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Landscape of transition of microorganisms to internal organs during the experimental bacterial translocation
Table 2. - Comparative characteristics of patients main and control group
Parameters Main group (n=38) Control group (n=31)
Sex male 25 (65,8%) 17 (54,8%)
female 13 (34,2%) 14 (45,2%)
Age, years 48,9 [42,5-58,7] 48,0 [38,2-53,7]
RI, mmHg 143,0 [127,4-173,2] 137,0±18,2
LIS, points 2,6 [2,1-2,91] 2,7 [2,45-3,1]
Dynamics of changes in RI are presented in Figure 1.
Fig. 1. Dynamics
As shown in the graph presented, in patients of the main group respiratory index increased much faster than the control (p <0,05). This suggests that early and mandatory implementation of 'Recruitment' maneuver in patients with ARDS on the background of obesity is already possible to eliminate atelectasis during the 1st day of mechanical ventilation has led to the progressive improvement of oxygenation. Number of days on mechanical ventilation, decreased from 11,1 ± 10,2 to 8,2 ± 3,6 days. The mortality rate decreased from 42% (corresponding to world statistics) to 15.8%.
of respiratory index Conclusions
Obesity — a risk factor for ARDS and severe course in patients with community-acquired pneumonia. Early transfer of patients with bilateral infiltrates on chest radiograph and obesity in the ICU, early non-invasive ventilation and early transfer to mechanical ventilation (translation criteria: PaO2/FiO2 <175 mmHg after 1 hour conducting NIV) c mandatory implementation of maneuver 'Recruitment' can effectively prosthesis lung function in these patients, helping to reduce the duration of mechanical ventilation and decrease mortality.
References:
1. Полушин, Ю. С. Вирусная пневмония грипп А (H1N1), осложненная ОРДС/Ю. С. Полушин [и др.]//Общая реаниматология. -2010. - № 3. - С. 15-22.
2. Чучалин, А. Г. Грипп: уроки пандемии//Пульмонология. Приложение: «Грипп A/H1N1: уроки пандемии». - 2010. - С. 3-8.
3. Incidence and outcomes of acute lung injury/G. D. Rubenfeld [et al.]//N Engl J Med. - 2005. - Vol. 353. - P. 1685-1693.
4. Mortality Rates for Patients With Acute Lung Injury/ARDS Have Decreased Over Time/M. Zambon, J.-L. Vincent//Chest. - 2008. -Vol. 133. - P. 1120-1127.
5. Светлицкая, О. И. Острое повреждение легких у пациентов с вирусно-бактериальной пневмонией на фоне избыточной массы тела и ожирения/О. И. Светлицкая, И. И. Канус//Медицинские новости. - 2013. - № 3. - С. 6-10.
6. Body mass index is associated with the development of acute respiratory distress syndrome/M. N. Gong [et al.]//Thorax. - 2010. -Vol. 65. - P. 44-50.
Suwonov Qayim Jahonovich, PhD, Senior scientific employee-researcher, Research Institute of Sanitation, Hygiene and Occupational Diseases Ministry of Health of Uzbekistan,
Landscape of transition of microorganisms to internal organs during the experimantal bacterial translocation
Abstract: The purpose of the scientific work was study of the landscape of microorga-nisms grown (LMG) from the internal organs of laboratory animals in experimental acute obstru-ction of the small (EAOSI) and large intestine (EAOLI). It has been established that Gram-ne-gative sticks, Gram-positive cocci and anaerobes were identified in the experimental model EAOSI. Anaerobic translocation low level of ability. With the increase in the duration of mic-roorganisms translocation (MT) the shutter all the members are equally spread. EAOSI there is a big difference between the recovered Staphylococcus spp and Enterococcus spp recognized as one of the main microbiological criteria.
Keywords: experimental obstruction, the small intestine, the large intestine, landscape of microorganisms, bacterial translocation, Gram-positive cocci.
As you know, representatives of the normal microflora of the human large intestine — in-digen and facultative organisms with balance disorders (dysbiosis), with an increase of permea-bility through the mucous membrane of the intestine, would be increased passage of viable mic-roorganisms in a variety of internal organs through the blood and lymph. "Bacterial translo-cation" (BT) this state, along with the name of microorganisms passing on these organs is called MT [2; 9; 10; 11].
