Metabolic Regulation of Mammalian Stem Cell Differentiation.

Formation of normal tissue structure, homeostasis maintenance, and tissue damage repair require proliferation and differentiation of stem cells. A distinctive feature of these cells is a unique organization of metabolic pathways, in which contribution of energy production mechanisms to the general cellular metabolism is principally different from that in differentiated cells. Moreover, metabolic changes during differentiation of embryonic and postnatal stem cells have several specific features. The alterations in the stem cell metabolism are not simply consequences of cell differentiation, but also active regulators of this process. Metabolic enzymes and intermediates control and guide the maintenance of stemness, self-renewal, and differentiation of stem cells. The review discusses the patterns and molecular mechanisms of the switch in the metabolism of stem cells during their transition from the pluripotent to differentiated state with the special emphasis on how metabolic processes occurring in the stem cells regulate their functions, ability to differentiate, and the choice of the direction for development.

[Interferon-regulating activity of the CelAgrip drug and its influence on the formation of reactive oxygen species and expression of innate immunity genes in Burkitt's lymphome cells cultures.]

INTRODUCTION: Interferons (IFN) and IFN inducers are effective in suppressing viral reproduction and correcting of the innate immunity mechanisms. The aim of the study was to test the hypothesis of the possible involvement of the IFN inducer CelAgrip (CA) as an activator or suppressor of antiviral effects in Burkitt's lymphoma (LB) cell cultures with different ability to produce Epstein-Barr virus antigens (EBV). MATERIAL AND METHODS: The kinetic analysis of the dynamics of reactive oxygen species (ROS) production and determination of gene group expression by real-time PCR in response to CA treatment were done in human cell lines LB P3HR-1 and Namalva, spontaneously producing and not producing EBV antigens. RESULTS AND DISCUSSION: When treating CA in Namalva cells, a decrease in the ROS activation index was found; in P3HR-1 cells, an increase was observed. After treatment with CA, there was no reliable activation of the IFN-α, IFN-ß and IFN-λ genes in Namalva cells, but the expression of the ISG15 and P53(TP53) genes was increased more than 1200 times and 4.5 times, respectively. When processing the CA of P3HR-1 cells, the expression of IFN-α genes increased by more than 200 times, IFN-λ - 100 times, and the ISG15 gene - 2.2 times. The relationship between IFN-inducing action of CA and the activity of ISG15 and ROS in LB cell cultures producing and not producing EBV antigens is supposed. CONCLUSION: In Namalva cells that do not produce EBV antigens the treatment of CA results in suppression of ROS generation and activation of the expression of genes ISG15 and P53 (TP53); in P3HR-1 cells producing EBV antigens, the opposite picture is observed - the formation of ROS and the expression of the IFN-α and IFN-λ genes are activated and the activity of the ISG15 and P53 (TP53) genes is suppressed.

[Interferon-regulating activity of the celagrip antiviral drug and its influence on formation of reactive oxygen species and expression of innate immunity genes in the follicular lymphoma patients].

INTRODUCTION: Medicines from the group of interferon inducers (IFNs) "swith on" the synthesis of type 1 interferons (IFN-I) and induce the expression of IFN-stimulated genes (ISGs) that regulate innate immunity reactions and protect the host from infectious agents and the tumour pathology.The purpose of the study was to determine the role of the drug celagrip (CA) in the activation of innate immunity genes and the effect on the production of reactive oxygen species (ROS) in patients with follicular lymphoma (FL). OBJECTIVES:  to study the intensity of ROS production and the level of expression of the IFN-α2, IFN-λ1, ISG15, BCL2, P53(TP53) and USP18 genes in response to the treatment of blood cells of patients with FL with the preparation of CA. MATERIAL AND METHODS: The study involved primary cancer patients diagnosed with follicular lymphoma (FL) and healthy volunteers. A kinetic analysis of the dynamics of production of reactive oxygen species (ROS) was performed in whose blood cells, and the expression of the group of genes was determined by real-time PCR in response to CA processing. RESULTS AND DISCUSSION: ROS production by blood cells of patients with FL and volunteers in the presence of CA significantly decreased (P < 0.05). The level of gene expression of ISG15, P53(TR53) and USP 18 in the group of patients with FL was significantly higher than that in the group of volunteers. When treating blood cells with CA, it becomes possible to divide patients with FL into groups with a positive and negative response in accordance with the level of expression of the USP18 gene. We divided FL patients into groups with a positive and negative response in accordance with the level of USP18 gene expression after treatment of blood cells with CA. CONCLUSIONS: The CA drug reduces the production of ROS and simultaneously stimulates the activity of the innate immunity genes ISG15, P53(TP53) and USP18 in the blood cells of patients with FL.

