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Open in new tab Download slide OBJECTIVES Low patency is a major concern when using separate tube grafts for intercostal artery reconstruction. Our goal was to elucidate the optimal size and length of grafts from their patency and the computational fluid dynamics (CFD). METHODS The patency, size and length of separate tube grafts were evaluated in 41 patients. Simulation of CFD was performed in a model derived from a patient with a patent 12-mm graft that was 15 mm long, with 2 simulation models with a smaller (8-mm) or longer (30-mm) graft. RESULTS A total of 49 grafts were used for intercostal artery reconstruction. There was 1 in-hospital death and 2 spinal cord injuries. The patency rate, which could be evaluated in 46 grafts, was 63% (29/46). It was 71% (24/34) in thoracoabdominal aortic replacement and 42% (5/12) in descending aortic replacement. Among 14 patients in whom all grafts were occluded, no patients developed spinal cord injury. All grafts longer than 25 mm were occluded (n = 5). Eight- and 10-mm grafts showed better patency than 12-mm grafts in thoracoabdominal aortic replacement (P  = 0.008) when grafts were shorter than 25 mm. Simulation of CFD revealed vortical flow within the 12-mm graft, which did not reach the intercostal orifice, whereas helical flow was maintained throughout the cardiac cycle within the 8-mm graft. CONCLUSIONS Eight- and 10-mm grafts seemed better than 12-mm grafts, and grafts should be kept shorter than 25 mm. Simulation of CFD may shed light on the issue of the optimal intercostal artery reconstruction technique. [ABSTRACT FROM AUTHOR]

Open in new tab Download slide OBJECTIVES 18-Fluorodeoxyglucose (18F-FDG) positron emission tomography (PET)/computed tomography (CT) has been reported as useful for diagnosing aortic graft infection. However, 18F-FDG uptake may depend upon various factors including open versus endovascular repair and time from surgery. We aimed to elucidate the factors influencing its uptake and the diagnostic value of 18F-FDG PET/CT after open and endovascular repair. METHODS Hospital database of PET/CT (N  = 14 490) and our departmental database were cross-checked to identify those who underwent 18F-FDG PET/CT after aortic repair. Patient's data were retrieved from the chart. Images were reviewed by 2 nuclear medicine specialists in consensus, and the presence of increased 18F-FDG uptake was recorded. The maximum standardized uptake value (SUV max) was measured. RESULTS Among the 1112 patients who underwent aortic repair between 2011 and 2022, 71 patients were identified. Eighteen patients underwent 18F-FDG PET/CT for suspected graft infection and the remaining 53 patients for other purposes (malignancy, etc.). Fourteen patients were treated as aortic graft infection. They had significantly higher SUV max than those without graft infection [mean 8.64 (standard deviation 2.78) vs 3.40 (standard deviation 0.84); P  < 0.01]. In the non-infected grafts, SUV max was higher early after open surgical repair, while it remained low after endovascular repair. CONCLUSIONS After endovascular aortic repair, a constant cut-off value of 'SUV max = 4.5' seems appropriate for diagnosing graft infection, since it remains low and stable from the early postoperative period. After open surgical repair, it seems acceptable to have 'stepwise cut-off value' depending on the time from surgery. [ABSTRACT FROM AUTHOR]

Abstract: Lithospermum erythrorhizon shoots, cultured on phytohormone-free Murashige and Skoog solid medium, produced shikonin derivatives, whereas shoots cultured in well-ventilated petri dishes, produced small amount. Analysis by gas chromatography revealed the presence of ethylene in non-ventilated petri dishes where the shoots, producing shikonin derivatives, were cultured. Therefore, the possible involvement of ethylene in shikonin biosynthesis of shoot cultures was investigated. Treatment of ethylene or the ethylene precursor, 1-aminocyclopropane-1-carboxylic acid, resulted in increasing shikonin derivatives contents in cultured shoots. Silver ion, an ethylene-response inhibitor, or aminoethoxyvinylglycine, an ethylene biosynthesis inhibitor, decreased production of shikonin derivatives in cultured shoots. Our results indicate that ethylene is one of the regulatory elements of shikonin biosynthesis in L. erythrorhizon shoot culture. [Copyright &y& Elsevier]

