A plant peptide encoded by CLV3 identified by in situ MALDI-TOF MS analysis

The Arabidopsis CLAVATA3 (CLV3) gene encodes a stem cell-specific protein presumed to be a precursor of a secreted peptide hormone. Matrix-assisted laser desorption/ionization-time-of-flight mass spectrometry (MALDI-TOF MS) applied to in situ Arabidopsis tissues determined the structure of a modified 12-amino acid peptide (MCLV3), which was derived from a conserved motif in the CLV3 sequence. Synthetic MCLV3 induced shoot and root meristem consumption as cells differentiated into other organs, displaying the typical phenotype of transgenic plants overexpressing CLV3. These results suggest that the functional peptide of CLV3 is MCLV3.     

Angiotensin I Converting Enzyme Inhibitory Peptides Derived from Phycobiliproteins of Dulse Palmaria palmata

We examined the inhibitory activity of angiotensin I converting enzyme (ACE) in protein hydrolysates from dulse, Palmaria palmata. The proteins extracted from dulse were mainly composed of phycoerythrin (PE) followed by phycocyanin (PC) and allophycocyanin (APC). The dulse proteins showed slight ACE inhibitory activity, whereas the inhibitory activity was extremely enhanced by thermolysin hydrolysis. The ACE inhibitory activity of hydrolysates was hardly affected by additional pepsin, trypsin and chymotrypsin treatments. Nine ACE inhibitory peptides (YRD, AGGEY, VYRT, VDHY, IKGHY, LKNPG, LDY, LRY, FEQDWAS) were isolated from the hydrolysates by reversed-phase high-performance liquid chromatography (HPLC), and it was demonstrated that the synthetic peptide LRY (IC₅₀: 0.044 μmol) has remarkably high ACE inhibitory activity. Then, we investigated the structural properties of dulse phycobiliproteins to discuss the origin of dulse ACE inhibitory peptides. Each dulse phycobiliprotein possesses α-subunit (Mw: 17,477–17,638) and β-subunit (Mw: 17,455–18,407). The sequences of YRD, AGGEY, VYRT, VDHY, LKNPG and LDY were detected in the primary structure of PE α-subunit, and the LDY also exists in the APC α- and β-subunits. In addition, the LRY sequence was found in the β-subunits of PE, PC and APC. From these results, it was suggested that the dulse ACE inhibitory peptides were derived from phycobiliproteins, especially PE. To make sure the deduction, we carried out additional experiment by using recombinant PE. We expressed the recombinant α- and β-subunits of PE (rPEα and rPEβ, respectively), and then prepared their peptides by thermolysin hydrolysis. As a result, these peptides showed high ACE inhibitory activities (rPEα: 94.4%; rPEβ: 87.0%). Therefore, we concluded that the original proteins of dulse ACE inhibitory peptides were phycobiliproteins.     

Correlation between cytochrome P450 1A (cyp1a) mRNA expression and ambient phenanthrene and pyrene concentration in Javanese Medaka Oryzias javanicus

Fisheries Science - Cytochrome P450 1A (Cyp1a) catabolizes polycyclic organic compounds. In this study, we sequenced the cyp1a gene from two Javanese medaka Oryzias...     

Experimental study on the effect of waves on netting panels at a range of angles to the wave direction

The effect of horizontal waves on flexible netting panels placed at angles to the wave direction is studied with the aim of evaluating the testing method of pre-tensioned mooring and radial systems and flexible netting structures. The netting was hung on a frame at a specific hanging ratio for ten types of polyethylene panels. Regular waves were experimentally generated with a wave period varying from 0.8 to 2.0 s and wave height ranging from 50 to 250 mm. The force on the netting structure was recorded by a tension transducer and a digital signal recorder. The results showed that the horizontal wave force on the netting panel changed periodically and asymmetrically when it was back and left or right declinate to the wave direction; similar results were found for the surface wave elevation. The opposite results were obtained when the sloping angle declinated front to the wave direction, with two obvious crests during each period. The horizontal wave force was related to the height and width of the netting panel, wave height, wave length, twine diameter, bar length of the mesh, and sloping angle relative to the wave direction. Using dual series relations, the least square approximation, and multiple stepwise regression analysis, the formula for estimating the maximum value of the wave force on the netting was obtained.     

Development of an efficient bioreactor system for delivering foreign proteins secreted from liver into eggs with a vitellogenin signal in medaka Oryzias latipes

In this study, we developed a novel bioreactor system to deliver and accumulate foreign proteins in eggs using medaka fish Oryzias latipes with the aid of a partial sequence of vitellogenin (Vtg). In teleost fish, Vtg, the hepatically generated precursor of egg yolk proteins, is secreted into the bloodstream and then taken up into eggs. We predicted in silico a probable region (Vtg signal) of Vtg that mediates transportation of proteins from the liver into eggs. Then, we established two transgenic lines expressing the fused proteins including the Vtg signal and each reporter gene, enhanced green fluorescent protein (EGFP) or firefly luciferase (LUC)-fused EGFP, in the liver driven by a liver-specific choriogeninH (chgH) promoter. Each reporter signal was detected from the fertilized eggs spawned by the transgenic females, showing successful transportation of the proteins into the eggs with the Vtg signal. This is the first report demonstrating that the Vtg signal has capability to deliver exogenous proteins into eggs. Because Vtg is a highly conserved protein among most of oviparous organisms, our findings hold promise for establishing bioreactor systems viable in a wide range of organisms.

     

Chemical control of Haematobia irritans with 0.5% topical ivermectin solution in cattle.

A field trial was conducted to evaluate the efficacy of a topical formulation of ivermectin administered at the dose of 500 micrograms/kg against horn flies (Haematobia irritans) in cattle. Eighty-eight cattle in four herds naturally exposed to horn flies were used in the trial. Replicates were formed of two herds. Within replicates, one herd was randomly allocated to the untreated control and the other to the ivermectin treatment group. Horn fly counts were taken on the treatment day (Day 0) and on Days 7, 14, 21, 28, and 35 post-treatment. There were no horn flies on any cattle in the treatment group, whereas all the control cattle were continuously infested by horn flies on each examination day.     

Cytological relationships among Eumelilotus species analysed using cytological observations of interspecific hybrids obtained by in vitro culture

Interspecific hybrid plants between Melilotus altissima (yellow flowers) and either M. alba (white flowers) or M. taurica (white flowers) and between M. suaveolens (yellow flowers) and M. wolgica (white flowers) which were inviable because of heavy chlorosis, were grown successfully by in vitro culture and reached the flowering stage. Pollen fertility of hybrid plants between M. altissima and M. alba was about 46% whereas hybrids between M. altissima and M. taurica and between M. suaveolens and M. wolgica showed about 25% and 18% of pollen fertility, respectively. All F₁s did not set any selfed seeds. Observing the pairing of chromosomes at metaphase I in the hybrid plants, it was revealed that the genomes of M. altissima and M. alba differ by a reciprocal translocation. This result could account for the partial sterility of pollen in the F₁ hybrids. On the other hand, hybrid plants between M. altissima and M. taurica and between M. suaveolens and M. wolgica showed normal pairing of chromosomes though pollen fertility of the hybrids was very low, suggesting that low pollen fertility was controlled by sterility gene(s). These results demonstrate that M. altissima and 7 other species of the subgenus Eumelilotus share the same chromosomal constitution. The only exception, M. alba, differs from the other 8 species by a segmental interchange. This revised version was published online in July 2006 with corrections to the Cover Date.