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1.
Circ J ; 84(7): 1090-1096, 2020 06 25.
Artigo em Inglês | MEDLINE | ID: mdl-32461539

RESUMO

BACKGROUND: Bridge-to-decision (BTD) devices providing temporary mechanical circulatory support should be introduced to patients with advanced heart failure. This study evaluated the effectiveness and safety of a BTD device comprising an innovative extracorporeal continuous-flow temporary ventricular assist device (VAD) driven by a novel hydrodynamically levitated centrifugal flow blood pump.Methods and Results:Nine patients, comprising 3 with dilated cardiomyopathy, 3 with fulminant myocarditis, and 3 with ischemic heart disease, and 6 males, whose mean age was 47.7±8.1 years, were enrolled into the study. Six patients had Interagency Registry for Mechanically Assisted Circulatory Support profile 1, and 3 were profile 2. The primary endpoint was a composite of survival free from device-related serious adverse events and complications during circulatory support. Eight patients received left ventricular support, of whom 3 received concomitant right ventricular support using extracorporeal membrane oxygenation circuits, as a consequence of severe respiratory failure. One patient with fulminant myocarditis received biventricular support using the novel VAD system. After 19.0±13.5 days, 3 patients were weaned from circulatory support, because their native cardiac function recovered, and 6 patients required conversion to a durable device as a bridge-to-transplantation. One patient had non-disabling ischemic stroke episodes, and no patients died. CONCLUSIONS: This novel extracorporeal VAD system with a hydrodynamically levitated centrifugal pump can safely and successfully bridge patients with advanced heart failure to subsequent therapeutic stages.


Assuntos
Oxigenação por Membrana Extracorpórea/instrumentação , Insuficiência Cardíaca/terapia , Coração Auxiliar , Implantação de Prótese/instrumentação , Função Ventricular Esquerda , Função Ventricular Direita , Adulto , Oxigenação por Membrana Extracorpórea/efeitos adversos , Oxigenação por Membrana Extracorpórea/mortalidade , Feminino , Insuficiência Cardíaca/diagnóstico , Insuficiência Cardíaca/mortalidade , Insuficiência Cardíaca/fisiopatologia , Transplante de Coração , Humanos , Masculino , Pessoa de Meia-Idade , Desenho de Prótese , Implantação de Prótese/efeitos adversos , Implantação de Prótese/mortalidade , Recuperação de Função Fisiológica , Fatores de Tempo , Resultado do Tratamento
2.
Cardiovasc Drugs Ther ; 32(4): 373-379, 2018 08.
Artigo em Inglês | MEDLINE | ID: mdl-29948739

RESUMO

BACKGROUND: The management of heart failure patients presenting in a moribund state remains challenging, despite significant advances in the field of ventricular assist systems. Bridge to decision involves using temporary devices to stabilize the hemodynamic state of such patients while further assessment is performed and a decision can be made regarding patient management. The purpose of this study (NCVC-BTD_01, National Cerebral and Cardiovascular Center-Bridge to Dicision_01) is to assess the safety and effectiveness of the newly developed extracorporeal continuous-flow ventricular assist system employing a disposable centrifugal pump with a hydrodynamically levitated bearing (BR16010) use as a bridge-to-decision therapy for patients with severe heart failure or refractory cardiogenic shock. METHOD/DESIGN: NCVC-BTD_01 is a single-center, single-arm, open-label, exploratory, medical device, investigator-initiated clinical study. It is conducted at the National Cerebral and Cardiovascular Center in Japan. A total of nine patients will be enrolled in the study. The study was planned using Simon's minimax two-stage phase design. The primary endpoint is a composite of survival free of device-related serious adverse events and complications during device support. For left ventricular assistance, withdrawal of a trial device due to cardiac function recovery or exchange to other ventricular assist devices (VADs) for the purpose of bridge to transplantation (BTT) during 30 days after implantation will be considered study successes. For right ventricular assistance, withdrawal of tal device due to right ventricular function recovery within 30 days after implantation will be considered a study success. Secondary objectives include changes in brain natriuretic peptide levels (7 days after implantation of a trial device and the day of withdrawal of a trial device), period of mechanical ventricular support, changes in left ventricular ejection fraction (7 days after implantation of a trial device and the day of withdrawal of a trial device), and changes in left ventricular diastolic dimension (7 days after implantation of a trial device and the day of withdrawal of a trial device). ETHICS AND DISSEMINATION: We will disseminate the findings through regional, national, and international conferences and through peer-reviewed journals. TRIAL REGISTRATION: UMIN Clinical Trials Registry (UMIN-CTR; R000033243) registered on 8 September 2017.


