Clone-forming activity of embryonal stem hemopoietic cells after transplantation to newborn or adult sublethally irradiated mice

Hemopoietic activity of stem hemopoietic cells from the liver of embryos was studied at different terms of intrauterine development. The fate of individual clones of hemopoietic cells marked by human adenosine deaminase gene was followed up in sublethally irradiated or newborn recipients. The efficiency of marker gene incorporation in primitive stem hemopoietic cells from the liver of 12-, 13-, and 17-day embryos was not high. Gene transfer was performed without cell prestimulation to division, and hence, these data show that primitive stem cells proliferate even in 17-day embryos. Cells from embryonal liver in all terms maintain hemopoiesis both in newborn and adult microenvironment, hemopoiesis being realized according to the clonal succession model,i. e. in the some way after transplantation of the bone marrow from adult mice. Translated fromByulleten' Eksperimental'noi Biologii i Meditsiny, Vol. 130, No. 7, pp. 110–112, July, 2000     

Clone-forming activity of embryonal stem hemopoietic cells after transplantation to newborn or adult sublethally irradiated mice

Hemopoietic activity of stem hemopoietic cells from the liver of embryos was studied at different terms of intrauterine development. The fate of individual clones of hemopoietic cells marked by human adenosine deaminase gene was followed up in sublethally irradiated or newborn recipients. The efficiency of marker gene incorporation in primitive stem hemopoietic cells from the liver of 12-, 13-, and 17-day embryos was not high. Gene transfer was performed without cell prestimulation to division, and hence, these data show that primitive stem cells proliferate even in 17-day embryos. Cells from embryonal liver in all terms maintain hemopoiesis both in newborn and adult microenvironment, hemopoiesis being realized according to the clonal succession model,i. e. in the some way after transplantation of the bone marrow from adult mice. Translated fromByulleten' Eksperimental'noi Biologii i Meditsiny, Vol. 130, No. 7, pp. 110–112, July, 2000     

The number of functioning hemopoietic clones in transplantation-restored bone marrow of mice depends on the dose of transplanted cells

The clone composition of mouse bone marrow restored by transplantations of different doses of hemopoietic cells containing the unique genetic markers at the expense of integration of human adenosine deaminase gene is analyzed. Low dose of donor hemopoietic cells accelerates reversion of hemopoiesis to the recipient type and decreases the number of functioning clones. This finding confirms that a stem hemopoietic cell cannot maintain itself and its proliferative potential is limited. For long and stable repopulation of the recipient subjected to gene therapy, the maximum possible number of transduced stem hemopoietic cells should be transplanted. Translated fromByulleten' Eksperimental'noi Biologii i Meditsiny, Vol. 124, No. 7, pp. 86–89, July, 1997     

Comparative study of the action of ethimizole and hydrocortisone on proliferative activity and protein synthesis in epithelial cells of the tongue and liver

Unlike hydrocortisone, ethimizole stimulated mitotic activity of the epithelial cells of the tongue and liver 6 h after its administration. The decrease in the number of mitoses in the hepatocytes after 12 h was due to the action of both substances on DNA synthesis and not to a disturbance of the entry of the cells into mitosis. Stimulation of protein synthesis was detected by autoradiographic and biochemical methods following the action of hydrocortisone and ethimizole at the maximum of inhibition of mitosis.Laboratory of Experimental Histology, Institute of Experimental Medicine, Academy of Medical Sciences of the USSR, Leningrad. (Presented by Academician of the Academy of Medical Sciences of the USSR S. V. Anichkov.) Translated from Byulleten' Éksperimental'noi Biologii i Meditsiny, Vol. 83, No. 3, pp. 348–350, March, 1977.     

