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Supravalvular aortic stenosis as a late complication of transposition of the great arteries is very rare, and only a few cases have been reported. We describe the case of a 14-year-old girl who developed supravalvular aortic stenosis as a late complication of the arterial switch operation for transposition of the great arteries. The narrowed ascending aorta was replaced with a graft. The right pulmonary artery was transected to approach the ascending aorta which adhered severely to the main pulmonary trunk, and we obtained a good operative field. [ABSTRACT FROM AUTHOR]

Sacotoxin IA, an anti-microbial peptide of insects, inhibited the growth of harmful human intestinal bacteria such as Escherichia coli, Clostridiumramosum and Clostridium paraputrificum, in vitro. Interestingly, E. coli O157 which is well-known to cause food poisoning, was most sensitive to the peptide among the bacteria tested. The peptide, however, did not affect the growth of Bifidobacterium adolescentis, Bifidobacterium longum, Lactobacillus acidophilus, Enterococcus faecalis, Bacteroides thetaiotamicron, Bacteroides uniformis and Bacteroides ovatuss which are abundant in the intestines of healthy people. The evidence suggests that sarcotoxin IA could change the composition of the intestinal flora to improve human health. [ABSTRACT FROM AUTHOR]

L. (cv. Petit Havana SR1) were grown under ultraviolet-B (UV-B, 290–320 nm) irradiation, and soluble proteins were extracted from the leaves. Two-dimensional electrophoresis revealed that a minimum of 12 polypeptides were induced by UV-B. Polypeptides which were so abundant as to be detectable by Coomassie brilliant blue staining were then subjected to N-terminal amino acid sequence analyses. Two of the polypeptides were identified as a 23 kDa protein of PS II and 6 as a pathogenesis-related protein 5 (PR-5). Immunoblotting demonstrated that other PR proteins, PR-1 and PR-3 were also induced by UV-B. Salicylic acid (SA), which is an important component of signal transduction that leads to the expression of PR proteins and exhibition of acquired resistance to pathogens, increased in response to exposure to UV-B. In addition, the activity of phenylalanine ammonialyase, which catalyzes the synthesis from phenylalanine of trans-cinnamic acid, the endogenous precursor of SA, was transiently increased by UV-B irradiation. These results suggest that UV-B activates the signal transduction pathway, which is a common step in pathogen infection. [ABSTRACT FROM AUTHOR]

A cDNA clone encoding a cysteine protease was isolated from a tobacco cDNA library, utilizing as a probe a PCR fragment obtained from degenerated primers based on the conserved sequences of plant cysteine protease genes. A putative protein encoded by the clone NTCP-23 had an amino acid sequence with significant similarities to those of plant senescence-associated cysteine proteases and mammalian cathepsin H. Northern blot analysis showed that NTCP-23 mRNA is expressed in all organs and the mRNA and protein expression is enhanced during natural senescence. We propose that NTCP-23 is responsible for amino acid remobilization especially in senescencing leaves. Furthermore, it was found that the mRNA expression follows a circadian rhythm and is reduced by continuous darkness, wounding and hypersensitive reaction (HR). NTCP-23 is the first cysteine protease whose mRNA expression has been shown to be temporarily reduced by wounding. [ABSTRACT FROM AUTHOR]

In tobacco plants, wounding induces production of a set of defense-related proteins such as basic pathogenesisrelated (PR) proteins and proteinase inhibitors (PIs) via the jasmonate/ethylene pathway. Although class III plant peroxidase (POX) is also wound-inducible, the regulatory mechanism for its wound-induced expression is not fully understood. Here, we describe that a tobacco POX gene (tpoxN1), which is constitutively expressed in roots, is induced locally 30 min after wounding and then systemically in tobacco plants. Infection of necrotizing virus also induced tpoxN1 gene. The wound-induced expression was not enhanced by known wound-signal compounds such as methyl jasmonate (MeJA) and ethephon in contrast to other wound-inducible genes such as basic PR-1 and PI-II genes. And treatment with MeJA and coronatine, biological analogs of jasmonate, rather suppressed the tpoxN1 expression. Salicylic acid, an antagonist of jasmonatebased wound signaling, did not suppress the wound-induced expression of tpoxNl. Only spermine, which is reported as an endogenous inducer for acidic PR genes in tobacco mosaic virus-infected tobacco leaves, could induce tpoxN1 gene expression. These results suggest that woundinduced expression of the tpoxN1 gene is regulated differently from that of the basic PR and PI-II genes. [ABSTRACT FROM AUTHOR]

