Phagocytic Leukocytes Research Articles

Determination of phagocytic activity of S. Aureus strains isolated from children with measles

Summary. S. aureus is one of the world's most common pathogens of infectious diseases. It is known that the development of infection is facilitated by various factors and factors of virulence of staphylococci. Much less is known about their interaction with each other and the protective mechanisms of the host's immune system. The importance of immunocompetent cells in the development of the inflammatory process was the basis for studying the ability to phagocytose strains of Staphylococcus aureus isolated from the nasopharynx of children with measles. The aim of the study – to determine the ability to phagocytose Staphylococcus aureus strains isolated from children with measles by neutrophilic granulocytes and peripheral blood monocytes. Materials and Methods. The phagocytic response was assessed using the following indicators: phagocytosis activity or the percentage of phagocytic-active leukocytes, phagocytosis index – the average number of absorbed microbes per phagocyte. During the study, two groups were formed: group 1 – experimental (35 patients with measles); group 2 – control (samples of normal microflora from almost healthy people – 15 persons). The material from the oropharynx was taken with a sterile swab, examined by bacteriological method. The quantitative composition of bacteria was reflected in the number of colony forming units (CFU) in terms of dilution of the test material. Identification of microorganisms was performed according to standard schemes. Phagocytosis studies were performed by the method of Kost and Stenko. Results. During the study, it was found that neutrophilic granulocytes play an important role in the process of phagocytosis. Phagocytic activity of neutrophils isolated from children with measles and monocytes was less than 46.9 % and 38.8 % compared with the group of healthy individuals. There was also a decrease in the phagocytic index in phagocytic leukocytes relative to the culture of S. aureus isolated from children with measles: neutrophils – 31.7 %, monocytes – 38.6 %. Conclusions. The decrease in the phagocytic ability of Staphylococcus aureus culture isolated from children with measles by phagocytic cells in the experimental group is apparently due to the presence of virulence factors of the pathogen, which significantly inhibit phagocytosis and cause the ability of microorganisms to persist for a long time

Lymphocyte Proliferation and Nitric Oxide-Producing Activities of Lupeol Isolated From Red Dragon Fruit (Hylocereus polyrhizus) Extract

Background: Hylocereus polyrhizus has activities as antimicrobial agent, anti-hypercholesterolemia, anti-diabetic (diabetes mellitus), cardiovascular risk reduction, health supplement, and melanoma cell suppression. The extracts from the peels of H. polyrhizus were able to increase phagocytic ability, cell numbers and leukocytes and to influence relative spleen weights in the formation of body immune system in male rats. The fruit peels contained phenolics, flavonoids, carotenoids, and anthocyanins. This study investigated the active compounds of H. polyrhizus peels, which are able to increase immune system of human body.Materials and method: In vitro assay was applied to examine the active compounds, identified as lupeol, obtained from isolated extract of red dragon fruit for their lymphocyte proliferation and nitric oxide (NO)-producing activities. Lymphocyte proliferation assay was performed with 3-4.5-dimethylthiazol-2-yl)-2.5-diphenyltetrazolium bromide (MTT) method. The cell control was lymphocyte cell suspension in RPMI medium added with phytohaemaglutinine (PHA). The NO measurement was conducted with nitric solvent and Greiss reagent.Results: The ANOVA analysis of the average optical density (OD) of lymphocyte proliferation showed that the addition of isolated lupeol at the concentrations of 6.25, 12.5, 25, 50 and 100 µg/mL were able to improve lymphocyte proliferation and activate the NO production in the rats with treatment of positive control. Conclusion: Isolated lupeol at concentrations of 6.25, 12.5, 25, 50 and 100 µg/mL revealed significant difference with medium control and cell control. It was able to increase effects on lymphocyte proliferation and NO production. Therefore, the lupeol which was isolated might have high potential to be an immunostimulant.Keywords: Hylocereus polyrhizus, lupeol, lymphocyte proliferation, nitric oxide production

Growth Performance, Cellular Immunity and Blood Chemical Analysis of Two Broilers Strains after Phytogenic Supplementation

