[PARTICIPATION OF AROMATIC MICROBIAL METABOLITES IN THE DEVELOPMENT OF SEVERE INFECTION AND SEPSIS.]

Background Actuality of the problem of infection in critically ill patients remains high. Addition a local infectious processes have a tendency to rapid generalization and sepsis. Neutrophilsfunction failure plays a key role in the patho- genesis of infectious complications and sepsis. The hypothesis about the role of microbial metabolites in the formation of immunosuppression, in particular in the inhibition of phagocytic activity of neutrophils as the first line of defense against bacteria is acquiring more and more supporters. Among the microbial metabolites aromatic compounds, such as phenylcarbinol acid (PCA) are of particular interest. Their concentration in the blood reaches a maximum in case of generalized infection, and correlates with mortality in patients in critical conditions, and PCA biological activity confirmed by earlier researches in vitro. The purpose of the study was to reveal the relationship of the PCA with the severity of bacterial infections and to assess the effect of PCA on neutrophils phagocytic activity. MATERIALS AND METHODS: At the first stage the levels of PCA (GCh-FID method) in the serum ofpatients (n=57) with a documented bacterial infection of varying severity, confirmed APACHE II and SOFA scales were analyzed comparing with healthy donors (n=72). During the second stage the effect of clinically-relevant concentrations of PCA on neutrophils phagocytic activity ofperipheral blood was analyzed in vitro. RESULTS: The levels ofphenylacetic (PAA), phenyllactic (PLA), hydroxyphenylacetic (p-HPAA) and hydroxyphenyllactic (p-HPLA) acids in patients were significantly higher than in healthy donors, reaching the highest values in severe infection. High direct correlation of PCA levels with the indicators ofscales APACHE II and SOFA was detected. A significant decrease in the number ofphagocytic neutrophils under the influence PAA, p-HPAA and PLA was identified in vitro experiment on average 11%. The most significant influence p-HPAA, PLA and p-HPLA have made on the intensity of the neutrophils absorption capacity reducing this parameter by an average of 26%. Also noted that phenylpropionic acid (PPC) which is in blood of healthy donors, disappears in critical ill patients. CONCLUSION: Phenolcarbonic acids level in the blood serum shows the severity of bacterial inflammatory process. Ex- periment in vitro shows that the PCA in clinically-relevant concentrations is able to suppress the neutrophilsphagocytic activity.

[DETERMINATION OF A FATTY ACIDS CONTENT IN BILIARY LIPIDS AS A PROGNOSTIC FACTOR FOR THE COMPLICATIONS OCCURRENCE IN PATIENTS AFTER OPERATIONS FOR AN ACUTE CHOLECYSTITIS IN A HIGH OPERATIVE­ANESTHESIOLOGICAL RISK].

Investigation was conducted with objective to establish a prognostic significance of the fatty acids content in biliary lipids for the complications occurrence in patients, operated on for an acute cholecystitis, while presence of high operative­anesthesiological risk. A fatty acids content in biliary lipids was analyzed in 64 patients in presence of moderate (grade ІІ) and severe (grade ІІІ) course of the disease. Coefficients К1 and К2 for prognostication of postoperative purulent­inflammatory complications occurrence were elaborated, their threshold level (К1=0.59, К2=1.8) was established. There was established, that excess of the threshold meaning of К1 and К2 in a bile of patients, suffering an acute cholecystitis while presence of high operative­anesthesiological risk, constitutes a prognostic criterion for the postoperative purulent­inflammatory complications occurrence

[Effect of the nerve growth factor mimetic GK-2 on post-resuscitation expression of neurotrophic factors].

THE OBJECTIVE: to elucidate an influence of nerve growth factor mimetic GK-2 on the expression of neurotrophic factors and the process of neuronal death after ischemia-reperfusion. Materials and methods. Adult white male rats underwent cardiac arrest for 12 minutes, followed by resuscitation. 10 rats were injected GK-2 (Img/kg i/ρ) at 30 minutes and 48 hours after resuscitation. 10 untreated animals received equivalent doses of saline. The control group consisted of sham-operated animals (n = 10). On the 7th postoperative day the total density of hypoxia-sensitive cerebellar Purkinje cells was determined by morphometric analysis. Immunohistochemical study of proteins FGFb, NT4, BDNF was performed by indirect peroxidase-antiperoxidase method using primary polyclonal antibodies. The number of neurons with different expression levels of the neurotrophic factors was determined. RESULTS: In the post-resuscitation period the neuronal loss was detected in untreated animals. Namely NT4-negative, FGFb-negative and BDNF-negative cells died. GK-2 had no effect on the expression level of FGFb and NT4, however, promoted an increase in the expression level of BDNF. Initiating the expression of BDNF in neurons that were not previously producing this factor, GK-2 prevents the development of post-resuscitation neuronal death. Obtained facts lead to the conclusion that one of the mechanisms of neuroprotective action of nerve growth factor mimetic GK-2 is its ability to activate the expression of BDNF in nerve cells.

