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Editorial Board |
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Contents |
2-5 |
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Review |
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Riboflavin in
photodynamic inactivation of pathogens and photodynamic therapy Pogorelaya M, Martynov
A, Romanova E., Mechnikov institute of microbiology and immunology Riboflavin,
besides its key role in providing cellular metabolism
in humans and animals, is used as a
compound of antimicrobial photodynamic therapy (aPDT) owing to its photosensitizing capability. PDT is a promising approach for the photoinactivation of
pathogens in blood
and blood derivatives. It has been reported, that the advantage of
antimicrobial photodynamic therapy is that there are no resistant
strains
to it. Flavins are
photoreducible and photon-induced excitation of them in the
ultraviolet-blue
(UV-blue) spectral band can lead to formation of either singlet oxygen
via
energy transfer to environmental oxygen, or hydrogen peroxide and
derivatives
via radicalisation – altogether termed reactive oxygen species (ROS)
and used
hereafter. Exactly the ROS production property of riboflavin is used
for
antiviral and antibacterial disinfection; for strengthening the corneal
tissue
in photorefractive surgery by the ROS-induced collagen cross-linking.
Several
studies reveal the antimicrobial photodynamic potential
of riboflavin irradiated with the ultraviolet-blue and with visible light against methicillin resistant Staphylococcus aureus,
Staphylococcus
epidermidis, enterohemorrhagic Escherichia
coli, Pseudomonas aeruginosa, Acinetobacter
baumannii and Bacillus subtilis in vitro. It was shown, that riboflavin/UV-A and allowed
effectiveness against such fungal pathogens as Candida
albicans, Candida
parapsilosis, Fusarium spр, and A.
fumigatusas which may cause infection keratomycosis. The photoilluminated riboflavin significantly
reduced the activity of superoxide dismutase (SOD) and reduced the
level of
cellular antioxidant metabolite - glutathione (GSH). Along with that
the
specific activity of glutathione S-transferase (GST) - which is
involved in
detoxification process - was increased significantly in cells exposed
to
photoilluminated riboflavin. Riboflavin,
illuminated by UVA or visible light, has also been developed as a
nucleic acid-binding agent to be used for photoinactivation of such
nucleic
acid-containing pathogens in plasma, platelets, and RBCs as viruses.
Several
studies have revealed the effectiveness of reduction in some viruses’
infectivity, including human immunodeficiency virus 1 (HIV-1), bovine
viral
diarrhea virus (BVDV), hepatitis B virus (HCV), pseudorabies virus. Thus,
the «riboflavin+UVB» system has
found their application in pathogen reduction technology – «Mirasol»®
Pathogen
Reduction Technology (PRT) system. Riboflavin has been thought to be a
promising antitumoral agent in photodynamic therapy, though the further
application of the method was limited by the unclear molecular
mechanism. Several
studies reveal that of PDT-mediated
cytotoxicity occur in three ways:
apoptotic, necrotic and autophagy-associated cell death. Some
findings, show that certain PDT techniques acting via inducing of
apoptotic
cell death that is highly immunogenic and can stimulate antitumor
immunity. Thus,
we can conclude that, the
«riboflavin+UVB» system is suitable for photoinactivation
of bacteria, fungi and viruses and has a potential in
antitumor treatment strategies. Further studies will reveal more and
more
aspects of riboflavin capabilities. Keywords: riboflavin, photodynamic,
antimicrobial, antiviral, UVA, UVB
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6-11 |
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Experimental works |
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Development of technology of plant species for complex
mastopathy
therapy Keywords: technology, medicinal plant species, mastopathy.
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12-17 |
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Quantitative determination of
microbiom in the destination content the gut in rats Bukina Yu.V. Introduction. The gut
microbiome significantly affects the functioning of the body: it
participates
in the protection of the body against pathogenic microorganisms, in the
processes of metabolism, inhibition of inflammatory responses, in the
formation
of innate and adaptive immune response in the intestinal mucosa. One of
the
reasons for changing the microbiota is the use of antibiotics.