There are different opinions about the importance of this phenomenon: the first suppor-ters consider it one of the pathogenetic links ofvarious diseases [1; 3; 7], while others will inter-pret this as a factor in protecting the organism [4; 6]. Taking into account the usefulness of the experimental studies, we started it.
Purpose of the work. The study and evaluation of the LMG from the internal organs of laboratory animals in (EAOSI) and (EAOLI).
Materials and methods. During implementation of our scientific research work, we strictly followed the biosecurity rules and principles of ethics in work with experimental material. A total of 368 surveys, white, wild mice, weighing 25 grams and higher, the age of 2-3 months. Feeding and care of laboratory animals, division into groups and preparation those for the experiments were carried out in the traditional methods.
We used an experimental model EAOS and EAOLI with the introduction of our modifi-cations proposed by Kruglyanskiy Y. M. [5] in the formation of an EAOS and EAOLI. Conducted 3 series of research work.
All laboratory animals divided into 4 groups: Group 1 — EAOSI, n=72; Group 2 — EAOC, n=72; Group 3 — animals, which were opened in the abdominal cavity, but not performed obtu-ration (group comparisons, n=72); Group 4 — intact laboratory animals (control group, n= 24). In turn, 1, 2 groups were divided into subgroups: 1a, 2a — EAOSI EAOC and lasted 24 hours (n=8); 1b, 2b — EAOSI and EAOC, lasted for 48 hours (n=8); 1c, 2c — EAOSI and EAOK, lasted for 72 hours (n=8).
These deadlines are selected as a result of the wall of the intestine obstruction mainly according to the pathological, morphological, clinical changes [3; 5].
Advanced bacteriological study was carried out with respect to microorganisms grown from the internal organs of laboratory animals. Identification and differentiation of the seeded microorganisms carried out by Bergey's [8]. To do this, the nutritional resources of the company «HiMedia» (India) were used.
Statistical analysis of results of biomedical research were carried out on the personal computers with processor "Pentium 4", using "Excel" special program.
Results and discussion. According to the deadlines of the experiment, LMG from the mesenteric lymph nodes (MLN) in the experimental model EAOSI were different. Among all st-rains, Escherichia spp were most common. Particularly attracted the attention that the 24 and 48 hours of experience was grown only Escherichia spp (n=10 and n=7, respectively) as a mono-culture.After formation of EAOSI, grown Gram-negative (Enterobacter spp, Citrobacter spp, Proteus spp), Gram-positive (Staphylococcus spp, Enterococcus spp) and anaerobes (Bac-teroides spp) as a monoculture not have been grown from the MLN of used laboratory animals.
The association of microorganisms has got a different situation. Although the experience of leadership in quantitative terms even if it is still owned by Escherichia spp (3<0.001) was observed in other microorganisms rawled.
If after 24 hours in strains of microorganisms in the form of the association of sprouts 45.6 ± 7.3% (n=21), Escherichia spp, here in after referred to as Proteus spp (17.4 ± 5.6%, n=8), Enterobacter spp (15.2 ± 5.3%, n=7), Staphylococcus spp (10.9 ± 4.6%, n=5), Citrobacter spp (8.7 ± 4.2%, n=4), Enterococcus spp (2.2 ± 2.1%, n=1). Bacteroides spp as monocultures and associations have not been grown.
After obturation, 48 hours later, growth of microorganisms increased in amount, but it remained similar to the above tendency. According to the the division of interest was as follows: Escherichia spp 47.4 ± 4.6% (n=55), Proteus spp 26.7 ± 4.1% (n=31), Enterobacter spp 8.6 ± 2.6% (n=10), Staphylococcus spp 7.8 ± 2.5% (n=9), Citrobacter spp 6.7 ± 1.8% (n=9), Enteroco-ccus spp 0.9 ± 0.8% (n=1).
