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Editorial Board |
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Experimental works |
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Determination of microbiological purity of rectal
suppositories with
diosmine and hesperidine Borko Ye. A. , Kovalevska I.V. , Osolodchenko T.P. Introduction. The use of conservative methods of treatment, in terms of
increasing the
number of patients with anorectal diseases, is becoming increasingly
important
in modern realities. Proctological pathologies are becoming more
characteristic
for people of working age, and can affect not only the physical and
mental
health of an individual, but also the future reproductive state of the
nation
as a whole. , It is advisable to use dosage forms that will effect on
several
factors in the pathogenesis, while showing resorptive and local
effects. As an
example, of such a dosage form (DF) are rectal suppositories, which
according
to a complex of structural-mechanical and biopharmaceutical factors is
the
optimal DF for the release of active substances on the place of the
pathological process. An important stage in the formulation of rectal
suppositories it isn`t only the choice of active substances, but also
quality
control of ready-made DF. The SPhU of Ukraine regulates the quality
indicator
"microbiological purity" for suppositories as a critical stage of
control, which means the necessity for a full range of studies to
identify
possible of microbial contamination in the DF. Material
& Methods. We have selected 3 samples of rectal
suppositories with diosmin and hesperidin for microbiological studies.
In order
to substantiate the stability during storage, we have proposed to use
samples
with different durations from the date of manufacture (1- just made; 2-
2 weeks
storage; 3- 6 months storage). The testing of suppository samples for
microbiological
purity was carried out using with the Chistovich medium, blood
agar-based soybean
casein digest agar and Endo medium. As a preliminary preparation for
the study
of microbiological purity, tests were performing for compliance with
the growth
qualities of nutrient mediums. The definitions of microscopic study
were
carried out a microscopic method using a Konus Academi Microscope of
Italian
production with a DLT-Cam Basic 2MP camera. The test of microbiological
purity
was conducted with methods direct seeded on liquid nutrient mediums.
The
definitions of quantity of viable cells of microorganisms and fungus
for
methods of deep seeding have been conducting with adding sample in
quantity 0.1
in agar medium. Results &
discussion. The results of the study of growth
properties of nutrient mediums was shown, that all cultures of
microorganisms
met to the taxonomic designation of the strain. The obtained results
have shown
that after 14 days of incubation of suppositories samples on the Saburo
medium
- the fungus haven't growth. The results of researches
samples with soybean casein digest broth and thyoglycollate medium
in the quantity 1.0 and 0.1 haven`t shown the growth of microorganisms.
Registration of growth of the microorganisms has been shown in research
of
samples in the quantity of 0.01. The microorganisms that have been
detected
during the studies by morphology of colonies and biological properties
belong
to the genus Bacillus sp. According to the research results by the
method of
direct seeding on cups the growth of fungus has been absented in all
suppositories samples. Conclusion. Taking
into account the obtained data we was indicated that viable fungal
cells
weren`t detected in the suppositories samples. The quantity of
microorganisms
was lower by 103 CFU/ml that meets the requirements of SPhU.
Microorganisms that have been detected during the studies belong to the
genus
Bacillus sp. The strains of S. aureus, P. àeruginosa and
representatives of the
genus Enterobacteriacea sp. haven`t been detected. Suppositories
samples that
have been determinations on indicator of microbiological purity, meets
the
requirements of SPhU. Keywords: microbiological purity, rectal
suppositories, diosmine, hesperidine
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9-12 |
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Determination of the optimal extractant for the
extraction of biologically active substances of Sophora
Flower–buds Kriukova A., Bezruk I., Konovalenko I. Introduction. According to the market
products
based on Sophora japonica L., extraction of biologically active
substances,
flavonoids in particular, is carried out with ethanol in a
concentration
50–70%. However, in recent years, in literature data have appeared the
information that the use of water–ethanol solutions is not effective
for the
extraction of biologically active substances and contributes to the
release of
a large amount of ballast substances. An alternative is to use
surfactant–based
extractants. In relation to Sophora Flower–buds, these questions have
not been
investigated, which determines the relevance of research on the
selection of
the optimal extractant. Material &
methods. The object of the research
is Sophora Flower–buds, harvested in 2018 during the budding period.
The
definition of quality indicators was carried out in accordance with the
requirements of the State Pharmacopoeia of Ukraine. Results
& discussion. Determination
of Sophora Flower–buds technological parameters was conducted:
the
degree of grinding of herbal materials, specific mass, bulk density,
bulk
weight, porosity, permeability and free volume of the layer. The obtained data on the technological properties of the
herbal material
used to develop an optimal method for extracting, predicting and
standardization of the quality of extracts. Studied one of quality
indicators,
that indicates a rationally selected extractant is “Determination of
dry
residue of extracts”. Also was carried out research a comparative
evaluation of
the flavonoids content in the extracts obtained. Quantitative
determination of
the sum of flavonoids was performed by spectrophotometric method
according to
the SPhU 2.1 method «Sophora Flower–buds». The data obtained indicate
that the
highest values of dry residue and maximum amount of flavonoids were
obtained
during the extraction for two to three hours using ethanol 50 % and
sodium
lauryl sulfate solution 2.5 % as the extractant. Content flavonoid
compounds of
different chemical composition Sophora Flower–buds can affect
biological
activity. Therefore, for a more objective evaluation of the extracting
ability
of the proposed extractants were conducted studies of antiradical
activity.
Research of the antiradical activity of Sophora Flower–buds showed that
the
maximum value of TEAC is exhibited by such agents as ethanol 50 % and
sodium
lauryl sulfate solution 2.5 %. The data obtained confirm that the
antiradical
activity depends on the quantitative content of flavonoids in the
studied
samples of Sophora Flower–buds. Conclusion. The research of possibility of
using surfactants for the extraction of biologically active substances
from
Sophora Flower–buds was conducted. It is found that the best extractive
ability
to Sophora Flower–buds flavonoids have extractants: ethanol 50 % and
sodium
lauryl sulfate solution 2.5 %, which have virtually the same quality
indicators: dry residue 8.81±0.37 and 8.54±0.45; the content of the sum
of
flavonoids 8.75±0.01 è 8.43±0.03. The study of antiradical activity
in Sophora Flower–buds extracts was conducted for the first time.
Maximum
values are obtained in extracts by extraction over two hours using
ethanol 50 %
sodium lauryl sulfate solution 2.5 %. The data presented can be used in
research
on the development of Sophora Flower–buds herbal medicinal products in
the
various dosage form. Keywords: Sophora Flower–buds, extractant, biologically active
substances
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13-16 |
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Testing of
antimicrobial activity of preservatives for dental gels development Maslii Yu.
S., Ruban O. A, Kaliuzhnaia
O. S., Khokhlenkova N. V Introduction. Given the widespread prevalence of oral diseases,
especially
pathologies of periodontal tissues and mucous membranes, among the
population
of different ages and the needs of the pharmaceutical market in topical
drugs
for their treatment, the development of new domestic dental drugs is
relevant.
