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1.
Nanotechnology ; 28(27): 275702, 2017 Jul 07.
Artículo en Inglés | MEDLINE | ID: mdl-28525395

RESUMEN

The room temperature photoluminescence from ZnO/MgO core/shell nanowires (NWs) grown by a simple two-step vapor transport method was studied for various MgO shell widths (w). Two distinct effects induced by the MgO shell were clearly identified. The first one, related to the ZnO/MgO interface formation, is evidenced by strong enhancements of the zero-phonon and first phonon replica of the excitonic emission, which are accompanied by a total suppression of its second phonon replica. This effect can be explained by the reduction of the band bending within the ZnO NW core that follows the removal of atmospheric adsorbates and associated surface traps during the MgO growth process on one hand, and a reduced exciton-phonon coupling as a result of the mechanical stabilization of the outermost ZnO NW monolayers by the MgO shell on the other hand. The second effect is the gradual increase of the excitonic emission and decrease in the defect related emission by up to two and one orders of magnitude, respectively, when w is increased in the ∼3-17 nm range. Uniaxial strain build-up within the ZnO NW core with increasing w, as detected by x-ray diffraction measurements, and photocarrier tunneling escape from the ZnO core through the MgO shell enabled by defect-states are proposed as possible mechanisms involved in this effect. These findings are expected to be of key significance for the efficient design and fabrication of ZnO/MgO NW heterostructures and devices.

2.
Nanotechnology ; 27(42): 425501, 2016 Oct 21.
Artículo en Inglés | MEDLINE | ID: mdl-27622391

RESUMEN

We present a glucose biosensor based on ZnO nanowire self-sustained films grown on compacted graphite flakes by the vapor transport method. Nanowire/graphite films were fragmented in water, filtered to form a colloidal suspension, subsequently functionalized with glucose oxidase and finally transferred to a metal electrode (Pt). The obtained devices were evaluated using scanning electron microscopy, energy-dispersive x-ray spectroscopy, cyclic voltammetry and chronoamperometry. The electrochemical responses of the devices were determined in buffer solutions with successive glucose aggregates using a tripolar electrode system. The nanostructured biosensors showed excellent analytical performance, with linear response to glucose concentrations, high sensitivity of up to ≈17 µA cm(-2) mM(-1) in the 0.03-1.52 mM glucose concentration range, relatively low Michaelis-Menten constant, excellent reproducibility and a fast response. The detection limits are more than an order of magnitude lower than those achievable in commercial biosensors for glucose control, which is promising for the development of glucose monitoring methods that do not require blood extraction from potentially diabetic patients. The strong detection enhancements provided by the functionalized nanostructures are much larger than the electrode surface-area increase and are discussed in terms of the physical and chemical mechanisms involved in the detection and transduction processes.

3.
J Appl Microbiol ; 114(3): 762-70, 2013 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-23163324

RESUMEN

AIMS: To investigate the interactions between Botrytis cinerea and other moulds during grape withering and postharvest infection to obtain noble-rotten grapes. METHODS AND RESULTS: Strains of Botrytis cinerea, Penicillium expansum, Penicillium crustosum, Aspergillus niger, Fusarium verticilloides and Alternaria alternata, isolated from naturally withered grapes and identified by molecular tools, were used to infect Garganega and Corvina grapes. Individually sterilized berries were infected by a single inoculation of each strain or a simultaneous inoculation of B. cinerea together with one of each of the other moulds. Withering kinetics, glycerol, gluconic acid, total polyphenols, total anthocyanins and laccase activity greatly varied among each strain and also in respect to untreated berries. Successful noble rot settlement was ascertained by an additional infection assay carried out on nonsterilized berries. CONCLUSIONS: The suitability of inducing noble rot infection during grape withering and the improvement of the health of noble-rotten grapes have been demonstrated. SIGNIFICANCE AND IMPACT OF THE STUDY: This study provides insights on the effects of mould interactions on withered grape quality. Implementing noble rot induction by postharvest infection in winery drying fruit rooms to standardize the level of grape botrytization is encouraged.


