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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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2-7
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ÎÃËßÄÈ
(REVIEWS)
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TUBERCULOSIS AS
AN INFECTIOUS PATHOLOGY OF IMMUNE SYSTEM
Martynov A.V.,
Bomko T.V., Nosalskaya T.N., Lisnyak
Yu.V., Romanova E.A., Kabluchko T.V., Sidorenko T.A., Igumnova N.I.,
Pogorelaya
M.S., Shcherbak E.M., Yukhimenko V.I.,
Farber B.S., Farber S.B.
http://doi.org/10.5281/zenodo.167423
As a result of
years’ research of the many research groups around the world able to
understand
the reason why it will be impossible to create really effective vaccine
for the
prevention of tuberculosis infection in the near future. The main
reason for
the impossibility creating such vaccine is an intracellular nature of
tuberculosis. In fact, TB is a pathology of the immune system.
Mycobacterium tuberculosis
persist within macrophages and thereby inhibit the process of
phagocytosis
completion and digesting the contents of phagosome. The destruction of
the
lysosomal membrane inside macrophages is blocked by changing the pH in
lysosomes. For the presence of lytic activity for most lysosomal
enzymes
require need acidic environment. Mycobacteria are also getting into the
lysosomes of macrophages start to rapidly hydrolysis for urea by urease
to form
ammonia. Wherein pH in the medium changes to alkaline, this inactivates
enzymes
and stabilizes lysosomal membrane. Thus mycobacterium prevent lysosome
collapse
at inactivated lysosomal enzymes and do not allow them to complete
macrophage
digestion phase by transition lysosomal to phagosomal stage. Stop
phagocytolysis
process leads to imbalance of the host immune system. Increasing the
number of
infected macrophages sensitized to Mycobacterium tuberculosis antigens,
leading
to constant hyperfunction of cellular immunity, particularly enhanced
immune
response to cell wall components of mycobacteria, induction high titers
of
interferon-gamma in response to a stimulus, a sharp jump IL-2 titers
and TNF-α
, IFN-γ specific activation CD8 + CTL. Need also focus
attention on the main differences from the MBT and human BCG, that is
well
growth in the human body, persists along host life, but does not cause
active
TB (except in patients with HIV/AIDS). After MBT cell destruction in the environment gets some additional
high
allergenic antigens, such as 85B, ESAT6, Rv2660c, HyVaC 4 (Ag85B and
TB10.4.).
These antigens to provide high adhesion and allergenicity of human
strains M. tuberculosis. Most allergens
that cause
obvious signs of active tuberculosis are the antigens ESAT6 and CFP10.
Such
protein antigens can be called endotoxins. Also to pathogenicity
factors
include cord-factor, it main component is a polysaccharide-mycolic
complex from
cell wall (Figure 2) containing ftiolic and mycolic acid - to ensure
the
stability of mycobacteria to lysosomal enzymes. Currently
available
diagnostic tools tuberculin preferably contain the above components of
the cell
wall and differences (from BCG) allergens ESAT6 and CFP10. Currently
well
established that the virulence of M. tuberculosis, mainly responsible
genes
encoding antigens ESAT-6 and CFP10. When comparing the genomic sequence
of M.
tuberculosis with attenuated M. bovis
BCG was detected genomic deletion of the three sites in the vaccine
strain
(RD1, RD2, RD3). BCG vaccine strain genome stripped areas in the RD1,
encoding
mycobacterial antigens ESAT-6 and CFP-10 present in virulent strains of
M.
tuberculosis. Many researchers believe that mutations in genomic
regions RD1,
encoding mycobacterial antigens ESAT-6 and CFP-10, occurred in the
process of
creating a BCG strain. It remains not examine the question of whether
all
strains of M. bovis other than BCG have antigens ESAT-6 and CFP-10, and
whether
they depend on the degree of virulence of the mycobacteria strains.
About a
third of the population is infected with the MBT. Tuberculosis
statistics show
that out of every 100 man infected MTB, only 10 appear open clinical
forms. In
the remaining patients, positive skin test and/or gamma-interferon
test,
clinical symptoms of tuberculosis never does
occur, and no signs of sensitization other than to MBT
antigens and
presence ESAT6 - antibodies in the blood. Thus, if the focus is not on
the
infection, but on the prevention of tuberculosis reactivation, can
significantly reduce the number of cases with clinical manifestations.
There
have been recent publication comparing the immunity of patients with
open
clinical forms tuberculosis and without clinical symptoms, but ESAT6 -
test-positive. One of the rational ways for helps to MTB - infected
macrophages
is the simultaneous use of urease inhibitors and simultaneously use
selective
activators of antibacterial complete phagocytosis. For the latter
group, some
authors include also histone deacetylase inhibitors (HDAi). The use of
such
inhibitors in the latter case will mass increase number reading frames
in the
macrophages genome and leads to stormy expression phagocytosis
activators, that blocked by MBT.
