Activity Of Polymorphonuclear Leukocytes Research Articles

Effects of Butterfly Pea Extracts on Phagocytic Activity of Blood Polymorphonuclear Leukocytes and Muscular Lipid Peroxidation in Rabbits.

Sixteen 35-day-old male crossbred rabbits (New Zealand white × Thai native breed) with an initial weight of 484 ± 11.3 g were randomly divided into two groups of eight, constituting control and treatment groups. The treatment group was orally administered a crude extract of butterfly pea (Clitoria ternatea L.) at 0.5 g/kg body weight from weaning (at 35 days) to slaughter (at 90 days). The effects on the phagocytic activity of blood polymorphonuclear leukocytes, serum biochemistry, meat quality, muscular lipid peroxidation, the apparent digestibility of dry matter and nutrients, and gut histology were studied. The results revealed that the phagocytic function of circulating leukocytes (75 and 90 days) and alveolar macrophages (90 days) did not differ between the two groups. At slaughter, treated rabbits had lower blood urea nitrogen concentrations and higher liver weight than control rabbits (p < 0.05). After chilling at 4 °C for 24 h, a lower meat pH and the alteration of meat color (brighter, less yellow, lower hue angle, and decreased color saturation) were observed in the treated group (p < 0.05). Furthermore, lipid peroxidation (measured at 3, 5, and 7 storage days) in the meat of treated rabbits was lower than in controls (p < 0.05). The apparent digestibility of organic matter and ether extract (analyzed at 46 days for 4 days) was improved in the treated group (p < 0.05), whereas gut histology was unaffected. In conclusion, butterfly pea extract supplementation did not affect phagocytic function but led to a modification in meat color, delayed lipid peroxidation, and improved digestibility.

Клинико-лабораторные исследования в комплексной диагностике и лечении увеитов у детей

This work is devoted to the analysis of the effectiveness of new clinical and laboratory studies and their possible application in treatment of uveitis in children. The paper assesses the study of the glutathione-reducing system as a factor in the vulnerability of eye tissues to the effects of free radicals and peroxidation products during inflammation. The dependence of the clinical picture of uveitis and the frequency of detection of eye tissue antigens in the blood was analyzed to assess the localization and stage of the pathological process, which will undoubtedly determine the tactics of the therapy. Evaluation of the main indicators of the activity of peroxidation processes in uveitis is proposed to be carried out according to the original method for determining the functional activity of polymorphonuclear leukocytes (PNLs). It was proved that there is a close relationship between the level of PNLs activity, the level of intensity of peroxide processes and the ophthalmic picture of uveitis. Based on the results obtained in the following work, the use of pharmaceutical antioxidants, both traditional and those known drugs in which antioxidant properties were discovered for the first time, was proposed. This paper offers a concept of the development of toxic-allergic uveitis, including successive stages of the formation of immune complexes, inflammation, an increase in the functional activity of PMNs, their chemotaxis into the inflammation zone, infiltration through the capillary walls in the eye tissue, which leads to an avalanche-like increase in the level of oxidants, depletion of endogenous reserves of antioxidants, damage to cell membranes. Understanding these processes serves as an explication for developing optimal treatment regimens for uveitis and other inflammatory ophthalmpathology. Keywords: uveitis, retinal antigens, glutathione system, antioxidants, polymorphonuclear leukocytes, peroxidation, reactive oxygen species

