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C. (P.)
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Ðåäàêö³éíà ðàäà (Editorial Board)
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1
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Çì³ñò (Contents)
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2-9
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ÎÃËßÄÈ (REVIEWS)
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MOLECULAR DYNAMICS STUDY OF INTERACTIONS OF POLYMYXIN B3 AND ITS ALA-MUTANTS WITH LIPOPOLYSACCHARIDE
Lisnyak Yu. V.
Introduction.
Emergence of nosocomial bacterial pathogens (especially Gram-negative
bacteria) with multiple resistance against almost all available
antibiotics is a growing medical problem. No novel drugs targeting
multidrug-resistant Gram-negative bacteria have been developed in
recent years. In this context, there has been greatly renewed interest
to cyclic lipodecapeptides polymyxins. Polymyxins exhibit rapid
bactericidal activity, they are specific and highly potent against
Gram-negative bacteria, but have potential nephrotoxic side effects. So
polymyxins are attractive lead compounds to develop analogues with
improved microbiological, pharmacological and toxicological properties.
A detailed knowledge of the molecular mechanisms of polymyxin
interactions with its cell targets is a prerequisite for the purposeful
improvement of its therapeutic properties. The primary cell target of a
polymyxin is a lipopolysaccharide (LPS) in the outer membrane of
Gram-negative bacteria. The binding site of polymyxin on LPS has been
supposed to be Kdo2-lipid A fragment. Methods.
For all molecular modeling and molecular dynamics simulation
experiments the YASARA suite of programs was used. Complex of
antimicrobial peptide polymyxin Â3 (PmB3) with Kdo2-lipid A portion
of E. coli lipopolysaccharide was constructed by rigid docking
with flexible side chains of the peptide. By alanine scanning of
polymyxin Â3 bound to LPS followed by simulated annealing minimization
of the complexes in explicit water environment, the molecular aspects
of PmB3-LPS binding have been studied by 20 ns molecular dynamics
simulations at 298 K and pH 7.0. The AMBER03 force field was used with
a 1.05 nm force cutoff. To treat long range electrostatic interactions
the Particle Mesh Ewald algorithm was used. Results.
Ala-mutations of polymyxin’s residues Dab1, Dab3, Dab5, Dab8 and Dab9
in the PmB3-LPS complex caused sustained structural changes resulting
in the notable loss in stability of LPS complexes with Ala-mutants of
PmB3. The mutations disturbed the characteristic hydrogen-bond network
of PmB3-LPS complex. Ala-mutations of Dab1, Dab8 and Dab9 amino acid
residues of PmB3 destabilized PmB3-LPS complex to a greater extent: the
values of binding energy for these mutants showed increase and
large-amplitude irregular fluctuations. Conclusions.
Hydrogen bonding of polymyxin B with the lipopolysaccharide is an
important factor of the stability of PmB3-LPS complex. Detailed
knowledge of the peculiarities of molecular interactions of polymyxins
with its primary target on the outer membrane of Gram-negative bacteria
is a prerequisite of a purposeful design of novel polymyxin-like
lipopeptides.
Keywords: polymyxin, lipopolysaccharide, lipid A, docking, molecular modeling.
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10-16
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ÏÅÐÑÏÅÊÒÈÂÛ
ÑÎÇÄÀÍÈß ÏÐÎÒÈÂÎÄÈÀÁÅÒÈ×ÅÑÊÈÕ ÏÐÅÏÀÐÀÒΠÍÀ ÎÑÍÎÂÅ ÏÎËÈÔÅÍÎËÎÂ:
ÌÅÕÀÍÈÇÌÛ ÃÈÏÎÃËÈÊÅÌÈ×ÅÑÊÎÃÎ ÄÅÉÑÒÂÈß È ÔÀÐÌÀÊÎÊÈÍÅÒÈÊÀ
Ðóáàí Å. À., Êîëèñíûê Ò. Å., Ñëèï÷åíêî Ã. Ä.
PROSPECTS FOR DEVELOPMENT OF ANTIDIABETIC POLYPHENOL-BASED DRUGS: MECHANISMS OF HYPOGLYCEMIC ACTION AND PHARMACOKINETICS
Ruban E. A., Kolisnyk T. Ye., Slipchenko G. D.
Introduction.
Diabetes mellitus is one of the most serious chronic diseases and
considered to be non-infectious epidemic worldwide. Persistent
hyperglycemia is a major hallmark of diabetes and risk factor for the
development of its complications. Therefore, the main therapeutic goal
in the treatment of diabetes is to reduce the elevated blood glucose
level. Unfortunately, management of diabetes without any side effects
is still a challenge to the modern medicine and pharmacy. Among
potential alternatives to synthetic antidiabetic drugs plant
polyphenols are very promising. However, polyphenol efficiency in
diabetes is determined by their chemical structure and hence the
affinity to a certain molecular targets in body tissues. Moreover, the
bioavailability and other pharmacokinetic parameters of different
individual substances may also vary significantly. In this context the
present paper is devoted to the analysis of the available data on the
hypoglycemic mechanisms and pharmacokinetics of various individual
polyphenolic compounds in order to provide the necessary
biopharmaceutical requirements in the development of a new blood
glucose-lowering drug. Materials and methods. A systematic literature
search of Pubmed, EMBASE and other databases with no language
restrictions was performed until to the end of August 2015. The
following terms were used: polyphenols, diabetes mellitus, hypoglycemic
action, pharmacokinetics and bioavailability of polyphenols. Results and discussion.
According to available experimental data various polyphenols may
influence carbohydrate metabolism at many levels. The mechanisms by
which plant polyphenols exert their hypoglycemic action are mediated
primarily by their ability to directly bind to target proteins (or
peptides) and include inhibition of carbohydrate digestion and glucose
absorption in the intestine, stimulation of insulin secretion from the
pancreatic β-cells, modulation of glucose release from the liver,
activation of insulin receptors and glucose uptake in the
insulin-sensitive tissues. On the other hand, most polyphenols are
characterized by low bioavailability mostly due to intensive
metabolism. Thus absorption of such polyphenols as anthocyanins,
phenolcarboxylic acids and some others appears low, but it is supposed
that it could have been underestimated because not all metabolites
might have been considered. Besides the absorption rate of these
compounds is very rapid and may take place already in stomach. In
contrary, rutin and other quercetin glycosides are absorbed only after
release of the aglycones by the intestinal microflora. The elimination
half-lives of most polyphenols tend to be short, especially in the case
of anthocyanins. However, some polyphenolic compounds such as quercetin
glycosides may have longer half-lives, and even accumulate in plasma
with repeated ingestion. Conclusions.
Polyphenols have unique therapeutic potential in the treatment of
diabetes mellitus. Nevertheless, the possibility to use polyphenols as
hypoglycemic agents in clinical practice is limited by their low
bioavailability. Taking into account information reported in the
literature on the hypoglycemic mechanisms and pharmacokinetics of
polyphenols, promising method of increasing their bioavailability is
the development of prolonged-release dosage forms based on polyphenol
substances. This approach would extend residence time of polyphenols in
the small intestine – the main site of hypoglycemic action in their
intact, non-metabolized form, and will help maintain a constant
concentration of active substances in the blood plasma, the target
organs and tissues.