The most common topical dosage form in dental practice is gels, which
are
characterized by good distribution on the tissues of the oral cavity,
prolonging effect and high bioavailability. At
the Industrial Technology of Drugs Department of National University of
Pharmacy, two dental gels are being developed under the name
"Cholident" and "Lysostom". Previous studies showed
that freshly prepared gels satisfied the requirements of SPhU for
microbiological purity. However, contamination of the samples by
test-microorganisms caused their intensive growth, which proved
necessity for
introduction of antimicrobial preservatives into the gels. These
adjuvants prolong the shelf life of
pharmaceutical products, increase their resistance to spoilage after
opening
the package, especially in the case of using multi-dose containers,
and,
accordingly, prevent infection of the patient. The aim of this work was selection of
effective
antimicrobial preservatives in rational concentrations for the
composition of
dental gels "Cholident" and "Lysostom". Materials
& methods. The
following ingredients
were selected as preservatives in the gels are being developed: benzoic
acid, sodium benzoate, sorbic acid, potassium sorbate, methyl
parahydroxybenzoate (nipagin), propyl parahydroxybenzoate (nipazol). The
concentration of preservatives for all gel samples
was 0.1 % and 0.2 %. They have much lower toxicity than
others, are
harmless to humans even in large quantities and are permitted for the
preservation of food and pharmaceutical products. The selection of
preservative
for the composition of dental gels "Cholident" and
"Lysostom" was based on the study of its antimicrobial activity.
Preservatives effectiveness tests were carried out according to the
method of
SPhU 2.3, Ch. 5.1.3. According
to the requirements of SPhU, the sterility
test of culture medium and solvent: growth properties of culture medium
(soy-casein culture medium – for bacterial cells growth and
Saburo-dextrose
medium without antibiotics – for fungal cells) and suitability test of
methods
for determining the total number of cells were performed. Results
& discussion. Tests have shown that for dental gel "Lysostom"
with preservatives nipagin, sodium benzoate, potassium sorbate, it
could be
noted that the obtained data satisfy the requirements of SPhU for oral
drugs.
Among the preservatives listed above, sodium benzoate at a
concentration of
0.2 % had the highest antimicrobial activity. The results of
antimicrobial
preservatives efficacy determination in the composition of the dental
gel "Cholident"
proved SPhU eligibility
for all samples. The combination of nipagin : nipazol had the
highest
antimicrobial activity against all test-microorganisms, and, taking
into
account the requirements of economy and safety, their use at a
concentration of
0.1 % will be sufficient. Conclusion. Thus, tests of the
antimicrobial preservatives effectiveness in the experimental samples
of dental
gels "Lysostom" and "Cholident"
proved that all the studied preservatives, namely sodium benzoate,
potassium
sorbate, nipagin and combination nipagin with nipazol, benzoic acid and
sorbic
acid, had high antimicrobial efficacy and met the requirements of SPhU
for oral
drugs. Taking into account a slightly higher antimicrobial activity,
the sodium
benzoate at a concentration 0.2 % was selected as the most
acceptable
preservative for the gel "Lysostom". The combination
nipagin : nipazol (3:1) was chosen as the most promising
preservative
for the gel "Cholident",
which
provided at a concentration 0.1 % stronger than other
preservatives
antimicrobial action. Key words: dental gel, preservative, microbiological studies, antimicrobial activity
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17-23 |
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Using the HACCP method in quality risk management in the
production of
oromucosal gel Orlenko D. S., Yakovenko V. K. Introduction. With the adoption in the EU of the regulatory document of
the European Medicines
Agency (European Medicines Agency) EMA / INS / GMP / 79766/2011
“Quality Risk
Management (ICH Q9)” separate guidance was issued in Ukraine in 2011 -
Instruction ST-N MOH 42-4.2: 2011 “Medicines. Quality Risk Management
(ICH
Q9)”. Pharmaceutical industry and regulatory bodies
professionals can assess and manage risk using recognized risk
management tools
and / or internal techniques (e.g., standard working methods). Application of general principles and approaches of
the Guideline “Medicines. Quality Risk Management (ICH Q9)”at the stage
of
pharmaceutical development, using appropriate risk management tools
both in
general to the manufacture of medicines and to individual processes is
an
effective measure of quality assurance for the developed medicinal
product. The
technology of the combined dental gel was developed taking into account
its
properties as a dispersed system, as well as the properties of the
active and
auxiliary substances that are part of it. In order to manage quality
risks
effectively, it is necessary to have data about the sustainability of
the
process. Critical process parameters that need to be managed or
monitored to
ensure the required quality of the drug should be identified and
specified. Materials and
methods of the research. The subject of the
study was the technology and technological scheme of the production of
a
combined gel for the treatment of infectious diseases of the mucous
membrane of
the mouth and gums. Hazard Analysis and Critical Control Points (HACCP)
were
used to evaluate and manage risks in the manufacture of the new drug.
The HACCP
tool, the “Decision tree”, was used to establish critical control
points. Results
of the research and discussion. An expert
group of experts conducted an analysis and evaluation of the danger of
individual stages of the technological process and determination of the
criticality of the controlled parameters. The group consisted of
qualified
specialists from the pharmaceutical development department, thefactoryworkshop
for the production of soft medicines, the quality control department. Analysis
of the technological scheme of production of combined dental, showed
that almost all stages of the dental gel
manufacturing process are critical and marked in grey. Using the tool
“Decision
tree”, we identified the critical control points of the technological
process
of gel production, and set the eligibility criteria. Risk factors were evaluated on the basis of two
indicators: the likelihood
of a hazard factor and the degree of risk created by that factor. The
likelihood of a risk factor was as follows: unlikely, quitelikely,
probably,
very likely. The degree of risk was assessed on a ten-point scale.
During the
processing of the gel technology, certain critical control points were
monitored
with the aim of developing preventive and corrective actions in case of
their
fall outside the eligibility criteria. in the process of gel
production, risk
(physical factor) at the stages of metronidazole suspension
preparation,
preparation of gel base and gel directly is “Quite likely”. “Unlikely”
occurrence of a dangerous factor at such stages as the weighing of
active and
auxiliary substances, the preparation of solutions and during the input
control
of raw materials. The risk level of each factor, ranging from 2 to 7
points,
was determined and corrective and preventive actions were developed for
all
critical control points to prevent risks to the quality of the
medicinal
product. Conclusions. Based on the
data obtained in the development of dental gel technology and using
scientific
knowledge and methodology of risk assessment by the method of HACCP, an
analysis of technological scheme of its production was carried out.
Process
risks have been identified, critical control points have been
identified and
their allowed limits. For each control parameter, the probability of
occurrence
and the degree of risk were determined, and measures were proposed to
prevent
or eliminate the effects of the risk. Keywords: quality risks, HACCP method,
technological process, control critical points, dental gel.