Asunto(s)
Botrytis/fisiología , Microbiología de Alimentos , Hongos/fisiología , Vitis/microbiología , Antocianinas/análisis , Botrytis/genética , Hongos/aislamiento & purificación , Genotipo , Gluconatos/análisis , Glicerol/análisis , Lacasa/metabolismo , Vino/microbiología
4.
Radiol Med ; 116(1): 84-91, 2011 Feb.
Artículo en Inglés, Italiano | MEDLINE | ID: mdl-20981500

RESUMEN

PURPOSE: The authors sought to assess the role of arbitration by a third reader of discordant double readings to reduce the rate of recalls to diagnostic assessment. MATERIALS AND METHODS: A consecutive series of 7,660 double readings of screening examinations were considered. Discordant recalls were arbitrated by an expert reader (negative/positive). Diagnostic assessment was performed irrespective of arbitration results, and its outcome was used as reference standard for the study purpose. Assuming that negative arbitration would deny recall, its impact was assessed in terms of reduced recall rate and reduced cancer detection rate. Cost analysis of introducing arbitration was performed according to these results. RESULTS: Recalls at double reading were 528 (6.8%), of which 230 (43.5%) were concordant and 298 (56.5%) were discordant. The latter underwent arbitration, which was negative in 216 (72.4%) and positive in 82 (27.6%) cases, respectively. Overall, 49 cancers were detected (6.39 ‰ screened, 9.2% recalled): 43 cancers were detected among concordant (5.6 ‰ screened, 18.6% concordant) and six among discordant recalls (0.7 ‰ screened, 2.0% discordant). Six cancers were observed among arbitrated cases: five (6%) in positive and one (4.6 ‰) in negative arbitrations. Negative arbitration would have spared 216 assessment procedures (2.8% absolute, 40.9% relative reduction of recall rate) while missing one cancer case (0.13 ‰ absolute, 2.0% relative reduction of cancer detection rate). Arbitration cost was 74 euro, whereas 216 spared assessment procedures would have cost 14,558.4-23,346 euro. CONCLUSIONS: Arbitration is a cost-effective procedure that could be employed as a first measure to counterbalance excess recall rate observed in a double-reading scenario.


Asunto(s)
Neoplasias de la Mama/diagnóstico por imagen , Mamografía , Tamizaje Masivo/normas , Neoplasias de la Mama/patología , Análisis Costo-Beneficio , Errores Diagnósticos/economía , Femenino , Humanos , Italia , Mamografía/economía , Tamizaje Masivo/economía , Negociación , Variaciones Dependientes del Observador , Valor Predictivo de las Pruebas
5.
J Appl Microbiol ; 107(1): 210-8, 2009 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-19245401

RESUMEN

AIMS: To explain the role of Saccharomyces cerevisiae and Saccharomyces uvarum strains (formerly Saccharomyces bayanus var. uvarum) in wine fermentation. METHODS AND RESULTS: Indigenous Saccharomyces spp. yeasts were isolated from Amarone wine (Italy) and analysed. Genotypes were correlated to phenotypes: Melibiose(-) and Melibiose(+) strains displayed a karyotype characterized by three and two bands between 225 and 365 kb, respectively. Two strains were identified by karyotype analysis (one as S. cerevisiae and the other as S. uvarum). The technological characterization of these two strains was conducted by microvinifications of Amarone wine. Wines differed by the contents of ethanol, residual sugars, acetic acid, glycerol, total polysaccharides, ethyl acetate, 2-phenylethanol and anthocyanins. Esterase and beta-glucosidase activities were assayed on whole cells during fermentation at 13 degrees and 20 degrees C. Saccharomyces uvarum displayed higher esterase activity at 13 degrees C, while S. cerevisiae displayed higher beta-glucosidase activity at both temperatures. CONCLUSIONS: The strains differed by important technological and qualitative traits affecting the fermentation kinetics and important aroma components of the wine. SIGNIFICANCE AND IMPACT OF THE STUDY: The contribution of indigenous strains of S. cerevisiae and S. uvarum to wine fermentation was ascertained under specific winemaking conditions. The use of these strains as starters in a winemaking process could differently modulate the wine sensory characteristics.


Asunto(s)
Antocianinas/metabolismo , Esterasas/metabolismo , Etanol/metabolismo , Saccharomyces cerevisiae/metabolismo , Saccharomyces/metabolismo , Vino/microbiología , Ácido Acético/metabolismo , Fermentación , Glicerol/metabolismo , Italia , Cariotipificación , Fenotipo , Compuestos de Amonio Cuaternario/metabolismo , Saccharomyces/genética , Saccharomyces cerevisiae/genética , Saccharomyces cerevisiae/aislamiento & purificación , beta-Glucosidasa/metabolismo
6.
J Clin Endocrinol Metab ; 93(10): 4080-7, 2008 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-18664540