INDICATORS OF PHAGOCYTOSIS IN WOMEN WITH ACNE DURING COMPREHENSIVE TREATMENT THAT INCLUDED PRP- AND IPL-THERAPY

Introduction. Acne is one of the most common dermatological diseases. It may have a chronic course, leaving permanent marks, and in last years has been tending to have more and more severe clinical course with widespread skin lesions. According to recent studies, the development of acne is due to the combined effect of endogenous and exogenous factors, among which endocrine diseases, disorders of metabolic processes, reduced systemic immunity and phagocytic ability of mononuclear phagocytes and granulocytes at various stages of phagocytosis of pyogenic cocci, which contributes to more severe clinical course, and frequent relapse of this diseases. It was also proved that the intestinal microbiota plays an important role in the formation of homeostasis and immune response.&#x0D; The Objective of the study is to determine the evolution of phagocytosis indices in patients with acne under different comprehensive treatments, using oral antibiotics, immunotherapy (PRPtherapy), probiotics, low-dose birth control pills and IPL- therapy.&#x0D; Materials and methods. We observed 93 women with acne aged from 18 to 25 years old. In 19 (20.43%) patient mild acne was diagnosed, in 41 (44.09%) – moderate acne, in 33 (35.48%) persons – severe acne, 54 (58.06%) persons suffered from acne up to 1 year, 39 (41.93%) – from 1 to 3 years. To assess the state of phagocytosis in patients with acne vulgaris, we determined phagocytic activity (PA) and phagocytic index (PI) of polymorphonuclear leukocytes, nitro blue tetrazolium recovery test (NBT test spontaneous) and NBT-test pyrogenal stimulated by recognized methods.&#x0D; Results. Analysis of the studied parameters of phagocytosis at the end of treatment showed a significant increase in patients of the core group who were administered a comprehensive treatment which included oral antibiotic, probiotic, and low-dose birth control pills, PRP-therapy and IPLtherapy, as compared with the patients of other groups under study.&#x0D; Conclusions. Using combined therapy for women with acne occurring against the backdrop of a sluggish process of phagocytosis and concomittant intestinal dysbiosis leads to normalization of the leading indices of phagocytosis (PI, PA, NBT tests both spontaneous and stimulated), and enhances their phagocytic activity both during capture and formation of bactericidal activity and in the final stages of phagocytosis justifying the feasibility of a combined use of antibiotics, probiotic, low-dose birth control pills PRP-therapy and IPL-therapy in the treatment of acne.

Bacteria and Viruses in Periodontics- A review

Periodontal diseases are infectious diseases, but the specific mechanism by which the tooth-supportive tissue is destroyed is not clearly understood. Periodontitis is a multifactorial, chronic disease followed by destruction of encompassing structures of teeth and when left untreated leads to loss of alveolar bone and exfoliation of the involved teeth. The main etiological factor for development of periodontitis is oral biofilm containing anaerobic microorganisms. Microbiological culture studies have identified more than 1200 bacterial species in the oral cavity. Although the role of bacterial plaque in general seems to be evident, on the contrary the role of virus has been largely unexplored. Viral infection impairs periodontal defenses, thereby permitting subgingival overgrowth of periodontopathic bacteria. The role of viruses is significant, as they may induce abnormalities in the adhesion, chemotaxis, phagocytosis, and bactericidal activities of polymorphonuclear leukocytes. When associated with one another, viruses and bacteria have stronger periodonto-pathogenic potential than individually. Therefore, it is significant to know all etiologic factors and such an insight would lead to the better treatment of the disease.

Changes in the organism of patients with chronic inflammatory diseases of the upper respiratory tract at different periods after administration of influenza vaccine. Message 1. Status of congenital immune parameters after a single anti-influenza vaccination

Introduction: Studies of the clinical status, production of antibodies to hemagglutinins of influenza A and B viruses and the content of immune complexes in patients with chronic inflammatory diseases (CID) of the upper respiratory tract (URT) before and within 36 weeks of vaccination after parenteral vaccination Vaxigrip leads to a short-term (~ 6 months) increase in anti-influenza hemagglutinin t itres. According to modern ideas about the mechanisms of antiviral protection, in addition to humoral, mainly due to antibodies, antiviral protection is largely due to the activation of innate immune factors, both systemic and local. Aim: to evaluate in the dynamics of observations the influence of a single parenteral influenza vaccination on the factors of innate immunity in patients with chronic inflammatory diseases of the upper respiratory tract. Material and methods: Among the factors of innate immunity a significant role is played by interferons, the activity of natural cytolytic cells (NK) of various histogenesis, phagocytic cells and non-specific proteins such as defensins, elafins. Therefore, quantitative and functional characteristics of NK-cells, phagocytic absorption activity of polymorphonuclear leukocytes (PMN), systemic content of γ-interferon (IFN-γ) were studied in patients at the systemic level. At the local level, the concentrations of defensin-β and α-interferon were studied in mucosa secretes (MS) according to modern ideas about the state of the mechanisms of innate antiviral immunity. From the autumn-winter period of 2019 to November 2020, 32 donors (control group) and 32 patients with CID of both sexes were examined, 11 of whom were diagnosed with chronic rhinosinusitis, 9 with chronic tonsillitis, and 12 with chronic pharyngitis. No vaccinations were given 1 year before the study. Samples of venous serum and oropharyngeal secretes of all clinically healthy patients, as well as persons with CID were obtained at the initial examination. After parenteral vaccination with Vaxigrip (SANOFI PASTEUR, S.A., France), blood and MS were tested twice more in vaccinated patients: 12 and 36 weeks after vaccination. The obtained samples of experimental material were stored at a temperature of -20 C◦ until the simultaneous determination of innate immune factors (interferons), in particular with the help of reagents from Cytokine, Hema Medica (RF), defensin-β (Hucalt-bio, the Netherlands). Measurements were performed on a microplate enzyme-linked immunosorbent assay "STAT FAX 2100 (USA). Statistical processing was performed using the non-parametric criterion "U" - Mann-Whitney or Fisher's exact method. Differences were considered statistically significant at p &lt;0.05. The results were given in the form of arithmetic mean (M), median (Me) and percentiles (P25-P75). Results: In determining the role of innate immunity factors in vaccination with influenza vaccine, we conducted clinical and immunological examination of patients with chronic inflammatory diseases of the upper respiratory tract, which included systemic determination of γ-interferon, quantitative and functional activity of natural cytotoxic cells, phagocytosis activity at the stage of attraction and capture. Levels of α-interferon and defensin-β in oropharyngeal secretion (MS) were studied. It was found that before influenza vaccination in patients with chronic inflammatory diseases of the upper respiratory tract found an increased content of γ-interferon and decreased phagocytic activity of polymorphonuclear leukocytes. Decreased α-interferon and defensin-β concentrations were detected in MS patients before vaccination. After vaccination, there were positive changes in the direction of increasing the concentration of nonspecific local immunity factor – defensing 1β and partly in the observation period of 3 months after vaccination- α-interferon. Сonclusion: Among the factors of innate immunity in patients with CІD at the systemic level, decreased cytolytic activity of NK, increased levels of γ-interferon and decreased phagocytic function of neutrophils. Decreased concentration of α-interferon and defensin-1β is determined in the saliva of patients with CID. Vaccination of patients with chronic inflammatory diseases of the upper respiratory tract with influenza vaccine has a positive effect on the state of innate immune factors of local and systemic level: concentration of γ-interferon, phagocytic activity of neutrophils and enhancing the functional activity of natural killer cells, increase in saliva of defensin-1β.

