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1.
Mass Spectrom Rev ; 42(4): 1082-1083, 2023.
Artículo en Inglés | MEDLINE | ID: mdl-34854482
2.
Chem Senses ; 41(2): 177-86, 2016 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-26708734

RESUMEN

Mandrills are one of the few Old World primates to show scent-marking. We combined ethological and chemical approaches to improve our understanding of this behavior in 3 zoo-managed groups. We observed the olfactory behavior performed by adults and adolescents (N = 39) for 775h. We investigated the volatile components of sternal scent-marks using gas chromatography-mass spectrometry and compared volatile profiles with traits of the signaler. Males marked more than females and within each sex the frequency of scent-marking was related to age and dominance status, but alpha males scent-marked most frequently and particularly in specific areas at the enclosure boundaries. We identified a total of 77 volatile components of sternal gland secretion, including compounds functioning as male sex pheromones in other mammals, in scent-marks spontaneously released on filter paper by 27 male and 18 female mandrills. We confirmed our previous findings that chemical profiles contain information including sex, male age and rank, and we also found that odor may encode information about group membership in mandrills. Our results support the hypotheses that scent-marking signals the status of the dominant male as well as playing territorial functions but also suggest that it is part of sociosexual communication.


Asunto(s)
Envejecimiento , Comunicación Animal , Animales de Zoológico/fisiología , Jerarquia Social , Mandrillus/fisiología , Odorantes , Glándulas Odoríferas/metabolismo , Factores de Edad , Animales , Femenino , Cromatografía de Gases y Espectrometría de Masas , Procesos de Grupo , Masculino , Factores Sexuales , Territorialidad
3.
Clin Sci (Lond) ; 130(20): 1793-806, 2016 10 01.
Artículo en Inglés | MEDLINE | ID: mdl-27439970

RESUMEN

Berberine (BRB) is commonly used in herbal medicine, but its mechanisms of action are poorly understood. In the present study, we tested BRB in steatohepatitis induced by a methionine- and choline-deficient (MCD) diet, in acute acetaminophen intoxication and in cultured murine macrophages. BRB markedly improved parameters of liver injury and necroinflammation induced by the MCD diet, although increased mortality was observed by mechanisms independent of bacterial infections or plasma levels of BRB. The MCD diet induced up-regulation of all components of the NLRP3 (NACHT, LRR and PYD domain-containing protein 3) inflammasome, and increased hepatic levels of mature IL-1ß (interleukin 1ß). All of these parameters were significantly reduced in mice treated with BRB. In mice administered an acetaminophen overdose, a model dependent on inflammasome activation, BRB reduced mortality and ALT (alanine aminotransferase) elevation, and limited the expression of inflammasome components. In vitro, LPS (lipopolysaccharide)-induced activation of NLRP3 inflammasome in RAW264.7 murine macrophages was markedly decreased by pre-incubation with BRB. BRB significantly limited the activation of the purinergic receptor P2X7, involved in the late phases of inflammasome activation. Upon P2X7 knockdown, the ability of BRB to block LPS-induced secretion of IL-1ß was lost. These data indicate that administration of BRB ameliorates inflammation and injury in two unrelated murine models of liver damage. We demonstrate for the first time that BRB interferes with activation of the NLRP3 inflammasome pathway in vivo and in vitro, through a mechanism based on interference with activation of P2X7, a purinergic receptor involved in inflammasome activation.


Asunto(s)
Acetaminofén/efectos adversos , Berberina/administración & dosificación , Enfermedad Hepática Inducida por Sustancias y Drogas/tratamiento farmacológico , Inflamasomas/metabolismo , Animales , Enfermedad Hepática Inducida por Sustancias y Drogas/genética , Enfermedad Hepática Inducida por Sustancias y Drogas/metabolismo , Humanos , Inflamasomas/efectos de los fármacos , Inflamasomas/genética , Interleucina-1beta/genética , Interleucina-1beta/metabolismo , Macrófagos/efectos de los fármacos , Macrófagos/metabolismo , Masculino , Ratones , Ratones Endogámicos C57BL , Proteína con Dominio Pirina 3 de la Familia NLR/antagonistas & inhibidores , Proteína con Dominio Pirina 3 de la Familia NLR/genética , Proteína con Dominio Pirina 3 de la Familia NLR/metabolismo , Transducción de Señal
4.
Eur J Mass Spectrom (Chichester) ; 19(6): 475-82, 2013.
Artículo en Inglés | MEDLINE | ID: mdl-24378465