Clonal succession in the hemopoietic system: Number of primitive hemopoietic stem cells and clone lifespan

The fate of individual primitive hemopoietic stem cells is studied by retroviral gene transfer technique. Tens of small hemopoietic clones with a lifespan no longer than 1 month simultaneously function throughout life in lethally irradiated mice reconstituted with bone marrow cells. The disappearing clones are not detected again, which confirms the clonal succession. The number of primitive hemopoietic stem cells in mouse bone marrow has been directly estimated: 1 per 8000 hemopoietic cells, or 30×10³ per mouse, which is at least ten times higher than expected. Translated fromByulleten' Eksperimental'noi Biologii i Meditsiny, Vol. 127, No. 4, pp. 419–423, April, 1999     

Effect of the thymus on the ability of hematopoietic stem cells to recover after sublethal radiation injury

After fractional irradiation of the bone marrow of mice (300 radin vitro + 300 radin vivo) damage to the hematopoietic stem cells is substantially less if the interval between irradiations is 5 h than if the interval is 30 min, as a result of recovery from sublethal radiation injury (the Elkind repair effect). In thymectomized adult mice ability of hematopoietic stem cells to undergo Elkind repair is severely disturbed 2.5–5 months after the operation. Transplantation of the thymus abolishes the effect of thymectomy.Laboratory of Bone Marrow Culture and Transplantation, Central Institute of Hematology and Blood Transfusion, Ministry of Health of the USSR, Moscow. (Presented by Academician of the Academy of Medical Sciences of the USSR N. A. Fedorov.) Translated from Byulleten' Éksperimental'noi Biologii i Meditsiny, Vol. 86, No. 11, pp. 584–586, November, 1978.     

Interrelationship between the membrane potential of adrenocorticocytes and functional activity of the adrenal glands in adult and old rats

Experiments on adult (5–7 months old) and old (28–30 months old) male rats reveal that ACTH induces a hyperpolarization of the plasma membrane of adrenocorticocytes from the fasciculate zone in both age groups of animals 24 hours after a single administration of the hormone. Actinomycin D, an inhibitor of protein synthesis, prevents the development of hyperpolarization. Translated fromByulleten' Eksperimental'noi Biologii i Meditsiny, Vol. 117, № 3, pp. 302–303, March, 1994 Presented by D. F. Chebotarev, Member of the Russian Academy of Medical Sciences.     

用Flu和Ara-c的改良方案预处理后自体造血干细胞移植治疗急性白血病临床观察

目的评价采用氟达拉滨（Flu）和阿糖胞苷（Ara-c）改良的预处理方案进行自体造血干细胞移植（AHSCT）治疗急性白血病的安全性和疗效。方法27例急性白血病患者,其中男性18例,女性9例;年龄12～51岁,中位年龄20岁。急性淋巴细胞白血病（ALL）21例;急性髓细胞性白血病（AML）5例。均采用包含Flu和Ara-c的预处理方案进行AHSCT,方案包括全身照射（TBI）7～8Gy或白消胺（Bu）3.2mg/（kg·d）×3d联合环磷酰胺（Cy）50mg/（kg·d）×2d、Flu30mg/（m^2·d）×3d、Ara-c2g/（m^2·d）×3d。结果除1例患者早期死亡外,所有患者均成功重建造血系统,中性粒细胞恢复至大于0.5×109/L、血小板恢复至20×109/L的中位时间分别为11（9～19）d和16（10～55）d。预处理过程中无严重不良事件发生,移植相关死亡率（TRM）7.4%（2例）。4年总体复发率（RR）、TRM和无病生存率（DFS）分别为38.8%±10.3%、9.3%±5.7%、60.8%±9.8%。结论在AHSCT治疗急性白血病中,以Flu、Ara-c改良的TBI/Cy或Bu/Cy预处理方案髓外毒性小,无病生存率较高,可作为急性白血病预处理的一种有效选择。     

Acute extinction of exploratory behavior in mice: Role of the zoosocial factor and the effects of pyracetam and scopolamine

An important role of the zoosocial factor in the acute extinction of exploratory behavior (habituation) is demonstrated by showing that habituation of mice to the new environment of a testing chamber occurs more rapidly and is more strongly marked when a group of animals is placed in the chamber than when animals are placed there singly. The nootropic drug Pyracetam, which improves cognitive functions, accelerated the habituation process only in grouped mice, whereas the amnesia-promoting drug scopolamine inhibited this process in both grouped and single mice. Translated fromByulleten' Eksperimental'noi Biologii i Meditsiny, Vol. 122, No. 7, pp. 48–50, July, 1996     

