Dr.Biol., Associate Professor O.N. Oparina¹
Dr. Hab., Associate Professor A.A. Pashin¹
PhD, Associate Professor J.V. Toma^1
1Penza State University, Penza

Keywords: endotoxin, endotoxin aggression, etiological factors.

Background. Endotoxin is known to activate the hemostatic system [2, 3] in certain physiological conditions due to its Hageman factor activation capacity, and interact with receptors of granulocytes and platelets [4]. One of the endotoxinemia rating methods in case of a gram-negative sepsis is based on endotoxin rating in the blood circulation system with antibacterial immunity tests in athletic trainings that provide the means to rate the training system efficiency and detect latent diseases in need of special diagnostics and pathogenetic therapy.

Objective of the study was to analyze the etiological factors of influence on a sport-specific endotoxin aggression.

Methods and structure of the study. We tested endotoxin of gram-negative intestinal bacteria in the blood plasma, plus antibacterial humoral immunity in the 18-20 year-old athletes (n=79) versus the Reference Group of virtually healthy 20-40 year-olds (RG, n=30). We used the systemic endotoxinemia and antiendotoxin immunity (AEI) test method developed by the Institute of General and Clinical Pathology (RF Patents No. 2011993, 2088936, 2093825); and rated the antibacterial humoral immunity by an enzyme immunoassay to find contents of the antibacterial antibodies to the following antigens of opportunistic bacteria: Pseudomonas aeruginosa, Escherichia coli, Proteus, bacteroids, Klebsiella, bifidumbacteria and Candida antigens. The test data were processed by the MS Excel 5.0 statistical toolkit, with the data array differences rated significant at p<0.05.

Results and discussion. The relevant reference literature reports correlations of endotoxinemia with thrombocytopenia [1, 4]; and we matched in this context the peripheral blood platelets in EG-1 individuals tested with blood plasma endotoxin of 0.3-1.25 EU/ ml (close to the physiological norm of 0-1.0 EU/ ml) versus that in EG-2 tested with blood plasma endotoxin of 1.5-5.0 EU/ ml: see Table 1 hereunder. Note that EG-1 was tested with increased platelets indicative of "irritation" i.e. endotoxin stimulation of the myelocytic bone marrow lineage [1].

Table 1. Peripheral blood platelets versus endotoxin in athletic EG-1 and EG-2

Note: *р≤0.05

EG-2 was tested with the platelet levels 25% and 30% lower than the standard and the EG-2 test rates, respectively, close to the bottom of the standard range which has been established by the decades-long clinical tests of virtually healthy samples. The clinical data shows that when endotoxin averages 2.53±0.13 EU/ ml, it may be indicative of the myelocytic bone marrow lineage reserves being close to depletion. Such imbalance may result in an adaptation failure (de-adaptation) under pressure, with the granulocyte link response being almost the same as response from platelets.

It should be noted that 42 athletes (53.2% of the EG) were tested with endotoxin aggression without expressed clinical manifestations, since none in the EG was tested with body temperature above 36.7⁰С. This may mean that the subjects may be diagnosed with a chronic endotoxin aggression in the “endotoxin tolerance” stage. Presently “endotoxin tolerance” may be defined as the purely experimental notion for the situations when no temperature growth is found in response to endotoxin in the blood.

Of special interest for us in the above context was an attempt to find which of the gram-negative intestinal bacteria contribute to the endotoxin aggression progress in the group. We used for this purpose an indirect serological test to rate antibodies to microbial sources of endotoxin aggression - most probable in our opinion. In addition, we rated antibodies to Candida and bifidobacteria, to detect secondary immunodeficiency (in the first case) and potential damage to the intestinal barrier permeability (second case), since bifidobacteria are known to be highly adhesive to the epithelium of the colon mucosa and compete there with gram-negative bacteria.

Based on the above test data, we classified the EG into EG-1a and EG-2b with the normal and high antibodies to bifidobacteria in the blood plasma. Thus EG-1a was tested with 3.6 to 4.6 μg/ ml (4.10 ± 0.50 on average); and EG-2 with 6.1 to 18.2 μg/ ml (8.90 ± 0.70 on average) of antibodies to bifidobacteria. Of special interest is the fact that EG-1a and EG-2a differed also in the blood plasma endotoxin, titers of antiendotoxin antibodies and antibacterial antibodies.

