Hypoxia as a Factor Involved in the Regulation of the apoA-1, ABCA1, and Complement C3 Gene Expression in Human Macrophages.

Hypoxia plays a critical role in progression of atherosclerosis. Local oxygen deficiency in a plaque creates a specific microenvironment that alters the transcriptome of resident cells, particularly of macrophages. Reverse cholesterol transport from plaque to liver is considered a main mechanism for regression of atherosclerosis. Ubiquitously expressed ATP-binding cassette transporter A1 (ABCA1) and liver- and small intestine-derived apolipoprotein A-1 (ApoA-1) are two main actors in this process. We recently reported endogenous apoA-1 expression in human macrophages. While ABCA1 and ApoA-1 have antiatherogenic properties, the role of complement factor C3 is controversial. Plasma C3 level positively correlates with the risk of cardiovascular diseases. On the other hand, C3 gene knockout in a murine atherosclerosis model increases both plaque size and triglycerides level in blood. In the present study, we show for the first time that a hypoxia-mimicking agent, CoCl2, induces the upregulation of the apoA-1 and C3 genes and the accumulation of intracellular and membrane protein ApoA-1 in THP-1 macrophages. The MEK1/2-Erk1/2 and MKK4/7-JNK1/2/3 cascades are involved in upregulation of ABCA1 and C3 via activation of transcription factor NF-κB, which interacts with the HIF-1α subunit of hypoxia-inducible factor 1 (HIF-1). The three major MAP-kinase cascades (Erk1/2, JNK1/2/3, and p38) and the NF-κB transcription factor are involved in the hypoxia-induced expression of the apoA-1 gene in THP-1 macrophages.

[Features of population composition of peripheral blood CXCR3-positive lymphocytes in chronic viral hepatitis c patients].

AIM: Quantitative determination ofCXCR3+, CCR5+ and CCR6+ cells in major lymphocyte populations: T-helpers (Th), cytotoxic T-lymphocytes (CTL), natural killers (NK) and T-natural killer cells (TNK), B-lymphocytes in patients with chronic viral hepatitis C (CVHC). MATERIALS AND METHODS: Content of lymphocyte populations carrying chemokine receptor CXCR3 was studied, chemokine receptors CCR5 and CCR6 were evaluated on T-lymphocytes, in peripheral blood of 19 CVHC patients and 32 conditionally healthy donors. Cell populations were determined by flow cytofluorometry by using various combinations of monoclonal antibodies: for evaluation of Th and CTL (CD3/CD4/CD8/CXCR3/CCR5/CCR6); NK and TNK (CDl6/CD56/CD3/ CXCR3); B-cells (CD 19/CD45/CXCR3). RESULTS: In patients with CVHV compared with healthy donors a significant increase of quantity of CXCR3-positive Th was detected, however the content of CXCR3-positive CTL did not differ in the groups compared; CXCR3+ NK cell content was lower with equal content of CXCR3+ TNK. Analysis of quantity of CXCR3+ B-cells showed an increase of more than 3.5 times in CVHC patients. Significant differences in relative content of Th and CTL carrying CCR5 and CCR6 were not detected despite a non-significant increase of quantity of CCR5+ and CCR6+ Th. CONCLUSION: Content of major lymphocyte populations carrying chemokine receptor CXCR3 changed significantly compared with conditionally healthy donors in peripheral blood of CVHC patients. The increase of quantity of CXCR3-positive B-cells may be associated with infection of these cells by HCV or development of extra-liver manifestations of HVHC.

[Expression of the starfish complement component C3 gene homologue under the influence of bacterial lipopolysaccharide].

The fragment of a homologue of complement component C3 gene has been cloned and sequenced from the starfish, Asterias rubens. Phylogenetic analysis of ArC3-like gene demonstrates that ArC3-like gene has close similarity to C3 gene homologues of Deuterostomia invertebrate animals. High level of ArC3-like gene expression was identified in circulating cells (coelomocytes), in a gut's derivate (hepatopancreas) and in male gonada but not in stomach, female gonad and rectal gland of A. rubens starfish. ArC3-like gene expression was shown in all types of starfish coelomocytes: in lymphocyte-like cells, granular and nongranular amebocytes. Injection of bacterial lipopolysaccharide (LPS) solution into the coelomic cavity of starfish leads to the increase of ArC3-like gene expression in coelomocytes and hepatopancreas over the control level of sterile sea water injection. The level of ArC3-like gene expression increased in response to LPS reaching the maximum 6 h after the stimulation, and decreased to basal level 24 h after the stimulation. Injection of LPS solution stimulated the increase of ArC3-like gene expression level in hepatopancreas reaching the maximum 6-12 h after the stimulation, and the level of mRNA of ArC3-like gene had still been increased 48 h after LPS injection. The data demonstrates sustained positive regulation of ArC3-like gene expression under the influence of LPS.

