The Effect of Heterozygous Mutation of Adenylate Kinase 2 Gene on Neutrophil Differentiation

Mitochondrial ATP production plays an important role in most cellular activities, including growth and differentiation. Previously we reported that Adenylate kinase 2 (AK2) is the main ADP supplier in the mitochondrial intermembrane space in hematopoietic cells, especially in the bone marrow. AK2 is crucial for the production of neutrophils and T cells, and its deficiency causes reticular dysgenesis. However, the relationship between ADP supply by AK2 and neutrophil differentiation remains unclear. In this study, we used CRISPR/Cas9 technology to establish two heterozygous AK2 knock-out HL-60 clones as models for reticular dysgenesis. Their AK2 activities were about half that in the wild-type (WT). Furthermore, neutrophil differentiation was impaired in one of the clones. In silico analysis predicted that the obtained mutations might cause a structural change in AK2. Time course microarray analysis of the WT and mutants revealed that similar gene clusters responded to all-trans retinoic acid treatment, but their expression was lower in the mutants than in WT. Application of fructose partially restored neutrophil differentiation in the heterozygous knock-out HL-60 clone after all-trans retinoic acid treatment. Collectively, our study suggests that the mutation of N-terminal region in AK2 might play a role in AK2-dependent neutrophil differentiation and fructose could be used to treat AK2 deficiency

Indomethacin誘発性腸管傷害ラットにおける血漿α-1酸性糖タンパク質（AGP）の変動

ラットに Indomethacin（Indo）を投与するとクローン病（CD）様の腸管傷害がみられる。本モデルにおいて血漿タンパク質の SDS-PAGE 解析をおこなったところ、増加するタンパク質が認められた。そこで本研究ではこれらのタンパク質を同定し、炎症との関連を検討することを目的とした。Indo 投与後の経日的変化を観察した結果、腸管傷害は投与2、3日目で重篤となり、同時に 37～50kDa のタンパク質に増加がみられた。これらのタンパク質は PAS 染色陽性であり、糖タンパク質であった。これらの特徴より推測し、ウエスタンブロッティングを行った結果、α-1 酸性糖タンパク質（AGP）であることを明らかにした。また、AGP は血中のみならず、肝臓や小腸においても増加しており、炎症による産生亢進が示唆された。AGP は CRP に比べて血中濃度が高く、CBB 染色で容易に判別できるため、実験動物等における炎症状態の判定にも利用できると考えられる

Association between Preterm Birth and Vaginal Colonization by Mycoplasmas in Early Pregnancy

To examine the association between colonization by two newly classified species of genital ureaplasmas (Ureaplasma parvum and U. urealyticum) in early pregnancy and subsequent late abortion or preterm birth at <34 weeks of gestation, four species of genital mycoplasmas—Mycoplasma genitalium, M. hominis, U. parvum, and U. urealyticum—as well as Chlamydia trachomatis and Neisseria gonorrhoeae were examined by PCR-based methods in a prospective cohort study of 877 women with singleton pregnancies at <11 weeks of gestation. Antibiotics were used only in cases in which C. trachomatis and/or N. gonorrhoeae was detected. Multivariate logistic-regression analysis was used to assess independent risk factors after taking maternal low body weight and past history of preterm birth into account. M. genitalium, M. hominis, U. parvum, U. urealyticum, C. trachomatis, and N. gonorrhoeae were detected in 0.8%, 11.2%, 52.0%, 8.7%, 3.2%, and 0.1% of these 877 women, respectively. Twenty-one (2.4%) women experienced late abortion or preterm birth at <34 weeks of gestation. Three factors—detection of U. parvum in the vagina (odds ratio [OR], 3.0; 95% confidence interval [CI], 1.1 to 8.5); use of antibiotics, such as penicillin and cefatrizine, for incidental inflammatory complications before 22 weeks of gestation (OR, 4.2; 95% CI, 1.6 to 10.0); and past history of preterm birth (OR, 10.4; 95% CI, 2.7 to 40.5)—were independently associated with late abortion and preterm birth. In conclusion, vaginal colonization with U. parvum, but not U. urealyticum, is associated with late abortion or early preterm birth

Down‐regulation of α6 integrin, an anti‐oncogene product, by functional cooperation of H‐Ras and c‐Myc

Background The molecular basis of cooperation of H‐Ras and c‐Myc in regulating cellular behaviour, such as cell adhesiveness, is still poorly understood. To investigate the role of H‐Ras and c‐Myc in cell adhesiveness, a constitutively active H‐RasV12 (H‐RasV12) and c‐Myc were stably expressed, singly or in combination in a haematopoietic cell line, and the expression and activity of cell adhesion molecules were monitored. Results We have shown that the ectopic expression of H‐RasV12, but not c‐Myc alone, in a haematopoietic cell line, induces the activation of very late antigen‐6 (VLA‐6, α6β1) integrin. Co‐expression of H‐RasV12 and c‐Myc in the same cells further resulted in the induction of expression of vascular cell adhesion molecule‐1 (VCAM‐1) and the inhibition of expression of α6 integrin, a candidate anti‐oncogene product, leading to a loss of adhesiveness to laminin (Lm), a ligand for VLA‐6. Conclusions Cooperation of H‐Ras and c‐Myc reciprocally regulates expression of the adhesion molecules, α6 integrin and VCAM‐1. Our results represent an unprecedented account of the cooperation of the oncogene products, H‐Ras and c‐Myc, to inhibit expression of an anti‐oncogene product, α6 integrin