RESUMEN

CONTEXT: Notch genes encode receptors for a signaling pathway that regulates cell growth and differentiation in various contexts, but the role of Notch signaling in thyroid follicular cells has never been fully published. OBJECTIVE: The objective of the study was to characterize the expression of Notch pathway components in thyroid follicular cells and Notch signaling activities in normal and transformed thyrocytes. DESIGN/SETTING AND PATIENTS: Expression of Notch pathway components and key markers of thyrocyte differentiation was analyzed in murine and human thyroid tissues (normal and tumoral) by quantitative RT-PCR and immunohistochemistry. The effects of Notch overexpression in human thyroid cancer cells and FTRL-5 cells were explored with analysis of gene expression, proliferation assays, and experiments involving transfection of a luciferase reporter construct containing human NIS promoter regions. RESULTS: Notch receptors are expressed during the development of murine thyrocytes, and their expression levels parallel those of thyroid differentiation markers. Notch signaling characterized also normal adult thyrocytes and is regulated by TSH. Notch pathway components are variably expressed in human normal thyroid tissue and thyroid tumors, but expression levels are clearly reduced in undifferentiated tumors. Overexpression of Notch-1 in thyroid cancer cells restores differentiation, reduces cell growth rates, and stimulates NIS expression via a direct action on the NIS promoter. CONCLUSION: Notch signaling is involved in the determination of thyroid cell fate and is a direct regulator of thyroid-specific gene expression. Its deregulation may contribute to the loss of differentiation associated with thyroid tumorigenesis.


Asunto(s)
Biomarcadores/metabolismo , Carcinoma Papilar/genética , Diferenciación Celular/genética , Receptores Notch/fisiología , Glándula Tiroides/metabolismo , Neoplasias de la Tiroides/genética , Adenocarcinoma Folicular/genética , Adenocarcinoma Folicular/metabolismo , Animales , Carcinoma Papilar/metabolismo , Desdiferenciación Celular/genética , Células Cultivadas , Regulación hacia Abajo , Regulación Neoplásica de la Expresión Génica , Humanos , Ratones , Especificidad de Órganos/genética , Receptores Notch/genética , Transducción de Señal/genética , Transducción de Señal/fisiología , Simportadores/genética , Simportadores/metabolismo , Glándula Tiroides/citología , Glándula Tiroides/embriología , Neoplasias de la Tiroides/metabolismo
7.
J Clin Endocrinol Metab ; 92(7): 2840-3, 2007 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-17488796

RESUMEN

CONTEXT: BRAF mutations are common in papillary thyroid carcinomas (PTCs). By affecting the expression of genes critically related to the development and differentiation of thyroid cancer, they may influence the prognosis of these tumors. OBJECTIVE: Our objective was to characterize the expression of thyroid-specific genes associated with BRAF mutation in PTCs. DESIGN/SETTING AND PATIENTS: We examined the expression of key markers of thyrocyte differentiation in 56 PTCs with BRAF mutations (BRAF-mut) and 37 with wild-type BRAF (BRAF-wt). Eight samples of normal thyroid tissue were analyzed as controls. Quantitative PCR was used to measure mRNA levels for the sodium/iodide symporter (NIS), apical iodide transporter (AIT-B), thyroglobulin (Tg), thyroperoxidase (TPO), TSH receptor (TSH-R), the transcription factor PAX8, and glucose transporter type 1 (Glut1). NIS protein expression and localization was also analyzed by immunohistochemistry. RESULTS: mRNA levels for all thyroid-specific genes were reduced in all PTCs vs. normal thyroid tissues. NIS, AIT-B, Tg, and TPO expression was significantly lower in BRAF-mut tumors than in the BRAF-wt group. Glut-1 transcript levels were increased in all PTCs, and additional increases were noted in BRAF-mut tumors. In both tumor subsets, the NIS protein that was expressed was abnormally retained in the cytoplasm. CONCLUSION: BRAF V600E mutation in PTCs is associated with reduced expression of key genes involved in iodine metabolism. This effect may alter the effectiveness of diagnostic and/or therapeutic use of radioiodine in BRAF-mut PTCs.


Asunto(s)
Carcinoma Papilar/genética , Regulación Neoplásica de la Expresión Génica , Yodo/metabolismo , Proteínas Proto-Oncogénicas B-raf/genética , Neoplasias de la Tiroides/genética , Adulto , Anciano , Carcinoma Papilar/metabolismo , Diferenciación Celular , Femenino , Marcadores Genéticos , Humanos , Masculino , Persona de Mediana Edad , Mutación Puntual , ARN Mensajero/metabolismo , Neoplasias de la Tiroides/metabolismo
8.
J Clin Invest ; 95(1): 350-9, 1995 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-7814636

