Differential Expression of Bipotent Commitment-Related Genes in Multipotent Mesenchymal Stromal Cells at Different O2 Levels.

The transcriptomic profile associated with osteo- and adipogenic differentiation in growth-arrested multipotent mesenchymal stromal cells (MSCs) from human adipose tissue was analyzed in vitro at 20% (standard laboratory) and 5% (tissue-related) O2 levels. Compared with day 7, at 5% O2 on day 14 spontaneous upregulation of osteo- (RUNX2, SP7, BGLAP, and SPP1) and adipogenic differentiation (CEBPA, PPARG, and ADIPOQ) genes in MSCs was observed (p < 0.05). Thus, upon expansion under tissue-related O2, MSCs demonstrated a bipotent transcriptomic profile, which may contribute to the improvement of their hematopoiesis-supportive function.

Analysis of the levels of tumour necrosis factor (TNF), autoantibodies to TNF, and soluble TNF receptors in patients with rheumatoid arthritis.

OBJECTIVES: To evaluate the potential contribution made by autoantibodies against tumour necrosis factor (TNF) to the pathogenesis of rheumatoid arthritis (RA). METHOD: We used affinity chromatography methods and a magnetic separation procedure to purify human autoantibodies specific to TNF. The autoantibodies were used as calibration material to determine the absolute content of autoantibodies to TNF using an enzyme-linked immunosorbent assay (ELISA). TNF content and the levels of soluble TNF receptors types I and II (sTNF-RI and sTNF-RII) were determined using commercial ELISA test kits. RESULTS: We demonstrated significant increases in the levels of TNF, sTNF-RI, and sTNF-RII in the sera of patients with acute RA and in patients with RA who had responded positively to therapy compared with healthy controls. Levels of autoantibodies of the immunoglobulin (Ig)G2, IgG3, and IgG4 subclasses were significantly higher in sera from patients with acute RA than in sera from healthy controls. Level of autoantibodies of the IgG2 subclass were significantly higher in sera from patients with acute RA than in RA patients who had responded positively to therapy. CONCLUSIONS: Acute RA is associated with changes in levels of TNF and soluble receptors for TNF and also in levels of autoantibodies to TNF. Given the magnitude of the changes in levels of different subclasses of autoantibodies to TNF, we propose that these autoantibodies might contribute to the pathogenesis of RA.

Purification of human immunoglobulin G autoantibodies to tumor necrosis factor using affinity chromatography and magnetic separation.

Autoantibodies to cytokines are important biological effector molecules that can regulate cytokine activities. The aim of the study was to develop a protocol to purify autoantibodies to tumor necrosis factor from human serum, for use as a calibration material to determine the absolute content of autoantibodies to tumor necrosis factor by enzyme-linked immunosorbent assay. The proposed protocol includes a set of affinity chromatography methods, namely, Bio-Gel P6DG sorbent to remove albumin from serum, Protein G Sepharose 4 Fast Flow to obtain a total immunoglobulin G fraction of serum immunoglobulins, and Affi-Gel 15 to obtain specifically antibodies to tumor necrosis factor. The addition of a magnetic separation procedure to the protocol eliminated contaminant tumor necrosis factor from the fraction of autoantibodies to tumor necrosis factor. The protocol generated a pure fraction of autoantibodies to tumor necrosis factor, and enabled us to determine the absolute concentrations of different subclasses of immunoglobulin G autoantibodies to tumor necrosis factor in apparently healthy donors.

[Seasonal dynamics of the structure of component communities of parasites in the young minnow Phoxinus phoxinus (L.)].

Component communities of parasites of Phoxinus phoxinus from the Chovju River near Syktyvkar City (Komi Republic) are investigated in seasonal aspect. Material for the study has been collected in May-September 2000 from 90 host specimens of ages 0+-1+. Calculations of diversity indices and other coefficients have been made by two ways -- for metazoan parasites only and for whole parasite community including protozoan species. In both these cases three states of the community have been established, 1) forming community in May-June, 2) completely formed community in the end of June-July, 3) disintegrating community in August, 4) forming community in September. Seasonal dynamics of the component communities in young minnow thus found to be different from that in adult fish hosts.

[Component parasite communities in the minnow Phoxinus phoxinus (L.) from ecologically safe and polluted reservoirs].

A comparative analysis of the structure of component parasite communities in the minnow from ecologically safe or polluted to variable extent reservoirs was made. The analysis showed that the increase of the reservoir pollution by everyday drains, washings of fertilizers from the fields around and the dacha sections causes the change of dominant parasite species in the minnow, alteration of the sum of errors in the regression equation characterizing the spread of values of biomasses of the species forming the parasite community, and change of the "graphic" structure of the community. It is proved, that the high concentration of biogens has a destructive effect on natural systems, and at the same time causes the process of self-organizing, leading to the alteration of the community structure. As soon as even a small part of biogens is stopped to come into the reservoir, and the quality of environment is restored, the community restores its structure.

[Seasonal dynamics of the component community structure of parasites of the minnow Phoxinus phoxinus (L.)]. / Sezonnaia dinamika struktury komponentnykh soobshchestv parazitov gol'iana rechnogo Phoxinus phoxinus (L).

