RESUMEN
Very little is known about the influence of environmental radiation on living matter. In principle, important information can be acquired by analysing possible differences between parallel biological systems, one in a reference-radiation environment (RRE) and the other in a low-radiation environment (LRE). We took advantage of the unique opportunity represented by the cell culture facilities at the Gran Sasso National Laboratories of the Istituto Nazionale di Fisica Nucleare, where environment dose rate reduction factors in the underground (LRE), with respect to the external laboratory (RRE), are as follows: 10(3) for neutrons, 10(7) for directly ionizing cosmic rays and 10 for total γ-rays. Chinese hamster V79 cells were cultured for 10 months in both RRE and LRE. At the end of this period, all the cultures were kept in RRE for another 6 months. Changes in the activities of antioxidant enzymes (superoxide dismutase, SOD; catalase, CAT; glutathione peroxidase, GPX) and spontaneous mutation frequency at the hypoxanthine-guanine phosphoribosyl transferase (hprt) locus were investigated. The results obtained suggest that environmental radiation might act as a trigger of defence mechanisms in V79 cells, specifically those in reference conditions, showing a higher degree of defence against endogenous damage as compared to cells grown in a very low-radiation environment. Our findings corroborate the hypothesis that environmental radiation contributes to the development of defence mechanisms in today living organisms/systems.
Asunto(s)
Ambiente , Fibroblastos/efectos de la radiación , Animales , Antioxidantes/metabolismo , Línea Celular , Cricetulus , Relación Dosis-Respuesta en la Radiación , Fibroblastos/enzimología , Fibroblastos/metabolismo , Regulación de la Expresión Génica/efectos de la radiación , RadiometríaRESUMEN
BACKGROUND: The DNA-repair gene DNA-dependent kinase catalytic subunit (DNA-PKcs) favours or inhibits carcinogenesis, depending on the cancer type. Its role in human hepatocellular carcinoma (HCC) is unknown. METHODS: DNA-dependent protein kinase catalytic subunit, H2A histone family member X (H2AFX) and heat shock transcription factor-1 (HSF1) levels were assessed by immunohistochemistry and/or immunoblotting and qRT-PCR in a collection of human HCC. Rates of proliferation, apoptosis, microvessel density and genomic instability were also determined. Heat shock factor-1 cDNA or DNA-PKcs-specific siRNA were used to explore the role of both genes in HCC. Activator protein 1 (AP-1) binding to DNA-PKcs promoter was evaluated by chromatin immunoprecipitation. Kaplan-Meier curves and multivariate Cox model were used to study the impact on clinical outcome. RESULTS: Total and phosphorylated DNA-PKcs and H2AFX were upregulated in HCC. Activated DNA-PKcs positively correlated with HCC proliferation, genomic instability and microvessel density, and negatively with apoptosis and patient's survival. Proliferation decline and massive apoptosis followed DNA-PKcs silencing in HCC cell lines. Total and phosphorylated HSF1 protein, mRNA and activity were upregulated in HCC. Mechanistically, we demonstrated that HSF1 induces DNA-PKcs upregulation through the activation of the MAPK/JNK/AP-1 axis. CONCLUSION: DNA-dependent protein kinase catalytic subunit transduces HSF1 effects in HCC cells, and might represent a novel target and prognostic factor in human HCC.