RESUMEN

The last exon of the C1-1NH gene was screened for point mutations in 36 unrelated hereditary angioedema patients. Mutations were found in eight patients, predicting changes in the short COOH-terminal region which anchors the reactive site loop on its COOH-terminal side. The effects of each of these mutations were examined in transiently transfected Cos-7 cells. Complete intracellular retention or degradation was observed with substitutions in the COOH-terminal strands 4B or 5B: Leu459-->Pro, Leu459-->Arg, and Pro467-->Arg were all blocked at early stages of intracellular transport, but differences in the immunofluorescence patterns indicated that a significant fraction of the Leu459-->Pro and of the Pro467-->Arg proteins reached a compartment distinct from the endoplasmic reticulum. In line with previous findings with alpha 1-antitrypsin, chain termination within strand 5B resulted in rapid degradation. Mutant Val451-->Met, in strand 1C, and mutant Pro476-->Ser, replacing the invariant proline near the COOH terminus, yielded reduced secretion, but these extracellular proteins were unable to bind the target protease C1s. Presence of low levels of both dysfunctional proteins in patient plasmas defies the conventional classification of C1 inhibitor deficiencies as type I or type II. These data point to a key role of certain residues in the conserved COOH-terminal region of serpins in determining the protein foldings compatible with transport and proper exposure of the reactive site loop.


Asunto(s)
Angioedema/genética , Proteínas Inactivadoras del Complemento 1/genética , Exones/genética , Mutación Puntual , Serpinas/genética , Secuencia de Aminoácidos , Angioedema/clasificación , Secuencia de Bases , Transporte Biológico , Clonación Molecular , Complemento C1/metabolismo , Proteínas Inactivadoras del Complemento 1/metabolismo , Datos de Secuencia Molecular , Reacción en Cadena de la Polimerasa , Unión Proteica , Selección Genética , Análisis de Secuencia de ADN , Homología de Secuencia de Aminoácido , Relación Estructura-Actividad
9.
J Endocrinol Invest ; 30(10): RC31-4, 2007 Nov.
Artículo en Inglés | MEDLINE | ID: mdl-18075281

RESUMEN

There is no effective treatment for recurrent or metastatic medullary thyroid carcinoma (MTC), a tumor arising from thyroid C-cells commonly presenting an inherited or acquired RET mutation. In this study we examined the sensitivity of two human MTC cell lines to novel pyrazolopyrimidine derivates, able to inhibit src-family tyrosine kinase activity. In TT cells [carrying the multiple endocrine neoplasia (MEN)2A Ret mutation Cys 634Trp] and MZ-CRC-1 cells (carrying the MEN2B RET mutation Met891Thr), one of these compounds, namely Si 34, determined a significant growth inhibitory effect (approximately 90% vs control for TT, 80% vs control for MZ-CRC-1) mainly due to enhanced cell mortality after a 6-day incubation. At concentrations that increased cell mortality, neither biochemical or morphological characteristics of apoptosis were detected in TT and MZCRC- 1 cells treated with Si 34. These results, when confirmed in other in vivo preclinical models, suggest that this novel tyrosine kinase inhibitor may be useful for the treatment of MTC.


Asunto(s)
Carcinoma Medular/tratamiento farmacológico , Carcinoma Medular/patología , Pirazoles/farmacología , Pirimidinas/farmacología , Neoplasias de la Tiroides/tratamiento farmacológico , Neoplasias de la Tiroides/patología , Apoptosis/efectos de los fármacos , División Celular/efectos de los fármacos , Línea Celular Tumoral , Humanos , Neoplasia Endocrina Múltiple Tipo 1/tratamiento farmacológico , Neoplasia Endocrina Múltiple Tipo 1/patología , Recurrencia Local de Neoplasia/tratamiento farmacológico , Recurrencia Local de Neoplasia/patología , Poli(ADP-Ribosa) Polimerasas/metabolismo , Inhibidores de Proteínas Quinasas/farmacología , Proteínas Proto-Oncogénicas c-ret/antagonistas & inhibidores
10.
Mol Biol Cell ; 11(1): 355-68, 2000 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-10637313

RESUMEN

Toxoplasma gondii relies on its actin cytoskeleton to glide and enter its host cell. However, T. gondii tachyzoites are known to display a strikingly low amount of actin filaments, which suggests that sequestration of actin monomers could play a key role in parasite actin dynamics. We isolated a 27-kDa tachyzoite protein on the basis of its ability to bind muscle G-actin and demonstrated that it interacts with parasite G-actin. Cloning and sequence analysis of the gene coding for this protein, which we named Toxofilin, showed that it is a novel actin-binding protein. In in vitro assays, Toxofilin not only bound to G-actin and inhibited actin polymerization as an actin-sequestering protein but also slowed down F-actin disassembly through a filament end capping activity. In addition, when green fluorescent protein-tagged Toxofilin was overexpressed in mammalian nonmuscle cells, the dynamics of actin stress fibers was drastically impaired, whereas green fluorescent protein-Toxofilin copurified with G-actin. Finally, in motile parasites, during gliding or host cell entry, Toxofilin was localized in the entire cytoplasm, including the rear end of the parasite, whereas in intracellular tachyzoites, especially before they exit from the parasitophorous vacuole of their host cell, Toxofilin was found to be restricted to the apical end.


