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1.
Front Rehabil Sci ; 3: 1019089, 2022.
Artículo en Inglés | MEDLINE | ID: mdl-36569638

RESUMEN

Background: As a type of welfare technology, care robotics is now widely seen as a potential aide to rehabilitation, increasing independence and enhancing the wellbeing of people with disabilities and older adults. Research into and development of care robots have both been vigorously promoted in North America, Europe and Asia, and the competition for technological advancement in robotics is becoming fierce. AI ethics and policy guidelines are being established. However, there are still differences in attitudes and perceptions, as well as national policies regarding this type of welfare technology. Moreover, despite the anticipated usefulness, it is believed that progress has been slow in the diffusion of care robots. Purpose: In order to explore how public discourses support technological innovation, such as care robots, while preparing society for potential risks and impact, we sought to ascertain whether public discourse on care robots varies from region to region. For example, what are the hopes and promises associated with care robots and what are the concerns? Methods: To address these questions, this article explored how care robots have been portrayed in five major broadsheet newspapers in five jurisdictions in Asia and Europe (France, Great Britain, Hong Kong SAR, Ireland and Japan). We obtained 545 articles for the period between January 2001 and September 2020, more than half of which originated in Japan. A thematic analysis was conducted of these articles written in four languages (Chinese, English, French and Japanese). Results: Positive and negative narratives were teased out, alongside other key prominent themes identified, such as Japan as the land of robots, the pandemic, and the impact of robots on the economy. As the number of robot-related articles grew from the year 2012 onwards, narratives became more nuanced in European newspapers, but not in Asian ones. Furthermore, recent articles began to address the social and relational impact of care robots, while providing concrete examples of improvements in the quality of life for users. Further careful examination will be necessary in the future in order to establish the impact of robotics use in rehabilitation for people with disabilities, older adults, their carers and society at large.

2.
Oral Dis ; 18(5): 506-12, 2012 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-22309644

RESUMEN

OBJECTIVE: This investigation was a basal study that used a mouse model of xerostomia to identify protein biomarkers of xerostomia in saliva. We identified genes expressed differently in parotid glands from non-obese diabetic mice with diabetes and those from control mice; subsequently, we investigated expression of the proteins encoded by these genes in parotid glands and saliva. MATERIALS AND METHODS: DNA microarray and real-time PCR analyses were performed to detect differences between NOD/ShiJcl and C57BL/6JJcl (control) female mice in gene expression from parotid glands or parotid acinar cells. Subsequently, protein expression was assessed using immunoblotting and immunohistochemistry. Similarly, enzyme activity in saliva was assessed using zymography. RESULTS: Based on gene expression analyses, Chia expression was higher in diabetic mice than non-diabetic mice and control mice; similarly, expression of chitinase, the protein encoded by Chia, was higher in diabetic mice. Saliva from NOD/ShiJcl mice had more chitinase than saliva from control mice. CONCLUSIONS: Chitinase was highly expressed in parotid acinar cells from diabetic mice compared with non-diabetic and control mice. Increased chitinase expression and enzyme activity may characterize the autoimmune diabetes in mice; however, further investigation is required to assess its use as a biomarker of xerostomia in humans.


Asunto(s)
Quitinasas/genética , Diabetes Mellitus Tipo 1/enzimología , Glándula Parótida/enzimología , Saliva/enzimología , Xerostomía/enzimología , Células Acinares/enzimología , Animales , Quitinasas/biosíntesis , Diabetes Mellitus Tipo 1/genética , Modelos Animales de Enfermedad , Femenino , Expresión Génica , Ratones , Ratones Endogámicos C57BL , Ratones Endogámicos NOD , Análisis de Secuencia por Matrices de Oligonucleótidos , Glándula Parótida/citología , Proteínas y Péptidos Salivales/biosíntesis
3.
Ann Oncol ; 22(9): 2113-2120, 2011 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-21307157