[INHALED ANTIBIOTICS IN TREATMENT OF NOSOCOMIAL PNEUMONIA].

Nosocomial pneumonia is the most common infection in intensive care units. Currently the problem of resistance of noso-comial pathogens to miost of antibiotics is crucial. Using of inhaled antibiotics in combination with intravenous drugs is eff ective and safe method for treatment of nosocomial pneumonia. The literature review describes current opportunities of ihhaled antibiotic therapy of nosocomial pneumonia, descriptions of drugs, the advantages and disadvantages of this treatment. Special attention is paid for using inhaled aminoglycosides for nosocomial pneumonia.

Effectiveness of GK-2, a Nerve Growth Factor Mimetic, in Preventing Post-Resuscitation Changes in the Brain.

Neurological status was assessed and the numbers of neurons per 1 mm in pyramidal neuronal layer length in CA1 and CA4 hippocampal fields and cerebellar Purkinje cells were evaluated in albino male rats on post-resuscitation day 14 after 12-min cardiac arrest. Intraperitoneal administration of GK-2 (1 mg/kg 30 min after resuscitation and within the next 3 days with an interval of 24 h) accelerated neurological recovery of the animals, sharply reduced the intensity of cerebellar Purkinje cell death, and prevented loss of pyramidal neurons in the hippocampus. These results show the effectiveness of systemic administration of the nerve growth factor mimetic GK-2 in improving structural and functional state of the brain in the post-resuscitation period. This opens new prospects of its use for prevention and correction of post-hypoxic encephalopathy.

Disorders in the Morphology and Nanostructure of Erythrocyte Membranes after Long-term Storage of Erythrocyte Suspension: Atomic Force Microscopy Study.

Disorders in the erythrocyte morphology and structure of their membranes during long-term storage of erythrocyte suspension (30 days at 4°C) were studied by atomic force microscopy. The morphology and nanostructure of erythrocyte membranes, biochemical parameters, ion exchange parameters, and hemoglobin spectra were recorded. The transformation of erythrocyte morphology and destruction of their membranes were observed throughout the storage period. Irreversible forms of spheroechinocytes and their fragments formed by the end of storage. The concentrations of potassium ions and lactate in solution of the blood preservatiive increased, while pH value decreased. Hemolysis detected by the erythrocyte "leakage" effect was observed starting from days 16-23 of storage.

Normal level of sepsis-associated phenylcarboxylic acids in human serum.

Previous studies showed that large amounts of phenylcarboxylic acids (PhCAs) are accumulated in a septic patient's blood due to increased endogenous and microbial phenylalanine and tyrosine biotransformation. Frequently, biochemical aromatic amino acid transformation into PhCAs is considered functionally insignificant for people without monogenetic hereditary diseases. The blood of healthy people contains the same PhCAs that are typical for septic patients as shown in this paper. The overall serum PhCAs level was 6 µM on average as measured by gas chromatography with flame ionization detection. This level is a stable biochemical parameter indicating the normal metabolism of aromatic amino acids. The concentrations of PhCAs in the metabolic profile of healthy people are distributed as follows: phenylacetic ≈ p-hydroxyphenyllactic > p-hydroxyphenylacetic > phenyllactic ≈ phenylpropionic > benzoic. We conclude that maintaining of stable PhCAs level in the serum is provided as the result of integration of human endogenous metabolic pathways and microbiota.

Clinical associations of host genetic variations in the genes of cytokines in critically ill patients.

