RESUMEN
The aim of this study was to evaluate the total phenolic content, composition, and antioxidant and antibacterial activities of four grape seed extracts (Cabernet Sauvignon, Marselan, Pinot Noir, and Tamyanka). The total phenolic content (TPC) and flavonoid, anthocyanin, procyanidin, ascorbic acid, DPPH, and ABTS antioxidant capacities of the grape seed extracts (GSEs) were determined. The extracts showed high TPC values (79.06-111.22 mg GAE/g). The individual components in the GSEs were determined using HPLC. High contents of catechin, epicatechin, and procyanidin B1 were found in the extracts. The antimicrobial activity of the obtained GSEs against Staphylococcus aureus, Bacillus cereus, and Escherichia coli was evaluated using the agar diffusion test and a test to determine the minimum inhibitory concentration (MIC). According to the effect on the growth of pathogens, the extracts were ranked in the following order: Pinot Noir > Marselan > Cabernet Sauvignon > Tamyanka. The tested bacteria showed high sensitivity to the extracts (MIC = 0.12-0.50 mg/mL). According to the MIC values, the bacteria were in the following order: S. aureus > B. cereus > E. coli. A correlation was found between the phenolic content of the GSEs and their antibacterial potential. The obtained results show that the studied GSEs have good potential as antioxidant and antimicrobial agents.
RESUMEN
Accurate counting of CD34-positive cells is important for successful hematopoietic stem cell transplantation that is applied to various diseases. The aim of this study was simultaneous counting of viable CD34+ (vCD34+) and CD45+ (vCD45+) cells in apheresis samples by automatic immunofluorescence counter - EasyCounter BC. CD34+ and CD45+ cells were counted using two conjugates anti-CD34 antibody - dR110 and anti-CD45 antibody - ATTO620, respectively. The conjugates were prepared by carbodiimide method. Dead nuclear cells were counted by using monomethine cyanine dye PO-TEDM 1. The linearity and reproducibility of EasyCounter BC for CD34+ cell counting were determined (R2 = 0.99; CV values for vCD34+ cells were 6.8 ÷ 8.5% and for vCD45+ cells 4.1 ÷ 7.2%). The obtained results by EasyCounter BC were compared with those by other two standard methods - flow cytometry (Guava easyCyte 8HT) and fluorescence microscopic method (Olympus BX51) with the same conjugates. Passing-Bablok regression was performed to determine the relationship between the results of the three methods, analyzing 43 apheresis samples. Correlation coefficients for vCD45+ and vCD34+ between EasyCounter BC and Olympus microscope were 0.987 and 0.982, respectively (P < 0.0001). Better results were obtained between EasyCounter BC and flow cytometer Guava, 0.998 for vCD45+ and 0.998 for vCD34+ (P < 0.0001).
Asunto(s)
Antígenos CD34/química , Citometría de Imagen , Antígenos Comunes de Leucocito/química , Leucocitos/citología , Células Madre/citología , Autoanálisis , Citometría de Flujo , Humanos , Microscopía Fluorescente , Coloración y EtiquetadoRESUMEN
The ability of immobilized conjugate anti-CD34+ monoclonal antibody-dR110 and free conjugate anti-CD45+ monoclonal antibody-ATTO620 to precisely enumerate CD34+ stem cells and CD45+ cells in apheresis samples were evaluated. The conjugates anti-CD34+ antibody-dR110 and anti-CD45+- antibody-ATTO620 were prepared. Functionalized magnetic nanoparticles (MNPs) were synthesized. The anti-CD34+ antibody-dR110 conjugate was immobilized on the modified MNPs using a carbodiimide method. The stem cell count in thawed apheresis samples was determined using the free and the immobilized conjugate anti-CD34+ antibody-dR110 on MNPs and an image cell counter EasyCounter BC. A higher stem cell count and more accurate results were obtained with the immobilized conjugate, because a separation and concentration of the stem cells bound to antibody-dR110 on MNPs by external magnet were performed. Coefficients of variation of CD34+ cell count in apheresis samples, determined by EasyCounter BC, were ranged from 5.5 to 6.9% and those of CD45+ cell count from 3.8 to 4.7%. The viability of CD34+ cells was high from 98.5 to 99.6%. It was found that correlation coefficient between the flow cytometer and automatic cell counter, using free anti-CD34+ antibody-dR110 was 0.94, and when using immobilized anti-CD34+antibody-dR110 on MNPs, the correlation coefficient was 0.97.