RESUMEN

BACKGROUND: To identify factors influencing place of death among home palliative care patients with advanced cancer, focusing on the timing of referrals from hospital to home care settings. METHODS: A cross-sectional nationwide questionnaire survey was conducted on home palliative care patients at 1000 randomly selected home care agencies in Japan. A total of 568 responses were analyzed (effective response rate, 69%). RESULTS: Multivariate logistic regression analysis revealed that (i) predischarge health care supports in hospital (e.g. early referral 8 days or more before discharge; clear explanation by hospital staffs to patients and families regarding discharge to live and die at home) and (ii) postdischarge health care supports after transferring home care (e.g. signing a 24-h support insurance contract of network between primary physician and nurse as a home palliative care team; primary nurse consultation with primary physician >3 times during the first week after discharge) have an effect on place of death among home palliative care patients. CONCLUSION: An early and carefully coordinated referral support system for smooth discharge by hospital staffs as well as intensive and highly qualified support just after discharge by the home care team would help to increase the number of patients who could die at home.


Asunto(s)
Actitud Frente a la Muerte , Servicios de Atención de Salud a Domicilio/organización & administración , Neoplasias/terapia , Cuidados Paliativos/organización & administración , Anciano , Estudios Transversales , Relaciones Familiares , Femenino , Agencias de Atención a Domicilio/organización & administración , Agencias de Atención a Domicilio/estadística & datos numéricos , Servicios de Atención de Salud a Domicilio/estadística & datos numéricos , Humanos , Japón , Modelos Logísticos , Masculino , Neoplasias/psicología , Cuidados Paliativos/métodos , Cuidados Paliativos/estadística & datos numéricos , Derivación y Consulta , Encuestas y Cuestionarios , Enfermo Terminal
4.
Clin Exp Immunol ; 134(2): 279-84, 2003 Nov.
Artículo en Inglés | MEDLINE | ID: mdl-14616788

RESUMEN

Chemotherapy causes neutropenia and an increased susceptibility to infection. Recent reports indicate that mannan-binding lectin (MBL) insufficiency is associated with an increased duration of febrile neutropenia and incidence of serious infections following chemotherapy for haematological malignancies. We aimed to confirm or refute this finding and to extend the investigation to the plasma ficolins, P35 (L-ficolin) and the Hakata antigen (H-ficolin). MBL, L-ficolin and H-ficolin were measured in 128 patients with haematological malignancies treated by chemotherapy alone or combined with bone marrow transplantation. Protein concentrations were related to clinical data retrieved from medical records. MBL concentrations were elevated compared with healthy controls in patients who received chemotherapy, while L-ficolin concentrations were decreased and H-ficolin levels were unchanged. There was no correlation between MBL, L-ficolin or H-ficolin concentration and febrile neutropenia expressed as the proportion of neutropenic periods in which patients experienced fever, and there was no relation between abnormally low (deficiency) levels of MBL, L-ficolin or H-ficolin and febrile neutropenia so expressed. Patients with MBL < or =0.1 microg/ml had significantly more major infections than no infections within the follow-up period (P<0.05), but overall most patients had signs or symptoms of minor infections irrespective of MBL concentration. Neither L-ficolin nor H-ficolin deficiencies were associated with infections individually, in combination or in combination with MBL deficiency. MBL, L-ficolin and H-ficolin, independently or in combination, did not have a major influence on susceptibility to infection in these patients rendered neutropenic by chemotherapy. These results cast doubt on the potential value of MBL replacement therapy in this clinical context.


Asunto(s)
Antineoplásicos/efectos adversos , Proteínas Portadoras/sangre , Lectinas , Lectina de Unión a Manosa/sangre , Infecciones Oportunistas/sangre , Adolescente , Adulto , Anciano , Anciano de 80 o más Años , Antineoplásicos/uso terapéutico , Susceptibilidad a Enfermedades , Femenino , Neoplasias Hematológicas/tratamiento farmacológico , Neoplasias Hematológicas/inmunología , Humanos , Huésped Inmunocomprometido/inmunología , Masculino , Persona de Mediana Edad , Neutropenia/inducido químicamente , Neutropenia/complicaciones , Neutropenia/inmunología , Infecciones Oportunistas/complicaciones , Índice de Severidad de la Enfermedad , Ficolinas
5.
Biol Pharm Bull ; 24(6): 650-5, 2001 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-11411553