RESUMEN

During the last decade, significant technological improvements in mass spectrometry have had a great impact on drug discovery. The development of matrix-assisted laser desorption/ionization imaging mass spectrometry (MALDI-IMS) has set a new frontier for the study of the distribution of endogenous and exogenous molecules present within a tissue. MALDI-IMS is a surface sampling technique that allows not only the detection of multiple analytes but also gives the spatial distribution of those analytes. Active compounds for pulmonary disease need an optimal and well-studied delivery into the lungs, in order to assure distribution with greater penetration into the peripheral or the alveolar region of the lung to maximize the therapeutic effects. IMS is very useful in the field of drug discovery, showing drug delivery and distribution in the body and organs. In this study, we present a comparison between two different ways of carrying out pulmonary drug administration: inhalation of a nebulized aerosol of aqueous drug solutions and intratracheal administration, which is much simpler, not expensive and commonly used during in vivo screening. Tiotropium bromide is a long-acting anticholinergic medicine used for maintenance treatment of chronic obstructive pulmonary disease. In the present work, tiotropium was administered by nebulization and by intratracheal instillation to guinea pigs at doses able to induce significant anti-bronchoconstrictive activity. Lung samples were dissected, frozen, cryosectioned and coated with matrix (α-hydroxy-cinnamic acid). IMS analyses were performed using a MALDI-LTQ-Orbitrap XL. Using this technique we were able to compare different distributions of the drug depending on the method of administration.


Asunto(s)
Sistemas de Liberación de Medicamentos/métodos , Pulmón/metabolismo , Enfermedad Pulmonar Obstructiva Crónica/tratamiento farmacológico , Derivados de Escopolamina/farmacocinética , Espectrometría de Masa por Láser de Matriz Asistida de Ionización Desorción/métodos , Administración por Inhalación , Aerosoles , Animales , Antagonistas Colinérgicos/farmacocinética , Vías de Administración de Medicamentos , Descubrimiento de Drogas , Cobayas , Masculino , Nebulizadores y Vaporizadores , Enfermedad Pulmonar Obstructiva Crónica/metabolismo , Bromuro de Tiotropio , Distribución Tisular
5.
J Allergy Clin Immunol ; 129(5): 1314-1320.e3, 2012 May.
Artículo en Inglés | MEDLINE | ID: mdl-22104605

RESUMEN

BACKGROUND: Treatment with aqueous and aluminum hydroxide (Al[OH](3))-adsorbed purified honeybee (Apis mellifera) venom (HBV) preparations can reduce the incidence of side effects associated with venom immunotherapy. OBJECTIVE: The aim of the present study was to assess these purified HBV immunotherapy preparations in situ. METHODS: Matrix-assisted laser desorption/ionization mass spectrometry imaging (MALDI-MSI) was used to visualize the distribution of HBV components. The preparations were administered on the back legs of naive Wistar rats. The rats were killed, and cryosectioned tissue sections were subjected to hematoxylin and eosin staining and MALDI-MSI analyses. RESULTS: Low-density maps of tissue distribution of HBV peptides, such as secapin, mast cell degranulating peptide, and melittin (Api m 4) were detected in the tissue after administration of HBV immunotherapy preparations. In addition, release of biogenic amines, cytokines, and leukotrienes was observed, and the distribution of HBV allergens, such as Api m 1 and Api m 2, was shown. At the 24-hour time point, the major HBV allergen Api m 1 was still detected at the site of Al(OH)(3)-adsorbed HVB injection, whereas in the case of aqueous HBV preparation, all the allergens, as well as most of the biogenic amines, were cleared at the 24-hour time point. CONCLUSION: The present study shows that the majority of low-molecular-weight HBV components are rapidly removed from the site of venom immunotherapy administration. Furthermore, Al(OH)(3)-adsorbed HBV preparation demonstrated a depot effect, prolonging the availability of bee venom allergens at the site of administration.