Background: With the increasing number of patients with Alzheimer's disease, construction of inspection system for blood biomarkers to replace or complement amyloid PET and CSF biomarkers has been required. Since our first report in 2014 (Kaneko et al., Proc Jpn Acad Ser B Phys Biol Sci. 2014), we have been focusing on three amyloid peptides, Aβ1‐40, Aβ1‐42, and APP669‐711 in human plasma, and found that the combination of peptide ratios APP669‐711/Aβ1‐42 and Aβ1‐40/Aβ1‐42 became a biomarker for amyloid peptide accumulation in human brain (Nakamura et al., Nature 2018). Here we describe the analytical validation of plasma amyloid β measuring system by immunoprecipitation‐matrix‐assisted laser desorption ionization time‐of‐flight mass spectrometry (IP‐MALDI‐MS). Method: Commercially available human plasma and artificial plasma were immunoprecipitated with anti‐Aβ antibody (clone 6E10) covalently bound to the magnetic beads, then measured by MALDI‐TOF‐MS. The peak intensity of the Aβ‐related peptides was normalized by the internal standard peak, and biomarker values were calculated from those intensities. A method validation including experiments of calibration range, sensitivity, specificity, reproducibility, interfering substances, dilution linearity, recovery, and stability was performed. Result: The system showed good linearity over the range from 15 to 150 pM for Aβ1‐40, and 3 to 15 pM for Aβ1‐42 and APP669‐711. The trueness was within 100 ± 25% at the lower limit of quantification and within 100 ± 20% at all other concentrations. The precision of the intra‐ and inter‐assay was 1% to 13% as a CV value. Recovery from spiked human plasma ranged from 83% to 101% at low, middle, and high concentrations. Stability testing has shown that the antibody beads are stable for up to 6 months in the refrigerator so far. The addition of the interfering substances, such as anticoagulants, Alzheimer's disease drugs, etc., had only a slight effect on the biomarker values showing 10% or less difference relative to the control. Further results will be reported in the presentation. Conclusion: The results demonstrate that the plasma amyloid β measuring system by IP‐MALDI‐MS is an accurate, reliable, and robust assay that would be most suitable for screening subjects in clinical trials. [ABSTRACT FROM AUTHOR]

Despite improvements in bioprosthetic valve function, increased human life-expectancy has led to a growing number of bioprosthetic valve deterioration cases requiring reoperation. We report 2 cases of primary tissue failure of a bioprosthetic valve, which were treated by mitral valve replacement using the valve-on-valve method. The reasons for the reoperations were a severely calcified valve annulus, and severe adhesion of a previous bioprosthetic valve. We removed only the leaflets of the bioprosthetic valve and sutured a Carbomedics OptiForm valve onto the sewing cuff of the previous bioprosthesis. No complications, including major cardiac events, were noted during the follow-up. [ABSTRACT FROM PUBLISHER]

Ethylene has been thought to be an important factor for the induction of defense responses against pathogen attack, but its function in these responses has not been well defined. Here we show the direct involvement of ethylene in the formation of necrotic lesions and in basic pathogenesis-related (PR) protein gene induction in tobacco. Increase in ethylene production as well as accumulation of 1-aminocyclopropane-1-carboxylic acid oxidase (ACO) gene transcript obviously preceded the lesion appearance in tobacco mosaic virus (TMV)-infected leaves in an N gene-dependent synchronous lesion formation system. Inhibitors of ethylene biosynthesis or action significantly suppressed both lesion formation and basic PR gene expression. Induction of these genes was enhanced in 1-aminocyclopropane-1-carboxylic acid (ACC)-treated leaves especially in ACO-overexpressing transgenic tobacco plants. Further, we found that ethylene production during hypersensitive reaction is restricted at the level of ACO activity. [ABSTRACT FROM AUTHOR]