OBJECTIVES Canine experiments have shown that transoesophageal motor-evoked potential monitoring is feasible, safe and stable, with a quicker response to ischaemia and a better prognostic value than transcranial motor-evoked potentials. We aimed to elucidate whether or not these findings were clinically reproducible. METHODS A bipolar oesophageal electrode mounted on a large-diameter silicon tube and a train of 5 biphasic wave stimuli were used for transoesophageal stimulation. Results of 18 patients (median age 74.5 years, 13 males) were analysed. RESULTS There were no mortalities, spinal cord injuries or complications related with transoesophageal stimulation. Transcranial motor-evoked potential could not be monitored up to the end of surgery in 3 patients for unknown reasons, 2 of whom from the beginning. Transoesophageal motor-evoked potential became non-evocable after manipulation of a transoesophageal echo probe in 2 patients. Strenuous movement of the upper limbs during transoesophageal stimulation was observed in 3 patients. In 14 patients who successfully completed both monitoring methods up to the end of surgery (11 thoraco-abdominal and 3 descending aortic repair), the final results were judged as false positives in 6 by transcranial stimulation and in 1 by transoesophageal stimulation. The stimulation intensity was significantly lower and the upper limb amplitude was significantly higher by transoesophageal stimulation, while the lower limb amplitude was comparable. CONCLUSIONS Transoesophageal motor-evoked potential monitoring is clinically feasible and safe with a low false positive rate. A better electrode design is required to avoid its migration by transoesophageal echo manipulation. Further studies may be warranted. Clinical registration number UMIN000022320. [ABSTRACT FROM AUTHOR]

The soil-borne bacterial pathogen Ralstonia solanacearum invades a broad range of plants through their roots, resulting in wilting of the plant, but no effective protection against this disease has been developed. Two bacterial wilt disease-inhibiting compounds were biochemically isolated from tobacco and identified as sclareol and cis-abienol, labdane-type diterpenes. When exogenously applied to their roots, sclareol and cis-abienol inhibited wilt disease in tobacco, tomato and Arabidopsis plants without exhibiting any antibacterial activity. Microarray analysis identified many sclareol-responsive genes in Arabidopsis roots, including genes encoding or with a role in ATP-binding cassette (ABC) transporters, and biosynthesis and signaling of defense-related molecules and mitogen-activated protein kinase (MAPK) cascade components. Inhibition of wilt disease by sclareol was attenuated in Arabidopsis mutants defective in the ABC transporter AtPDR12, the MAPK MPK3, and ethylene and abscisic acid signaling pathways, and also in transgenic tobacco plants with reduced expression of NtPDR1, a tobacco homolog of AtPDR12. These results suggest that multiple host factors are involved in the inhibition of bacterial wilt disease by sclareol-related compounds. [ABSTRACT FROM AUTHOR]

A pre-infestation of the white-backed planthopper (WBPH), Sogatella furcifera Horváth, conferred resistance to bacterial blight caused by Xanthomonas oryzae pv. oryzae ( Xoo) in rice ( Oryza sativa L.) under both laboratory and field conditions. The infestation of another planthopper species, the brown planthopper (BPH) Nilaparvata lugens Stål, did not significantly reduce the incidence of bacterial blight symptoms. A large-scale screening using a rice DNA microarray and quantitative RT-PCR revealed that WBPH infestation caused the upregulation of more defence-related genes than did BPH infestation. Hydroperoxide lyase 2 ( OsHPL2), an enzyme for producing C6 volatiles, was upregulated by WBPH infestation, but not by BPH infestation. One C6 volatile, ( E)-2-hexenal, accumulated in rice after WBPH infestation, but not after BPH infestation. A direct application of ( E)-2-hexenal to a liquid culture of Xoo inhibited the growth of the bacterium. Furthermore, a vapour treatment of rice plants with ( E)-2-hexenal induced resistance to bacterial blight. OsHPL2-overexpressing transgenic rice plants exhibited increased resistance to bacterial blight. Based on these data, we conclude that OsHPL2 and its derived ( E)-2-hexenal play some role in WBPH-induced resistance in rice. [ABSTRACT FROM AUTHOR]

Ethylene and jasmonic acid (JA) have been proposed as key compounds for wound signaling in plants. In Arabidopsis, ETHYLENE INSENSITIVE3 (EIN3), which is an essential transcription factor for ethylene signaling, is regulated at the post-transcriptional level, while transcriptional regulation of EIN3 or EIN3-LIKE ( EIL) genes has not been well documented. The expression of 6 rice EIL genes ( OsEIL1-6) was analyzed and only OsEIL1 and 2 were found to be wound-inducible EIL. OsEIL2 was also induced by JA. Electrophoretic mobility shift assays showed that recombinant OsEIL1 and 2 proteins bound to specific DNA sequences that are recognized by a wound-inducible tobacco EIL. Accumulation of OsEIL1 and 2 transcripts reached a maximum at 1 and 0.5 h after wounding, respectively, and the corresponding DNA-binding activity in nuclear extracts of rice leaves was increased at 1 h after wounding. Candidates for OsEIL-target genes were selected by microarray analysis of wounded rice and by promoter sequence analyses of wound-inducible genes identified by microarray analysis. In OsEIL1- and/or 2-suppressed rice plants, the expression of at least four of 18 candidate genes analyzed was down-regulated. These results indicate the importance of inducible OsEILs in wound signaling in rice. [ABSTRACT FROM AUTHOR]