Asunto(s)
Venenos de Abeja/inmunología , Desensibilización Inmunológica , Hipersensibilidad/tratamiento farmacológico , Hipersensibilidad/inmunología , Alérgenos/administración & dosificación , Alérgenos/efectos adversos , Alérgenos/farmacocinética , Hidróxido de Aluminio/administración & dosificación , Hidróxido de Aluminio/química , Animales , Antígenos de Plantas/administración & dosificación , Antígenos de Plantas/efectos adversos , Venenos de Abeja/efectos adversos , Venenos de Abeja/metabolismo , Abejas , Aminas Biogénicas/metabolismo , Crioultramicrotomía , Humanos , Hialuronoglucosaminidasa/administración & dosificación , Hialuronoglucosaminidasa/efectos adversos , Hialuronoglucosaminidasa/farmacocinética , Hipersensibilidad/diagnóstico , Proteínas de Insectos/administración & dosificación , Proteínas de Insectos/efectos adversos , Proteínas de Insectos/farmacocinética , Rayos Láser/estadística & datos numéricos , Meliteno/efectos adversos , Meliteno/inmunología , Péptidos/metabolismo , Fosfolipasas A/administración & dosificación , Fosfolipasas A/efectos adversos , Fosfolipasas A/farmacocinética , Ratas , Ratas Wistar , Espectrometría de Masa por Láser de Matriz Asistida de Ionización Desorción/instrumentación , Espectrometría de Masa por Láser de Matriz Asistida de Ionización Desorción/métodos , Agua/administración & dosificación , Agua/química
6.
Anal Bioanal Chem ; 404(10): 2927-38, 2012 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-23064671

RESUMEN

Cardiovascular diseases are the world's number one cause of death, accounting for 17.1 million deaths a year. New high-resolution molecular and structural imaging strategies are needed to understand underlying pathophysiological mechanism. The aim of our study is (1) to provide a molecular basis of the heart animal model through the local identification of biomolecules by mass spectrometry imaging (MSI) (three-dimensional (3D) molecular reconstruction), (2) to perform a cross-species validation of secondary ion mass spectrometry (SIMS)-based cardiovascular molecular imaging, and (3) to demonstrate potential clinical relevance by the application of this innovative methodology to human heart specimens. We investigated a MSI approach using SIMS on the major areas of a rat and mouse heart: the pericardium, the myocardium, the endocardium, valves, and the great vessels. While several structures of the heart can be observed in individual two-dimensional sections analyzed by metal-assisted SIMS imaging, a full view of these structures in the total heart volume can be achieved only through the construction of the 3D heart model. The images of 3D reconstruction of the rat heart show a highly complementary localization between Na(+), K(+), and two ions at m/z 145 and 667. Principal component analysis of the MSI data clearly identified different morphology of the heart by their distinct correlated molecular signatures. The results reported here represent the first 3D molecular reconstruction of rat heart by SIMS imaging.


Asunto(s)
Corazón/anatomía & histología , Imagenología Tridimensional/métodos , Imagen Molecular/métodos , Miocardio/ultraestructura , Espectrometría de Masa de Ion Secundario/métodos , Animales , Humanos , Ratones , Análisis de Componente Principal , Ratas , Programas Informáticos
7.
J Biol Chem ; 285(44): 34106-14, 2010 Oct 29.
Artículo en Inglés | MEDLINE | ID: mdl-20724478

RESUMEN

The NAD rescue pathway consists of two enzymatic steps operated by nicotinamide phosphoribosyltransferase (Nampt) and nicotinamide mononucleotide adenylyltransferases. Recently, the potent Nampt inhibitor FK866 has been identified and evaluated in clinical trials against cancer. Yet, how Nampt inhibition affects NAD contents and bioenergetics is in part obscure. It is also unknown whether NAD rescue takes place in mitochondria, and FK866 alters NAD homeostasis within the organelle. Here, we show that FK866-dependent reduction of the NAD contents is paralleled by a concomitant increase of ATP in various cell types, in keeping with ATP utilization for NAD resynthesis. We also show that poly- and mono(ADP-ribose) transferases rather than Sirt-1 are responsible for NAD depletion in HeLa cells exposed to FK866. Mass spectrometry reveals that the drug distributes in the cytosolic and mitochondrial compartment. However, the cytoplasmic but not the mitochondrial NAD pool is reduced upon acute or chronic exposure to the drug. Accordingly, Nampt does not localize within the organelles and their bioenergetics is not affected by the drug. In the mouse, FK866-dependent reduction of NAD contents in various organs is prevented by inhibitors of poly(ADP-ribose) polymerases or the NAD precursor kynurenine. For the first time, our data indicate that mitochondria lack the canonical NAD rescue pathway, broadening current understanding of cellular bioenergetics.


