Leukocyte Phagocytosis Research Articles

Enniatin B alters bovine polymorphonuclear leukocytes phagocytosis and extracellular reactive oxygen species production in vitro.

Enniatins (ENNs) affect human and animal health. Different ENN analogs have been identified, but Enniatin B (ENN B) is the most detected in foods and feeds. This study investigated the effect of ENN B on bovine polymorphonuclear leukocytes (PMNs) challenged with increasing ENN B concentrations (0.625, 1.25, 2.5, 5, and 10μM). Bovine PMNs were isolated from the peripheral blood of dairy cows to evaluate the cell viability, chemotactic function, ability to phagocyte Gram+ and Gram- microorganisms, and extracellular Reactive Oxygens Species (ROS), with or without phorbol 12-myristate-13-acetate (PMA) as a pro-inflammatory challenge. Results demonstrated that ENN B did not affect bovine PMN viability and chemotactic activity at all concentrations (p=0.952; p=0.218, respectively). E. coli and S. aureus phagocytosis ability were reduced by ENN B at the highest concentrations (5 and 10μM) compared to the negative control (p≤0.001; p=0.001, respectively). Extracellular ROS production was increased by ENN B challenge under physiological and pro-inflammatory conditions (p=0.014; p<0.001, respectively). In conclusion, ENN B did not exert cytotoxic effects on bovine PMNs, while reduced phagocytic ability and increased the production of extracellular ROS, highlighting its potential role as an immunomodulator of the bovine innate immune response in vitro. IMPLICATIONS: Emerging mycotoxin Enniatin B is a common grain contaminant worldwide that can exert cytotoxic and immunotoxic effects in animal cells. We hypothesized that Enniatin B could in vitro affect the bovine immune response. In our study, Enniatin B did not affect bovine polymorphonuclear cell viability and chemotaxis, while a reduction of phagocytosis and a modulation of extracellular reactive oxygen species were observed. The present study shows that Enniatin B in vitro exerts a potential role as an immunomodulator of the bovine innate immune response putting animals at an increased risk of infection diseases.

Immunomodulation through the Circadian Clock: Impacts on Inflammation and Immunity

The circadian clock, an intrinsic timekeeping system regulating physiological functions over a 24-hour cycle, profoundly influences immune responses and inflammation. Immune cells, including neutrophils, macrophages, and lymphocytes, exhibit circadian rhythms in their numbers, function, and migration. These rhythms are regulated by both central and peripheral clocks, synchronizing immune cell activity with environmental cues. The circadian clock modulates key immune processes such as leukocyte trafficking, cytokine production, and phagocytosis, affecting the body’s ability to respond to pathogens and tissue injury. Furthermore, circadian rhythms tightly control inflammatory pathways, including NF-κB signaling, and regulate the balance between pro- and anti-inflammatory states in macrophages. Disruptions to circadian rhythms—due to factors like shift work or sleep disorders—are associated with heightened inflammation and increased risk for inflammatory diseases, such as autoimmune disorders, metabolic syndromes, and cancer. Understanding how circadian rhythms govern immune functions has significant implications for chronotherapy, which aims to optimize the timing of treatments to improve efficacy and reduce adverse effects. This review explores the mechanisms through which the circadian clock modulates immune responses and inflammation, highlighting the potential for circadian-based therapies in managing inflammatory and immune-related diseases. Keywords: Circadian clock, immunity, inflammation, immune cells, chronotherapy.

Colchicine can keep the viability of bacteria in mastitic milk by preventing leukocyte phagocytosis in dairy cow and goat.

Despite the occurrence of mastitis, no bacteria were detected in any of the milk samples after culture. This is partially because the neutrophils present in milk phagocytose bacteria during milk preservation. In this study, we investigated whether colchicine inhibited the decrease in viable bacteria in milk by suppressing phagocytosis during preservation. The number of viable bacteria decreased when cow milk was preserved for 5 h. However, the addition of 0.1 and 1% colchicine significantly increased the number of viable bacteria (p < 0.05). The percentage of culture-negative cow's milk increased more than two-fold after 5 h compared to that at 0 h of preservation, however this percentage was significantly reduced by the addition of colchicine (p < 0.05). When goat milk with mastitis was incubated with bacteria (Escherichia coli, Klebsiella pneumoniae, and Staphylococcus aureus), the percentage of phagocytosed neutrophils decreased significantly with the addition of colchicine (p < 0.05). These results indicate that colchicine suppressed the decrease in the number of viable bacteria by preventing neutrophil phagocytosis during milk preservation. These findings may help in the identification of mastitis-causing bacteria and the selection of antibiotics for the treatment of mastitis.