These inhibitors include valproic acid and trichostatin. Research in
this area
only started, and the expectations are very high. Another activator
phagocytosis with very similar action mechanisms is the vitamin D3 -
ergocalciferol. In a variety experiments shows that the soluble
derivatives of
vitamin D3 inoculation to the culture of MBT - infected macrophages
leads to
the completion phagocytosis and complete digestion of the MBT. The
disadvantage
of this method is the need to maintain a concentration of vitamin D3,
which is
quite toxic to the human body as a whole. Accordingly, a new
form
vitamin D3 is to be administered directly to the places where many
infected
macrophages, i.e. as an aerosol through the lungs. Also pay attention
to the
fact that, earlier for purpose combating tuberculosis the urease
inhibitors
have not been used, although quite a lot of well-known non-toxic
compounds anti
-urease activity. Thus, the most promising way to prevent tuberculosis
reactivation in humans with positive test specimens and humans in
remission
following chemotherapy is to provide an aerosol preparation containing
both
urease inhibitor, activator phagocytosis vitamin D3 and histone
deacetylase
inhibitor. The use of such aerosol once a week will greatly reduce the
number
of macrophages with incomplete phagocytosis and prevent the background
to
tuberculosis with clinical open forms. This disease, like tuberculosis,
prevention is better than cure, especially with the emergence of M.
tuberculosis multiresistant strains.
Keywords: tuberculosis,
immunity, phagocytosis, vaccines
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8-14
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ÐÎËÜ ËÅÉÊÎÇÍί ÑÒÎÂÁÓÐÎÂί Ê˲ÒÈÍÈ Ó ÏÀÒÎÃÅÍÅDz ÕÐÎͲ×Íί ̲ªËίÄÍί ËÅÉÊÅ̲¯
Ñâºæåíöåâà ².Î.
ROLE OF LEUKEMIC STEM CELLS IN THE
CHRONIC MYELOID LEUKEMIA PATHOGENESIS
Sviezhentseva I.O.
http://doi.org/10.5281/zenodo.167425
The presence of leukemic stem cells
(LSC) in the bone marrow of patients with chronic myeloid leukemia
(CML) is the
cause of relapses as a result of the treatment with chemotherapeutic
agents and
target therapy drugs. This is due to the ability of LSC to attach
itself to the
microenvironment cells and to remain at rest for a long time. Vascular
and
osteoblasts niche play
a very important role in this process. However, for
being in G0 phase LSC have direct contact with the cellular elements of
bone
marrow microenvironment. So LSK contact with mesenchymal cells of bone
marrow
using the appendixes, connecting components invaginations and lint. The
cadherins and integrins are important in the interaction of osteoblasts
niche.
They are able to activate intracellular signaling cascades that provide
resting
state of LSK. In addition, a bone marrow niche provides changes of LSC
oxidative metabolism, which also plays an important role for cell entry
into
the G0 phase. Further, LSC also have certain physiological properties,
which
play an important role in the drug resistance formation, particularly
drugs
with targeted actions - tyrosine kinase inhibitors. LSK characterized
by a high
level of BCR-ABL expression and their population can have a lot of
point
mutations in the bcr-abl gene in the
same patient. This leads to the fact that the taken medicines dose does
not act
against LSK, reducing the number of a whole leukemic cells clone.
However,
complete LSC elimination from the the patient’s bone marrow need search
the
main differences between the LSC and normal HSC. After the literature
analysis
it was found that LSC have several significant differences such as the
ability
to cause leukemia during the transplantation to immunodeficient
animals, this
leukemia is morphologically and phenotypically similar to the original
tumor,
in addition the LSC can be transmitted from animal to animal. In
addition, the
LSC is also characterized by the mutations presence in the genes of
kinase
domains, transcription factors and tumor suppressors (genes of growth
factors
FLT3, C-KIT, genes K-RAS and N-RAS, mutations in genes STAT5A, TP53,
AML1, RB1,
MYC, p16 / NK4a, ENV1).
Now the most important role in LSK
biology research takes studying of signaling cascades involved in the
processes
of cell activity. This key molecule of cell signaling pathways can
become
targeted agents that may be used for the elimination of LSC from the
patient
bone marrow. However, it is necessary to distinguish the molecular
cascades
that are inherent to all bone marrow stem cells from LSC specific
intracellular
signal transmittors. Common to the LSC and HSC are the following
signaling
pathways: Wnt/β-catenin, Sonic Hedgehog and Noch signaling pathways.
Moreover,
there are signaling cascades that are specific only for LSC. They are
charecterized by the exclusive expression of Alox5, AHI-1 and NFκB
genes. In
addition, the LSC are also characterized by the increased expression of
AVSV-1
and ABCG-2 transporters, providing the evacuation of the cell
chemotherapeutic
drugs. LSC are characterized by the decreased expression of Oct-4,
which ensures
the supply of drugs to cells. The article also highlights the key therapeutic
tactics that can be used to eliminate the recurrence of CML associated
with the
presence of LSC, which remain at rest for a long time, in the bone
marrow of
patients. The first tactic is elimination of LSC using the targeted
drugs that
operate solely on the target molecule in leukemic cells. The second
approach is
a direct administration of drugs to a patient that could promote a
permanent
state of rest for LSC in order to prevent relapses.
Keywords: leukemic stem
cells, chronic myeloid leukemia, hematopoietic stem cell
microenvironment.