Keywords: polyphenolic compounds, diabetes mellitus, hypoglycemic action, pharmacokinetics, bioavailability
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17-24 |
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ÎÑÎÁËÈÂÎÑÒ² ²ÌÓÍÍί ²ÄÏβIJ ÏÐÈ ÃÐÈÏÎÇÍ²É ²ÍÔÅÊÖ²¯ ÒÀ ÏÅÐÑÏÅÊÒÈÂÈ ÑÒÂÎÐÅÍÍß ÂÀÊÖÈÍÍÈÕ ÏÐÅÏÀÐÀÒ²Â
Äàâèäîâà Ò. Â.
FEATURES OF THE IMMUNE RESPONSE DURING INFECTION AND PROSPECTS FOR THE VACCINES CREATION
Davidova T.V.
The
influenza virus belongs to the family Orthomyxoviridae and is a major
cause of respiratory infections in humans. Each year, influenza viruses
cause, according to experts, 3-5 million severe course of the disease
and 250 000-500 000 deaths. Influenza A viruses are divided into
serotypes based on their surface glycoproteins - known currently 17
subtypes of HA and NA subtypes ten. Upon infection with an influenza
virus, both innate and adaptive immune responses are inducing. In
recent years the annual seasonal epidemics were causing strains of the
virus A (H1N1 and H3N2) and virus B. This may be due to their ability
to be unrecognizable virus specific antibodies due to antigenic drift
(Figure 1). Seasonal flu vaccine, to be effective, must be updated
almost annually, according to new epidemic strains. In this work will
discuss various strategies used by influenza viruses to evade
innate immune responses and recognition by components of the humoral
and cellular immune response, which consequently may result in reduced
clearing of the virus and virus-infected cells.The primary targets for
influenza viruses are the epithelial cells that line the respiratory
tract and which initiate an antiviral immune response upon detection of
the virus. The first line of defense is formed by the innate immune
system, which is quick but lacks specificity and memory. Innate
immunity is formed by physical barriers and innate cellular immune
responses. Here, we outline several of the innate defense mechanisms
directed against influenza infections. During homeostasis, alveolar
macrophages exhibit a relatively quiescent state, producing only low
levels of cytokines, and suppress the induction of innate and adaptive
immunity. Activated macrophages enhance their pro-inflammatory cytokine
response, including IL-6 and TNF-α. Alveolar macrophages have a direct
role in limiting viral spread by phagocytosis of apoptotic infected
cells and by phagocyte-mediated opsonophagocytosis of influenza virus
particles. They are also involved in regulating the adaptive immune
response. The second line of defense against influenza is the adaptive
immune response. This highly specific response is relatively slow upon
first encounter with a pathogen. The adaptive immune response consists
of humoral (virus-specific antibodies) and cellular (virus-specific
CD4+ and CD8+ T cells) immunity. Influenza virus infection
induces the production of influenza virus-specific antibodies by B
cells. Antibodies directed to the viral HA and NA correlate with
protective immunity.Immune pressure on influenza viruses forces them to
adopt strategies to evade immunity.Various mechanisms contribute to
immune evasion of influenza viruses from the humoral immune response.
Due to the lack of proofreading activity, the transcription of viral
RNA by the viral RNA polymerase is error prone and results in
mis-incorporation of nucleotides. Under the selective pressure of
antibodies that are present in the human population, induced after
influenza virus infections and/or vaccination, variants are positively
selected from the quasi species that have accumulated amino acid
substitutions in the antigenic sites of HA that are recognized by
virus-neutralizing antibodies. This phenomenon is known as antigenic
drift. Introduction of influenza viruses of a novel antigenically
distinct subtype into the human population is known as antigenic shift
and may cause a pandemic outbreak, since neutralizing antibodies
against the new virus strain are absent in the population at
large.Introduction of antigenically distinct viruses can occur after
zoonotic transmission. However, in most cases, pandemics were caused by
viruses that had exchanged gene segments between human and avian or
swine influenza viruses. Currently used seasonal influenza vaccines are
predominantly inactivated vaccine preparations. Development of vaccines
that induce broad range of antibodies and preferably long
heterosubtypic CTL response is desirable. Yes viral proteins such as
the NP and M1 highly conservative, they are likely targets for the
induction of cross-reactive T cells. This requires effective delivery
of viral proteins in the cytosol. Several vaccine candidates cytosolic
delivery is currently being investigated, including DNA vaccines,
recombinant viral vectors, Iscoms and virosomal. They have already
passed clinical trials. In addition, the induction of cross-reactive
antibodies has attracted attention in recent years, antibodies directed
against conserved regions of molecules on the stem are of particular
interest. Unlike subtype specific antibodies induced against the main
round of the AB, these stem HA-specific antibodies capable of
neutralizing a broad activity against a large number of subtypes of
influenza virus. In addition, ectodomen M2 highly conserved
protein and antibodies induced against this region are able to create
protection against infection. Thus, vaccines that induce both
humoral and cell-mediated immune responses aimed at conserved areas of
virus, in addition to the strain-specific antibodies can afford.
Protective immunity to many different flu viruses, including new
variants and subtypes drift. Knowing the complexity of the interaction
between the immune system and variable pathogen such as influenza
virus, has increased significantly in recent years. This can help
reduce both morbidity and mortality, through the creation of effective
seasonal vaccine, but there are still gaps in understanding, providing
opportunities for improvement and developing a more widely-protecting
vaccines. Induction STL-reactions with the same
epitope can be a way to create fully protecting vaccines. Current
research also focuses on the responses of cross antibodies directed to
a more conservative regions of the surface proteins. Together, these
will create new ways to confront the changing nature of influenza
virus, and subsequently be able to protect even the emergence of new
pandemic strains.
Keywords: influenza; evasion; innate immunity; adaptive immunity
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25-39
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ÅÊÑÏÅÐÈÌÅÍÒÀËÜͲ ÐÎÁÎÒÈ
(EXPERIMENTAL STUDY)
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ÑÎÑÒÎßÍÈÅ ÈÌÌÓÍÈÒÅÒÀ ÁÎËÜÍÛÕ ÕÐÎÍÈ×ÅÑÊÈÌ ÔÀÐÈÍÃÈÒÎÌ ÍÀ ÔÎÍÅ ÑÒÀÔÈËÎÊÎÊÊÎÍÎÑÈÒÅËÜÑÒÂÀ
Îãíèâåíêî Å. Â., Ïîïîâ Í. Í.
STATUS OF THE IMMUNITY OF PATIENTS WITH CHRONIC STAPHYLOCOCCUS’S PHARYNGITIS
Ognivenko E.V., Popov N. N.
Immunomodulatory
and immunostimulatory drugs present the greatest interest in treating
such patients and the rehabilitation of their immune system. Today, a
wide clinical application has Polioksidonium (Petrovax Pharm), which
has a wide range of influence on the immune system. There are studies
proving the effectiveness of Polioksidonium treatment of inflammatory
diseases of the nasal cavity and paranasal sinuses, chronic
inflammatory process in pharyngeal plexus. Material and methods. We
observed 62 patients on chronic staphylococcus’s pharyngitis in age
from 21 to 45 years, who were treated in the communal health
institution "Kharkiv city hospital ¹ 6". The first group (basic group)
amounted to 32 patients, together with anti-inflammatory therapy
(topical Decatilen 1 tablet 3 times daily for 7 days, physiotherapy for
5 days, was obtained Polioksidonium (12 mg every 24 hours per os for 10
days). The second group (control group) consisted of 30 patients who
received similar therapy without Polioksidonium. Clinical,
microbiological, immunological studies were performed before the
treatment, at 7 and 30 days after the end of the therapy. As indicators
of standards the results of 30 healthy individuals were used.