Treatment of wounded tobacco leaves with a proteasome-specific inhibitor lactacystin resulted in; (1) enhanced expression of proteasome subunit genes, (2) suppressed expression of proteasome activity, (3) induced accumulation of Ub-conjugated proteins, and (4) suppressed expression of wound-inducible genes, indicating that the Ub-proteasome system is involved in wound-signaling in plants. [ABSTRACT FROM AUTHOR]

Sarcotoxin IA is a bactericidal peptide of 39 amino acids found in the common flesh fly, Sarcophaga peregrina. Many agronomically important bacteria in Japan are killed by this peptide at sab-micro molar levels, and the growth of tobacco and rice suspension cultured cells is not inhibited with less than 25 µM. Transgenic tobacco plants which overexpress the peptide, i.e. over 250 pmol per gram of fresh leaf, under the control of a high expression constitutive promoter showed enhanced resistance to the pathogens for wild fire disease (Pseudomonas syringae pv. tabaci) and bacterial soft rot disease (Erwinia carotovora subsp. carotovora). [ABSTRACT FROM PUBLISHER]

Wounding induces expression of genes encoding defense-related proteins involved in wound healing. An intensive survey has been carried out to clarify the initial signal transduction pathways that mediate this stress to expression of genes. In this context, signal molecules that intermediate in the wound signal to cellular response have been actively searched for. Jasmonic acid (JA) has been considered to be a key signal molecule in this pathway. Systemin, ABA, ethylene, and electrical current have been suggested to function by transmitting the wound signal to JA. A mitogen-activated protein kinase has been shown to respond rapidly to wounding, and proposed to function as one of the key enzymes involved in JA biosynthesis. Transgenic plants overexpressing a gene encoding a Rab-type, small GTP-binding protein contained 6-fold higher levels of cytokinins than wild-type plants, and responded to wounding by rapidly producing JA and, uncommonly, accumulating salicylic acid (SA), a pathogenic signal. These phenomena observed in the transgenic plants were reproduced when wild-type plants were wounded in the presence of the synthetic cytokinin, benzylaminopurine, suggesting that cytokinins are indispensable in the control of endogenous levels of JA and SA. [ABSTRACT FROM AUTHOR]

Salicylic acid (SA) and jasmonic acid (JA) are essential compounds in the pathogen- and wound-signaling pathways accompanying induced expression of acidic and basic pathogenesis-related (PR) protein genes, respectively. However, on the effect of exogenously supplied SA and JA in induction of PR gene expression, conflicting results have been obtained using various plant materials at different developmental stages. There is no clear evidence on these effects in the presence of both signals at the same time. We analyzed the effect of SA on wound- and JA-induced basic PR gene expression and that of JA on SA-induced acidic Pi? gene expression in mature tobacco leaves. Wound-induced accumulation of transcripts for all 4 basic PR genes tested was enhanced in the presence of MeJA, and inhibited in the presence of SA. On the other hand, expression of all 3 acidic PR genes tested was induced by SA and was inhibited by MeJA. Using antibodies raised against acidic PR-1 and PR-2 proteins, these effects were confirmed at the protein level. These results indicated that JA works as an inducer of basic PR genes, and also as an inhibitor for acidic PR genes, while SA does the opposite. [ABSTRACT FROM AUTHOR]

To enhance the disease resistance of plants expressing a foreign peptide, the gene for sarcotoxin IA, which is an antimicrobial peptide from an insect consisting of 39 amino acid residues, was introduced into tobacco (Nicotiana tabacum) under the control of a high expression promoter via Agrobacterium-mediated transformation. In transgenic plants, sarcotoxin IA mRNA accumulated to detectable levels, however, the amount of the peptide produced was so small that we could scarcely detect it by protein gel blot analysis, probably because of the instability of short peptides in plant cells. To improve the expression efficiency, genes for four types of amino-terminal and carboxyl-terminal translational fusions of sarcotoxin IA together with the GUS gene were introduced into tobacco. In all four types of transgenic tobacco plants, high level transcripts similar to that in the direct expression sarcotoxin IA construct were found. Protein gel blot analysis with both anti-sarcotoxin IA and GUS antibodies showed production of high levels of fusion protein in all transgenic plants. Among them, three types had abnormal membranes and phenotypes, although no such abnormalities were found in transgenic plants in which only sarcotoxin IA was expressed in a secretable form. All together, these results indicated that, for stable and effective expression of a foreign short peptide in transgenic plants, expression as a fusion protein is useful and that secretion of sarcotoxin IA outside of cells is necessary for generation of useful antimicrobial transgenic plants. [ABSTRACT FROM AUTHOR]