Impaired phagocytosis and oxidative respiratory burst activity in sickle cell anemia leukocytes

ObjectivesThis case-control study investigated the mode of leukocyte function in sickle cell anemia (SCA) to delineate the underlying immunopathology for early diagnosis and mitigate the increased bacterial infection risk in this patient population. MethodIn total, 90 participants comprising 24 hemoglobin (Hb)-AA, 22 Hb-AS, 23 steady state Hb-SS and 21 vaso-occlusive crisis state Hb-SS subjects were recruited for this study. The subjects were further divided into the following six groups: Hb-AA and Hb-AS subjects as control groups, Hb-SS subjects at steady state, Hb-SS subjects in a vaso-occlusive crisis state, Hb-SS subjects undergoing medication (Meds), and Hb-SS subjects undergoing medication plus blood transfusion (Meds/BT) group, respectively. Hematological analysis, Hb electrophoresis, leukocyte ratios, and leukocyte functional assays were assessed with standard methods, and interleukin 8 (IL-8) and L-selectin levels were evaluated using enzyme-linked immunosorbent assays. ResultsTotal leukocyte and monocyte counts were increased in the Hb-SS groups compared to the control groups. However, the Hb-SS groups had lower lymphocyte counts than the other groups (p < 0.005). Leukocyte viability was increased in the SCA groups, while phagocytic activities and oxidative respiratory burst were both reduced in the SCA groups (p < 0.005). Increased IL-8 levels were observed in all SCA groups (p < 0.05), whereas L-selectin levels of the Hb-SS steady and Hb-SS on Meds groups were decreased compared to the other groups (p < 0.05). The neutrophil-to-lymphocyte ratio, monocyte-to-lymphocyte ratio, and platelet-to-lymphocyte ratio were higher in the SCA groups than the control groups (p < 0.05). ConclusionImpaired leukocyte phagocytic and oxidative respiratory burst activities constitute altered leukocyte function in SCA, which can increase their susceptibility to infections and the risk of mortality, especially during the crisis state. Novel therapeutic approaches can be tailored specifically to enhance these leukocyte functions and mitigate the increased infection risk in SCA.

Exploring the Clinical Impact of Magnetite Nanoparticles in a Multifaceted Approach to Treating Multiple Sclerosis

Multiple sclerosis (MS) is a serious neurological disorder due to its widespread prevalence, chronic nature, frequent progression to disability, and tendency to affect young people. The pathogenesis of MS is based on the immunopathogenesis hypothesis. Biocompatible magnetite nanoparticles, which exhibit selective sorption activity towards cell membrane surface proteins, circulating immune complexes, lymphocytotoxic antibodies, and the complement system, as well as enhance phagocytic activity and the completion index of leukocyte phagocytosis, can be effectively used for immunocorrection. The main goal of the study is to slow the progression of MS, improve the neurological status and overall condition of the patient, and reduce the dynamics of the spread of demyelinating lesions in the brain. Materials and methods: A patient diagnosed with multiple sclerosis, secondary progressive type, cerebrospinal form, at the clinical aggravation stage was included in the study. Neurological status and disability were assessed using the EDSS scale, and brain MRI with contrast enhancement was performed. The nanodevice Micromage-B was administered orally as an immunosorbent and immunomodulator. The regimen and dosage of Micromage-B were personalized. Assessments of general condition and neurological status were conducted every 7 days for 6 months, with contrast-enhanced brain MRI performed in the 5th month. Results: The use of Micromage-B in MS treatment led to an objective improvement in neurological status, with reduced stiffness and fatigue in the lower extremities. Gait and coordination improved, hand tremors decreased, depression and concentration disorders subsided, appetite was restored, and speech improved. Throughout the treatment period, positive dynamics in the normalization of neurological status were observed. After 6 months, the total score on the EDSS scale decreased from 210 to 45. The most significant improvements were observed in the evaluation of the pyramidal system and coordination, with the EDSS Disability Scale score decreasing from 6.0 to 5.0. For the first time, contrast-enhanced brain MRI showed a reduction in the number of new demyelination foci by the 4th month of Micromage-B administration. The positive changes in neurological status correlated with the MRI results. The recovery of central nervous system activity in MS is likely not only due to the immunosuppressive properties of magnetite nanoparticles but also due to the activation of remyelination mechanisms and oligodendrocyte differentiation through enzymatic methylation. Conclusion: The use of biocompatible nanodevices in the complex treatment of MS is promising. Further improvement and study of the regimen and method of using biocompatible magnetite nanoparticles to enhance MS treatment effectiveness are required.