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15-21
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ÑÒÐÓÊÒÓÐÍÀß
ÎÐÃÀÍÈÇÀÖÈß ÁÀÊÒÅÐÈÀËÜÍÛÕ ÓÐÅÀÇ
Ëèñíÿê
Þ. Â.,
Ìàðòûíîâ À. Â.
STRUCTURAL ORGANIZATION OF
BACTERIAL UREASES
Lisnyak Yu. V., Martynov A. V.
http://doi.org/10.5281/zenodo.167426
This brief review concerns the basic
principles of structural organization of multi-subunit bacterial
ureases and
formation of their quaternary structure.
Urease is a nickel-containing enzyme (urea
amidohydrolase, ÅÑ 3.5.1.5) that catalyses the hydrolysis of urea to get
ammonia and carbamate which then decomposes with water to get ammonia
and
carbon dioxide. Urease is produced by bacteria, fungi, yeast and
plants. On the
basis of similarities in amino acid sequences, ureases assumed to have
a
similar structure and conservative catalytic mechanism. Within past two
decades
bacterial ureases have gained much attention in research field as a
virulence
factor in human and animal infections. The first crystal structure of
urease
has been determined for that from Klebsiella
aerogenes. The native enzyme consists of three subunits, UreA
(α-chain),
UreB (β-chain) and UreC (γ-chain), and contains four structural
domains: two in
α-chain (α-domain 1 and α-domain-2), one in β- and one in γ-chain.
These three
chains form a T-shaped heterotrimer αβγ. Three αβγ heterotrimers form
quaternary complex (αβγ)3. In case of Helicobacter
pilori, the analogous trimers of corresponding dimeric
subunits (αβ)3 form tetrameric structure ((αβ)3)4
in which four trimers are situated at the vertexes of the regular
triangle
pyramid. Active center is located in α-domain 1 and contains two atoms
of
nickel coordinated by residues His134, His136, carboxylated Lys217, His
246,
His272 and Asp360, as well as residues involved in binding (His219) and
catalysis (His320). Active site is capped by a flap that controls
substrate
ingress to and product egress from the dinickel center. Urease requires
accessory proteins (UreD, UreF, UreG and UreE) for the correct assembly
of
their Ni-containing metallocenters. The accessory proteins UreD, UreF,
and UreG
sequentially bind to the apoprotein (UreABC)3 to finally
form
(UreABC-UreDFG)3 activation complex. UreE metallochaperone
delivers
nickel ions to this complex, UreE and (UreDFG)3 are then
released
from the activated enzyme. An
understanding of
structural organization of bacterial ureases is the necessary factor in
the
studies of structure-function relationships of these enzymes,
mechanisms of
their enzyme and non-enzyme activity, in design of new safe and
efficient
enzyme inhibitors aimed to struggle with infectious diseases promoted
by urease
activity.
Key
words: urease, sub-unit organization, quaternary structure,
accessory proteins
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22-30
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ÂËÈßÍÈÅ
ÃÅÐÏÅÑÂÈÐÓÑÍÎÉ ÈÍÔÅÊÖÈÈ ÍÀ
ÈÌÌÓÍÍÓÞ ÑÈÑÒÅÌÓ È ÈÌÌÓÍÍÛÉ ÎÒÂÅÒ ÎÐÃÀÍÈÇÌÀ ÍÀ ÂÀÊÖÈÍÀÖÈÞ
Âîëÿíñêèé
À. Þ., Êó÷ìà Ì. Â.,
Êîëîòîâà Ò. Þ., Êëûñà Ò. Ë., Êó÷ìà È. Þ.,
Êîíîðåâà
Å. Ñ., Ñìåëÿíñêàÿ Ì. Â.,
Ïåðåìîò Ñ. Ä., Êàøïóð Í. Â., Äàâèäåíêî Ì. Á.
THE IMPACT
OF PERSISTENT HERPESVIRUS INFECTION ON IMMUNITY AND VACCINATION RESPONSE
Volyanskiy A. Yu., Êuchma M. V., Kolotova T. Yu., Êlyisa
T. L., Êuchma I.
Yu., Konoreva Å. S., Smelyanskaya Ì. V., Peremot S. D., Êàshpur N. V.,
Davydenko Ì. B.
http://doi.org/10.5281/zenodo.167428
In this review we summarize current knowledge on
the ability of latent herpesviruses to modulate the
immunity and response
to vaccination. Nearly all humans are latently infected with multiple
herpesviruses but little is known about virus-host interactions.
Meanwhile, the
study of the immune response to Epshtein-Barr virus (EBV) and ñytomegalovirus (CMV) has revealed significant regulatory effects on the
immune system. During the primary infection a human cytomegalovirus is
predominately found in peripheral blood monocytes and polymorphonuclear
leukocytes. However, the virus can
not be
replicated in these cells. CMV induces the
survival and differentiation of infected monocytes into long-lived
macrophages
capable of supporting viral replication and the release of virions,
which infect CD34+ myeloid progenitor cells. CMV latently
persists in myeloid progenitor cells and monocytes and reactivates
during their
differentiation into macrophages. CMV-infected
monocytes exhibit a unique reprogramming of their differentiation and secret both pro-inflammatory M1- and
anti-inflammatory M2-associated cytokines. But
cytomegalovirus induced macrophage
phenotype skewed towards pro-inflammatory M1 type. MV has profound effects on the composition and function of
both T cells
and NK cells. CMV constantly reactivates during
differentiation of monocytes into macrophages.