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24-29 |
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Justification of conditions of
salicylic acid introduction into emulsion ointment base composition
Zuykina E.V., Polovko N. P. Introduction. The revival of extemporaneous production requires the updating of prescriptions, the improvement of the technology of drugs, the study of stability in the process of storage in order to extend the shelf life. Semi-solid dosage forms, which take more than 25 % of medicines prepared in pharmacies about the stock, are currently prepared on a vaseline base. It has many disadvantages compared to the modern emulsion bases that are widely used in European practice. Therefore, the creation and implementation of modern emulsion bases for semi-soled dosage forms is a promising direction for the development of pharmaceutical compouding. To obtain the drug of good quality, it is necessary to provide a rational way of introducing the active substances to the base, so it is necessary to justify the parameters of the ointment with salicylic acid preparation in the conditions of pharmacies. Material & methods The samples of the emulsion bases of the first and second kinds, into the composition of which 20 % of salicylic acid was introsuced by the different technology, were investigated. Determination of the degree of dispersion of the oil phase and salicylic acid in the base was carried out using a “NIKON ECLIPSE CI-S” triocular digital usb microscope with a built-in camera (the lens 40 X / 0.65 160 / 0.17; eyepiece WD 0,56) with 40x magnification. Results & discussion Microscopic imaging of the test samples showed that the average size of crystals in the base of the first kind was 5 times larger than in the base of the second kind. A more uniform distribution of the drug substance in the sample with the base of the second kind was established. It is shown that the more complete dissolution and uniform distribution of salicylic acid in the base are observed when dissolving the drug substance in the emulsion base of the seconf kind. Conclusion. Physico-chemical and microscopic studies substantiated the method of introducing salicylic acid into the emulsion base. The dependence of the degree of dispersion and distribution of salicylic acid on the type of selected emulsion base is shown. It was shown that API is advisable to dissolve in oil, since it promotes less dispersion and a more homogeneous distribution of salicylic acid. The technological scheme of preparation of 20 % salicylic ointment in the conditions of pharmacy is developed, which is layed in the basis of the technological instruction. Key words: technology, semi-solid dosage forms, microscopic examination, emulsion base. |
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30-34 |
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Modern pharmacotherapy of chronic hepatitis
C in patients who failed to achieve sustained virologic response
Kireyev I.V., Zhabotynska N.V. Introduction. According to WHO experts, about 150 million people suffer from chronic viral hepatitis C (CHC), and 350,000 die annually as a result of liver damage by the hepatitis C virus (HCV). Ukraine is one of the countries with medium prevalence of CHC – about 3% of citizens are infected. CHC has become a treatable disease with the use of antiviral drugs (> 95%). To date, for the pharmacotherapy of CHC, a combination of pegylated interferon (PEG-IFN) with ribavirin and direct-acting antivirals (DAAs) are used. Pharmacotherapy of ÑHC using a combination of PEG-IFN and ribavirin has a relatively high efficiency, but it depends on the genotype of the HCV. Therefore, the use of DAAs is a priority in pharmacotherapy of chronic hepatitis C. Patients who failed to achieve sustained virologic response (SVR) are given a second course of treatment (retreatment). The decision on this is based on the following main positions: the nature of the previous response, the type of previous therapy and the potential for a new type of treatment, the severity of liver damage, the genotype of the virus and the presence of other prognostic factors and tolerance to previous therapy. Material & methods. The article analyzes the recommendations of the American Society of Infectious Diseases (IDSA) and the American Association for the Study of Liver Diseases (AASLD), in collaboration with the US International Anti-Virus Society (IAS-USA), as well as the WHO recommendation for repeated pharmacotherapy of CHC in patients who failed to achieve SVR. Results & discussion. To date, for re-treatment of CHC in patients receiving treatment without achieving a SVR, the different re-treatment regimens are recommended depending on the genotype of the HCV. An important problem during pharmacotherapy of patients with CHC is resistance to antiviral therapy. The amino acid polymorphism of NS3, NS5A and NS5B viral proteins in different HCV genotypes and subtypes, as well as the same strains of genotypes and subtypes that reduce DAAs efficacy, is referred to as resistance-related variants (RRV). However, antiviral therapy fails only when RRV is combined with other factors and features of the patient's body, decreased sensitivity to antiviral therapy, or insufficient duration of therapy. As seen in the recommendations for recurrent CHC pharmacotherapy, possible resistance to the protease inhibitor NS5A and to the NS3 protease inhibitors was considered. Conclusion. The results obtained from published sources indicate that current strategies for recurrent pharmacotherapy of CHC patients in most cases of unsuccessful pre-treatment allow the achievement of SVR using DAAs during re-treatment, including those regimens that have efficacy in resistance-associated variants. Key words: chronic viral hepatitis C, re-treatment, sustained virologic response.
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35-38 |
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Photodynamic inactivation
of P. aeruginosa strains in order
to obtain multistage vaccine Derkach S.A., Martinov A.V.,
Gorodnytska N.I., Kutsai N.M., Gabisheva L.S. Introduction. One of the main pathogens of
purulent-inflammatory production, while in the interior remains Pseudomonas
aeruginosa. The pathogen has a serogroup landscape, uses a
polyresistant of
its own and high impurities to disinfectants, asepsis and factors that
exist in
most. When antibiotics are detected, the effectiveness is insufficient,
and
trust in the hymen is created on the early and burn surfaces, on
catheters, in
the formation of a chronic course, which is in overuse of products, and
in the
absurd, pneumonia, implant differences and problems with prosthetics,
which
still have to continue. The problem with creating highly effective
vaccines
against Pseudomonas aeruginosa infection is the availability of
vaccine
antigens that have shown a protective response independent of serotype
and
under the type of pathogen, and have been found to be malignant. toxic
and
non-reactogenic. Look for new products aimed at creating the most
modern drugs
that are studied in different countries. In Ukraine, there are no diagnostic drugs for
the identification of the pathogen and the determination of specific
antibodies,
and vaccines for the prevention and treatment of Pseudomonas
aeruginosa
infection Obtaining such drugs of domestic production is promising,
relevant
and socially and economically justified. Materials and methods.
The
basis of our proposed method of obtaining a multistage Pseudomonas
aeruginosa vaccine is the method of photodynamic disinfection of
bacterial
cultures. An important feature of this method is its universal
effectiveness of
inactivation of bacteria and viruses, regardless of serotype, phagovar.
For the
experiment, 7 regional strains of P.
aeruginosa isolated from different habitats stored in
purulent-inflammatory
sites were studied. As drugs for obtaining samples of the vaccine used
a
commercial drug - "Pseudomonas aeruginosa bacteriophage" (FDUP
"NGO" Microgen ", Perm). As photosensitizers used 1% solution of
vikasol (manufactured in Ukraine, "Darnitsa") and 0.1% solution of
riboflavin (manufactured in Ukraine, "Darnitsa"). To use the
photodynamic effect, we used a photo-polymer stand-alone lamp "Lux"
with a powerful luminous flux of 1200 mW / cm2 (power - 90 mW / cm2.