Asunto(s)
Biomarcadores de Tumor/genética , Carcinogénesis/genética , Carcinoma Hepatocelular/patología , Proteína Quinasa Activada por ADN/genética , Neoplasias Hepáticas/patología , Proteínas Nucleares/genética , Carcinoma Hepatocelular/genética , Línea Celular Tumoral , Proliferación Celular , Supervivencia Celular/genética , Proteínas de Unión al ADN/fisiología , Regulación Enzimológica de la Expresión Génica , Regulación Neoplásica de la Expresión Génica , Factores de Transcripción del Choque Térmico , Células Hep G2 , Humanos , Neoplasias Hepáticas/genética , Valor Predictivo de las Pruebas , Pronóstico , Factores de Transcripción/fisiologíaRESUMEN
UNLABELLED: We report a case of a parathyroid adenoma during a long term lithium treatment without therapeutic overdose. CASE REPORT: A 73-years-old woman presented a demonstrative biological syndrome with hypercalcemia, elevated parathormone, normal urinary cyclic AMP, normocalciuria. CONCLUSION: This lithium induced hyperparathyroidism differs from the classic primary hyperparathyroidism with parthyroid adenoma where urinary cyclic AMP excretion is elevated and where there is hypercalciuria. Lithium is blocking the negative feedback of calcium on parathormone secretion and stimulates the growth of parathyroid adenoma. Treatment is surgical and consists in adenoma ablation. Calcemia follow up is indicated in patients with long term lithium therapy
Asunto(s)
Adenoma/inducido químicamente , Antipsicóticos/efectos adversos , Compuestos de Litio/efectos adversos , Neoplasias de las Paratiroides/inducido químicamente , Adenoma/diagnóstico , Adenoma/diagnóstico por imagen , Adenoma/cirugía , Anciano , Trastorno Bipolar/tratamiento farmacológico , Femenino , Humanos , Hipercalcemia/complicaciones , Hiperparatiroidismo/inducido químicamente , Hiperparatiroidismo/complicaciones , Compuestos de Litio/administración & dosificación , Neoplasias de las Paratiroides/diagnóstico por imagen , Neoplasias de las Paratiroides/cirugía , Cintigrafía , Factores de Tiempo , Tomografía Computarizada por Rayos XRESUMEN
BACKGROUND AND AIMS: Previous studies indicate unrestrained cell cycle progression in liver lesions from hepatocarcinogenesis-susceptible Fisher 344 (F344) rats and a block of G(1)-S transition in corresponding lesions from resistant Brown Norway (BN) rats. Here, the role of the Forkhead box M1B (FOXM1) gene during hepatocarcinogenesis in both rat models and human hepatocellular carcinoma (HCC) was assessed. METHODS AND RESULTS: Levels of FOXM1 and its targets were determined by immunoprecipitation and real-time PCR analyses in rat and human samples. FOXM1 function was investigated by either FOXM1 silencing or overexpression in human HCC cell lines. Activation of FOXM1 and its targets (Aurora Kinose A, Cdc2, cyclin B1, Nek2) occurred earlier and was most pronounced in liver lesions from F344 than BN rats, leading to the highest number of Cdc2-cyclin B1 complexes (implying the highest G(2)-M transition) in F344 rats. In human HCC, the level of FOXM1 progressively increased from surrounding non-tumorous livers to HCC, reaching the highest levels in tumours with poorer prognosis (as defined by patients' length of survival). Furthermore, expression levels of FOXM1 directly correlated with the proliferation index, genomic instability rate and microvessel density, and inversely with apoptosis. FOXM1 upregulation was due to extracellular signal-regulated kinase (ERK) and glioblastoma-associated oncogene 1 (GLI1) combined activity, and its overexpression resulted in increased proliferation and angiogenesis and reduced apoptosis in human HCC cell lines. Conversely, FOXM1 suppression led to decreased ERK activity, reduced proliferation and angiogenesis, and massive apoptosis of human HCC cell lines. CONCLUSIONS: FOXM1 upregulation is associated with the acquisition of a susceptible phenotype in rats and influences human HCC development and prognosis.
Asunto(s)
Carcinoma Hepatocelular/genética , Quinasas MAP Reguladas por Señal Extracelular/genética , Factores de Transcripción Forkhead/genética , Neoplasias Hepáticas/genética , Animales , Carcinoma Hepatocelular/patología , Proliferación Celular , Transformación Celular Neoplásica/genética , Proteína Forkhead Box M1 , Factores de Transcripción Forkhead/fisiología , Regulación Neoplásica de la Expresión Génica/genética , Genes cdc , Predisposición Genética a la Enfermedad/genética , Hígado/patología , Neoplasias Hepáticas/patología , Neovascularización Patológica/etiología , Ratas , Ratas Endogámicas F344 , Activación Transcripcional , Regulación hacia ArribaRESUMEN
Fracture of the nose is a frequent injury. Careful management is necessary to avoid not only cosmetic but also functional sequels. Therapeutic modalities are simple and can easily be carried out under local anesthesia.