Consequently, persons with latent CMV infection have
substantially increased numbers and proportions
of CD8+ T cells that lead to exhaustion and an
early onset of immunosenescence. Also, it has been shown that the latent CMV virus infection markedly
increases the proportion of NK cells expressing
the activating NKG2C receptor. So, it has been proposed that CMV alters the composition of T cell and
NK cell
subsets and accelerates immune aging. Given the capacity of CMV to
alter a macrophage, as well as NK
and T cell
responses it is reasonable to hypothesize that latent infection
would alter the
outcome of vaccination. The EBV virus remains in
memory B cells throughout life. In
healthy subjects
the EBV remains latent in the latency phase 0
and
EBV replication proceeds without
production of
infectious virions. But the virus
can be
reactivated in latency phases 1, 2 and 3. The
virus reactivation can affect immunity and results in diseases. Chronic
fatigue syndrome (CFS) is characterized by fatigue, exhaustion and
flu-like
symptoms. EBV latent reactivation in CFS patients is
supported by certain data. In a subset of patients, CFS begins with infectious mononucleosis
and enhanced EBV-specific
antibody titers have been reported. Also, a
profound deficiency in EBV-specific B and T cell memory response was
observed
in a majority of CFS patients. These
data
confirmed the EBV virus involvement in the CFS development. Cytokine
dysregulation, decreased natural killer cell functioning, the presence
of
autoantibodies, and a reduced response of T cells to mitogens have been
reported in CFS. But if immunity is
disturbed
in CFS patients, they might have an altered response to vaccination. Herpes
viral reactivation has been documented in sepsis. Demonstration
of the widespread reactivation of latent herpesviruses in sepsis
provides
strong evidence that sepsis results
in
functional immunosuppression. Reactivation
of
latent viruses may be associated not only with sepsis but with aging as
well. Moreover, according to our
data herpesvirus
reactivation is common in recurrently infected children. These
observations highlight the ability of herpesviruses to profoundly
impact the
host immune function. But the recent
publications have shown that persistent herpesvirus infection can be
beneficial
for the host. The data obtained from
the
multiple mouse models demonstrate the potential for herpesvirus
infection to
enhance resistance against a
secondary
infection. It has been documented
that during latent murine
cytomegalovirus or murine
gammaherpesvirus infection 68 mice
are
protected against lethal bacterial infections by prolonged macrophage
activation and IFNγ secretion. Little is known about the potential benefits of
herpesvirus latency in
humans. The effect of CMV on vaccine responses is controversial. It is well known that vaccine
responses is reduced in aging populations. CMV
accelerates immune aging and may
further reduce the response to
vaccination.
In fact, some studies show a negative effect of CMV while
others showed
no difference in CMV+ vs CMV− in older individuals. Conversely, in
young
individuals, a negative association between the CMV antibody titer and
the
response to the influenza vaccine has been found. Also,
a negative association between the CMV antibody titer and the response to
the
influenza vaccine was found in young individuals. But, according to
another
results CMV enhances the immune responses of younger individuals to
influenza
vaccination. In summary, the
hypothesis that
CMV virus accelerates immunosenescence and decreases vaccination
response is
controversial. Moreover, CMV
infection may
enhance the immune responses in children and young individuals. Vaccinations
can
induce the aberrant immune response of CFS. But
the available data do not support the CFS impact on the vaccination
response. In conclusion, the host-persistent
herpesviruses infection history is likely to play a significant role in
the
immune system response to vaccination.
Key
words: latent herpesvirus infection, cytomegalovirus, virus Epshtein-Barr, chronic fatigue syndrome, T cells, NK cells, viremia, vaccination.
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31-44
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ÅÊÑÏÅÐÈÌÅÍÒÀËÜͲ
ÐÎÁÎÒÈ
Experimental papers
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ETIOLOGIC
FACTOR IN THE DEVELOPMENT OF MYOCARDITIS IN THE KHARKIV REGION
Smelyanskaya M. V., Peremot S. D., Kashpur N. V.
http://doi.org/10.5281/zenodo.167429
Introduction.
According to the
research conducted in the last decade, there has been growth in the
non-coronary disease infarction among all cardiovascular diseases. The
prominent place among all non-coronary heart diseases is taken by
myocarditis,
which predominantly affects young people of working age (30-40 years
old).
According to the bibliography, the prevalence of myocarditis is 20% of
the
non-coronary heart lesions and by different authors 5 - 11% of the
total amount
of diseases of the cardiovascular system. To date, there are no clear
criteria
of infectious myocarditis. It is widely accepted that myocarditis is
natural
complication of infectious diseases in which any infectious agent may
be the
etiological factor. Until recently, Coxsackie virus of group A, B was
considered
the most cardiotropic. However, the role of enteroviruses has been
recently
reviewed in favor of persisting viruses and especially family
Herpesviridae. Optimization
of
myocarditis diagnosis using noninvasive tests, will not
only reveal the true extent of the disease but may
also enable to examine
viral
myocarditis as a much
more common
pathology than it seems at present and will increase understanding of
the significance
of this pathology
in the
cardiovascular continuum.