RESUMEN

"The extract of shikon" (SK) and shikonin play important roles in the development of granulomatous tissue formation. To reveal the augmenting effect of SK or shikonin on vascular endothelial growth factor (VEGF) production and neovascularization, we investigated murine granulomatous tissue induced by SK and shikonin, comparing them to pouches in which trehalose 6,6'-dimycolate (TDM) was injected. The development of granulomatous tissue formation was evaluated by the wet weight of pouch walls. At day 5 and 7 after SK and shikonin injection, prominent granulomatous tissue formation was detected. Histological observations on the development of granulomatous tissue showed that the pouch was formed in the submuscular connective tissue and necrotic tissue directly facing the cavity and granulomatous tissue developed in the connective tissue. At day 1, VEGF-positive neutrophils accumulated in the pouch wall. Granulomatous tissue formation and neovascularization by injection of SK or shikonin was not more prominent than TDM. However, the present results indicate that SK and shikonin induce neovascularization in granulomatous tissue.


Asunto(s)
Granuloma/inducido químicamente , Naftoquinonas/farmacología , Animales , Complejo CD3/biosíntesis , Factores de Crecimiento Endotelial/biosíntesis , Citometría de Flujo , Granuloma/inmunología , Linfocinas/biosíntesis , Antígeno de Macrófago-1/biosíntesis , Masculino , Ratones , Ratones Endogámicos ICR , Naftoquinonas/toxicidad , Factor A de Crecimiento Endotelial Vascular , Factores de Crecimiento Endotelial Vascular
6.
J Inorg Biochem ; 83(4): 247-53, 2001 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-11293544

RESUMEN

The characteristic features of Fe-type nitrile hydratase (NHase) from Rhodococcus sp. N-771 are described. Through the biochemical analyses, we have found that nitric oxide (NO) regulates the photoreactivity of this enzyme by association with the non-heme iron center and photoinduced dissociation from it. The regulation is realized by a unique structure of the catalytic non-heme iron center composed of post-translationally modified cysteine-sulfinic (Cys-SO2H) and -sulfenic acids (Cys-SOH). To understand the biogenic mechanism and the functional role of these modifications, we constructed an over-expression system of whole NHase and individual subunits in Escherichia coli. The results of the studies on several recombinant NHases have shown that the Cys-SO2H oxidation of alphaC112 is indispensable for the catalytic activity of Fe-type NHase.


Asunto(s)
Hidroliasas/química , Hidroliasas/metabolismo , Hierro/metabolismo , Rhodococcus/enzimología , Sitios de Unión , Hidroliasas/genética , Modelos Moleculares , Conformación Proteica , Proteínas Recombinantes/química , Proteínas Recombinantes/metabolismo , Rhodococcus/genética
7.
Clin Diagn Lab Immunol ; 8(2): 454-9, 2001 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-11238239

RESUMEN

Although a serum thermolabile beta-2 macroglycoprotein (TMG) may play a role in host defense as a lectin, little is known of its related physiological functions, mainly due to a lack of appropriate methods for tracing the functions of TMG. We identified a polysaccharide from Aerococcus viridans, PSA, which reacts with TMG, and based on this finding, we developed an enzyme-linked immunosorbent assay to trace the functions of TMG. Using ethanol precipitation and DEAE-Sepharose and Sephacryl S-400 column chromatographies, we isolated PSA from cultured medium of A. viridans, and it exhibited specific binding against TMG in blood samples. In sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), the isolated PSA showed ladder bands that implied the existence of repeating units composed of D-glucose, N-acetyl-D-glucosamine, D-mannose, and D-xylose, as confirmed by gas chromatography-mass spectrometry. SDS-PAGE and immunochemical analysis, using rabbit anti-TMG antibody, showed that PSA specifically binds solely to intact serum TMG but not to TMG heated at 56 degrees C for 30 min, a condition under which antigenicity is lost. TMG in serum samples bound to PSA in a dose-dependent manner, and this binding was clearly suppressed by addition of PSA. These observations indicate that PSA is a useful adsorbent to TMG and can be used to develop appropriate methods for tracing the functions of TMG.