Asunto(s)
Actinas/metabolismo , Proteínas de Microfilamentos/metabolismo , Proteínas Protozoarias/metabolismo , Toxoplasma/metabolismo , Proteínas de Capping de la Actina , Animales , Secuencia de Bases , Citosol/metabolismo , ADN Protozoario , Células HeLa , Humanos , Proteínas de Microfilamentos/genética , Datos de Secuencia Molecular , Proteínas Protozoarias/genética , Proteínas Recombinantes de Fusión/genética , Proteínas Recombinantes de Fusión/metabolismo , Toxoplasma/genética
11.
Food Chem ; 213: 378-387, 2016 Dec 15.
Artículo en Inglés | MEDLINE | ID: mdl-27451194

RESUMEN

Amarone wine is different from regular dry wine due to the postharvest withering of Corvina, Corvinone and Rondinella grapes. Grapes were withered in a commercial facility with variability in terms of temperature and relative humidity (R.H.). Sugar content reached 230-240gL(-1) and 280gL(-1) at 20% and 30% mass loss, respectively. Most of VOCs (volatile organic compounds) decreased during withering but few VOCs increased during withering and we considered as markers; in Corvinone they were methylhexanoate, dimethylsuccinate, nerol, nonanoic acid, and benzyl alcohol; in Corvina, benzyl alcohol, isoamyl alcohol, 1-hexanol, p-cymen-8-ol, 2,3 pinanediol, 3-oxo-ionol and 3-methyl-1-pentanol, coumaran and damascenone; in Rondinella, hexanol, nonanoic acid, methyl vanillate, damascenone, 3-oxo-ionol, eugenol, p-cymen-8-ol, 2,3 pinanediol, coumaran and raspberry keton. Olfactive descriptors of the wines and the potential aroma of the combination of Corvina wine with the wines of the other two varieties at different percentages of mass loss are reported.


Asunto(s)
Vitis/química , Compuestos Orgánicos Volátiles/química , Vino/análisis , Olfato , Compuestos Orgánicos Volátiles/análisis
12.
J Mol Biol ; 208(4): 709-14, 1989 Aug 20.
Artículo en Inglés | MEDLINE | ID: mdl-2553984

RESUMEN

The exon-intron structure of the human complement C1s gene displays a striking similarity with that of the gene encoding haptoglobin, a peculiar transport protein distantly related to the serine proteases. While the protease regions of the serine zymogens are typically encoded by multiple exons, the protease domains of C1s and of its genetically linked and functionally interacting homolog C1r are encoded as intronless domains, not unlike a region of haptoglobin, which in fact is devoid of proteolytic activity. The close similarity of the C1s gene with haptoglobin includes the precise conservation of exon-intron junctions and it extends to upstream exons encoding the short repeats typical of several complement components, but found also in other functionally unrelated proteins. Additional evidence of the common ancestry of C1r, C1s and haptoglobin is the presence, within the protease domain, of a set of sequence markers that distinguish these three proteins from all known serine proteases. The finding of vertebrate serine protease genes with an uninterrupted protease-encoding exon supports the definition of a novel evolutionary branch of this gene family and rules out the hypothesis that regards this unusual exon as an irrelevant byproduct of the extravagant functional divergence of haptoglobin.


Asunto(s)
Complemento C1r/genética , Complemento C1s/genética , Haptoglobinas , Serina Endopeptidasas , Secuencia de Aminoácidos , Secuencia de Bases , Exones , Humanos , Intrones , Modelos Genéticos , Datos de Secuencia Molecular
13.
Eur J Cell Biol ; 42(2): 359-64, 1986 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-2434330

RESUMEN

Native tetrameric alpha 2-macroglobulin molecules (alpha 2M) can be converted into a population of dimers by incubation with various divalent cations such as Zn, Cd, Mg, Cu, Ni, Co. This dissociation is completed within 30 min at 37 degrees C. These dimers have a characteristic shape and a size of about 16 X 8 nm, and appear to be the half of the native alpha 2M molecule which has a clear tetrameric structure as seen in the electron microscope. At room temperature or below, dimers obtained with 5 to 100 mM Zn++ can reassociate in long linear polymers which display a regular chain-like arrangement and a helical periodicity. The structural characteristics of this polymer are described. The trypsin inhibitory capacity of Zn++-treated alpha 2M has been studied in an attempt to correlate its Zn++-induced conformational changes with its functional modifications.