Ultraviolet (UV) radiation was used in the laminar box with a
bactericidal
lamp.. The titer of a mixture of our adapted phages was determined (by
the
Appelman method), which allowed to determine the dose of phage required
for
testing the phagolysis technique in experiments with various additives
(riboflavin, vikasol). Results and discussion. At a phage titer
of 1: 106,
complete lysis of test-culture pseudomonads was recorded after 2-18
hours of
incubation at a temperature of + 350 C. It should be noted
that the
growth of single colonies still occurs when sowing from a mixture of
pseudomonads + adapted bacteriophages for 3-5 days. . Without irradiation, these substances in any
concentrations did not affect the growth of Pseudomonas aeruginosa
cultures, while at certain doses (vikasol - 3.5 μg / ml, riboflavin -
0.2 μg /
ml) and irradiation parameters there was a decrease or lack of growth
of
bacterial cultures after sowing from experimental samples. The type and
parameters of crop irradiation were determined separately. The
irradiation
regime was determined experimentally on pure cultures of P.
aeruginosa strains
taken in different concentrations (from 101 to 109)
CFU /
ml, at different time intervals (5,0-10, 0-15, 0-20, 0-30, 0 - 40, 0
min), with
different concentrations of riboflavin and vikasol, with the addition
of
bacteriophage samples and without its addition). The optimal parameters
for
irradiation of the samples are 20 minutes when using UV rays and 30
minutes -
when using daylight photopolymer lamp. Significant advantages of one
method
over another are not presented. To increase the photosensitizing effect, the combined use
of viêasol and
riboflavin was used. The most promising way to obtain a phagolytic
pseudomonas
vaccine that is decontaminated and does not contain toxic fractions is
the use
of specific or adapted to candidate cultures P. aeruginosa bacteriophages,
riboflavin and vikasol, followed by irradiation with light or
photopopulation.
The method of obtaining immunogens developed on the model of P.
aeruginosa
can be successful for obtaining vaccines from other bacteria -
pathogens of
purulent-inflammatory diseases (staphylococcus, streptococcus,
Escherichia
coli, Proteus, etc.) and the design of polyvalent vaccines. Keywords: Photodynamic inactivation, Pseudomonas aeruginosa, multistage vaccine
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39-42 |
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Development of a combined diphtheria vaccine based on
diphtheria
anatoxin with microbial adjuvant in the light of modern strategies of
vaccines
development Yelyseyeva I.V., Babich Ye.M., Zhdamarova L.A.,
Bilozersky V.I., Kolpak
S.A. The review article is devoted to the consideration of
approaches to the practical implementation of the main provisions of
modern
ideas about the role and mechanisms of innate immunity for the
development of vaccines. Initiation of the
body's immune defenses
occurs through the recognition of microbial pathogens using typical
molecular
structures associated with pathogens (PAMPs), mediated by image
recognition
receptors (PRRs), which are expressed by cells of the innate immune
system. Òhe
so-called Toll-like receptors (TLR) play a central role in initiating
immune
responses. Other PRRs such as membrane-bound lectin C receptors (CLR),
cytosolic proteins such as the nucleotide-binding oligomerization
domain - NOD-like
receptors (NLR) and RIG-I-like are involved in the recognition of PAMP
and the
control of innate immunity. receptors (RLR), AIM-2-like receptors, and
a family
of enzymes that function as intracellular nucleic acid sensors,
including OAS
and cGAS proteins also involved in the recognition of PAMP
and control of innate immunity. Innate control of adaptive immunity is
now an
established paradigm. PRR determines the origin of antigens recognized
by
receptors expressed on T cells and B cells, as well as determine the
type of
infection with which collides with the body, and teaches lymphocytes to
induce
an appropriate effector class of the immune response. A modern branch of vaccinology is a new paradigm for the
development of
broad-spectrum prophylactic drugs based on trained immunity (TIbV).
These are
vaccines that induce the learning or training of innate immune cells,
the
essence of which lies in their long-term metabolic and epigenetic
changes,
which lead to an enhanced cellular response to the second antigenic
stimulus by
the same or unrelated specific microbial stimulus. Because trained
immunity is
typically triggered by PRRs, TIbV
must be
formed from microbial structures containing the appropriate PRR
ligands,
namely, PAMPs. Unlike conventional vaccines, which aim to obtain only
specific
responses to vaccine-associated antigens, TIbV aims to stimulate a
wider range
of reactions. Broad protection can be achieved by enhancing the
nonspecific
effector response of innate immune cells to pathogens and using the
dendritic
cells activation state to enhance the adaptation of the T cell response
to both
specific and unrelated antigens. The
concept of TIbV
is in its infancy, but a number of modern anti-infective vaccines,
immunomodulators and vaccine adjuvants can already be considered from
the
standpoint of the TIbV category. Induction can be achieved in various
ways that
enhance immunity, which can be involved by bacteria, fungi (β-glucan)
or
metabolic "trainers", as well as some cytokines. A new
paradigm for drug development and therapeutic interventions for the
prevention
and treatment of infectious diseases is also the defeat of bacterial
virulence
as an alternative to antimicrobial therapy. One of the many
antivirulence
targets is adhesion. If it is possible to suppress adhesion,
accordingly, it is
possible to suppress the corresponding infection. This approach forms
the basis
of anti-adhesive strategies that have been invented to prevent various
bacterial infections. Thus, the development of vaccines that prevent
the
initial stage of infection is in line with the anti-adhesive strategy.
A
combined diphtheria vaccine based on native purified diphtheria toxoid
(NODA)
with an adjuvant of microbial origin is developed at the SI “IMI NAMN”
at the
stage of preclinical trials. As an adjuvant in the candidate vaccine, a
preparation of C.diphtheriae, var.gravis, tox + native surface antigens
is
used, which have not undergone modification or even denaturation with
chemicals
and therefore contain molecular structures as similar as possible to
natural
PAMPs and provide targeted antigen stimulation of cells of the innate
immune
system, obtained by physical means of disintegration of microbial cells
(ultrasound or electromagnetic radiation of extremely high frequency). Studies have shown that the nativeness of antigenic
candidate drugs for adjuvants is a key element of their effectiveness.
Experimental samples of diphtheria bacterial surface antigens have
shown
themselves as adjuvant for diphtheria toxoid, capable of replacing
neurotoxic
aluminum hydroxide, and immunomodulator, which increase phagocytic
activity in
the first and repeated antigenic stimuli, and also promote the release
of
nasopharyngeal mucosa of rabbits vaccinated with experimental samples
of the
combined diphtheria vaccine and infected with a culture of C.
diphtheriae. The
formation of an immunologically strong mucosal barrier is considered to
be an
effective strategy to prevent infection at the point of contact between
microbes and the host. However, modern standards of vaccine technology
usually
apply only to pathogens that have already crossed the mucosal barrier.