Asunto(s)
Ensayo de Inmunoadsorción Enzimática/métodos , Glicoproteínas/análisis , Polisacáridos Bacterianos/inmunología , Streptococcaceae/inmunología , Anticuerpos Antibacterianos/inmunología , Autoanticuerpos/sangre , Autoanticuerpos/inmunología , Glicoproteínas/inmunología , Glicoproteínas/metabolismo , Humanos , Immunoblotting , Lectinas , Lupus Eritematoso Sistémico/inmunología , Polisacáridos Bacterianos/aislamiento & purificación , Polisacáridos Bacterianos/metabolismo , Pruebas de Precipitina , Unión Proteica/inmunología
8.
Percept Mot Skills ; 91(1): 69-78, 2000 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-11011873

RESUMEN

The subjects performed an orienting task involving 3 conditions, followed by unexpected tests which included free recall, name-matching and name-selec tion. Conditions were designed to force self-generated elaboration, self-choice elaboration, and experimenter-provided elaboration. In the self-generated elaboration condition, subjects were presented target sentences, e.g., Nobunaga ODA burned down ENRYAKUJI Temple, and asked to answer an elaborative interrogation, e.g., Why did Nobunaga ODA burn down ENRYAKUJI Temple? about each sentence. In the self-choice elaboration condition, subjects selected one of the alternative answers to an elaborative interrogation about each sentence. In the experimenter provided elaboration condition, subjects were presented an answer which they rated for congruity as the correct answer to the elaborative interrogation. In the free recall test, self generated elaboration led to better performance than the other two conditions for which no difference was observed. However, in the name-matching and name-selection tests, scores were better for self choice elaboration and self-generated elaboration than for experimenter-provided elaboration. These results were interpreted as demonstrating that self choice elaboration, in addition to self-generated elaboration, led to effective encoding in memory.


Asunto(s)
Aprendizaje por Asociación , Conducta de Elección , Memoria , Conducta Verbal , Adolescente , Femenino , Humanos , Lingüística , Masculino , Recuerdo Mental , Semántica
9.
J Inorg Biochem ; 80(3-4): 283-8, 2000 Jul 01.
Artículo en Inglés | MEDLINE | ID: mdl-11001100

RESUMEN

Arginine 56 in the beta subunit (betaArg56) of the iron-containing nitrile hydratase (NHase), one of the strongly conserved residues within the NHase family, is known to form hydrogen bonds to the sulfinyl (-SO2H) and sulfenyl (-SOH) groups of the post-translationally modified cysteine residues in the catalytic center. BetaArg56 was substituted by tyrosine, glutamate or lysine, respectively, and the respective mutant enzymes generated by reconstitution were characterized. The betaR56K mutant complex exhibited about 1% of the enzymatic activity of native NHase, while the others were totally inactive. The kinetic analysis of the betaR56K mutant complex exhibited a drastic decrease in turnover number and decreases in kinetic constants for substrate and inhibitors as compared to the native NHase. Changes in UV-visible absorption and light-induced Fourier transform infrared difference spectra suggest that betaArg56 is involved in the positioning of the -SO2H and -SOH groups of the modified Cys residues in the catalytic center so as to fine tune the electronic state of the iron center suitable for catalysis. Thus, betaArg56 is essential for catalysis.


Asunto(s)
Arginina/química , Escherichia coli/enzimología , Hidroliasas/química , Enlace de Hidrógeno , Arginina/genética , Sitios de Unión , Clonación Molecular , Escherichia coli/genética , Hidroliasas/genética , Hidroliasas/metabolismo , Cinética , Estructura Molecular , Mutagénesis Sitio-Dirigida , Proteínas Recombinantes/química , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismo , Análisis Espectral
10.
Brain Res Bull ; 52(4): 291-6, 2000 Jul 01.
Artículo en Inglés | MEDLINE | ID: mdl-10856827

RESUMEN

An overactive brain renin-angiotensin system is one of the factors contributing to the pathogenesis of hypertension in spontaneously hypertensive rats (SHR). We examined brain sites where enhanced activity of an angiotensin system is responsible for the pathogenesis of hypertension in SHR. The angiotensin receptor antagonist, losartan was injected into tissues around rostral parts of the third ventricle in conscious rats. Losartan (0.22 nmol) injected into the anterior hypothalamic area, anterior (AHA) produced a depressor response in SHR but not in Wistar Kyoto rats (WKY). Angiotensin II (0.091-0.91 pmol) injected into the AHA produced a pressor response in both WKY and SHR, and the pressor response to angiotensin II was greater in SHR than that of WKY. Carbachol (3 pmol) injected into the AHA also produced a pressor response in WKY and SHR, and the pressor response to carbachol was almost the same in both strains of rats. Release of angiotensin peptides in the AHA was greater in SHR than that of WKY. These findings suggest that an angiotensin system in the AHA is enhanced and this enhancement of angiotensin system is involved in the maintenance of hypertension in SHR. Both increased pressor reactivity to angiotensin II and increased release of angiotensin peptides in the AHA appear to be related to this enhancement of angiotensin system in SHR.


