RESUMEN
Single chain Fv (scFv) antibodies (generated by phage display technology, molecules representing new and efficient tools in the research and diagnostics of infectious diseases) against the capsid protein (p25) of Maedi-Visna virus were selected. Several clones of p25 specific scFv antibodies were identified; one of them was expressed as a soluble scFv molecule, purified by immobilized metal-affinity chromatography and further characterized by sequencing and determination of the kinetic equilibrium association constant. Sequence analysis showed that the rearranged VL and VH domains of the analyzed scFv clone used sequences from the VL3 family (germline DPL16/VL3.1) and VH1 family (germline VH20), respectively. The kinetic equilibrium association constant was determined as KA = 1.12 +/- 0.52 L/mumol.
Asunto(s)
Proteínas de la Cápside/inmunología , Cadenas Pesadas de Inmunoglobulina/inmunología , Cadenas Ligeras de Inmunoglobulina/inmunología , Región Variable de Inmunoglobulina/inmunología , Virus Visna-Maedi/inmunología , Secuencia de Aminoácidos , Animales , Secuencia de Bases , Proteínas de la Cápside/genética , Fragmentos de Inmunoglobulinas/genética , Fragmentos de Inmunoglobulinas/inmunología , Fragmentos de Inmunoglobulinas/aislamiento & purificación , Cadenas Pesadas de Inmunoglobulina/genética , Cadenas Ligeras de Inmunoglobulina/genética , Región Variable de Inmunoglobulina/genética , Región Variable de Inmunoglobulina/aislamiento & purificación , Cinética , Datos de Secuencia Molecular , Biblioteca de Péptidos , Proteínas Recombinantes/genética , Proteínas Recombinantes/inmunología , Proteínas Recombinantes/aislamiento & purificación , Análisis de Secuencia de ADN , Virus Visna-Maedi/genéticaRESUMEN
The development of piezoelectric immunosensors for human serum albumin (HSA) is reported. The piezoelectric crystals were modified either with monoclonal antibody AL-01 (direct assay) or with HSA (competitive assay). Measurements were carried out in the flow-through mode. Affinity interaction between albumin and the antibody was characterised. With immobilised antibody and HSA in solution, the kinetic association rate constant k(a) was 18 100 l mol(-1) s(-1) and the dissociation constant k(d) was 0.00369 s(-1). For the opposite arrangement (immobilised HSA), a slower dissociation was observed, k(d) was 0.00085 s(-1). A competitive assay for HSA was developed with working range of 1-5 mug ml(-1) and a total time for one analysis equal to 17 min. Samples of urine were analysed after tenfold dilution. The developed system was successfully evaluated on real samples from diabetic patients and the obtained results correlated well with the standard reflectometric assay of proteins in urine.
RESUMEN
The goal of this study was to determine subsequent malignancy on a discrete group of precancerous laryngeal lesions, and to assess the mortality. In a series of 227 patients, average age 51.8 years (ranging from 13 to 80 years). The changes were followed-up for 12.3 years (minimum of 5 years and maximum of 40 years). 58% are living without any sign of premalignant laryngeal mucosal disease, 13% with controlled precancer, and 3% in remission after surgery for carcinoma. 11% died (9% due to cause unrelated to the cancer) and 15% were lost for follow-up. 17% of the group with mucosal hyper- or metaplasia progressed to mild dysplasia, but none progressed to carcinoma. Reinke's oedema recurred in 4%, no malignancy was observed. Carcinoma developed in 16% of laryngeal papilloma (8% in situ and 8% invasive). 15% of mild dysplasia progressed in severity, but none transformed to malignancy. Moderate dysplasia progressed to severe dysplasia in 12%, carcinoma in situ in 4%. Of cases with severe dysplasia 13% developed in situ carcinoma while 43% progressed to invasive cancer. In the whole series progression to severe grade was seen in 7.1%, and malignant transformation in 4.4%. Three patients (1.3%) died due to subsequent carcinoma. Our results agree with some authors; but the majority of them reports higher incidence of malignant transformation. Invasive carcinoma was diagnosed in the follow-up in seven patients. Those represent only 3% of all laryngeal carcinomas diagnosed in our department in the same period of time. Based on the data we have evaluated the intensity of follow-up in patients with hyperplasia, metaplasia, keratosis and mild dysplasia.
Asunto(s)
Transformación Celular Neoplásica , Neoplasias Laríngeas/mortalidad , Neoplasias Laríngeas/patología , Lesiones Precancerosas/mortalidad , Lesiones Precancerosas/patología , Adolescente , Adulto , Anciano , Anciano de 80 o más Años , Supervivencia sin Enfermedad , Femenino , Estudios de Seguimiento , Humanos , Neoplasias Laríngeas/clasificación , Neoplasias Laríngeas/cirugía , Masculino , Microcirugia , Persona de Mediana Edad , Lesiones Precancerosas/clasificación , Lesiones Precancerosas/cirugía , Inducción de RemisiónRESUMEN
During the 15-year period from 1978 to 1993 we treated 17 cannulated patients suffering from dyspnea caused by bilateral recurrent laryngeal nerve palsies. The causes of palsy were struma surgery (13 cases), prolonged intubation (3 cases) and unknown (1 case). All cases were operated on by the classical endolaryngeal arytenoidectomy technique developed by Kleinsasser. All patients were able to be decanulated after arytenoidectomy: 14 cases within 1 month of surgery and 3 delayed. Four patients required replacements of their tracheotomies during the follow-up period, but this proved to be only temporary in 3 cases. After excisions of polypoid tissues or enlargement of an ipsilateral partial cordectomy, definitive closure of the tracheostome was possible. Among the retracheotomized patients, only 1 patient with multilocular stenoses has remained under treatment for the past 2 years. In all, 16 of 17 patients (93%) have remained permanently decannulated.