Effects of Angiotensin II and proliferative activity of epitheliocytes and smooth muscle cells of the tracheobronchial system in newborn albino rats

Angiotensin II was injected intraperitoneally in a dose of 5×10⁻⁸ mol/kg to newborn rats from then 2nd to 6th day of life. Autoradiography with³H-thymidine showed that angiotensin II stimulated DNA synthesis in epitheliocytes and smooth muscle cells of the trachea and large (cartilaginous) and small (noncartilaginous) bronchi, intensified lipid peroxidation, and activated the antioxidant defense system in the lungs.     

Effect of prenatal stress on proliferative activity and chromosome aberrations in embryo brain in rats with different excitability of the nervous system

Stress during pregnancy affects the morphogenesis of embryonal brain, its structural and functional characteristics, and behavior of the progeny. Genetic mechanisms of this process remain unclear. Cytogenetic characteristics of neuroblasts were analyzed in 17–18-day embryos of rats selected by threshold excitability of the nervous system in health and after emotional painful stress during the third trimester of pregnancy. The strains differed by the effect of stress on proliferative activity and chromosome aberrations in cells of the future hippocampus depending on the strain-specific characteristics of the nervous system excitability. This effect is regarded as an important component of epigenetic regulation of neurogenesis and behavior. Translated fromByulleten' Eksperimental'noi Biologii i Meditsiny, Vol. 129, No. 4, pp. 452–455, April, 2000     

Transformants of Neurospora crassa with the nit-4 nitrogen regulatory gene: copy number,growth rate and enzyme activity

Summary nit-4 is a pathway-specific regulatory gene which controls nitrate assimilation in Neurospora crassa, and appears to mediate nitrate induction of nitrate and nitrite reductase. The NIT4 protein consists of 1090 amino-acid residues and possesses a single GAL4-like putative DNA-binding domain plus acidic, glutaminerich, and polyglutamine regions. Several mutants with amino-acid substitutions in the putative DNA-binding domain and a nit-4 deletion mutant, which encodes a truncated NIT4 protein lacking the polyglutamine region, are functional, i.e., they are capable of transforming a nit-4 mutant strain. However, transformants obtained with most of these nit-4 mutant genes possess a markedly reduced level of nitrate reductase and grow only slowly on nitrate, emphasizing the need to examine quantitatively the affects of in vitro-manipulated genes. The possibility that some mutant genes could yield transformants only if multiple copies were integrated was examined. The presence of multiple copies of wild-type or mutant nit-4 genes did not generally lead to increased enzyme activity or growth rate, but instead frequently appeared to be detrimental to nit-4 function. A hybrid nit-4-nirA gene transforms nit-4 mutants but only allows slow growth on nitrate and has a very low level of nitrate reductase.     

Evaluating the potential impact of mismatches outside the antigen recognition site in unrelated hematopoietic stem cell transplantation: HLA-DRB1*1454 and DRB1*140101

DNA sequencing of 268 individuals drawn from four US populations carrying two unresolved DRB1*14 alleles differing only outside the antigen recognition site identified DRB1*1454 in the majority. A database of 4222 human leukocyte antigen (HLA)-matched hematopoietic stem cell transplantation donor–recipient pairs was queried to determine the number likely mismatched for DRB1*140101/DRB1*1454 but matched for class I loci. A power calculation suggests that more than 88,000 transplants among European Americans will be needed to identify sufficient 7/8 allele-matched pairs to evaluate the impact of the DRB1*140101/DRB1*1454 mismatch on transplant outcome. Molecular modeling of the HLA-DR interaction with the T-cell receptor and with CD4 suggests that the amino acid substitution distinguishing the two alleles will have minimal impact on allorecognition.     