Immunological studies included the determination of the phagocytic
activity of neutrophils, their biocidity, opsonizing properties serum
titers of antibodies to the causative infectious agents, concentrations
of the major classes of immunoglobulins in saliva and serum. Results
and discussion. Before the treatment the patients showed complaints to
pain and irritation in a throat, feeling of a foreign matter in a
throat, dry cough and low-grade fever. Microbiological
examination of oropharyngeal secretions ChFh patients revealed a 34 %
S. pneumonia, at 24 % - S.aureus. Microbial associations were sown in
45 % of the cases. Immunological studies have shown that patients with
chronic staphylococcus’s pharyngitis of increasing concentrations of
IgG and mIgA and reducing values sIgA and lysozyme in oropharyngeal
secretions. It has also been found that digesting and absorbing
capability of bacterial particles neutrophils peripheral blood of
patients with ChPh lower than that of healthy individuals. Low
phagocytic ability of cell for patients with HF was observed in respect
of opsonized autoserum bacteria. In all patients of the main group and
the comparison group revealed a significant increase in antibody titer
to bacterial etiological factors and common antigenic determinative
(CAD) bacteria. High antibody titers were detected for almost all
microbes studied. The clinical observations have shown that under the
influence of Polioksidonium patients with chronic staphylococcus’s
pharyngitis on the 7th day after the treatment has shown a marked
decrease or complete disappearance of the major clinical symptoms. All
patients receiving Polioksidonium markedly improved overall health. The
instrumental examination the 65% of the patients of the main group and
the 70% of comparison group on the 7th day after the end of treatment
were observed with swelling and hypertrophy of the mucous membrane of
the posterior pharyngeal wall. On the 30th day after the end of
treatment, 71.8% of the study group patients were without complaints
(in the comparison group - 30 % of patients), and their clinical status
was characterized by the norm. Pharyngoscope at 22 % of the patients of
the main group and at 66% of the patients of the comparison group
experienced a slight hypertrophy of the mucous membrane of the
posterior pharyngeal wall. Microbiological examination of the main
group found in 15,7 % of cases the ðresent of pathogens, marked
decrease in microbial associations without the appearance of fungal
flora and 68% of the pathogenic microflora. In the control group half
of the patients (52 %) was sown pathogenic flora, a slight decrease in
the amount of microbial associations. Under the effect of
Polioksidonium the increase of the secretory IgA and serum
immunoglobulins, and lysozyme occurred in the content of oropharyngeal
secretions. On the 7th day after the treatment the patients receiving
Polioksidonium, had a significant increase in the phagocytic activity
of neutrophils and biocide effect, and increase serum opsonizing
properties. The autoserum increased both in the absorption capacity of
neutrophils and their biocidal effect. Full restoration of the
functional activity of phagocytic cells in these patients occurred by
30 days after treatment. To compare the group of patients we have
studied that the properties of cells and serum opsonizing properties
recovering very slowly and by 30 days were significantly different from
the norm. On the 7th and 30th days after the end of the treatment the
patients who received Polioksidonium, had an increase in antibody titer
to infectious etiologic pathogens and IgG antibodies to CAD bacteria.
The patients who did not receive Polioksidonium, such dynamic
improvements in humoral immunity was observed. By 30th days after the
treatment the patients in a comparison group the antibody titer to
infectious etiologic pathogens and their affinity were not
significantly changed compared to their values before treatment. The
monitoring of patients during the year showed that the main group was
not observed recurrence ChFh. They are much less likely than the
comparison group was ill patients with acute respiratory infections,
which are mild and not accompanied by complications. In the study group
acute respiratory infections 2-3 times a year have been reported in 17
% of patients. Patients with ChFh comparison group relapses occurred in
40% of cases, of which 25% of patients - 2-3 times a year, 70% of
patients 2-3 times a year suffered acute respiratory illness, which in
23,8% of cases complicated by acute bronchitis, 15% - by acute
sinusitis. Conclusions: 1. Polioksidonium stimulates the increase
of phagocytic and biocidal activity of white blood cells, the
production of high affinity antibodies antimicrobial, increased serum
opsonin properties. 2. Using polyoxidonium decrease the manifestations
of pain and intoxication. 3. Polioksidonium has a positive effect both
on the clinical course of chronic pharyngitis and prevention of its
recurrence.
Keywords: Chronic pharyngitis, Polioksidonium, local immunity, oropharyngeal secretions.
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40-45
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²ÌÓÍͲ ÏÎÐÓØÅÍÍß Ó ÏÀÖ²ªÍÒ²Â Ç ÃÅÐÏÅÒÈ×ÍÎÞ ²ÍÔÅÊÖ²ªÞ ÐÎÒÎÃËÎÒÊÈ ÒÀ ¯Õ ÊÎÐÅÊÖ²ß
Ìàêàðåâè÷ Â.À., Êîëÿäà Ò.²., Òóïîò³ëîâ Î.Â., Êîëÿäà Î.Ì.
IMMUNE DISORDERS AND THEIR CORRECTION IN PATIENTS WITH HERPES INFECTION OF THE OROPHARYNX
Makarevich V.A., Kolyada T.I. Tupotilov O.V. Kolyada Î.Ì.
Recurrent
herpes infection of the oropharynx caused by herpes simplex virus type
I and II occupies an important place among the infectious diseases of
the oral cavity. The mechanisms which lead to the development of
recurrence of the disease are studied insufficiently, and this limits
the clinical possibilities of prophylactics. The frequency of the
manifestations of herpetic infection and their intensity depend on the
state of the immune system, in particular, the nature and level of
pathogenic disorders of immune response. The immunotherapy requires a
personalized approach taking into account the variability of
“sensitivity” of immune index of the patient for potential therapeutic
effects. The aim of the study was to justify the treatment of immune
disorders and timeous prevention of recurrence, and also complications
that occur when infection activates during the long-term viral
persistence, based on definition of the characteristics of immune
disorders in patients with moderate and high recurrence rate of the
disease. Materials and methods. The study involved 86 patients aged 18
to 49 years with herpesviral gingivostomatitis and pharyngotonsillitis
and herpetic vesicular dermatitis. The control group consisted of 19
healthy people of comparable age. Patients with recurrent HSV infection
were divided into two groups. Group I was formed of patients with
moderate rates of relapse less than 4 times per year (n = 51), and
group II represented patients with high rates of relapse not less than
4 times a year (n=35). Lymphocyte subsets were defined by fluorescent
microscopy method using FITC-labeled monoclonal antibodies against
surface antigens CD3, CD4, CD8, CD16, CD22. Evaluation of functional
lymphocyte reactivity was performed by setting 5 mg/ml PHA-stimulated
RBTL. Phagocytic activity was defined by latex method. The NBT-test was
used to evaluate the metabolic capacity of phagocytes. The CIC in serum
were defined by method of selective 3.5% PEG precipitation of antigen
complexes. Complementary activity of blood serum was determined by 50%
hemolysis. The content of IFN-α and IFN-γ in supernatants was
determined by ELISA after 24 hours of PBMC incubation. Results and
discussion. The level of immunodeficiency disorders expression by
herpes infection depended on the frequency of relapses during the year.