Assuntos
Oxigenação por Membrana Extracorpórea/instrumentação , Insuficiência Cardíaca/terapia , Coração Auxiliar , Choque Cardiogênico/terapia , Oxigenação por Membrana Extracorpórea/efeitos adversos , Insuficiência Cardíaca/diagnóstico , Insuficiência Cardíaca/fisiopatologia , Transplante de Coração , Hemodinâmica , Humanos , Japão , Desenho de Prótese , Recuperação de Função Fisiológica , Índice de Gravidade de Doença , Choque Cardiogênico/diagnóstico , Choque Cardiogênico/fisiopatologia , Fatores de Tempo , Resultado do Tratamento , Função Ventricular Esquerda , Função Ventricular Direita
3.
Angew Chem Int Ed Engl ; 57(19): 5413-5417, 2018 05 04.
Artigo em Inglês | MEDLINE | ID: mdl-29537622

RESUMO

A cationic fluorescent nanogel thermometer based on thermo-responsive N-isopropylacrylamide and environment-sensitive benzothiadiazole was developed with a new azo compound bearing imidazolium rings as the first cationic radical initiator. This cationic fluorescent nanogel thermometer showed an excellent ability to enter live mammalian cells in a short incubation period (10 min), a high sensitivity to temperature variations in live cells (temperature resolution of 0.02-0.84 °C in the range 20-40 °C), and remarkable non-cytotoxicity, which permitted ordinary cell proliferation and even differentiation of primary cultured cells.

4.
J Artif Organs ; 19(2): 196-9, 2016 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-26670317

RESUMO

A 4-year-old boy with atrioventricular discordance, double-outlet right ventricle, pulmonary stenosis, and mitral regurgitation, was undergoing anatomical repair consisting of Senning, Rastelli, Damus-Kaye-Stansel procedures, and a mitral valve repair, complained of post-operative excessive airway tract secretion, which ultimately developed into acute respiratory distress syndrome (ARDS) 28 days after the operation. The cause of the ARDS was thought to be frequent manual positive pressure recruitment and prolonged inhalation of pure oxygen. At 45 days after the operation, hypercapnia and respiratory acidosis turned out to be irreversible, and therefore, veno-arterial extracorporeal membrane oxygenation (ECMO) was established utilizing the Endumo(®)4000 system. Pulmonic interstitial inflammation gradually improved while resting the lung under ECMO support; however, effective ventilation volume decreased critically because a massive pulmonary hemorrhage occurred at 2 and 9 days after the initiation of ECMO. To maximize the effectiveness of respiratory physical therapy, "Awake ECMO" was started and tidal volume dramatically increased with a regained cough reflex. Five days later, he was successfully weaned off from ECMO, and discharged 7 months after the operation without any neurological and physiological sequelae.


Assuntos
Procedimentos Cirúrgicos Cardíacos , Oxigenação por Membrana Extracorpórea/métodos , Complicações Pós-Operatórias/terapia , Síndrome do Desconforto Respiratório/terapia , Pré-Escolar , Dupla Via de Saída do Ventrículo Direito/cirurgia , Humanos , Masculino , Insuficiência Respiratória , Volume de Ventilação Pulmonar
5.
Plant J ; 77(1): 146-59, 2014 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-24329948

RESUMO

For the establishment of an effective root nodule symbiosis, a coordinated regulation of the infection processes between the epidermis and cortex is required. However, it remains unclear whether the symbiotic genes identified so far are involved in epidermal and/or cortical infection, e.g. epidermal and cortical infection thread formation or cortical cell division. To analyze the symbiotic gene requirements of the infection process, we have developed an epidermis-specific expression system (pEpi expression system) and examined the symbiotic genes NFR1, NFR5, NUP85, NUP133, CASTOR, POLLUX, CCaMK, CYCLOPS, NSP1 and NSP2 for involvement in the infection process in the epidermis and cortex. Our study shows that expression of the upstream common symbiosis genes CASTOR, POLLUX, NUP85 and NUP133 in the epidermis is sufficient to induce formation of infection threads and cortical cell division, leading to the development of fully effective nodules. Our system also shows a requirement of CCaMK, CYCLOPS, NSP1 and NSP2 for the entire nodulation process, and the different contributions of NFR1 and NFR5 to cortical infection thread formation. Based on these analyses using the pEpi expression system, we propose a functional model of symbiotic genes for epidermal and cortical infection.