Abstract: Genetically, the basis of all commercial breeds is the same. Therefore, the selection pressure for traits such as performance and carcass yield results in particular products. Genetic variation of two different broiler strains (Avian48 and Cobb500) in response to black pepper and red hot pepper supplementation was studied. Two hundred chicks from each broiler strain were randomly distributed on the following treatments, 1. Control treatment; 2. Red hot pepper treatment (0.5 %); 3. Black pepper treatment (0.5%); 4. Mixture treatment (0.25 % black pepper plus 0.25 % red pepper), these additives were added to drinking water from the first week until the fifth week of age, every group containing 50 birds distributed on five replicates (10 birds/replicate). The growth performance, carcass traits, abdominal fat, edible giblets, blood pictures, cellular immunity, and chemical blood lipid profile were estimated. Cobb500 broilers strain respond better to water supplement of red hot pepper, black pepper, and their mixture than Avian48 strain. Moreover, adding red hot pepper (0.5), black pepper (0.5), and their mixture (0.25+0.25) to drinking water significantly improved growth performance compared to control treatments with the best results obtained from mixture supplementation. However, it's better to supplying these additives until the third week of age, especially black pepper. Using black pepper, red hot pepper, and their mixture as feed additives by 0.5% had a significant role in further improving carcass quality by lowering fat percent and blood cholesterol. LDL so decrease their percent in meat and increasing the dressing and carcass cuts as a general, with the best results reported for mixture supplement. Additionally, these herbs had the desired effect on improving broilers' cellular immunity by increasing phagocytic index and differential leukocyte count. Finally, further examinations are required to studying the response of other broiler strains and breeds to these supplements and evaluating its effect on carcass characteristics, abdominal fat, cellular immunity, blood picture, and lipid profile if used within the first three weeks of age only.

The role of phagocytic leukocytes following flexible ureterenoscopy, for the treatment of kidney stones: an observational, clinical pilots-study

BackgroundThe number of patients undergoing flexible ureterenoscopy (FURS) for the treatment of kidney stones (renal calculi) is increasing annually, and as such the development of post-operative complications, such as acute kidney injury (AKI), haematuria and infection is likely to increase. Phagocytic leukocytes are white blood cells that help fight foreign material such as bacteria and viruses, and they are intrinsically involved in the inflammatory reaction. Investigating the role of phagocytic leukocytes following FURS has not been widely researched. The main aim of the study was to evaluate the role phagocytic leukocytes (neutrophils and monocytes) function, in patients undergoing FURS for the treatment of kidney stones (renal calculi).MethodsFourteen consecutive patients aged between 27 and 70 years (median 49.5 years) undergoing FURS for the treatment of kidney stones were recruited (seven males, seven females). Blood samples were collected from each patient at four time points: baseline (pre-operatively) followed by 30, 120 and 240 min post-operatively. Mononuclear (MN) and polymorphonuclear (PMN) leukocyte sub-populations were isolated by density gradient centrifugation techniques. Neutrophil and monocyte cell function was investigated by measuring the cell surface expression of CD62L (L-selectin), CD11b (Mac-1), CD99 and the intracellular production of hydrogen peroxide (H2O2), via flow cytometry.ResultsSignificant increases was observed in monocyte CD62L expression post FURS for the treatment of kidney stones (p ≤ 0.05); while significant decreases were observed in neutrophil CD62L. The levels of the other activation markers CD11b, CD99 and H2O2 corresponded to the increases and decreases seen in CD62L for monocytes and neutrophils respectively, though the changes were not statistically significant (p > 0.05). Limiting factors for this study were the relatively small sample size, and restriction on the recruitment time points.ConclusionsThis study demonstrates that following FURS for the treatment of kidney stones, monocytes are rapidly activated and produce potent reactive oxygen intermediates. Interestingly, the pattern of expression in neutrophils suggests that these cells are deactivated in response to the treatment. The leukocyte biomarkers assessed during this investigation may have a role in monitoring the ‘normal’ post-operative response, as no complications occurred in any of the patients; or may help predict potential infectious complications (e.g. urosepsis) that can occur during the post-operative period. This data, however, will need to be validated and reproduced in larger multi-centre studies.