Therefore, the
processes of interaction of antibiotics, Salmonella
enteritidis and Salmonella
typhimurium with representatives of normal intestinal microflora
are of
particular interest. Materials and methods. The quantitative
and
qualitative composition of the wall microbiota in rats by
bacteriological
method, the statistical analysis of data using the program StatSoft
Statistica
v12 were conducted. Results & discussion. With the
introduction of
vancomycin and S. enteritidis, S.
typhimurium in groups II, III, IV there was a decrease in E. coli quantitative content by 10, 7
and 110 times, respectively (p≤0.05). The number of P.
aeruginosa decreased significantly only in the third group
(p≤0.05). The number of representatives of Bacteroides
spp. significantly decreased by several thousand times (group II)
and by 70
and 87 times (groups III and IV) (p≤0.05). The content of E.
faecalis and E. faecium
decreased by 861,6 and several thousand times (groups II, III, IV)
(p≤0.05).
The number of Proteus spp.
significantly decreased in group II
by 27 times and increased rapidly in group IV (p≤0.05). Group III
showed a
sharp decrease in the content of representatives of Enterobacter
spp. and Klebsiella
spp. in 847 and 150 times, and in group II there is an increase in
their
number by 7 and 46 times, respectively (p≤0.05). The number of Staphylococcus spp. decreased by 9.8
times only in II group. The quantitative content of Clostridium
spp. decreased by several thousand times (group II) and
by 5.5 times (group IV) (p≤0.05). The number of Lactobacillus
spp. decreased by several thousand times (group II).
The number of representatives of Bifidobacterium
spp. significantly decreased by 10.9 times and by several thousand
times
(groups III, IV). The quantitative content of Peptostreptococcus
anaerobius decreased significantly in all three
study groups (p≤0.05). The content of Salmonella
spp. increased in group II by 49 times and significant increase was
observed in groups III and IV (p≤0.05). The introduction of salmonella,
against
the background of vancomycin pre-treatment, causes a dramatic change in
the
composition of the microbiota in groups V and VI, namely: an increase
in the
number of E. coli 65 and 105 times, a
significant increase in the content of P.
aeruginosa in the V group, and in the VI, 3 times. Also, in these groups there is a decrease in the number of Bacteroides spp. 9 and 10 times
(p≤0.05). The content of E. faecalis
and E. faecium decreased significantly
only in the fifth group (p≤0.05). The number of Proteus
spp. decreases 17 times in group V and also a significant
decrease was observed in group VI (p≤0.05). A sharp increase in the
content of
representatives of Enterobacter spp.
and Klebsiella spp. was observed in
the V and VI groups (p≤0.05). However, representatives of Peptostreptococcus
anaerobius in V and VI groups decreased 20 and 9
times, respectively (p≤0.05). The number of Salmonella
spp. decreased only in group V 7 times (p≤0,05). With the
introduction of
experimental animals B. fragilis treated with S.
enteritidis, S. typhimurium on the background of vancomycin
pre-treatment, a significant decrease in the level of E.
coli in group VII, and in VIII - by 538 times (p≤0.05). The
number of P. aeruginosa in groups VII
and VIII decreased significantly and the number of representatives of Bacteroides spp. naturally increases
(p≤0.05). The content of Lactobacillus spp. decrease
by 10.3
times only in VI group. The content of E.
faecalis and E. faecium increased
by 10 and 19 times in the seventh and eighth groups respectively, and
the
number of Proteus spp. decreases only
in group VII 322 times (p ≤0.05). Also, in VII and VIII groups there is
a sharp
decrease in the content of representatives of Enterobacter
spp. and Klebsiella
spp. (p≤0.05). The level of representatives of Peptostreptococcus
anaerobius and Lactobacillus spp. increased
significantly 7, 12 times and several
thousand and 40 times (groups VII and VIII, respectively) (p≤0.05). The
number
of S. enteritidis and S. typhimurium
in the VII and VIII
groups decreased intensively (p≤0.05). Conclusions. The
introduction of B. fragilis can be used in the
treatment
of inflammatory bowel diseases or diseases with impaired barrier
function of
the intestine. Keywords: parietal microflora, microbiome, vancomycin, salmonella, bacteroids.
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18-26 |
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The influence of polyols on the bacteriotropic
properties of the Lactobacillus reuteri
cell-free superants Knysh O.V., Martynov A.V., Voyda Yu.V., Babych Ye.M. Introduction.
Precursor
directed
biosynthesis is one of the promising approaches
to finding new antimicrobial agents and creating next-generation
probiotics. L. reuteri
is capable to convert triatomic polyol glycerol into reuterine, a
broad-spectrum antimicrobial substance. There
are no data on the use of other polyols as
precursors. The aim of the research
was to investigate the effect
of cell-free supernatants obtained
by culturing L. reuteri DSM 17938 in
its own disintegrate, supplemented with polyols (xylitol, sorbitol,
mannitol
and glycerol & glucose) on the daily biomass growth of opportunistic microorganisms.
