Catheter-based antegrade intracoronary viral gene delivery with coronary venous blockade

The purpose of this study is to evaluate the feasibility of percutaneous antegrade myocardial gene transfer (PAMGT). A consistent and safe technique for in vivo gene transfer is required for clinical application of myocardial gene therapy. PAMGT with concomitant coronary venous blockade was performed in 12 swine. The myocardium was preconditioned with 1 min of occlusion of the left anterior descending and left circumflex arteries. The anterior interventricular vein was occluded during left anterior descending artery delivery, and the great cardiac vein at the entrance of the middle cardiac vein was occluded during left circumflex artery delivery. With arterial and venous balloons inflated (3 min) and after adenosine (25 mug) injection, PAMGT was performed by antegrade injection of an adenoviral solution (1 ml of 10(11) plaque-forming units in each coronary artery) carrying beta-galactosidase or saline through the center lumen of the angioplasty balloon. In one set of animals, PAMGT was performed with selective coronary vein blockade (n = 9); in another set of animals, PAMGT was performed without coronary vein blockade (n = 5). At 1 wk after gene delivery, the animals were killed. Quantitative beta-galactosidase analysis was performed in the left and right ventricular walls. PAMGT was successfully performed in all animals with and without concomitant occlusion of the coronary veins. Quantitative beta-galactosidase analysis showed that PAMGT with coronary blockade was superior to PAMGT without coronary blockade. beta-Galactosidase activity increased significantly in the beta-galactosidase group compared with the saline group: 1.34 +/- 0.18 vs. 0.81 +/- 0.1 ng (P 相似文献   

Two different types of hepcidins from the Japanese flounder Paralichthys olivaceus

The cysteine-rich peptide hepcidin is known to be an antimicrobial peptide and iron transport regulator that has been found in both fish and mammals. Recently, we found two different types (designated Hep-JF1 and Hep-JF2) of hepcidin cDNA in the Japanese flounder, Paralichthys olivaceus, by expressed sequence tag analysis. The identity of amino acid sequences between Hep-JF1 and Hep-JF2 was 51%. The Hep-JF1 and Hep-JF2 genes both consist of three exons and two introns, and both exist as single copies in the genome. The predicted mature regions of Hep-JF1 and Hep-JF2 have six and eight Cys residues, respectively. The first Cys residue of Hep-JF1 was deleted and the second was replaced with Gly. The number and positions of Cys residues in Hep-JF2 are the same as they are in human Hep. Hep-JF1 is specifically expressed in liver while the expression of Hep-JF2 was detected from gill, liver, heart, kidney, peripheral blood leucocytes, spleen and stomach. Gene expression of Hep-JF1 in liver decreased during experimental iron (iron-dextran) overload. Expression of Hep-JF1 in liver was decreased by injecting fish with iron-dextran and increased by injecting lipopolysaccharide. Iron overload did not significantly affect expression of Hep-JF2 in liver but it did increase expression of Hep-JF2 in kidney. Lipopolysaccharide injection increased expression of Hep-JF2 in both liver and kidney. In liver, some cells expressed both Hep-JF1 and Hep-JF2 while some other cells expressed just one of them. Synthesized Hep-JF2 peptide showed antimicrobial activity, while synthesized Hep-JF1 peptide did not against several bacteria including fish-pathogenic bacteria used in this study.     

Functional dissection of two Arabidopsis PsbO proteins: PsbO1 and PsbO2

PsbO protein is an extrinsic subunit of photosystem II (PSII) and has been proposed to play a central role in stabilization of the catalytic manganese cluster. Arabidopsis thaliana has two psbO genes that express two PsbO proteins; PsbO1 and PsbO2. We reported previously that a mutant plant that lacked PsbO1 (psbo1) showed considerable growth retardation despite the presence of PsbO2 [Murakami, R., Ifuku, K., Takabayashi, A., Shikanai, T., Endo, T., and Sato, F. (2002) FEBS Lett523, 138-142]. In the present study, we characterized the functional differences between PsbO1 and PsbO2. We found that PsbO1 is the major isoform in the wild-type, and the amount of PsbO2 in psbo1 was significantly less than the total amount of PsbO in the wild-type. The amount of PsbO as well as the efficiency of PSII in psbo1 increased as the plants grew; howeVER, it neVER reached the total PsbO level observed in the wild-type, suggesting that the poor activity of PSII in psbo1 was caused by a shortage of PsbO. In addition, an in vitro reconstitution experiment using recombinant PsbOs and urea-washed PSII particles showed that oxygen evolution was better recoVERed by PsbO1 than by PsbO2. Further analysis using chimeric and mutated PsbOs suggested that the amino acid changes Val186-->Ser, Leu246-->Ile, and Val204-->Ile could explain the functional difference between the two PsbOs. Therefore we concluded that both the lower expression level and the inferior functionality of PsbO2 are responsible for the phenotype observed in psbo1.     

Land-use diversity index: a new means of detecting diversity at landscape level

Preservation of ecological diversity is a guiding principle of landscape management and planning. Although diversity is often quantified with measurable indices, common approaches used to measure diversity, for example the Shannon–Weaver index, are not adequate for many landscape studies, because they are affected by scale and sampling efforts. A robust index for measurement of landscape diversity should be able to quantify distinctive components of a landscape mosaic. This paper provides a new diversity index for landscape studies that improves the measurement of diversity at landscape level. The key features of the method are inclusion of an implication of physical moment and quantification of diversity by placing greater weight on the structure and composition of a patch. We have named the new index the land-use diversity index, or LUDI, and have concluded that the LUDI is the preferred measure of diversity at landscape level.     