Every group with normal antibodies to bifidobacteria rates was tested with increased blood plasma endotoxin and low titers of antiendotoxin antibodies. These groups were also tested with significant growth of antibodies to Candida antigens (save for the weightlifting group) – that may be indicative of candidal dysbiosis as a marker of secondary immunodeficiency. Subjects with the high antibodies to bifidobacteria were tested with significantly (p <0.05) high blood plasma endotoxin and high titers of antibodies to E. coli 014; plus significantly (p <0.05) lower titers of antibodies to Re-glycolipid.

The above changes are assumedly typical for increased translocation of bacteria and/ or endotoxin from the intestine into the bloodstream due to intestinal dysbiosis associated with a deficiency of bifidobacteria. Individuals with high antibodies to bifidobacteria were tested with significant changes in antibodies to antigens of gram-negative bacteria: Pseudomonas aeruginosa (triathletes and basketball players), Proteus (basketball players), bacteroids (weightlifters and basketball players), Klebsiella (weightlifters, triathletes, track and field athletes and basketball players), and Candida (weightlifters, triathletes and basketball players) – versus those tested with normal antibodies to bifidobacteria. The detected variations in the antibacterial antibodies may be due to dysbiosis triggered by sport-specific stressors and contributing to the endotoxin aggression progress.

Conclusion. The study found a direct correlation between the blood plasma endotoxin and antibodies to bifidobacteria indicative of a high permeability of the intestinal barrier. The group with high antibodies to bifidobacteria was diagnosed with endotoxin aggression of different sport-specific etiologies that exposes the group to a high risk of an adaptation failure (de-adaptation).

References

1. Oparina O.N. Biological properties of intestinal microflora endotoxin. Modern scientific research and innovation. January 2014. No. 1 [Electronic resource]. Available at: http://web.snauka.ru/issues/2014/01/31034
2. Permyakov N.K., Yakovlev M.Yu. Pathology of digestive system and systemic endotoxinemia. Arkhiv patologii. 1989. v. 51. No. 12. pp. 74-79.
3. Permyakov N.K., Anikhovskaya I.A., Anokhin V.A. et al. Diagnostics of systemic endotoxinemia. Actual problems of general and specific pathology. Proceedings of Research Institute of Human Morphology of the Russian Academy of Medical Sciences. Moscow, 1993. pp. 7-10.
4. Yakovlev M.Yu. Systemic endotoxinemia in human physiology and pathology. Doct. Diss. Abstr.. Moscow, 1993. 55 p.

Corresponding author: ffkpqpu@mail.ru

Abstract

Objective of the study was to analyze the etiological factors of influence on a sport-specific endotoxin aggression.

Methods and structure of the study. We tested endotoxin of gram-negative intestinal bacteria in the blood plasma, plus antibacterial humoral immunity in the 18-20 year-old athletes (n=79) versus the Reference Group of virtually healthy 20-40 year-olds (RG, n=30). We used the systemic endotoxinemia and antiendotoxin immunity test method developed by the Institute of General and Clinical Pathology (RF Patents No. 2011993, 2088936, 2093825); and rated the antibacterial humoral immunity by an enzyme immunoassay to find contents of the antibacterial antibodies to the following antigens of opportunistic bacteria: Pseudomonas aeruginosa, Escherichia coli, Proteus, bacteroids, Klebsiella, bifidumbacteria and Candida antigens. The test data were processed by the MS Excel 5.0 statistical toolkit, with the data array differences rated significant at p<0.05.

Results and conclusions. The study found a direct correlation between the blood plasma endotoxin and antibodies to bifidobacteria indicative of a high permeability of the intestinal barrier. The group with high antibodies to bifidobacteria was diagnosed with endotoxin aggression of different sport-specific etiologies that exposes the group to a high risk of an adaptation failure (de-adaptation).