The Influence of Permafrost Microorganisms on Monocytes Differentiation In Vitro.

Metabolites of fossil microorganisms of dispersed watered rocks that have passed into a frozen state (Bacillus sp. strains 2/09 and 9/08, Bacillus megaterium 8-75) can modulate the differentiation activity of human peripheral blood monocytes into phenotypically different subpopulations when cultured in vitro for 7 days. This effect is largely determined by the type of metabolites that depends on the temperature of their production: cold (collected after bacterium culturing at 5°C), medium (at 22°C), and warm temperature (at 37°C). All three types of metabolites had a weak negative influence on the level of classical (CD14hiCD16-) monocytes and stimulated the differentiation of intermediate (CD14+CD16+) and non-classical (CD14loCD16+) monocytes. The monocytes differentiation into the subpopulation of intermediate (CD14+CD16+) was stimulated to a greater extent by medium-temperature metabolites of the strain 8/75 and into the subpopulation of non-classical (CD14loCD16+) monocytes by warm metabolites of the strains 8/75 and 2/09 throughout culturing of mononuclear cells (days 1, 3, 7). Bearing in mind the anti-infection activity of intermediate and non-classical monocytes, we can consider strains 8/75 Bacillus megaterium and 2/09 Bacillus sp. promising for their in-depth testing.

Influence of Paleobacteria on the Proliferative Activity of Human Lymphocytes In Vitro.

When testing the proliferative activity of 14 strains of permafrost microorganisms in the reaction of blast transformation of human lymphocytes in vitro, a strain (Alcaligenes sp.) with mitogen properties was isolated (20-fold increase in the rate of lymphocyte proliferation in comparison with the control). Four strains activated lymphocyte proliferation by 3-9 times in comparison with the control. Three strains produced substances with cytostatic properties and reduced proliferation activity by 33-43% and one strain (Bacillus sp.) almost completely suppressed phytohemagglutinin-induced lymphocyte proliferation. These data indicate that strains with a unique immunobiological potential are concentrated in the population of permafrost microorganisms that have undergone rigorous evolutionary selection.

Influence of the Microbiota of the Cryolithozone Paleoecosystems on the Behavioral Functions of the Nervous System of Chickens Gallus gallus.

We studied the influence of microorganisms isolated from permafrost on the psychophysiological parameters of birds. Significant effect of the microbiota of the paleoecosystems of the cryolithozone on locomotor activity, psycho-emotional state, and psychophysiological lateralization of brain function of Gallus gallus chickens. The involvement of both the autonomic and the higher central nervous systems in this regulatory process via synthesis of neuropeptides by symbionts is discussed.

[Cytogenetic, kariopathological and morphological abnormalities of spermatozoa and urothelial cells in human granulocytic anaplasmosis depending on polymorphism of the gene of DNA ligase IV].