Assuntos
Cálcio/metabolismo , Regulação da Expressão Gênica de Plantas , Lotus/genética , Rhizobium/genética , Simbiose/genética , Divisão Celular , Genes Reporter , Vetores Genéticos , Lotus/citologia , Lotus/microbiologia , Lotus/fisiologia , Modelos Biológicos , Mutação , Motivos de Nucleotídeos , Especificidade de Órgãos , Fenótipo , Epiderme Vegetal , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Nodulação , Raízes de Plantas/metabolismo , Regiões Promotoras Genéticas/genética , Rhizobium/fisiologia , Nódulos Radiculares de Plantas/genética
6.
Analyst ; 140(13): 4498-506, 2015 Jul 07.
Artigo em Inglês | MEDLINE | ID: mdl-25988198

RESUMO

We developed new cationic fluorescent polymeric thermometers containing both benzothiadiazole and BODIPY units as an environment-sensitive fluorophore and as a reference fluorophore, respectively. The temperature-dependent fluorescence spectra of the thermometers enabled us to perform highly sensitive and practical ratiometric temperature sensing inside living mammalian cells. Intracellular temperatures of non-adherent MOLT-4 (human acute lymphoblastic leukaemia) and adherent HEK293T (human embryonic kidney) cells could be monitored with high temperature resolutions (0.01-1.0 °C) using the new cationic fluorescent polymeric thermometer.


Assuntos
Compostos de Boro/química , Corantes Fluorescentes/química , Espaço Intracelular/química , Polímeros/química , Termômetros , Linhagem Celular Tumoral , Células HEK293 , Humanos , Espaço Intracelular/fisiologia , Espectrofotometria Ultravioleta/métodos , Temperatura
7.
J Artif Organs ; 18(4): 370-2, 2015 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-25854605

RESUMO

Cold agglutination was suspected in 2 pediatric open heart surgery cases during mild hypothermic cardiopulmonary bypass. The first patient was a 2-year-old boy with secundum atrial septal defect. Fifteen minutes after the initiation of mild hypothermic cardiopulmonary bypass, the inlet pressure of oxygenator suddenly elevated from 250 to over 500 mmHg, whereas outlet pressure was maintained. The blood flow rate decreased from 140 to 85 ml/kg/min. At that time, the arterial blood temperature was less than 32°C. Cold agglutinin was highly suspected, so patient was immediately warmed, and the inlet pressure of oxygenator decreased to 250 mmHg when the arterial blood temperature reached to 36°C. Second patient was a 3-year-old boy with secondary developed subvalvular pulmonary stenosis after the repair of double chambered right ventricle at 10 months of his age. Eighteen minutes after the initiation of mild hypothermic cardiopulmonary bypass, the inflow pressure suddenly elevated to 500 mmHg and transmission flow decreased to 55 ml/kg/min. Twenty-three minutes after warming, the pressure fell to a normal level and transmission flow was recovered. The operation continued with normo-thermic cardiopulmonary bypass and crystalloid cardioplegia. Both cases had no postoperative complications related to cold agglutinin such as myocardial infarction, cerebral infarction, or renal insufficiency.


Assuntos
Ponte Cardiopulmonar/efeitos adversos , Agregação Eritrocítica , Parada Cardíaca Induzida/efeitos adversos , Cardiopatias Congênitas/cirurgia , Hipotermia Induzida/efeitos adversos , Pré-Escolar , Soluções Cristaloides , Humanos , Soluções Isotônicas , Masculino
8.
J Artif Organs ; 17(4): 315-20, 2014 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-25119706