Experience the next term (72 hours) quantitative microorganisms along with the more detached from their ratio to one another also changed. Proteus spp, Escherichia spp on the redu-ced rate of other microorganisms on a significant scale has not changed. 72 hours, depending on the level of MT in the following sequence: Escherichia spp (36.6 ± 4.2%, n=48), Proteus spp (21.4 ± 3.6%, n=28), Enterobacter spp (16.0 ± 3.2%, n=21), Staphylococcus spp (12.2 ± 2.9%, n=16), Citrobacter spp (11.8 ± 2.7%, n=16), Enterococcus spp (0.8 ± 0.7%, n=1), Bacteroides spp (0,9 ± 0,6%, n=1). It is noteworthy that, irrespective of the length of experience did not change the amount of Enterococcus spp, and Bacteroides spp. We believe that this EAOSI MT rate is low, it means that translocations ability and the ability of other Gram-sticks low against Gram-positive cocci. The evaluation of the intensity of BT EAOSI pathogenetic mechanism is based on the formation of an experimental model of EAOSI that this situation should be taken into account.
Experimental model EAOSI MLN has grown compared to the corresponding period of microorganisms (Figure 1) percent of strains of Escherichia spp other increases decreased with increasing duration of the experience. We believe that this form of association due to other microorganisms.
Experimental model EAOSI obturation the next 24, 48, 72 hours microorganisms rawled MLN from a lot of different changes in the liver retained. Liver only form of monoculture, Escherichia spp collected: 9 strain after 24 hours, 48 hours, 12 stamps, and 72 hours after 9 stamps. Other microorganisms only in the form of associations failed. Escherichia spp, 24 hours later, all the grown strains of 50.0 ± 10.2% (n=12), 48 hours after it was 43.1 ± 6.5% (n=25), 72 hours after the 47.5 ± 5.0% (n=47), respectively. Common interest between the difference is not statis-tically reliable (p>0.05).
Thus, the level of Escherichia spp met in the increase of the amount, depending on the duration of the experiment thought about it, measured in relation to the other microorganisms.
After 24 hours, Proteus spp and Staphylococcus spp respectively 16.7 ± 7.6% (n=4) and 20.8 ± 8.3% (n=5), established, 48, and 72 hours after they are respectively were as follows: 27.6 ± 5.9% (n=16), 12.1 ± 4.3% (n=7) and 20.2 ± 4.0% (n=20), 11.1 ± 3.2% (n=11). Enterobacteria (Enterobacter spp, Citrobacter spp), along with the number of microorganisms as defined above, interest rates can not compete. Enterococcus spp, and Bacteroides spp MLN parameters were similar results, that is, detecting significant changes in the risks associated with a period of EAOSI and reliable.
The results also differ in terms of the number of microorganisms on the divorce, or a ratio of one significant difference was detected. This member received compensation in the form of monoculture, mainly Escherichia spp. Identification of these microorganisms in the
Landscape of transition of microorganisms to internal organs during the experimantal bacterial translocation
form of the association are the advantages clearly visible. 48 hours after the association's stamp form of 20 and 9 (45.0 ± 11.1%), which is owned by Escherichia spp, 5 (25.0 ± 9.7%), Staphylococcus spp, and 4 (20.0 ± 8.9%), Proteus spp, and 2 (10.0 ± 6.7%), Enterobacter
spp. The rest of the microor-ganism (Citrobacter spp, Enterococcus spp, Bacteroides spp) identified this term. This trend re-mained after 72 hours. Treeless Citrobacter spp (9.1 ± 6.1%, n = 2), Enterococcus spp (4.5 ± 4.4%) were identified.
Figure 1. Experimental EAOSI MLN of the results of the study, compared to the corres-ponding period of sprouts and microorganisms,%
Lungs seedling also done a lot of Escherichia spp charged with distinction. Due to a lack of statistical analysis of the number of strains collected, we find it necessary to comment on all the numbers, but the trend of the previous commented exchange want to admit the rest.
Research at the next stage of our game was similar to the above research. Experimental model MLN strains of microorganisms from EAOSI to germinate, such as Escherichia spp. 24 hours later after obturation monoculture 6 stamp detached, the association is in the form of 23 stamps. This form of association for all strains of microorganisms to 47.9 ± 7.2%, respectively. Followed by Enterococcus spp (14.6 ± 7.2%, n=7), Proteus spp (12.5 ± 4.8%, n=6), Enterobacter spp (12.5 ± 4.8%, n=6), Citrobacter spp (8.3 ± 4.0%, n=4), Staphylococcus spp (4.2 ± 2.9%, n=2) busy. This term has not been identified Bacteroides spp. The main difference from the experimental EAOSI Enterococcus spp's a lot to be identified.