Material
& methods. Reference
strains Staphylococcus aureus
ATCC
25923;
Escherichia coli AТСС
25922 and Pseudomonas
aeruginosa
clinical
isolate were used as a test
cultures. The
effect of the
lactobacillus supernatant 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). The final concentration of supernatants in the incubation medium
was 30%, and the final concentration of
bacterial cells was ~106
CFU/ml.
Inhibition (II) or
stimulation (SI) indices
of
the daily biomass growth of test cultures by formula were calculated.
Results & discussion.
Supplementation of culture
medium with
glycerol & glucose during L.
reuteri
cultivation resulted
in the S. aureus
(II
= 70.7%), Escherichia coli (II =
72.2%) and P. aeruginosa (II = 74.7%) daily biomass growth inhibition. As a result
of L. reuteri cultivation in its own
disintegrate supplemented with mannitol, the supernatant acquired growth-promoting
properties with respect to S. aureus (SI = 45.5%), E. coli (SI = 19.1%) and P. aeruginosa
(SI = 19, 9%). The supernatant obtained
after L. reuteri cultivation in
disintegrate supplemented with sorbitol had no significant effect on
the S. aureus and Escherichia coli
daily biomass growth, but significantly stimulated the growth of P.
aeruginosa (SI = 29.4%). The supernatant of L. reuteri,
cultured in disintegrate supplemented with xylitol had
no effect on staphylococcus growth, inhibited of E. coli
(II = 16.5%) growth and increased of P. aeruginosa (SI
= 19.1%) daily
biomass growth. The data obtained for glycerol, the
introduction of which into the culture medium of L.
reuteri led to the appearance of inhibitory activity of the
supernatant against all test cultures, were expected. They coincide with the results of studies by
other authors and are associated with the ability of this type of
lactobacilli
to convert glycerol into a broad-spectrum antimicrobial substance
reuterin. The results of the study
confirm that xylitol, sorbitol and mannitol do not undergo fermentation
with
the formation of acidic end products during the cultivation of L. reuteri. These polyols remain either
unchanged or undergo slight modification in the composition of the
supernatant
and have different effects on the daily
biomass growth of test cultures. Conclusion.
The
results of the study
showed that the use of xylitol, sorbitol and mannitol as precursors,
and L. reuteri
DSM 17938 as a biotransformer system in the development of new
antimicrobials
using a precursor-directed biosynthesis strategy is ineffective. They also confirmed that
the supernatant obtained after cultivation of L. reuteri DSM 17938 in its own
disintegrate supplemented with glycerol & glucose, has a pronounced
inhibitory activity against the investigated opportunistic
microorganisms. Keywords: polyols, Lactobacillus reuteri, cell-free, supernats
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27-31 |
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Semchenko K. V., Vyshnevska L. I. Introduction. Troches are very popular in children due to their high
compliance,
pleasing organoleptic characteristics and easiness of administration.
Nevertheless, the expanding range of drugs in this dosage form still
lack
anthelmintic products. In order to
introduce the new anthelminthic drug in the form of chewable troches
under the
conditional name “Albenpast”, there were conducted the studies over
selection
the optimal gel-forming agent and flavorings as correctors of taste
characteristics. The purpose of this work is to develop the technology
of
chewable troches under the conditional name “Albenpast” and to
establish the
critical points of the production process. Materials & methods. As the objects of research the pure substances
(albendazole, gelatin,
glycerol, purified water, glucose syrup, fructose, citric acid, fruit
flavoring, food coloring) and samples of troches on their basis were
used. The
research methods used are reflected in the State pharmacopoeia of
Ukraine. Results &
discussion. Research was conducted against
the chewable troches “Albenpast” to the composition of the components
of which
the patent is claimed. Compositions of troches 1 and 2 differ in the
type of
sweetness flavoring: 1 contains glucose syrup, 2 contains fructose.
Composition
2 is offered for the use in children who need to control the level of
glucose. The
production process offered includes 9 stages with critical points in 2,
3, 4, 5
and 7. Additionally, for each stage the possible types of quality
control are
described. The further quality control of the obtained by the
offered technology samples of chewable troches showed their full
correspondence
with the requirements of SPhU. Conclusion. The technological process of production of chewable
troches is offered
and the technological scheme of their production is given. Critical
parameters
of the production process and their values are identified and
described. The
results of the quality evaluation of obtained chewable troches
according to the
main quality indicators in accordance with the requirements of SPhU are
presented. Keywords: oromucosal drug, production, helminthiases, children
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32-35 |
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Molecular
dynamics study of the structural role of metal atoms in the urease active site Lisnyak Yu. V., Martynov A. V. Introduction. Urease is a representative of a small group of enzymes
that can bind
different alternative metals to execute the same catalytic function.