Phagocytic Activity of Leukocytes in Harp Seals.

Phagocytic and cytochemical parameters of peripheral blood leukocytes in adult and 1.5-month-old harp seal pups were studied. Phagocytic number, proportion of phagocytic leukocytes, and phagocytic activity, average number of absorbed latex particles per leukocyte, were determined. Differences between groups of seals by average values of these indicators have not been revealed. A correlation analysis revealed a significant relationship between the phagocytic activity of leukocytes and their glycogen content in adult seals (r = 0.89). Content of cationic bactericidal protein (CP) in leukocytes of adult animals was 10-12 times higher than in pups. Correlation of CP and eosinophil count in adults was high (r = 0.88). Adult had significantly more eosinophils than pups. The noted features can provide higher efficiency of phagocytosis and destruction of absorbed bacteria in adult seals as compared to pups, despite the high level of indicators of the first, absorbing, phase of phagocytosis in the latter.

Milk fat globule epidermal growth factor 8 (MFG-E8) on monocytes is a novel biomarker of disease activity in systemic lupus erythematosus.

Milk fat globule epidermal growth factor (MFG-E8) is related secreted protein which links phosphatidylserine on apoptotic cells and integrin αvβ3/5 on phagocytes. To clarify the clinical significance of MFG-E8 in SLE, we analyzed the correlation between expression level of MFG-E8 in circulating phagocytic leukocytes and clinical parameters of patients. The study was conducted under a multi-center, prospective cohort design. Patients with one or both BILAG A or B, or SLEDAI- 2 K ≥ 4 with clinical symptoms were defined as the active SLE group. Expression of MFG-E8 on monocytes and concentration in serum were measured by FACS and ELISA, respectively. 96 subjects were enrolled. The absolute number and proportion of MFG-E8-positive monocytes to total monocytes were significantly higher in the active SLE group (p < 0.01). Importantly, the proportion was also significantly correlated with SLEDAI-2K, clinical SLEDAI, as well as serum levels of anti-ds-DNA antibody and complement and C1q. In addition, the proportion of MFG-E8-positive monocytes to total monocytes was significantly decreased from baseline in active SLE patients after 6 months' treatment and increased concordantly with disease activity in 6 refractory cases. Further, in receiver operating characteristic curve analysis for discrimination between active and inactive SLE, the AUC of the proportion of MFG-E8 was 0.854, which was equivalent to classical activity markers such as anti-ds DNA antibody (0.776), complement (0.897) and C1q (0.815). The proportion of MFG-E8-positive monocytes to total monocytes in peripheral blood was positively associated with disease activity in SLE and may be a novel biomarker of disease activity.

Attenuated P. falciparum Parasite Shows Cytokine Variations in Humanized Mice.

A recently developed humanized mouse has been used to assess the immune response evoked against the isolated attenuated C9 parasite clone (C9-M; carrying a single insertion disrupting the open reading frame (ORF) of PF3D7_1305500) of Plasmodium falciparum. Significant human RBC engraftment was achieved by ameliorating the residual non-adaptive immune response using clodronate-loaded liposome treatment. Controlled reactive professional phagocytic leukocytes in immunodeficient mice allowed for sizeable human blood chimerism and injected huRBCs acted as bona fide host cells for P. falciparum. huRBC-reconstituted immunodeficient mice received infectious challenge with attenuated P. falciparum C9 parasite mutants (C9-M), complemented (C9-C), and wild type (NF54) progenitors to study the role of immune effectors in the clearance of the parasite from mouse circulation. C9-M and NF54 parasites grew and developed in the huRBC-reconstituted humanized NSG mice. Further, the presence of mutant parasites in deep-seated tissues suggests the escape of parasites from the host's immune responses and thus extended the survival of the parasite. Our results suggest an evasion mechanism that may have been employed by the parasite to survive the mouse's residual non-adaptive immune responses. Collectively, our data suggest that huRBCs reconstituted NSG mice infected with attenuated P. falciparum is a valuable tool to explore the role of C9 mutation in the growth and survival of parasite mutants and their response to the host's immune responses. This mouse might help in identifying novel chemotherapeutic targets to develop new anti-malarial drugs.