In
contrast to licensed vaccines, vaccination on mucosal surfaces,
including the
oral route of administration of the vaccine, can successfully stimulate
the
humoral and cellular immune response in both systemic and mucosal areas
of the
entrance gate of infection to establish a broader and longer-lasting
protection. Experimental samples of the
combined
diphtheria vaccine with bacterial adjuvant were tested using the oral
route of
administration of the vaccine in combined vaccination regimens, and, as
the
results of experiments showed, the adjuvant and phagocytosis
stimulating effect
of the studied candidate vaccines was maintained. The obtained data
demonstrated
the prospect of widespread use of oral immunization as the most
natural,
physically and psychologically painless way to administer vaccines both
for
emergencies in the diphtheria outbreak and to maintain collective
diphtheria
immunity by booster immunization. However, oral delivery is a
complex
task that requires a special composition to overcome harsh
gastrointestinal
environments and avoid the induction of tolerance to achieve effective
protection, which requires detailed justification of oral vaccines,
including
key biological and physicochemical aspects of next-generation oral
vaccines. Another way to increase the safety of diphtheria
vaccines during repeated vaccinations, which opens up prospects for
specific
immune protection of persons with allergic reactions, was the
experimental
application of the principles of specific immunotherapy in vaccinations
of
experimental animals. The obtained results indicate that the previous
oral
administration of antigenic drugs C. diphtheriae prevented the
development of
allergic skin reactions in experimental rabbits with subsequent
subcutaneous
administration of diphtheria vaccines. Studies on the development of a
combined
diphtheria vaccine with an adjuvant of bacterial origin convincingly
showed the
immunogenicity of the obtained experimental samples of the candidate
vaccine
and the possibility of combining the efficacy and safety of the drug at
a
certain dosage and degree of purification of antigenic drug. But for
further
research, it will be essential to expand knowledge about the sensitive
pathways
of the innate immune system and to determine the rules of interaction
that
determine the functions of these pathways in the context of diphtheria
infection. Special attention should be paid to the study of the
protective
effect of the drug, the study of patterns of formation of antibacterial
diphtheria immunity and the determination of optimal ratios of NODA and
bacterial antigen in the vaccine. Key words: trained innate immunity, adaptive immunity, development of vaccines, bacterial adjuvants, C.diphtheriae, mucosal vaccines, specific immunotherapy.
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43-49 |   |
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The stay of
humoral immunity in bacterial dysbiosis and bacterial vaginosis Hruzevskyi O.A., Minukhin V.V. Introduction. The state of dysbiosis and bacterial vaginosis (BV) is
characterized
by the formation of both systemic and local immune deficiency, which
corresponds to the increase in the number of pathogenic microbiota. An
important
reason of bacterial vaginosis’ development is local immunodeficiency
corresponding
with decreasing of colonization resistance of vaginal fluid. This phenomenon develops due to disturbance of
normal vaginal microbiocenosis, secretion of antimicrobial substances
and provision
of normal immune defense. Recognition of the role and mechanisms of
local immunodeficiency’s
development can be very important scientific achievement in the field
of
microbiology, immunology and pathology of human vaginal microflora.
However,
nowadays ratio of systemic and local immune reactions in bacterial
vaginosis isn’t
revealed completely. Thus, the aim of
the investigation was to determine the stay of humoral immunity
according to
the content of immunoglobulins (Ig) in the blood and vaginal fluid in
different
degrees of bacterial dysbiosis and BV. Material
and methods. Data from 298 women were divided into groups according
to
index of pathogenic microbiota condition (IPMC) and the pathogenic
microbiota
indicator (PMI): normocenosis (n=53), dysbiosis I (n=128) and II degree
(n=117), among the last allocated 83 patients with PMI>1
lg gE/sample,
where was drawn diagnosis “Bacterial Vaginosis”. The criterion of exclusion there was presence of
pathogenic
microorganisms in vaginal epithelium scrapings. These representatives
were: Trichomonas vaginalis,
Neisseria gonorrhoeae, Chlamydia trachomatis òà Herpes Simplex Virus 1,2.
Presence of leucocytes more than 15-20 cells in the field of vision in
vaginal
smears indicated inflammatory reaction also was the criterion of exclusion. Molecular genetic studies of posterolateral wall of the
vagina epithelium
scrapings was performed by Real-time polymerase chain reaction. A
content of
facultative and obligate anaerobic bacteria, myco- and ureaplasma,
yeast-like
fungi was studied quantitatively. With the help of Enzyme-Linked
Immunoassay
(ELISA) contents of IgA, IgM,
IgG, IgG2, secretory IgA (sIgA) were determined in blood and
vaginal
fluid. Spectrophotometry was used for quantitative evaluation of
circulating
immune complexes (CIC) contents in the blood and immune complexes in
vaginal
fluid (ICVF). For descriptive statistics of
data there were used arithmetical mean (Ì) and average error
(mistake).
Paired independent data
samplings were compared according to Mann-Whitney U-test
(U). Significance of all differences
accepted when p<0,05. For statistic and regressive analyses package
of software
“Statistica 10” (StatSoft, Inc., USA) was applied. Results
and discussion. While development of bacterial dysbiosis and
BV there was observed progressive increasing of CD22 lymphocytosis, contents of IgM, IgG ³ IgG2.
In our investigations quantity of CD22+ lymphocytes was constantly larger in manifested dysbiosis in
comparison with normocenosis.
Maximal content of CD22+ lymphocytes was noted in BV. Contents of IgA, sIgA and CIC had tendency to decreasing. In
general, it’s possible
to conclude that blood CIC level decreases according to the progressing
of
dysbiosis. Hence, its more level in 2 subgroup of 2 group and in 1
subgroup of
3 could indicate reactive changes in immune system. Content of ICVF
in
I degree dysbiosis in comparison with normocenosis was not change
significally.
Simultaneously, in I degree dysbiosis and in 1-st subgroup of II degree
dysbiosis
this inex was significally more. In BV content of ICVF was
twice less
than in normocenosis. These phenomena were synchronous with blood CIC
levels, and
reflected sharp parallel decreasing of CIC formation both in the
bloodstream and
in vaginal fluid during BV. Local immunodeficiency with
immunoglobulins’ (especially,
IgA and sIgA) and ICVF levels decreasing progressed while development of bacterial dysbiosis and
BV also.