Effective Application of Biocompatible Magnetite Nanoparticles in the Treatment of Multiple Sclerosis: Results of a Clinical Study

Multiple sclerosis (MS) is a serious neurological disorder due to its widespread prevalence, chronic nature, frequent progression to disability, and tendency to affect young people. The pathogenesis of MS is based on the immunopathogenesis hypothesis. Biocompatible magnetite nanoparticles, which exhibit selective sorption activity towards cell membrane surface proteins, circulating immune com-plexes, lymphocytotoxic antibodies, and the complement system, as well as enhance phagocytic ac-tivity and the completion index of leukocyte phagocytosis, can be effectively used for immunocor-rection. The main goal of the study is to slow the progression of MS, improve the neurological status and overall condition of the patient, and reduce the dynamics of the spread of demyelinating lesions in the brain. Materials and methods: A patient diagnosed with multiple sclerosis, secondary progres-sive type, cerebrospinal form, at the clinical aggravation stage was included in the study. Neurologi-cal status and disability were assessed using the EDSS scale, and brain MRI with contrast enhance-ment was performed. The nanodevice Micromage-B was administered orally as an immunosorbent and immunomodulator. The regimen and dosage of Micromage-B were personalized. Assessments of general condition and neurological status were conducted every 7 days for 6 months, with contrast-enhanced brain MRI performed in the 5th month. Results: The use of Micromage-B in MS treatment led to an objective improvement in neurological status, with reduced stiffness and fatigue in the lower extremities. Gait and coordination improved, hand tremors decreased, depression and concen-tration disorders subsided, appetite was restored, and speech improved. Throughout the treatment period, positive dynamics in the normalization of neurological status were observed. After 6 months, the total score on the EDSS scale decreased from 210 to 45. The most significant improvements were observed in the evaluation of the pyramidal system and coordination, with the EDSS Disability Scale score decreasing from 6.0 to 5.0. For the first time, contrast-enhanced brain MRI showed a reduction in the number of new demyelination foci by the 4th month of Micromage-B administra-tion. The positive changes in neurological status correlated with the MRI results. The recovery of central nervous system activity in MS is likely not only due to the immunosuppressive properties of magnetite nanoparticles but also due to the activation of remyelination mechanisms and oligoden-drocyte differentiation through enzymatic methylation. Conclusion: The use of biocompatible nanodevices in the complex treatment of MS is promising. Further improvement and study of the regimen and method of using biocompatible magnetite nanoparticles to enhance MS treatment effec-tiveness are required.

An ex vivo Approach in European Seabass Leucocytes Supports the in vitro Regulation by Postbiotics of Aip56 Gene Expression of Photobacterium damselae subsp. piscicida.

Shewanella putrefaciens Pdp11 (SpPdp11) is a probiotic strain assayed in aquaculture; however, its postbiotic potential is unknown. Postbiotics are bacterial metabolites, including extracellular products (ECPs) that improve host physiology and immunity. Their production and composition can be affected by different factors such as the growing conditions of the probiotics. Photobacterium damselae subsp. piscicida strain Lg 41/01 (Phdp) is one of the most important pathogens in marine aquaculture. The major virulent factor of this bacterium is the exotoxin aip56, responsible for inducing apoptosis of fish leucocytes. Viable SpPdp11 cells have been reported to increase resistance to challenges with Phdp. This work aimed to evaluate the effect of two ECPs, T2348-ECP and FM1548-ECP, obtained from SpPdp11 grown under different culture conditions that previously demonstrated to exert different degradative and non-cytotoxic activities, as well as the effect on pathogens biofilm formation. These SpPdp11-ECPs were then analyzed by their effect on the viability, phagocytosis, respiratory burst and apoptogenic activity against European sea bass leucocytes infected or not with Phdp supernatant. Both ECPs, T2348-ECP and FM1548-ECP, were not cytotoxic against leucocytes and significantly reduced their apoptosis. Phagocytosis and respiratory burst of leucocytes were significantly reduced by incubation with Phdp supernatant, and not influenced by incubation with T2348-ECP or FM1548-ECP. However, both activities were significantly increased after leucocyte incubation with combined T2348-ECP and FM1548-ECP with Phdp supernatant, compared to those incubated only with Phdp supernatant. Finally, both T2348-ECP and FM1548-ECP significantly reduced the relative in vitro expression of the Phdp aip56 encoding gene.

