In this study, we evaluated the expression of molecular markers of acute rejection in protocol biopsies of patients with and without subclinical acute rejection (SAR). Protocol biopsies were performed at 2 months (n = 21) and 12 months (n = 14) after kidney transplantation in patients with stable allograft function. After biopsy tissue RNA isolation, reverse transcription and polymerase chain reaction (RT-PCR) for the glyceraldehyde 3-phospate dehydrogenase (GAPDH), perforin, granzyme B and Fas ligand genes were performed. The Banff 97 classification was used for histological diagnosis. Creatinine concentrations at 2 months were significantly higher in patients with SAR (1.46 +/- 0.27 x 1.18 +/- 0.24; p < 0.02). Perforin transcripts were found in 15 biopsy specimens, 10 of which had histological signs of SAR (p = 0.06). Granzyme B expression was found in 10 specimens, nine of which had SAR (p < 0.01). Fas ligand was expressed in seven specimens, and six of them were classified as SAR (p < 0.01). Perforin expression had the highest sensitivity (81%) for the diagnosis of SAR. Granzyme B and Fas ligand had specificity of 90%. At 12 months, there was no significant difference in creatinine concentrations for patients with and without previous SAR (1.63 +/- 0.57 x 1.28 +/- 0.31; p = 0.10). Molecular analysis revealed that there was no statistically significant difference in the expression of perforin and granzyme B in patients with and without SAR. Fas ligand expression was observed in five samples, four of which had histological signs of SAR (p = 0.03). At 12 months, perforin expression had the highest sensitivity (83%), and Fas ligand, the highest specificity (88%) for the diagnosis of SAR. We concluded that the expression of genes that encode proteins involved in the cytolytic attack against the allograft is increased in kidneys with SAR. These findings support the understanding that SAR is an active immune process potentially deleterious to renal allografts.
This article discusses the main principles of infection prevention and control in non-acute healthcare settings. It explores the use of a set of ten tools developed by the Infection Control Nurses Association (ICNA) to audit infection prevention and control, using the standard statements and criteria within the tools as a checklist. The results of the audit of facilities, commodities and practice using the ICNA audit tools will help staff to identify areas of best practice and areas where improvements are needed to enhance patient care.
Objetivo: avaliar a indução de apoptose pela rapamicina em linfócitos humanos periféricos. Métodos: Polimorfos mononucleares foram separados do sangue periférico de voluntários sadios através de centrifugação em gradiente de densidade. A fração mononuclear foi suspensa em meio de cultura, e após, transferida para placas de cultura, às quais acrescentou-se fito-hemaglutinina (PHA) e/ou rapamicina conforme o ensaio. Foram testados quatro ensaios experimentais: cultura de linfócitos puros, linfócitos com PHA, linfócitos com rapamicina, e linfócitos com PHA e rapamicina. As culturas foram incubadas a 37°C em atmosfera estéril, com 5% de CO2 por 24 e 48 horas. A apoptose foi determinada através da marcação com Anexina V por citometria de fluxo. Resultados: não houve diferença estatisticamente significativa na detecção de apoptose em linfócitos com e sem rapamicina, tanto na análise após 24h (7,1% + 3,8 x 6,6% + 2,6, p=1,0) como após 48h ( 6,1% + 1,9 x 6,2 + 1,8, p=1,0 ). Já, linfócitos com PHA, na presença ou ausência da droga, aumentou estatisticamente a apoptose, tanto nas análises de 24h como nas de 48h ( P = 0,0000 e P = 0,0006, ANOVA). Nas culturas estimuladas com PHA a adição de rapamicina também não ocasionou aumento estatisticamente significativo nos percentuais de apoptose tanto em 24h (P=0,69) como após 48h (P>0,73). Conclusão: Os achados deste estudo mostram que a rapamicina não induz apoptose em cultura de linfócitos estimulada ou não.
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