Inhibition of Peroxiredoxin 6 PLA2 Activity Decreases Oxidative Stress and the Severity of Acute Lung Injury in the Mouse Cecal Ligation and Puncture Model

IntroductionALI is associated with bacterial infection (sepsis), either as an initiating factor or as a secondary complication. Activation of polymorphonuclear leucocytes (PMN) recruited into the lungs causes the release of free radicals and reactive oxygen species (ROS) that are responsible for many of the clinical manifestations of ALI. Since NADPH oxidase (NOX2) is the major source of ROS (i.e., superoxide anion) in PMN, NOX2 blockade has been proposed for the treatment of Acute Lung Injury (ALI). We reasoned that NOX2 blockers would be counter‐productive in ALI associated with bacterial infection by interfering with the bactericidal activity of PMN.HypothesisPhospholipase A2 inhibitory peptide (PIP‐2) that inhibits NOX2 activation (by preventing activation of Rac, a necessary NOX2 co‐factor) would inhibit the bactericidal activity of PMN and thereby limit clearance of bacteria from the lung; we posited that this effect of PIP‐2 could be overcome by the administration of appropriate antibiotics.MethodsUsing the cecal ligation and puncture (CLP) model of ALI associated with sepsis, we investigated the effect of inhibiting NOX2derived ROS production by administration of PIP‐2.ResultsThe number of colony‐forming bacteria in lungs and peritoneal fluid of mice with CLP approximately doubled at 18 h after intravenous treatment with 2 µg PIP‐2 /gram body weight (wt). Treatment with 10 µg PIP‐2/g body wt resulted in 100% mortality within 18 hr. Antibiotic treatment abolished both the increase in lung bacteria with low dose PIP‐2 and the increased mortality with high dose PIP‐2. Treatment with PIP‐2 in the presence of antibiotics resulted in significantly improved lung histology, decreased PMN infiltration, decreased lung fluid accumulation, and decreased oxidative lung injury compared to antibiotics alone.ConclusionAdministration of PIP‐2 provides partial protection against lung injury in a model of ALI due to bacterial infection while concurrent antibiotic treatment abolishes the deleterious effects of PIP‐2 on lung bacterial clearance. These results suggest that addition of PIP‐2 to the antibiotic regimen would be beneficial for treatment of ALI associated with bacterial infection.

Distinct behavior of bovine-associated staphylococci species in their ability to resist phagocytosis and trigger respiratory burst activity by blood and milk polymorphonuclear leukocytes in dairy cows