Host genetic variations may influence a changing profile of biochemical markers and outcome in patients with trauma/injury. The objective of this study was to assess clinical associations of single nucleotide polymorphisms (SNPs) in the genes of cytokines in critically ill patients. A total of 430 patients were genotyped for SNPs in the genes of pro- (IL1B, IL6, IL8) and anti-inflammatory (IL4, IL10, IL13) cytokines. The main end-points were sepsis, mortality and adult respiratory distress syndrome (ARDS). We evaluated the dynamic levels of bilirubin, blood urea nitrogen, creatine kinase, creatinine and lactate dehydrogenase in five points of measurements (between 1 and 14 days after admission) and correlated them with SNPs. High-producing alleles of proinflammatory cytokines protected patients against sepsis (IL1B -511A and IL8 -251A) and mortality (IL1B -511A). High-producing alleles of anti-inflammatory cytokines IL4 -589T and IL13 431A (144Gln) were less frequent in ARDS patients. The carriers of IL6 -174C/C genotypes were prone to the increased levels of biochemical markers and acute kidney and liver insufficiency. Genotype-dependent differences in the levels of biochemical indicators gradually increased to a maximal value on the 14th day after admission. These findings suggest that genetic variability in pro- and anti-inflammatory cytokines may contribute to different clinical phenotypes in patients at high risk of critical illness.

[Influence of Microbial Metabolites of Phenolic Nature on the Activity of Mitochondrial Enzymes].

The aim of this work was to study the effect of microbial metabolites of phenolic nature on the activity of enzymes of the tricarboxylic acid cycle in isolated mitochondria, and determine metabolites of the tricarboxylic acid cycle as potential biomarkers of mitochondrial dysfunction in the blood of patients with sepsis. It is shown that microbial metabolites of phenolic nature have an inhibitory effect on the activity of dehydrogenases, determined by the reduction of dichlorophenolindophenol and nitroblue tetrazolium in liver mitochondria and liver homogenates. This effect is more pronounced in oxidation of the NAD-dependent substrates than succinate oxidation, and at lower concentrations of microbial metabolites than inhibition of respiration. By gas chromatography-mass spectrometry it was found that the content of the tricarboxylic acid cycle metabolites in the blood of patients with sepsis decreased compared to healthy donors. The data obtained show that the microbial phenolic acids can contribute significantly to the dysfunction of mitochondria and suppression of general metabolism, characteristic of these pathologies.

[Pathogenesis and target therapy of acute respiratory distress syndrome].

The paper summarizes results of experimental studies and clinical observations of the pathogenesis and effectiveness of respiratory, non-respiratory and pharmacological treatment methods for acute respiratory distress syndrome caused by direct and indirect damaging factors. The article deals with differences and peculiarities of morphological changes and lung functional disorders, clinical, laboratory and instrumental signs of various origins in ARDS and justifies necessity of differential diagnosis and differential treatment of ARDS, depending on the reasons for its development. Furthermore the article discusses an algorithm for differential diagnosis and differential treatment for ARDS caused by direct and indirect damaging factors.

[Cerebral and extracerebral insufficiency in patients with poor prognosis of surgical treatment of cerebrovascular circulation disorders of hemorrhagic type].

Objective of the study was to investigate cerebral and extracerebral insufficiency in survived and died patients after surgical treatment of hemorrhagic stroke for outcome prognosis and decision making on postoperative intensive care. The study included 224 patients after transcranial surgery for hemorrhagic stroke. 119 patients survived and 105 patients died SOFA scale and SIRS criteria were used to assess severity of the patients state. Hemorrhagic stroke before the operation was accompanied with cerebral insufficiency (Glasgow Coma Scale < or = 9) in 19% of patients. Lungs and kidneys dysfunction (SOFA = 1-2) were common. In the early postoperative period 54% of patients had cerebral insufficiency and 36%--organ dysfunction. Acute cerebral insufficiency was closely associated with systemic inflammatory response. Severity of organs (heart, lungs, kidneys) failure and SIRS correlated with GCS score in early postoperative period. There was close correlation between MODS and SIRS scores in survived patients and there wasn't such phenomenon in died patients. This indicates leading role of CNS in homeostasis regulation. SOFA scale using for express diagnosis of perioperative complications is useful for providing adequate intensive therapy.

[Development of neurocritical care].

In this review we provide the definition, goals and objectives of neurocritical care, evaluation of brief history of its development. Mechanical ventilation, intracranial hypertension, neuromonitoring as underlying basics of neurocritical care approaches are discussed. The main types of pathology and specific methods used in neurocritical care units are discussed. The results of our own research on brain death--the development of national criteria; for Guillain-Barre syndrome--a double decrease in the length of mechanical ventilation and in 2.5 times of the recovery time for independent walking ability; on diphteric polyneuropathy--reduced by 11 times mortality compared with nation-wide indicators of nontraumatic persistent vegetative state--the development of diagnostic and predictive neurophysiologic criteria are demonstrated. Research data of multiple organ disfunction syndrome in severe stroke are described. Further development of neurocritical care is being discussed.