Therefore, the stay of systemic humoral immunity in BV didn’t correlate
always
with such one in vaginal fluid. Conclusion. Systemic humoral immunity while development
of BV was changed, but it wasn’t reflected completely the stay of
defences in
vaginal fluid. In general, there was present dissonance of these two
systems’ reaction:
activation of systemic level and suppression on local level. Key words: bacterial vaginosis, humoral immunodeficiency,
vaginal dysbiosis |
50-56 | |
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|
Genetic monitoring of endemic
measles virus circulation in European countries Kalinichenko S.V.,
Melentyeva K.V., Toryanik I.I., Zvereva N.V., Antusheva T.I., Buriachenko
S.V. Introduction. The measles
virus is still one of the main causes of morbidity and mortality in
children
and adults and is a threat of infectious outbreaks in many countries
around the
world. The World Health Organization (WHO), at the 2015 meeting in
Europe, set
out to eliminate measles infection. To control the elimination of this
disease
requires the accumulation of genotyping data of the detected measles
virus to
interrupt the situation of endemic spread. All six WHO regions have set
a
target for combating measles. In order to monitor and evaluate the
degree of
endemic circulation of measles virus (MV), the transmission chains of
the
epidemiologically relevant variants of MV identified in Central and
Western
Europe are analyzed. More systematic molecular monitoring and recording
of MV
transmission data between many countries can help to create a
meaningful
picture of the process of eliminating the problem of the occurrence and
spread
of measles infection. Goal. To study
whether molecular surveillance meets the challenge of eliminating
measles
infection with the assurance of molecular data quality, continuity and
intensity of molecular monitoring and analysis of transmission chains
in
different geographical regions. Material
& methods. Published articles, molecular
program for external WHO
quality assessment, WHO EUR central infectious disease information
system, and
WHO measles surveillance database. Results & discussion. According to
the WHO standardized nomenclature using the nucleotide (nt) sequences
of the N
and H variable genes, wild-type measles viruses are currently divided
into 24
genotypes. The most variable is the 450-nt variable coding sequence of
the
C-terminal portion of the N protein (N-450 region) and is used to
differentiate
detected MV for observation. Antigenic differences between measles
virus
strains - representatives of different genotypes are minimal, all known
genotypes of the virus belong to one serotype. Since the beginning of
molecular
surveillance in Europe in the early 1990s, only two genotypes of MV (C2
and D6)
have been identified, which have been spread throughout the region and
are
therefore called indigenous European genotypes. Molecular
observation has shown that, over the years, the endemic genotypes C2
(IR /
Kempten.DEU / 23.00) and D6 (IR / Berlin.DEU / 47.00) have changed
rapidly with
the circulating D7 genotype (IR / Mainz.DEU / 06.00) to the beginning
of 2003. The
imported measles virus of genotypes B3, D4, D5, D6, D8, D9, H1 appeared
in
Germany from 2005 to 2009 - 2010. Most cases were related to the
measles virus
of genotype D4, and its several sub-variants. According to the
monitoring data,
genotypes D8, B3, H1, D9, D4 have been circulating in the world in
recent years
(from August 2017 to July 2018). Of
the 179 measles deaths reported in European countries during 2009-2018,
114
(64%) occurred during 2017-2018, including 93 (82%) in four countries:
Romania
(46), Ukraine ( 20), Serbia (15) and Italy (12). EU countries report
17587
measles virus sequences to the WHO global measles surveillance
database. The
most common measles virus genotypes were D4 (21% overall, 66% in
2009-2012), D8
(45% overall, 76% in 2013-2016) and B3 (33% overall, 58% in 2017-
2018). Conclusion. Our research illustrates
the long-term transmission of MV in Europe. Which probably happens
because of
the unvaccinated people in the various hard-to-reach groups that
transmit the
infection to the general population. This situation is, of course,
inconsistent
with the purpose of the WHO and UNESCO (WHO-UNICEF, 2002) measles
elimination
program already achieved in America and Australia. In order to address
the
global problem of measles infection worldwide, additional efforts are
needed to
identify deficiencies in immunization among the population. As the
elimination
of MV should be a problem for all EUR countries, similar research to
ours
should be expanded to obtain comprehensive information on the
circulation of MV
strains in different regions across Europe. Keywords: Genetic monitoring, measles, circulation, Europe
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57-70 | |
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The effect of Bifidobacterim bifidum
cell-free supernatants, ascorbic acid,
fructose, sorbitol, xylitol and stevia on the daily biomass growth of opportunistic microorganisms Knysh O.V., Martynov A.V. Introduction.
The
use of probiotic bacteria as biotransformator
system is a promising way to obtain
new derivatives of known substances with antimicrobial or other
beneficial
activity.
The aim of
the research was to investigate the effect of the Bifidobacterim
bifidum cell-free supernatants, obtained by cultivation of
bifidobacteria
in their own disintegrate supplemented with ascorbic acid
(BbAsc), fructose (BbFr), sorbitol (BbSor),
xylitol (BbXyl) or stevia (BbSt) and the substances themselves (Asc,
Sor, Xyl
or St) on the growth of opportunistic microorganisms.
Material
& methods. The effect
of the studied supernatants and substances on the daily biomass growth
of the
test cultures was investigated by spectrophotometry using a 96-well
polystyrene
microtiter plates and a «LisaScanEM» spectrophotometer
(«ErbaLachemas.r.o.»,
Czech Republic). Reference strains Staphylococcus
aureus ATCC 25923; Escherichia coli AÒÑÑ
25922 and Pseudomonas
aeruginosa AÒÑÑ 27853 were
used as a test cultures. The final concentration of the studied
supernatants
and substances in the incubation medium was 30%vol, and the final
concentration
of bacterial cells was ~106 CFU/ml. Results
& discussion.
Among the studied
substances, stevia alone did not affect the
daily biomass growth of any of the test cultures, either as in solution
with a
final concentration of 30 mg/ml or as in a supernatant of B. bifidum
culture. Fructose at a final
concentration of 30
mg/ml had the same effect on test cultures growth as the supernatant of B.
bifidum culture after probiotic cultivation with fructose: it
inhibited staphylococcal
growth (Fr: II = 30,9 %, BbFr: II = 34,6 %),
stimulated the growth of P. aeruginosa
(Fr: ²S = 23,7 %, BbFr: IS = 19,8 %) and
did not affect the growth of E. coli. Xylitol at a final
concentration of 30 mg/ml did not affect the
growth of P. aeruginosa biomass, but
inhibited the growth of E. coli (²I = 20,9 %) and S. aureus (²I = 28,4 %). The supernatant of B. bifidum, cultured
in the presence of
xylitol inhibited the growth of S. aureus
biomass (²I = 26,1 %) and
did not affect the growth of the other test cultures. The sorbitol and cell-free
supernatant of B. bifidum cultured in
the presence of this polyol equally influenced the biomass growth of
test
cultures: had no significant effect on the daily biomass growth of S. aureus but stimulated the growth of E.
coli (BbSor: ²S = 43,9 %, Sor: ²S = 34,5 %)
and
P. aeruginosa (BbSor: ²S = 26,4 %, Sor: ²S = 28 %). The only substance that
significantly inhibited the biomass growth of all test cultures was
ascorbic
acid. Cell-free supernatant of B. bifidum (BbAsc) cultured in disintegrate
supplemented with ascorbic acid caused more pronounced inhibition of
the daily biomass
growth (S. aureus – 78 %, E. coli – 52
% and P. aeruginosa – 45 %) compared to ascorbic acid
(Asc) itself (S. aureus – 42 %, E. coli
– 35 % and P.
aeruginosa –
38 %). Conclusion. Among
the studied substances, only ascorbic acid caused significant
inhibition of
biomass growth of all test cultures. B.
bifidum sell-free supernatant obtained by cultivation of
bifidobacteria in their own disintegrate
supplemented with ascorbic acid caused a more severe inhibition of test
cultures
growth than ascorbic acid itself. The
absence of signs of chemical modification of ascorbic acid in the B. bifidum cell-free supernatant on the
chromatogram indicates that the greater inhibitory effect of this
supernatant
is due to the synergistic effect of ascorbic acid and the inhibitory
compounds
produced by B. bifidum during
cultivation. The other studied substances showed various effects on the
daily
biomass growth of test cultures. The use of B.
bifidum as a biotransformer system, and xylitol, sorbitol, fructose
and
stevia as precursors for the production of new antimicrobials by
combinatorial
biosynthesis, turned out to be not effective enough. Keywords: Bifidobacterim bifidum, cell-free supernatants, ascorbic acid, fructose, sorbitol, xylitol stevia, biomass growth, opportunistic microorganisms
|
71-76 | |
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|
Clinical
and microbiological studies of aminoglycosides efficiency in purulent
inflammatory processes Dmytriiev D.V., Nazarchuk
O.A., Babina Y.M, Maistruk S.B. Introduction. Infectious complications of
diabetic foot
syndrome are one of the main complications of diabetes mellitus, as
well as a
significant risk factor for amputation of the lower limb. The use of
effective
antimicrobial therapy is an important component in the treatment of
these
infections. Knowing the problem of
antibiotic resistance in our time, the choice of starting antimicrobial
drug is
very important to reduce ineffective treatment, resistance to
antibacterial
agents, unwanted complications and economic costs. Objective. Conduct a comparative microbiological study of
the antimicrobial efficacy of aminoglycosides against pathogens of
wound
suppurative-inflammatory processes and determine the clinical
effectiveness of
these agents. Materials and methods. The
study included 45 patients with
purulent-inflammatory processes of wounds in diabetic foot syndrome who
received surgical treatment and antibiotic therapy.