The mir‐7 and bag of marbles genes regulate Hedgehog pathway signaling in blood cell progenitors in Drosophila larval lymph glands

Hedgehog (Hh) pathway signaling is crucial for the maintenance of blood cell progenitors in the lymph gland hematopoietic organ present in Drosophila third instar larvae. Previous studies from our lab have likewise shown the importance of the mir‐7 and bag of marbles (bam) genes in maintaining the progenitor state. Thus, we sought to investigate a possible interaction between the Hh pathway and mir‐7/bam in the prohemocyte population within this hematopoietic tissue. Gain of function mir‐7 was able to rescue a blood cell progenitor depletion phenotype caused by Patched (Ptc) inhibition of Hh pathway signaling in these cells. Similarly, expression of a dominant/negative version of Ptc was able to rescue the severe reduction of prohemocytes due to bam loss of function. Furthermore, we demonstrated that Suppressor of fused [Su(fu)], another known inhibitor of Hh signaling, likely serves as a translational repression target of the mir‐7 miRNA. Our results suggest the mir‐7/bam combination regulates the Hh signaling network through repression of Su(fu) to maintain hemocyte progenitors in the larval lymph gland.     

Evaluation and Allocation of Greenhouse Gas Reductions in Industrial Symbiosis

Industrial symbiosis (IS) exchanges have been recognized to reduce greenhouse gas (GHG) emission, though methods for quantification of GHG emissions in IS exchanges are varied, and no standardized methods are available. This article proposes a practical approach to quantify total and allocated GHG emissions from IS exchanges by integrating the GHG protocol and life cycle assessment. The proposed method expands the system boundaries to include all IS companies, and the functional flow is set to be the sum of the main products. The total impact of a company is allocated to the main product. Three by‐product impact allocation methods of cutoff, avoidance, and 50/50 are proposed, and the total and distributed impacts of the IS systems in an industrial park are theoretically derived. The proposed method was tested to quantify GHG reduction in a real IS exchange developed between Korea Zinc (a zinc smelter) and Hankook Paper (a paper mill company) in the Ulsan Eco‐Industrial Park initiative. The total reduction of GHG emissions in this IS exchange, 60,522 tonnes of carbon dioxide per year, was the same in the GHG protocol, whereas GHG distribution between two companies depended on the allocation method. Given that the reduction of GHG emissions from IS exchanges is the product of the collaboration of giving companies and receiving companies, the 50/50 allocation method is best from an equivalent‐responsibility and benefit‐sharing perspective. However, this study suggests a more practical implementation approach based on a flexible and negotiable method of allocating the total GHG reduction between stakeholders.     

Temporal variability of algicidal and growth-inhibiting bacteria at an eelgrass bed in the Ariake Sea,Japan

Abstract

Temporal changes of algicidal and growth-inhibiting bacteria on the fish-killing raphidophyte flagellate, Chattonella antiqua, at an eelgrass (Zostera marina) bed in southern Ariake Sea, Japan in 2011 was investigated. The maximum value (5.1?×?10⁷ CFU g^?1 wet leaf) of algicidal bacteria (AB) was detected from a biofilm formed on Z. marina on August 1 when AB in the adjacent seawater had also peaked (1.2?×?10⁴ CFU mL^?1). Two causative bacteria isolated from the biofilm and seawater on August 1 were both identified to be of the genus Alteromonas (γ-proteobacteria). AB and growth-inhibiting bacteria (GIB) were present from the beginning of sampling (May 20) to August 26, fluctuating between 8.6?×?10² and 1.2?×?10⁴, 1.2?×?10³ and 9.3?×?10³ CFU mL^?1, respectively. The highest phytoplankton density observed was 6423 cells mL^?1 on September 29 and was comprised of centric diatoms such as Chaetoceros, Skeletonema, and Thalassiosira and coincided with the absence of AB and GIB where the decline of Z. marina was also observed. These findings provide a new ecological insight on AB and GIB associated with Z. marina beds, indicating eelgrass beds have the important role as the nursery of those bacteria that can be utilized as mitigation measures of harmful algal blooms (HABs) in the future.     

Synthesis and evaluation of N-(4-benzylphenyl)piperazines as VGF inducers

A series of compounds was discovered that induce the production of VGF mRNA in SH-SY5Y cells and exhibit cytoprotection under tunicamycin induced endoplasmic reticulum (ER) stress. The aminophenol ring and linker chain of the template SUN N8075 (1) was modified to yield compounds with higher efficacy and lower propensity for adverse effects.     

Structure-based design,synthesis, and binding mode analysis of novel and potent chymase inhibitors

Based on insight from the X-ray crystal structure of human chymase in complex with compound 1, a lactam carbonyl of the diazepane core was exchanged with O-substituted oxyimino group, leading to amidoxime derivatives. This modification resulted in highly potent chymase inhibitors, such as O-phenylamidoxime 5f. X-ray crystal structure analysis indicated that compound 5f induced movement of the Leu99 and Tyr94 side chains at the S2 site, and the increase in inhibitory activity of O-phenyl amidoxime derivatives suggested that the O-phenyl moiety interacted with the Tyr94 residue. Surface plasmon resonance experiments showed that compound 5f had slower association and dissociation kinetics and the calculated residence time of compound 5f to human chymase was extended compared to that of amide compound 1.