Asunto(s)
Inhibidores Enzimáticos/farmacología , Mitocondrias/metabolismo , Nicotinamida Fosforribosiltransferasa/antagonistas & inhibidores , Acrilamidas/farmacología , Adenosina Trifosfato/química , Animales , Fibroblastos/metabolismo , Células HeLa , Humanos , Quinurenina/química , Masculino , Ratones , NAD/química , Piperidinas/farmacología , Poli(ADP-Ribosa) Polimerasas/metabolismo
8.
Hepatology ; 52(2): 493-505, 2010 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-20683949

RESUMEN

UNLABELLED: Antidiabetic thiazolidinediones (TZD) have in vitro antiproliferative effect in epithelial cancers, including hepatocellular carcinoma (HCC). The effective anticancer properties and the underlying molecular mechanisms of these drugs in vivo remain unclear. In addition, the primary biological target of TZD, the ligand-dependent transcription factor peroxisome proliferator-activated receptor gamma (PPARgamma), is up-regulated in HCC and seems to provide tumor-promoting responses. The aim of our study was to evaluate whether chronic administration of TZD may affect hepatic carcinogenesis in vivo in relation to PPARgamma expression and activity. The effect of TZD oral administration for 26 weeks was tested on tumor formation in PPARgamma-expressing and PPARgamma-deficient mouse models of hepatic carcinogenesis. Proteomic analysis was performed in freshly isolated hepatocytes by differential in gel electrophoresis and mass spectrometry analysis. Identified TZD targets were confirmed in cultured PPARgamma-deficient hepatocytes. TZD administration in hepatitis B virus (HBV)-transgenic mice (TgN[Alb1HBV]44Bri) reduced tumor incidence in the liver, inhibiting hepatocyte proliferation and increasing apoptosis. PPARgamma deletion in hepatocytes of HBV-transgenic mice (Tg[HBV]CreKOgamma) did not modify hepatic carcinogenesis but increased the TZD antitumorigenic effect. Proteomic analysis identified nucleophosmin (NPM) as a TZD target in PPARgamma-deficient hepatocytes. TZD inhibited NPM expression at protein and messenger RNA levels and decreased NPM promoter activity. TZD inhibition of NPM was associated with the induction of p53 phosphorylation and p21 expression. CONCLUSION: These findings suggest that chronic administration of TZD has anticancer activity in the liver via inhibition of NPM expression and indicate that these drugs might be useful for HCC chemoprevention and treatment.


Asunto(s)
Carcinoma Hepatocelular/prevención & control , Carcinoma Hepatocelular/virología , Virus de la Hepatitis B , Neoplasias Hepáticas/prevención & control , Neoplasias Hepáticas/virología , Proteínas Nucleares/fisiología , PPAR gamma/fisiología , Tiazolidinedionas/uso terapéutico , Animales , Hepatocitos , Ratones , Ratones Transgénicos , Neoplasias Experimentales/prevención & control , Neoplasias Experimentales/virología , Nucleofosmina , Tiazolidinedionas/farmacología , Células Tumorales Cultivadas
9.
Chem Senses ; 36(4): 335-44, 2011 May.
Artículo en Inglés | MEDLINE | ID: mdl-21220518

RESUMEN

The genome of the silkmoth Bombyx mori contains 44 genes encoding odorant-binding proteins (OBPs) and 20 encoding chemosensory proteins (CSPs). In this work, we used a proteomic approach to investigate the expression of proteins of both classes in the antennae of adults and in the female pheromone glands. The most abundant proteins found in the antennae were the 4 OBPs (PBP, GOBP1, GOBP2, and ABP) and the 2 CSPs (CSP1 and CSP2) previously identified and characterized. In addition, we could detect only 3 additional OBPs and 2 CSPs, with clearly different patterns of expression between the sexes. Particularly interesting, on the other hand, is the relatively large number of binding proteins (1 OBP and 7 CSPs) expressed in the female pheromone glands, some of them not present in the antennae. In the glands, these proteins could be likely involved in the solubilization of pheromonal components and their delivery in the environment.


Asunto(s)
Bombyx/metabolismo , Proteínas de Insectos/metabolismo , Feromonas/metabolismo , Receptores Odorantes/metabolismo , Animales , Antenas de Artrópodos/metabolismo , Bombyx/genética , Femenino , Expresión Génica , Proteínas de Insectos/genética , Masculino , Proteómica , Receptores Odorantes/genética , Espectrometría de Masa por Láser de Matriz Asistida de Ionización Desorción
10.
Rapid Commun Mass Spectrom ; 25(20): 3091-6, 2011 Oct 30.
Artículo en Inglés | MEDLINE | ID: mdl-21953964