[Cytokine-regulating activity of anti-virus preparation CelAgripus in Burkitt's lymphoma stable B-cell lines.]

INTRODUCTION: Cytokines activated in response to immunosuppressive viral infections can directly or indirectly affect the neoplastic transformation of B cells. In this study, we studied a new substance designed to produce the antiviral drug CelAgrip (CA, CelAgripus), which exhibits interferon (IFN) and cytokine-inducing activity and, apparently, can be used as an activator of antiviral immunity. Purpose - is to evaluate the cytokine-regulating effect of CA in Burkitt's lymphoma (LB) cell lines latently infected with the Epstein-Barr virus (EBV). OBJECTIVES: to study the CA-induced expression of the cytokine genes IL-1ß, IL-2, IL-4, IL-6, IL-8, IL-10, IL-12, IL-17, IL-18, IFN-α, IFN -γ, IFN-ß, IFN-λ1, IFN-λ2, IFN-λ3, TNF-α in normal and EBV transformed LB cells. MATERIAL AND METHODS: Cell line: the human embryo fibroblasts (HEF), Namalva, Daudi, Raji, P3HR-1. Preparations: CA, gossypol-acetic acid (GAA), sodium carboxymethyl cellulose (Na-CMC). METHODS: RT-PCR and methods for assessing cytotoxicity (MTT and Scepter 2.0 Merck cell counter). RESULTS: The effect of the CA preparation on the expression of IFN-λ, IL-1ß, IL-6, IL-8 and IL-10 genes was revealed. DISCUSSION: We observed the activation of gene expression of IFN-λ, IL-1ß, IL-6, IL-8 and suppression of IL-10 gene activity when treatment CA of LB cells. CONCLUSION: The substance CA has new effects on the activation of the expression of a number of key cytokine genes in stable Burkitt lymphoma cell lines.

[Ophthalmic drug films in the therapy of experimental herpetic keratoconjunctivitis.]

INTRODUCTION: The complexity of the treatment of herpetic keratoconjunctivitis is due to the severity of the disease, complications, the transition to chronic relapsing forms and the insufficient effectiveness of the drugs used, which leads to a steady increase in the number of patients. The aim of the study was to evaluate the therapeutic efficacy of the eye drug films «GlazAvir¼ in the experimental model of acute herpetic eye infection in rabbits. RESEARCH OBJECTIVES: to study the specific activity of «GlazAvir¼ and compare the long-term indicators of the level of manifestation of individual clinical signs of keratoconjunctivitis. MATERIAL AND METHODS: In the work we used rabbits of the Chinchilla breed, the herpes simplex virus type 1 and the eye drug films «GlazAvir¼. A model of ophthalmic herpetic infection was formed by infection of rabbits with virus-containing material of a pre-scarified eye cornea against the background of local anesthesia. Аnimals were treated with the drug «GlazAvir¼ - 1 application per day for 7 days. Animals were observed daily for 15 days, then every 3 days until the 25th day of observation. The effectiveness of the drug was evaluated at the peak of the development of the pathological process. RESULTS AND DISCUSSION: There was a decrease in mortality from 50 to 20%, and an increase in average life expectancy by 27.87%, compared with the control in animals treated with «GlazAvir¼. It was noted after activation of herpetic keratoconjunctivitis on the 2nd - 5th day, at the peak of the disease (6-9th day) a statistically significant decrease in the activity of the pathological process (p<0.05) by rabbits treated with the «GlazAvir¼ ophthalmic drug films. The tendency to normalization by the rabbits treated with the «GlazAvir¼ preparation was observed until the 14th day. CONCLUSION: The data obtained indicate the pronounced effectiveness of the «GlazAvir¼ preparation in the treatment of experimental herpesvirus keratoconjunctivitis.

