus leporis (Schulz, 1931) (Ll) from Pentalagus furnessi, and sequenced the entire internal transcribed spacer regions, ITS-1 and ITS-2 of ribosomal DNA. ITS-1 of Hs, Oe and Ll was 426, 468 and 449 bp in length, and had a G+C content of about 41, 41 and 37 %, respectively. ITS-2 of Hs, Oe and Ll was 297, 319 and 276 bp in length and had a G+C content of about 38, 40 and 28%, respectively. The data of Hs, Oe and Ll were compared with those of two other known species within the family Heligmonellidae, Calorinensis minutus (Dujardin, 1845) (Cm) and Nippostrogylus brasiliensis (Travassos, 1914) (Nb), and with those of two species of Heligmosomidae (Heligmosomoidea), Heligmosomoides polygyrus bakeri and Ohbayashinema erbaevae. Phylogenetic analysis placed Hs, Oe and Ll in the same clade with Cm and Nb, forming a Heligmonellidae branch in both ITS-1 and ITS-2, separate from the Heligmosomoidea branch. These results demonstrated that the ITS-1 and ITS-2 sequences are useful for differentiating the Heligmonellidae nematode species. This study is the first to describe the ITS-1 and ITS-2 sequences of Hs, Oe and Ll.     

Hypoxanthine enhances the cured meat taste

We evaluated the enhancement of cured meat taste during maturation by sensory analysis. We focused on the heat‐stable sarcoplasmic fraction (HSSF) to identify the factors related to cured meat taste. Because the dry matter of HSSF contained more than 30% nitrogen, nitrogen compounds such as free amino acids, small peptides and adenosine triphosphate‐related compounds seemed to be the important components of HSSF. The samples cured with HSSF for 2 h exhibited the same taste profile as ones cured without HSSF for 168 h. Therefore, the changes in the amount and fractions of nitrogen compounds were examined in HSSF during incubation from 0 to 168 h. The concentration of hypoxanthine (Hx) gradually increased, while inosine‐5′‐monophosphate decreased during the incubation. The samples cured with pickles containing various concentrations of Hx were subjected to sensory analysis. The addition of Hx, in a dose‐dependent fashion, enhanced cured meat taste by maturation for 2 h. It was concluded that Hx is essential for the enhancement of cured meat taste.     

Cytosolic 20 alpha-hydroxysteroid dehydrogenase activity in spontaneous neoplasms in the dog and cat.

Steroid-producing tissues such as ovary, placenta, testis and adrenal gland and non-steroid-producing cells including lymphocytes are known to contain 20 alpha-hydroxysteroid dehydrogenase (20 alpha-HSD). In the present study, we measured and partially characterized the 20 alpha-HSD in 11 neoplastic tissues surgically removed from dogs and a cat. The tissues were pathologically classified as 4 benign and 7 malignant. The enzyme activities in the cytoplasmic fraction were positive in 6 of the 7 malignant tissues and 2 of the 4 benign ones. Following DEAE chromatography analysis of 2 malignant tumour samples, multiple forms of enzyme activities with different electric charges were detected. Furthermore, 20 alpha-HSD activity in the cytosol fraction from malignant tumours was increased dramatically by passage through the DEAE column, suggesting that the enzyme is present in the cytosol as an inactive or sequestered form.     

Detection of radiation-induced apoptosis using the comet assay

The electrophoresis pattern of apoptotic cells detected by the comet assay has a characteristic small head and spread tail. This image has been referred to as an apoptotic comet, but it has not been previously proven to be apoptotic cells by any direct method. In order to identify this image obtained by the comet assay as corresponding to an apoptotic cell, the frequency of appearance of apoptosis was examined using CHO-K1 and L5178Y cells which were exposed to gamma irradiation. As a method for detecting apoptosis, the terminal deoxynucleotidyl transferase mediated dUTP nick end labeling (TUNEL) assay was used. When the frequency of appearance of apoptotic cells following gamma irradiation was observed over a period of time, there was a significant increase in appearance of apoptosis when using the TUNEL assay. However, there was only a slight increase when using the comet assay. In order to verify the low frequency of appearance of apoptosis when using the comet assay, we attempted to use the TUNEL assay to stain the apoptotic comets detected in the comet assay. The apoptotic comets were TUNEL positive and the normal comets were TUNEL negative. This indicates that the apoptotic comets were formed from DNA fragments with 3'-hydroxy ends that are generated as cells undergo apoptosis. Therefore, it was understood that the characteristic pattern of apoptotic comets detected by the comet assay corresponds to cells undergoing apoptosis.     

