scholarly journals Immunological NBT Test – Indicator of Phagocytic and Metabolic Activity of Neutrophilic Granulocytes in Complications of Ent Organs Diseases

2018 ◽  
Vol 11 (1) ◽  
pp. 187-190
Author(s):  
Zarema N. Lovpache ◽  
Tamara G. Tlupova ◽  
Aslan A. Teuvov ◽  
Arthur M. Baziev ◽  
Diana A. Teuvazhukova
Keyword(s):  
Infectious Diseases ◽  
Metabolic Activity ◽  
Experimental Methods ◽  
Neutrophilic Granulocytes ◽  
Nbt Test

Infectious diseases associated with diseases of ENT organs are quite common. Therefore, this problem requires detailed study. The main goal of the work is to characterize the immunological NBT test – the index of phagocytic and metabolic activity of neutrophilic granulocytes in cases of complications of ENT diseases. To achieve this goal and study the problem in detail, the authors used experimental methods. The experiment was conducted on 40 patients from 32 to 65 years. The authors found that during the eruptive phase of the disease, the percentage of NBT positive granulocytes and tetrazolium activity significantly increased and amounted to 30-40%, while the control group’s data were the following – the activity of the NBT test was 4.9-6.8%. It was also found that the NBT test in patients with otitic meningitis was 26-62%, while in all patients with serous meningitis, the level of the NBT test was low – 6-11.

Mechanisms involved in effects of antimicrobial peptides, bactenectins ChBac3.4, ChBac5, and mini-ChBac7.5Nb, on bacterial cells

2017 ◽  
pp. 43-50
Author(s):  
О.В. Шамова ◽  
М.С. Жаркова ◽  
П.М. Копейкин ◽  
Д.С. Орлов ◽  
Е.А. Корнева
Keyword(s):  
Escherichia Coli ◽  
Antimicrobial Activity ◽  
Innate Immunity ◽  
Infectious Diseases ◽  
Metabolic Activity ◽  
Capra Hircus ◽  
Bacterial Cells ◽  
Antibiotic Resistant ◽  
Resistant Strains

Антимикробные пептиды (АМП) системы врожденного иммунитета - соединения, играющие важную роль в патогенезе инфекционных заболеваний, так как обладают свойством инактивировать широкий спектр патогенных бактерий, обеспечивая противомикробную защиту живых организмов. В настоящее время АМП рассматриваются как потенциальные соединения-корректоры инфекционной патологии, вызываемой антибиотикорезистентными бактериями (АБР). Цель данной работы состояла в изученим механизмов антибактериального действия трех пептидов, принадлежащих к семейству бактенецинов - ChBac3.4, ChBac5 и mini-ChBac7.5Nb. Эти химически синтезированные пептиды являются аналогами природных пролин-богатых АМП, обнаруженных в лейкоцитах домашней козы Capra hircus и проявляющих высокую антимикробную активность, в том числе и в отношении грамотрицательных АБР. Методы. Минимальные ингибирующие и минимальные бактерицидные концентрации пептидов (МИК и МБК) определяли методом серийных разведений в жидкой питательной среде с последующим высевом на плотную питательную среду. Эффекты пептидов на проницаемость цитоплазматической мембраны бактерий для хромогенного маркера исследовали с использованием генетически модифицированного штамма Escherichia coli ML35p. Действие бактенецинов на метаболическую активность бактерий изучали с применением маркера резазурина. Результаты. Показано, что все исследованные пептиды проявляют высокую антимикробную активность в отношении Escherichia coli ML35p и антибиотикоустойчивых штаммов Escherichia coli ESBL и Acinetobacter baumannii in vitro, но их действие на бактериальные клетки разное. Использован комплекс методик, позволяющих наблюдать в режиме реального времени динамику действия бактенецинов в различных концентрациях (включая их МИК и МБК) на барьерную функцию цитоплазматической мембраны и на интенсивность метаболизма бактериальных клеток, что дало возможность выявить различия в характере воздействия бактенецинов, отличающихся по структуре молекулы, на исследуемые микроорганизмы. Установлено, что действие каждого из трех исследованных бактенецинов в бактерицидных концентрациях отличается по эффективности нарушения целостности бактериальных мембран и в скорости подавления метаболизма клеток. Заключение. Полученная информация дополнит существующие фундаментальные представления о механизмах действия пролин-богатых пептидов врожденного иммунитета, а также послужит основой для биотехнологических исследований, направленных на разработку на базе этих соединений новых антибиотических препаратов для коррекции инфекционных заболеваний, вызываемых АБР и являющимися причинами тяжелых внутрибольничных инфекций. Antimicrobial peptides (AMPs) of the innate immunity are compounds that play an important role in pathogenesis of infectious diseases due to their ability to inactivate a broad array of pathogenic bacteria, thereby providing anti-microbial host defense. AMPs are currently considered promising compounds for treatment of infectious diseases caused by antibiotic-resistant bacteria. The aim of this study was to investigate molecular mechanisms of the antibacterial action of three peptides from the bactenecin family, ChBac3.4, ChBac5, and mini-ChBac7.5Nb. These chemically synthesized peptides are analogues of natural proline-rich AMPs previously discovered by the authors of the present study in leukocytes of the domestic goat, Capra hircus. These peptides exhibit a high antimicrobial activity, in particular, against antibiotic-resistant gram-negative bacteria. Methods. Minimum inhibitory and minimum bactericidal concentrations of the peptides (MIC and MBC) were determined using the broth microdilution assay followed by subculturing on agar plates. Effects of the AMPs on bacterial cytoplasmic membrane permeability for a chromogenic marker were explored using a genetically modified strain, Escherichia coli ML35p. The effect of bactenecins on bacterial metabolic activity was studied using a resazurin marker. Results. All the studied peptides showed a high in vitro antimicrobial activity against Escherichia coli ML35p and antibiotic-resistant strains, Escherichia coli ESBL and Acinetobacter baumannii, but differed in features of their action on bacterial cells. The used combination of techniques allowed the real-time monitoring of effects of bactenecin at different concentrations (including their MIC and MBC) on the cell membrane barrier function and metabolic activity of bacteria. The differences in effects of these three structurally different bactenecins on the studied microorganisms implied that these peptides at bactericidal concentrations differed in their capability for disintegrating bacterial cell membranes and rate of inhibiting bacterial metabolism. Conclusion. The obtained information will supplement the existing basic concepts on mechanisms involved in effects of proline-rich peptides of the innate immunity. This information will also stimulate biotechnological research aimed at development of new antibiotics for treatment of infectious diseases, such as severe in-hospital infections, caused by antibiotic-resistant strains.

