Composting of rice straw with oilseed rape cake and poultry manure and its effects on faba bean (Vicia faba L.) growth and soil properties

Composting of rice straw with poultry manure and oilseed rape cake and its effects on growth and yield of faba bean and soil properties was studied in pot experiments at Gifu University, Japan in 2001/2002. The composts reached maturity in 90 days, were rich in organic matter and mineral nutrients, had a high level of stability, and no phytotoxicity. The addition of compost (20-200 g pot(-1)) improved selected soil chemical (increased total N, total C and CEC), physical (decreased particle density) and biological (increased soil respiration rate) properties. Application of composts at a rate of 20 g pot(-1) significantly increased growth, yield, yield components and total crude protein of faba bean plants. The benefit of this compost without chemical fertilizer demonstrated the validity and possibility of sustainable agronomic performance of faba bean using locally available recycled organic materials.     

Fish gonadotropin and thyrotropin receptors: the evolution of glycoprotein hormone receptors in vertebrates

We have cloned and characterized, for the first time in fish, two different gonadotropin receptors (GTHR) and a single thyrotropin receptor (TSHR) from amago salmon (Oncorhynchus rhodurus) and Nile tilapia (Oreochromis niloticus). Phylogenetic analyses and intron/exon structure suggest that the two GTHRs in fish are comparable to tetrapod follicle stimulating hormone and luteinizing hormone receptors. Temporal and spatial expression patterns, examined by Northern blot analysis and in situ hybridization, paralleled those seen in mammals and birds. Consequently, genetic and functional divergence of two GTHRs and TSHR probably occurred before the teleost and tetrapod split.     

Generation of polyclonal antibody against mu-conotoxin GIIIA using an immunogen of [Cys(5)]mu-conotoxin GIIIA site-specifically conjugated with bovine serum albumin.

mu-Conotoxin GIIIA, one of the strong peptide toxins in the cone shell, preferentially blocks the skeletal muscle-type sodium channels in vertebrates. The toxicity of mu-conotoxin GIIIA is nearly equal to that of tetrodotoxin. The generation of an antibody for the native toxins is analytically useful, but practically difficult due to its high toxicity to animals. In this study, we generated the polyclonal antibody for mu-conotoxin GIIIA using a specific conjugation method in which the immunogen was detoxified while retaining the active-site structure for the sodium channels. ELISA analysis showed that the generated antibody recognized the native toxin folded with three disulfide bridges, but not the linear one. Furthermore, the physiologically active mutants of GIIIA were recognized while the inactive mutants were not, suggesting that the newly generated antibody can selectively recognize the physiologically active toxins. These methods for generating an antibody against peptide toxins will be applicable to other peptide toxins.     

Difference in Postural Control during Quiet Standing between Young Children and Adults: Assessment with Center of Mass Acceleration

The development of upright postural control has often been investigated using time series of center of foot pressure (COP), which is proportional to the ankle joint torque (i.e., the motor output of a single joint). However, the center of body mass acceleration (COM_acc), which can reflect joint motions throughout the body as well as multi-joint coordination, is useful for the assessment of the postural control strategy at the whole-body level. The purpose of the present study was to investigate children’s postural control during quiet standing by using the COM_acc. Ten healthy children and 15 healthy young adults were instructed to stand upright quietly on a force platform with their eyes open or closed. The COM_acc as well as the COP in the anterior–posterior direction was obtained from ground reaction force measurement. We found that both the COM_acc and COP could clearly distinguish the difference between age groups and visual conditions. We also found that the sway frequency of COM_acc in children was higher than that in adults, for which differences in biomechanical and/or neural factors between age groups may be responsible. Our results imply that the COM_acc can be an alternative force platform measure for assessing developmental changes in upright postural control.     

DNA Barcoding of Japanese Click Beetles (Coleoptera,Elateridae)

Click beetles (Coleoptera: Elateridae) represent one of the largest groups of beetle insects. Some click beetles in larval form, known as wireworms, are destructive agricultural pests. Morphological identification of click beetles is generally difficult and requires taxonomic expertise. This study reports on the DNA barcoding of Japanese click beetles to enable their rapid and accurate identification. We collected and assembled 762 cytochrome oxidase subunit I barcode sequences from 275 species, which cover approximately 75% of the common species found on the Japanese main island, Honshu. This barcode library also contains 20 out of the 21 potential pest species recorded in Japan. Our analysis shows that most morphologically identified species form distinct phylogenetic clusters separated from each other by large molecular distances. This supports the general usefulness of the DNA barcoding approach for quick and reliable identification of Japanese elaterid species for environmental impact assessment, agricultural pest control, and biodiversity analysis. On the other hand, the taxonomic boundary in dozens of species did not agree with the boundary of barcode index numbers (a criterion for sequence-based species delimitation). These findings urge taxonomic reinvestigation of these mismatched taxa.     

Gene transfer using micellar nanovectors inhibits choroidal neovascularization in vivo

Purpose

Age-related macular degeneration caused by choroidal neovascularization (CNV) remains difficult to be treated despite the recent advent of several treatment options. In this study, we investigated the in vivo angiogenic control by intravenous injection of polyion complex (PIC) micelle encapsulating plasmid DNA (pDNA) using a mice CNV model.

Methods

The transfection efficiency of the PIC micelle was investigated using the laser-induced CNV in eight-week-old male C57 BJ/6 mice. Firstly, each mouse received intravenous injection of micelle encapsulating pDNA of Yellow Fluorescent Protein (pYFP) on days 1,3 and 5. The expression of YFP was analyzed using fluorescein microscopy and western blotting analysis. In the next experiments, each mouse received intravenous injection of micelle encapsulating pDNA of soluble Fms-like tyrosine kinase-1 (psFlt-1) 1,3 and 5 days after the induction of CNV and the CNV lesion was analyzed by choroidal flatmounts on day 7.

Results

Fluorescein microscopy and western blotting analysis revealed that the expression of YFP was confirmed in the CNV area after injection of the PIC micelle, but the expression was not detected neither in mice that received naked pDNA nor those without CNV. Furthermore, the CNV area in the mice that received intravenous injection of the psFlt-1-encapsulated PIC micelle was significantly reduced by 65% compared to that in control mice (p<0.01).

Conclusions

Transfection of sFlt-1 with the PIC micelle by intravenous injection to mice CNV models showed significant inhibition of CNV. The current results revealed the significant potential of nonviral gene therapy for regulation of CNV using the PIC micelle encapsulating pDNA.     

Tight junction regulates epidermal calcium ion gradient and differentiation

It is well known that calcium ions (Ca²⁺) induce keratinocyte differentiation. Ca²⁺ distributes to form a vertical gradient that peaks at the stratum granulosum. It is thought that the stratum corneum (SC) forms the Ca²⁺ gradient since it is considered the only permeability barrier in the skin. However, the epidermal tight junction (TJ) in the granulosum has recently been suggested to restrict molecular movement to assist the SC as a secondary barrier. The objective of this study was to clarify the contribution of the TJ to Ca²⁺ gradient and epidermal differentiation in reconstructed human epidermis. When the epidermal TJ barrier was disrupted by sodium caprate treatment, Ca²⁺ flux increased and the gradient changed in ion-capture cytochemistry images. Alterations of ultrastructures and proliferation/differentiation markers revealed that both hyperproliferation and precocious differentiation occurred regionally in the epidermis. These results suggest that the TJ plays a crucial role in maintaining epidermal homeostasis by controlling the Ca²⁺ gradient.