Asunto(s)
Inhibidores de Tripsina/antagonistas & inhibidores , Zinc/farmacología , alfa-Macroglobulinas/antagonistas & inhibidores , Cationes Bivalentes/farmacología , Humanos , Sustancias Macromoleculares , Microscopía Electrónica , Conformación Proteica/efectos de los fármacos
14.
Hypertension ; 7(6 Pt 1): 995-1002, 1985.
Artículo en Inglés | MEDLINE | ID: mdl-4077227

RESUMEN

This study was undertaken to test whether the emphasized systemic vasomotion during sympathetic activation in hypertension is shared by the pulmonary circulation. To this end, 10 normotensive and 29 primary hypertensive subjects were investigated during adrenergic stimulation by mental arithmetic and cold pressor test. Both stimuli induced a systemic pressor reaction in both groups, which was mediated through an increase in cardiac output and a mild reduction in vascular resistance during arithmetic and through a predominant rise in systemic vascular resistance during cold. Each of these changes was emphasized in the hypertensive population as compared with the normotensive one. Pressure in the pulmonary artery remained unchanged during cold and was slightly raised (systolic) during arithmetic in normotensive subjects. On the contrary, in hypertensive subjects systolic and diastolic pulmonary pressures were consistently augmented by both stimuli, and pulmonary arteriolar resistance (dyn sec cm-5) rose from 92 in the baseline to 125 (p less than 0.01) during arithmetic and to 124 (p less than 0.01) during the cold test. This reaction is interpreted as reflecting a neurally mediated vasoconstriction and not as the consequence of mechanical or chemical changes, since no difference was observed in pulmonary wedge pressure, pleural pressure, arterial blood gas levels, and pH between controls and hypertensive subjects in the steady state and during either stressful stimulation. Baseline pulmonary arteriolar resistance was also found to correlate positively with systemic vascular resistance in the hypertensive group. When pressure changes occurred, the time course was similar in the two circuits; resistance increased to a proportionally similar degree in the two districts during the cold stimulus.(ABSTRACT TRUNCATED AT 250 WORDS)


Asunto(s)
Hipertensión/fisiopatología , Circulación Pulmonar , Adulto , Presión Sanguínea , Gasto Cardíaco , Frío , Femenino , Humanos , Masculino , Procesos Mentales/fisiología , Persona de Mediana Edad , Arteria Pulmonar/fisiopatología , Resistencia Vascular
15.
Eur J Cancer ; 32A(3): 498-505, 1996 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-8814698

RESUMEN

The immunogenicity of the idiotypic portions of two antigrowth factor receptor monoclonal antibodies (MAbs) was studied. Immunisation of allogeneic but not syngeneic mice with antihuman epidermal growth factor receptor (EGF-R) MAb MINT5 or anti-HER-2/neu MGR6 MAb elicited a detectable titre of circulating antibodies, particularly when the MAb was coupled with the keyhole limpet haemocyanin and administered together with Freund's adjuvant. The anti-Ab1 response to MAb MINT5 was slightly delayed as compared with the response obtained with MAb MGR6 and was mainly directed to the variable regions. In both cases, all anti-Ab1-positive sera specifically competed with the binding of homologous radiolabelled Ab1 to the relevant EGF-R+ or HER-2/neu+ target cells. Fusion of splenocytes from MINT5-immunised animals failed to produce MAb, whereas cell fusion was successful in generating a paratope-related MAb in the case of MGR6. The anti-MGR6 MAb-produced IdM6.4 inhibited the binding of MAb MGR6 on breast carcinoma cells, suggesting that it recognises an idiotope in or near the antigen combining site, and can be considered useful in the identification and purification of the Ab1 or its derivatives. We analysed whether a possible recognition of murine EGF-R by MAb MINT5 or a mimicry of EGF by the MAb idiotype prevented or delayed the development of an idiotypic cascade in mice. MINT5 inhibited human and murine EGF binding to the human EGF-R, whereas the anti-Ab1 response competed with MINT5 but not with murine EGF binding to A431 human epidermoid carcinoma cells. Moreover, MINT5 did not recognise the murine EGF-R. In a phase I clinical study, no detectable levels of human antimouse antibody response were observed in 5 of the 6 treated cancer patients. The ability of MAb MINT5 to block human EGF-R function, together with its low immunogenicity in patients, raise the possibility of its application in carcinoma immunotherapy.