Is Antioxidant Therapy a Useful Complementary Measure for Covid-19 Treatment? An Algorithm for Its Application.

Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) causes the corona virus disease-19 which is accompanied by severe pneumonia, pulmonary alveolar collapses and which stops oxygen exchange. Viral transmissibility and pathogenesis depend on recognition by a receptor in the host, protease cleavage of the host membrane and fusion. SARS-CoV-2 binds to the angiotensin converting enzyme 2 receptor. Here, we discuss the general characteristics of the virus, its mechanism of action and the way in which the mechanism correlates with the comorbidities that increase the death rate. We also discuss the currently proposed therapeutic measures and propose the use of antioxidant drugs to help patients infected with the SARS-CoV-2. Oxidizing agents come from phagocytic leukocytes such as neutrophils, monocytes, macrophages and eosinophils that invade tissue. Free radicals promote cytotoxicity thus injuring cells. They also trigger the mechanism of inflammation by mediating the activation of NFkB and inducing the transcription of cytokine production genes. Release of cytokines enhances the inflammatory response. Oxidative stress is elevated during critical illnesses and contributes to organ failure. In corona virus disease-19 there is an intense inflammatory response known as a cytokine storm that could be mediated by oxidative stress. Although antioxidant therapy has not been tested in corona virus disease-19, the consequences of antioxidant therapy in sepsis, acute respiratory distress syndrome and acute lung injury are known. It improves oxygenation rates, glutathione levels and strengthens the immune response. It reduces mechanical ventilation time, the length of stay in the intensive care unit, multiple organ dysfunctions and the length of stay in the hospital and mortality rates in acute lung injury/acute respiratory distress syndrome and could thus help patients with corona virus disease-19.

Genetic Characteristics, Infectious, and Noninfectious Manifestations of 32 Patients with Chronic Granulomatous Disease

Background: Chronic granulomatous disease (CGD) is a rare genetic disorder characterized by failure of phagocytic leukocytes to destroy certain microbes. We present a study on CGD patients enrolled at a single medical center concerning the infectious and noninfectious complications and genetic properties of the disease. Methods: Icotinamide adenine dinucleotide phosphate oxidase activity and the expression of flavocytochrome b₅₅₈ were measured by flow cytometry, and clinical outcomes of the patients were listed in relation to the genetic results. Results: The clinical and genetic findings of 32 pediatric cases with CGD from 23 families were enrolled. Pneumonia and anemia were the most common infectious and noninfectious symptoms. Genetic analysis showed that 10 families (43.5%) carried CYBB variants and 13 families (56.5%) have autosomal recessive (AR) CGD, in which 6 families (26%) carried NCF1 variants, 4 (17.4%) carried CYBA variants, and 3 (13%) carried NCF2 variants. The median age of clinical onset was 3.3 and 48 months for patients with X-linked CGD (X-CGD) and AR-CGD, respectively. The onset of symptoms before age 1 year was 94% in X-CGD, 28.5% in AR-CGD, and 12.5% in patients with oxidase residual activity. Moreover, a de novo germline mutation at c.1415delG in CYBB (OMIM#300481) and a novel c.251_263del13bp in CYBA (OMIM#608508) were also investigated. Conclusions: Ihydrorhodamine-1,2,3 assay could not detect carrier mother in de novo case with CYBB variant. Most X-CGD patients have the onset of symptoms before age 1 year. Additionally, residual oxidase activity in AR-CGD causes a delay in onset, diagnosis, and prophylaxis. The protective role of residual activity is limited while the infection is ongoing and becoming serious.