RESUMO

This study aimed to assess the efficacy of a new pediatric extra-corporeal life support (ECLS) system (Endumo 2000, Heiwa Bussan, Tokyo, Japan) for postoperative management after the Norwood operation. Thirty-three consecutive patients with hypoplastic left heart syndrome or its variant undergoing the Norwood operation between August 2007 and December 2013 were divided into two groups according to available ECLS devices. Before November 2009, pediatric Emersave (TERUMO, Saitama, Japan) was employed as the ECLS device, and 14 patients were operated on during this period (Emersave era: 7 boys; 2.9 kg). After December 2009, Endumo 2000 was employed and 19 patients were operated on (Endumo era: 8 boys, 3.1 kg). The demographic characteristics of both groups showed no significant differences. ECLS was initiated in 7 of 14 patients (50%) during the Emersave era and 7 of 19 patients (37%) during the Endumo era (p = 0.45). Chest reentry for hemostasis during ECLS support was more frequently needed in patients supported by Emersave (5/7) than Endumo (1/7) (p = 0.03). The first ECLS circuit durability of Endumo was significantly longer than that of Emersave (p = 0.01). The survival at discharge rate in patients required ECLS was 0% (0/7) when supported by Emersave, but 57% (4/7) by Endumo (p = 0.02). As a result, the survival at discharge rate was 43% (6/14) in the Emersave era and 79% (14/19) in the Endumo era (p = 0.03). Longer durability and superior antithrombogenicity of the Endumo 2000 contributed to the improvement of surgical outcomes after the Norwood operation.


Assuntos
Oxigenação por Membrana Extracorpórea/instrumentação , Síndrome do Coração Esquerdo Hipoplásico/cirurgia , Sistemas de Manutenção da Vida/instrumentação , Procedimentos de Norwood/métodos , Oxigenação por Membrana Extracorpórea/mortalidade , Feminino , Humanos , Síndrome do Coração Esquerdo Hipoplásico/mortalidade , Lactente , Recém-Nascido , Masculino , Procedimentos de Norwood/mortalidade , Complicações Pós-Operatórias/mortalidade , Resultado do Tratamento
9.
J Artif Organs ; 17(1): 99-102, 2014 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-24379041

RESUMO

A 5-year-old girl with right atrial isomerism, complete atrioventricular septal defect, hypoplastic left ventricle, double outlet right ventricle, and mixed-type total anomalous pulmonary venous connection with totally occluded left pulmonary veins presented at our center for fenestrated total cavo-pulmonary connection with an extra cardiac conduit at the age of 3 years. Eleven months after the Fontan completion, she developed protein-losing enteropathy (PLE). Spontaneously closed fenestration was thought to be the cause of the PLE, and she underwent revision of fenestration at the age of 5 years. After the operation, PLE did not improve, and newly developed hypoxemia impaired her systemic ventricular function, leading to the initiation of veno-arterial extracorporeal membrane oxygenation (ECMO) with the Endumo(®) system 18 days after the operation to treat her hemodynamic instability. Although the ECMO circuit was changed three times during the first 8 days, the fourth circuit could be used for 74 days without hemolysis and serum leakage, until the patient unfortunately died 82 days after the operation due to multi-organ failure.


Assuntos
Oxigenação por Membrana Extracorpórea/instrumentação , Cardiopatias Congênitas/cirurgia , Complicações Pós-Operatórias/terapia , Pré-Escolar , Evolução Fatal , Feminino , Cardiopatias Congênitas/complicações , Humanos , Enteropatias Perdedoras de Proteínas/complicações
10.
J Artif Organs ; 16(3): 267-72, 2013 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-23719779

RESUMO

We investigated early clinical outcomes of a new extracorporeal life support (ECLS) system (Endumo 2000, Heiwa Bussan, Tokyo, Japan), which consists of a ROTAFLOW centrifugal pump, a BIOCUBE oxygenator with plasma-leakage-tight polymer fibers, and a biocompatible coating (T-NCVC coating), in pediatric patients <1 year old. From 2008 to 2011, 31 patients required ECLS. Except for 1 patient who was instituted with a transitional ECLS device, a conventional ECLS system (pediatric Emersave, TERUMO, Saitama, Japan) was initiated in 14 patients before December 2009 (6 boys, 63.4 ± 87.1 days old, 3.1 ± 1.0 kg), and the Endumo 2000 was initiated in 16 patients after December 2009 (8 boys, 43.9 ± 78.5 days old, 3.2 ± 0.7 kg). Primary reasons for the institution of ECLS were intraoperative low output syndrome in 11 patients, post-cardiotomy cardiopulmonary collapse in 9 patients, and other reasons in 10 patients. The median support period was 21.7 ± 20.7 days and the total number of circuit exchanges was 83. The median first circuit durability was significantly longer in the Endumo group [8.0 days (range 5.9-13.2) vs. 4.4 days (1.9-8.3)] (p = 0.020). Significant cranial hemorrhage occurred in only 1 patient, who received the Emersave system. The success rate for weaning from ECLS was 14.3% in the Emersave group and 56.3% in the Endumo group. Univariate analysis showed that usage of the Endumo 2000 was a predictor for successful weaning from the ECLS (p = 0.017) as well as survival at discharge (p = 0.032). The Endumo 2000 system provided safe and effective cardiopulmonary support without complications.


