RESUMEN
OBJECTIVE: In view of the limitations of albumin in peritoneal dialysis (PD), we set out to evaluate whether total lymphocyte counts (TLC) could serve as a better prognostic indicator. We were also interested in how these parameters might differ between PD and hemodialysis (HD) patients. DESIGN: In a retrospective study, we reviewed 113 charts from our dialysis unit. All laboratory analyses were performed by the Department of Clinical Pathology of the Nassau County Medical Center, using standard procedures. Intact parathyroid hormone (PTH) was sent out to Nichols Laboratories. SETTING: All patients originated from the renal clinic at Nassau County Medical Center, a 612 bed public hospital. PATIENTS: The 38 PD and 75 HD patients selected had been receiving dialysis for at least 12 months and up to 3 years. The PD patients received either continuous ambulatory and/or cycler PD. For the survivors, the averages of their routine chemical analyses were considered their representative values. For the nonsurvivors, the most recent laboratory values prior to their end point were considered. MAIN OUTCOME MEASURES: Mortality or apparent malnutrition leading to transfer to HD represented the end points for PD patients. Mortality alone was used as the end point for HD patients. RESULTS: Within the PD population, serum albumin was not significantly lower in nonsurvivors compared to survivors, while the TLC was significantly lower in nonsurvivors (1277 +/- 146/mm3 vs 2249 +/- 236/mm3, p = 0.0036). The HD population demonstrated a significant difference in both TLC and serum albumin levels between its two prognostic groups; albumin was the better discriminator. Nonsurvivors had a 20% lower serum albumin than did the survivors (27.0 +/- 1.6 g/L vs 34.0 +/- 0.5 g/L, p = 0.0001). Patients on PD had a higher TLC than those on HD (p = 0.0001). CONCLUSIONS: In the HD population, but not in the PD population, both serum albumin and TLC were significantly higher in the group that survived. Serum albumin is a more powerful discriminator of mortality in the HD population, while TLC is a better discriminator of mortality in the PD population. For uncertain reasons, PD patients have a higher TLC than those on HD.
Asunto(s)
Fallo Renal Crónico/sangre , Fallo Renal Crónico/mortalidad , Diálisis Peritoneal Ambulatoria Continua , Diálisis Renal , Femenino , Humanos , Fallo Renal Crónico/terapia , Recuento de Linfocitos , Masculino , Persona de Mediana Edad , Pronóstico , Estudios Retrospectivos , Tasa de SupervivenciaRESUMEN
Gene transfer by direct intramuscular injection of naked plasmid DNA has been shown to be a safe, simple but relatively inefficient method for gene delivery in vivo. Eukaryotic plasmid expression vectors incorporating the Epstein-Barr virus (EBV) origin of replication (oriP) and EBNA1 gene have been shown to act as autonomous episomally replicating gene transfer vectors which additionally provide nuclear matrix retention functions. Prolonged expression of a LacZ reporter gene and recombinant human dystrophin was shown using EBV-based plasmid vectors transfected into C2C12 mouse myoblast and myotube cultures. Intramuscular injection of EBV-based dystrophin expression plasmids into nude/mdx mice resulted in significant enhancement in the number of muscle fibres expressing recombinant dystrophin compared with a conventional vector. This effect was observed for over 10 weeks after a single administration. These results indicate the potential advantage of EBV-based expression vectors for focal plasmid-mediated gene augmentation therapy in Duchenne muscular dystrophy (DMD) and a range of other gene therapeutic applications.
Asunto(s)
Cromosomas , Distrofina/genética , Terapia Genética/métodos , Vectores Genéticos/administración & dosificación , Herpesvirus Humano 4/genética , Distrofia Muscular de Duchenne/terapia , Animales , Expresión Génica , Vectores Genéticos/genética , Humanos , Inyecciones Intramusculares , Ratones , Ratones Endogámicos mdx , Plásmidos , Factores de TiempoRESUMEN
Duchenne muscular dystrophy (DMD) is an X-linked, lethal disease caused by mutations of the dystrophin gene. No effective therapy is available, but dystrophin gene transfer to skeletal muscle has been proposed as a treatment for DMD. We have developed a strategy for efficient in vivo gene transfer of dystrophin cDNA into regenerating skeletal muscle. Retroviral producer cells, which release a vector carrying the therapeutically active dystrophin minigene, were mitotically inactivated and transplanted in adult nude/mdx mice. Transplantation of 3 x 10(6) producer cells in a single site of the tibialis anterior muscle resulted in the transduction of between 5.5 and 18% total muscle fibers. The same procedure proved also feasible in immunocompetent mdx mice under short-term pharmacological immunosuppression. Minidystrophin expression was stable for up to 6 mo and led to alpha-sarcoglycan reexpression. Muscle stem cells could be transduced in vivo using this procedure. Transduced dystrophic skeletal muscle showed evidence of active remodeling reminiscent of the genetic normalization process which takes place in female DMD carriers. Overall, these results demonstrate that retroviral-mediated dystrophin gene transfer via transplantation of producer cells is a valid approach towards the long-term goal of gene therapy of DMD.