Asunto(s)
Anticuerpos Antiidiotipos/sangre , Anticuerpos Monoclonales/inmunología , Receptores de Factores de Crecimiento/inmunología , Animales , Anticuerpos Monoclonales/biosíntesis , Anticuerpos Monoclonales/farmacología , Especificidad de Anticuerpos , Neoplasias de la Mama/inmunología , Femenino , Humanos , Neoplasias Pulmonares/inmunología , Ratones , Ratones Endogámicos BALB C , Ratones Endogámicos C57BL , Neoplasias Ováricas/inmunología , Células Tumorales Cultivadas
16.
J Hypertens ; 3(1): 39-45, 1985 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-2987340

RESUMEN

We have examined the mechanisms mediating the release of renin elicited in man by reduction of renal perfusion pressure. Fifteen patients with essential hypertension and six normotensive subjects were investigated during diagnostic renal arteriography. Renal neural receptors were inhibited by propranolol (10 mg i.v.) and activated by a standard cold pressor test. Vascular receptors were stimulated by unilateral reduction of renal perfusion pressure by 50%, using a balloon-tipped catheter. The stimulus caused release of renin. In hypertensives, arterial plasma renin increased by 44, 69 and 73% of control at 5, 15 and 30 min, respectively. Adrenergic activation by cold raised the arterial and the renal venous renin by approximately 50% of control and caused a fourfold rise when it was combined with the arterial obstruction. Following propranolol the renin response to reduction of the renal perfusion pressure was delayed and reduced, and cold stimulation, both alone and in combination with arterial obstruction, failed to stimulate renin release. Findings were qualitatively and quantitatively similar in the normotensive group. This study supports the hypothesis that the renin response to reduction of renal perfusion pressure in man results from an interaction of adrenergic and vascular receptors. It cannot be stated whether the former are synergistic or supplementary to the latter, even though adrenergic activation by cold stimulation provides evidence that a synergism between the two may exist.


Asunto(s)
Hipertensión/fisiopatología , Aparato Yuxtaglomerular/metabolismo , Presorreceptores/fisiología , Receptores Adrenérgicos beta/fisiología , Circulación Renal , Renina/metabolismo , Adulto , Frío , Femenino , Humanos , Hipertensión Renovascular/fisiopatología , Masculino , Persona de Mediana Edad , Perfusión , Presorreceptores/efectos de los fármacos , Propranolol/farmacología , Receptores Adrenérgicos beta/efectos de los fármacos
17.
Mol Cell Endocrinol ; 49(2-3): 173-9, 1987 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-3556752

RESUMEN

The major protein synthesized in vitro by the ovine thyroid cell line OVNIS 6H is the prothyroid hormone thyroglobulin. Purified from serum-free cell culture media using sucrose gradient centrifugation, the thyroglobulin dimer was analysed for iodine content and observed by electron microscopy. In their usual medium, the OVNIS 6H cells produce a very poorly iodinated thyroglobulin containing 0.05 I atom per molecule. When cultured with methimazole or propylthiouracil, two inhibitors of iodide organification, less than 0.007 I atom/molecules was found. These molecules purified from cell cultures were compared to those purified from ovine thyroid tissue containing 26 I atoms/mol. Despite large differences in iodine content, the three preparations all consist of 19 S thyroglobulin dimers with the classical ovoidal shape. The variability in size measurements remains in a 2% range for all thyroglobulin types. Consequently, no real significant variation can be found between the highly iodinated thyroglobulin isolated from tissue, and the poorly or non-iodinated thyroglobulins isolated from cells cultured with or without methimazole or propylthiouracil.


Asunto(s)
Tiroglobulina , Animales , Células Cultivadas , Yodo , Sustancias Macromoleculares , Microscopía Electrónica , Conformación Proteica , Ovinos , Relación Estructura-Actividad
18.
Tumori ; 78(1): 1-4, 1992 Feb 29.
Artículo en Inglés | MEDLINE | ID: mdl-1609451

RESUMEN

The production and characterization of two new monoclonal antibodies (MAbs), designated MAR4 and MAR5, raised against the partially purified alpha 5 beta 1 integrin, are described. The reactivity of these 2 MAbs on tumor cell lines indicated that they reacted on all the cells expressing the beta 1 subunit independently of the alpha 5 expression. Both MAbs were found to immunoprecipitate on 3 cell lines, a protein of 120 KD corresponding to the molecular weight be the beta 1 chain, in addition to proteins of other MW corresponding to the alpha subunits differentially expressed by these cells. The cross-competition experiments showed that MAR4 and MAR5 recognize the same epitope. These 2 MAbs seem to be useful reagents for the characterization of the VLA expression in tumor cells.