Assuntos
Oxigenação por Membrana Extracorpórea/instrumentação , Sistemas de Manutenção da Vida/instrumentação , Oxigenação por Membrana Extracorpórea/efeitos adversos , Feminino , Humanos , Lactente , Recém-Nascido , Masculino , Estudos Retrospectivos , Resultado do Tratamento
11.
J Plant Res ; 125(4): 555-68, 2012 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-22080252

RESUMO

To gain insight into the cellular functions of the mid1-complementing activity (MCA) family proteins, encoding putative Ca²âº-permeable mechanosensitive channels, we isolated two MCA homologs of tobacco (Nicotiana tabacum) BY-2 cells, named NtMCA1 and NtMCA2. NtMCA1 and NtMCA2 partially complemented the lethality and Ca²âº uptake defects of yeast mutants lacking mechanosensitive Ca²âº channel components. Furthermore, in yeast cells overexpressing NtMCA1 and NtMCA2, the hypo-osmotic shock-induced Ca²âº influx was enhanced. Overexpression of NtMCA1 or NtMCA2 in BY-2 cells enhanced Ca²âº uptake, and significantly alleviated growth inhibition under Ca²âº limitation. NtMCA1-overexpressing BY-2 cells showed higher sensitivity to hypo-osmotic shock than control cells, and induced the expression of the touch-inducible gene, NtERF4. We found that both NtMCA1-GFP and NtMCA2-GFP were localized at the plasma membrane and its interface with the cell wall, Hechtian strands, and at the cell plate and perinuclear vesicles of dividing cells. NtMCA2 transcript levels fluctuated during the cell cycle and were highest at the G1 phase. These results suggest that NtMCA1 and NtMCA2 play roles in Ca²âº-dependent cell proliferation and mechanical stress-induced gene expression in BY-2 cells, by regulating the Ca²âº influx through the plasma membrane.


Assuntos
Canais de Cálcio/genética , Canais de Cálcio/metabolismo , Regulação da Expressão Gênica de Plantas/genética , Mecanotransdução Celular/genética , Nicotiana/citologia , Nicotiana/genética , Transporte Biológico/genética , Cálcio/metabolismo , Membrana Celular/metabolismo , Permeabilidade da Membrana Celular/genética , Proliferação de Células , Células Cultivadas , Pressão Osmótica , Plantas Geneticamente Modificadas , Estresse Fisiológico/genética , Nicotiana/metabolismo
12.
Plant J ; 63(1): 141-54, 2010 Jul 01.
Artigo em Inglês | MEDLINE | ID: mdl-20409002

RESUMO

In legumes, Ca(2+)/calmodulin-dependent protein kinase (CCaMK) is a component of the common symbiosis genes that are required for both root nodule (RN) and arbuscular mycorrhiza (AM) symbioses and is thought to be a decoder of Ca(2+) spiking, one of the earliest cellular responses to microbial signals. A gain-of-function mutation of CCaMK has been shown to induce spontaneous nodulation without rhizobia, but the significance of CCaMK activation in bacterial and/or fungal infection processes is not fully understood. Here we show that a gain-of-function CCaMK(T265D) suppresses loss-of-function mutations of common symbiosis genes required for the generation of Ca(2+) spiking, not only for nodule organogenesis but also for successful infection of rhizobia and AM fungi, demonstrating that the common symbiosis genes upstream of Ca(2+) spiking are required solely to activate CCaMK. In RN symbiosis, however, CCaMK(T265D) induced nodule organogenesis, but not rhizobial infection, on Nod factor receptor (NFRs) mutants. We propose a model of symbiotic signaling in host legume plants, in which CCaMK plays a key role in the coordinated induction of infection thread formation and nodule organogenesis.


