Interaction between myostatin and extracellular matrix components

Myostatin, a member of the TGF-β superfamily, is a negative regulator of skeletal muscle mass. We have recently demonstrated that decorin binds to myostatin in vitro , and that immobilized decorin within the collagen matrix prevents myostatin-mediated inhibition of myoblast proliferation. However, little is known about other ECM molecules that bind to myostatin and modulate its activity. Thus, in the present study, we investigated the interaction of several other ECM molecules with myostatin. We here show that fibromodulin, fibronectin and laminin bind to myostatin in the presence of Zn²⁺ with a dissociation constant ( K_D ) of 10⁻¹⁰∼10⁻⁸ mol/L. Fibromodulin shows the highest affinity for myostatin among them. These results suggest that these ECM molecules may modulate myostatin activity like decorin does.     

Pathogenic characters of Japanese potato strains of Ralstonia solanacearum

Ralstonia solanacearum (Rs) strains in phylotypes I and IV isolated from potato in Japan were investigated for pathogenicity on potato, tomato, eggplant, Solanum integrifolium, tobacco, groundnut, and pumpkin. The strains were divided into 17 types based on differences in their pathogenicity on the tested plants. Particularly, the pathogenicity of most phylotype I strains on eggplant was distinctly different from that of the phylotype IV strains. When nine potato varieties (included two breeding lines) were inoculated with several Rs strains, phylotype IV strains were highly virulent on the breeding lines that are regarded as resistant to phylotype I strains.     

Orally administered rutin inhibits the gene expression of Th2 cytokines in the gut and lung in aged mice

Rutin is one of the flavonoids derived from plants such as buckwheat and is well known as a powerful antioxidant. To determine whether dietary rutin could modulate mucosal immunity, we examined the gene expression of Th1/Th2 cytokines and the receptors in the gut and lung. Aged (18 months old, 18 M) C3H/HeN female mice were orally administered rutin for 10 days. The small intestine and lung were taken and analyzed by real-time PCR for gene expression. Interleukin (IL)-13 and IL-13Rα2 gene expression was significantly low (P<0.05 respectively) in the small intestine of aged rutin-fed mice. Meanwhile, there was no change in interferon γ gene expression between control and rutin-fed mice. IL-13 gene expression was also downregulated in the lung. To examine the mechanism of the inhibitory effect of rutin on Th2 cytokines in aged mice, intestinal nitric oxide synthase (NOS) expression was evaluated. Rutin inhibited inducible NOS (NOS2) gene expression, but not neuronal NOS and endothelial NOS. Gene analysis of cells collected from the small intestine by laser capture dissection revealed that NOS2 expression was significantly inhibited in crypt regions. Thus, rutin might be effective against a Th2-dominant profile through NOS2 inhibition in aged mice.     

Cynomolgus macaque CYP4 isoforms are functional, metabolizing arachidonic acid

Cytochrome P450 (CYP) is important for metabolism of not only xenobiotics such as drugs, but also endogenous compounds including arachidonic acids. CYP4A11, CYP4F3v2, CYP4F11, and CYP4F45 have been identified in cynomolgus macaque, an animal species widely used for investigation of drug metabolism due to its evolutionary closeness to human. However, their metabolic functions have not been investigated. In this study, proteins were heterologously expressed in Escherichia coli and characterized by metabolic assays using arachidonic acids as substrates that are metabolized by CYP4 isoforms in human. The results showed that all four CYPs metabolized arachidonic acids. Therefore, cynomolgus macaque CYP4A11, CYP4F3v2, CYP4F11, and CYP4F45 are functional enzymes.     

Absorption and emission of CO2 by ponded water of a paddy field

In the daytime, the CO₂ concentration in the air close to the water surface of a ponded paddy field was lowest and it increased with the distance above the water surface, while an inverse relation was observed in the nighttime. On the other hand, the pH of the ponded water changed significantly throughout a day and was expected to affect atmospheric CO₂ in the vicinity of the water surface, because the solubility of CO₂ in water depends on the pH. In this study, we investigated the relationship between the changes in the pH of the ponded water and the response of the CO₂ concentration in the air above the water. The pH of the ponded water of the paddy field increased in the daytime and decreased in the nighttime, so that the water was alkaline in the daytime and weakly acidic in the nighttime. We found that the daily changes in the atmospheric CO₂ concentration gradient almost corresponded to the daily changes in pH. The increase of the pH of the ponded water in the daytime was due to the absorption of dissolved CO₂ by photosynthetic bacteria and micro-algae within the ponded water. Furthermore, we compared the pH with RpH, defined as the pH at which the CO₂ concentration of the water is in equilibrium with that of the air, to determine whether CO₂ was absorbed by or emitted from the ponded water. In the daytime, the pH value of the ponded water was higher than that of the RpH, and the water could therefore absorb CO₂ , whereas during the nighttime, since the pH value of the ponded water was lower than that of the RpH, the water was expected to emit CO₂. These results show that the ponded water absorbed CO₂ from the air above the water surface in the daytime and emitted CO₂ in the nighttime.     