Asunto(s)
Fracturas Cerradas/diagnóstico , Fracturas Cerradas/cirugía , Nariz/lesiones , Nariz/cirugía , Anestesia Local , HumanosRESUMEN
Foreign bodies in the nasal fossa are frequent and generally occur in children. In developing countries, access to an ENT specialist can be difficult or impossible. The authors describe several extraction techniques with special emphasis on those best suited to areas with limited access to specialist facilities. Using illustrations, a step-by-step description of the so-called "hook" technique is given. This simple technique allows successful removal of a foreign body from the nasal fossa in almost all cases.
Asunto(s)
Cuerpos Extraños/cirugía , Otolaringología/instrumentación , Instrumentos Quirúrgicos , Países en Desarrollo , Diseño de Equipo , Humanos , Cavidad NasalRESUMEN
OBJECTIVES: The main purpose of the article is to raise awareness among all the involved stakeholders about the risks and legal implications connected to the development and use of modern telemedicine systems. Particular focus is given to the class of "active" telemedicine systems, that imply a real-world, non-mediated, interaction with the final user. A secondary objective is to give an overview of the European legal framework that applies to these systems, in the effort to avoid defensive medicine practices and fears, which might be a barrier to their broader adoption. METHODS: We leverage on the experience gained during two international telemedicine projects, namely MobiGuide (pilot studies conducted in Spain and Italy) and AP@home (clinical trials enrolled patients in Italy, France, the Netherlands, United Kingdom, Austria and Germany), whose development our group has significantly contributed to in the last 4 years, to create a map of the potential criticalities of active telemedicine systems and comment upon the legal framework that applies to them. Two workshops have been organized in December 2015 and March 2016 where the topic has been discussed in round tables with system developers, researchers, physicians, nurses, legal experts, healthcare economists and administrators. RESULTS: We identified 8 features that generate relevant risks from our example use cases. These features generalize to a broad set of telemedicine applications, and suggest insights on possible risk mitigation strategies. We also discuss the relevant European legal framework that regulate this class of systems, providing pointers to specific norms and highlighting possible liability profiles for involved stakeholders. CONCLUSIONS: Patients are more and more willing to adopt telemedicine systems to improve home care and day-by-day self-management. An essential step towards a broader adoption of these systems consists in increasing their compliance with existing regulations and better defining responsibilities for all the involved stakeholders.
Asunto(s)
Atención a la Salud , Responsabilidad Legal , Seguridad del Paciente , Gestión de Riesgos , Telemedicina/legislación & jurisprudencia , Telemedicina/normas , Europa (Continente) , Humanos , Participación de los InteresadosRESUMEN
Incubation of Sendai virus with pyridoxal 5'-phosphate (PLP) causes inhibition of hemolytic activity, a slight reduction of hemagglutinating activity, and an increase in neuraminidase activity. The effects on hemagglutination and neuraminidase are prevented by the presence in the incubation mixture of sialyl lactose, a substrate of hemagglutinin-neuraminidase. Incubation with PLP of the water-soluble enzymatic domain of the neuraminidase has no effect on enzymatic activity, while the allosteric inhibition (Dallocchio et al. (1991) Biochem. Int. 25, 663-668) disappears. Both virus-bound and solubilized neuraminidase are selectively modified by PLP at the lysine-553. Our data suggest that PLP inactivates a previously undetected inhibitory site on the viral neuraminidase, and that a physiological effector is present on the viral envelope.
