Subpopulation Heterogeneity of NK Cells during the Genetic Modification for Subsequent Use in Targeted Therapy.

Obtaining genetically engineered NK cells is a developing area of immunotherapy. In this work, we analyzed the subset heterogeneity of NK cells subjected to retroviral transduction, taking into account the content of adaptive NK cell progenitors. It was shown that subsets KIR2DL2/DL3+, as well as CD57-KIR2DL2/DL3+NKG2C+, can be modified with greater efficiency than the corresponding subsets that do not carry the KIR2DL2/DL3 and NKG2C markers. After genetic modification, the CD57-KIR2DL2/DL3+NKG2C+ cells began to express CD57 de novo, acquiring the adaptive NK cell phenotype.

CD5+CD23+ leukemic cell populations in TCL1 transgenic mice show significantly increased proliferation and Akt phosphorylation.

B-cell chronic lymphocytic leukemia (B-CLL) is the most common adult leukemia. Deregulation of the T-cell leukemia/lymphoma 1 (TCL1) oncogene in mouse B cells causes a CD5-positive leukemia similar to aggressive human B-CLLs. We recently reported that levels of TCL1 expression in B-CLL are regulated by miR-29 and miR-181 that target 3' UTR of TCL1. To determine whether treatment with microRNAs targeting TCL1 can inhibit B-CLL in mice, we generated TCL1 transgenic mice using a construct containing the 3' and 5' UTRs of TCL1 under B-cell-specific Emicro promoter (Emicro-TCL1FL). At the age of 16-20 months, these mice showed B-CLL-like disease. Immunophenotyping revealed accumulation of CD5+CD23+B220+ population in spleens and lymph nodes. Our results show that CD5+CD23+ B-cell populations from Emicro-TCL1FL mice actively proliferate and show significantly increased levels of phospho-Akt. Emicro-TCL1FL mice showed immunological abnormalities similar to human B-CLL, including hypoimmunoglobulinemia, abnormal levels of cytokines and impaired immune response. These findings revealed biochemical and immunological similarities between Tcl1-driven B-CLL in mice and human B-CLL.

[Pharmacoprophylaxis of sea-sickness syndrome with the help of homeopathic drug "Avia-more"].

Anti-sea-sickness effect of a new homeopathic drug [see text] and its influence on operator's working capacity during Coriolis acceleration was evaluated in blind placebo-controlled investigation conducted in practically healthy volunteers with low vestibular resistance. For comparison "Dimenhydrinate" was used. The investigation has demonstrated that [see text] has rather strong anti-sea-sickness effect, is comparable with dimenhydrinate effectiveness and prevents the development of disorders in sensory-and-motor and cognitive functions during the influence of static-and- kinetic loads. According to the data obtained the protective action of [see text] is most likely connected with adaptive effect on organs and systems that form the functional protective system against sea-sickness. The negative influence of the drug on CNS complex functions was not observed that allows recommending its prophylactic use in subjects with low sea-sickness resistance including the operators.

Akt phosphorylates and regulates the orphan nuclear receptor Nur77.

The immediate early gene NUR77 (also called NGFI-B) is required for T cell antigen receptor-mediated cell death and is induced to very high levels in immature thymocytes and T cell hybridomas undergoing apoptosis. The Akt (PKB) kinase is a key player in transduction of anti-apoptotic and proliferative signals in T cells. Because Nur77 has a putative Akt phosphorylation site at Ser-350, and phosphorylation of this residue is critical for the transactivation activity of Nur77, we investigated whether Akt regulates Nur77. Coimmunoprecipitation experiments showed the detection of Nur77 in Akt immune complexes, suggesting that Nur77 and Akt physically interact. We further show that Akt specifically phosphorylates Ser-350 of the Nur77 protein within its DNA-binding domain in vitro and in vivo in 293 and NIH 3T3 cells. Because phosphorylation of Ser-350 of Nur77 is critical for its function as a transcription factor, we examined the effect of Akt on this function. By using luciferase assay experiments, we showed that phosphorylation of Nur77 by Akt decreased the transcriptional activity of Nur77 by 50--85%. Thus, we show that Akt interacts with Nur77 and inactivates Nur77 by phosphorylation at Ser-350 in a phosphatidylinositol 3-kinase-dependent manner, connecting the phosphatidylinositol 3-kinase-dependent Akt pathway and a nuclear receptor pathway.

The chum salmon insulin-like growth factor II promoter requires Sp1 for its activation by C/EBPbeta.