The TEIL (Tobacco EIN3-Like) gene is a tobacco homologue of arabidopsis Ethylene Insensitive 3 (EIN3), and the gene product binds an 8 bp sequence in the tobacco PR1a promoter in a sequence specific manner. It was found here that accumulation of TEIL transcript was induced by wounding and preceded basic PR gene expression. To study the downstream signalling pathway of TEIL, TEIL was overexpressed under the control of the constitutive 35S promoter in tobacco plants. In 35S::TEIL lines, basic PR genes, which are wound-, jasmonate-, and ethylene-inducible, were expressed constitutively. Next, the conserved 781 bp sequence among tobacco EIN3-like (EIL) protein genes was introduced as an inverted-repeat (IR) into tobacco to suppress expression of these genes. In two independent IRTEIL lines, the TEIL transcript was not found and transcripts of other tobacco EILs, NtEIL3, and NtEIL5, were reduced. In IRTEIL plants, wound-, jasmonate-, and ACC-induced accumulation of basic PR gene transcripts was significantly inhibited. These results indicate that TEIL functions upstream of tobacco basic PR genes in wound signalling via not only ethylene but also jasmonate. In 35S::TEIL plants, the pistil length of the flower was longer with a slight protrusion of the stigma compared with the control. In IRTEIL plants, the length of the stamens was shorter than the control with significant protrusion of the stigma in the flower. These observations indicate the involvement of tobacco EILs in flower development. [ABSTRACT FROM PUBLISHER]

The vascular system-specific and wound-responsive cis-element (VWRE) has been identified as a novel cis-element for wound-induced and vascular system-specific expression of the tobacco peroxidase gene, tpoxN1. Here we isolated two independent clones that encode VWRE binding proteins by yeast one-hybrid screening. As the gene products have an AP2/ERF (APETALA2/ethylene-responsive factor) domain, and the transcripts were accumulated transiently after wounding, we named them wound-responsive AP2/ERF-like factor 1 (WRAF1) and WRAF2. The AP2/ERF domains of the two WRAFs share 97% homology, and are classified into the ERF subfamily B-4. Gel mobility shift analysis indicated that WRAFs specifically bind VWRE, which contains no known cis-elements for other AP2/ERF proteins. The binding activity of the WRAFs was found to be localized in the AP2/ERF domain. The WRAFs transactivated a promoter containing four tandem repeats of the VWRE, but not that of the mutated VWRE. Overexpression of the WRAF genes led to constitutive expression of the potential target gene, tpoxN1, in unwounded transgenic plants. These results indicate that the novel transcription factors WRAF1 and WRAF2 bind the VWRE as positive regulators for the expression of the tpoxN1 gene. [ABSTRACT FROM AUTHOR]

In tobacco ( Nicotiana tabacum), wounding causes rapid activation of two mitogen-activated protein kinases (MAPKs), wound-induced protein kinase (WIPK) and salicylic acid (SA)-induced protein kinase (SIPK), and the subsequent accumulation of jasmonic acid (JA). Our previous studies suggested that activation of WIPK is required for the production of wound-induced JA. However, the exact role of WIPK remains unresolved. We generated transgenic tobacco plants in which either WIPK or SIPK were silenced using RNA interference to define the roles of WIPK and SIPK in the wound response. In addition, transgenic tobacco plants were generated in which both WIPK and SIPK were silenced to examine the possibility that they have redundant roles. Wound-induced JA production was reduced compared with non-silenced plants in all of the WIPK-, SIPK- and WIPK/SIPK-silenced plants. Transgenic plants over-expressing NtMKP1, a gene encoding tobacco MAPK phosphatase, which inactivates WIPK and SIPK, also exhibited reduced JA production in response to wounding. In both WIPK/SIPK-silenced and NtMKP1-over-expressing plants, wounding resulted in an abnormal accumulation of both SA and transcripts for SA-responsive genes. These results suggest that WIPK and SIPK play an important role in JA production in response to wounding, and that they function cooperatively to control SA biosynthesis. [ABSTRACT FROM AUTHOR]