The material represented by 75 specimens of minnow of the age 2-2(+) was collected according to the standard technique in the Chovju River (tributary of the Vychegda River, a region of the settlement Nizniy Chov, district of Syktyvkar town) during the period June-September 2000. The quantitative estimation of the structure of the component parasite communities was performed by calculating errors of the equation of regression for each species group separately, with subsequent summarizing of means of errors by all parasite groups comprising the community (Dorovskikh, 2001 6; 2002 B). In order to have a possibility to get data complementing each other, the calculation of variety indexes and other indexes have been made for metazoan parasites only and for the whole community, including the protozoan parasites. In both cases, i.e. considering only metazoan parasites and the whole composition of parasite species, three states of the component parasite community have been recognised: the formed community (June), the community in destroying (July and August), the community in the process of formation (September). In the course of working on the total species composition we recorded the beginning of community destroying on 30th of June, and the beginning of community formation in August. Considering only the metazoan parasites, the community is defined (after: Pugachev, 1999) as the mature (balanced) one in June, while in July, August and September, it was unmature (off-balance) by its characteristics. Regarding the protozoan parasites, the community in June, August and September was characterized as the mature one based on indices of parasite biomass, and as unripe one based on the number of parasite individuals; however in July, both groups of indices allowed to refer it to the unripe state. However, these unripe states are essentially different. In July, it is the result of dieing out the parasites of the past generation; in August and September, it is the result of the appearances of new generations. Therefore we recognise three states of parasite community named above. Considering the whole species composition of parasites we noted the greater difference of index values based on the parasite specimen numbers and their conventional biomass, that was in the case of the metazoan parasites only. It is particularly noticeable in the middle of the June, in the period of the formed community. This fact, together with high errors of the equations of regression and the presence of the high number of Apiosoma, points to the disturbance in the structure of component parasite community in the minnow from the Chovju River. This is easily explicable, because the Chovju River is the polluted reservoir, and pollution comes from agricultural fields, Verhny-Chov settlement, pigsties and cow-sheds. The pollution is a seasonal factor here. The most powerful pollution was noted at May-June, and then it decreased along the beginning of rains in the end of August; in September it increased again. It is important to point out that the monitoring of the metazoan parasites only allows to reveal the general dynamics of the community during the period of observation, but does not allow to catch the beginning of its destroying and developing and to notice possible disturbances in the community structure caused by pollution of reservoirs, particularly, if this pollution is a seasonal factor as in the Chovju River. Three named states of the component community of the fish parasites take place in other periods of year than this observed in the intestional parasite communities of fish helminths of the temperate climate zone. The developing of parasite communities of the intestinal helminths of the Anguilla anguilla in England (Kennedy, 1997) and of Leuciscus idus from the Rybinsk reservoir (Zhohov, 2003) starts in the beginning of summer. In May, their species diversity is minimal and in August is maximal. In conditions of the middle stream of the Vychegda River, the species diversity of parasite community associated with the minnow is maximal in June and minimal in August, when it only begins developing.

[The natural killer and fungicidal activity of the peripheral blood mononuclear cells in candidiasis]. / Estestvennaia i fungitsidnaia aktivnost' mononuklearov perifericheskoi krovi pri kandidoze.

The activity of natural killers and the capacity of peripheral blood mononuclears for the extracellular killing of Candida cells in patients with different forms of candidiasis has been determined in the radiometric test with the use of targets labeled with 3H-uridine RNAase and in the fungicidal test. On the basis of the data thus obtained the conclusion has been made that in candidiasis changes in the activity of natural killers and fungicidal capacity depend on the severity of the disease and the stage of the process. Linear correlation between the activity of natural killers and fungicidal capacity in candidiasis patients at the stage of exacerbation and in healthy persons is absent.

[Comparative analysis of methods of evaluating antibody-dependent cell-mediated cytotoxicity]. / Sravnitel'nyi analiz metodov otsenki antitelozavisimoi kletochno-oposredovannoi tsitotoksichnosti.

Two methods for measuring the activity of antibody-dependent cell-mediated cytotoxicity are compared. One of them is based on the determination of the relative amount of hemoglobin released from lysed sheep target red cells into the incubation medium via benzidine hydrochloride, that joins hemoglobin in the presence of H2O2 and stains the solution blue, the intensity of staining being measured spectrophotometrically. The authors consider radiometry preferable; this method makes use of 3H uridine-labeled chicken nuclear red cells as targets, and the results of the test are analyzed ina beta-scintillator. Less patient's blood is necessary for this test, since lower concentrations of effector cells may be used.

[Immune system function in candidiasis patients]. / Sostoianie immunnoi sistemy u bol'nykh kandidiozom.

To compare the clinical picture and the immunological characteristics, 58 candidiasis patients differing by the severity and dissemination of the disease were examined. Chronic candidiasis of the skin and mucous membranes, the most severe and disseminated form of the disease, is associated with a decrease in the number of T-lymphocytes and changes in their subpopulations, as well as high titers of Candida albicans antigen and antibodies to it in blood sera. The immune system of patients with visceral candidiasis and chronic vulvovaginal candidiasis was similar to that of healthy persons in the characteristics under study. Immediate and mixed hypersensitivity occurred in candidiasis patients more frequently than in healthy persons. In extremely severe forms of Candida infection immediate hypersensitivity prevailed.