Asunto(s)
Distrofina/deficiencia , Distrofina/genética , Terapia Genética , Músculo Esquelético/fisiopatología , Distrofia Muscular Animal/terapia , Animales , Trasplante de Células , ADN Complementario/genética , Femenino , Técnicas de Transferencia de Gen , Vectores Genéticos , Inmunohistoquímica , Ratones , Ratones Endogámicos mdx , Distrofia Muscular Animal/genética , Distrofia Muscular Animal/fisiopatología , RetroviridaeAsunto(s)
Distrofina/biosíntesis , Terapia Genética/métodos , Distrofias Musculares/terapia , Distrofia Muscular Animal/terapia , Transfección/métodos , Animales , Distrofina/genética , Genes Reporteros , Vectores Genéticos , Humanos , Kanamicina Quinasa , Operón Lac , Ratones , Ratones Endogámicos C57BL , Ratones Endogámicos mdx , Ratones Desnudos , Distrofias Musculares/genética , Fosfotransferasas (Aceptor de Grupo Alcohol)/biosíntesis , RetroviridaeRESUMEN
The humoral immune response of 34 macaques experimentally infected with SIVmac-251 was studied using a combination of an epitope library and synthetic peptides. The course of the immune response was checked for up to 9 months postinfection with a panel of clones expressing SIV fragments. A systematic study was performed with synthetic peptides covering the whole transmembrane (TM) and external (SU) envelope proteins. Seven major immunodominant epitopes were characterized. Four are localized in the SU protein: one in the V1 region (111-130), one in the Cys loop of the V3 region (311-330) and two in the C-terminal end (501-520 and 511-530). Three are localized in the TM protein: one in the extracellular domain (601-619), one in the anchor domain (731-750) and one in the intracytoplasmic domain (861-881). Among these epitopes, only one, 601-619, was found to be reactive with all sera and can be defined as the principal immunodominant epitope.
Asunto(s)
Linfocitos B/inmunología , Epítopos Inmunodominantes , Síndrome de Inmunodeficiencia Adquirida del Simio/inmunología , Proteínas del Envoltorio Viral/inmunología , Secuencia de Aminoácidos , Animales , Linfocitos B/virología , Secuencia de Bases , Clonación Molecular , Cartilla de ADN , Modelos Animales de Enfermedad , Ensayo de Inmunoadsorción Enzimática , Macaca mulatta , Datos de Secuencia Molecular , Síndrome de Inmunodeficiencia Adquirida del Simio/virologíaRESUMEN
Genomic DNA of Toxoplasma gondii was digested with the restriction endonuclease Hpa II and the resulting repetitive DNA sequences were visualized after electrophoresis on agarose gels and staining with ethidium bromide. Three repetitive DNA sequences were isolated and cloned in the plasmid pUC19. The recombinant plasmids (pTg8, pTg4 and pTg1) had inserts of 840, 440, and 180 basepairs, respectively. The estimated copy number of these cloned sequences in the T. gondii genome was approximately 800-1,000 for pTg4, 150-200 for pTg8, and 30-40 for pTg1. In dot-blot hybridization tests, pTg4 was able to detect as little as 80 pg of purified T. gondii DNA or 1,000 parasites in the presence or absence of 1.5 x 10(6) human or mouse leukocytes. No cross-hybridization was detected with 10 micrograms of either human and mouse DNA or 100 ng of DNA from other parasites (Eimeria tenella, E. acervularia, Trypanosoma cruzi, and Leishmania donovani), or among the three DNA probes. Each probe identified T. gondii tachyzoites in tissue (liver and spleen) obtained from experimentally infected mice in which histologic damage was detected. In addition, early detection of T. gondii in brain tissue and blood samples was possible with the pTg4 probe.