Polyprenyl Phosphates Induce a High Humoral and Cellular Response to Immunization with Recombinant Proteins of the Replicative Complex of the Hepatitis C Virus.

The search for new adjuvants remains the critical task for the creation of hepatitis C vaccines due to the weak immunogenicity of biotechnological products. When immunizing mice with the recombinant proteins NS3 and NS5B of the hepatitis C virus (HCV), the adjuvant activity of three immunomodulators was compared. Phosprenyl® on the basis of polyprenyl phosphate (PPP), chemically synthesized analogue of the bacterial cell wall glucosaminyl muramyl dipeptide (GMDP), and IFN-α recombinant protein were tested. GMDP increased the activity of IgG1 antibodies 4-6 times but did not stimulate the production of IFN-γ; IFN-α has not shown any adjuvant properties. The introduction of recombinant HCV proteins together with PPP in low doses increased the activity of IgG2a isotype antibodies 4-7 times and increased IFN-γ secretion 3 times. Thus, it was first shown that PPP polarizes the immune response to Th1-type and is a promising adjuvant for the development of a vaccine against hepatitis C.

[Efficacy of polyprenyl phosphates in the experimental genital herpes model].

An experimental model of the primary genital herpes (herpes simplex type 2, HSV-2) in the female guinea pigs was suggested to study the infectious process activity of polyprenyl phosphates (PPP) and PPP+acyclovir (AC) complex treatment. The morphofunctional features of the guinea pig ovaries were studied in the control and experimental groups (the latter were inoculated with PPP and/or AC as a primary infection treatment) at the stage of the recurrent genital herpes aggravation. It was shown that in the case of combined PPP +AC use significant changes in the disease symptoms were observed, as well as a decrease in the infectious process activity and duration, and positive remote effect on the ovarian morphophysiology.

[Structural changes of the adrenal cortex in experimental genital herpes virus infection].

The reactive changes in the adrenal gland cortex were studied in mature female guinea pigs (n=5) in an experimental model of acute genital herpes virus infection. The methods of light and transmission electron microscopy were used. To confirm the presence of viral antigen in the corticosterocytes (CSC), the methods of immunfluorescence and electron microscopic immunocytochemistry were used. It was shown that at day 7 of an acute process, focal CSC reactive changes appeared in the glomerular zone - at the light microscopic level, CSC had intact nuclei and optically empty cytoplasm, while at the electron microscopic level, these CSC demonstrated the damaged membranous organelles, and various membranous structures which were not found in the normal cells. The aggregates of hypertrophied CSC were found in the fasciculate zone. The changes described were reversible, as they practically disappeared by the onset of spontaneous recovery (day 21 after inoculation). The regeneration of CSC of glomerular and fasciculate zones of the adrenal cortex involves both intracellular and cellular mechanisms.

[Critical conditions as a logical and appropriate series of events in iron methabolism disorders (the summary of experimental studies)].

It is revealed that in all models of critical conditions will activate the free-radical oxidation, decreasing the total antioxidant activity, the concentration of transferrin decreases in the serum of the blood, increase of the concentration of ferritin, the theological properties of the blood are violated and the signs of endothelial dysfunction are identified. Pre-entered deferoxamine in the dose of 80 mg/kg reduced the intensity of free-radical oxidation processes, restoring the antioxidant potential, concentration of the transferrin, and a lower level of ferritin, contributed to the normalization of blood theological properties and a reduction of the extent endothelium destruction as a result of the reduction Fe2+ concentration in blood serum.