Asunto(s)
Proteína HN/metabolismo , Virus de la Parainfluenza 1 Humana/enzimología , Fosfato de Piridoxal/metabolismo , Regulación Alostérica , Aminoácidos/análisis , Cromatografía Líquida de Alta Presión , Proteína HN/química , Hemaglutinación por Virus , Humanos , Lisina/metabolismo , Especificidad por SustratoRESUMEN
A highly purified amylase protein inhibitor from the kernels of hexaplois wheat, designated 0.19 according to its gel electrophoretic mobility, has been characterized according to its circular dichroism spectra determined at different pH values and in the presence or absence of dissociating and reducing agents. The 0.19 albumin has also been characterized according to the specificity with which it inhibits 21 alpha-amylases from different origins and according to its sensitivity to a number of chemical and enzymatic treatments of its inhibitory action on human saliva and Tenebrio molitor L. larval midgut alpha-amylases. Inhibitory activity of 0.19 toward human saliva amylase significantly increased when the inhibitor was incubated with the enzyme before the addition of starch, but it was not affected by the preincubation of 0.19 with starch. Maltose reversed the inhibition of human saliva by 0.19 and showed some inhibitory activity toward the enzyme. However, maltose concentrations that only slightly affected amylase activity were very effective in restoring the amylase activity inhibited by 0.19. The inhibitory action of 0.19 on human saliva and T. molitor L. amylases were equally resistant to trypsin and thermal treatments, but 0.19 was readily inactivated by incubation with pepsin or by reduction of disulfide bonds. The inhibition of the mammalian amylase by 0.19 was adversely affected by a treatment with CNBr (1:100 ratio of methionine residues to CNBr) whereas the inhibition of the insect amylase was not. As shown by circular dichroism measurements in the far ultraviolet, 0.19 is a protein with about 50% of ordered structure. Significant and largely reversible changes have been observed in the aromatic CD spectrum of 0.19 at alkaline pH values or in the presence of sodium dodecyl sulfate. These changes, which were associated with a partial loss of inhibitory activity, indicate that ionizable tyrosine groups contribute significantly to the ellipticity bands of 0.19 in the near ultraviolet.
Asunto(s)
Amilasas/antagonistas & inhibidores , Proteínas de Plantas , Semillas/análisis , Dicroismo Circular , Bromuro de Cianógeno , Humanos , Cinética , Proteínas de Plantas/farmacología , Conformación Proteica , Espectrofotometría UltravioletaRESUMEN
The topology of the interaction of cholera toxin with ganglioside and detergent micelles was studied with the technique of hydrophobic photolabelling. Cholera toxin alpha and gamma polypeptide chains appear to penetrate into the hydrophobic core of ganglioside micelles. Micelles of SDS cause the labelling also of the beta polypeptide chains, while Triton X-100 micelles have little ability to mediate the labelling of the toxin. The specific reduction of the alpha-gamma disulfide bond allows the penetration of the alpha polypeptide chain into Triton X-100 micelles, but does not affect the interaction of cholera toxin with either ganglioside or SDS micelles. Thus, ganglioside micelles appear to cause a conformational change of the native toxin, such as to induce the penetration of the alpha chain into the micelle hydrophobic core.
Asunto(s)
Toxina del Cólera , Coloides , Gangliósidos , Micelas , Detergentes , Disulfuros , Cinética , Oxidación-Reducción , Unión Proteica , TemperaturaRESUMEN
The heterogeneity of Vibrio cholerae toxin, obtained from culture filtrates in homogeneous form by gel filtration and preparative disc gel electrophoresis has been studied. By means of disc electrophoresis on polyacrylamide gel cholera toxin was separated into three forms designated I (5%), II (15%) and III (80%). The toxic activity, amino acid content and molecular weight of the three forms were similar. The difference so far observed between the various electrophoretic fractions is a difference in net charge. Incubation of either cholera toxin II or cholera toxin III at relatively high pH leads to the formation of the more acidic forms. These forms, generated in vitro by deamidation of asparagine and/or glutamine residues, are indistinguishable from the toxins of similar electrophoretic mobilities isolated from crude culture filtrates.