A decrease of white blood cells number and relative content of
lymphocytes of about 1.6 times as compared to the control group was
observed in patients with a high relapse rate of recurrent HSV.
Herewith, the relative content of CD8 + lymphocytes was 1.4 times lower
than the control and 1.5 times lower than the same index in patients
with moderate recurrent HSV relapse rate, and reduction of RBTL
stimulation index was expressed stronger as compared with the control
group. Levels of serum IgA and IgG were increased 2.1 times and 1.7
times respectively relative to the control group. Concentration of CIC
in serum of patients in both groups exceeded the control which points
to the failure of elimination function. A significant decrease in
phagocytic activity in both groups in patients with recurrent HSV was
more expressed in the group with high frequency of relapses. Also the
decrease metabolic reserves were found in both groups of patients which
points to the phagocytic function disorder. Elevated level of
B-lymphocytes and increased level of immunoglobulins points to the
activation of humoral immunity. Activity of complement in patients with
recurrent HSV was significantly higher than controls (1.4 times) in the
group with high frequency of relapses. Expressed deficiency of IFN-α
compared with controls and patients with moderate frequency of relapses
were observed in the group of patients with a high relapse rate. The
level of spontaneous production of IFN-α was elevated 1.6 times
compared to control in patients with moderate frequency of recurrent
HSV relapses, and the level of spontaneous production of IFN-γ was
elevated 1.3 times compared to control group. In our previous studies
we have determined that the content of perforin-positive cytotoxic
lymphocytes in patients with a high frequency of disease relapses was
reduced 2 times relative to healthy donors. It was shown that
HSV-inactivated vaccine was able to elevate the content of
perforin-positive cytotoxic lymphocytes in patients with moderate and
high recurrence rate; herewith the largest index of influence of
HSV-antigen was defined in patients with a high frequency of relapses.
HSV-induced production of IFN-α increased 3 times in patients with
moderate frequency of relapses and it had no significant difference
from the control group, and in patients with a high relapse rate - 2.3
times. Reserve ability to synthesize IFN-γ in patients with moderate
frequency of HSV relapses remained at the level of index of almost
healthy donors, while the expressed reduction of stimulating action of
HSV antigen was observed in patients with a high relapse rate. We have
previously shown that the high recurrence rate of herpes infection
accompanies by reduction of levels of TNF-α, by increase of spontaneous
and induced production of IL-10 (in 76% patients), IL-12 and IL-1β by
monocyte cells (in 85% of cases). Herewith 67% of patients with a high
frequency of disease relapses had an imbalance of IL-12p70/IL-12p40,
and 43% of patients had increased production of IL-1 receptor
antagonist.It was determined that incubation of PBMCs with the addition
of recombinant IFN-α causes a significant increase of the production of
IFN-γ in patients with moderate relapse rate and also in the control
group (stimulation index 1.3). The level of interferon-induced IFN-γ
had no significant difference from the basal level in patients with a
high relapse rate. Regulatory effect of IFN-α manifested also as
increase of production of IL-1β, herewith the increase of the ratio
IL-1β/IL-1RA was noted in patients with a high relapse rate.
Stimulation of IFN-α in patients with a high relapse rate leads to 1.4
times increase of induced production of IL-10 by cells of monocyte
fractions of MNC. The influence of IFN-α on the relative content of
perforin-positive lymphocytes in patients with recurrence HSV was less
expressed as compared to HSV-vaccine. So the stimulation index in
patients with moderate relapse rate was 1.4, while patients with high
relapse rate had stimulation index 1.2. Thereby, the choice of
immunotherapeutic strategy of treatment of recurrent herpes infection
of the oropharynx requires the immunological test to clarify the HSV
pathogenesis type.
Keywords: recurrent herpes simplex infection, immunodeficiency, interferons, cytokines, cytotoxic lymphocytes, immunocorrection
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46-53
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Ç̲ÍÈ Ì²ÊÐÎÁ²ÎÒÈ ÏÎÐÎÆÍÈÍÈ ÒÎÂÑÒÎÃÎ ÊÈØÅ×ÍÈÊÓ Ó ÕÂÎÐÈÕ ÍÀ ²Ë-²ÍÔÅÊÖ²Þ
Ñàâ³íîâà Î.Ì., Ãîðîá÷åíêî Ê.Ì., ͳêîëàºâà Ë.Ã. , Áîëüøàêîâà Ã.Ì. , Ìàéñòàò Ò.Â., Ä'ÿ÷åíêî À.Ã.
THE CHANGES OF LARGE INTESTINE CAVITY’S MICROBIOTA IN PATIENTS WITH HIV INFECTION
Savinova O.M., Gorobchenko K.M., Nikolaeva L.G., Bolshakova G.M., Majstat T.V.,
Djachenko A.G.
Introduction.
Infections of the gastrointestinal tract are caused by a wide range of
fungi, viruses and bacteria. The great value has the ratio of
microorganisms. There are certain regularities in microecological
system of intestinal microflora. Thus, bifidous bacteria should be more
than lactobacterium; enterobacteria – more than enterococcus;
E.faecalis more than E.faecium. However, these differences should be at
least one or two orders of magnitude. An important indicator is the
ratio of enterobacteria and enterococcus. Material & methods. In
the paper were used following materials and methods:
bacteriological and statistical. The conditions of intestinal
microbiocenosis were evaluated according to the methodical instructions
about researches of fecal masses on dysbiosis and modern methods of
correction of intestinal dysbiosis that included identifying of
anaerobic and facultative anaerobic microorganisms, staphylococci,
enterococci, opportunistic enterobacteria, Proteus spp., Klebsiella
spp., P.aeruginosa, C.albicans. Bacterial cultures were identified by
standard techniques. Statistical analysis of the results was performed
by the standard method of determining the average value and its
standard deviation (M + m) and Student's t-test. The reliability of the
difference was evaluated at the level of probability p <0.05. The
experiments’ data were processed using software applications Microsoft
Excel 2003 and «Biostat-6". Results & discussion.
We studied the species and quantitative composition of intestinal
microflora of patients with HIV infection. All studied patients had
disrupted quantitative and qualitative composition of intestinal flora.
The changes were identified both anaerobic and aerobic microorganisms.
Abnormality in the components of microbiota was manifested by reduction
in the number of microbiota in the absence of conditionally pathogenic
microflora. In the first place by deficit of microorganisms were
bifidus bacteria. According to our research in 94.1+4.01% of cases
bifidus bacteria were found in the amount of <106 CFU/grams and in
5.9+4.04 % of cases they were founded in the amount of 107 CFU/grams.