不同预处理移植后树突状细胞早期重建及与移植物中CD34+细胞相关性分析

目的：比较不同预处理异基因造血干细胞移植（allo-HSCT）后早期树突状细胞（DCs）亚群重建情况，以及移植物中CD34^＋细胞是否影响移植后早期DCs亚群重建。方法：采用三色流式细胞仪动态检测不同预处理移植后早期外周血树突状细胞亚群DC1、DC2水平。结果：移植后早期清髓性移植患者体内DCs亚群数量非常低，常规移植组移植后14天与半相合移植组相比，DC1、DC2均无统计学意义（P〉0．05）。非清髓性移植组（NST）DC1、DC2高于清髓性移植组，两者相比具有统计学意义（P〈0．05）。在30天和60天，所有组DC1、DC2略有波动，但是幅度不大。以输入的CD34^＋细胞数平均分为三组，三组患者DC1、DC2移植后14、30和60天均无统计学意义（P〉0．05）。结论：NST后患者早期DCs重建较清髓性干细胞移植患者早，而常规移植和半相合移植早期DCs重建较慢，二者无差别。移植物中的CD34^＋细胞不影响移植后早期DCs亚群重建。     

Polymorphisms in the TNFA gene promoter region show evidence of strong linkage disequilibrium with HLA and are associated with delayed neutrophil engraftment in unrelated donor hematopoietic stem cell transplantation

Sustained myeloid engraftment is an important determinant of outcome in hematopoietic stem cell transplantation (HSCT). Human tumor necrosis factor (TNF)-alpha is encoded by a gene, TNFA, located in the class III region of the major histocompatibility complex on chromosome 6, flanked by the human leukocyte antigen (HLA) class I and II regions. A number of polymorphisms in the promoter region of the TNFA gene have been associated with increased production of TNF-alphain vivo. Additionally, raised TNF-alpha levels have been reported to have a detrimental effect on the outcome in HSCT, in particular on early complications such as acute graft vs host disease, failure to engraft, and transplant-related mortality. There is evidence of linkage disequilibrium (LD) between TNFA promoter polymorphisms and extended HLA haplotypes. We have genotyped 73 cell lines and 189 donor/recipient pairs (undergoing HSCT) for their TNFA polymorphism, all of which had been well characterized with respect to their HLA genes. We found evidence of strong LD between HLA genes and TNFA; however, there was also evidence for recombination events having taken place, as we found that a number of transplant pairs who were matched for their HLA haplotypes were not matched for their TNFA alleles. We analyzed early outcomes in the transplant recipients and found a significant delay in engraftment in those pairs where both donor and recipients possessed an AG allele (associated with higher TNF-alpha levels). Our results suggest a functional effect of TNFA polymorphisms on myeloid engraftment in unrelated HSCT.     

Analysis of FOXF1 and the FOX gene cluster in patients with VACTERL association

VACTERL association, a relatively common condition with an incidence of approximately 1 in 20,000 -35,000 births, is a non-random association of birth defects that includes vertebral defects (V), anal atresia (A), cardiac defects (C), tracheo-esophageal fistula (TE), renal anomalies (R) and limb malformations (L). Although the?etiology is unknown in the majority of patients, there is evidence that it is causally heterogeneous. Several studies have shown evidence for inheritance in VACTERL, implying a role for genetic loci. Recently, patients with component features of VACTERL and a lethal developmental pulmonary disorder, alveolar capillary dysplasia with misalignment of pulmonary veins (ACD/MPV), were found to harbor deletions or mutations affecting FOXF1 and the FOX gene cluster on chromosome 16q24. We investigated this gene through direct sequencing and high-density SNP microarray in 12 patients with VACTERL association but without ACD/MPV. Our mutational analysis of FOXF1 showed normal sequences and no genomic imbalances affecting the FOX?gene cluster on chromosome 16q24 in the studied patients. Possible explanations for these results include the etiologic and clinical heterogeneity of VACTERL association, the possibility that mutations affecting this gene may occur only in more severely affected individuals, and insufficient study sample size.