Material
& methods.
87 people diagnosed with infectious myocarditis were
examined. Blood and other biological fluids were subject to
examination.
Patients of the main group have also had their biopsy material,
obtained in a
result of endomyocardial biopsy, and pericardial fluid, derived as a
result of
diagnostic and therapeutic puncture under hydro pericarditis, examined.
PCR was
performed to determine the genomic sequence of enterovirus (HEV),
adenovirus
(HAdV), human cytomegalovirus (CMV), herpes simplex virus (HSV),
Epstein-Barr
virus (EBV), human herpes virus 6 (HHV6) and influenza A viruses and B.
Results &
discussion. Our research confirms the global trend of reducing the
role of
enteroviruses in infectious myocarditis. In
the course of this investigation, herpesvirus markers in
biological material from patients in the form of monoinfection or mixed
infections were found in 67 patients with myocarditis, which amounted
to 87%.
While in the control group, similar markers have been found only in 9
patients
or in 22.5% of cases. Herpesvirus detection rate is almost 7 times the
allocation of other viruses in infectious myocarditis. Different types
of
herpesvirus with varying frequency were found in the main (SM and CM)
and
control groups. According to our data antigens of CMV, HHV6 were found
in
60-70% of patients suffering from subacute myocarditis, and almost 40%
of
patients in this group had markers HSV1,2 and VZV. In patients with
chronic
myocarditis the percentage of CMV and HHV 6 markers identification
reached
70-75%, HSV1,2 - exceeded 50%, EBV - 31,4%. While VZV markers did not
much
exceed the performance of the control group (16.2% and 12.5%
respectively).
Determining markers of enteroviruses in the study groups and the
control group
were not significantly different. Our
data detection of mixed infections indicates a very significant (75%)
share of
the combination of five different herpesvirus in patients with
myocarditis. It
is predominantly a combination of HHV6 with CMV, HSV1,2, EBV and VZV.
The data
can be used not only to choose the etiotropic
treatment of myocarditis, but also as a diagnostic criteria when
combined with
a history of clinical indicators. In comparison of herpesvirus DNA
detection in
patients with SM in blood and EMBS, it was revealed that markers of CMV
and
HHV6 were almost equally met both in blood and in heart biopsies.
Viruses
HSV1,2 and VZV were more often detected in the blood than in the EMBS
(3 and
1.5 times respectively). This gives reason to believe that the
identification
of herpesvirus markers in the blood of patients with AM can be used for
non-invasive etiological diagnosis of subacute viral myocarditis.
Development
and introduction into medical practice of new non-invasive methods of
myocarditis diagnosis and their etiology specification will enable to
examine
viral myocarditis as a much more common pathology than it would seem. Conclusion.
During virological examination of biological material of
myocarditis
patients, a high proportion (87%) of antigens of Herpesviridae viruses
were
revealed. In patients with myocarditis, association of herpesvirus
antigens of
various types (in 75% of detection) dominate, while in healthy people
of the
control group - mono infection can be detected more often. Chronic
myocarditis
patients in 61% of cases are detected with three or four antigens of
viruses of
the family Herpesviridae, basically HHV6 with CMV, HSV1,2, EBV; with
subacute
myocarditis – HHV6 with CMV, HSV1,2 and VZV. The findings point to the
need for
mandatory examination for Herpesviridae virus of patients diagnosed
with
myocarditis.
Keywords:
myocarditis, Herpesviridae virus, endomyocardial
biopsy.
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45-48
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NEW FORMYL-PEPTIDES, AS
STIMULATOR OF NON-SPECIFIC ORGANISM RESISTANCE AGAINST MYCOBACTERIA
Pohorila M.S., Martynov A.V.,
Romanova O.A., Sidorenko T.A., Igumnova N.I.,
Yukhimenko V.I., Shcherbak O.M.
http://doi.org/10.5281/zenodo.167430
Introduction. The
key element in the formation of tuberculosis infection (TI) is the
inability of
alveolar macrophage to complete phagocytosis of mycobacteria absorbed
by them,
that caused by both features of the pathogens biology and the tissue
macrophages. It is known, that M. tuberculosis is capable long-term
persistence
and proliferation in alveolar macrophage cytoplasm because of high
stability of
them cell walls to the lysosomal enzymes action. Mainly, the phenomenon
of
granulomatous reaction, inherent of tuberculosis (TB), reflects the
inadequacy
in elimination of tuberculosis pathogen of alveolar macrophages. Thus,
the
inclusion of agents that can activate completed phagocytosis of
mycobacteria by
alveolar macrophages, in the base of anti-TB therapy is a promising
direction
in the prevention of latent tuberculosis reactivation. Materials
and Method.
The ability of formyl-peptides activate the completeness of
mycobacteria
phagocytosis by alveolar macrophages absorbed by them were evaluated in
vitro.