Patients were randomly assigned to three
groups according to the prescription of antibiotics from the class of
aminoglycosides
(tobramycin, amikacin, gentamicin).
Clinically took into account the general condition of the
patients,
wound healing, laboratory parameters (blood test, procalcitonin, CRP). Microbiological research was carried out in
the bacteriological laboratory of the Department of Microbiology. Microbiological identification of the
isolated microorganisms was carried out by the classical method
according to
morphological, tinctorial, cultural, biochemical features. Results
& discussion.
Microbiologically established polymicrobiality of wound contents in
patients
with diabetic foot Syndrome.
Gram-positive and gram-negative microorganisms were
identified. Aminoglycosides (gentamicin,
tobramycin,
amikacin) provide the same bacteriostatic, bactericidal effect on
sensitive
clinical strains of S. aureus, S. epidermidis, E. coli, E. cloacae, P.
aeruginosa, K. teriggena, tobramycin had the advantage of antimicrobial
activity to A baumannii, E. faecalis,
and resistant strains of staphylococcus (p <0.05).
As a result of studies, good tolerance by
patients to antibiotics from the aminoglycoside group as starting
monotherapy
in 91.8% was revealed. according to A.
baumannii, E. faecalis, and resistant staphylococcus strains (p
<0.05). Clinically, tobramycin also
showed advantages
in local wound healing and, according to laboratory data (a decrease in
the
level of leukocytosis, procalcitonin and CRP), is 30% more effective in
the
first 6 days of antibiotic therapy, compared with group 2 and 3 of the
study. Conclusion. Considering the
polymicrobial spectrum of
infectious pathogens in diabetic foot syndrome and the antibiotic
resistance of
the use of aminoglycosides in monotherapy for mild to moderate
processes and in
combination therapy with other antibiotics for severe degrees of
complications,
this is a balanced strategy for preventing the development of microbial
resistance. Aminoglycosides (gentamicin,
tobramycin, amikacin) provide both bacteriostatic and bactericidal
effects on gram-positive
and gram-negative pathogens, and especially tobramycin, which has the
advantages of antimicrobial activity against A. baumannii, E. faecalis,
and
resistant strains of staphylococcus. Keywords: diabetic foot syndrome, gram-positive pathogens, gram-negative pathogens, antibiotics, aminoglycosides.
|
77-85 | |
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|
Bioinformation analysis of polymorphism of the blaTEM gene coding regions in Escherichia
coli strains Peretyatko O.G., Yagnuk
Y.A., Sklyar N.I., Bolshakova G.M., Cholodna T.V. Bioinformation
analysis of the bacterial genome with screening of genetic elements to
detect
variability of nucleotide sequences is one of the methods of antibiotic
resistance
evolution fundamental research. Comparative analysis of the molecular
genetic
variability or genome sequence conservatism is used as a convenient
tool for
understanding patterns of the evolutionary process through definition
of the phylogenetic
relationships. The aim of the study
was to conduct a bioinformation analysis of the variability of the
antibiotic
resistance blaTEM gene in E.
coli strains. Materials and methods.
The GenBank nucleic acid search database was used for bioinformation
analysis.
The search for nucleotide sequences of the blaTEM
gene was performed in fasta-format. "Vector NTI Advance 11.0"
software package was used to analyze the nucleotide sequences of the blaTEM gene. Multiple alignment and its
statistical analysis to determine conservative and variable regions of
the blaTEM gene was performed using software
component module “AlignX”. Nucleotide sequences of the 21 blaTEM
gene sequences isolated from E. coli strains in 11
countries and registered in the GenBank
database from 1996 to 2019 were analyzed. Research
results and discussion. Multiple alignment of the nucleotide
sequences of
the studied blaTEM gene sequences
allowed to determine both conservative and variable regions of the
gene. 12
conservative regions of the blaTEM gene
with a length of at least 30 nucleotide sequences were identified. The
longest
homologous region of the gene (144 nucleotide sequences) was found at
the 529 -
672 nn position. blaTEM gene
variability was evidenced by point mutations at 41 positions of the
nucleotide
sequences. The most heterogeneous region was found at the 697-717 nn
and
773-813 nn positions. Mutations in the analyzed E. coli
blaTEM genes were
caused by transitions (60,5%), transversions (37,2%) and deletions
(2,3%). The
degree of relatedness of the analyzed blaTEM
gene sequences is presented in the form of a dendrogram. The
phylogenetic
tree shows three conditional genetic clusters. The top position of the
dendrogram is occupied by the cluster that includes eight blaTEM
gene sequences, isolated in European countries: 2 - in
France and one each - in Poland, Germany, England, Portugal, Italy and
the
Netherlands. This cluster was characterized by a significant number of
evolutionary events (from 4 to 9) associated with nucleotide sequence
substitutions,
which is represented on the dendrogram by the divergence of the
phylogenetic
tree branches. The cluster which occupies the middle position of the
dendrogram
is represented by five phylogenetic branches. This cluster includes blaTEM gene sequences isolated in Korea,
China, Germany and India. 1-2 nucleotide substitutions were identified
in the
majority of the sequences in this cluster. The lower position of the
dendrogram
consists of blaTEM gene sequences
characterized by a high degree of relatedness and a small number of
nucleotide
sequence substitutions, a significant number of sequences were isolated
in the
United States and China (75%). Conclusions.