The role of ethylene (ET) in resistance to infection with blast fungus (Magnaporthe grisea) in rice (Oryza sativa) is poorly understood. To study it, we quantified ET levels after inoculation, using young rice plants at the four-leaf stage of rice cv Nipponbare (wild type) and its isogenic plant (IL7), which contains the Pi-i resistance gene to blast fungus race 003. Small necrotic lesions by hypersensitive reaction (HR) were formed at 42 to 72 h postinoculation (hpi) in resistant IL7 leaves, and whitish expanding lesions at 96 hpi in susceptible wild-type leaves. Notable was the enhanced ET emission at 48 hpi accompanied by increased 1-aminocyclopropane-1-carboxylic acid (ACC) levels and highly elevated ACC oxidase (ACO) activity in IL7 leaves, whereas only an enhanced ACC increase at 96 hpi in wild-type leaves. Among six ACC synthase (ACS) and seven ACO genes found in the rice genome, OsACS2 was transiently expressed at 48 hpi in IL7 and at 96 hpi in wild type, and OsACO7 was expressed at 48 hpi in IL7. Treatment with an inhibitor for ACS, aminooxyacetic acid, suppressed enhanced ET emission at 48 hpi in IL7, resulting in expanding lesions instead of HR lesions. Exogenously supplied ACC compromised the aminooxyacetic acid-induced breakdown of resistance in IL7, and treatment with 1-methylcyclopropene and silver thiosulfate, inhibitors of ET action, did not suppress resistance. These findings suggest the importance of ET biosynthesis and, consequently, the coproduct, cyanide, for HR-accompanied resistance to blast fungus in young rice plants and the contribution of induced OsACS2 and OsACO7 gene expression to it. [ABSTRACT FROM AUTHOR]

The wound-induced receptor-like protein kinase ( WRK) gene, isolated as one of the genes whose transcripts accumulated during the early period of N gene-dependent synchronized cell death in tobacco mosaic virus-infected tobacco plants, encodes a leucine-rich repeat receptor-like protein kinase, and its transcript was transiently increased 15 min after wounding. In the present study, analysis of a green fluorescent protein fusion protein indicated that WRK is localized in the plasma membrane. In transgenic tobacco plant lines with elevated or suppressed levels of WRK transcript, the wound-induced accumulation of both basic PR-1 and PR-6 transcripts was slightly enhanced or significantly suppressed respectively. The decrease in wound-induced basic PR gene expression in WRK suppressed lines was restored by jasmonic acid (JA) treatment. Furthermore, the levels of wound-induced enzymatic activation of both salicylic acid-induced protein kinase (SIPK) and wound-induced protein kinase (WIPK) and wound-induced accumulation of JA were reduced in the WRK suppressed lines in comparison with a control line. These results suggest that WRK functions upstream of SIPK and WIPK and regulates wound signal transduction in tobacco plants. [ABSTRACT FROM AUTHOR]

Abstract: Expression of defense-related genes was induced within 1h after blast fungus inoculation in both resistant and susceptible rice plants, on which the conidia on the leaves have just germinated. Treatment of rice cells with water-soluble and heat-sensitive molecule(s) released from the conidia induced rapid defense gene expression, but not the rapid cytoplasmic pH decrease, which was reportedly induced by elicitors originating from blast fungus mycelium. These results suggest that, before penetration into plant cells, a rapid defensive response to blast fungus has been triggered by conidia-originating elicitor(s) in both resistant and susceptible rice plants. [Copyright &y& Elsevier]

MAPK signal transduction modules play crucial roles in regulating many biological processes in plants, and their components are encoded by highly conserved genes. The recent availability of genome sequences for rice and poplar now makes it possible to examine how well the previously described Arabidopsis MAPK and MAPKK gene family structures represent the broader evolutionary situation in plants, and analysis of gene expression data for MPK and MKK genes in all three species allows further refinement of those families, based on functionality. The Arabidopsis MAPK nomenclature appears sufficiently robust to allow it to be usefully extended to other well-characterized plant systems. [Copyright &y& Elsevier]