For reaching the aim of the study we had used a broncho-alveolar lavage
obtained by white laboratory male mice 2 months of age. As a comparison
drug we
used the officinal preparation "Liasten". To determine the lysosomal
activity by the presence of peroxidase was treated with acridine orange
causing
selective staining red lysosomes. Acid phosphatase activity was studied
using
azocoupling reaction for staining granules in the cytoplasm blue or
purple. Results
are expressed as mean cytochemical coefficient (LZC). Results and
discussion. Incubation of alveolar macrophages and formyl-peptides
leads to
a significant increase the index of mycobacteria phagocytosis
completeness for
vaccine strains - (1,70 ± 0,31) and (1,20 ± 0,22), respectively, (p
<0.05).
The standard medication - Liasten - also increased the "killing"
ability of tissue macrophages compared to control: (1,8 ± 0,25) and
(1,20 ±
0,22), respectively, (p <0.05). Lysosomal activity of alveolar
macrophages
exposed formyl-peptides significantly increased in comparison with the
control
- (97,80 ± 5,1) and (80,9 ± 4,3) acridine orange-positive cells,
respectively,
(p <0.05). However, the effect of formyl-peptides on lysosomal
activity of
macrophages did not exceed the reference drug action - (97,80 ± 5,1)
and (95,72
± 5,3) acridine orange-positive cells, respectively, without
significant
differences. Incubation with formyl-peptides resulted in a significant
increase
of LZC of acid phosphatase in macrophages - (2,08 ± 0,20) and (1,59 ±
0,14),
respectively, (p <0.05). Significant differences between the content
of this
enzyme in macrophages when exposed formyl-peptides and the reference
drug were
not detected. Conclusion. As a result the co-incubation of
alveolar
macrophages and new formil-peptides activates BCG phagocytosis
completeness.
Also, there is the influence of the studied substances under
significant
lysosomal activity and increase the content of acid phosphatase in
macrophages
isolated from broncho-alveolar lavage in comparison with the control.
The level
of functional activity stimulation of phagocytes under the influence of
formyl-peptide is the same, that we registered after the Liasten
administration. It has indicating prospects of the medicinal
preparation
creation on the formyl-peptides basis, which stimulates the organism
non-specific resistance.
Key words: Formil-peptides,
non-specific resistance, lysosomal activity, in vitro.
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49-52
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²ÍÔ²ÊÎÂÀͲÑÒÜ HHV-6 ²ÉÑÜÊÎÂÎÑËÓÆÁÎÂֲ ÕÂÎÐÈÕ ÍÀ
ÍÅÃÎÑϲÒÀËÜÍÓ ÏÍÅÂÌÎͲÞ
Áðóñí³ê Ñ.Â., Ïîïîâà Í.Ã.,
Ïîïîâà Ë.Î.
INFECTION WITH HHV-6 OF MILITARY MEN AFFECTED BY
COMMUNITY-ACQUIRED
PNEUMONIA
Brusnik S. V., Popova N. G., Popova L. O.
http://doi.org/10.5281/zenodo.167431
Human herpesvirus, 6 type (HHV-6) was isolated at the end
of the 20th
century from the blood leukocytes of patients with lymphoproliferative
diseases. Serological
studies conducted in different countries, indicate ubiquitylation of
the HHV-6
and the existence of two antigenic variants - HHV-6A and HHV-6B. Their
high
tropism is determined in vitro to lymphocytic, nervous and dendritic
cells of
the CNS. Virus replicates in many cell, primary and passaged cultures
of
different origins. The reproduction cycle of HHV-6 continues on average
4-5
days forming syncytiums and intracytoplasmic and intranuclear
inclusions.
Significant destruction and lysis almost 90 % of infected cells is
reported
after 5-10 day of monitoring. The utility of experimentation investigating the role
of HHV-6 in the development of acute and chronic diseases in
respiratory tract
is caused by the fact that many patients, particularly those with
chronic
diseases, have complaints to chronic fatigue, decreased performance and
low-grade temperature more than 3-6 months. Several studies demonstrate
the
presence of HHV-6 in saliva, salivary and bronchiolar glands, in swabs
from
pharyngonasal cavity and gorge. Tropism of HHV-6 to oropharyngeal
epithelium
with the possibility of finding the virus in the saliva and swabs from
pharyngonasal cavity and gorge was found at the end of 20th century.
This fact
gave the basis for work determining the level of infection by this
pathogen in
patients with infectious and inflammatory pathology of the respiratory
tract. Materials and methods. Serological studies were conducted with 38 soldiers
affected by
community-acquired pneumonia. Most of the surveyed patients were ranged
in age
from 20 to 45 years old, middle age (32,5±1,5) years. Patients were in stationary treatment in the
Kharkov military hospital. The criteria for inclusion in the study on
the
infection of HHV-6 were soldiers affected by community-acquired
pneumonia with
atypical course of disease. Some patients have short-term (1-2 days),
macular
papulose eruption on hands and legs, prolonged low-grade fever, chronic
fatigue. The control group included 18 apparently healthy persons.
Enzyme
immune analysis (EIA) using commercial diagnostic system
"VektoHHV-6-IgG»
was applied to determine the immunoglobulin class G (IgG) for HHV-6 in
serum
and saliva of the examined. Registration of the EIA results was
performed using
spectrophotometer StatFax 303 by determining the optical density (OD)
in
optical experimental values and control samples of blood serum and
saliva.