[Comparative immunological analysis of echinoderm cellular and humoral defense factors]. / Sravnitel'no-immunologicheskii analiz kletochnykh i gumoral'nykh zashchitnykh faktorov iglokozhikh.

Coelomocyte are found in the fluid filling coelomic cavity of echinoderms and depending on species can be a mixture of several morphologically different types. There are among them: granular and agranular amoebocytes, morula cells, vibratile and lymphocyte-like cells. All these cells take part in cellular response to immune challenges through phagocytosis, clotting, encapsulation of foreign particles, cytotoxicity, and the production of antimicrobial agents, such as reactive oxygen and nitric oxide. The data are given on a variety of humoral factors found in the coelomic fluid, including different types of lectines, agglutinins, hemolysins, acute phase proteins and antimicrobial factors. The discussion on cooperation between cellular and humoral arms of defense reactions during inflammation reveals the crucial role of coelomocytes in immune response. It is suggested that the sea urchin complement system (that is homologous to the alternative pathway in vertebrates) is appeared initially in echinoderms as a protein cascade that points to opsonization of foreign cells and particles, augmenting their phagocytosis and subsequent destruction by coelomocytes. So the identification of a simple complement system as a part of the echinoderm immune response shows that these animals as well as all invertebrate deuterostomes share innate immune system homologies with vertebrates. Studying the simpler immune response demonstrated by echinoderms is important for understanding the ancestral deuterostome defense system and reconstructing the evolution of immune system in higher vertebrates.

[Genetic characteristics and prognosis of the existence of the European bison free-living population created in the "Orlovsky Poles'e" National Park]. / Geneticheskie kharakteristiki i prognoz sushchestvovaniia vol'nozhivushchei populiatsii evropeiskogo zubra, sozdavaemoi v natsional'nom parke "Orlovskoe poles'e".

We present preliminary data on genetic aspects of preserving viability of the free-living population of European bison (Bison Bonasus L.) created in the Orlovskoe Poles'e National Park.

[Genealogic analysis of hereditary components of a metapopulation of Przhevalsky horse]. / Genealogicheskii analiz nasledstvennykh komponentov megapopuliatsii loshchadi przheval'skogo.

The current condition of the megapopulation of the Przhevalsky horse was assessed using genetic indices of biological diversity of species and genealogical analysis and taking into account both nuclear and non-nuclear (mitochondrial), maternally inherited components of hereditary information.

[Genetic control of spermatogenesis and sex determination in mammals]. / Geneticheskii kontrol' spermatogeneza i determinatsii pola u mlekopitaiushchikh.

The material was analyzed on the main problems of genetics of mammalian spermatogenesis, sex determination, its reversion and other defects from the standpoint of current cytological and molecular-genetic concepts of functional activity of the parental genomes after fertilization and behavior of their chromosomes at the early embroyonic stages. On the basis of this analysis, a hypothesis has been proposed, which explains a high percentage (50% or more) of early embryonic mortality in placental mammals under the conditions of natural and extracorporeal fertilization, as well as regular appearance of defects in the course of natural sex determination, including the appearance of representatives of both sex minorities. We do not make pretense to comprehensive and deep analysis of male gametogenesis and sex determination in mammals.

[Sterility of males determined by functional features of the mouse spermatozoa bearing t-complex]. / Steril'nost' samtsov, obuslovlennaia funktsional'nymi osobennostiami spermatozoidov myshei, nesushchikh t-kompleks.

The mechanisms underlying normal spermatogenesis and its pathology expressed as male sterility determined by t-complex located on chromosome 17 in mice are considered in this review. t-Complex is a very convenient model with diverse markers of expression of the genes involved in development of the functional features of the spermatozoa bearing t-complex. These features include defects of mobility, capacitation, and acrosome reactions, which determine full or partial male sterility. It has been proposed that the defects of capacitation are also inherent in humans and affect male fertility. This homology is confirmed by the presence of the male gene Tcp11 in humans and demonstration of the fact that the protein TCP11 plays a leading role in modulation of the capacitation of murine spermatozoa. Hence it follows that the defects of human genes leading to incomplete binding of the fertilization promoting peptide could play a certain role in a decreased male fertility. All this is essential not only for deeper understanding of the biology of spermatozoa, but also for development of new therapeutic methods of finding and treating the semen pathology.

