RESUMEN
Porcine brucellosis is one of the most important zoonoses in this country. Currently, there is no control program for porcine brucellosis in Argentina and the epidemiological situation is still unknown. The purpose of our study was to detect anti-Brucella spp. antibodies in swine in the southwest of the Buenos Aires province and the east of the La Pampa province. Blood samples were obtained when animals were slaughtered. The presence of anti-brucella antibodies was studied by the buffered plate agglutination test (BPA), the tube agglutination test (SAT), the 2-mercaptoethanol (2-ME) agglutination test and indirect ELISA tests, using the cytosolic fraction from Brucella abortus S19 (CYT), and lipopolysaccharide (LPS)-free cytosolic proteins (CP). Out of a total of 325 samples analyzed, 17.8% reacted positively to BPA, 13.8% to SAT, 8.0% to 2-ME, 21.0% to ELISA-CYT and 10.0% to ELISA-CP. These results agree with the few data available in our country and suggest that brucellosis screening should be extended to other regions.
Asunto(s)
Pruebas de Aglutinación , Anticuerpos Antibacterianos/aislamiento & purificación , Brucella/inmunología , Brucelosis/veterinaria , Enfermedades de los Porcinos/microbiología , Animales , Argentina/epidemiología , Brucelosis/diagnóstico , Brucelosis/epidemiología , Ensayo de Inmunoadsorción Enzimática , Porcinos , Enfermedades de los Porcinos/diagnóstico , Enfermedades de los Porcinos/epidemiologíaRESUMEN
We analyzed the influence of presenilins on the genetic cascades that control neuronal differentiation in Xenopus embryos. Resembling sonic hedgehog (shh) overexpression, presenilin mRNA injection reduced the number of N-tubulin+ primary neurons and modulated Gli3 and Zic2 according to their roles in activating and repressing primary neurogenesis, respectively. Presenilin increased shh expression within its normal domain, mainly in the floor plate, whereas an antisense X-presenilin-alpha morpholino oligonucleotide reduced shh expression. Both shh and presenilin promoted cell proliferation and apoptosis, but the effects of shh were widely distributed, while those resulting from presenilin injection coincided with the range of shh signaling. We suggest that presenilin may modulate primary neurogenesis, proliferation, and apoptosis in the neural plate, through the enhancement of shh signaling.
Asunto(s)
Proteínas de la Membrana/genética , Proteínas del Tejido Nervioso , Neuronas/citología , Proteínas Represoras , Transactivadores/genética , Proteínas de Xenopus , Xenopus laevis/embriología , Secretasas de la Proteína Precursora del Amiloide , Animales , Apoptosis , Ácido Aspártico Endopeptidasas , Diferenciación Celular , División Celular , Sistema Nervioso Central/embriología , Proteínas de Unión al ADN/genética , Regulación hacia Abajo , Embrión no Mamífero/citología , Embrión no Mamífero/metabolismo , Endopeptidasas/metabolismo , Regulación del Desarrollo de la Expresión Génica , Proteínas Hedgehog , Hibridación in Situ , Factores de Transcripción de Tipo Kruppel , Proteínas de la Membrana/fisiología , Mutagénesis Sitio-Dirigida , Oligonucleótidos Antisentido , Presenilina-1 , ARN Mensajero/genética , ARN Mensajero/metabolismo , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Transducción de Señal , Transactivadores/fisiología , Factores de Transcripción/genética , Tretinoina/farmacología , Tubulina (Proteína)/genética , Tubulina (Proteína)/metabolismo , Xenopus laevis/genética , Xenopus laevis/metabolismo , Proteína Gli3 con Dedos de ZincRESUMEN
Alzheimer's disease (AD) is characterized by the massive deposition in the brain of the 40-42-residue amyloid beta protein (A(beta)). While A(beta)1-40 predominates in the vascular system, A(beta)1-42 is the major component of the senile plaques in the neuropil. The concentration of both A(beta) species required to form amyloid fibrils in vitro is micromolar, yet soluble A(betas) found in normal and AD brains are in the low nanomolar range. It has been recently proposed that the levels of A(beta) sufficient to trigger amyloidogenesis may be reached intracellularly. To study the internalization and intracellular accumulation of the major isoforms of A(beta), we used THP-1 and IMR-32 neuroblastoma cells as models of human monocytic and/or macrophagic and neuronal lineages, respectively. We tested whether these cells were able to internalize and accumulate 125I-A(beta)1-40 and 125I-A(beta)1-42 differentially when offered at nanomolar concentrations and free of large aggregates, conditions that mimic a prefibrillar stage of A(beta) in AD brain. Our results showed that THP-1 monocytic cells internalized at least 10 times more 125I-A(betas) than IMR-32 neuroblastoma cells, either isolated or in a coculture system. Moreover, 125I-A(beta)1-42 presented a higher adsorption, internalization, and accumulation of undigested peptide inside cells, as opposed to 125I-A(beta)1-40. These results support that A(beta)1-42, the major pathogenic form in AD, may reach supersaturation and generate competent nuclei for amyloid fibril formation intracellularly. In light of the recently reported strong neurotoxicity of soluble, nonfibrillar A(beta)1-42, we propose that intracellular amyloidogenesis in microglia is a protective mechanism that may delay neurodegeneration at early stages of the disease.