scholarly journals PECULIARITIES OF THE WOUND PROCESS DEVELOPMENT IN PATIENTS WITH THERMAL INJURY WHEN USED SILVER PREPARATIONS IN BURN WOUNDS SURGICAL TREATMENT

2021 ◽  
pp. 81-87
Author(s):  
N. V. Tuzyuk
Keyword(s):  
Metabolic Activity ◽  
Functional Activity ◽  
Thermal Injury ◽  
Thermal Damage ◽  
Main Group ◽  
The Body ◽  
Metabolic Reaction ◽  
Neutrophilic Granulocytes ◽  
Burn Wound ◽  
Significant Activation

Fundamental research in the field of burn wound development process highlights new pathogenetic mechanisms that underlie the improvement of medical practice, the main purpose of which is to create optimal conditions for early regeneration of the affected skin, as well as to eliminate the development of local and general complications. Irrational general and local treatment of wounds contribute to the slowing down of regenerative processes, thereby worsening the prognosis and consequences of thermal injury. It was found out that in patients of both study groups in the first period of the study the characteristics of the cellular composition of the wound impressions of the wound bottom of the area of thermal damage correspond to the degenerative type. In patients of the main group in the second term of the study in the area of thermal damage cells with high levels of functional and metabolic activity begin to appear, which increase antimicrobial resistance and reduce the risk of generalization of the infectious process which is the main source of burns. In patients of the main group, an increase in the activity of neutrophilic granulocytes in the induced test was found in the second term of the study, which, in our opinion, indicates the involvement of neutrophilic granulocytes with high functional and metabolic activity. Under conditions of significant activation of resident monocytes in response to injury in patients of the main group in the second and third term, the area of thermal damage migrates functionally effective monocytes. In patients of the main group the preservation of the activity of myeloperoxidase - the main enzyme of the oxygen-dependent phase of phagocytosis at the minimum subcompensated level, which leads to the preservation of the oxygen-dependent metabolic reaction of neutrophilic granulocytes in the subsequent study, Increases in the content of PAS-positive substances in neutrophilic granulocytes indicate the preservation of the functional reserve for activation and completion of the oxygen-independent phase of phagocytosis. The use of the developed treatment promotes the involvement of functionally active phagocytic cells in the burn wound, reduces the toxic load on monocytes and neutrophils, which helps to maintain their functional activity at the subcompensated level. These trends in patients of the main group reduce the risk of both local and general infectious complications of burn disease. It was found that the level of spontaneous functional activity in the HCT test is associated with the activity of resident monocytes. They form the primary barrier that protects the body from infection or harmful macromolecular complexes. Monocytes - "inflammatory", which constantly come from the free pool and their functional activity against microbial antigens is much higher than resident monocytes. An indicator of the activity of monocytes - "inflammatory" is an indicator of functional activity in the induced HCT test. The results of the study indicate that under conditions of significant activation of resident monocytes in response to injury in patients of the main group in the second and third terms of the area of thermal damage migrate functionally effective monocytes.