[Transmission ratio disruption, sterility, and control of t-complex function in sperm]. / Narushenie sootnosheniia peredachi, steril'nost' i kontrol' funktsii spermatozoidov t-kompleksa.

Mouse t-complex located on chromosome 17 contains genes affecting solely male fertility. Some genes of this complex are recessive lethals; nonetheless, the high frequency of the t-complex carriers in a population is maintained due to a mechanism referred to as transmission ratio distortion (TRD), i.e., after crosses with wild-type females, males heterozygous for the t-complex transmit the t-bearing chromosome to nearly all their offspring, which suggests that the t-complex genes control sperm function. Analysis of this phenomenon shows that the resultant TRD is determined by the ratio between the distorter genes (Tcd) and a responder gene (Tcr) located within the t-complex region. Many authors believe that two to six distorter genes currently known have an additive effect. A genetic model of the non-Mendelian inheritance in the progeny of heterozygous male mice specifically explains sterility of animals carrying the t-complex with complementary lethal genes. The model suggests that some distorter gene products interacting with the responder gene have a selective effect on motility of both mutant and wild-type sperm. Insufficient sperm motility and/or their unsuccessful capacitation result in poor if any fertilization. Information on the t-complex genes is necessary for understanding the biological mechanisms of male sterility and may be used in medical practice.

[DNA fingerprinting and genetic diversity of the European bison (Bison bonasus), American bison (Bison bison), and gray Ukrainian cattle (Bos taurus)]. / DNA-fingerprinting i geneticheskoe raznoobrazie evropeiskogo zubra (Bison bonasus), amerikanskogo bizona (Bison bison) i serogo ukrainskogo skota (Bos taurus).

To describe genetic variability and population diversity in domesticated populations of American bison (Bison bison), aurochs (Bison bonasus), and gray Ukrainian cattle (Bos taurus) different variants of DNA fingerprinting technique (utilizing the M13 phage DNA, (TTAGGG)4 synthetic oligonucleotide, and three arbitrary primers as hybridization probes) were used. Several parameters characterizing polymorphism and genetic diversity levels in each population (species) were evaluated on the basis of the profiles obtained. Dendrograms reflecting similarities between individual animals were constructed. Genetic variability of minisatellite and telomeric markers observed in the gray Ukrainian cattle flock was higher than that in aurochs and bisons. Comparison of the intrapopulation similarity (S) and gene diversity (H) indices along with the analysis of clusters in the dendrograms showed that the relatedness between the aurochs individuals was much higher than between the individual animals in the bison and gray Ukrainian cattle flocks. Furthermore, the gray Ukrainian cattle flock was represented by more distant relatives than the bison flock. It is suggested that reduced genetic variability and the appearance of deviant genotype observed in the two bison lines under selection, resulted from close inbreeding and the founder effect. The diagnostic value and efficacy of utilization of different molecular markers for estimation of genetic diversity and relatedness in domesticated animal populations is discussed.

[Genetic diversity of the European bison herd from the Prioksko-Terrasny nature reserve]. / Geneticheskoe raznoobrazie v plemennom stade zubrov Prioksko-Terrasnogo zapovednika.

Maintaining the existing genetic diversity in populations is one of the most important measures for species conservation. Some components of this diversity in the bison population of the Prioksko-Terrasny Nature Reserve, including the level of preservation of the founder genetic diversity or the founder genomic equivalent (fge) of the current bison stock and the indices of similarity (mk) and genome uniqueness (gu), as well as their effects on viability of the animals from 1950-1995 were studied. The data obtained were treated and the diversity parameters were calculated with the use of the SPARKS software package. This was performed in collaboration with the Institute for Zoo Biology and Wildlife Research (Berlin, Germany). The results of the analysis indicated that, in breeding groups of the European bison, the fge, mk, and gu values should be maintained at levels higher than 1.30, lower than 40%, and higher than 10%, respectively. When selecting males for specific groups of females, the male gu should be at least 20%, and mk should be lower than 40%.

[Use of taxonomic typing of DNA for analyzing genomic variability in representatives of Bovidae Gray, 1821]. / Ispol'zovanie taksonomicheskogo tipirovaniia DNK dlia analiza genomnoi variabel'nosti predstavitelei Bovidae Gray, 1821.

Genome variability in representatives of Bison and Bos was studied using taxonomic DNA typing. It was shown that this method, while not revealing individual genomic differences, can be used to differentiate Bovidae representatives at the genus levels. The species-specific DNA fragments found can serve as molecular genetic markers that characterize the genetic variability of the studied animal groups at the taxonomic level.