Asunto(s)
Péptidos beta-Amiloides/metabolismo , Monocitos/metabolismo , Neuroblastoma/metabolismo , Fragmentos de Péptidos/metabolismo , Adsorción , Enfermedad de Alzheimer/metabolismo , Neuropatías Amiloides/metabolismo , Línea Celular , Técnicas de Cocultivo , Humanos , Radioisótopos de Yodo/análisis , Isoformas de Proteínas , Células Tumorales CultivadasRESUMEN
Microglial cell involvement in Alzheimer's disease has been related to amyloid beta (A beta) internalization, the release of inflammatory cytokines and the development of neuritic plaques. The human monocyte/macrophage THP-1 cell line has been widely used as a model of human microglial cells. We used THP-1 cells to study the adsorption, internalization and resistance to degradation of A beta1-40 and A beta1-42 isoforms offered at nanomolar concentrations and free of large aggregates, conditions that may mimic a pre-fibrillar stage of A beta in the brain. Under these conditions, A betas did not induce THP-1 activation, as assessed by interleukin-1beta expression. A beta1-42 showed a preferential adsorption and intracellular accumulation as compared to A beta1-40, supporting that competent nuclei for A beta1-42 ordered aggregation may be formed inside microglial cells. In light of the possible neurotoxicity of soluble A beta1-42, we propose that amyloid formation within brain phagocytic cells may be a protective mechanism in early stages of the disease.
Asunto(s)
Péptidos beta-Amiloides/metabolismo , Monocitos/metabolismo , Fragmentos de Péptidos/metabolismo , Línea Celular , Humanos , Microglía/metabolismo , Placa Amiloide/metabolismoRESUMEN
Most of the cases of early-onset familial Alzheimer's disease (FAD) are related to missense mutations in the presenilin 1 (PS-1) gene on chromosome 14. Although PS-1 mutations are distributed throughout the entire open reading frame, most mutations are found in transmembrane region II and hydrophilic loop VI encoded by exons 5 and 8, respectively. These two groups of substitutions are associated with an age of onset of 40-43 years for exon 5 and 45-55 years for exon 8, respectively. We have previously described a South American pedigree from Argentina with early-onset FAD (mean age of onset 38.9 +/- 3.9 years) with no mutations in exons 16 and 17 of the beta-protein precursor gene (betaPP770 transcript). Here we report the identification of an A --> T transversion at the first position of codon 146 of PS-1 in these patients. This missense mutation results in a Met --> Leu substitution, as reported for the Italian pedigrees Tor1.1 and FAD4. The significant differences in ages of onset and death among members of generations II-III and IV suggest that other genetic and/or environmental factors may influence disease phenotype in this pedigree.
Asunto(s)
Adenina , Enfermedad de Alzheimer/genética , Variación Genética , Leucina/genética , Proteínas de la Membrana/genética , Metionina/genética , Timina , Adulto , Edad de Inicio , Sustitución de Aminoácidos , Argentina , Femenino , Humanos , Masculino , Persona de Mediana Edad , Linaje , Presenilina-1RESUMEN
Estudos de diagnostico, caracterizacao parasitaria e identificacao foram conduzidos em pacientes humanos com lesoes cutaneas de leishmaniose na Provincia de Santiago del Estero, no norte da Argentina. Os procedimentos de diagnostico foram: biopsias de lesoes para utilizacao em esfregacos e inoculacao em hamster; aspiracao (com agulha) de ulceras, para cultura "in vitro". As tecnicas a IFAT-IgG e o teste intradermico de Montenegro. Oito cepas de parasitas foram isoladas, sendo estas obtidas de pacientes com lesoes ativas...
Asunto(s)
Humanos , Anticuerpos Monoclonales/inmunología , Leishmania braziliensis/aislamiento & purificación , Leishmaniasis Cutánea/epidemiología , Anticuerpos Monoclonales/análisis , Argentina , Isoenzimas/análisis , Leishmaniasis Cutánea/diagnóstico , Leishmaniasis Cutánea/inmunologíaRESUMEN
Diagnostic and parasite characterization and identification studies were carried out in human patients with cutaneous leishmaniasis lesions in Santiago del Estero, Northern Province of Argentina. Diagnostic procedures were biopsies of lesions for smears and inoculations in hamster, needle aspirations of material from ulcers for "in vitro" cultures. Immunodiagnostic techniques applied were IFAT-IgG and Montenegro skin test. Primary isolation of eight stocks of leishmanial parasites was achieved from patients with active lesions. All stocks were biologically characterized by their behaviour in hamster, measurements of amastigote and promastigotes and growth "in vitro". Eight stocks were characterized and identified at species level by their reactivity to a cross-panel of sub-genus and species-specific Monoclonal Antibodies through an Indirect Immunofluorescence technique and a Dot-ELISA. We conclude from the serodeme analysis of Argentina stocks that: stocks MHOM/AR/92/SE-1; SE-2; SE-4; SE-8; SE-8-I; SE-30; SE-34 and SE-36 are Leishmania (Viannia) braziliensis. Three Leishmania stocks (SE-1; SE-2 and SE-30) did not react with one highly species-specific Monoclonal Antibody (Clone: B-18, Leishmania-(Viannia) braziliensis marker) disclosing two serodeme group patterns. Five out of eight soluble extracts of leishmanial promastigotes were electrophoresed on thin-layer starch gels and examined for the enzyme MPI, Mannose Phosphate Isomerase; MDH, Malate Dehydrogenase; 6PGD, 6 Phosphogluconate Dehydrogenase; NH, Nucleoside Hydrolase, 2-deoxyinosine as substrate; SOD, Superoxide Dismutase; GPI, Glucose Phosphate Isomerase and ES, Esterase. From the isoenzyme studies we concluded that stocks: MHOM/AR/92/SE-1; SE-2; SE-4; SE-8 and SE-8-I are isoenzymatically Leishmania (Viannia) braziliensis. We need to analyze more enzymes before assigning them to a braziliensis zymodeme.