Passive impregnation of liquid in impermeable lumber incised by laser

Square lumber specimens of laser-incised Douglas fir (Pseudotsuga menziesii Franco) were treated with steam before dipping. Two types of steam (saturated steam and superheated steam), three steam-injection times (5, 10, and 20 min), four different time intervals (moving time) between steam treatment and dipping (immediate, 3, 10, and 30 min), and four different dipping times (0.5, 1, 3, and 12 h) were used in the study. The maximum absorption was 480 kg/m³ when saturated steam was injected for 20 min and the specimen was immediately dipped into liquid for 12 h. Samples treated with this condition not only absorbed the maximum amount of liquid but also penetrated over 83.4% and 87.3% of the total area along and across the grain, respectively. The optimum conditions were then applied to laser-incised sugi (Cryptomeria japonica D. Don) and Japanese larch (Larix leptolepis Gordon) where the absorption of liquid was 415 and 187 kg/m³, respectively. It was shown that initial moisture content below the fiber saturation point was good for passive impregnation. The absorption of liquid and its distribution in wood indicates that it can be a good preservative treatment method for impermeable woods.     

Development of an in vitro model for the analysis of bovine endometrium using simple techniques

This study aimed to develop an in vitro model for the analysis of the bovine endometrium. Immunofluorescent staining revealed that the hetero‐spheroids and the cultured explants showed almost similar structure in the localization of bovine endometrial epithelial cells and endometrial stromal cells, except the glandular‐like structure of the epithelial cells inside the explants. Gelatin zymography revealed that the hetero‐spheroids did not express matrix metalloproteinases (MMPs) after 4 days of culture, but strong MMP expressions were observed in the cultured explants until 7 days of culture. Additionally, expression of progesterone receptor (PR), estrogen receptor alpha (ERα), type I interferon receptor 1 (IFNAR1) and 2 (IFNAR2) messenger RNA was observed both in the homo‐ and hetero‐spheroids. The expression of oxytocin receptor (OTR) mRNA in E2 and E2+P4 (1,3,5(10)‐Estratrien‐3, 17β‐diol + 4‐Pregnen‐3, 20‐dinone) treated groups were significantly (P < 0.05) higher than that of the control group of spheroids. In case of cultured explants, the expression of PR and OTR mRNA were significantly (P < 0.05) higher in E2 treated groups compared to the control groups. Hepatocyte growth factor (HGF) mRNA expression was also higher in P4 treated groups at 10 days in culture (P < 0.05). In a nutshell, the in vitro model developed in this study for the analysis of the endometrium may provide a new platform for extensive research on bovine endometrial function.     

The importance of subfragment 2 and C‐terminus of myosin heavy chain for thick filament assembly in skeletal muscle cells

In skeletal muscle cells, myofibrillar proteins are highly organized into sarcomeres in which thick filaments interdigitate with thin filaments to generate contractile force. The size of thick filaments, which consist mainly of myosin molecules, is strictly controlled. However, little is known about the mechanisms by which myosin molecules assemble into thick filaments. Here, we assessed the ability of each domain of myosin heavy chain (Myh) to form thick filaments. We showed that exogenously expressed subfragment 2 (S2) + light meromyosin (LMM) of Myh was efficiently incorporated into thick filaments in muscle cells, although neither solely expressed S2 nor LMM targeted to thick filaments properly. In nonmuscle COS7 cells, S2+LMM formed more enlarged filaments/speckles than LMM. These results suggest that Myh filament formation is induced by S2 accompanying LMM. We further examined the effects of Myh C‐terminus on thick filament assembly. C‐terminal deletion mutants were incorporated not into entire thick filaments but rather into restricted regions of thick filaments. Our findings suggest that the elongation of myosin filaments to form thick filaments is regulated by S2 as well as C‐terminus of LMM.     

Chondrocyte apoptosis and decrease of glycosaminoglycan in cranial cruciate ligament insertion after resection in rabbits

We examined time-dependent histological changes of the calcified fibrocartilage area in a tibial cranial cruciate ligament (CCL) insertion after ligament resection in rabbits. The animals were divided into two groups: those undergoing CCL substance resection in the right stifle (resected group) and those receiving the same operation without CCL resection in the left stifle (sham operated group). Five animals were euthanized with deep anaesthesia at four time periods (1, 2, 4 and 6 weeks), and Haematoxylin-eosin and Safranin-O stainings and terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate-biotin nick end labeling (TUNEL) staining were performed. The average percentage of TUNEL-positive chondrocytes and the average thickness of the glycosaminoglycan (GAG)-stained area in the calcified fibrocartilage area were measured. Two and 4 weeks after the surgery, the average percentages of TUNEL-positive chondrocytes in the resected group (23.8 +/- 10.3% and 15.9 +/- 6.7%, respectively) were significantly higher than those in the sham operated group (8.9 +/- 3.8% and 7.4 +/- 1.6%, P<0.05, respectively). Six weeks after the surgery, the average thickness of the GAG-stained area in the resected group (7.7 +/- 13. 5 microm) was significantly smaller than that in the sham operated group (69.4 +/- 39.9 microm, P<0.05). Our results suggest that the average percentage of TUNEL-positive chondrocytes became a peak in 2 weeks and that histological changes occurred in 6 weeks. The chondrocyte apoptosis can induce decrease of GAG-stained area after resection of CCL. Therefore, chondrocyte apoptosis in the calcified cartilage area in the CCL tibial insertion might lead to histological changes.