Assuntos
Proteínas Quinases Dependentes de Cálcio-Calmodulina/metabolismo , Lotus/enzimologia , Proteínas de Plantas/metabolismo , Nodulação/genética , Simbiose/genética , Bactérias/crescimento & desenvolvimento , Cálcio/metabolismo , Proteínas Quinases Dependentes de Cálcio-Calmodulina/genética , Regulação da Expressão Gênica de Plantas , Teste de Complementação Genética , Lotus/genética , Lotus/microbiologia , Mutação , Micorrizas/crescimento & desenvolvimento , Proteínas de Plantas/genética , Transformação Genética
13.
Plant Physiol ; 153(2): 678-92, 2010 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-20357140

RESUMO

Although cytosolic free Ca(2+) mobilization induced by microbe/pathogen-associated molecular patterns is postulated to play a pivotal role in innate immunity in plants, the molecular links between Ca(2+) and downstream defense responses still remain largely unknown. Calcineurin B-like proteins (CBLs) act as Ca(2+) sensors to activate specific protein kinases, CBL-interacting protein kinases (CIPKs). We here identified two CIPKs, OsCIPK14 and OsCIPK15, rapidly induced by microbe-associated molecular patterns, including chitooligosaccharides and xylanase (Trichoderma viride/ethylene-inducing xylanase [TvX/EIX]), in rice (Oryza sativa). Although they are located on different chromosomes, they have over 95% nucleotide sequence identity, including the surrounding genomic region, suggesting that they are duplicated genes. OsCIPK14/15 interacted with several OsCBLs through the FISL/NAF motif in yeast cells and showed the strongest interaction with OsCBL4. The recombinant OsCIPK14/15 proteins showed Mn(2+)-dependent protein kinase activity, which was enhanced both by deletion of their FISL/NAF motifs and by combination with OsCBL4. OsCIPK14/15-RNAi transgenic cell lines showed reduced sensitivity to TvX/EIX for the induction of a wide range of defense responses, including hypersensitive cell death, mitochondrial dysfunction, phytoalexin biosynthesis, and pathogenesis-related gene expression. On the other hand, TvX/EIX-induced cell death was enhanced in OsCIPK15-overexpressing lines. Our results suggest that OsCIPK14/15 play a crucial role in the microbe-associated molecular pattern-induced defense signaling pathway in rice cultured cells.


Assuntos
Morte Celular , Oryza/enzimologia , Proteínas de Plantas/metabolismo , Proteínas Quinases/metabolismo , Sesquiterpenos/metabolismo , Células Cultivadas , DNA Complementar/genética , Regulação da Expressão Gênica de Plantas , Dados de Sequência Molecular , Oryza/genética , Oryza/imunologia , Proteínas de Plantas/genética , Plantas Geneticamente Modificadas/enzimologia , Plantas Geneticamente Modificadas/genética , Plantas Geneticamente Modificadas/imunologia , Proteínas Quinases/genética , RNA de Plantas/genética , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismo , Transdução de Sinais , Técnicas do Sistema de Duplo-Híbrido , Fitoalexinas
14.
J Plant Res ; 124(3): 415-24, 2011 May.
Artigo em Inglês | MEDLINE | ID: mdl-21063744

RESUMO

Microbe/pathogen-associated molecular patterns (MAMPs/PAMPs) often induce rises in cytosolic free Ca(2+) concentration ([Ca(2+)](cyt)) and protein phosphorylation. Though they are postulated to play pivotal roles in plant innate immunity, their molecular links and the regulatory mechanisms remain largely unknown. To investigate the regulatory mechanisms for MAMP-induced Ca(2+) mobilization, we have established a transgenic rice (Oryza sativa) cell line stably expressing apoaequorin, and characterized the interrelationship among MAMP-induced changes in [Ca(2+)](cyt), production of reactive oxygen species (ROS) and protein phosphorylation. Oligosaccharide and sphingolipid MAMPs induced Ca(2+) transients mainly due to plasma membrane Ca(2+) influx, which were dramatically suppressed by a protein phosphatase inhibitor, calyculin A (CA). Hydrogen peroxide and hypo-osmotic shock triggered similar [Ca(2+)](cyt) elevations, which were not affected by CA. MAMP-induced protein phosphorylation, which is promoted by CA, has been shown to be required for ROS production and MAPK activation, while it negatively regulates MAMPs-induced Ca(2+) mobilization and may play a crucial role in temporal regulation of [Ca(2+)](cyt) signature.