Metabolic activity of neutrophilic granulocytes measured with chemiluminescence test (cl) in patients with allergic hypersensitivity to food

2012 ◽  
Vol 26 (2) ◽  
Author(s):  
Magdalen Żbikowska-Gotz ◽  
Krzysztof Pałgan ◽  
Ewa Socha ◽  
Michał Przybyszewski ◽  
Andrzej Kuźmiński ◽  
...  
Keyword(s):  
Metabolic Activity ◽  
Neutrophilic Granulocytes

scholarly journals Phagocytic and metabolic activity of neutrophils of rats in the early stages of post-embryonic development under the influence of biologically active substances of the salt extract of Hirudo verbana

2016 ◽  
Vol 7 (2) ◽  
pp. 96-100 ◽  
Author(s):  
R. F. Aminov ◽  
A. K. Frolov
Keyword(s):  
Metabolic Activity ◽  
The Body ◽  
Biologically Active ◽  
Control Group ◽  
Standard Diet ◽  
Immunological Parameters ◽  
Ether Anesthesia ◽  
Saline Extract ◽  
Nbt Test ◽  
Salt Extract

The nitroblue tetrazolium test reflects the degree of activation of oxygen-dependent mechanisms of bactericidal activity of phagocytotic cells, so the increase in nbt-test and phagocytic indices shows increased activation in the bacterial process and enhanced protection with the help of the absorption ability of the cells in the body. We examined females of nonlinear rats after feeding of their offspring, which 2 weeks before and 2 weeks after mating were injected intraperitoneally with antigens of saline extract of medical leeches (4 injections) and the offspring at days 1, 15, 30, 45, 60. The dosage of salt extract antigens was carried out by determining the protein concentrations using the Lowry protein assay. The animals were immobilized using an immobilizing device. The animals were distributed into three groups: the first research and experimental group of animals was subject to the influence of medical leech salt extract antigens to the amount of 0.5 ml (at the rate of 3 µg/g of animal weight); the second group of intact animals was not subject to intervention, the third control group of animals was administered intraperitoneally saline 0.5 ml of 9% saline solution. We studied females after feeding offspring and offspring at 1, 15, 30, 60 days. The animals were kept in vivarium conditions in individual cages on a standard diet. In the experiment we used 60 females of non-linear rats and 300 of their offspring. All animals were decapitated under ether anesthesia. After that, we took blood, made a solution with crystal heparin and examined the laboratory parameters: total number of leukocytes and leukocyte formula. Evaluation of nonspecific resistance of the organism was investigated using tests which characterize the activity of neutrophils (nbt-test, phagocytosis indices). The results of our studies revealed a positive effect of antigens of saline extract of medical leeches on the absorptive and metabolic activity of neutrophils in the females and their offspring in all study periods compared with the control, an increased number of leukocytes, due to the change of the process of differentiation and deposition of cells in immunogenic organs on proliferation. Changes in immunological parameters indicate the immunostimulatory effect of AG ML on leukopoiesis and on the functional activity of neutrophils – the first line of defense against entry into the body of a variety of bacteria, fungi and protozoa. 

scholarly journals Study of innate factors in the local immune defense of the genital organs of dogs and cats

2019 ◽  
Vol 21 (93) ◽  
pp. 98-102
Author(s):  
M. M. Zhelavskyi
Keyword(s):  
Epithelial Cells ◽  
Immune Defense ◽  
Sexual Cycle ◽  
Immune Homeostasis ◽  
Neutrophilic Granulocytes ◽  
Cytoplasmic Inclusions ◽  
Extracellular Traps ◽  
Nbt Test ◽  
Mucous Membranes ◽  
Antimicrobial Protection

Immune system of reproductive organs of animals is represented by a number of cellular and humoral protection factors. Recently, the attention of scientists was attracted by the role of immunocompetent cells, which integrally provide immune homeostasis at all stages of the reproductive function. The purpose of the work was to study the cellular factors of immune defense of mucous membranes of genital organs of dogs and cats. The cytological composition and determination of immunological parameters of vaginal microscope slide were determined by patented methods. Local immunity was determined by the parameters of extracellular protection mechanisms, which consisted in the study of the Oxygen-dependent potential of neutrophil granulocytes (NBT-test) and their ability to form extracellular traps (NETs). Our studies have found that in different stages of the sexual cycle in beach and cats, along with cytological changes in epithelial cells, changes occur in phagocytic protection. In particular, in the post-infusion period there was an increase in the number of intermediate and basal epithelial cells on the surface of which were adhered microorganisms. Along with epithelial cells, activated phagocytes were grouped. Neutrophils showed cytochemical reactivity in the NBT-test, and also formed NETs. At the same time, activated phagocytes absorbed microorganisms, and some epithelial cells formed specific cytoplasmic inclusions, which obviously also has an important role in the antimicrobial immunity of the mucosa. Some neutrophilic granulocytes were in a state of apoptosis. During the sexual cycle in the organism of dogs and cats there are morphology and functional changes in the genital organs, which was accompanied by a change in the cytological picture of vaginal microscope slides. It was established that the antimicrobial protection system plays an important role in the neutrophilic granulocytes, the functional activity of which also has a change. The main mechanisms of extracellular antimicrobial protection of microphages are realized with the involvement of Oxygen species in the excretion system of active forms and the formation of neutrophilic extracellular traps. Epithelial cells are also involved in the formation of immune homeostasis of genital mucous membranes.

