RESUMEN
BACKGROUND: The optimal choice of conduit and configuration for coronary artery bypass grafting (CABG) in diabetic patients remains somewhat controversial, even though arterial grafts have been proposed as superior. We attempted to clarify the role of complete arterial revascularization using the left internal thoracic artery (LITA) and the radial artery (RA) alone in "T-Graft" configuration on long-term outcome. METHODS AND RESULTS: From 1994 to 2001, 104 diabetic patients with triple vessel disease underwent CABG using LITA/RA "T-Grafts" (Group-A). Using propensity-score matching, 104 patients with comparable preoperative characteristics who underwent CABG using LITA and one sequential vein graft were identified (Group-V). Freedom from all causes of death, cardiac death, major adverse cardiac event (MACE), major adverse cardiac (and cerebral) event (MACCE), and repeat revascularization at 10 years of Group-A was 60 ± 5%, 67 ± 5%, 48 ± 5%, 37 ± 5%, and 81 ± 4%, respectively, compared with 58 ± 5%, 70 ± 5%, 49 ± 5%, 39 ± 5%, and 93 ± 3% in Group-V. There were no significant differences in these end points between groups regardless of insulin-dependency. Multivariable Cox proportional hazards model identified age, left ventricular ejection fraction, renal failure, and hyperlipidemia as independent predictors for all death, age and left ventricular ejection fraction for cardiac death, sinus rhythm for both MACE and MACCE, and prior percutaneous coronary intervention for re-revascularization. CONCLUSIONS: In our experience, complete arterial revascularization using LITA/RA "T-Grafts" does not provide superior long-term clinical benefits for diabetic patients compared with a combination of LITA and sequential vein graft.
Asunto(s)
Puente de Arteria Coronaria/métodos , Enfermedad de la Arteria Coronaria/cirugía , Diabetes Mellitus , Arterias Mamarias/cirugía , Arteria Radial/trasplante , Vena Safena/trasplante , Anciano , Puente de Arteria Coronaria/efectos adversos , Puente de Arteria Coronaria/mortalidad , Enfermedad de la Arteria Coronaria/mortalidad , Diabetes Mellitus/mortalidad , Femenino , Humanos , Masculino , Persona de Mediana Edad , Complicaciones Posoperatorias/mortalidad , Complicaciones Posoperatorias/cirugía , Supervivencia sin Progresión , Reoperación , Estudios Retrospectivos , Medición de Riesgo , Factores de Riesgo , Factores de TiempoRESUMEN
Aseptic cell sorting is challenging, especially when a flow-cytometric cell sorter is not operated in a sterile environment. The sheath fluid system of a cell sorter may be contaminated with germs such as bacteria, yeasts, viruses, or fungi. Thus, a regular chemical cleaning procedure is required to prepare a sorter for aseptic cell sorting by flushing the fluidic system. However, this procedure is time consuming, and most importantly, the researcher can never be sure that the cleaning process was successful. Here we present a method in which the sheath fluid of a cell sorter was decontaminated by irradiation with UV-C light using a flow-through principle. Using this principle, we were able to achieve a 5 log reduction of bacteria in the sheath fluid.
RESUMEN
Flow cytometers are robust and ubiquitous tools of biomedical research, as they enable high-throughput fluorescence-based multi-parametric analysis and sorting of single cells. However, analysis is often constrained by the availability of detection reagents or functional changes of cells caused by fluorescent staining. Here, we introduce MAPS-FC (multi-angle pulse shape flow cytometry), an approach that measures angle- and time-resolved scattered light for high-throughput cell characterization to circumvent the constraints of conventional flow cytometry. In order to derive cell-specific properties from the acquired pulse shapes, we developed a data analysis procedure based on wavelet transform and k-means clustering. We analyzed cell cycle stages of Jurkat and HEK293 cells by MAPS-FC and were able to assign cells to the G1, S, and G2/M phases without the need for fluorescent labeling. The results were validated by DNA staining and by sorting and re-analysis of isolated G1, S, and G2/M populations. Our results demonstrate that MAPS-FC can be used to determine cell properties that are otherwise only accessible by invasive labeling. This approach is technically compatible with conventional flow cytometers and paves the way for label-free cell sorting.
Asunto(s)
Ciclo Celular , Citometría de Flujo/instrumentación , Células HEK293 , Humanos , Células JurkatRESUMEN
The tumor vaccine MGN1601 was designed and developed for treatment of metastatic renal cell carcinoma (mRCC). MGN1601 consists of a combination of fourfold gene-modified cells with the toll-like receptor 9 agonist dSLIM, a powerful connector of innate and adaptive immunity. Vaccine cells originate from a renal cell carcinoma cell line (grown from renal cell carcinoma tissue), express a variety of known tumor-associated antigens (TAA), and are gene modified to transiently express two co-stimulatory molecules, CD80 and CD154, and two cytokines, GM-CSF and IL-7, aimed to support immune response. Proof of concept of the designed vaccine was shown in mice: The murine homologue of the vaccine efficiently (100%) prevented tumor growth when used as prophylactic vaccine in a syngeneic setting. Use of the vaccine in a therapeutic setting showed complete response in 92% of mice as well as synergistic action and necessity of the components. In addition, specific cellular and humoral immune responses in mice were found when used in an allogeneic setting. Immune response to the vaccine was also shown in mRCC patients treated with MGN1601: Peptide array analysis revealed humoral CD4-based immune response to TAA expressed on vaccine cells, including survivin, cyclin D1, and stromelysin.
RESUMEN
Gene modification of eukaryotic cells by electroporation is a widely used method to express selected genes in a defined cell population for various purposes, like gene correction or production of therapeutics. Here, we describe the generation of a cell-based tumor vaccine via fourfold transient gene modification of a human renal cell carcinoma (RCC) cell line for high expression of CD80, CD154, GM-CSF, and IL-7 by use of MIDGE(®) vectors. The two co-stimulatory molecules CD80 and CD154 are expressed at the cell surface, whereas the two cytokines GM-CSF and IL-7 are secreted yielding cells with enhanced immunological properties. These fourfold gene-modified cells have been used as a cell-based tumor vaccine for the treatment of RCC.