Morphofunctional changes in guinea pig ovaries in experimental chronic herpesvirus infection.

The ovaries of adult guinea pigs infected with herpesvirus-2 were examined by light microscopy under conditions of latent infection and relapse of infectious process. Morphofunctional changes in the ovarian follicles at all stages of folliculogenesis and in ovarian stroma were caused by the negative effects of type 2 herpes simplex virus.

[The new chemical preparation arbidol: its prophylactic efficacy during influenza epidemics]. / Novyi khimiopreparat arbidol: profilakticheskaia éffektivnost' vo vremia épidemii grippa.

In four controlled observations on the organized groups of students and industrial workers during the epidemics of influenza A in 1988-1989 Arbidol, a new chemopreparation, was found to be well tolerated and to produce a pronounced prophylactic effect when administered in a dose of 0.2 g (2 tablets of 0.1 g) daily for 10-18 days. After the 18-day prophylactic course of Arbidol the induction of serum interferon was found.

[The antiviral activity of amiksin and its effect on the interferon status in hepatitis in mice]. / Protivovirusnaia aktivnost' amiksina i ego vliianie na interferonovyi status pri gepatitie myshei.

A single administration of amixin followed by oral infection with mouse hepatitis virus provided protection of 40%-50% animals for 72 hours and normalized the interferon status. In parallel with decreasing level of virus-induced endogenous interferon the interferon-inhibiting activity of lymphocytes of the infected animals increased and approached normal levels.

[Interferon-inducing activity of amixin and its effect on the interferon status]. / Interferonindutsiruiushchaia aktivnost' amiksina i ego vliianie na interferonovyi status.

The national preparation amixine induces interferon (IF) production in most of the people who have background values of serum IF. When the initial values of serum IF are high, after administration of amixine they decline to the background ones. A certain group of subjects (approximately 25%) is not sensitive to amixine. The IF content in the blood serum of such subjects does not change before or after administration of amixine. Its use should be limited to 4-5 administrations weekly, because its longer use leads to the exhaustion of the IF system in people.

[Influenza C: the immunological aspects of the problem]. / Gripp C: immunologicheskie aspekty problemy.

The use of the complex of methods for the characterization of cell-mediated and humoral immunity to influenza C virus has made it possible to establish that with the increase of the age of children and, simultaneously, with the increase of the number of persons found to be seropositive to influenza C the asymptomatic forms of this infection occur more frequently. Practically all examined adults selected by random choice have proved to be immune to this infective agent. The results of this investigation indicate that influenza C virus constantly circulates among the population.

[Evaluation of the interferon status of human subjects by whole-blood specimens]. / Otsenka interferonovogo statusa liudei po probam tsel'noi krovi.

A simple method for the determination of three parameters of human interferon status: serum interferon (IFN) and induction of alpha- and gamma-IFN in specimens of 10-fold-diluted whole blood in a volume of 200 microliters is described. Standard parameters of interferon status of normal subjects in the whole blood are given which can be visually presented as a triangle of interferon status, and its deviations in various pathological conditions. The whole-blood method is recommended for large-scale clinical surveys, especially in pediatrics.

[Antiviral activity of amiksin incorporated into liposomes]. / Protivovirusnaia aktivnost' amiksina, zakliuchennogo v liposomy.

The results of a comparative study of the antiviral activity of amyxin (tilorone) without liposomes and incorporated into liposomes on the model of experimental enteral hepatitis of mice are presented. The use of amyxin in liposomes was shown to prolong interferon production and to provide significant 60% protection of the animals. The possible mechanism of increasing the antiviral effectiveness of amyxin upon its incorporation into liposomes, enteral administration and subsequent enteral infection of mice with hepatitis virus is discussed. It is assumed that use of amyxin in liposomes may increase its effectiveness in prevention of human hepatitis A.