RESUMEN

1,5-Diaminonaphthalene (DAN) has been described as an interesting and effective matrix for matrix-assisted laser desorption/ionization (MALDI) experiments in positive ion mode, being able to activate in-source decomposition phenomena and, when employed for the analysis of proteins containing disulphide bridge(s), being able to activate reduction processes, resulting in disulphide bridge cleavage. The mechanisms of the DAN reactivity have been studied in detail, and the results indicate that the reduction properties of the matrix are of a radical nature. In the present study the structure of the reactive species produced by DAN, responsible for its reductive properties, has been investigated by accurate mass measurements and tandem mass spectrometry (MS/MS) experiments. Contrary to what is usually observed by laser irradiation of other MALDI matrices (with the sole formation of the MH(+) ion of the matrix), DAN leads to the formation of odd-electron molecular ions M(+•) . This can be rationalized by the occurrence of two photon pooling processes, due to the low ionization energy of DAN. Thus the M(+•) ion of DAN can be considered responsible for both analyte protonation and disulphide bond reduction and some mechanisms are proposed for this behaviour.


Asunto(s)
2-Naftilamina/análogos & derivados , Espectrometría de Masa por Láser de Matriz Asistida de Ionización Desorción/métodos , 2-Naftilamina/química , Disulfuros/química , Depuradores de Radicales Libres , Insulina/química , Iones/química , Oxidación-Reducción , Proteínas/química , Protones , Espectrometría de Masa por Láser de Matriz Asistida de Ionización Desorción/instrumentación
11.
J Proteome Res ; 9(4): 1822-33, 2010 Apr 05.
Artículo en Inglés | MEDLINE | ID: mdl-20155982

RESUMEN

Chemical communication in insects is mediated by soluble binding proteins, belonging to two large families, Odorant-binding Proteins (OBPs) and Chemosensory Proteins (CSPs). Recently, evidence has been provided that OBPs are involved in recognition of chemical stimuli. We therefore decided to investigate the expression of OBPs and CSPs in the honeybee at the protein level, using a proteomic approach. Our results are in agreement with previous reports of expression at the RNA level and show that 12 of the 21 OBPs predicted in the genome of the honeybee Apis mellifera and 2 of the 6 CSPs are present in the foragers' antennae, while the larvae express only three OBPs and a single CSP. MALDI mass spectrometry on crude antennal extracts and MALDI profiling on sections of antennae demonstrated that these techniques can be applied to investigate individual differences in the expression of abundant proteins, such as OBPs and CSPs, as well as to detect the presence of proteins in different regions of the antenna. Finally, as part of a project aimed at the characterization of all OBPs and CSPs of the honeybee, we expressed 5 OBPs and 4 CSPs in a bacterial system and measured their affinity to a number of ligands. Clear differences in their binding spectra have been observed between OBPs, as well as CSPs.


Asunto(s)
Abejas/metabolismo , Proteínas de Insectos/biosíntesis , Receptores Odorantes/química , Animales , Abejas/genética , Electroforesis en Gel Bidimensional , Perfilación de la Expresión Génica/métodos , Proteínas de Insectos/clasificación , Proteínas de Insectos/genética , Proteínas de Insectos/metabolismo , Proteómica/métodos , Espectrometría de Masa por Láser de Matriz Asistida de Ionización Desorción
12.
Bioconjug Chem ; 21(8): 1432-8, 2010 Aug 18.
Artículo en Inglés | MEDLINE | ID: mdl-20669938

RESUMEN

Immunotherapy of tumors and of melanoma in particular has a long history, and recently this therapeutic approach found a reliable scientific rationale. This biological therapy aims to teach the patient's immune system to recognize the antigens expressed on tumor cells and destroy them, leaving normal cells intact. The success of this therapy highly depends on the selection of target antigens that are essential for tumors growth and progression. The overexpression of GM(3) ganglioside 1 and especially the expression of its metabolite GM(3) lactone 2 characterize murine and human melanomas, playing an important role in tumor progression and making such self-antigens potential targets for the immunotherapy of these neoplasms. Although more immunogenic than its precursor, GM(3) lactone 2 is unsuitable to be used in immunotherapy as a melanoma-associated antigen (MAA) because it is unstable under physiological conditions. We designed and synthesized the hydrolytically stable mimetic 3, which is remarkably simpler than the native lactone 2; after conjugation of 3 to the protein carrier keyhole-limpet hemocyanin (KLH), the obtained glycoprotein 5 was used as the immunogen in vivo to successfully elicit specific antimelanoma antibodies. In fact, no appreciable binding to GM(1) was observed. Capitalizing on the stability and on the reduced structural complexity of mimetic 3, the immunostimulant 5 we report represents a new promising synthetic glycoconjugate for the immunotherapy of melanoma.