Result evaluation is carried out in accordance with requirements stated
in the
instructions to the test system with obligatory consideration of
anamnestic and
clinical data from the examined patients. Results and discussion. The above
data show that among the examined patients’ infection with HHV-6 was
26,3 % of
apparently healthy persons in the control group, this figure was 2
times lower
than 10,5%. In saliva of the CAP patients as well as in blood and serum
was
found IgG to HHV-6. One patient from IgG to HHV-6 control group have
been
identified in saliva and serum. The results of the research allowed
establishing the level of seropositive individuals to HHV-6 and
demonstrate the
ability to diagnosis HHV-6 infection in infectious and inflammatory
processes
of the respiratory tract. Conclusion. The studies have shown the use of saliva as an object
of study for the establishment markers of HHV-6 in patients affected by
CAP
with important advantage - non-invasive receiving of material from
patients for
laboratory diagnosis. It can be assumed that high level of infection
for
patients affected by CAP is associated with ease droplet spread of
herpesvirus
through saliva.
Key words: human herpesvirus
6 type, community-acquired pneumonia, military man.
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53-55
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ÔÀÐÌÀÖ²ß
Pharmacy
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ÊÎÍÒÐÎËÜ
ÊÀ×ÅÑÒÂÀ ÍÀÑÒÎÉÊÈ ÁÎßÐÛØÍÈÊÀ ÌÅÒÎÄÎÌ ÂÝÒÑÕ
Õîõëîâà
Å.À.
QUALITY CONTROL OF HAWTHORN TINCTURE BY HPTLC METHOD
Khokhlova K.O.
http://doi.org/10.5281/zenodo.167436
Introduction. Hawthorn tincture is one of the most used herbal drugs
at the domestic pharmaceutical market. According to the State register
of drugs
at the pharmaceutical market of Ukraine, there are 13 commercial offers
of
Hawthorn tincture from home-produced manufactures. The initial
herbal
raw materials for Hawthorn tincture are Hawthorn fruits, which are
widespread
at the territory of Ukraine. These are pharmacopoeial herbal raw
material.
Thus, 12 different species of Hawthorn fruits are included into
monograph
<Hawthorn fruits> of Ukrainian State Pharmacopoeia (SPhU) and
State Pharmacopoeia
of USSR XI ed. On the territory of Ukraine there are near 30
different
species of Hawthorn, and the quantity of species is much arises due to
its
forms and hybrids. The ‘natural variability’ of bioactive substances of
Hawthorn fruits of
the same species and possibility of usage of many different species
during
manufacturing process of herbal drugs lead to the pitfalls in
standardization
of herbal drugs in general, and Hawthorn tincture particularly, and
should be
taken in mind while development of its quality control methods. For
development
of specific and reproducible identification method, it is necessary to
ensure
the number of parameters: usage of method and equipment that give
reproducible
results; big selections of different samples; rigorously observation of
method’s procedure of implementation. The modern, automated HPTLC
method of
analysis was chosen for identification purpose. If standardize
procedure and
suitable equipment are used, the reproducible results of the method
have to be
obtained. The aim of
this paper was development of HPTLC method for identification of
Hawthorn
tincture, which could be appropriated for stability study and
establishment of
its expire date. Materials and Methods. In research 13 samples
of
Hawthorn tinctures from 8 manufactures from Ukraine and Russia were
analyzed.
These samples were manufactured in 2010, 2014, 2015 years. The research was conducted on the base of
CAMAG laboratory, Muttenz, Switzerland. Plates used:
HPTLC glass 20x10 cm, Si 60 F254, Merck, Lot: 1.05642.0001. Material used: Automatic TLC Sampler 4,
CAMAG; Twin Trough Chamber 20x10 cm, CAMAG; Chromatogram Immersion
Device III,
CAMAG; TLC Plate Heater III, CAMAG; Automatic Development Chamber ADC
2, CAMAG;
Visualizer, CAMAG; TLC Scanner, CAMAG; Filter paper for chamber
saturation,
CAMAG; Centrifuge EBA21, Hettich; Ultrasonic Bath SW 3H, Sono Swiss;
Analytical
Balance MS 205 DU, Mettler-Toledo. Chemicals
used were
pharmacopoeial quality. Reference substances used: hyperoside, USP, batch: 33520F; rutin, EDQM,
batch: A0299493; chlorogenic acid, EDQM, batch: A0290470.
For identification of Hawthorn tincture by HPTLC the
flavonoids were chosen as a group of bioactive substances. The method
was
developed using format and style of description, which are used for TLC
Identification method for Crataegi fructus in European Pharmacopoeia and SPhU. For new
identification method of Hawthorn tincture preparation of test
solution,
reference solutions, system suitability solution (SST), intensity
marker, its
application, development and results were proposed. For specificity study
HPTLC-fingerprints of 13 samples of Hawthorn tinctures, which were
produced by
manufactures, were compared with HPTLC-fingerprints of laboratory
sample of
Hawthorn tincture, prepared from properly authentificated herbal raw
material
of C. laevigata (C. oxyacantha)
fructus (solvent – 70% ethanol,
ratio – 1:10) and HPTLC-fingerprints of C. laevigata
(C. oxyacantha) fructus (solvent – methanol,
ratio – 1:10). For
reproducible results the research was conducted according
to standardized procedure USP <203> High-performance thin-layer
chromatography procedure for identification of articles of botanical
origin.