The N gene is a Toll/interleukin-1 receptor (TIR)–nucleotide-binding site (NBS)–leucine-rich repeat (LRR)-type resistance (R) gene that generates two alternative transcripts, NS and NL. NS encodes the full-length N protein while NL is predicted to encode a truncated form of the protein lacking most of the LRR region. We found that the two transcripts were accumulated at 20°C, a permissive temperature, but not at 30°C, a non-permissive temperature for the N gene, in tobacco mosaic virus (TMV)-inoculated leaves. When N gene-dependent cell death was triggered by transient 20°C treatment for 2–6 h, considerable levels of the transcripts were accumulated just before cell death, although the levels of NS were always higher. The accumulation was induced by transient expression of the 50 kDa helicase domain (p50) of TMV replicase which is the Avr component of N, but not by transient expression of NtMEK2DD-mediated cell death or N gene-independent hypersensitive cell death. These results suggest that the accumulation of NS and NL is associated with the function of N and, above a certain threshold, triggers N-mediated hypersensitive cell death. [ABSTRACT FROM AUTHOR]

The mitogen-activated protein kinase (MAPK) cascade is involved in responses to biotic and abiotic stress in plants. In this study, we isolated a new MAPK, NtMPK4, which is a tobacco homolog of Arabidopsis MPK4 (AtMPK4). NtMPK4 was activated by wounding along with two other wound-responsive tobacco MAPKs, WIPK and SIPK. We found that NtMPK4 was activated by salicylic acid-induced protein kinase kinase (SIPKK), which has been isolated as an SIPK-interacting MAPK kinase. In NtMPK4 activity-suppressed tobacco, wound-induced expression of jasmonic acid (JA)-responsive genes was inhibited. NtMPK4-silenced plants showed enhanced sensitivity to ozone. Inversely, transgenic tobacco plants, in which SIPKK or the constitutively active type SIPKKEE was overexpressed, exhibited greater responsiveness to wounding with enhanced resistance to ozone. We further found that NtMPK4 was expressed preferentially in epidermis, and the enhanced sensitivity to ozone in NtMPK4-silenced plants was caused by an abnormal regulation of stomatal closure in an ABA-independent manner. These results suggest that NtMPK4 is involved in JA signaling and in stomatal movement. [ABSTRACT FROM AUTHOR]

MAPK phosphatases (MKPs) are negative regulators of MAPKs. Previously, we identified NtMKP1 as a novel calmodulin (CaM)-binding protein (Yamakawa, H., Katou, S., Seo, S., Mitsuhara, I., Kamada, H., and Ohashi, Y. (2004) J. Biol. Chem. 279, 928–936). In this study, we characterized the interaction of NtMKP1 with substrate MAPKs and CaM. NtMKP1 (produced by in vitro transcription/translation) inactivated salicylic acid-induced protein kinase (SIPK) through dephosphorylation of the TEY motif of SIPK. CaM bound but unexpectedly did not activate the phosphatase activity of NtMKP1. NtMKP1 has four characteristic domains, viz. a dual-specificity phosphatase catalytic domain, a gelsolin homology domain, a CaM-binding domain, and C-terminal domain. Deletion analysis revealed that the N-terminal non-catalytic region of NtMKP1 bound SIPK and was essential for inactivating SIPK, whereas the CaM-binding and C-terminal domains were dispensable. Moreover, the phosphatase activity of NtMKP1 was increased strongly by the binding of SIPK, but weakly by another MAPK, wound-induced protein kinase. Swapping and site-directed mutagenesis of SIPK and wound-induced protein kinase revealed that the strong activation of NtMKP1 phosphatase activity by SIPK partially depended on the putative common docking domain of SIPK. On the other hand, conversion of Lys41 and Arg43 of NtMKP1 to Ala (K41A/R43A) abolished the interaction with SIPK. Expression of constitutively active MAPK kinase in Nicotiana benthamiana induced activation of SIPK and cell death. Simultaneous expression of either NtMKP1 or NtMKP1 L443R, which was unable to bind CaM, compromised the constitutively active MAPK kinase-induced responses, whereas that of NtMKP1 K41A/R43A did not. These results indicate that the regulation of NtMKP1 activity by SIPK binding, but not by CaM binding, is important for the function of NtMKP1. [ABSTRACT FROM AUTHOR]

We previously reported that three types of tobacco calmodulin (CaM) isoforms originated from 13 genes are differently regulated at the transcript and protein levels in response to wounding and tobacco mosaic virus-induced hypersensitive reaction (HR); wound-inducible type I and HR-inducible type III levels increased after wounding and HR, respectively, while type II, whose expression is constitutive and wound responsible, remained unchanged. Here, we show that these CaMs differentially activate target enzymes; rat NO synthase was activated most effectively by type III, moderately by type I and weakly by type II, and plant NAD kinase (NADK) was activated in the inverse order. Furthermore, we found that a suitable Ca2+ concentration differs by type; type II activated NADK at lower Ca2+ of around 0.1 µM, which is the cytosolic concentration in unstimulated cells, type I did so at 1–5 µM, which is the increased Ca2+ concentration in stimulated cells, while type III did not at any Ca2+ level. NADK activation was highest over a pH range of 7.1–6.8 for which the cytosolic pH reportedly changed from 7.5 after being stimulated. Thus, tobacco CaMs, especially type I, effectively activate NADK in stimuli-induced conditions. [ABSTRACT FROM AUTHOR]