48 hours later a changed ratio of microorganisms. This term Escherichia spp (45.6 ± 5.6%, n=36), followed by Proteus spp (15.2 ± 4.0%, n=12), Enterobacter spp (12.7 ± 3.8% n=10) respectively. Other strains of Enterococcus spp slightly less (11.4 ± 3.6%, n=9) were collected. It is run pilot EAOSI less than the total number of strains, compared with 1.5 times (116 stamps 79 stamps) have been identified.
72 hours after the results are a little different, if EAOSI at the same time from the as-sociation, a total of 131 in the form of stamp collection, at 1.2 times this figure (160 strain). Es-che-richia spp, the total number of strains from 38.1 ± 3.8% (n=61), respectively. Lack of reliable compared to the previous terms of this indicator (P <0.05) after 72 hours of Enterococcus spp (17.5 ± 3.0%, n=28).
Each model offers 72 hours after the results of the comparative study of the landscape of the MLN has grown microbes in Figure 2.
Figure 2. Experimental EAOSI and 72 hours later after the obturation in the EAOSI, MLN of the microbes collected from the landscape, %
Figure 2 shows that both views of the model identified by microorganisms in the form of association clearly shows differences in the level of invention. In each case, the intensity of BT leading difficult to distinguish microorganisms.
Results of experimental liver has LMG MLN figures are similar. At the same time, micromonocultural concerning the association found that only 24 and 48 hours respectively 3 (Escherichia spp) and 5 (Escherichia spp) case.As the leadership of the association, in the form of MLN Escherichia spp. Escherichia spp 24 hours after 50.0 ± 11.2% (n = 10) identified in the case, other microorganisms (Proteus spp, and Enterobacter spp, Citrobacter spp, Enterococcus spp), 50.0 ± 11.2%. Unlike EAOSI not identify Staphylococcus spp.
48 hours after the results were completely different, the leadership is still Escherichia spp (46.7 ± 6.4%), followed by Enterococcus spp 25.0 ± 5.6%, n=15) and Proteus spp (15.0 ± 4.6%, n=9) out. These results suggest that changes in the parameters of the EAOSI trend similar to that of the case (58 stamps of 60 stamps), were differences in the ratio of one of the microorganisms.
If EAOSI Proteus spp 27.6 ± 5.9% (n=15), Staphylococcus spp 12.1 ± 4.3% (n=7), Enterococcus spp 3.4 ± 2.4% (n=2) is charged, this is one of the parameters of a credible difference (p<0.05) respectively 15.0 ± 4.6% (n=9), 3.3 ± 2.3% (n=2) and 25.0 ± 5.6% (n=15) (Figure 3).
Figure 3. Experimental EAOSI and after
Experience the next term (72 hours), the number has grown strains compared to the previous period to 2.0 times (119 stamps 60 stamps).The percentage of microorganisms different from each other, positive results have been obtained. At the same time, Esch-erichia spp liver microorganisms him after 48.7 ± 4.6%, respectively. This option MLN has grown Escherichia spp, which is more than 1.3 times the amount of reliable, EAOSI index (p>0.05).
At the same time Enterobacter spp (6.7 ± 2.3%, n = 8), Citrobacter spp (5.9 ± 2.2%, n=7), Proteus spp (16.8 ± 3.4%, n=20), Bacteroides spp (1.7 ± 1.2%, n=2) compared to the same pe-riod of the indicators EAOSI whether significant difference (p>0.05), Staphylococcus spp (1.7 ± 1.2% and 11.1 ± 3.2%) and Enterococcus spp (18.5 ± 3.8% and 2.0 ± 1.4%) on the parameters of the difference was significant (p<0.001).