Asunto(s)
Angiotensinas/metabolismo , Núcleo Hipotalámico Anterior/metabolismo , Hipertensión/metabolismo , Angiotensina II/administración & dosificación , Animales , Núcleo Hipotalámico Anterior/efectos de los fármacos , Antihipertensivos/administración & dosificación , Presión Sanguínea/efectos de los fármacos , Carbacol/administración & dosificación , Hipertensión/etiología , Losartán/administración & dosificación , Masculino , Microinyecciones , Perfusión , Ratas , Ratas Endogámicas SHR , Ratas Endogámicas WKY
11.
Hypertens Res ; 23(2): 109-18, 2000 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-10770257

RESUMEN

It has been established that deoxycorticosterone acetate (DOCA)-salt hypertensive rats have an overactive brain angiotensin-system. The purpose of the present study was to identify the brain sites showing enhanced angiotensin-system activity responsible for the pathogenesis of hypertension in DOCA-salt hypertensive rats. The angiotensin receptor antagonist, losartan, was injected into brain ventricles or into tissues around the rostral parts of the third ventricle in conscious DOCA-salt hypertensive rats. Losartan (1 microg) injection into the lateral ventricle or into the rostral parts of the third ventricle produced a depressor response, whereas the agent did not affect blood pressure when injected into the caudal parts of the third ventricle or into the fourth ventricle. Losartan (0.1 microg) injection into the anterior hypothalamic preoptic area, anterior (AHA) produced a depressor response. Angiotensin II (0.1-1 ng) injection into the AHA produced a pressor response in sham-operated and DOCA-salt hypertensive rats, and the pressor response to angiotensin II (1 ng) was greater in DOCA-salt hypertensive rats than that in sham-operated rats. Release of angiotensin peptides in the AHA was greater in DOCA-salt hypertensive rats than that in sham-operated rats. These findings suggest that the angiotensin-system in the AHA is enhanced, and that this enhancement is involved in the maintenance of hypertension in DOCA-salt hypertensive rats. Both increased pressor reactivity to angiotensin II and increased release of angiotensin peptides in the AHA appear to be related to this enhancement of the angiotensin-system in DOCA-salt hypertensive rats.


Asunto(s)
Presión Sanguínea/efectos de los fármacos , Hipertensión/fisiopatología , Hipotálamo Anterior/metabolismo , Área Preóptica/metabolismo , Receptores de Angiotensina/fisiología , Angiotensina II/farmacología , Antagonistas de Receptores de Angiotensina , Animales , Antihipertensivos/farmacología , Desoxicorticosterona , Relación Dosis-Respuesta a Droga , Hipertensión/inducido químicamente , Inyecciones Intraventriculares , Losartán/farmacología , Masculino , Ratas , Ratas Wistar , Valores de Referencia , Cloruro de Sodio , Vasoconstrictores/farmacología
12.
J Biochem ; 125(4): 696-704, 1999 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-10101282

RESUMEN

The nitrile hydratase (NHase) from Rhodococcus sp. N-771 is a photoreactive enzyme that is inactivated on nitrosylation of the non-heme iron center and activated on photo-dissociation of nitric oxide (NO). The nitrile hydratase operon consists of six genes encoding NHase regulator 2, NHase regulator 1, amidase, NHase alpha subunit, NHase beta subunit and NHase activator. We overproduced the NHase in Escherichia coli using a T7 expression system. The NHase was functionally expressed in E. coli only when the NHase activator encoded downstream of the beta subunit gene was co-expressed and the transformant was grown at 30 degrees C or less. A ligand cysteine, alphaCys112, of the recombinant NHase was also post-translationally modified to a cysteine-sulfinic acid similar to for the native NHase. Although another modification of alphaCys114 could not be identified because of the instability under acidic conditions, the recombinant NHase could be reversibly inactivated by nitric oxide.