Reactions of Subcutaneous Cellular Tissue in Response to the Implantation of a Polypropylene Endoprosthesis Modified with a Polycaprolactone Solution by Electrospinning

Objective: to study the morphological features of subcutaneous tissue in response to the implantation of a polypropylene mesh endoprosthesis (PSE) modified with a polycaprolactone solution by the method of electrospinning. Material and methods. The object of the study was modified polypropylene mesh endoprostheses implanted in experimental animals. The animals were removed from the experiment on the 3rd, 7th, 60th day of the experiment. Subcutaneous fatty tissue along with the implanted material and regional lymph nodes were fixed in a 10 % solution of neutral formalin, followed by histological posting according to the standard technique and filled with paraffin. Histological sections 3-4 μm thick were stained with hematoxylin and eosin. The morphological assessment of reactions to the implantation of the materials was carried out by means of the calculation of the cellular elements of inflammatory and connective tissue origin along the periphery of the implants (neutrophilic leukocytes, phagocytic neutrophilic leukocytes, degenerating neutrophilic leukocytes, eosinophilic leukocytes, lymphocytes, plasmacytes, histiocytes, macrophages, fibrocytes, fibroblasts, multinucleated giant cells).Results. The study of the cellular composition has showed a statistically significant increase in the content of fibrocytes (p = 0.002) and fibroblasts (p &lt; 0.001) on the 7th day of observation around the modified PSE. On the 60th day, the number of neutrophilic leukocytes at the periphery of the modified PSE was lower than that of the control sample (p = 0.004). The enhancement of the fibroplastic properties of the samples modified with polycaprolactone by electrospinning was characterized by a less pronounced inflammatory reaction of the peripheral tissues.Conclusion. The obtained data are indicative of the fact that the modification of PSE with polycaprolactone reduces the reactive properties of endoprostheses and make it possible to recommend to introduce this method of PSE modification in clinical practice.

The Role of Formyl Peptide Receptor 1 Gene Polymorphisms in Human Colorectal Cancer

Formyl peptide receptor 1 (FPR1) belongs to G protein-coupled receptors expressed mainly in phagocytic leukocytes. The gene encoding FPR1 is highly polymorphic and related to inflammation. In this study, we investigated the single nucleotide polymorphisms (SNPs) of Fpr1 in human colorectal cancer (CRC), and analyzed the association of Fpr1 SNPs with clinicopathological parameters and some specific diagnostic markers of CRC. Although the allele and genotype frequencies of Fpr1 SNPs in CRC tissues were not significantly different from that in whole blood cells derived from healthy Chinese subjects. Significant associations were observed between genotypes of c.289C>A and distant metastasis (P=0.001), and between genotypes of c.306T>C and tumor size (P=0.016). Genotypes of c.546C>A was closer to tumor size and lymphatic invasion (P=0.012 and P=0.043, respectively). Meanwhile, genotypes of c.1037C>A was related with tumor location and differentiation (P=0.000 and P=0.005, respectively). Besides, genotypes of c.576T>C>G was related with pathological type (P=0.000). Furthermore, several Fpr1 SNP positions including c.289 (C>A) and c.576 (G>C>T) were related to the expression of P53 (P=0.004 and P=0.008, respectively), and similar results were observed between other Fpr1 SNP positions and CEA, HER2 and Ki-67 (P<0.05). Our data demonstrate that Fpr1 SNPs may play the important role in the progression and metastasis of CRC.

The N-formyl peptide receptors: contemporary roles in neuronal function and dysfunction.

N-formyl peptide receptors (FPRs) were first identified upon phagocytic leukocytes, but more than four decades of research has unearthed a plethora of non-myeloid roles for this receptor family. FPRs are expressed within neuronal tissues and markedly in the central nervous system, where FPR interactions with endogenous ligands have been implicated in the pathophysiology of several neurodegenerative diseases including Alzheimer’s disease and Parkinson’s disease, as well as neurological cancers such as neuroblastoma. Whilst the homeostatic function of FPRs in the nervous system is currently undefined, a variety of novel physiological roles for this receptor family in the neuronal context have been posited in both human and animal settings. Rapid developments in recent years have implicated FPRs in the process of neurogenesis and neuronal differentiation which, upon greater characterisation, could represent a novel pharmacological target for neuronal regeneration therapies that may be used in the treatment of brain/spinal cord injury, stroke and neurodegeneration. This review aims to summarize the recent progress made to determine the physiological role of FPRs in a neuronal setting, and to put forward a case for FPRs as a novel pharmacological target for conditions of the nervous system, and for their potential to open the door to novel neuronal regeneration therapies.

