ABCA1 mRNA and protein levels in M-CSF-activated macrophages from patients with arterial stenosis

ABCA1 transporter is one of the key factors defining the level of antiatherogenic HDL in plasma. It is involved in cholesterol removal from peripheral tissues by reverse cholesterol transport. However, the influence of ABCA1 mRNA and ABCA1 protein levels in macrophages on atherosclerosis remains unexplored. Using real-time PCR, we determined the ABCA1 mRNA level in macrophages cultured for 5 days with macrophage colony-stimulating factor (M-CSF). The ABCA1 mRNA level in macrophages from patients with arterial stenosis was increased compared to the control group, p = 0.04. Western-blot assayed ABCA1 protein content in macrophages from patients was significantly lower than in the control group, p = 0.01. Our results suggest that ABCA1 mRNA and ABCA1 protein levels in macrophages may be important factors in the development of atherosclerosis.     

Specificity of action of extracellular proteases from Streptomyces spp.]

The ability of an Actinomyces strain--Streptomyces spp. to produce extracellular proteases has been studied under varied cultivation conditions during the growth cycle. The activity of enzyme preparations precipitated from the culture liquid was determined with various substrates--gelatin, casein, fibrinogen, fibrin and collagen. The isolated enzyme complex possessed caseinolytic, fibrinolytic, thrombolytic and collagenolytic activities.     

Molecular analysis of Gaucher disease: distribution of eight mutations and the complete gene deletion in 27 patients from Germany

Gaucher disease is the most common lysosomal storage disease with a high prevalence in the Ashkenazi Jewish population but it is also present in other populations. The presence of eight mutations (1226G, 1448C, IVS2+1, 84GG, 1504T, 1604T, 1342C and 1297T) and the complete deletion of the β-glucocerebrosidase gene was investigated in 25 unrelated non-Jewish patients with Gaucher’s disease in Germany. In the Jewish population, three of these mutations account for more than 90% of all mutated alleles. In addition, relatives of two patients were included in our study. Restriction fragment length polymorphism analysis and sequencing of PCR products obtained from DNA of peripheral blood leukocytes was performed for mutation analysis. Gene deletion was detected by comparison of radioactively labelled PCR fragments of both the functional β-glucocerebrosidase gene and the pseudogene. Among the unrelated patients, 50 alleles were investigated and the mutations identified in 35 alleles (70%), whereas 15 alleles (30%) remained unidentified. The most prevalent mutation in our group of patients was the 1226G (370^Asn→Ser) mutation, accounting for 18 alleles (36%), followed by the 1448C (444^Leu→Pro) mutation, that was found in 12 alleles (24%). A complete gene deletion was present in two alleles (4%). The IVS1+2 (splicing mutation), the 1504T (463^Arg→Cys) as well as the 1342C (409^Asp→His) mutations were each present in one allele (2%). None of the alleles carried the 84GG (frameshift), 1604A (496^Arg→His) or the 1297T (394^Val→Leu) mutation. This distribution is different from the Ashkenazi Jewish population but is similar to other Caucasian groups like the Spanish and Portuguese populations. Our results confirm the variability of mutation patterns in Gaucher patients of different ethnic origin. All patients were divided into nine groups according to their genotype and their clinical status was related to the individual genotype. Genotype/phenotype characteristics of the 1226G, 1448C, and 1342C mutations of previous studies were confirmed by our results. Received: 19 November 1996 / Revised: 29 January 1997     

Expression of ergotope-associated markers of T lymphocytes in atopic dermatitis after in vitro polyclonal activation

The antiergotypic response leads to the formation of effector T cells able to eliminate activated lymphocytes independently of their antigenic specificity, since the targets of these cells are molecules produced during cell activation (ergotopes). In this paper, we describe the level of expression of the ergotope-associated markers CD25, HSP60, and HLA-DR by the T lymphocytes isolated from the blood of atopic dermatitis patients immediately after isolation and after cultivation. After 10-day cultivation in the presence of anti-CD3 antibodies and IL-2, the expression levels of early and late activation markers in T cells have changed: the shares of CD25-positive CD4⁺ and CD8⁺ lymphocytes increase to 68 and 47%, respectively, and the share of HLA-DR-positive cells increases to 26 and 33%. The density of HLA-DR molecules on the surface of activated T cells increases more than fivefold. Almost all T cells before and after cultivation express 60 kDa heatshock protein (HSP60); however, the CD4⁺ cells activated in vitro contain more HSP60 molecules than do the in vitro-activated CD8⁺ cells and the CD4⁺ cells of peripheral blood. Thus, the T cells of atopic-dermatitis patients have the status of activated cells because they express sufficient amounts of early and late activation markers; presumably, they can enhance the induction of antiergotypic response when administered to patients. Taking into account that antiergotypic regulation acts on activated T cells independently of their antigenic specificity, immunotherapy utilizing autologous activated T lymphocytes can be of interest as a method for targeted action on pathogenetic components of atopic dermatitis.     

Standards for laboratory diagnostics of legionellosis and their application during epidemic outbreak of pneumonia in town Verkhnyaya Pyshma

High effectiveness of application of international standards for legionellosis laboratory diagnostics was confirmed during investigation of pneumonia outbreak in town Verkhnyaya Pyshma. Use of immunochromatographic method and enzyme immunoassay for detection of Legionella antigen in urine of patients allows to confirm diagnosis of Legionella infection during several hours, promptly begin etiologic antibacterial treatment and reveal possible sources of infection in potentially dangerous environmental objects.