Asunto(s)
Escherichia coli/enzimología , Escherichia coli/genética , Hidroliasas/genética , Hidroliasas/metabolismo , Secuencia de Aminoácidos , Secuencia de Bases , Clonación Molecular , Cisteína/química , Cartilla de ADN/genética , Activación Enzimática , Expresión Génica , Genes Bacterianos , Hidroliasas/química , Ligandos , Datos de Secuencia Molecular , Operón , Pliegue de Proteína , Procesamiento Proteico-Postraduccional , Proteínas Recombinantes/química , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismo , Rhodococcus/enzimología , Rhodococcus/genética , Homología de Secuencia de Aminoácido , Solubilidad , Temperatura
13.
Nat Struct Biol ; 5(5): 347-51, 1998 May.
Artículo en Inglés | MEDLINE | ID: mdl-9586994

RESUMEN

The iron-containing nitrile hydratase (NHase) is a photoreactive enzyme that is inactivated in the dark because of persistent association with NO and activated by photo-dissociation of NO. The crystal structure at 1.7 A resolution and mass spectrometry revealed the structure of the non-heme iron catalytic center in the nitrosylated state. Two Cys residues coordinated to the iron were post-translationally modified to Cys-sulfenic and -sulfinic acids. Together with another oxygen atom of the Ser ligand, these modifications induced a claw setting of oxygen atoms capturing an NO molecule. This unprecedented structure is likely to enable the photo-regulation of NHase and will provide an excellent model for designing photo-controllable chelate complexes and, ultimately, proteins.


Asunto(s)
Proteínas Bacterianas/química , Hidroliasas/química , Proteínas de Hierro no Heme/química , Oxígeno/química , Proteínas Bacterianas/metabolismo , Sitios de Unión , Cristalografía por Rayos X , Hidroliasas/metabolismo , Modelos Biológicos , Modelos Moleculares , Proteínas de Hierro no Heme/metabolismo , Conformación Proteica , Procesamiento Proteico-Postraduccional
14.
Nihon Rinsho Meneki Gakkai Kaishi ; 20(5): 453-6, 1997 Oct.
Artículo en Japonés | MEDLINE | ID: mdl-9391310

RESUMEN

A 25-year-old woman was admitted for general arthralgia in July, 1989. Reactive arthritis with arthralgia after Shigellosis was diagnosed by sex, localization of arthralgia and positive for HLA-B 27. Within 3 weeks after starting diclifenac sodium 75 mg/day, the arthralgia remitted. It has been reported that patients who are positive for HLA-B 27 have a more severe acute or chronic sacroiliitis, and our case may support this report.


Asunto(s)
Artritis Reactiva/etiología , Disentería Bacilar/complicaciones , Adulto , Antiinflamatorios/uso terapéutico , Artralgia/tratamiento farmacológico , Artralgia/etiología , Artritis Reactiva/tratamiento farmacológico , Biomarcadores/sangre , Diclofenaco/uso terapéutico , Femenino , Antígeno HLA-B27/sangre , Humanos , Esteroides
15.
J Biol Chem ; 272(47): 29454-9, 1997 Nov 21.
Artículo en Inglés | MEDLINE | ID: mdl-9368004

RESUMEN

Nitrile hydratase (NHase) from Rhodococcus sp. N-771 is a photoreactive enzyme that is inactivated by nitrosylation of the non-heme iron center and activated by photodissociation of nitric oxide (NO). To obtain structural information on the iron center, we isolated peptide complexes containing the iron center by proteolysis. When the tryptic digest of the alpha subunit isolated from the inactive form was analyzed by reversed-phase high performance liquid chromatography, the absorbance characteristic of the nitrosylated iron center was observed in the peptide fragment, Asn105-Val-Ile-Val-Cys-Ser-Leu-Cys-Ser-Cys-Thr-Ala-Trp-Pro-Ile-Leu - Gly-Leu-Pro-Pro-Thr-Trp-Tyr-Lys128. The peptide contained 0.79 mol of iron/mol of molecule as well as endogenous NO. Subsequently, by digesting the peptide with thermolysin, carboxypeptidase Y, and leucine aminopeptidase M, we found that the minimum peptide segment required for the nitrosylated iron center is the 11 amino acid residues from alphaIle107 to alphaTrp117. Furthermore, by using mass spectrometry, protein sequence, and amino acid composition analyses, we have shown that the 112th Cys residue of the alpha subunit is post-translationally oxidized to a cysteine-sulfinic acid (Cys-SO2H) in the NHase. These results indicate that the NHase from Rhodococcus sp. N-771 has a novel non-heme iron enzyme containing a cysteine-sulfinic acid in the iron center. Possible ligand residues of the iron center are discussed.