Class III plant peroxidases are believed to function in diverse physiological processes including disease resistance and wound response, but predicted low substrate specificities and the presence of 70 or more isoforms have made it difficult to define a specific physiological function(s) for each gene. To select pathogen-responsive POX genes, we analyzed the expression profiles of 22 rice POX genes after infection with rice blast fungus. The expression of 10 POX genes among the 22 genes was induced after fungal inoculation in both compatible and incompatible hosts. Seven of the 10 POX genes were expressed at higher levels in the incompatible host than in the compatible host 6–24 h after inoculation by which time no fungus-induced lesions have appeared. Organ-specific expression and stress-induced expression by wounding and treatment with probenazole, an agrichemical against blast fungus, jasmonic acid, salicylic acid and 1-aminocyclopropane-1-carboxylate, a precursor of ethylene, indicated that rice POXs have individual characteristics and can be classified into several types. A comparison of the amino acid sequences of POXs showed that multiple isoforms with a high sequence similarity respond to stress in different or similar ways. Such redundant responses of POX genes may guarantee POX activities that are necessary for self-defense in plant tissues against environmental stresses including pathogen infection. [ABSTRACT FROM AUTHOR]

Describes the mechanisms by which plant MAPK phosphatase interacts with calmodulins. Dual specificity protein phosphatase catalytic domain; Mosaic virus-induced hypersensitive reaction; Binding proteins.

Polyamines (PAs) play important roles in cell proliferation, growth and environmental stress responses of all living organisms. In this study, we examine whether these compounds act as signal mediators. Spermine (Spm) specifically activated protein kinases of tobacco leaves, which were identified as salicylic acid (SA)-induced protein kinase (SIPK) and wound-induced protein kinase (WIPK), using specific antibodies. Upon Spm treatment, upregulation of WIPK, but not SIPK, was observed. Spm-induced mitogen-activated protein kinases (MAPKs) activation and WIPK upregulation were prevented upon pre-treatment with antioxidants and Ca2+ channel blockers. Additionally, Spm specifically stimulated expression of the alternative oxidase ( AOX) gene, which was disrupted by these antioxidants and Ca2+ channel blockers. Bongkrekic acid (BK), an inhibitor of the opening of mitochondrial permeability transition (PT) pores, suppressed MAPKs activation and accumulation of WIPK and AOX mRNA. Our data collectively suggest that Spm causes mitochondrial dysfunction via a signalling pathway in which reactive oxygen species and Ca2+ influx are involved. As a result, the phosphorylation activities of the two MAPK enzymes SIPK and WIPK are stimulated. [ABSTRACT FROM AUTHOR]

Examines the isolation and chemical identification of a wound-induced protein kinase (WIPK)-activating substance from tobacco leaves. Response to tobacco mosaic virus; Treatment of leaves with WAF-1; Role in the induction of WIPK and wound- and pathogen-inducible pathogenesis-related genes in wounded tobacco plants.

Overproduction of animal cell death suppressors Bcl-xL and Ced-9 conferred enhanced resistance to UV-B and paraquat treatment in tobacco plants [Mitsuhara et al. (1999) Curr. Biol. 9: 775]. We report here that the progeny could germinate in 0.2 M NaCl or at 10°C under light, while control plants could not. Suspension-cultured Bcl cells resisted NaCl treatment maintaining an active mitochondrial membrane potential for longer than control cells. When intracellular pH was determined by in vivo 31P-NMR, immediate cytoplasmic acidification by 0.3 M NaCl treatment in control cells was found to be suppressed in transgenic cells. Monitoring of cytoplasmic and vacuolar pHs in control cells indicated the vacuole was disrupted 40 min after 0.5 M NaCl treatment, while the compartment between the cytoplasm and vacuole was likely to remain intact in Bcl cells for 100 min. Enhanced shoot regeneration from cut leaf pieces and more vigorous rooting from cut stem ends were found in transgenic plants. The Bcl protein was abundant in all subcellular fractions. Based on the results in transgenic plants carrying a mutant bcl-xL gene, Bcl-xL is thought to suppress cell death and enhance the viability of plants in stressful environments by contributing to the maintenance of the homeostasis of organella. [ABSTRACT FROM PUBLISHER]