Anti-interleukin-17A and anti-interleukin-23 antibodies may be effective against Alzheimer's disease: Role of neutrophils in the pathogenesis.

IntroductionDespite the remarkable progress achieved in the research on Alzheimer's disease (AD), its exact pathogenesis is not fully understood and effective therapies do not currently exist. In order to find effective therapy for AD, I ranged extensively over the literature and found an important paper by Tiffany and colleagues.Results and ConclusionNeuroinflammation has been proposed as a possible cause or driving force of AD. The discovery by Tiffany et al. that amyloid β (Aβ) is a formylpeptide receptor 2 agonist indicated that Aβ is a potent chemoattractant for phagocytic leukocytes. Therefore, in all likelihood Aβ attracts peripheral blood neutrophils, monocytes, as well as microglia cells in brain parenchyma, and activates them. However, the role of microglia cells and their precursor monocytes in AD pathogenesis remains elusive. Recently, neutrophils were found to be present in areas with Aβ deposits in AD brain and in transgenic AD model mice. Because brain is vulnerable to the effects of reactive oxygen species (ROS) and neutrophils secrete a large amount of ROS, neutrophils look like a driving force of AD. Therefore, a possibility arises that anti‐IL‐17A and anti‐IL‐23 antibodies are effective against AD, because these antibodies can be thought to interfere with neutrophil trafficking from the bone marrow to the blood circulation and thus inhibit neutrophil infiltration into AD brain. Clinical studies using anti‐IL‐17A and anti‐IL‐23 antibodies in patients with AD are required.

SPECT imaging of lung ischemia-reperfusion injury using [99mTc]cFLFLF for molecular targeting of formyl peptide receptor 1.

Primary graft dysfunction after lung transplantation, a consequence of ischemia-reperfusion injury (IRI), is a major cause of morbidity and mortality. IRI involves acute inflammation and innate immune cell activation, leading to rapid infiltration of neutrophils. Formyl peptide receptor 1 (FPR1) expressed by phagocytic leukocytes plays an important role in neutrophil function. The cell surface expression of FPR1 is rapidly and robustly upregulated on neutrophils in response to inflammatory stimuli. Thus, we hypothesized that use of [99mTc]cFLFLF, a selective FPR1 peptide ligand, would permit in vivo neutrophil labeling and noninvasive imaging of IRI using single-photon emission computed tomography (SPECT). A murine model of left lung IRI was utilized. Lung function, neutrophil infiltration, and SPECT imaging were assessed after 1 h of ischemia and 2, 12, or 24 h of reperfusion. [99mTc]cFLFLF was injected 2 h before SPECT. Signal intensity by SPECT and total probe uptake by gamma counts were 3.9- and 2.3-fold higher, respectively, in left lungs after ischemia and 2 h of reperfusion versus sham. These values significantly decreased with longer reperfusion times, correlating with resolution of IRI as shown by improved lung function and decreased neutrophil infiltration. SPECT results were confirmed using Cy7-cFLFLF-based fluorescence imaging of lungs. Immunofluorescence microscopy confirmed cFLFLF binding primarily to activated neutrophils. These results demonstrate that [99mTc]cFLFLF SPECT enables noninvasive detection of lung IRI and permits monitoring of resolution of injury over time. Clinical application of [99mTc]cFLFLF SPECT may permit diagnosis of lung IRI for timely intervention to improve outcomes after transplantation.

The Evidence for Allogeneic Hematopoietic Stem Cell Transplantation for Congenital Neutrophil Disorders: A Comprehensive Review by the Inborn Errors Working Party Group of the EBMT.