The
experimental X-ray studies conclude that a urease activity critically
depends
on the precise positions of amino acid ligands at a metallocenter, the
bound
solvent molecules and the type of metal, and very subtle changes of
metallocenter structure can essentially influence the urease activity.
Are
these conclusions valid in the case of the urease structures in the
solution?
By molecular dynamics simulations, we studied these aspects for urease
derivatives with alternative metals in solution under physiological pH
and
temperatures. Methods. Molecular dynamics (MD) simulations were carried out for the following systems: Ni-containing Sporosarcina
pasteurii
urease
both native
(PDB code
2ubp)
and in complex with competitive inhibitor acetohydroxamic acid,
AHA
(PDB code
4ubp);
Klebsiella aerogenes
urease both
Ni- and Mn-containing (PDB codes
1fwj and
1ef2,
respectively);
Fe-containing Helicobacter mustelae urease
(PDB
code
3qga)
(table
1). As well, there was studied an apoenzyme of Ni-containing
urease: the structure of the complex of competitive inhibitor
AHA with Sporosarcina
pasteurii urease (4ubp) from which there were removed Ni atoms. The systems studied
(α subunits or its complex with competitive inhibitor)
were placed in a
cubic periodic cell filled with TIP3P water molecules. The
simulation cell was 1
nm larger than the molecular system studied along all three axes. Na+
and Cl¯
counterions were added to neutralize the system and to reach ion mass
fraction
0.9% NaCl. Before simulations the systems were energy-minimized. After
a short
steepest descent minimization, the the procedure continued by simulated
annealing minimization. AMBER14ipq force field was used. To treat long-range
electrostatic interactions
the Particle Mesh Ewald algorithm was used. The equations of the
movement were
integrated by 2.5 fs step. To speed up the calculations the non-bounded van der Waals and electrostatic forces were evaluated only each second step and added with the scaling factor 2. The molecular dynamics
simulations
were run in NPT ensemble at pH 7.4 and two temperatures (298 K and 310
K). Trajectories were computed for 50 ns,
the
data were saved each 25 ps. Models
building, structure refinement, molecular
dynamics simulations,
and analysis as well as the result presentation by using molecular
graphics
were performed by
using the molecular
modeling program YASARA
Structure. Results and discussion. After
equilibration the RMSD values for different systems are close to each
other and
change insignificantly (except Fe-containing Helicobacter mustelae urease), evidencing that their global structure is quite stable
and that no
significant conformational
transformations occur within these systems, whereas H. mustelae urease structure revealed instability during the
simulations. The presence of the competitive inhibitor in the active
site did not change
the Ni
(1) and Ni
(2) coordination numbers. There were observed no essential
deformations of the
geometry of the ions binding with the active site ligands as well. A
similar
situation was observed in the case of Mn-containing K.
aerogenes
urease too. In the case of S. pasteurii urease
apoenzyme,
there were observed the insignificant shifts of the active site
residues. The
root mean square deviation of
the residues of the
active site of apoenzyme relative to holoenzyme was 0.65 Ǻ. In the absence of Ni ions in
apoenzyme, the position
of the inhibitor AHA within the active site is unstable and it
gradually drifts
and leaves the active site. For all ureases, the temperature increase
from 298
K to 310 K had a little effect on the average distances “metal-ligand”.
The
temperature increase from 298 K to 310 K had an insignificant effect on
the
distances “metal-water” in the active site of the Ni-containing ureases
S.
pasteurii and K. aerogenes and a little influence on these
distances
in the Mn-containing K. aerogenes urease. The metallocentre
structures
in the Mn- and Ni-containing K. aerogenes ureases are very
similar. Conclusions. There have been studied the
structural role of the nickel ions in a urease active site, the
influence of the
temperature and the ion type on the structure of the urease active
site. It
have been shown that binding of the competitive inhibitor
(acetohydroxamic
acid, AHA) did not change the Ni ions coordination in the urease active
site
and did not essentially effect the
geometry of the active site near the nickel ions. The main factor of
the
inhibitor binding are the nickel ions. It have been shown that the
active site
structures of the Ni- and Mn-containing ureases Klebsiella aerogenes and Sporosarcina pasteurii are approximately
identical. It have been shown that the metallocentre structure of these
ureases
are in general stable regardless of the urease source, the ion type and
the
temperature. Keywords: urease metallocentre, Ni-, Mn- and Fe-containing urease, molecular dynamics.