The second place by importance is occupied by lactobacillus. In
comparison with the norm patients with HIV infection had this indicator
reduced and amounts to 88,2+5,53% in ≤105 CFU/grams and 11.8+5.53% in
106–107 CFU/grams. Bacteroides from the group of anaerobic bacteria was
not found in any patient. Among the group of aerobic bacteria leading
representative was E.ñoli, which belongs to the group of obligate
microflora. 19 patients had E.ñoli in the amount of ≤106 CFU/grams,
which made up to 56% while the norm is 108 CFU/grams of faeces. 56% of
patients with the 3rd clinical stage of HIV had simultaneous shortage
of bifidus bacteria, lactobacillus and Escherichia. Hemolytic
E.coli were detected in small amounts of 2.9 + 2.93%. The
results of microbiological studies of fecal on dysbacteriosis shown
that despite the decrease of bifidus bacteria over 40 times in patients
with the 2–3rd clinical stage of HIV infection conditionally pathogenic
microflora appeared in small amounts, such as hemolytic isolates of E.
coli 2.9 + 2.93%, S.aureus in amount of >104 CFU/grams in 11,8 +
5,53%, S.epidermidis in amount of > 105 CFU/grams in 32,4 + 8,03%,
C.albicans in amount of > 104 CFU/grams in 11,1 + 6,59% and 1
patient had Clostridium in amount > 103 CFU/grams, which was 3 +
2,93%. These data show that patients with HIV infection is detected at
the same time reducing number of anaerobic microflora (bifidus bacteria
and lactobacilli) and aerobic microflora, a leading representative of
which is intestinal E.coli irrespective of the clinical stage of the
disease. Association of these microorganisms is 56%. To this attach
indicators of reduction E.faecalis and E.faecium. Taking into account
the features of obligate microfloras’ functions (bifidus bacteria,
lactobacillus, E.coli), its lack has a negative impact on
microecological system of the human body and reduces immunomodulatory
effect on humoral and cellular immunity. So one of the issues which
will have a positive impact on the health of patients with HIV
infection is a normalization of obligate microflora deficit and
reducing of opportunistic microflora. The conducted researches point to
the need of microbiological analysis of fecal on dysbiosis for
the patients with HIV infection and depending on the revealed dysbiotic
changes making correction of microflora by biological agents. To
correct the number of anaerobic bacteria (bifidus bacteria,
lactobacillus) use of bacterial preparations is not enough for only one
month. It is necessary to continue taking of medicine for at least one
month under the control of microbiological studies. The positive
dynamics of the microflora of the large intestine points to changes
that may be found in the immune system of the person that takes
biological preparations. The close interaction between the microbiota
of intestinal canal and the immune system leads to the formation of
non–specific resistance of the organism. In this regard, the big
importance has a modulating effect of intestinal microflora on products
of cytokines, which are characterized by a wide range of biological
effects. Conclusion. 1.
Patients with HIV infection irrespective of the clinical stage of the
disease have deficit both anaerobic (bifidobacteria and lactobacilli)
and facultative anaerobic microorganisms. 2. E.coli is the leading
microorganism among the facultative anaerobic intestinal microflora,
its amount of has been reduced to levels <106 CFU / mL at 56% at the
patients. 3. Correction of patients' microflora by bacterial
agents showed that the number of E.coli already in a month have reached
the normal level in 100% of cases.
Key words: dysbiosis, HIV infection, intestinal discharge.
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ÅÊÑÏÅÐÈÌÅÍÒÀËÜÍÅ
ÂÈÇÍÀ×ÅÍÍß ÇÄÀÒÍÎÑÒ² ÄÎ Ï˲ÂÊÎÓÒÂÎÐÅÍÍß ÌÅÒÈÖÈ˲ÍÎÐÅÇÈÑÒÅÍÒÍÈÕ ÒÀ
ÌÅÒÈÖÈ˲ÍÎ×ÓÒËÈÂÈÕ ØÒÀ̲ ÑÒÀÔ²ËÎÊÎÊÓ
Âîðîíê³íà ².À., Äåðêà÷ Ñ.À., Êðèëîâà ².À., Ãàáèøåâà Ë.Ñ.
EXPERIMENTAL STUDY OF BIOFILM-FORMING ABILITY TO METHICILLIN-RESISTANT AND METHICILLIN-SUSCEPTIBLE STAPHYLOCOCCUS STRAINS.
Voronkina I.A., Derkach S.A., Krilova I.A., Gabysheva L.S.
In
medicine there are many methods for studying of formation of biofilms.
Cultivation in dynamic systems allows to create the conditions as much
as possible close to that exist in the macroorganism. The majority of
experimental methods are based on building of static culture conditions
of microorganisms, most often for this purpose use 96-well plastic
microplates in various modifications. Using microplates even one
manufacturer leads to different results of the study, which is due to
physical and chemical characteristics. In addition, the formation of
biofilms using this method affects the composition of culture media
(micronutrient and electrolyte) and aeration degree.To quantify biofilm
formation in microplates researchers use the various wavelengths of the
photometer: 450 nm, 540 nm, 562 nm, 630 nm, 650 nm, etc. Thus, this
method is quite common, but its basic lack is that there are no
standards that would allow it to unify all laboratories.The aim of this
study was to optimize the test parameters to detect the ability of film
formation MRSA and MSSA strains. Materials and methods.
We used staphylococcus strains: S.aureus ATCC 25923, strains from the
laboratory of the museum with a certain in previous studies of
methicillin-sensitive and recently received S.aureus strains from
patients with purulent-inflammatory diseases of different localization.
Defining the properties of a biofilm forming staphylococcus culture
plates was performed by D. Christensen. Results and discussion.
In this experiment nutrient broth (HiMedia, India) was used with
different volume fractions ingredients: 2 % glucose, 6 and 7 % NaCl.
Microplates were incubated 4, 24, 48 and 72 hours at 35 ˚C. Optical
density of formed biofilm evaluated by the color intensity of spirit on
a photometer (StatFax 303 Plus). This method was studied
biofilm-forming properties of staphylococcus obtained from all 36
patients with community-acquired methicillin-resistant (MRSA) (n = 16)
and methicillin-susceptible (MSSA) (n = 20) strains. It is defined that
from 16 strains MRSA of 12,5 % had high ability to biofilms formation
of (OD 0,25-0,4 units), 3 strains of 18,75 % - an average (OD 0,12-0,
24 units), - have found 43,75 % weak ability to formation of biofilms
(OD <0,12 units), 25,0 % - a biofilm did not create in general (an
authentic difference in comparison with the control well it is not
revealed). It was defined that among 16 strains of MRSA 12.5 % had a
high ability to form biofilms (OD 0,25-0,4 units.), 18.75 % - average
(OD 0,12-0, 24 units), 43.75 % - found weak ability to form biofilms
(OD <0.12 units.) and 25.0% - do not create biofilm (significant
difference compared with control wells not detected). MSSA strains also
differ in their ability to form biofilms. 30.0 % of the isolates were
referred to the fact that not form a biofilm, 40.0 % - to strains with
weak ability to form a biofilm, 20.0 % - with the average, 10.0 % are
highly active biofilm formation. Conclusions.
1. The optimal parameters for the experimental determination of the
ability of biofilm formation of staphylococcus strains are: use of
nutrient broth with the bulk part of the 2 % glucose, incubating the
plates for 24-48 hours at 35 °C. The measurement of the optical density
(OD) of the resulting biofilms should be performed at wavelength of
630nm. 2. Experimental confirmation of a high ability to form biofilm
in the studied strains of staphylococci occurred in 10.0 % MSSA and
12.5 % - MRSA. 25.0 - 30.0 % of the strains were classified as not
forming a biofilm, and the rest - to the cultures with weak or medium
activity. 3. As a result of a comparative study of the ability to form
biofilms of circulating strains of community-acquired staphylococcus
significant difference between MRSA with methicillin-resistant
phenotype and MSSA strains not detected (χ2 > 0.05).
Keywords: staphylococcus, biofilm, the method of biofilm formation.