Congenital disorders of the immune system affecting maturation and/or function of phagocytic leucocytes can result in severe infectious and inflammatory complications with high mortality and morbidity. Further complications include progression to MDS/AML in some cases. Allogeneic stem cell transplantation is the only curative treatment for most patients with these diseases. In this review, we provide a detailed update on indications and outcomes of alloHSCT for congenital neutrophil disorders, based on data from the available literature.

Inhibition of Human Neutrophil Extracellular Trap (NET) Production by Propofol and Lipid Emulsion.

Uncontrolled bacteremia is a common and life threatening condition that can lead to sepsis and septic shock with significant morbidity and mortality. Neutrophil granulocytes, the most abundant phagocytic leukocyte of the innate immune system, play an essential role in capturing and killing invading pathogens. Their antimicrobial repertoire includes the formation of Neutrophil Extracellular Traps (NETs), chromatin-based, web-like structures of DNA that facilitate the capture and killing of bacteria. In sepsis, however, it has been suggested that the uncontrolled release of NETs worsens disseminated coagulation and promotes venous thrombosis. Here, we describe how clinically relevant concentrations of the commonly used sedative propofol as well as a lipid composition similar to the propofol carrier impair NET production by human neutrophils. Drugs commonly administered in the Intensive Care Unit (ICU) may impact the inflammatory response to either worsen or improve clinical outcomes and may therefore be considered for additional therapeutic effects if clinical studies confirm such findings.

Chronic granulamatous disease: Two decades of experience from a paediatric immunology unit in a country with high rate of consangineous marriages.

Chronic granulomatous disease (CGD) is a primary immunodeficiency characterized by susceptibility to bacterial and fungal infections resulting from the inadequacy of phagocytic leucocytes to produce reactive oxygen radicals. CGD is a genetically heterogeneous disease with an X-linked recessive (XR-CGD) form caused by mutations in the CYBB (OMIM #300481) gene encoding the gp91(phox) protein, and an autosomal recessive (AR-CGD) form caused by mutations in the CYBA (OMIM #608508), NCF1 (OMIM #608512), NCF2 (OMIM #608515) and NCF4 (OMIM #601488) genes encoding p22(phox), p47(phox), p67(phox) and p40(phox), respectively. The genetic mutation of one of the cytosolic p47phox/p67phox proteins and membrane-bound gp91phox/p22phox proteins, which constitutes the NADPH oxidase enzyme complex, causes the disease. In this study, we evaluated the clinical, laboratory and genetic findings and the prognostic effects of molecular inheritance of our 24 CGD cases (14 XR, 10 autosomal recessive-AR). Consanguinity (three XR and all AR cases) showed statistically significant relationship with the type of hereditary inheritance (P<0.001). 83% patients had an infection since early infancy. The mean age of initiation of symptoms was earlier in XR cases, and 78% patients had respiratory tract infections. Bone marrow transplantation was performed in five XR cases (two ex) and four AR (one ex) cases. Three of nine XR and two of six AR cases deceased on medical follow-up. In countries especially with high consanguinity rates, the early diagnosis for appropriate prophylactic treatment of CGD is quietly important to avoid from recurrent severe infections, early death and fatal complications of late transplantation.

CYTOLOGICAL EXAMINATION OF EXPERIMENTAL PURULENT WOUNDS IN THE TREATMENT OF SILVER NANOPARTICLES IN ULTRASOUND CAVITATION