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6-48 |
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Standardization of a method for identification of
elecampane
sesquiterpene lactones |
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49-53 |
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Research of antimicrobic activity of foaming products
samples with octopirox Zaika S.V., Strilets O. P., Baranova I.I., Bezpala Yu. O.,
Martyniuk Т. V. Introduction. Seborrheic dermatitis
(SD) - chronic recurrent skin disease, which is associated with
increased
cutaneous fatty secretion, change in its qualitative composition and is
characterized by localization in the areas of accumulation of sebaceous
glands
on the scalp, face, upper torso. Therefore, the choice of tactics for
the
treatment of patients with SD depends on the degree of clinical
manifestations,
duration of the disease, information about the effectiveness of
previously
conducted therapy. Particular attention is paid to the choice
of
antifungal medicines in the treatment of SD. Traditionally,
local remedies in the form of ointments or creams are used to treat SD
of the
scalp, they cause many inconveniences when used. In this respect,
foaming
products, in particular shampoos, which include active substances that
have
certain therapeutic potential regarding the main mechanisms of SD
treatment,
having self-regulating, anti-inflammatory, antifungal, antibacterial,
reparative, moisturizing properties, etc. are very appropriate. One
of
the main factors in the development of this disease is the yeastlike
fungus of
Malassézia fúrfur genus (or Pityrósporum ovále), which is present in
the skin
of each person. Octopirox®, which has strong fungicidal (antifungal)
and
antibacterial actions, was chosen among a number of modern products in
this
field. Therefore, the main purpose of our experiment was to study the
antimicrobial activity of developed samples of foaming agents with
octopirox to
justify the rational concentration. Materials and methods.
For this study, we produced a number of experimental samples
of foaming
bases with octopirox and α-lipoic acid at different concentrations: sample № 1
- foaming base; sample № 2 – foaming
base + α-lipoic acid; sample № 3 – foaming base + octopirox (0,25
%); sample № 4
– foaming base + octopirox (0,5 %); sample № 5 – foaming base +
octopirox (0,75
%); sample № 6 – foaming base + octopirox (1,0 %). The antimicrobial activity of prototype gels was studied in vitro by
diffusion in agar (“wells” method). This method is based on the ability of active substances to diffuse in the agar medium, which was previously inoculated by bacterial crops.
The
results of the studies make it possible to characterize both the antimicrobial
activity of the samples and the release of antimicrobial substances from the base,
because the growth inhibition zones of microorganisms are formed as a result of the
diffusion of
these substances into a dense
nutrient medium.
Results. Based on the experimental
data, in the development of the foaming product composition with
octopirox AFI
concentration of 0.5% is optimal and further increase it to 0.75%
(sample No.
5) and 1.0% (sample No. 6) is impractical. It should be noted that the
studies
showed that foaming samples had high antifungal activity and moderate
effect on
gram-negative bacteria, so in the development of the optimal
composition of the
tool and its long-term storage and use, it is necessary to consider the
introduction of preservatives and conduct appropriate research. Conclusion.
Thus, the results of the experiments showed that the test samples No. 1
and No.
2 did not have any antimicrobial action pertaining to gram-positive,
gram-negative bacterial cultures and antifungal activity. Samples No.
3, No. 4,
No. 5 and No. 6 of the foaming product with octopirox at concentrations
of
0.25%, 0.5%, 0.75% and 1.0% have a broad spectrum of antimicrobial
action
pertaining to gram-positive (Staphylococcus
aureus ATSC 25293 and spore cultures of Bacillus
subtilis ATSS 6633), gram-negative (Escherichia
coli ATSS 25922 and Pseudomonas
aeruginosa ATCC 27853) bacterial cultures, as well as in relation
to the
cultures of Candida albicans fungi
ATSS 885-653 and Aspergillus niger fungus
ATCC 16404. However, the experimental results showed that sample No 4
(octopirox concentration of 0.5%) was the most promising for further
work on
the development of antimicrobial activity of the foaming product. Key words: seborrheic dermatitis, antimicrobial activity,
octopirox, foaming product, product for men. |
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