Assuntos
Cálcio/metabolismo , Proteínas Quinases Ativadas por Mitógeno/antagonistas & inibidores , Oligossacarídeos/metabolismo , Oryza/metabolismo , Oxazóis/farmacologia , Equorina/genética , Equorina/metabolismo , Apoproteínas/genética , Apoproteínas/metabolismo , Linhagem Celular , Interações Hospedeiro-Patógeno , Imunidade Inata , Toxinas Marinhas , Proteínas Quinases Ativadas por Mitógeno/metabolismo , Oryza/imunologia , Oryza/microbiologia , Fosforilação , Plantas Geneticamente Modificadas/imunologia , Plantas Geneticamente Modificadas/metabolismo , Plantas Geneticamente Modificadas/microbiologia , Espécies Reativas de Oxigênio/metabolismo , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismo
15.
J Electron Microsc (Tokyo) ; 59 Suppl 1: S189-93, 2010 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-20554754

RESUMO

A self-consistent simulation of secondary electron (SE) emission and charging of a SiO(2) layer with the thickness of several tens of nanometres on Si is incorporated into a trajectory simulation of emitted SEs above the surface, the centre area of which is charged by electron beams (EBs) at the energy range from 300 to 2000 eV. In order to study the influence of the charging of an insulating layer on defect inspection, a pseudo-image is reconstructed from net SE yields calculated at each point of the SiO(2) surface locally applied the positive voltage. The image contrast between charged and uncharged areas is compared with the observation of thermally oxidized layer with the thickness of 24-106 nm on a Si wafer. The image contrast is very sensitive to the thickness of the SiO(2) layer, which is verified by both observed and calculated images. The calculated changes of the images with the layer thickness and the primary electron energy reproduce the experimental observations fairly well. This confirms a highly sensitive detection mechanism for tiny defects in insulating patterns on a metal hard mask for an EB defect inspection equipment.

16.
Nat Protoc ; 14(4): 1293-1321, 2019 04.
Artigo em Inglês | MEDLINE | ID: mdl-30903107

RESUMO

Temperature is one of the most important of the physiological parameters that determine the biological status of living organisms. However, intracellular temperature was not imaged at the single-cell level until recently because of the lack of a molecular thermometer that can be applied to living cells. We have recently developed a method for imaging intracellular temperature using a cationic linear fluorescent polymeric thermometer (FPT) and fluorescence lifetime imaging microscopy (FLIM). The cationic linear FPT exhibits cell permeability in various mammalian cell lines and yeast cells, entering live cells within 10 min of incubation. Intracellular thermometry using the cationic linear FPT and FLIM can be used to image temperature with high temperature resolution (0.3-1.29 °C within a temperature range of 25-35 °C). The diffuse intracellular localization of the cationic linear FPT allows a high spatial resolution (i.e., the light microscope's diffraction limit, 200 nm), enabling the detection of temperature distributions at the subcellular level. This protocol, including the construction of a calibration curve and intracellular temperature imaging, requires ~14 h. Experience in handling cultured mammalian cells and use of a confocal laser-scanning microscope (CLSM) is required.


Assuntos
Processamento de Imagem Assistida por Computador/estatística & dados numéricos , Microscopia de Fluorescência/estatística & dados numéricos , Imagem Óptica/estatística & dados numéricos , Saccharomyces cerevisiae/ultraestrutura , Imagem com Lapso de Tempo/estatística & dados numéricos , Animais , Células COS , Linhagem Celular , Chlorocebus aethiops , Células HEK293 , Células HeLa , Humanos , Camundongos , Microscopia de Fluorescência/métodos , Células NIH 3T3 , Imagem Óptica/métodos , Linfócitos T/ultraestrutura , Temperatura , Termômetros , Imagem com Lapso de Tempo/métodos
17.
Polymers (Basel) ; 11(8)2019 Aug 04.
Artigo em Inglês | MEDLINE | ID: mdl-31382693