Plants respond differentially to wounding and pathogens using distinct signaling pathways, so that wound signals are transmitted to jasmonic acid (JA) which induces basic pathogenesis-related (PR) proteins, whereas pathogenic signals cause, in addition to JA, accumulation of salicylic acid (SA) which stimulates production of acidic PR proteins. Transgenic tobacco plants expressing a gene for a small GTP binding protein respond abnormally to mechanical wounding to produce SA and consequently acidic PR proteins, suggesting that wound signals cross with pathogen signaling pathways [Sano et al. (1994) Proc. Natl. Acad. Sci. USA 91: 10556]. This unusual signal crossing is associated with a highly sensitive wound-response of transgenic plants which, upon wounding, produce JA at least eighteen hours earlier than wild-type plants. When wildtype plants are wounded in the presence of the synthetic cytokinin, benzylaminopurine, production of JA begins six hours earlier than in untreated samples, and also SA begins to accumulate. The cytokinin antagonist, 2-chloro-4-cyclohexylamino-6-ethylamino-s-triazine, erases these effects. Because transgenic plants constitutively produce four-to six-fold higher amounts of endogenous cytokinins than wild-type plants, it is concluded that cytokinins are indispensable for control of endogenous levels of SA and JA. [ABSTRACT FROM AUTHOR]

A major target protein of antineutrophil cytoplasmic antibody with a perinuclear staining pattern (P-ANCA) has been identified as myeloperoxidase (MPO). Recombinant deletion mutants of MPO, eight fragments of the heavy-chain subunit, and two fragments of the light chain subunit were expressed in E. coli using a pQE expression vector. The recombinant hexamer histidine-tagged fragments were partially purified as the denatured proteins on a Ni2+-charged nitrirotriacetic acid column. The recombinant fragments were reacted with a rabbit polyclonal antibody to human MPO in Western blotting. In addition, the reactivities of the proteins with MPO-ANCA-positive sera of four patients with renal diseases were examined by Western blotting. The profile of the reactivity showed that different sera recognized different sets of fragments of the heavy chain, whereas no serum reacted with the fragments of the light chain. These results indicate that the sera of patients with MPO-ANCA-positive diseases showed varied reactivities with the different fragments. Furthermore, an ELISA system using a set of the fragments completely purified by Sephacryl S-200HR column chromatography was established. The panel set is useful for subclassification of MPO-ANCA-related diseases. [ABSTRACT FROM AUTHOR]

Background: The measurement of plasma amyloid β recently attracts much attention as a biomarker of Alzheimer's disease. We developed a highly sensitive method for simultaneous detection of a plurality of amyloid β peptides in human plasma by using immunoprecipitation combined with matrix‐assisted laser desorption ionization time‐of‐flight mass spectrometry (IP‐MALDI‐MS). In this study, we examined the effects of interfering substances in human plasma to reinforce the performance and practicability of this approach. Method: Anticoagulants, Alzheimer's disease drugs, peptides having the epitope with high homology to 6E10 anti‐amyloid β antibody, and a drug having a similar mass to the amyloid β peptide were added to human plasma. The doped plasmas were then analyzed by IP‐MALDI‐MS. Plasma with PBS added was used as a control. Result: Two biomarker values (Aβ1‐40/Aβ1‐42 and APP669‐711/Aβ1‐42) in the interfering substances doped plasma were compared to those of control plasma. The results showed that the addition of any of those drugs had only a slight effect on the measured values, and the difference between the plasma biomarkers to which the compound was added was ± 10% or less relative to the control. Conclusion: The analysis of plasma amyloid β biomarkers by IP‐MALDI‐MS is a unique method for the simultaneous detection of multiple amyloid β peptides requiring only one kind of antibody. Addition of twelve kinds of contaminants to the human plasma have limited effects on the analytical performance of the system. [ABSTRACT FROM AUTHOR]