Mastitis affects a high proportion of dairy cows and is still one of the greatest challenges faced by the dairy industry. Staphylococcal bacteria remain the most important cause of mastitis worldwide. We investigated how distinct staphylococcal species evade some critical host defense mechanisms, which may dictate the establishment, severity, and persistence of infection and the outcome of possible therapeutic and prevention interventions. Thus, the present study investigated variations among distinct bovine-associated staphylococci in their capability to resist phagocytosis and to trigger respiratory burst activity of blood and milk polymorphonuclear neutrophil leukocytes (PMNL) in dairy cows. To do so, PMNL of 6 primiparous and 6 multiparous dairy cows were used. A collection of 38 non-aureus staphylococci (NAS) and 12 Staphylococcus aureus were included. The phagocytosis and intracellular reactive oxygen species (ROS) production by blood and milk PMNL were analyzed by flow cytometry. Phagocytosis, by both blood and milk PMNL, did not differ between S. aureus and NAS as a group, although within-NAS species differences were observed. Staphylococcus chromogenes (a so-called milk-adapted NAS species) better resisted phagocytosis by blood PMNL than the so-called environmental (i.e., Staphylococcus fleurettii) and opportunistic (i.e., Staphylococcus haemolyticus) NAS species. Otherwise, S. haemolyticus was better phagocytosed by blood PMNL than S. aureus, S. fleurettii, and S. chromogenes. No influence of the origin of the isolates within the staphylococci species in the resistance to phagocytosis by blood and milk PMNL was found. Overall, both S. aureus and NAS did not inhibit intracellular ROS production in blood and milk PMNL. Non-aureus staphylococci induced fewer ROS by milk PMNL than S. aureus, which was not true for blood PMNL, although species-specific differences in the intensity of ROS production were observed. Staphylococcus chromogenes induced more blood PMNL ROS than S. fleurettii and S. haemolyticus, and as much as S. aureus. Conversely, S. chromogenes induced fewer milk PMNL ROS than S. aureus. The origin of the isolates within the staphylococci species did not affect the ROS production by blood and milk PMNL. In conclusion, our study showed differences in staphylococci species in evading phagocytosis and triggering ROS production, which may explain the ability of some staphylococci species (i.e., S. aureus and S. chromogenes) to cause persistent infection and induce inflammation.

Inhibition of Peroxiredoxin 6 PLA2 Activity Decreases Oxidative Stress and the Severity of Acute Lung Injury in the Mouse Cecal Ligation and Puncture Model

The use of agents to inhibit the production of reactive oxygen species (ROS) has been proposed for the treatment of Acute Lung Injury (ALI). However, this approach also inhibits the bactericidal activity of polymorphonuclear leucocytes (PMN) and other cells, raising the possibility of aggravating lung injury in ALI associated with bacterial infection. We used the cecal ligation and puncture (CLP) model of ALI associated with sepsis to investigate the effect of inhibiting NADPH oxidase 2 (NOX2)-derived ROS production, the main source of ROS in lungs. A phospholipase A2 inhibitor called peroxiredoxin 6 inhibitory peptide-2 (PIP-2) was used to inhibit NOX2 activation; the peptide prevents liberation of Rac, a necessary NOX2 co-factor. At 18 h after intravenous treatment with 2 µg PIP-2 /gram body weight (wt), the number of colony-forming bacteria in lungs and peritoneal fluid of mice with CLP was approximately doubled as compared to untreated mice. Treatment with 10 µg PIP-2/g body wt resulted in 100% mortality within 18 h. Antibiotic treatment abolished both the increase in lung bacteria with low dose PIP-2 and the increased mortality with high dose PIP-2. Treatment with PIP-2 plus antibiotics resulted in significantly improved lung histology, decreased PMN infiltration, decreased lung fluid accumulation, and decreased oxidative lung injury compared to antibiotics alone. We conclude that the administration of PIP-2 provides partial protection against lung injury in a model of ALI due to bacterial infection, while concurrent antibiotic treatment abolishes the deleterious effects of PIP-2 on lung bacterial clearance. These results suggest that addition of PIP-2 to the antibiotic regimen is beneficial for treatment of ALI associated with bacterial infection.

COMPARATIVE EFFECTIVENESS OF VARIOUS SCHEMES OF IMMUNOREHABILITATION IN INFERTILITY OF TUBOPERITONEAL GENESIS

The study aimed to compare the effectiveness of various pharmacotherapy regimens for infertility of tubo-peritoneal genesis. Under constant supervision were 96 patients referred to the hospital for diagnostic laparoscopy for infertility of tubo-peritoneal genesis, divided equally into 4 groups depending on the pharmacological treatment methods: the 1st group received basic pharmacotherapy (BPT) after endoscopic surgery (antibacterial, antifungal, vitamin therapy). Patients of groups 2-4, in addition to BPT, received Hepon, Cycloferon or Lavomax, respectively. The control group consisted of 38 gynecologically healthy women. Laboratory examination was performed within 24 hours after the operation and on the 30th day after BPT. Vaginocervical lavage and plasma were assayed for the activity of lipid peroxidation processes, the state of the antioxidant system, the level of stable nitric oxide metabolites, neopterin, C-reactive protein, cytokines (TNFα, IL-1β, IL-8, IFNγ, IL-18, G-CSF, IL-4, IL-10), immunoglobulins (IgM, IgG, IgA, sIgA), components of the complement system (C3, C4, C5, C5А), phagocytic and oxygen-dependent activity of polymorphonuclear leukocytes. It was established that the use of immunomodulatory and antiviral activity medication with BPT according to the degree of increasing efficiency in the correction of immunometabolic laboratory parameters at the systemic and local level in infertility of tuboperitoneal genesis is as the following sequence: basic pharmacotherapy &lt; basic pharmacotherapy + Hepon &lt; basic pharmacotherapy + Cycloferon &lt; basic pharmacotherapy + Lavomax.