Identification of a spleen is also dominated by the amount of microorganisms Escherichia spp, and 48 hours later in the form of the Association charged him 7 of 14 stamps. Enterobacter spp, Citrobacter spp, Staphylococcus spp not identified, but even if it is less than 72 hours after their rawled. 48 and 72 hours in other strains of Proteus spp and Enterococcus spp significantly greater in the head. Thus, the increase in the number of Enterococcus spp can be seen as a remar-kable situation.
MLN results obtained from the lungs, liver and spleen variables are no similar laws simi-lar trend Escherichia spp other microorgan-
the obturation 48% of the liver has LMG
isms to germinate.Due to a lack of statistical analysis of the number of strains collected from all found that the numbers are interpreted in an illegal.
Conclusions. 1. It has been established that Gram-negative sticks, Gram-positive cocci and anaerobes were identified in the experimental model EAOSI. Experience all the time (24, 48, 72 hours), the amount of Escherichia spp other strains % part. Across all strains % enterobacteria. The ability to translocation against Gram-negative bacteria are Gram-positive cocci 3-4 times higher.
2. Anaerobic (Bacteroides spp) tranlocation low level of ability and experience with in the quantitative aspects of change.
3. All members of the growing MT the shutter time in a uniform distribution, and all indicators MLN clearly seen in the liver, spleen, and in particular the lungs, the microorganisms in the form of monoculture and association met thought about it, was not evident in the intensity of BT.
4. Experimental model of EAOSI LMG MLN been identified by credible distinction, which is, first of all characterized by an increase in the number of strains allocated 2.0 times, second 48, and 72 hours later, Enterococcus spp, Escherichia spp put to the next problem.
5. Experimental EAOSI there is a big difference between the recovered Staphylococcus spp and Enterococcus spp recognized as one of the main microbiological criteria.
References:
1. Almagambetov K. K., Bondarenko V. M. Simulation of translocation of intestinal microflora on conventional animals//Medical journal - Moscow, 1991; 8: 11-7. (In Russian).
2. Galeev Y.M, Popov M. V., Salato O. V. Methods of study of the propagation of bacterial cells//Siberian Journal of Medicine. - Irkutsk, 2011; 3: 18-23. (In Russian).
3. Gostishchev A. N. Afanasiev Y. M. Kruglyanskiy D. N., Sotnikov V. K. Bacterial translocation in acute intestinal obstruction//Journal ofMedical Sciences - Moscow, 2006; 9-10: 34-8. (In Russian).
Using immunocorrection therapy in patients with chronic pancreatitis
4. Gritsenko VA Properties of E. coli isolated from mice with bacterial translocation after im-mobilization stress//Medical journal -Moscow), 2000; 2: 37-41. (In Russian).
5. Kruglyanskiy Y. M. Bacterial translocation with obstructive ileus (experimental research): Abstract. Dis. cand. med. sciences. - Moscow, 2007; 24 p.
6. Nikitenko V. I., Tkachenko E. I., Stadnikov A. A. Translocation of bacteria from the gast-rointestinal tract -a natural defense mecha-nism//Experimental and clinical gastroenterology - Moscow, 2004; 1: 48-53. (In Russian).
7. Nurmukhamedov H. K. Bacterial translocation in infectious-toxic shock in infants//Infection, immunity and pharmacology - Tashkent, 2004; 1: 103-4. (In Russian).
8. The determinant of bacteria Burgi. Edited by J. Holt, N. Krieg, Sneath P., J. Staley, Williams C. - Moscow: "Mir." 1997; Vol. 1-2.
9. Ergashev V. A., Nuraliev N. A. The phenomenon ofbacterial translocation and location of microorganisms in its formation//Infection, immunity and pharmacology - Tashkent, 2014; 3 (2): 236-9. (In Russian).
10. Berg R. D., Garlington A. W. Translocaision of certain indigenous bacteria form the gastroin-testinal tract to the mesenteric lymph nodes and other organs in a gnotobiotic mouse model//Infection and immunology, 1979; 23 (2): 403-11.