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ÀÍÒÈ̲ÊÐÎÁÍÀ ÀÊÒÈÂͲÑÒÜ ÅÔ²ÐÍÈÕ ÎË²É ÊÓËÜÒÈÂÎÂÀÍÈÕ ÏÐÅÄÑÒÀÂÍÈʲ ÐÎÄÈÍÈ LAMIACEAE JUSS. Øàíàéäà Ì.²., Ïîêðèøêî Î.Â.
ANTIMICROBIAL ACTIVITY OF ESSENTIAL OILS OF PLANTS BELONGING TO LAMIACEAE JUSS. FAMILY Shanayda M.I., Pokryshko O.V.
Introduction. One of the
important sources of therapeutic and prophylactic agents of modern
medicines are essential oils of medicinal plants. Essential oils are
the main group of biologically active substances of a number of plants
belonging to Lamiaceae Juss. Family. Antibacterial activity of
medicinal plants belonging to Lamiaceae Family many scientists
associated with containing of essential oils. In this regard,
considerable interest presents the comparative analysis of the
antimicrobial properties of essential oils of Lamiaceae Family
representatives. Material and methods.The
antimicrobial activity of essential oils of investigated plants was
studied with using in vitro condition. The essential oils derived from
the aerial parts of cultivated plants of Ocimum, Hyssopus,
Dracocephalum, Lophanthus, Monarda and Satureja genus harvested during
flowering period (in terms of Ternopil region). The antimicrobial
activity of essential oils studied plants was studied by serial
dilution method and disk diffusion assay. It has been applied on
standard microorganism test strains: Bacillus subtilis ATCC 6633,
Escherichia coli ATCC 25922, Staphylococcus aureus ATCC 6538,
Pseudomonas aeruginosa ATCC 9027 and Candida albicans ATCC 885-653. Results and discussion.
It was conducted a comparative study of the influence of some essential
oils of cultivated plants belonging to Lamiaceae family on
microorganisms in conditions in vitro. It was found that essential oils
of the studied plants were most effective in the maximum concentration
(1:10). Gram-positive cocci S. aureus and yeast C. albicans were the
most sensitive to influence of investigated essential oils. It
was analyzed the relationship of the biological activity with the
component composition of essential oils of plants. Essential oils of L.
anisatus, M. fistulosa and S. hortensis characterized by the dominance
of aromatic compounds and had shown stronger antimicrobial activity
than essential oils of other species. Therefore, essential oils of
these species are very promising source of antibacterial and antifungal
drugs, especially in the treatment and prevention of skin diseases,
upper respiratory tract inflammations and vaginitis. Conclusion.
Obtained results indicate promising further study of antibacterial
properties of essential oils of plants species L. anisatus, M.
fistulosa and S. hortensis (Lamiaceae family). It was discovered that
Gram-positive cocci, including S. aureus, and yeast Candida were the
most sensitive to these oils. Along with the prospect of creating
antimicrobial drugs which include essential oils of studied plants
these oils can be recommended as active ingredients of preservatives in
the composition of cosmetic products or dietary supplements also.
Keywords: Lamiaceae, essential oils, antimicrobial activity
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ÈÑÑËÅÄÎÂÀÍÈÅ ÝÊÎËÎÃÈ×ÅÑÊÈÕ ÎÑÎÁÅÍÍÎÑÒÅÉ ÂÎÇÁÓÄÈÒÅËÅÉ ÎÑÒÐÛÕ ÄÈÀÐÅÉÍÛÕ ÈÍÔÅÊÖÈÉ
Ìàëûø Í.Ã., Õîëîäèëî Å.Í., ×åìè÷ Í.Ä.
INVESTIGATION OF ECOLOGICAL FEATURES OF ACUTE DIARRHEAL INFECTION PATHOGENS
Malysh N.G., Holodilo E.N., Chemych N.D.
Introduction.
Microbiocenosis of human body also differs in extreme multicomponents
and diverse content of microflora representatives forming its
part. According to the biotype of bacterial contamination certain
inter-bacterial relations are formed, which is reflected in the
qualitative and quantitative characteristics of appropriate microbial
landscape. Analysis of numerous microbial association manifestations
allows evaluating changes in the pathogen properties influenced by
associative microbiota. Work objective - based on the study ecological
features of microorganisms isolated from intestine of patients with
acute intestinal infections and apparently healthy people, identify
potential risk factors for diarrheal infections. Materials & methods.
A retrospective epidemiological analysis of acute diarrheal infections
incidence was conducted during 2004-2013, using the statistics of the
Main Department of the State Sanitary and Epidemiological Service of
Ukraine in Sumy region. The intestinal microflora of 93 patients with
acute diarrheal infections and 60 persons of the control group
(apparently healthy people). As the result 130 bacterial cultures
were allocated. Permanence rate was used to estimate biocenosis.
Relationships between microbiocenosis members were investigated by
determining degree of bond conjunction in associations, using Jaccard
coefficient (g). Results & discussion.
In 2005-2014 acute diarrheal infection incidence rates of Sumy region
population were within 163.7 - 193.6 per 100 people without tendency to
decrease. Acute intestinal infections and food toxicoinfections caused
by opportunistic pathogens and viruses (p<0.05) dominated in
nosological structure. In 35.5 % of cases diarrheal infections were of
polyetiological nature. Noroviruses in associations with Candida
bacteriaand fungi most often occurred (p<0.05) in the intestinal
biotypes. Permanence rate of K. pneumonia, noroviruses, S. aureus, C.
albicans was the highest and was accordinly 23.6; 21.5; 19.4 and 19.4
%. C. albicans (88.9 %), noroviruses (80 %), S. thyphimurium (80 %), P.
aeruginosae (75 %), E. cloacae (75 %), S. aureus (72.2 %), K. pneumonia
(68.2 %), rotaviruses (66.7 %) had average and high associativity
coefficient values. The most part of associated causative agents
(p<0.05) had antagonistic relations. Stable bacterial associations
were formed between noroviruses and K. pneumonia, noroviruses and
S. typhimurium, noroviruses and P. aeruginosae (g respectively 36.4;
40.0; 40.0 %). C. albicans, K. pneumoniae, S. aureus, E. cloacae were
isolated from healthy people feces in diagnostically significant degree
of contamination 43.3 % of cases. The permanence coefficient rated
respectively 26.7; 20.0; 20.0; 3.3 %, as well as the associativity
coefficient was 90.0; 50.0; 83.3; 100 %. K. pneumonie and S. aureus
(g=42.9 %), S. aureus and C. albicans (g=33.3 %) cultures showed the
greatest strength of microbial relations. Conclusion.
In the process of microorganism co-existence different kinds of
relations are formed between them - competitive or cooperative, and as
the result the specific microsymbiocenosis is formed according to them.
Having established universal common factors of this microsymbiocenosis,
you can find ways to protect body from infection.
Keywords: acute diarrheal infections, microorganism association, ecological features.
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ÑÈÍÒÅÇ,
Ô²ÇÈÊÎ-ղ̲×Ͳ ÂËÀÑÒÈÂÎÑÒ² ÒÀ ÏÐÎÃÍÎÇ Á²ÎËÎò×Íί ÀÊÒÈÂÍÎÑÒ² ÍÎÂÈÕ
ÏÎÕ²ÄÍÈÕ 2-R1-N-(5-R)-1,3,4-Ò²ÀIJÀÇÎË-2-²Ë- ÁÅÍÇÎËÑÓËÜÔÎÍÀ̲IJÂ
Ñè÷ ².Â., Ïåðåõîäà Ë.Î., ªðüîì³íà Ç.Ã.