The possibility of polymorphonuclear leukocyte activation in dental socket healing by freeze-dried Aloe vera induction

Background: Research has proved that Aloe vera has anti-inflammatory, anti-bacterial, and immunomodulator properties that can accelerate the healing process. These properties could be beneficial in the tooth extraction wound-healing process. Neutrophils are the first inflammatory cells to invade and are the predominant cell markers in the wound for 24 hours after injury. Polymorphonuclear leukocytes (PMN) provide initial protection against micro-organisms, as they engulf and digest foreign bodies. Purpose: The aim of this study was to prove the effect of Aloe vera 90% gel application on the PMN count in the healing process of Cavia cobaya following tooth extraction. Methods: This laboratory experimental research was done using Post-Test Only Control Group Design. The lower incisive of twelve male Cavia cobayas were extracted then divided into four groups. Group P1 and P3 had 90% Aloe vera gel applied into the socket and sutured; group K1 and K3, received no 90% Aloe vera gel application and acted as controls. Samples from group P1 and K1 were terminated and had their mandibula taken one day after tooth extraction, and those from group P3 and K3 were terminated after three days. Tissues were then processed into histology slides, and PMN cells were counted. Results: There were significant differences in the PMN count between groups K1, P1, and K3 compared with P3 (p &lt; 0.05). Higher PMN count was shown on the group containing Aloe vera 90% gel compared with control group, both on day one and three after tooth extraction. Conclusion: Aloe vera 90% gel can increase the PMN count in the inflammation phase of the tooth extraction wound-healing process.

Engineered nanoceria modulate neutrophil oxidative response to low doses of UV-B radiation through the inhibition of reactive oxygen species production.

To avoid aging and ultraviolet mediated skin disease the cell repair machinery must work properly. Neutrophils, also known as polymorphonuclear leukocytes, are the first and most abundant cell types which infiltrate sites of irradiation and play an important role in restoring the microenvironment homeostasis. However, the infiltration of neutrophils in ultraviolet-B (UV-B) irradiated skin might also contribute to the pathophysiology of skin disease. The polymorphonuclear leukocytes activation induced by UV-B exposure may lead to prolonged, sustained NADPH oxidase activation followed by an increase in reactive oxygen species (ROS) production. Our previous work showed that cerium oxide nanoparticles can protect L929 fibroblasts from ultraviolet-B induced damage. Herein, we further our investigation of engineered cerium oxide nanoparticles (CNP) in conferring radiation protection specifically in modulation of neutrophils' oxidative response under low dose of UV-B radiation. Our data showed that even low doses of UV-B radiation activate neutrophils' oxidative response and that the antioxidant, ROS-sensitive redox activities of engineered CNPs are able to inhibit the effects of NADPH oxidase activation while conferring catalase and superoxide dismutase mimetic activity. Further, our investigations revealed similar levels of total ROS scavenging for both CNP formulations, despite substantial differences in cerium redox states and specific enzyme-mimetic reaction activity. We therefore determine that CNP activity in mitigating the effects of neutrophils' oxidative response, through the decrease of ROS and of cell damage such as chromatin condensation, suggests potential utility as a radio-protectant/therapeutic against UV-B damage.

Pathogenic Effect of Prevotella intermedia on a Mouse Pneumonia Model Due to Methicillin-Resistant Staphylococcus aureus With Up-Regulated α-Hemolysin Expression.