11. Berg R. D. Bacterial translocation from the intestines//Jikken Dobutsu, 1985; 34 (1): 1-16.
Suleymanov Suleyman Fayzullaevich, PhD, Senior Researcher, Head of the Chair of Microbiology and Phthisiology of the Bukhara State Medical Institute, Bukhara, Uzbekistan
E-mail: [email protected]
Using immunocorrection therapy in patients with chronic pancreatitis
Abstract: The immune system studied in 36 patients with chronic pancreatitis (CP) and 32 healthy individuals. In patients with CP showed a deficiency of T-lymphocytes and subset tension humoral immunity and cytokine profile. Using Thymopti-num (dose of 0.8-1.0 mg per course) in conjunction with conventional treatment in patients with CP led to an increase in cellular immunity and stabilization of cytokine levels.
Keywords: the immune system, T- and B-link immunity, cellular immunity, humoral immunity, link, immunotherapy, pancreatitis.
Changes in the environment, technology, food industry, lifestyle and spreading "western food" are the reason of growing the diseases of the pancreas. Over the past 30 years, marked by the global trend to an increasing in the incidence of acute and chronic pancreatitis (CP) is more than 2 times [2; 3; 10].
CP prevalence, increasing morbidity and temporary disability due to disability is an important social and economic problem of modern medicine. In the structure of morbidity gastrointestinal organs CP is from 5.1 to 9%, and in general clinical practice — from 0.2 to 0.6% [3, 5-7; 12].
Inflammation in the pancreas can develop due to malfunction of the immune system, which are based on allergic reactions, as well as response to bacterial factor [1; 5; 8].
However, immune disorders and their correction in patients with CP are still poorly studied to the present time.
The purpose of the work — studying of the immune system parameters and conduct immunocorrecting treatment in patients with CP.
Materials and methods. 36 patients were examined (3365 ages) with a diagnosis of CP. The diagnosis was carried out on the basis of complaints, medical history, and objective laboratory tests, instrumental data: ultrasound, fibrogastroduodenoscopy, survey radiography of abdominal organs. The control group consisted of donors from 32 healthy subjects (25-55 ages).
The concentration of serum immunoglobulins (SI) classes A, M and G were determined by radial immunodiffusion (Mancini G., 1965).
The parameters of cellular immunity (T-lymphocytes and a subpopulation, B-lympho-cytes) were identified using monoclonal antibodies (LLC "Sorbent Service", Russia) [4]. Quanti-fication
of levels TNF-a, IL-6, IL-4 in serum performed using reagents set ProCon (LLC "Protein contour", St. Petersburg) by ELISA.
Immunotherapy was carried out in 15 patients. Thymoptinum (Uzbekistan) was used as an Immunological drug. 0.8-1.0 mg per treatment (dose 100 mg/day for 8-10 days). The indicators ofimmu-nity was studied twice: before — and after 1 month after treatment).
Results and discussion. In patients with CP found immunodeficiency cell component: 0.7-times whatever suppression of the total lymphocyte pool — T (CD3) — 35.3 ± 2.6% as compared with the control group — 52.4 ± 1.8% (p<0.001); 0.8-fold decrease in the absolute number of T (CD3)-cells (p<0.05).
Also determined the oppression subpopulations of T-lymphocytes, have the helper-suppressor function — Th (CD4) — 29.5 ± 1.1% (p <0.001) and 341.8 ± 32.1 cells/1 mcl blood (p<0.001) (control in 36.5% ± 0.7 and 616.4 ± 44.3 cells/1 mcl of blood, respectively), the contents of Ts (CD8) — 13.8 ± 1.4% (p<0.05) and 127.3 ± 9.8 cells/1 mcl blood (p<0.01)
On the side of B (CD19) — cell link, opposite, the tendency to increase as the relative parameter — 20.6 ± 2.3% (p<0.05), which was 1.4 times higher than those ofthe control group values, such and totally — 1.7-fold increasing — 385.8 ± 33.4 cells/1 mcl of blood (in the control — 230.1 ± 26.7 cells/1 mcl of blood).
An appreciable activation of B-cell immunity against the background suppression of T cells in CP reflected in the SI spectrum. For example, attention is drawn to the increasing in IgA production to 3.97 ± 0.41 g/l (p<0.05), which may be a reflection of the organism immune adjustment CP patients in response to enzymatic intoxication.
It was found authentically a high content of IgG — 22.42 ± 0.75 g/l (p<0.001) (in control of 15.9 ± 0.94 g/l). IgM concentra-