Asunto(s)
Toxinas Bacterianas , Toxinas Bacterianas/aislamiento & purificación , Dicroismo Circular , Peso Molecular , Fragmentos de Péptidos/análisis , Conformación Proteica , Vibrio choleraeRESUMEN
The hydrophobic surfaces presumably involved in the membrane interaction of pertussis toxin have been mapped by a new detergent-binding assay. This is based on the interdispersion among detergent micelles of trace amounts of radioactive photoreactive phospholipid analogues, able to cross-link to the protein thereby labelling its detergent-binding domains. The assay has proven to be very sensitive. Subunits B1, B2 and B3 of pertussis toxin were found to interact with the lipid micelles suggesting that they may be involved in the membrane penetration step of the intoxication process.
Asunto(s)
Toxina del Pertussis , Fosfolípidos/análisis , Factores de Virulencia de Bordetella/análisis , Detergentes , Interacciones Farmacológicas , Lisofosfatidilcolinas/análisis , Micelas , Peso Molecular , Octoxinol , Polietilenglicoles , Ensayo de Unión Radioligante/métodos , Dodecil Sulfato de SodioRESUMEN
Association of the ribosome-inactivating proteins (RIPs): pokeweed antiviral protein (PAP), gelonin, Momordica charantia inhibitor (MCI), with reconstituted Sendai virus envelopes (RSVE) was obtained without detectable loss of activities either of RIPs or of viral envelope glycoproteins. RIPs are inactive towards intact cells, but, once encapsulated in RSVE, they become cytotoxic. The concentration of RSVE-associated PAP, which causes 50% inhibition of protein synthesis by Friend erythroleukemic cells, is 0.5 ng/ml. Substances capable to inhibit the viral activities block the acquired cytotoxicity of RIPs associated to RSVE.
Asunto(s)
Citotoxinas/metabolismo , Virus de la Parainfluenza 1 Humana/metabolismo , Ribosomas/efectos de los fármacos , Animales , Células Cultivadas , Cricetinae , Cricetulus , Virus de la Leucemia Murina de Friend , Ratones , Proteínas de Plantas/farmacología , Proteínas Inactivadoras de Ribosomas Tipo 1 , Proteínas Virales/farmacologíaRESUMEN
The domain structure of hemagglutinin-neuraminidase from Sendai virus (cHN) was investigated by studying the thermal stability in the 20-100 degrees C range. Differential scanning calorimetry evidences two conformational transitions. The first transition is apparently a reversible two-state process, with Tm 48.3 degrees C, and is shifted to 50.1 degrees C in the presence of the substrate analogue 2,3-dehydro-2-deoxy-N-acetyl neuraminic acid, meaning that the substrate binding domain is involved in the transition. The second transition, with apparent Tm 53.2 degrees C, is accompanied by irreversible loss of enzymatic activity of the protein, and the presence of the substrate analogue does not affect the Tm. The data indicate that cHN is composed of two independent folding domains, and that only one domain is involved in the binding of the substrate. Our results suggest that the paramyxovirus neuraminidases have the folding properties of a two-domain protein.
Asunto(s)
Proteína HN/química , Proteína HN/metabolismo , Ácido N-Acetilneuramínico/análogos & derivados , Pliegue de Proteína , Respirovirus/enzimología , Temperatura , Calorimetría , Estabilidad de Enzimas/fisiología , Fluorescencia , Ácido N-Acetilneuramínico/química , Péptido Hidrolasas/metabolismo , Desnaturalización Proteica/fisiología , Estructura Terciaria de Proteína/fisiología , TermodinámicaRESUMEN
Incubation of trifluoperazine, a local anaesthetic, at concentrations higher than the cmc with Sendai virus particles produces the selective solubilization of the haemagglutinin neuraminidase (HN) and matrix (M) proteins. This phenomenon involves aggregation of the Sendai virions and therefore the separation of HN and M from the rest of the particle can be performed by bench centrifugation. The supernatant contains the HN and M proteins and HN, once inserted into liposomes, elicits its own biological activities. Therefore, the method seems suitable for purifying large amounts of HN.