Mitogen-activated protein kinase (MAPK) cascades are universal signal transduction modules in eukaryotes, including yeasts, animals and plants. These protein phosphorylation cascades link extracellular stimuli to a wide range of cellular responses. In plants, MAPK cascades are involved in responses to various biotic and abiotic stresses, hormones, cell division and developmental processes. Completion of the Arabidopsis genome-sequencing project has revealed the existence of 20 MAPKs, 10 MAPK kinases and 60 MAPK kinase kinases. Here, we propose a simplified nomenclature for Arabidopsis MAPKs and MAPK kinases that might also serve as a basis for standard annotation of these gene families in all plants. [ABSTRACT FROM AUTHOR]

A gene encoding a receptor-like protein kinase was isolated as the gene induced in the early period of N gene-dependent hypersensitive cell death in tobacco leaves. The kinase domain expressed as a glutathione S-transferase fusion protein was capable of autophosphorylation, indicating that this gene encodes an active protein kinase. A high level of the transcript accumulated before necrotic lesion formation in tobacco mosaic virus (TMV)-inoculated tobacco leaves carrying the N gene but it was low in a tobacco cultivar lacking the N gene. A small but reproducible increase in the transcript was found 1–2 h after a temperature shift from 30°C to 20°C even in healthy leaves, suggesting the gene expression is temperature sensitive. The gene was named WRK for wound-induced receptor-like protein kinase, because the transcript increased to a maximum within 15–30 min of wounding. In suspension cultured tobacco cells, an increase in the transcript was found 15 min after transfer to a new medium, but it was suppressed under high osmotic pressures. The wound-induced WRK accumulation was enhanced by cycloheximide treatment, but not by known defense signal compounds (salicylic acid, jasmonic acid, 1-aminocyclopropan-1-carboxylic acid and abscisic acid) and some plant hormones. Thus, WRK is a wound-inducible and temperature-sensitive protein kinase gene induced before hypersensitive cell death probably through unknown signaling pathways. [ABSTRACT FROM PUBLISHER]

A tobacco peroxidase gene tpoxN1 was reported to be expressed within 1 h after wounding in leaves [Hiraga et al. (2000a) Plant Cell Physiol. 41: 165]. We describe here further results on the wound-induced tpoxN1 expression. The quick tpoxN1 induction occurred preferentially in stems and petioles, but was negligible in leaf blades even 8 h after wounding. Induced GUS activity was also detected rapidly after wounding in the stem of transgenic tobacco plants carrying the tpoxN1 promoter::GUS fusion gene, localized mainly in the vascular systems where it was maintained this level for 14 d or more. Strong GUS activity was also found in the petiole and veinlet as well as the epidermal tissue in the stem. Treatment of known inducers for wound-responsive genes such as jasmonate, 1-aminocyclopropane-1-carboxylate, spermine, phytohormones and other stress treatments did not enhance wound-induced tpoxN1 gene expression in stems at all, but rather repressed it in some cases. Studies using metabolic inhibitors suggested that phosphorylation and dephosphorylation of proteins together with de novo protein synthesis are likely to be involved in the wound-induced tpoxN1 expression as well as some other wound-responsive genes. Thus, tpoxN1 is a unique wound-inducible and possible wound-healing gene which is rapidly expressed being maintained for a long time in veins via an unknown wound-signaling pathway(s). [ABSTRACT FROM PUBLISHER]

Discusses the release from post-transcriptional gene silencing by cell proliferation in transgenic tobacco plants. Transgene silencing in tobacco plants overproducing a firefly luciferase gene; Biochemical analysis.

We isolated 13 tobacco calmodulin (CaM) genes, NtCaM1–13, and analyzed their expression profile in response to pathogen infection and wounding using specific DNA probes for individual CaM genes and specific antibodies for CaM proteins in groups I (NtCaM1/2), II (NtCaM3/4/5/6/7/8/11/12 and 9/10) and III (NtCaM13), respectively. Synchronous cell death in tobacco mosaic virus (TMV)-infected N-gene-containing tobacco leaves accompanied a predominant accumulation of NtCaM1, 2 and 13 transcripts and NtCaM13-type protein, which is a possible ortholog of soybean defense-involved CaM (SCaM-4), preceding induction of PR-1 and PR-3 defense genes. Accumulation of NtCaM1, 2, 3 and 4 transcripts was induced within 30 min after wounding and NtCaM1-type protein accumulated transiently after wounding. NtCaM13-type protein, which was found at a low level in healthy leaves, decreased instantly after wounding. The treatment with a proteasome inhibitor, lactacystin, enhanced wound-induced accumulation of NtCaM1-type protein and inhibited wound-induced decrease of NtCaM13-type protein, suggesting that proteasome activity is involved in the degradation of these CaMs. Thus, our results indicate that levels of individual CaM proteins are differentially regulated both transcriptionally and post-transcriptionally in tobacco plants that are exposed to stresses such as pathogen-induced hypersensitive cell death and wounding. [ABSTRACT FROM AUTHOR]