Asunto(s)
Anticuerpos/inmunología , Gangliósido G(M3)/análogos & derivados , Melanoma/inmunología , Imitación Molecular/inmunología , Animales , Anticuerpos/química , Especificidad de Anticuerpos , Reacciones Antígeno-Anticuerpo , Conformación de Carbohidratos , Simulación por Computador , Gangliósido G(M3)/química , Gangliósido G(M3)/genética , Gangliósido G(M3)/inmunología , Hemocianinas/química , Hemocianinas/inmunología , Humanos , Inmunoterapia , Melanoma/genética , Melanoma/terapia , Ratones , Imitación Molecular/genética
13.
Neurol Sci ; 31(3): 283-91, 2010 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-19936883

RESUMEN

The aim of our pilot study was to investigate, by a proteomic approach, the expressed differences in cerebrospinal fluid (CSF) protein patterns in order to aid in the diagnosis and treatment of normal pressure hydrocephalus (NPH). Seventeen patients with NPH, selected by Intracranial-Pressure monitoring (ICPmo), underwent implantation of a shunt and after 6 months were clinically re-evaluated. Thirteen patients improved, whereas four did not. During ICPmo CSF was collected and its proteoma was analyzed by 2D gel electrophoresis and mass spectrometry. The over-expression of alpha2HS glycoprotein, alpha1 antichimotrypsin and alpha1beta glycoprotein and the under-expression of glial fibrillary acidic protein, apolipoproteins (AIV, J and E), complement C3c, anti-thrombin, alpha2 antiplasmin and albumin seem to be associated with a positive response to surgery. Most of these proteins have been reported to be altered in Alzheimer disease, supporting the hypothesis of a possible link between these two nosological entities.


Asunto(s)
Derivaciones del Líquido Cefalorraquídeo , Hidrocéfalo Normotenso/líquido cefalorraquídeo , Hidrocéfalo Normotenso/cirugía , Anciano , Biomarcadores/líquido cefalorraquídeo , Electroforesis en Gel Bidimensional , Femenino , Estudios de Seguimiento , Humanos , Hidrocéfalo Normotenso/diagnóstico , Presión Intracraneal , Masculino , Selección de Paciente , Mapeo Peptídico , Proyectos Piloto , Proteoma , Espectrometría de Masa por Láser de Matriz Asistida de Ionización Desorción , Factores de Tiempo , Resultado del Tratamiento
14.
J Am Soc Mass Spectrom ; 20(1): 112-23, 2009 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-18849171

RESUMEN

A new analytical approach using matrix-assisted laser desorption/ionization mass spectrometry imaging (MALDI MSI) for the study of honeybee venom is shown. In vitro and in vivo models simulating the bee sting have been developed using live honeybees and, as the envenomation sites, pig ears and rat legs; MALDI MSI has been used to map, over time, the diffusion and distribution of three venom allergens (Api m 1, Api m 4, and Api m 6) and two venom toxins (apamine and mast cell degranulating peptide). In conjunction with other classical biochemical techniques and high resolution mass spectrometry (HRMS), structural data have been obtained that contribute to current understanding of honeybee venom composition. Initial data have also been obtained demonstrating the feasibility of mapping the organism's response to the sting. The opportunity to monitor venom diffusion and the organism's response at the same time might open new pathways for in vivo preclinical studies in designing and testing new venom immunotherapy (VIT).


Asunto(s)
Venenos de Abeja/análisis , Mordeduras y Picaduras de Insectos/metabolismo , Espectrometría de Masa por Láser de Matriz Asistida de Ionización Desorción/métodos , Alérgenos/análisis , Alérgenos/química , Animales , Antígenos de Plantas , Apamina/análisis , Apamina/química , Venenos de Abeja/química , Abejas , Oído , Extremidades , Mordeduras y Picaduras de Insectos/patología , Proteínas de Insectos/análisis , Proteínas de Insectos/química , Modelos Biológicos , Músculo Esquelético/química , Músculo Esquelético/patología , Péptidos/análisis , Péptidos/química , Fosfolipasas A/análisis , Fosfolipasas A/química , Ratas , Sensibilidad y Especificidad , Porcinos
15.
J Biol Inorg Chem ; 14(5): 761-70, 2009 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-19288144