Background: Methicillin-resistant Staphylococcus aureus (MRSA) is a common causative agent of pneumonia; however, the detailed mechanism underlying severe MRSA pneumonia, including association with oral hygiene or periodontitis, remains poorly characterized. In this study, we examined the pathogenic effect of Prevotella intermedia, a major periodontopathic pathogen, on MRSA pneumonia.Methods: The pathogenic effect of the supernatant of P. intermedia (Pi Sup) was investigated in a murine MRSA pneumonia model, using several clinical strains; whereas the bactericidal activity of polymorphonuclear leukocytes (PMNs) was investigated in vitro. The effect of Pi Sup on messenger RNA (mRNA) expression of the toxin/quorum sensing system (rnaIII) was investigated by quantitative reverse transcription PCR both in vitro and in vivo.Results: Mice infected by hospital-acquired MRSA (HA-MRSA) with Pi Sup exhibited a significantly lower survival rate, higher bacterial loads in the lungs, and higher α-hemolysin (hla) expression in the lungs, than those without Pi Sup. A similar effect of Pi Sup was not observed with MRSA strains producing Panton-Valentine leucocidin (PVL) or toxic shock syndrome toxin (TSST). In vitro, Pi Sup suppressed bactericidal activity of PMNs against the HA-MRSA strain. HA-MRSA was the clinical strain with the highest ability to proliferate in the lungs and was accompanied by time-dependent up-regulation of rnaIII and hla.Conclusions: Our results provide novel evidence that the product of P. intermedia exerts a pathogenic effect on MRSA pneumonia, in particular with a strain exhibiting strong proliferation in the lower airway tract. Moreover, our results indicate that P. intermedia affects MRSA toxin expression via quorum sensing in a strain-dependent fashion, which might be important for understanding the pathogenesis of severe MRSA pneumonia.

Stimulation of polymorphonuclear leukocyte activity under the influence of low strength electric field

It is assumed that electrical stimulation can be the most important physical effect providing regulation of the activity of immune cells such as polymorphonuclear leukocytes (PMNL). To study the effect of electrical stimulation on PMNL activity, the experimental setup based on electric signal generator and biochemiluminometer was developed, which allows performing electrostimulation of cells and studying the generation of reactive oxygen species (ROS) in PMNL simultaneously. Impact on the cells was carried out using electrodes immersed in incubation medium connected to electric signal generator. Non-uniform electric field was established, the intensity of which decreased with distance and time that imitated the characteristics of endogenous electric fields in places of inflammation in organism. During stimulation, the field strength was low, up to 1 V/cm. It was found that as a result of electrical stimulation, a change in the functional capabilities of human blood PMNL is observed. The severity of effects depends on the voltage applied to the electrodes and the duration of the observation. The key functional processes in PMNL such as ROS generation and myeloperoxidase enzyme secretion are enhanced in the initial period of electric field influence. A few hours after electrical stimulation, inhibition of PMNL activity and the gradual destruction of cells are observed. The obtained data testify the regulatory role of electrical stimulation in relation to PMNL activity which necessary for the implementation of the bodyʼs immune defense.

Labdane diterpenoids from Forsythia suspensa with anti-inflammatory and anti-viral activities

Five previously undescribed labdane diterpenoids, named Forsypensins A-E, were isolated from the fruits of Forsythia suspensa. The structures and relative configurations of the compounds were elucidated via extensive spectroscopic methods, and their absolute configurations were fully confirmed by single crystal X-ray diffraction analyses using Cu Kα radiation and electronic circular dichroism data. The five labdane diterpenoids showed in vitro anti-inflammatory activity in rat polymorphonuclear leukocytes, inhibiting the rates of β-glucuronidase release by 43.6%–49.2% at concentrations of 10 μM. The compounds also had anti-viral activity against influenza A (H1N1) virus and respiratory syncytial virus (RSV), with IC50 values in the range 21.8–27.4 μM, and EC50 values in the range 10.5–15.4 μM, respectively.

Neutrophils Enhance Their Own Influx to Sites of Bacterial Infection via Endosomal TLR-Dependent Cxcl2 Production.

The influx of neutrophils to infection sites is a fundamental step in host defenses against the frequent human pathogen group B Streptococcus (GBS) and other extracellular bacteria. Using a mouse model of GBS-induced peritonitis, we show in this study that the chemokines Cxcl1 and Cxcl2 play distinctive roles in enhancing the recruitment and the antibacterial activities of neutrophils in a manner that is linked to differences in the cellular sources of these mediators. Cell depletion experiments demonstrated that neutrophils make a significant contribution to the in vivo production of Cxcl2 but not Cxcl1. In vitro, neutrophils responded weakly to LPS but released high levels of Cxcl2 after stimulation with GBS or other bacteria. Neutrophil-derived Cxcl2 acted in an autocrinous manner to increase its own production and to enhance antibacterial activities, including the release of oxygen radicals. In both neutrophils and macrophages, the production of Cxcl1/2 largely required the presence of functional UNC93B1, a chaperone protein involved in signaling by endosomal TLRs. Moreover, the phenotype of UNC93B1-defective phagocytes could be recapitulated by the simultaneous absence of TLR7, 9, and 13 but not by the absence of individual TLRs. Collectively, our data show that neutrophils recognize Gram-positive and Gram-negative bacteria by means of multiple phagosomal TLRs, resulting in de novo synthesis of Cxcl2, amplification of neutrophil recruitment, and potentiation of their antibacterial activities. These data may be useful to devise alternative therapeutic strategies aimed at enhancing the recruitment and the functional activities of polymorphonuclear leukocytes during infections caused by antibiotic-resistant bacteria.