Asunto(s)
Cisteína/metabolismo , Hidroliasas/química , Procesamiento Proteico-Postraduccional , Rhodococcus/enzimología , Secuencia de Aminoácidos , Cromatografía Líquida de Alta Presión , Espectroscopía de Resonancia por Spin del Electrón , Hidroliasas/biosíntesis , Hierro/metabolismo , Datos de Secuencia Molecular , Mapeo Peptídico , Fotoquímica , Espectrometría de Masa por Láser de Matriz Asistida de Ionización Desorción , Espectrofotometría Ultravioleta
16.
Nihon Rinsho Meneki Gakkai Kaishi ; 20(1): 8-13, 1997 Feb.
Artículo en Japonés | MEDLINE | ID: mdl-9105160

RESUMEN

The levels of IgG subclasses were determined in patients with active systemic lupus erythematosus (SLE). The levels of all IgG subclasses in these patients were higher than normal controls. However, there were variations in the increase of IgG subclasses, especially IgG 1 or IgG 2. As concerning the relation to the clinical findings, the levels of IgG 1 significantly increased in patients with renal involvements and those of IgG 3 increased in patients with low complements. Thus, it was suggested that some IgG subclass related to some clinical findings.


Asunto(s)
Inmunoglobulina G/análisis , Lupus Eritematoso Sistémico/inmunología , Humanos
17.
Biochemistry ; 35(51): 16777-81, 1996 Dec 24.
Artículo en Inglés | MEDLINE | ID: mdl-8988015

RESUMEN

Nitrile hydratase (NHase) from Rhodococcus sp. N-771, which contains a non-heme iron center in the catalytic site, has been known to be activated by light illumination. Recently, endogenous nitric oxide (NO) was found in this enzyme by FTIR spectroscopy [Noguchi et al. (1995) FEBS Lett. 358, 9-12]. In order to directly detect the bonding between NO and the iron atom and the reaction of NO upon photoactivation, resonance Raman spectra of the NHase were measured with 413 nm excitation at 85 K. Two prominent bands at 592 and 570 cm-1 were observed in the inactive from, and both of them were completely lost upon photoactivation. Upon subsequent introduction of 15NO, the active NHase was converted to the inactive form again and the above two bands were restored with downshifts by 10 and 12 cm-1, respectively. Also, the excitation profiles of these bands in the 350-500 nm region mostly followed the absorption spectrum arising from the iron center. From these isotopic shifts and the excitation profiles, the two Raman bands were assigned to the Fe-NO stretching and bending vibrations that are probably coupled with each other. The results provided solid evidence that NO is bound to the non-heme iron in the inactive NHase and its photodissociation activates the enzyme.


Asunto(s)
Hidroliasas/química , Hidroliasas/metabolismo , Rhodococcus/enzimología , Sitios de Unión , Activación Enzimática/efectos de la radiación , Hidroliasas/efectos de la radiación , Hierro/química , Óxido Nítrico/química , Fotoquímica , Espectroscopía Infrarroja por Transformada de Fourier , Espectrometría Raman
18.
J Biochem ; 119(3): 407-13, 1996 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-8830032