Class III plant peroxidase (POX), a plant-specific oxidoreductase, is one of the many types of peroxidases that are widely distributed in animals, plants and microorganisms. POXs exist as isoenzymes in individual plant species, and each isoenzyme has variable amino acid sequences and shows diverse expression profiles, suggesting their involvement in various physiological processes. Indeed, studies have provided evidence that POXs participate in lignification, suberization, auxin catabolism, wound healing and defense against pathogen infection. Little, however, is known about the signal transduction for inducing expression of the pox genes. Recent studies have provided information on the regulatory mechanisms of wound- and pathogen-induced expression of some pox genes. These studies suggest that pox genes are inducedviadifferent signal transduction pathways from those of other known defense-related genes. [ABSTRACT FROM PUBLISHER]

Objectives: Although cerebrospinal fluid drainage has been shown to reduce the risk of ischemic spinal cord injury, serious complications have also been reported. We have been using it selectively in a pressure- and volume-regulated method and aimed to evaluate its safety, and its validity in elective thoracic endovascular aortic repair in a propensity-matched cohort., Methods: Among the 450 patients who underwent open surgery (n = 169) or thoracic endovascular aortic repair (n = 281) on the descending or thoracoabdominal aorta, 147 underwent cerebrospinal fluid drainage, which was prophylactic in 135 and therapeutic in 12. Prophylactic drainage was performed in elective open surgery under distal aortic perfusion (n = 67) or in selected patients undergoing thoracic endovascular aortic repair (n = 68)., Results: Drainage-related complications were observed in 13 (9.6%), one of which was graded severe (0.74%). In patients undergoing prophylactic drainage, spinal cord injury was detected in 2/135 (1.5%). In patients without prophylactic drainage, 15/315 (4.8%) developed spinal cord injury. Therapeutic drainage was performed in 12 of these 15 patients, 10 of whom remained paralytic in varying degree. In the inverse probability weighted analysis of the patients undergoing elective thoracic endovascular aortic repair, the incidence of spinal cord injury was lower with prophylactic drainage (p = 0.028)., Conclusions: Pressure- and volume-regulated spinal drainage rarely causes serious complications. Its prophylactic use seems beneficial in selected patients, including those undergoing thoracic endovascular aortic repair with high risk for spinal cord injury., (© 2024. The Author(s).)

Objective: In the loop technique for mitral valve repair, the loop bundles are usually created during cardiac arrest after chordal length measurements, which seems time-consuming and less reproducible. To address this issue, we determined the loop length preoperatively using 4-dimensional computed tomography., Methods: The loop length was determined on the basis of the distance from the papillary muscle head to the free margin of nonprolapsing leaflet corresponding to the prolapsed leaflet, to which the loops would be secured. Measurements were made on the commissural and long-axis views created by a medical image post-processor in the late systolic phase. This technique was used in consecutive 45 patients undergoing mitral valve repair with the loop technique since April 2021., Results: A total of 55 loop bundles were created in 45 patients; in 10 cases loop bundles were fixed to both anterior and posterior papillary muscles. There were 31 posterior, 6 anterior, and 8 bileaflet prolapse. The loop length was set at 16 to 26 mm (median 19 mm). Mitral valve repair was successfully completed in all patients, and the loop bundles of predetermined length were used successfully in 42 patients (93.3%). Postoperative echocardiography revealed none/trace regurgitation in 41 and mild regurgitation in 4. There was no hospital mortality or major postoperative complication. During 4 to 35 months follow-up (median 10 months), no case required reintervention for the mitral valve., Conclusions: Preoperative measurements using 4-dimensional computed tomography can accurately and reproducibly predict the required loop length for mitral valve repair., Competing Interests: The authors reported no conflicts of interest. The Journal policy requires editors and reviewers to disclose conflicts of interest and to decline handling or reviewing manuscripts for which they may have a conflict of interest. The editors and reviewers of this article have no conflicts of interest., (© 2024 The Author(s).)