Asunto(s)
Hemaglutininas Virales/aislamiento & purificación , Virus de la Parainfluenza 1 Humana/análisis , Proteínas de la Matriz Viral/aislamiento & purificación , Virión/análisis , Animales , Embrión de Pollo , Detergentes , Neuraminidasa/aislamiento & purificación , Virus de la Parainfluenza 1 Humana/enzimología , Solubilidad , Trifluoperazina , Virión/enzimologíaRESUMEN
The interaction of diphtheria toxin and its enzymatically deficient mutants crm 176 and crm 197 with liposomes has been studied by turbidity measurement and hydrophobic photolabelling with photoactivatable phosphatidylcholines. Diphtheria toxin and crm 176 at neutral pH bind to the surface of lipid bilayers while crm 197 also appears to interact with the fatty acid chains of phospholipids. All proteins undergo a change in conformation over the same range of acidic pH and become able to insert in the lipid bilayer. The tighter lipid interaction of crm 197 may account for its higher cell association constant. The possibility is discussed that the binding of diphtheria toxin to cells is mediated by both a protein receptor and an interaction with the head group of phospholipids.
Asunto(s)
Toxina Diftérica/genética , Metabolismo de los Lípidos , Receptores de Superficie Celular , Receptores Colinérgicos/metabolismo , Toxina Diftérica/metabolismo , Factor de Crecimiento Similar a EGF de Unión a Heparina , Concentración de Iones de Hidrógeno , Péptidos y Proteínas de Señalización Intercelular , Liposomas , Nefelometría y Turbidimetría , Fosfolípidos/metabolismoRESUMEN
The Sendai virus fuses with host cell membranes in a pH-independent manner through an unknown mechanism. Here we report that mild trypsin pre-treatments of Sendai virions, for example 15 min at 4 degrees C, give Sendai virions the ability to fuse at a rate up to 10-fold higher than control. By using human erythrocytes as host cell membranes, viral fusion was assessed by hemolysis as well as fluorescence dequenching of octadecyl rhodamine B chloride. The mild protease treatment strikingly shortens the lag time taken by the virus to start the fusion process. Similar data were obtained on reconstituted Sendai virus envelope. Among proteases, tested as fusion enhancer, trypsin is more effective than either endoproteinase Lys-C, chymotrypsin, or endoproteinase Arg-C. After removal of trypsin from treated virions the fusion rate enhancement remains for hours at room temperature. The lack of protease specificity, together with the impossibility to detect any new N-terminal products, suggests that only a small percentage of viral envelope components are cleaved, still a large enough number to set the envelope in a ready-to-fuse state.
Asunto(s)
Endopeptidasas/farmacología , Respirovirus/metabolismo , Quimotripsina/metabolismo , Eritrocitos/metabolismo , Colorantes Fluorescentes/metabolismo , Humanos , Cinética , Fusión de Membrana/fisiología , Metaloendopeptidasas/metabolismo , Rodaminas/metabolismo , Serina Endopeptidasas/metabolismo , Tripsina/metabolismo , Proteínas del Envoltorio Viral/metabolismoRESUMEN
BACKGROUND: The paraneoplastic syndromes can involve multiple areas of the central nervous system and result in a variety of neurological symptoms. To our knowledge, severe, rapidly progressive, and drug-resistant ballistic-choreic movements have not been previously described as the presenting feature of renal cell carcinoma. PATIENT AND METHODS: A previously healthy 55-year-old man developed limb ballismus and involuntary choreic movements of his face over several weeks. Extensive laboratory, diagnostic, and radiographic studies failed to reveal a cause, until an abnormality on a chest x-ray film prompted a search for a primary neoplasm and a final diagnosis of renal cell carcinoma. High doses of medications traditionally used to treat choreic disorders had no effect on the abnormal movements. A biopsy specimen of the basal ganglia showed focal encephalitic changes but no malignant neoplasm. CONCLUSIONS: Whereas prior cases of paraneoplastic syndromes with chorea have been reported in other forms of cancer, our case was significant because, to our knowledge, renal cell carcinoma has not been previously reported in association with this syndrome. Furthermore, the chorea was categorically resistant to pharmacological treatment, and the movement disorder was the initial and only focal neurological feature of the primary illness.