Riskometric assessment of factors affecting population health in situational analysis features of cytochemical indicators of activity circulating and tissue leukocytes and oxidative stress as a factor of chronic inflammation

The study of the mechanism of oxidative stress and regulation of oxygen-dependent processes is important, as the establishment of a complex interaction between oxidative stress, oxidative protein damage and the body's antioxidant system makes it possible to clarify metabolic pathways of disease pathogenesis. In our study, we concluded that in patients with chronic non-specificinflammatory diseases of the genitals of men and women, the phagocytic activity of polymorphonuclear leukocytes is reduced mainly by the oxygen-dependent mechanism, and tissue - by the oxygen-independent. As an exception in patients with chlamydial infection, the phagocytic activity of tissue polymorphonuclear leukocytes was also reduced by an oxygen-dependent mechanism.

Inflammation and oxidative stress transcription profiles due to in vitro supply of methionine with or without choline in unstimulated blood polymorphonuclear leukocytes from lactating Holstein cows

Neutrophils are the most important polymorphonuclear leukocytes (PMNL), representing the front-line defense involved in pathogen clearance upon invasion. As such, they play a pivotal role in immune and inflammatory responses. Isolated PMNL from 5 mid-lactating Holstein dairy cows were used to evaluate the in vitro effect of methionine (Met) and choline (Chol) supplementation on mRNA expression of genes related to the Met cycle and innate immunity. The target genes are associated with the Met cycle, cell signaling, inflammation, antimicrobial and killing mechanisms, and pathogen recognition. Treatments were allocated in a 3 × 3 factorial arrangement, including 3 Lys-to-Met ratios (L:M, 3.6:1, 2.9:1, or 2.4:1) and 3 levels of supplemental Chol (0, 400, or 800 μg/mL). Three replicates per treatment group were incubated for 2 h at 37°C and 5% atmospheric CO2. Both betaine-homocysteine S-methyltransferase and choline dehydrogenase were undetectable, indicating that PMNL (at least in vitro) cannot generate Met from Chol through the betaine pathway. The PMNL incubated without Chol experienced a specific state of inflammatory mediation [greater interleukin-1β (IL1B), myeloperoxidase (MPO), IL10, and IL6] and oxidative stress [greater cysteine sulfinic acid decarboxylase (CSAD), cystathionine gamma-lyase (CTH), glutathione reductase (GSR), and glutathione synthase (GSS)]. However, data from the interaction L:M × Chol indicated that this negative state could be overcome by supplementing additional Met. This was reflected in the upregulation of methionine synthase (MTR) and toll-like receptor 2 (TLR2); that is, pathogen detection ability. At the lowest level of supplemental Chol, Met downregulated GSS, GSR, IL1B, and IL6, suggesting it could reduce cellular inflammation and enhance antioxidant status. At 400 µg/mL Chol, supplemental Met upregulated PMNL recognition capacity [higher TLR4 and L-selectin (SELL)]. Overall, enhancing the supply of methyl donors to isolated unstimulated PMNL from mid-lactating dairy cows leads to a low level of PMNL activation and upregulates a cytoprotective mechanism against oxidative stress. Enhancing the supply of Met coupled with adequate Chol levels enhances the gene expression of PMNL pathogen-recognition mechanism. These data suggest that Chol supply to PMNL exposed to low levels of Met effectively downregulated the entire repertoire of innate inflammatory-responsive genes. Thus, Met availability in PMNL during an inflammatory challenge may be sufficient for mounting an appropriate biologic response.

Carbon monoxide-releasing molecule-3 (CORM-3) offers protection in an in vitro model of compartment syndrome.