Asunto(s)
Anticuerpos Monoclonales/biosíntesis , Integrinas/inmunología , Animales , Anticuerpos Monoclonales/análisis , Anticuerpos Monoclonales/aislamiento & purificación , Unión Competitiva , Línea Celular , Técnica del Anticuerpo Fluorescente , Humanos , Hibridomas/inmunología , Radioisótopos de Yodo , Ratones , Ratones Endogámicos BALB C , Pruebas de Precipitina , Células Tumorales Cultivadas/inmunología
19.
Angiology ; 38(9): 672-9, 1987 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-3310745

RESUMEN

Excessive vascular tone and overresponsiveness to adrenergic stimuli characterize the hemodynamics of the greater and the lesser circulation in hypertension. We tested whether calcium entry blockade with verapamil (11 cases) or nifedipine (11 cases) may improve the vascular regulation in high blood pressure. Mental arithmetic and cold were used as adrenergic activators. The former stimulus produced obvious elevation of epinephrine plasma concentration, increase of cardiac output (CO), slight systemic vasodilatation, pulmonary vasoconstriction, and rise of blood pressure in both circuits. After calcium antagonists, the epinephrine reaction to the arithmetic test was significantly attenuated, variations in CO and systemic blood pressure were unchanged, pulmonary vasoconstriction was abolished, and the pressure rise in the lesser circuit was halved. The cold pressor test increased norepinephrine plasma concentration (NE pc), systemic and pulmonary blood pressure, and vascular resistance and did not alter CO. The attained NE pc during cold was unvaried after verapamil and significantly enhanced after nifedipine; pressure and resistance responses of the two circuits were almost unchanged after the former, whereas systemic and pulmonary vascular resistance rises were importantly attenuated after the latter compound, resulting in much lower pressure reactivity. A modulation of the sympathoadrenal reaction, per se, can explain changes in the systemic and in the pulmonary vasomotion with calcium blockade during arithmetic. It would seem that after verapamil the sympathetic system was still activated during cold to such an extent as to maintain the same vasoconstrictor potency. NE pc suggests that the sympathetic discharge was not reduced by nifedipine.(ABSTRACT TRUNCATED AT 250 WORDS)


Asunto(s)
Circulación Sanguínea/efectos de los fármacos , Hipertensión/tratamiento farmacológico , Nifedipino/farmacología , Circulación Pulmonar/efectos de los fármacos , Resistencia Vascular/efectos de los fármacos , Verapamilo/farmacología , Adulto , Presión Sanguínea/efectos de los fármacos , Ensayos Clínicos como Asunto , Cognición/efectos de los fármacos , Frío , Hemodinámica/efectos de los fármacos , Humanos , Masculino , Persona de Mediana Edad , Nifedipino/uso terapéutico , Norepinefrina/sangre , Estudios Prospectivos , Distribución Aleatoria , Verapamilo/uso terapéutico
20.
Hybridoma ; 11(4): 437-46, 1992 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-1383124

RESUMEN

In order to improve the purification of immunoconjugates containing alpha sarcin, a ribosome-inactivating protein, and in the attempt to define the enzymic region of the toxin, MAbs against alpha sarcin were produced. From 5 fusions, by adopting a short period of immunization and very low doses of the immunogen, 10 anti-toxin-producing clones were obtained. One of them, named MAsg2 (IgG2b), due to its specific reactivity and secreting properties, was selected for further characterization. MAsg2 was found to recognize an epitope which is common to two, i.e. alpha sarcin and clavatin, of the three different aspergillins tested, but is not involved in the active site of the toxins.


Asunto(s)
Anticuerpos Monoclonales/biosíntesis , Endorribonucleasas , Proteínas Fúngicas/inmunología , Animales , Anticuerpos Monoclonales/aislamiento & purificación , Antígenos Fúngicos , Aspergillus , Epítopos , Proteínas Fúngicas/toxicidad , Hibridomas/inmunología , Inmunización , Inmunotoxinas/aislamiento & purificación , Ratones , Inhibidores de la Síntesis de la Proteína/inmunología , Inhibidores de la Síntesis de la Proteína/toxicidad , Ribosomas/efectos de los fármacos
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