The chum salmon insulin-like growth factor II (IGF-II) gene is highly expressed in liver tissue. In this study we demonstrate that two transcription factors, Sp1 and C/EBPbeta, are involved in the enhanced expression of the salmon IGF-II gene. The presence of the fish homolog for C/EBPbeta in salmon liver RNA was confirmed by Northern blotting. The sIGF-II promoter was activated up to 20-fold by co-transfection with C/EBPbeta. The functional importance of four out of the five putative C/EBPbeta binding sites was demonstrated with mutational analysis in transient transfection assays. The transcription factor Sp1 binds to two sites within the salmon IGF-II promoter. Interestingly, mutation of the Sp1 binding sites decreases not only the basal IGF-II promoter activity but also the C/EBPbeta-induced transactivation. These results demonstrate that liver-enriched C/EBPbeta and ubiquitously expressed Sp1 each activate the sIGF-II promoter and that Sp1 is required for full transactivation of the sIGF-II gene by C/EBPbeta. This suggests that C/EBPbeta and Sp1 act in synergy.

The chum salmon IGF-II gene promoter is activated by hepatocyte nuclear factor 3beta.

IGF-II plays an important role in growth and development of vertebrates and is highly expressed in adult salmon liver. In the present study, we demonstrate that a liver-enriched transcription factor, hepatocyte nuclear factor 3beta (HNF-3beta), is an activator of the chum salmon IGF-II gene. Multiple binding sites for HNF-3beta were identified within the 5'-UTR using electrophoretic mobility shift assays and mutation of these sites prevents binding of HNF-3beta. In transient transfection assays it was shown that mutation of the HNF-3beta binding sites results in a substantial decrease of HNF-3beta-activated salmon IGF-II gene expression. This is the first identified transcription factor that is functionally involved in the regulation of fish IGF-II expression.

Organization and expression of the chum salmon insulin-like growth factor II gene.

IGF-II plays an important role in growth and development of vertebrates. In the present study, the characterization of the first fish IGF-II gene, chum salmon IGF-II, is described. The sIGF-II gene consists of four exons, spanning a region of 9 kbp, that encode the 214 aa IGF-II precursor. While the amino acid sequences of fully processed IGF-II of salmon and mammalian species are very similar, the prepro-peptide sequence deviates extensively in the signal- and E-peptide domains. The transcription initiation site of the sIGF-II gene was localized within a 30 nt region employing RT-PCR. Using sIGF-II promoter-luciferase constructs it was demonstrated that the sIGF-II gene has a relatively strong promoter that contains tissue-specific regulatory elements.

[Antioxidant system disorder in diabetic patients and the means for its normalization]. / Porushennia antyoksydantnoï systemy u khvorykh na tsukrovnyi diabet i shliakhy ioho normalizatsiï.

The condition was studied on the antioxidant system (AOS)--the activities of catalase, concentration of tocoferoli and reduced glutathione--in 112 patients with type I diabetes mellitus during the course of treatment. It has been established that in decompensation of diabetes mellitus there is a striking deterioration of AOS the degree of which depends upon duration and severity of the illness as well as the presence of angiopathies. Employment in the combined treatment of the donator of sulphhydric groups unithiol and a naturally occurring antioxidant tocopheroli promoted normalization of parameters associated with AOS, diabetic angiopathies included.

[The immunoglobulin content of the blood serum in patients with Addison's disease]. / Soderzhanie immunoglobulinov v syvorotke krovi bol'nykh bolezn'iu Addisona.

The authors examined 24 patients suffering of primary chronic insufficiency of the adrenal cortex, in particular, the content of immunoglobulins of A, M, G classes. Results revealed an increase of the level of immunoglobulins G in severe and average-severe forms and an increase of immunoglobulin M in severe forms of the disease.

[The motor activity of the gastrointestinal tract in acute appendicitis]. / Motornaia deiatel'nost' zheludochno-kishechnogo trakta pri ostrom appenditsite.

From study of electrogastrograms in 92 patients with acute appendicitis before and in various periods after appendectomy conducted under local or halothane nitrous-oxide-oxygen anesthesia, the authors conclude that electrogastrography may be used in complex with other methods for prognosticating the possibility of the occurrence of postoperative paresis. They point to the preventive significance of general anesthesia in the development of postoperative pareses. Percutaneous electrostimulation had a favourable effect in 12 patients with paresis developing after appendectomy.

[Assessment of the working capacity of patients with chronic cortico-adrenal insufficiency]. / Otsenka fizicheskoi rabotosposobnosti bol'nykh s khronicheskoi nedostatochnost'iu kory nadpochechnikov.

A study was made of CVS function in 48 patients with Addison's disease by submaximum bicycle ergometry to specify and improve the existing criteria of assessment of a degree of gravity of disease and the working capacity of patients before and after hormonal substitution therapy. According to the results of bicycle ergometry, the working capacity of these patients was lowered, the total volume of work done, loading power, and maximum oxygen consumption were also lowered. The data obtained can be used for assessment of therapeutic efficacy and in medical labor examination to determine the working capacity of patients with hypocorticism.