NEW DERIVATIVES OF 2-R1-N-(5-R)-1,3,4-THIADIAZOL-2-YL-BENZOLSULFONAMIDES: SYNTHESIS, PHYSICOCHEMICAL PROPERTIES AND BIOLOGICAL ACTIVITY PREDICTION Sych I.V., Perekhoda L.Î., ²åremina Z.G.
Introduction: The
analysis of modern literature, including overseas
one, showed that a lot of the scientific
researches is devoted to finding and creating biologically active
compounds on base 1,3,4-thiadiazole. Derivatives of 1,3,4-thiadiazole
are the large group of heterocyclic compounds with high rates of
antimicrobial, antituberculosis, antidiabetic, antineoplastic and
anticonvulsant activity. Material and methods: The
purpose of this study was the expansion of sulfone derivatives
substituted nitrogen-containing heterocyclic systems through the
synthesis of 2-R1-N (5-R)-1,3,4-thiadiazol-2-ilbenzolsulfonamides and
prediction their pharmacological activity for future planning
pharmacological screening. Synthesis of semi-products
2-amino-5-R-1,3,4-thiadiazoles was carried out by cyclization
thiosemicarbazide and substituted derivatives of carboxylic acids in
the presence of concentrated sulfuric acid. The synthesis oftarget
compounds 2-R1-N(5-R)-1,3,4-thiadiazol-2-ylbenzolsulfon-amides was
carried out by N-acylation of 2-amino-5R-1,3,4-thiadiazole substituted
benzolsul-fochlorides in the presence of anhydrous pyridine. The
reaction proceeds by the classic SN2-mechanism. The resulting compounds
are white crystalline substances, soluble in alcohol, chloroform and
acetone, difficult to dissolve in water. Yields of obtained compounds
was satisfactory (76-84%). The purity of the obtained compounds was
determined by TLC. The structure of the obtained compounds was proved
by elemental analysis, IR methods and 1H NMR spectroscopy.NMR 1H
spectra were recorded at Bruker WM spectrometer (200 MHz); solvent
DMSO-d6; chemical shifts were in ppm, internal standard (TMS
(tetramethylsilane)) was used. The prognosis of biological activity for
obtained compounds were carried out using the program PASS (Prediction
of Activity Spectra for Substances) in order to plan the further
pharmacological screening. The program PASS predicts more than 500
kinds of biological activity using the chemical structural formula and
helps to narrow the limits of experimental screening to identify
compounds-leaders. This program is used just to assess the affinity of
new compounds with known drugs, that is, to characterize their «drug
likeness». The calculation was conducted with the help of Internet
service that offers the software package of processing of listed
structures and calculation of properties. Results and discussion:
Synthesis of new potential biological active substances 2-R1-N
(5-R)-1,3,4-thiadiazol-2-ilbenzolsulfonamides has been carried out.
Finished products have been obtained by the interaction of
2-amino-5-(R)-1,3,4-thiadiazoles with corresponding substituted
benzolsulfochlorides in the presence of anhydrous pyridine. Target
products have been obtained with satisfactory yields. All semi-products
have two-proton signal of aminogroup in the area from 2.5 to 2.82
ppm in NMR1Í spectra. Signals of protons substituents
in the second position of the heterocycle (methyl, propyl, i-propyl,
butyl, sec-butyl) on corresponding areas of spectra is present in NMR1Í
spectra of the all initial compounds aswell. In contrast to the
spectrums of initial compounds signals of amino groups at the spectra
of the finished compounds are absent. All spectra of the target
products have the secondary amino group signal at 6.81 and 6.71 ppm
areas. The purity of the obtained compounds determined by TLC.
Prediction of biological activity derived substances was conducted
using a computer program PASS. Conclusion.
The group of 2-R1-N-(5-R)-1,3,4-thiadiazol-2-ylbenzolsulfonamides has
been synthesized by acylation of the corresponding 2-amino-5
-(R)-1,3,4-thiadiazoles. The structures of the synthesized compounds
have been proved by elemental analysis, IR and 1H NMR spectroscopy
data. All substances for which the PASS program prognosis was carried
out can show themselves as potential antidiabetic and antimycobacterial
drugs.
Keywords: 1,3,4-thiadiazole, synthesis, IR and 1H NMR spectroscopy, prognosis of biological activity.
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²ÄÍÎÂËÅÍÍß IN VIVO ÍÅÊÓËÜÒÓÐÀÁÅËÜÍί ÑÓÁÏÎÏÓËßÖ²¯ SALMONELLA ENTERICA
Þä³í ². Ï.
RECOVERY IN VIVO OF NONCULTURABLE SUBPOPULATION OF SALMONELLA ENTERICA
Yudin IP
Introduction.
As one of mesophilic, easily cultivated species of pathogenic bacteria,
Salmonella enterica transformed into viable but nonculturable (VNC)
state in response to environmental stresses, including action of
biocides. The cells in this state, preserve the integrity of membranes
and metabolism of some, but not detected by conventional methods of
cultivation. Some researchers suggest that the evolutionary
significance of this phenomenon is part of an adaptive response aimed
at long-term survival of bacteria in adverse conditions; others argue
that it is the result of stochastic cellular damage, in which
nonculturable cells are in a state of gradual death. In any case, the
phenomenon of existence VNC pathogens if they retain the ability to
restore its growth in vivo is a significant problem in medicine,
pharmaceutical, veterinary, food industry. VNC subpopulation of S.
enterica was obtained under action of ethanol. In this paper was
investigated in vivo resuscitation VNC S. enterica using
intraperitoneal injection of mice. Materials and methods.
Obtaining of stressful S. enterica populations. Bacteria were grown to
exponential phase in broth Luria–Bertani (LB). To 1.0 ml sample
suspension diluted to 1.5 × 106 cells/ml was added 1.0 ml of ethanol at
a concentration of 40 % (v/v). After exposure of 10 to 600 minutes in
the suspension were added 8.0 ml of phosphate buffered saline (FBS),
washed by centrifugation (4500 g for 5 minutes) and serially diluted at
a ratio of 1:10 (v/v) samples were stained with LIVE/DEAD BacLight
(produced by "Invitrogen", USA), filtrated on membrane filters for
fluorescence microscopy and parallel plated on LB agar cup to determine
colony-forming units (CFU) per ml. In vivo resuscitation VNC S.
enterica was made following way. Three groups of animals were
inoculated by intraperitoneal injection: 1) 103 culturable cells (0.1
ml suspension containing 104 CFU / ml); 2) 103 VNC cells (0.1 ml
suspension containing 104 cells / ml of nonculturable population); 3)
103 inactivated cells (pasteurization at 60 °C in 70 % ethanol, 30
min). Mice were observed daily for 14 days to register the death and
extraction of internal organs. Liver and spleen of dead mice were
removed and homogenized in 1.0 ml of FBS to detect restored Salmonella
cells by seeding on selective medium bismuth sulfite agar (BSA). Results and discussion.
In this study S. enterica cells in the exponential growth phase,
exposed to ethanol (final concentration 20 %) lost culturability within
60 minutes. After 50 minutes of exposure with ethanol culturability of
bacterial suspensions was outside evaluation cup method. At the same
time, the integrity of cell membranes was determined at 4 log10
cells/ml. Bacteria that become VNC state can restore culturability.