The treatment of purulent wounds is an important problem of modern surgery. Antibiotic resistance of bacteria dramatically reduces the effectiveness of traditional methods of treatment. Previous studies have indicated that silver nanoparticles have good antibacterial activity and do not cause bacterial resistance. Low-frequency ultrasound improves the bactericidal properties of nanoparticles, as well as plays an important role in cleaning wounds from purulent-necrotic tissue and delivering nanoparticles to the site of infection. The combination of the properties of silver nanoparticles and low-frequency ultrasound requires careful investigation in the treatment of purulent wounds. The purpose of the study was to substantiate the effectiveness of the treatment of purulent wounds with silver nanoparticles and low-frequency ultrasound by cytological examination. Materials and Methods. This study was carried out on 60 laboratory rats, which were equally divided into 3 groups. In the first group, treatment was carried out by low-frequency ultrasound; in the second group, a solution of silver nanoparticles was used together with low-frequency ultrasound; in the third, control group, a 0.05% Chlorhexidine solution was used. Silver nanoparticles with a size of 10-60 nm were synthesized by the polyol method. Results. The study showed that there were no statistically significant differences between the ultrasound, silver nanoparticles/ultrasound and Chlorhexidine groups on the first day. On the third day, a statistically significant increase in phagocytic neutrophilic leukocytes and fibroblasts was observed in the silver nanoparticles/ultrasound group compared with the first day. In comparison with the Chlorhexidine group, the number of monocytes (2.2 and 4.2 times) and macrophages (1.4 and 1.9 times) increased in the ultrasound and silver nanoparticles/ultrasound groups respectively, and the necrotic type of cytograms was not determined. On the seventh day, granulation tissue began to appear in the silver nanoparticles/ultrasound group, the percentage of leukocyte destruction decreased (by 4.5 times), microorganisms were almost not detected, regenerative types of cytograms appeared for the first time. The number of neutrophils in the control group exceeded the analogous parameter of the ultrasound group by 2.4 times and the silver nanoparticles/ultrasound group by 3.8 times. The number of fibroblasts became significantly larger in the ultrasound (2.2 times) and the silver nanoparticles/ultrasound (2.3 times) groups, compared with the group where Chlorhexidine was used. On the tenth day, the number of fibroblasts and cells of the monocytic-macrophage series increased in the control group, which indicates later regenerative processes. On the tenth day, there was a complete epithelization of wounds in the silver nanoparticles/ultrasound group, while healing occurred on day 12 in the ultrasound group, and on day 21 in the control group. Conclusions. The combined use of silver nanoparticles and low-frequency ultrasound significantly improves the cytological parameters of wound healing of purulent wounds and has clear advantages over the ultrasound monotherapy and the use of Chlorhexidine. The presented method reduces the treatment time and can be prospectively introduced into surgical practice.

CONDITION OF FAGOCYTOSIS OF EXPERIMENTAL ANIMALS WITH PERIODONTITIS DUE TO MODIFIED REACTIVITY

Introduction: Taking into account the patho-immune mechanisms of formation of inflammatory process in periodontium, there is a necessity for in-depth study of the pathogenesis of periodontal diseases from the position of changes in the reactivity of the organism. The aim: Carrying out a comparative estimation of phagocytic activity of leukocytes of animals with inflammation in periodontium with altered reactivity. Materials and methods: Experiments were performed on 30 white rats: Group I - 10 white rats with hypoergic reaction; Group ІІ - 10 white rats with hyperergic reaction; Group III - 10 white rats with normergic reaction - control group. Slaughter and blood sampling under thiopental anesthesia was performed 7 days after the beginning of the experiment. The percentage of phagocytic leukocytes - phagocytic index, phagocytic number were determined and the index of phagocytic activity was calculated. Results: The phagocytic index (Fi,%) decreased by 2,09 times (p <0,05) at hypoergic, it increased by 1,37 times (p <0,05) at hyperergic; the index of phagocytic activity (IFA) decreased by 1.96 times at hypoergic (p <0.05); growth was 1.94 times (p <0.05) in the hyperergic group of animals. In both experimental groups, the phagocyte number (Fu) increased by 6.25% and 41.7%, with hypoergic and hyperergic, respectively. Conclusions: Different directions of changes of the phagocytic activity were observed: increase of these parameters at hyperergic and decrease at hypoergic. An increase in phagocyte number was likely to indicate some autonomy of the process of phagocytosis and independence from the reactivity of the organism.

Human Beta Defensin 1 (hbd1) Levels in Sputum and Lysate of Mononuclear Blood Cells of Drug-Sensitive and Drug-Resistant Pulmonary Tuberculosis Patients Attending a Tertiary Hospital in Ibadan, Nigeria.

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