Asunto(s)
Carcinoma de Células Renales/complicaciones , Carcinoma de Células Renales/diagnóstico , Discinesias/etiología , Neoplasias Renales/complicaciones , Neoplasias Renales/diagnóstico , Síndromes Paraneoplásicos/diagnóstico , Ganglios Basales/patología , Biopsia , Carcinoma de Células Renales/patología , Corea/etiología , Diagnóstico Diferencial , Humanos , Neoplasias Renales/patología , Masculino , Persona de Mediana Edad , Síndromes Paraneoplásicos/patologíaRESUMEN
Several immunoadjuvant systems have been proposed to enhance mucosal immune responses of orally administered purified antigens. Cholera toxin (CT) or its B subunit (CTB) have been found to promote immune responses to antigens when they are co-administered via mucosal routes. Oral administration of antigens incorporated into liposomes has also been shown to result in enhanced mucosal immune responses. Here, we describe the covalent coupling of CT and CTB to small unilamellar liposomes for targeting these vesicles to Peyer's patch M cells, following their oral administration. Conjugation was done by means of a thioether bond using succinimidyl(4-N-maleimidomethyl)cyclohexane-1-carboxylate to modify the dipalmitoylphosphatidyl-ethanolamine constituent of liposomes and N-succinimidyl-3-(2-pyridyldithio)propionate to thiolate CT or CTB. The biological activity of CT or CTB bound to liposomes was confirmed by a hemagglutination assay using GM1-enriched human erythrocytes. Furthermore, oral administration of CT-conjugated liposomes to rats resulted in the induction of serum IgG and salivary IgA anti-CT responses. CT-conjugated liposomes may prove to be a useful system for targeted delivery and immunoenhancement of weakly immunogenic antigens.
Asunto(s)
Adyuvantes Inmunológicos/química , Antígenos/administración & dosificación , Toxina del Cólera/química , Liposomas/química , Animales , Anticuerpos Antibacterianos/inmunología , Toxina del Cólera/administración & dosificación , Toxina del Cólera/inmunología , Inmunoglobulina A Secretora/inmunología , Inmunoglobulina G/inmunología , Fosfatidiletanolaminas/química , Ratas , Ratas Endogámicas F344RESUMEN
The glial C6BU-1 cell line, loaded with acetylcholine can release this neurotransmitter. This study was aimed at determining whether disruption of the Golgi-vesicular traffic by brefeldin A would change the acetylcholine release from these cells and affect proteins involved in transmitter release like the 15 kDa proteolipid, common to V-ATPase and mediatophore. Cells were treated for 24 or 36 h with brefeldin A (35.7 microM). The observed changes in cell morphology were typical for brefeldin A treated cells in which protein membrane supply has been stopped. Inhibition of membrane protein supply was confirmed in the present work. Moreover, the 15 kDa proteolipid also decayed to a very low level in the cell membrane fraction. The release of acetylcholine evoked by a calcium challenge and a calcium ionophore, or by electrical pulses decreased markedly. The life time of the release mechanism was of the order of 36 h and half decayed in 24 h. In addition, the electrically evoked release became much shorter. Considering that C6BU-1 cells are able to release large amounts of ACh and their membranes contain a sizeable amount of the 15 kDa proteolipid, these results suggest that this proteolipid may be one of the proteins forming the membrane complex responsible for transmitter release, at least in these cells.