RESUMEN

Nitrile hydratase (NHase) from Rhodococcus sp. N-771 exists in active and inactive forms. The inactive NHase is immediately activated by light irradiation and changes to the active form. To characterize the photoreactive center, the inactive NHase was denatured by 6 M urea, and two kinds of subunits (alpha and beta) were separated and purified by anion-exchange chromatography. In a manner similar to the native NHase, the isolated alpha subunit showed two absorption peaks at 280 and 370 nm, which were diminished by light irradiation. However, irradiation failed to elicit the appearance of absorption peaks at around 400 nm and at 710 nm, which were characteristic of the activated enzyme. The beta subunit seemed not to possess any photoreactive chromophore because its absorption spectrum was not altered by light irradiation. Neither of the subunits showed NHase activity before and after light irradiation, but the inactive NHase was reconstituted by incubating the two subunits together in the dark at 4 degrees C for 1 h. Light irradiation of the beta subunit did not affect subsequent complex formation or NHase activity. However, the irradiated alpha subunit could not assemble with the beta subunit, and no activity was recovered. These results demonstrate that the chromophore(s) responsible for the photoactivation of NHase are entirely located on the alpha subunit, and imply that light irradiation induces conformational change of the alpha subunit.


Asunto(s)
Hidroliasas/química , Sitios de Unión , Cromatografía en Gel , Cromatografía por Intercambio Iónico , Hidroliasas/efectos de la radiación , Fotoquímica , Conformación Proteica , Rhodococcus/enzimología , Espectrofotometría Atómica
19.
Vox Sang ; 69(3): 206-12, 1995.
Artículo en Inglés | MEDLINE | ID: mdl-8578732

RESUMEN

Using an immunodiffusion assay, we tested all of the blood units donated at the Fukuoka Red Cross Blood Center from June 1991 to July 1994 for B19 antigen. Over this 3-year trial period, we detected 16 viremic cases out of approximately 560,000 blood donors. Interestingly, most of the viremic donors (15 out of 16) were detected between February 1992 and January 1993, which coincided with a local erythema infectiosum epidemic in the Fukuoka area (December 1991 to August 1992). In particular, we detected 4 cases of viremia in March 1992, which was the peak of the erythema infectiosum epidemic. The incidence of B19 viremia in this peak period was approximately 1/4,000. The viremic donors ranged in age from 17 to 45 years, and most (11/16) were between 31 and 39 years old. By ELISA, using virus particles purified from viremic donor plasma as antigen, we analyzed the prevalence of B19-specific antibody among blood donors. The antibody-positive rate was approximately 40% in donors 16-30 years old, gradually increased in middle age, and reached a peak of 92% in donors more than 61 years old.


Asunto(s)
Antígenos Virales/sangre , Donantes de Sangre , Eritema Infeccioso/sangre , Tamizaje Masivo/métodos , Parvovirus B19 Humano/aislamiento & purificación , Adolescente , Adulto , Anticuerpos Antivirales/sangre , Especificidad de Anticuerpos , Ensayo de Inmunoadsorción Enzimática , Eritema Infeccioso/epidemiología , Femenino , Humanos , Japón/epidemiología , Masculino , Persona de Mediana Edad , Parvovirus B19 Humano/inmunología , Prevalencia
20.
Int J Vitam Nutr Res ; 65(1): 36-9, 1995.
Artículo en Inglés | MEDLINE | ID: mdl-7657479

RESUMEN

The antiscorbutic activity of 6-bromo-6-deoxy-L-ascorbic acid (I) in guinea pigs was compared to that of ascorbic acid (AsA). The growth test results in guinea pigs did not show any significant difference between the body weight increase in the respective 2 groups given compound (I) and that in the group given AsA. Scorbutic guinea pigs were found to completely recover from the disease after administration of the compound (I). There were no significant differences in the plasma alkaline phosphatase activity and the hydroxyproline content in the tibia between the group given compound (I) and that given AsA. The total vitamin C contents in the various tissues were similar in the 2 groups given compound (I) and the C-deficient group but they were lower than the levels for the group given AsA. It was shown that antiscorbutic activity was characteristic of compound (I) and it was almost equal to the effect of AsA.


Asunto(s)
Ácido Ascórbico/análogos & derivados , Escorbuto/tratamiento farmacológico , Fosfatasa Alcalina/sangre , Animales , Ácido Ascórbico/análisis , Ácido Ascórbico/uso terapéutico , Peso Corporal/efectos de los fármacos , Química Encefálica , Cobayas , Hidroxiprolina/análisis , Riñón/química , Hígado/química , Masculino , Bazo/química , Tibia/química
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