Rice phenolics efflux transporter 2 (PEZ2) plays an important role in solubilizing apoplasmic iron

Iron (Fe) is an essential micronutrient for plants; however, despite its abundance in soils it is not readily available in neutral to alkaline soils. Plants secrete phenolics, such as protocatechuic acid (PCA) and caffeic acid (CA), to absorb and utilize precipitated apoplasmic Fe from root surfaces. However, the synthesis and secretion of phenolics have not been well characterized in plants. We have identified and characterized a rice (Oryza sativa L.) mutant with reduced amounts of PCA and CA in xylem sap and root exudates; hence we named it phenolics efflux zero 2 (pez2). PEZ2 localized to the plasma membrane in onion (Allium cepa L.) epidermal cells and transported PCA when expressed in Xenopus laevis oocytes. PEZ2 expression was observed in whole root near root tips. Similarly, strong expression was observed in leaves. In line with reduced amounts of PCA and CA in xylem sap, the xylem Fe concentration was also low in pez2 plants. These results suggest that PEZ2 is involved in solubilization of apoplasmic Fe in rice.     

Influence of soil-drying under field conditions on exchangeable manganese,cobalt, and copper contents

Abstract

The changes in the exchangeable Mn (Ex-Mn), exchangeable Co (Ex-Co), and exchangeable Cu (Ex-Cu) contents by air-drying of soils were reported in our previous paper. The drying effect, however, was not analyzed in the field. The objective of this study was to investigate the influence of soil-drying under field conditions on Ex-Mn, Ex-Co, and Ex-Cu levels. Two paddy soils (Tsukuba and Yawara) were collected at four different soil depths (0?1, 1?5, 5?10, and 10?20 cm) under three different soil moisture conditions i.e., after harvest, after successive sunny days and after rainy days. After sunny days, Ex-Mn and Ex-Co contents in the surface soil layer markedly increased and drastically decreased after rainfall, while they remained almost unchanged in the lower soil layers. Ex-Cu contents slightly increased in the surface soil layer after sunny days. A similar tendency was observed in soil column experiments. The contents of Ex-Mn, Ex-Co, and Ex-Cu increased 12, 12, and 2 times in the upper layer of soil columns that had been sampled from the Yawara field and stored in a dark room for 8 d. The increase of the Ex-Mn and Ex-Co contents in the upper layer could not be explained by the capillary rise of soil water from the lower layers of the columns. In a soil-drying experiment under laboratory conditions, Ex-Mn and Ex-Co contents began to increase when the water content of the Yawara soil was lower than 0.09?0.13 kg kg^?1. The water content of the surface soil of the Yawara field after sunny days was 0.08 kg kg^?1. Ex-Cu content did not change apprecially with the water content. It is concluded that the increase of the Ex-Mn and Ex-Co levels in field surface soils after sunny days can be ascribed to soil-drying. Some of the excess Ex-Mn and Ex-Co is expected to leach down into lower layers due to rain, which may contribute to plant uptake of Mn and Co.     

Detection of Tobamoviruses from Soils by Non-precoated Indirect ELISA

A method for detecting tobamoviruses from field soils was developed using non-precoated indirect enzyme-linked immunosorbent assay (Id-ELISA). Absorbance values in Id-ELISA were relatively low after directly applying Pepper mild mottle virus (PMMoV)-infested soil extract. However, heat treating the soil extract before application greatly enhanced the absorbance values. The heat treatment was essential for the Id-ELISA detection of tobamoviruses from infested soil, although the efficiency of virus recovery varied depending on the properties of soil. The number of local lesions in the infectivity assay was consistent with the absorbance values in Id-ELISA. Moreover, the absorbance values in Id-ELISA were correlated with the incidence of soil transmission of PMMoV. Thus, Id-ELISA combined with heat treatment is a practical technique for the diagnosis of infestation with Tobamovirus in field soils, Gray Lowland soil and Sand-dune Regosol. Received 4 October 1999/ Accepted in revised form 9 December 1999     

Comparison of rDNA-ITS Sequences between Potato and Tobacco Strains in <Emphasis Type="Italic">Rhizoctonia solani</Emphasis> AG-3

Genetic diversity among 51 isolates of Rhizoctonia solani AG-3, representing potato and tobacco populations, was inferred from the sequences of the internal transcribed spacer (ITS) and 5.8S ribosomal RNA (rRNA) gene. The 5.8S rDNA sequence was completely conserved not only in AG-3, but across all the AG isolates examined, whereas the rDNA-ITS sequence was found to be variable among the isolates. The nucleotide sequence similarity in the ITS 1 region was high (96-100%) for isolates within each of the two populations, but was 91-92% for isolates from different populations. The AG-3 isolates had 56 to 91% sequence similarities in the ITS 1 region with R. solani isolates of the other AGs. Phylogenetic analysis based on the ITS-5.8S rDNA sequence data indicated that the different populations in AG-3 are distantly related to each other. Genetic divergence between the two populations was also supported by the results of DNA-DNA hybridization studies. This study suggests that AG-3 consists of two genetically isolated groups corresponding to separate subgroups: AG-3 PT (potato type) and AG-3 TB (tobacco type). Specific primer sets for the detection of the two AG-3 subgroups were developed from the aligned rDNA-ITS sequences. Received 22 April 1999/ Accepted in revised form 2 July 1999