scholarly journals FUNCTIONAL PROPERTIES OF BLOOD NEUTROPHILS AND PERITONEAL EXUDATE IN PATIENTS WITH PERITONITIS

2019 ◽  
pp. 22-29
Author(s):  
A. Stasenko
Keyword(s):  
Functional Properties ◽  
Metabolic Activity ◽  
Abdominal Cavity ◽  
The Body ◽  
Peritoneal Exudate ◽  
Innate Defense ◽  
Local Protection ◽  
Cellular Factors ◽  
Nbt Test ◽  
Blood Neutrophils

The aim of the study is to investigate the functional properties of neutrophils in the blood and peritoneal exudate in patients with peritonitis. Materials and methods. The metabolic activity of neutrophils determined in patients with peritonitis and in patients of the control groups in the blood, peritoneal exudate and effusion of the abdominal wall on the 1–2 day of the postoperative period in the Sp and St test with nitro blue tetrazolium (NBT), as well as reserve indicator (RI) of metabolic activity. We studied the percentage of active neutrophils in the phagocytosis –PhI ( index) and the number of absorbed Candida albicans particles on average by one neutrophil –PhN(number). Results and their discussion.The peculiarities of the cellular factors of local protection decreased metabolic activity of neutrophils in the abdominal cavity in the Sp NBT test compared to blood neutrophils and an increased reserve during stimulation. For the comparisons which included the activity of neutrophils abdominal contents in patients with peritonitis versus in control established that neutrophils in the abdominal cavity with peritonitis have a reduced intensity of phagocytosis. Patients with pancreatic peritonitis have inverse correlations between the amount of NBT-positive blood NG in the SP NBT test and IR of blood NG and between the amount of NBT-positive blood NG in the Sp NST test and PhI of blood NG, while patients with abdominal peritonitis do not have correlations between indicators of innate defense of the body. Conclusions. Peritonitis was associated with functional disorders of NG. The leading areas of disorders are significant activation of metabolic processes of NG and inhibition of the phagocytic function of NG blood and peritoneal exudate. Patients with pancreatic peritonitis had inverse correlations between the indicators of the innate defense of the body, while patients with abdominal peritonitis did not have correlations.

The nature of metabolic and functional state of the innate antimicrobial protection factors in persons with rhinosinusitis

2021 ◽  
pp. 70-75
Author(s):  
N. V. Kornova
Keyword(s):  
Relative Number ◽  
Nasal Secretion ◽  
Quantitative Composition ◽  
Neutrophilic Granulocytes ◽  
Test Results ◽  
Lysosomal Activity ◽  
Cellular Factors ◽  
Nbt Test ◽  
Antimicrobial Protection ◽  
Biocidal Properties

The purpose is to study the features of metabolic and functional activity of cellular factors of innate immunity in nasal secretion in persons with rhinosinusitis. Material and methods. We studied the qualitative and quantitative composition of leukocytes of the nasal secretion of patients with rhinosinusitis, their viability, phagocytic and lysosomal activity, oxygen-dependent metabolism of neutrophilic granulocytes in the NBT test. Results. It was demonstrated that in patients with rhinosinusitis, an increase in the absolute and relative number of viable neutrophilic granulocytes, an increase in their lysosomal activity, a decrease in the activity and intensity of phagocytosis, and inhibition of biocidal properties according to the spontaneous NBT test are recorded. Conclusion. The revealed increase in the number of viable neutrophilic granulocytes, an increase in their lysosomal activity, a traced decrease in the activity and intensity of phagocytosis with inhibition of biocidal properties confirm the need to search for additional methods of rhinosinusitis therapy.

scholarly journals Metabolic activity of neutrophilic granulocytes and possible ways of its correction in patients with acute coronary syndrome

2011 ◽  
Vol 57 (2) ◽  
pp. 35-42 ◽  
Author(s):  
NA Ryzhkova ◽  
◽  
TI Havrylenko ◽  
OM Parkhomenko ◽  
SM Kozhukhov ◽  
...  
Keyword(s):  
Acute Coronary Syndrome ◽  
Metabolic Activity ◽  
Neutrophilic Granulocytes ◽  
Coronary Syndrome
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