Thus, the state is reversible. Importantly, the resuscitation of VNC in
vitro, which is achieved by simply eliminating or VNC induction factor
(increasing the growth temperature, availability of nutrients) or more
complex conditions such as a combination of environmental and chemical
stimuli, does not reveal the full pathogenic potential of resuscitated
bacteria. Therefore, the process of Salmonella resuscitation in vivo,
we studied on infection model in mice. In experiments in vivo S.
enterica resuscitation death of test animals was observed in 13.3% (p
< 0.05) during the observation period, with 100% mortality in the
group of animals infected with a suspension of living culturable cells
Salmonella and 100% survival of the animals in the control group (who
received injections of inactivated bacteria). From the dead mice from
homogenates of internal organs plated recovered salmonella, which was
confirmed by their growth on BSA. Conclusions.
Under conditions in vivo recovery process VNC S. enterica cells can
occur, but with low intensity in healthy mice (at 13.3%, p <0.05).
However, these cells retain pathogenic potential and can represent a
danger if their underestimation. We can assume that the bacteria lose
their virulence in part, but manifest it in individuals with a weak immune response.
Key words: VNC, Salmonella, resuscitation.
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ÄÎÑ˲ÄÆÅÍÍß
ÀÍÒÈÁÀÊÒÅвÀËÜÍί ÀÊÒÈÂÍÎÑÒ²ÊÎÌÁ²ÍÎÂÀÍÈÕ ÏÐÅÏÀÐÀҲ ÍÀ Ê˲Ͳ×Ͳ
ØÒÀÌÈ̲ÊÐÎÎÐÃÀͲÇ̲Â, ÂÈIJËÅÍÈÕ Â²Ä ÕÂÎÐÈÕ ÇÁÀÊÒÅвÀËÜÍÈÌ
ÂÀòͲÒÎÌ Àñëàíÿí Ì.À., Áîáðèöüêà Ë.Î., Ñîëîíèíà Í. Ë., Îñîëîä÷åíêî Ò. Ï., Êó÷ìà ². Þ.
INVESTIGATION
OF ANTIMICROBIAL ACTIVITY COMBINED PREPARATIONS FOR CLINICAL STRAINS OF
MICROORGANISMS ISOLATED FROM PATIENTS WITH BACTERIAL VAGINIT Aslanian M. A., Bobritskaya L. O., Solonina N. L., Osolodchenko . T. P., Kuchma I. Y.
The problem of bacterial vaginit in some cases the
cause of severe infectious diseases genitalia of the fetus and newborn,
which can impair the health of future generations. It is noted that the
treatment of antibacterial agents observed numerous negative side
effects- reducing the biochemical activity of the intestinal
microflora, abuse microbiota, leading to the development of dysbiosis,
increasing the number of resistant strains of pathogens, the risk of
allergic reaction sand immunological disorders. A study was conducted
towards finding effective combinations of drugs from different
pharmacological groups means to create a combination of drugs. The aim
of the study was to develop and explore and Flamini combination of
miramistin combined medicines to treat bacterial vaginit. As a result
of studies in patients with bacterial vaginit pathological material was
isolated and identified 72 strains of microorganisms (Staphylococcus
spp, Streptococcus spp, Enterococcus spp, Escherichia coli, Haemophillu
sssp, Candida albican sand various strains of anaerobic
microorganisms. For the combined treatment of infectious and in
flammatory diseases (mixed infections) in humans the combined drugin
tablet form. All clinical strains of microorganisms isolated from
patients with bacterial vaginit were tested for sensitivity to the
combined preparation in tablet form with Flamini and miramistin. The
greatest sensitivity to the drugs found clinical strains of
microorganisms: Staphylococcu saureus, Staphylococcus epidermidis,
Peptococcus niger (diameter zone growth retardation is 25,5-23,5
mm). composition tablets number 1 (0.05 g Flamini,
miramistini 0.02 g), which was selected for further study
shows bacteriostatic effect against a wide range of microorganisms and
fungi Rod Candida. IPC for Staphylococcus sp was 20-25 pg / mL for
Streptococcus sp 35,0-40,0 mg / ml, for intestinal group 35,0-40,0 for
fungi 30,0 mg / ml unlike pills number 2 and number 3, where the IPC
was higher and amounted to 40,0 -55,0 mg / ml. Summing upthe results of
experimental studies capsules combined with the composition Flamini and
miramistin becon cluded that the most effective eagainst the different
clinical strains of microorganism sand fungi is a combination Flamini
0,05g, miramistin 0,02 (tablets number1), giving opportunity to develop
drug combinations for use in bacterial vaginit.
Key words: vaginitis, clinical isolates, miproorganisms, activity, antimicrobial drugs.
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ANTAGONISM OF A. VIRIDANS TO CONDITIONALLY - PATHOGENIC MICROFLORA OF THE NOSE AND OROPHARYNX OF CHILDREN WITH CARDIAC PATOLOGY
Stepansky D.O.
Introduction.
Search for harmless and simultaneously effective probiotics, which
could be successfully used for treatment and prevention of infectious
deseases, is currently important. A. viridans is of particular
interest, as it is representative of the normal microflora of human
with broad spectrum of antibacterial action. The use of this
microorganism has a number of advantages: the absence of side effects
on the body; high adhesive abilities; resistance to lysozyme in saliva;
the ability of use in patients, sensitized to antibiotics and
chemotherapeutic drugs; stimulation effects on the human immune system.
Material and methods. The
purpose of the study was to investigate the antagonism of A. viridans ¹
167 and autostrains of aerococcuses, isolated at patients, to
conditionally - pathogenic microflora of the nose and oropharynx of
children with cardiac patology. At the first stage of the study the
microflora of the of the nose and oropharynx of 2 investigated
categories was examined – 40 children 4-14 years with cardiac patology
and 40 healthy children 4-5 years old. The second stage of work
was to study the effect of A. viridans on the explored strains. Results and discussion. A.
viridans manifests the antagonism to all studied strains of
gram-positive and gram-negative microorganisms, except C. albisans. A.
viridans antagonistic activity to staphylococci (10 + 3 mm) and
streptococci (10 + 2mm) is at the approximately same level. It is
interesting to compare the antagonism of aerococcuses to clinical
isolates of S. pyogenes and similar strains from carriers (healthy
children category). Impact of aerococcuses on P. mirabilis strain
appeared at the highest level. Autosimbionts of A. viridans, isolated
from healthy children, are more antagonistic to CPM strains, isolated
from these children, than autostrains of A. viridans, isolated from
children with with cardiac patology, and higher than the museum strain
of A. viridans ¹ 167 antagonism, and do not affect the growth of
microorganisms of Candida kind. Severe antagonistic activity of the A.
viridans ¹ 167 strain and autostrains of A. viridans to isolated
conditionally patogenetic strains was discovered: S. pyogenes, S.
pneumoniae, S. aureus, K. pneumoniae, P. mirabilis. Antagonism of A.
viridans ¹ 167 to C. albicans was not found. Conclusion.
The recieved data can serve as a basis for further study of the strain
of A. viridans ¹ 167 and aerococcuses’ autostrains usage for correction
of fauces and nasopharynx microbiocenosis and also for the complex
treatment and preventive actions in case of cardiac pathology.
Key words: Aerococcus viridans, probiotics, autostrains of aerococcuses, children.
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90-93 |
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