Results of the bioinformation analysis showed that the
nucleotide sequences
of the E. coli blaTEM gene have
historically undergone several evolutionary divergences and acquired
signs of
heterogeneity, which apparently causes the emergence of a large number
of
species of the blaTEM gene. Key words: bioinformation analysis, E. coli, blaTEM gene.
|
86-89 | |
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|
Development of a method for determining the probability
of a decrease in
post-vaccination immunity in individuals with herpesvirus load Smilianska M. V., Kashpur N.V., Peremot S.D., Khodak L.A., Naviet T.I. Introduction. The most effective way to combat infectious diseases is
to vaccinate the
population. Each country develops its own vaccination schedule, which
takes
into account the specifics of the epidemic situation, the availability
of
registered vaccines, financial opportunities and other factors. Experimental and
clinical studies have shown that vaccines can cause both suppression
and
activation of individual immune functions, with each vaccine having its
own
spectrum of effects on the quantitative and functional characteristics
of the
immune status. There is a need to correct the formation of immunity
during
vaccination, taking into account factors affecting the intensity of a
specific
response to vaccine administration. It is proposed to use the
principles of
individualization of vaccination (revaccination), primarily in
high-risk
groups, which include people with persistent herpes virus infection. Material & methods. An immunological
examination was carried out, including determination of the
concentration of
cytokines TNFα, IL10, IFNγ, levels of CD3 + CD4 + and CD3 + CD8 +,
levels of
subclasses 1 and 3 of specific IgG antibodies, viral load (HVL). The
study
material was blood in a volume of 3.0-5.0 ml, which was taken from a
vein in
compliance with the usual rules of asepsis. To determine the concentration of TNFα, IL10, IFNγ
cytokines in blood serum, the Vector-Best ELISA test systems were used:
Gamma-interferon IFA-BEST, interleukin-10 - IFA-BEST, Alpha-TNF
IFA-BEST.
Herpesviridae family antigens (Ag) were determined by
immunofluorescence using
specific monoclonal mouse antibodies from Santa Cruz Biotechnologu,
Inc. (USA)
and viral load (herpes viral load, HVL). CD3 + CD4 + and CD3 + CD8 +
were
studied by flow cytometry on a CYTOMICSFC500 (Beckman Coulter, USA)
using the
MKAT panel (Beckman Coulter, USA). Subclasses of specific IgG
antibodies are
determined by ELISA in modification. In this case, 96-well panels
coated with
antigens are used, respectively, according to a commercial kit for the
determination of specific IgG antibodies (Euroimmun or Human, Germany).
Serum
is added at a 1:50 dilution. The conjugate is peroxidase-labeled
anti-IgG1,
IgG2, IgG3 and IgG4 monoclonal antibodies (“Polygnost”) at a
concentration of 1
μg / ml [9]. Calculation formulas for determining the dynamics of the
complex
integral indicator over time were obtained from the initial data for
the
determination of the levels of specific vaccination antibodies and
complex
integral indicator using mathematical modeling of linear and non-linear
functions of the exponential distribution. Results & discussion. According to the results of studies, we propose to
determine the ratio of such immunological parameters: specific IgG1sp /
IgG3sp;
immunoregulatory index CD4 / CD8; TNFα / IL10, IFNγ / IL10 ratio and
herpes
virus load HVL (herpes viral load); Using the formula IgG1sp / IgG3sp *
CD4 /
CD8 * 0.1 * TNFα / IL10 * 0.01 * IFNγ / IL10 * HVL (c.u.), the complex
integral
index (CII) is calculated. The probability of a decrease in the
post-vaccination immunity of the subject is judged by the change in the
complex
integral indicator over time (according to the chart). Conclusion. The development of
a new method for determining the probability of a decrease in
post-vaccination
immunity in individuals with herpesvirus load is aimed at increasing
the
assessment of the body's immunity against controlled infections and
helps to
decide on the need for revaccination or its delay for a certain period
of time,
which is calculated individually. The end result is a reduction in the
incidence of infectious diseases. Keywords: herpes virus, postvaccination effects, immunity
|
90-93 | |
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|
Intravascular inflammation
- from Mechnikov's idea to the present day Peremot S. D., Smilianska M.
V., Kashpur N. V., For more than a century, the name of I.I.
Mechnikov has been shining in the constellation of the names of
outstanding
scientists, whose works laid the foundations for the formation of a
scientific
worldview of all living things: from a small microorganism to a person.
He is
considered the founder of comparative pathology and the theory of
immunity, a
scientist who stood at the origins of evolutionary embryology,
microbiology and
immunology. He formulated the general theory of inflammation as a
protective
reaction of the body in the fight against infection. The enormous
significance
of Mechnikov’s scientific heritage is based primarily on deep
materialistic and
evolutionary principles, a consistent conductor and passionate champion
of
which he has been throughout his life. Biology and medicine owe the
scientist
not only many brilliant discoveries and firmly facts, but also
significant
broad generalizations laid the foundation for a number of the most
progressive
areas of modern biology and medicine. These include the concept of
«intravascular inflammation». Using this definition, Mechnikov argued
about the
possibility of phagocytosis of bacteria by circulating leukocytes. The
main
idea formulated by him is consonant with the modern concept that,
according to
I. I. Mechnikov, «harmful factors», getting into the blood, excite the
reaction
of phagocytes, the main effector mechanism of inflammation. What
happens with
the classic inflammatory response is «extravasal», thanks to the
emigration of
cells from the bloodstream. He speaks of «intravascular inflammation»
only
once, without focusing attention, however, he testifies to the
correspondence
with the main idea. The scientific thought formulated by Mechnikov,
rethought
and reinforced by the results of studies of his followers, found its
continuation in the modern concept of intravascular inflammation. In
general,
it can be argued that Mechnikov made a scientific feat, the scale of
which
becomes all the more obvious the farther he is from us in the
historical
interval and there is no doubt that the ideas and methodology of our
great
countryman will be key in the 21st century for new fundamental
discoveries in
the field of natural sciences. Keywords: Intravascular inflammation, I. I.Mechnikov, history, immunity
|
94-96 | |
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| Role Ilya Ilyich Mechnikov in tuberculosis
fighting:
Astrakhan results of
the expedition 1911 Peretyatko AG, Yagnyuk YA, Sklyar NI, Bolshakova MA, Kholodna TV Our compatriot, well-known
scientist, Nobel Prize winner in medicine and physiology Ilya Ilyich
Mechnikov
left a bright mark not only in domestic but also in world science. Ilya
Ilyich's multifaceted scientific activity was devoted to various areas
of
research in biology and medicine. The scientist's success in studying
important
problems of zoology, embryology, comparative pathology, gerontology,
immunology, virology and bacteriology is renowned around the world. One of the areas of his research activity aimed at studying
tuberculosis. In
May 1911, the Pasteur Institute organized the Astrakhan expedition led
by I.I. Mechnikov. Leading scientists were among the participants of
the expedition –
specialists in microbiology and epidemiology from Russia, France, Italy
and
Japan. The expedition was to address important issues related to the
spread of
plague and tuberculosis in the Kalmyk steppes. Ilya Ilyich Mechnikov
set the
task to use diagnostic tests (Pirke test and ophthalmological test) to
determine the level of tuberculosis infection in the steppe population
and to
investigate the relationship between the incidence of tuberculosis and
increased contact of Kalmyks with non-steppe population. The work of
the
Astrakhan expedition led by I.I. Mechnikov
became the starting point for successful studies of tuberculosis
infection. When
beginning studying
tuberculosis, I.I. Mechnikov
planned to completely defeat this disease. Although he did not achieve this goal, scientific
conclusions made by Mechnikov while
studying biological properties of the pathogen and mechanisms of immunity to tuberculosis were further
developed, and joint efforts of domestic and foreign scientists made it
possible to elaborate tailor-made measures for particular regions to prevent tuberculosis and justify
the use of antimicrobials
in the treatment of this infection.
Keywords: ².². Mechnikov,
Astrakhan expedition, tuberculosis. |
97-98 | |