RESUMEN

Electrospray ionisation mass spectrometry was used to analyse the reactions of metal compounds with mixtures of selected proteins. Three representative medicinally relevant compounds, cisplatin, transplatin and the organometallic ruthenium compound RAPTA-C, were reacted with a pool of three proteins, ubiquitin, cytochrome c and superoxide dismutase, and the reaction products were analysed using high-resolution mass spectrometry. Highly informative electrospray ionisation mass spectra were acquired following careful optimisation of the experimental conditions. The formation of metal-protein adducts was clearly observed for the three proteins. In addition, valuable information was obtained on the nature of the protein-bound metallofragments, on their distribution among the three different proteins and on the binding kinetics. The platinum compounds were less reactive and considerably less selective in protein binding than RAPTA-C, which showed a high affinity towards ubiquitin and cytochrome c, but not superoxide dismutase. In addition, competition studies between cisplatin and RAPTA-C showed that the two metallodrugs have affinities for the same amino acid residues on protein binding.


Asunto(s)
Cisplatino/metabolismo , Compuestos Organometálicos/metabolismo , Proteínas/metabolismo , Espectrometría de Masa por Ionización de Electrospray/métodos , Animales , Unión Competitiva , Bovinos , Cisplatino/química , Cimenos , Citocromos c/metabolismo , Caballos , Estructura Molecular , Compuestos Organometálicos/química , Unión Proteica , Relación Estructura-Actividad , Superóxido Dismutasa/metabolismo , Ubiquitina/metabolismo
16.
Epilepsia ; 50(12): 2658-62, 2009 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-19682026

RESUMEN

Advantages of dried blood spot include low invasiveness, ease and low cost of sample collection, transport, and storage. We used tandem mass spectrometry (LC-MS/MS) to determine phenobarbital levels on dried blood spot specimens and compared this methodology to commercially available particle enhanced turbidimetric inhibition immunoassay (PETINIA) in plasma/serum samples. The calibration curve in matrix using D(5)-phenobarbital as internal standard was linear in the phenobarbital concentration range of 1-100 mg/L (correlation coefficient 0.9996). The coefficients of variation in blood spots ranged 2.29-6.71% and the accuracy ranged 96.54-103.87%. There were no significant differences between the concentrations measured using PETINA and LC-MS/MS (both had similar precision and accuracy) however, LC-MS/MS allows at least 1.5 times higher throughput of phenobarbital analysis and additionally offers ease of sample collection which is particularly important for newborns or small infants.


Asunto(s)
Anticonvulsivantes/sangre , Análisis Químico de la Sangre/métodos , Cromatografía Liquida/métodos , Fenobarbital/sangre , Espectrometría de Masas en Tándem/métodos , Bioensayo , Análisis Químico de la Sangre/estadística & datos numéricos , Recolección de Muestras de Sangre/métodos , Calibración , Cromatografía Líquida de Alta Presión , Humanos , Inmunoensayo/métodos , Lactante , Recién Nacido , Persona de Mediana Edad , Nefelometría y Turbidimetría/métodos , Plasma/química , Sensibilidad y Especificidad , Suero/química , Espectrometría de Masa por Ionización de Electrospray
17.
J Chromatogr A ; 1180(1-2): 138-50, 2008 Feb 08.
Artículo en Inglés | MEDLINE | ID: mdl-18177880

RESUMEN

The qualitative and quantitative determination of components of mainstream and sidestream smoke has been performed by solid-phase microextraction-gas chromatography-mass spectrometry. Several brands and types of cigarettes sold in Italy were considered: normal, mild, light, extra light, some with filter and some without. Extraction of the analytes was performed by means of solid-phase microextraction (SPME) and the optimisation of the extraction procedure was performed by experimental design, taking into consideration type of fiber polymer, exposure temperature and time. Sixty-seven components of mainstream and sidestream smoke were identified. The quantified compounds (by means of deuterium-labelled isotopologues) were benzene, toluene, p-xylene, m-xylene, pyridine, o-xylene, limonene, naphthalene, phenol and nicotine. Finally, a comparison between the chemical profile of smoke from the different cigarettes was made.