[Pathophysiology of recruit ventilation and its effect on the respiration biomechanics (review of the literature)].

In patients with acute respiratory distress syndrome (ARDS) lungs consist of aeration zones and zones of alveolar collapse, which lead to intrapulmonary shunting and hypoxemia. ALV may increase alveolar collaps and potentially lead to lung damage arising out of displacing surfaces tension between aerated and collapsed lung parts and re-closing and opening of the alveoli. Based on the strategy of the "open lung" the recruitment maneuver can be used to achieve the alveolar recruitment, while PEEP prevents alveolar collapse. There are many recrutment types, including a long inflated to a higher pressure, reccuring breath, gradual increase of PEEP, and PIP etc. Data on the use of recruitment in ARDS patients, show varied results with increased efficiency in patients with early ARDS, good complaisance of the chest and extrapulmonary Genesis of lung damage. In this review, we discuss the pathophysiological basis of recruitment use, recent evidence and contradictions of the application of this method.

[Surgical treatment of duodenal fistulae].

The experience of surgical treatment of 10 patients with external intestinal fistulae was analyzed. The plot of the operation was to exlude the duodenum from the digestion by performing the retrocolic gastroenteroanastomosis after Roux or on the ultrashort loop. Early operation prevent septic complications and electrolyte disbalance. Duodenal fistulae, developed on the background of pancreonecrosis or after endoscopic retrograde cholangiopancreaticography, necessitate the external choledochial drainage.

Reversible zinc-induced injuries to erythrocyte membrane nanostructure.

Zinc-induced injuries to red blood cell membrane nanostructures at different zinc concentrations were studied by atomic force microscopy. In order to distinguish the intrinsic characteristics of membrane nanostructures, the membrane surfaces were represented by three orders using 3D Fourier transform. Increasing the concentrations of zinc ions modified the pattern of induced injuries: their depths and diameters and their number on the membrane surface test area increased. The injuries and their distribution for each order of membrane surface were analyzed. Albumin restored membrane nanosurface.

[Genetic study of predisposition to community-acquired pneumonia].

The study included 243 patients with acute community-acquired pneumonia and 173 healthy subjects. The following candidate loci were used to investigate genetic variability: 3 sites of CYP1A1, GSTM1, GSTT1, GSTP1, ACE gene of the rennin-angiotensin system, chemokine receptor gene CCR5. Enhanced predisposition to pneumonia was shown to be characteristic of homozygotes in deletion at the ACE locus (OR = 1.8; p = 0.013), carriers of normal alleles of the GSTM1 locus (OR = 1.7; p = 0.010), and homozygotes in allele 606T of the CYP1A1 gene (OR = 1.6; p = 0.020).

[Hemodynamic efficacy of modified gelatin in patients with acute blood loss and sepsis].

The paper presents the results of a clinical trial using the new modified gelatin-based drug hemofusin to correct dyshidria in patients with blood loss and sepsis. Based on the findings, the authors provide evidence for the high clinical efficacy of this drug and the expediency of its incorporation into a complex of infusion therapy in this patient contingent.

[The role of released membrane microparticles in pathogenesis of systemic inflammatory response].

The aim of this work was to evaluate contribution of released membrane particles (RMP) to the development of systemic inflammatory response (SIR) after aortocoronary bypass grafting (ACBG). The number of RMP carrying surface adhesion molecules, CD62L, CD62P, CD62E, was shown to increase in the early postoperative period in parallel with the enhancement of lymphocyte plasma membrane blebbing and elevation of cytokine levels in peripheral blood. It is concluded that (1) activation of plasma membrane blebbing in peripheral blood cells underlies the appearance of RMP in circulation; (2) increased number of RMP expressing CD62L, CD62P, CD62E is a marker of intercellular communication associated with the development of SIR and suggests new mechanisms of RMP involvement in the reaction of organism to massive surgical injury.

[Lung morphological changes in premature neonates with hyaline membrane disease due to the use of exogenous surfactants during mechanical ventilation].

A complex of studies was used in 4 groups of premature babies to study lung tissue morphological changes in hyaline membrane disease, by applying exogenous surfactants during mechanical ventilation. Background diseases, pre- and intranatal risk factors, the babies' longevity, and the specific features of lung tissue and forming hyaline membranes were ascertained. Exogenous surfactants were found to have a blocking effect on the formation of hyaline membranes under mechanical ventilation.