AIM: to assess the role of human granulocytic anaplasmosis (HGA) caused by Anaplasma phagocytophilum in the induction of cytogenetic damage of spermatozoa and karyopathological abnormalities of urothelial cells depending on the polymorphism of the gene of enzyme, DNA ligase IV. MATERIAL AND METHODS: A total of 129 male patients with HGA and 84 otherwise healthy donors were examined. The samples of both semen and urothelial cells were obtained from each individuals for microscopic analysis. The diagnosis was confirmed by cytological (microscopic) method, enzyme immunoassay and polymerase chain reaction (PCR). An analysis of the frequencies of damaged spermatozoa and urothelial cells in all participants was carried out. In addition, a molecular cytogenetic study of spermatozoa by fluorescent in situ hybridization (FISH) was carried out using an AneuVysion multicolour for chromosomes 18 and 21 ("Abbott", USA) to determine the frequency of aneuploidy in spermatozoa. The level of DNA fragmentation was studied by SCD (Sperm Chromatin Dispertion Test) using a commercial Halosperm kit ("Halotech DNA", Spain). RESULTS: The cytological analysis revealed the significant increase in the proportion of spermatozoa with cytogenetic abnormalities and urothelial cells with karyopathological damage in the HGA patients. The most significant damage to nuclear structures of cells was determined in the patients with Ile/Ile genotype. The significant effects of HGA in DNA damage and cytogenetic abnormalities in patients were verified by the increased frequency of spermatozoa with DNA fragmentation, monosomy and disomy in 21 and 18 chromosomes, as well as the appearance of urothelial cells with karyopathological abnormalities. In addition, the increased frequencies of pathospermia with pathological abnormalities of head, neck and tail of spermatozoa in HGA patients were found. CONCLUSION: According to our results, the cytological analysis in the patients with HGA demonstrated the significant increase in the frequencies of spermatozoa with head, neck and tail defects and DNA fragmentation, monosomy and trisomy of the 18th and 21st chromosomes, as well as the increase in the frequencies of urothelial cells with karyopathological abnormalities. The genetic polymorphism of the effects of HGA was revealed, and the most significant cytogenetical damage was found in the patients carrying the Ile/Ile genotype of the LIG4 Thr9Ile gene.

Effect of Contamination with Perennial Permafrost Microorganisms on the Outcome of Closed Brain Neurotrauma.

We studied the effect of contamination with Bacillus genus microorganisms isolated from perennial permafrost samples on the outcome of closed brain neurotrauma in Wistar rats. It was found that contamination with different Bacillus strains produced different effects on the mortality of experimental animals with closed neurotrauma. The complex of metabolites from strain Ch2/9 - Bacillus spp. (pumilus) produced a protective effect in experimental closed brain neurotrauma.

Effects of temperature on biological activity of permafrost microorganisms.

The number and viability of microorganism specimens Bacillus spp. isolated from permafrost soil remained unchanged after incubation at temperatures of -16-37°C. Experiments on F1 CBA/Black-6 mice showed that incubation of bacteria at -5°C for 72 h promotes a decrease in their toxicity and an increase in their immunostimulating effect.

Effects of permafrost microorganisms on skin wound reparation.

Local application of ointment with Bacillus spp. strain MG8 (15,000-20,000 living bacterial cells), isolated from permafrost specimens, on the skin wound of about 60 mm(2) stimulated the reparation processes in experimental mice. A possible mechanism stimulating the regeneration of the damaged tissues under the effect of MG8 could be modulation of the immune system reactivity with more rapid switchover to humoral immunity anti-inflammatory mechanisms aimed at de novo synthesis of protein.

Experimental study of the effects of permafrost microorganisms on the morphofunctional activity of the immune system.

The immunobiological potential of a new microorganism species isolated from permafrost specimens (PMO strain 3M) collected from the Mammoth Mountain in Yakutia was studied in laboratory mice. PMO injected intraperitoneally in doses of 2500 to 50 × 10(6) microbial bodies caused characteristic dose-dependent effects on the structure and functions of the immune system (thymus and spleen indexes, functional activity of splenic macrophages, cellular and humoral immunity). Doses of PMO stimulating functional activities of both cellular and humoral immunity were detected.

[Relict microorganisms of cryolithozone as possible objects of gerontology].

Permafrost is widely distributed in the Northern Hemisphere, and its age reaches hundreds of thousands and millions of years. Permafrost contains alive microorganisms which are not frozen due to relatively high temperature of the environment (-2...-8 degrees C), but the microorganisms are immobilized and therefore aged probably similar to the age of permafrost. Longevity of the relict microbial cells is related obviously to their mechanism of protection against heat, radiation, free radicals and other damaging agents. A strain of Bacillus sp. was isolated from permafrost aged of about 3 million years, 16S rDNA sequence was identified and preliminary testing of bacterial culture on Drosophila melanogaster and mice was made. Immune stimulation and improvement of physical condition were observed, and that, together with the age of the microbial cells, presents the relict microorganisms as objects of gerontology.