Asunto(s)
Cromatografía de Gases y Espectrometría de Masas/métodos , Humo/análisis , Nicotiana
18.
J Pharm Biomed Anal ; 48(5): 1392-6, 2008 Dec 15.
Artículo en Inglés | MEDLINE | ID: mdl-18980824

RESUMEN

Topiramate (TPM) is a new antiepiletic drug with efficacy in several types of seizures. Therapeutic drug monitoring of TPM is essential for effective patient management. The aim of this study was to evaluate the use of dried blood spot (DBS) specimens to determinate the TPM levels during the treatment. Advantages of DBS include short collection time, low invasiveness, ease and low cost of sample collection, transport and storage. Performance comparison between this method and the commercially available fluorescence-polarization immunoassay (FPIA) was made. The analysis was performed in selected reaction monitoring (SRM) mode. The calibration curve in matrix using D(12)-topiramate was linear in the concentration range of 0.0166-1.66mg/L (0.5-50mg/L in DBS) of topiramate with correlation coefficient value of 0.9985. In the concentration range of 0.5-50mg/L, the coefficients of variation in DBS were in the range 2.13-11.85% and the accuracy ranged from 93.93% to 110.67%. There was no significant differences between the concentrations (ranging 0.5-50mg/L) measured both FPIA on venous samples and LC-MS/MS assay on simultaneous DBS samples. The sensitivity and specificity of tandem mass spectrometry allow now high throughput topiramate analysis (the improvement was three times in comparison with FPIA). This new assay has favourable characteristics being highly precise and accurate. FPIA also proved to be precise and accurate, but is not always suitable for the sample collection in neonates in whom obtaining larger blood samples is not convenient or possible.


Asunto(s)
Anticonvulsivantes/análisis , Recolección de Muestras de Sangre/métodos , Desecación/métodos , Fructosa/análogos & derivados , Espectrometría de Masas en Tándem/métodos , Anticonvulsivantes/química , Bioensayo/métodos , Calibración , Estudios de Casos y Controles , Cromatografía Liquida/métodos , Epilepsia/sangre , Polarización de Fluorescencia/métodos , Fructosa/análisis , Fructosa/química , Humanos , Inmunoensayo , Estructura Molecular , Estándares de Referencia , Sensibilidad y Especificidad , Factores de Tiempo , Topiramato
19.
Front Plant Sci ; 9: 1480, 2018.
Artículo en Inglés | MEDLINE | ID: mdl-30356724

RESUMEN

Arbuscular mycorrhizal (AM) fungi are very widespread, forming symbiotic associations with ∼80% of land plant species, including almost all crop plants. These fungi are considered of great interest for their use as biofertilizer in low-input and organic agriculture. In addition to an improvement in plant nutrition, AM fungi have been reported to enhance plant tolerance to important abiotic and biotic environmental conditions, especially to a reduced availability of resources. These features, to be exploited and applied in the field, require a thorough identification of mechanisms involved in nutrient transfer, metabolic pathways induced by single and multiple stresses, physiological and eco-physiological mechanisms resulting in improved tolerance. However, cooperation between host plants and AM fungi is often related to the specificity of symbiotic partners, the environmental conditions and the availability of resources. In this study, the impact of two AM fungal species (Funneliformis mosseae and Rhizophagus intraradices) on the water stress tolerance of a commercial tomato cultivar (San Marzano nano) has been evaluated in pots. Biometric and eco-physiological parameters have been recorded and gene expression analyses in tomato roots have been focused on plant and fungal genes involved in inorganic phosphate (Pi) uptake and transport. R. intraradices, which resulted to be more efficient than F. mosseae to improve physiological performances, was selected to assess the role of AM symbiosis on tomato plants subjected to combined stresses (moderate water stress and aphid infestation) in controlled conditions. A positive effect on the tomato indirect defense toward aphids in terms of enhanced attraction of their natural enemies was observed, in agreement with the characterization of volatile organic compound (VOC) released. In conclusion, our results offer new insights for understanding the molecular and physiological mechanisms involved in the tolerance toward water deficit as mediated by a specific AM fungus. Moreover, they open new perspectives for the exploitation of AM symbiosis to enhance crop tolerance to abiotic and biotic stresses in a scenario of global change.

20.
Org Lett ; 9(23): 4685-8, 2007 Nov 08.
Artículo en Inglés | MEDLINE | ID: mdl-17927190

RESUMEN

Acetalic substituents strategically located in a pyrrolic tripodal structure provide a new synthetic receptor endowed with unprecedented affinity for mannosides and the highest selectivity for beta-mannose ever reported for synthetic H-bonding receptors. Binding properties have been determined by NMR, ITC, and ESI-MS techniques, while affinities have been univocally assessed by the BC50(0) parameter, a general descriptor of binding affinity.


Asunto(s)
Manósidos/química , Pirroles/química , Estructura Molecular , Monosacáridos/química , Espectrometría de Masa por Ionización de Electrospray , Espectrometría de Masas en Tándem
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