Visual evaluation of chromatogram of test
solutions was conducted with respect to zone position and colour of
reference
solutions, intensity was evaluated respect to
intensity marker of reference solution. Results and Discussion. According
to the proposed identification method by HPTLC, the
fingerprints of 13
different samples of Hawthorn tincture are quite consistent respect to
zone
position, color and intensity. All analyzed samples of tinctures are
met the
requirements of proposed HPTLC-test. Comparison
of HPTLC-fingerprints of 13 samples of Hawthorn tincture with the test
solutions of laboratory samples of Hawthorn tincture and Crataegi
fructus has
shown quite similar fingerprints respect to zones position and color. This,
prove the specificity of the method. Description
of results for developed HPTLC method was given. The tolerance range in
fingerprints was specified. Despite of the storage period of Hawthorn
tincture
is 3 years, the old samples of tincture, which were manufactured in
2010 year are quite similar to new one
and would passed proposed HPTLC test. This shows the chemical stability
of
marker substances of tincture during its storage period. Other
investigation in
this area is necessary. Conclusion. The specific and reproducible HPTLC
identification method of Hawthorn tincture was developed. This could be
used as
optional/alternative method for conventional TLC. Image on chromatogram
of
Hawthorn tinctures could be used as a reference image of HPTLC method
for
Hawthorn tincture and to be included in Pharmacopoeia of Ukraine or any
other
HPTLC Atlas/Reference book. Developed HPTLC identification method of Hawthorn
tincture shown the
stability of marker group of bioactive substances (flavonoids) during
the
expiration period of tincture.
Key words: Hawthorn tincture, identification, HPTLC
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56-60
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ÑÓÄÎÂÎ-ÔÀÐÌÀÖÅÂÒÈ×ÍÅ
ÄÎÑ˲ÄÆÅÍÍß ÎÐÃÀͲÇÀÖ²¯
ÄÎÑÒÓÏÍÎÑÒ² ÎÁ²ÃÓ Ë²ÊÀÐÑÜÊÈÕ ÇÀÑÎÁ²Â ÄËß ÕÂÎÐÈÕ ÍÀ ÇËÎßʲÑͲ
ÍÎÂÎÓÒÂÎÐÅÍÍß Ó
ÑØÀ
Øàïîâàëîâ Â.Â. (ìîë.), Çáðîæåê Ñ.²., Øàïîâàëîâà Â.Î.,
Øàïîâàëîâ Â.Â., Êóëèêîâà Î.Â.
FORENSIC
AND PHARMACEUTICAL RESEARCH OF ORGANIZATION OF AVAILABILITY OF THE
MEDICINES
FOR PATIENTS WITH MALIGNANCIES IN THE UNITED STATES
Shapovalov
V.V. (Jr.), Zbrozhek S.I., Shapovalova V.A., Shapovalov V.V., Kulykova
O.V.
http://doi.org/10.5281/zenodo.167439
Introduction. The
incidence of cancer in recent years has increased significantly. It is
therefore particularly important today is the issue of provision for
patients
with malignancies with drugs. It is important to research the level of
organization of availability of the anesthetic therapy to ensure the
availability of pharmacotherapy for cancer patients worldwide. Material and
methods. For the purpose of the study analyzed legislation,
regulations of some
states in the US that provide availability of narcotic analgesic drugs
for
patients with malignant neoplasms. The paper used the following
methods:
comparative, documentary, legal, medical, pharmaceutical and graphical
analysis. Results and discussion.
Noted the increase in expenditure on
pharmaceutical provision for patients with malignancies on an
outpatient basis
in the US. During the study of the legislative and regulatory acts of
the USA
found that payments for treatment possible by insurance companies as
part of
the agreement, which in turn depend on the patient’s age, number of
family
members, their total income and so on. Coupons can be used to pay for
the cost
of medicines, but not all pharmacies accept coupons. There are
charities with
funds which are partially covered for the cost of chemotherapy and
adjuvant
therapy (analgesics, antiemetic, antipsychotics, drugs). Found, that in
New
York doctor may prescribe analgesic medication to the patient without
limitation in the frequency and dose. It is not therapeutic use of
analgesics,
their improper accounting, drug addiction patient has contraindications
to the
prescription of narcotic analgesic drugs with malignant neoplasms.
Reviewed
examples from forensic and pharmaceutical practice, pharmaceutical
violation of
the US laws that regulate the accessibility of patients with
malignancies to
narcotic analgesic drugs. So, there have been cases of fake
prescriptions for
narcotic drugs, selling drugs, the shelf life has expired. Police of
New Jersey
the fact of abduction of injectable morphine and its falsification of
employee
hospital. Conclusion.
Conducted a study of the availability of narcotic
analgesic drugs for patients with malignancies in the US from a
position of
forensic pharmacy. Made an analysis of mortality from malignancies
worldwide.
Studied pharmaceutical legislation of Nevada, Texas, Georgia, New York
in the
pharmaceutical industry of providing patients with malignancies with
narcotic
analgesic drugs.
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61-64
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