RESUMO
Accumulating lines of clinical evidence support the emerging hypothesis that loss-of-function mutations of GATA2 cause inherited hematopoietic diseases, including Emberger syndrome; dendritic cell, monocyte B and NK lymphoid (DCML) deficiency; and MonoMAC syndrome. Here, we show that mice heterozygous for an arginine-to-tryptophan substitution mutation in GATA2 (G2R398W/+), which was found in a patient with DCML deficiency, substantially phenocopy human DCML deficiency. Mice heterozygous for the GATA2-null mutation (G2-/+) do not show such phenotypes. The G2R398W protein possesses a decreased DNA-binding affinity but obstructs the function of coexpressed wild-type GATA2 through specific cis-regulatory regions, which contain two GATA motifs in direct-repeat arrangements. In contrast, G2R398W is innocuous in mice containing single GATA motifs. We conclude that the dominant-negative effect of mutant GATA2 on wild-type GATA2 through specific enhancer/silencer of GATA2 target genes perturbs the GATA2 transcriptional network, leading to the development of the DCML-like phenotype. The present mouse model provides an avenue for the understanding of molecular mechanisms underlying the pathogenesis of GATA2-related hematopoietic diseases.
Assuntos
Monócitos , Receptores de Antígenos Quiméricos , Animais , Fator de Transcrição GATA2/genética , Heterozigoto , Humanos , Camundongos , Monócitos/metabolismo , Mutação , Fenótipo , Ligação Proteica , Receptores de Antígenos Quiméricos/genéticaRESUMO
OBJECTIVES: To clarify the characteristics and growth of bacteria that may infiltrate liquid baby formula during feeding and after storage for more than 3 h, the transfer of oral bacteria through artificial nipples, and bacterial survival in liquid baby formula and a baby drink were examined immediately after drinking and after storage at 4 °C for 12 h and 24 h. METHODS: Thirteen human subjects (aged 19-24 years) were asked to drink approximately 50 mL of liquid baby formula and a baby drink, via the artificial nipple of a baby bottle. Samples of the remaining liquid after storage at 4 °C for 12 h and 24 h were inoculated onto blood agar plates and incubated anaerobically at 37 °C for 7 days. Genomic DNA was extracted from individual colonies, and the bacterial species were identified by 16S rRNA gene sequencing. RESULTS: The mean concentrations of bacteria in the liquid baby formula were (2.6 ± 2.8) × 104 and (4.1 ± 6.6) × 104 colony-forming unit/mL after storage at 4 °C for 12 h and 24 h, respectively. Streptococcus (43.2%), Veillonella (9.3%), and Schaalia (8.2%) species were recovered from the remaining liquid baby formula after storage at 4 °C for 12 h. In contrast, no bacteria were detected in the remaining baby drink after storage at 37 °C for 24 h. CONCLUSIONS: The levels of bacteria immediately after drinking and after storage at 4 °C for 12 h or 24 h were similar, suggesting that remaining liquid baby formula may be preserved safely in a refrigerator for more than 3 h.
Assuntos
Microbiota , Mamilos , Bactérias/genética , Humanos , Fórmulas Infantis , RNA Ribossômico 16S/genéticaRESUMO
Aspergillus oryzae KB produces two ß-fructofuranosidases (F1 and F2). F1 has high transferring activity and produces fructooligosaccharides from sucrose. Mycelial growth pellets were altered by the addition of Tween 20, 40 and 80 (HLB=16.7, 15.6 and 15.0, respectively) in liquid medium cultures to form small spherical pellets. The particle size of the pellets decreased with the HLB value, which corresponds to an increase in surfactant hydrophobicity. Selective F1 production and pellet size were maximized using Tween 20. Adding polyoxyethylene oleyl ethers (POEs) with various degrees of polymerization (2, 7, 10, 20 and 50: HLB=7.7, 10.7, 14.7, 17.2 and 18.2, respectively) was investigated. A minimum mean particle size was obtained using a POE with DP=10, HLB=14.7. The POE surfactants had little effect on the selective production of F1. The formation of filamentous pellets depended on the surfactant HLB value, and F1 enzymes were produced most efficiently using Tween 20.