Limb compartment syndrome (CS), a complication of trauma, results in muscle necrosis and cell death; ischemia and inflammation contribute to microvascular dysfunction and parenchymal injury. Carbon monoxide-releasing molecule-3 (CORM-3) has been shown to protect microvascular perfusion and reduce inflammation in animal models of CS. The purpose of the study was to test the effect of CORM-3 in human in vitro CS model, allowing exploration of the mechanism(s) of CO protection and potential development of pharmacologic treatment. Confluent human vascular endothelial cells (HUVECs) were stimulated for 6h with serum isolated from patients with CS. Intracellular oxidative stress (production of reactive oxygen species (ROS)) apoptosis, transendothelial resistance (TEER), polymorphonuclear leukocyte (PMN) activation and transmigration across the monolayer in response to the CS stimulus were assessed. All experiments were performed in the presence of CORM-3 (100μM) or its inactive form, iCORM-3. CS serum induced a significant increase in ROS, apoptosis and endothelial monolayer breakdown; it also increased PMN superoxide production, leukocyte rolling and adhesion/transmigration. CORM-3 completely prevented CS-induced ROS production, apoptosis, PMN adhesion, rolling and transmigration, while improving monolayer integrity. CORM-3 offers potent anti-oxidant and anti-inflammatory effects, and may have a potential application to patients at risk of developing CS.

Influence of «Melanizol» and «Klimedeks» pessaries on phagocytic activity of polymorphonuclear leukocytes in vitro

Local immunity during aerobic vaginitis is provided by the functioning of the mucous membranes, the secretory immune system and cells that have the ability to phagocytosis. Neutrophils related to polymorphonuclear leukocytes (PNL), involved in phagocytosis reactions, are markers of inflammation and prevail in the vagina of patients with an infection.&#x0D; The aim of the work was to study the effect of new vaginal pessaries «Melanizol» and «Klimedeks» on the phagocytic activity of PNL in vitro.&#x0D; The study of the influence of «Melanizol» and «Klimedeks» pessaries on the activity of PNL to phagocytize cells of yeast in vitro has been investigated. Reference drugs were «Gravagin» pessaries and vaginal tablets «Mikozhynax» and «Neotrizol». Determination of phagocytic activity (FA) of neutrophils was investigated according to generally accepted parameters: phagocytic index (FI) and phagocytic number (FN). Statistical processing was performed using the «Statistica 6.0» package using the Mann-Whitney Criterion the Mann-Whitney Criterion.&#x0D; FI significantly increased under the influence of pessaries «Kimedeks» and tended to increase under the influence of pessaries «Melanizol» (1.24 and 1.18 times, respectively) in comparison with intact samples. A significant increase in the FN under the influence of means «Melanizol» and «Klimedeks» was observed at 33% and 30%, respectively, during the study. «Melazinol» and «Klimedeks» pessaries significantly increased the FA in comparison with «Neotrizol» and «Gravagin» reference drags in vitro. The reference drug «Mikozhynax» did not have an effect on PNL in an experiment on the study of the effect on phagocytic activity in vitro.&#x0D; According to the results of in vitro studies, the pessaries «Melanizol» and «Klimedeks» influenced on the phagocytic activity of polymorphonuclear leukocytes, increased the phagocytic index and phagocytic number in comparison with intact samples. The studied pessaries were superior to the reference drugs «Gravagin», «Neotrizol» and «Mikozhynax». Thus, the investigated pessaries «Melanisol» and «Klimedeks» can be studied as drugs for the treatment of vaginitis with a reduced immune status.

The dual functionality of antimicrobial peptides Os and Os-C in human leukocytes.

Antimicrobial peptides (AMPs), Os and Os-C, have been identified as multifunctional peptides with antibacterial, antiendotoxin, and anti-inflammatory properties. For further development of Os and Os-C as therapeutic peptides, it is essential to evaluate these effects in human mononuclear (MN) and polymorphonuclear (PMN) leukocytes. The cytotoxicity and the effects of both peptides on MN and PMN morphology were determined with the Alamar-Blue assay and scanning electron microscopy, respectively. The ability of Os and Os-C to induce reactive oxygen species (ROS) and to protect against 2,2'-azobis(2-amidinopropane) dihydrochloride-induced oxidative damage in both cell populations was evaluated using 2',7'-dichlorofluorescin diacetate (DCFH-DA). Using fluorescently labeled peptides, the ability of the peptides to cross the cell membranes of MN and PMN was also evaluated. At the minimum bactericidal concentrations of Os and Os-C, neither peptide was cytotoxic. Os caused morphological features of toxicity at 100μM, entered MN cells, and also protected these cells against oxidative damage. Os-C caused MN and PMN leukocyte activation associated with ROS formation and was unable to penetrate cell membranes, indicating extracellular membrane interactions. This study confirms that both Os and Os-C at less than 100μM are not cytotoxic. The MN-specific uptake of Os identifies it as a cell-specific cargo-carrier peptide, with additional anti-inflammatory properties. In contrast, the ability of Os-C to activate MN and PMN cells implies that this peptide should be further evaluated as an AMP, which, in addition to its ability to eradicate infection, can further enhance host immunity. These novel characteristics of Os and Os-C indicate that these AMPs as peptides can be further developed for specific applications.