RESUMO

Cationic nanogels of N-isopropylacrylamide (NIPAM), including NIPAM-based cationic fluorescent nanogel thermometers, were synthesized with a cationic radical initiator previously developed in our laboratory. These cationic nanogels were characterized by transmission electron microscopy (TEM), dynamic light scattering (DLS), zeta potential measurements and fluorescence spectroscopy, as summarized in the temperature-dependent fluorescence response based on the structural change in polyNIPAM units in aqueous solutions. Cellular experiments using HeLa (human epithelial carcinoma) cells demonstrated that NIPAM-based cationic fluorescent nanogel thermometers can spontaneously enter the cells under mild conditions (at 25 °C for 20 min) and can show significant fluorescence enhancement without cytotoxicity with increasing culture medium temperature. The combination of the ability to enter cells and non-cytotoxicity is the most important advantage of cationic fluorescent nanogel thermometers compared with other types of fluorescent polymeric thermometers, i.e., anionic nanogel thermometers and cationic/anionic linear polymeric thermometers.

18.
J Org Chem ; 73(19): 7811-3, 2008 Oct 03.
Artigo em Inglês | MEDLINE | ID: mdl-18754642

RESUMO

The gold(I) complex (MeAuPPh3) was found to be a highly effective catalyst for the hydrative cyclization of 1,6-diynes to form the corresponding 3-methyl hex-2-enone derivatives with good to excellent yield. The proposed mechanism is described.


Assuntos
Cicloexenos/síntese química , Di-Inos/química , Catálise , Ouro/química , Cetonas
19.
Ther Apher Dial ; 21(2): 166-172, 2017 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-28296183

RESUMO

It currently remains unclear whether stroke volume variation (SVV) before hemodialysis (HD) is an independent predictor of decreased blood pressure (BP) during HD. Fifty-two patients were divided into two groups (Decreased BP during HD group: N = 10, Non-decreased BP group: N = 42). Fractional shortening was lower, and mean arterial pressure (MAP) and SVV were higher in the Decreased BP during HD group. A multiple logistic regression analysis identified low fractional shortening, high MAP, and high SVV as independent predictors of decreased BP during HD. The areas under the ROC curves were as follows: 0.849 for MAP, 0.712 for SVV, and 0.893 for MAP and SVV. Optimal threshold values were 93.0 mm Hg for MAP and 17.3 % for SVV. A multivariate regression analysis identified anemia and a longer dialysis vintage as independently related factors for higher SVV. Our results suggest that high SVV is an independent predictor for decreased BP during HD.


Assuntos
Hipotensão/diagnóstico , Diálise Renal , Volume Sistólico/fisiologia , Idoso , Pressão Sanguínea/fisiologia , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Curva ROC
20.
ASAIO J ; 52(1): 110-4, 2006.
Artigo em Inglês | MEDLINE | ID: mdl-16436900

RESUMO

Quick setup is mandatory for cardiopulmonary resuscitation using an extracorporeal membrane oxygenation (ECMO) assist device. Our conventional ECMO circuit for pediatric patients consists of a centrifugal pump (CX-HP) and membrane oxygenator (CX10H). Because of the large priming volume (260 ml), the circuit had to be primed with donor blood and required 30 minutes for setup. We started to use a low-prime ECMO with small centrifugal pump (HPM-15) and membrane oxygenator (MENOX Alpha Cube) for induction of ECMO beginning in 2000. The priming volume of this low-prime circuit is only 99 ml. The circuit can be primed without donor blood, even in the small patient, and requires only 10 minutes to set up. We review our experiences with cardiopulmonary resuscitation for sudden cardiopulmonary collapse in pediatric patients, including postcardiotomy patients. From 1997 to 2000, 23 patients underwent ECMO support with a conventional circuit (group A). From 2000 to 2004, we used low-prime circuit for induction of ECMO in 12 patients (group B). After the induction of ECMO with low-prime circuit, ECMO was converted to conventional heparin-bonded circuit for the longer support. The results suggested that the quick induction of ECMO with low-prime circuit has significant advantages in cardiopulmonary support in pediatric patients.


Assuntos
Oxigenação por Membrana Extracorpórea/instrumentação , Oxigenação por Membrana Extracorpórea/métodos , Reanimação Cardiopulmonar/estatística & dados numéricos , Criança , Pré-Escolar , Humanos , Lactente , Recém-Nascido , Japão/epidemiologia , Oxigenadores de Membrana , Estudos Retrospectivos , Resultado do Tratamento
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