GATA2 zinc finger 2 mutation found in acute myeloid leukemia impairs myeloid differentiation

AbstractWe identified two novel GATA2 mutations in acute myeloid leukemia (AML). One mutation (p.R308P-GATA2) was a R308P substitution within the zinc finger (ZF)-1 domain, and the other (p.A350_N351ins8-GATA2) was an eight-amino-acid insertion between A350 and N351 residues within the ZF-2 domain. p.R308P-GATA2 did not affect DNA-binding and transcriptional activities, while p.A350_N351ins8-GATA2 reduced them, and impaired G-CSF-induced granulocytic differentiation of 32D cells. Although p.A350_N351ins8-GATA2 did not show a dominant-negative effect over wild-type (Wt)–GATA2 by the reporter assay, it might be involved in the pathophysiology of AML by impairing myeloid differentiation because of little Wt-GATA2 expression in primary AML cells harboring the p.A350_N351ins8 mutation

No relationship exists between PBP 2a amounts expressed in different MRSA strains obtained clinically and their β-lactam MIC values

After establishing a linear relationship between the amount of penicillinbinding protein (PBP) 2a and membrane proteins of methicillin-resistant Staphylococcus aureus (MRSA) COL by dot-blot analysis using an antibody against PBP 2a, we determined the PBP 2a quantities in membrane fractions prepared from 14 different MRSA cells. Methicillin-sensitive S. aureus ATCC 6538P was used as a quality control strain. The amounts of PBP 2a diverged among the strains, and no relationship to β-lactam MIC values were observed in the corresponding strains

In Vitro Endothelialization Test of Biomaterials Using Immortalized Endothelial Cells.

Functionalizing biomaterials with peptides or polymers that enhance recruitment of endothelial cells (ECs) can reduce blood coagulation and thrombosis. To assess endothelialization of materials in vitro, primary ECs are generally used, although the characteristics of these cells vary among the donors and change with time in culture. Recently, primary cell lines immortalized by transduction of simian vacuolating virus 40 large T antigen or human telomerase reverse transcriptase have been developed. To determine whether immortalized ECs can substitute for primary ECs in material testing, we investigated endothelialization on biocompatible polymers using three lots of primary human umbilical vein endothelial cells (HUVEC) and immortalized microvascular ECs, TIME-GFP. Attachment to and growth on polymer surfaces were comparable between cell types, but results were more consistent with TIME-GFP. Our findings indicate that TIME-GFP is more suitable for in vitro endothelialization testing of biomaterials

Endothelialization effects of biocompatible polymers, as measured using TIME-GFP.

<p>Endothelialization effects of biocompatible polymers, as measured using TIME-GFP.</p

HUVEC attachment and growth profiles on various polymer surfaces.

<p>Cells (6 × 10⁴) were seeded on polymer-coated PC discs and the number of cells was counted 1 and 4 days after seeding. Results of three lots of HUVEC (A, B, and C) are presented. Error bars are standard deviations (SD) of the triplicate samples.</p

Quantitative analysis of vascular EC marker gene expression.

<p>(A) <i>NOS3</i> and <i>TM</i> expression in TIME-GFP and three lots of HUVEC grown in a culture dish for 4 days, as determined by dPCR. Results are mean ± SD of three independent experiments with TIME-GFP and three lots of HUVECs, with expression in TIME-GFP set as 1. (B) <i>NOS3</i> and <i>TM</i> expression in TIME-GFP and three lots of HUVEC grown on various polymer surfaces for 4 days, as determined by dPCR. Data are mean ± SD of three independent experiments, with expression in dish cultures set as 1. Statistical significance was determined by one-way ANOVA and Bonfferroni’s post-hoc test. *<i>P</i> < 0.05 vs dish culture; n.d., not determined due to insufficient total RNA.</p

Attachment profiles of TIME-GFP on discs with 100%, 50%, and 0% PMEA coverage.

<p><i>Y</i>-axis indicates the number of cells after 4 days of culture. Data are mean ± SD of three independent experiments. Statistical significance was determined by one-way non-repeated measures analysis of variance and the Student-Newman-Keuls’s post-hoc test (*<i>P</i> < 0.05).</p

Phase contrast microscopy of HUVECs on various polymer surfaces.

<p>(A, B) Cells (6 × 10⁴) were seeded on polycarbonate (PC) discs coated with the indicated polymers (= 33 μm, thickness = 0.1 mm). The discs were placed in PMPC-coated 6-well plates. Representative images of HUVEC-A at 1 day (A) and 4 days (B) after seeding are shown. Scale bars = 300 μm. Untreat, untreated PC disc; PMPC, no PC disc.</p

Screening study on hemolysis suppression effect of an alternative plasticizer for the development of a novel blood container made of polyvinyl chloride

Abstract: The aim of this study is to identify a plasticizer that is effective in the suppression of the autohemolysis of the stored blood and can be used to replace di(2-ethylhexyl) phthalate (DEHP) in blood containers. The results of hemolysis test using mannitol-adenine-phosphate/red cell concentrates (MAP/RCC) spiked with plasticizers included phthalate, phthalate-like, trimeliate, citrate, and adipate derivatives revealed that di-isononyl-cyclohexane-1,2-dicarboxylate (Hexamoll , and diisodecyl phthalate (DIDP) exhibited a hemolysis suppression effect almost equal to that of DEHP, but not other plasticizers. This finding suggested that the presence of 2 carboxy-ester groups at the ortho position on a 6-membered ring of carbon atoms may be required to exhibit such an effect. The hemolytic ratios of MAP/RCC-soaked polyvinyl chloride (PVC) sheets containing DEHP or different amounts of DINCH or DOTP were reduced to 10.9%, 9.2-12.4%, and 5.2-7.8%, respectively (MAP/RCC alone, 28.2%) after 10 weeks of incubation. The amount of plasticizer eluted from the PVC sheet was 53.1, 26.1-36.5, and 78.4-150 mg/mL for DEHP, DINCH, and DOTP, respectively. PVC sheets spiked with DIDP did not suppress the hemolysis induced by MAP/ RCC because of low leachability (4.8-6.0 mg/mL). These results suggested that a specific structure of the plasticizer and the concentrations of least more than 10 mg/mL were required to suppress hemolysis due to MAP/RCC