Cyrtocarpa edulis fruit and its immunostimulant effect on Almaco Jack Seriola rivoliana: in vitro, in vivo and ex vivo studies.

The present study investigates for the first time chemical, proximate analyses and immunostimulant effect of Cyrtocarpa edulis fruit (CeF). Three design experiments were carried out to evaluate immunostimulant effect of C. edulis fruit: in vitro, in vivo and ex vivo studies in juveniles Almaco jack Seriola rivoliana. In general, nutraceutical studies performed by gas chromatography/mass spectrometry (GC-MS) in CeF revealed a major quantity of the carbohydrate groups and phytosterols such as β-sitosterol. Their phytochemical and antioxidant values exposed a significant content of total phenols, flavonoids, and tannins, showing an antioxidant capacity against hydroxyl and superoxide radical. The in vitro results confirm that CeF is edible and enhanced the innate immune response in head-kidney leukocytes after 24h of immunostimulation. The in vivo results showed that myeloperoxidase, nitric oxide production, as well as antioxidant enzymes were enhanced in skin mucus of those fish fed with CeF. Interestingly in the intestine, IL-β, TNF-α, MARCO and Piscidin gene expression were up-regulated in fish fed with C. edulis after 4weeks. Finally, ex vivo experiments showed an important enhancement on cellular parameters (phagocytosis, respiratory burst, myeloperoxidase, and nitric oxide production) in head-kidney leukocytes of fish fed CeF and intraperitoneally infected with A. hydrophila. The results demonstrate that C. edulis fruit (0.5%) represents an available phytochemical and antioxidant rich alternative with great potential as fish immunostimulant additive.

Hyperthermia-induced changes in leukocyte survival and phagocytosis: a comparative study in bovine and buffalo leukocytes.

Heat stress negatively affects health, welfare, and livestock productivity by impairing immune function, increasing disease incidence. In recent years, there has been increasing interest in understanding the immune system of water buffalo due to the growing economic impact of this species for the high quality and nutritional value of buffalo milk. While there are common responses across bovine and buffalo species, there are also some species-specific variations in the physiological responses to heat stress, mainly attributed to differences in metabolism and heat dissipation efficiency. At cellular level, the exposure to thermal stress induces several anomalies in cell functions. However, there is limited knowledge about the differential response of bovine and buffalo leucocytes to early and late exposure to different degrees of thermal exposure. The aim of this study was to compare the in vitro effect of hyperthermia on apoptosis and phagocytosis in leukocytes from bovine and buffalo species. For this, whole blood samples of six bovines and nine buffaloes were incubated at 39°C (mimicking normothermia condition) or 41°C (mimicking heat stress condition) for 1, 2, and 4 h. Two flow cytometric assays were then performed to evaluate apoptosis and determine functional capacity of phagocytic cells (neutrophils and monocytes). The results showed that the viability of bovine and buffalo leukocytes was differently affected by temperature and time of in vitro exposure. A higher percentage of apoptotic leukocytes was observed in bovines than in buffaloes at 39°C (3.19 vs. 1.51, p < 0.05) and 41°C (4.01 vs. 1.69, p < 0.05) and for all incubation time points (p < 0.05). In contrast, no difference was observed in the fraction of necrotic leukocytes between the two species. In both species, lymphocytes showed the highest sensitivity to hyperthermia, showing an increased apoptosis rates along with increased incubation time. In bovine, apoptotic lymphocytes increased from 5.79 to 12.7% at 39°C (p < 0.05), in buffalo, this population increased from 1.50 to 3.57% at 39°C and from 2.90 to 4.99% at 41°C (p < 0.05). Although no significant differences were found between the two species regarding the percentage of phagocytic neutrophils, lower phagocytosis capacity values (MFI, mean fluorescence intensity) were found in bovines compared with buffaloes at 41°C (27960.72 vs. 53676.45, p > 0.05). However, for monocytes, the differences between species were significant for both phagocytosis activity and capacity with lower percentages of bovine phagocytic monocytes after 2 h at 39°C and after 1 h at 41°C. The bovine monocytes showed lower MFI values for all temperature and time variations than buffaloes (37538.91 vs. 90445.47 at 39°C and 33752.91 vs. 70278.79 at 41°C, p < 0.05). In conclusion, the current study represents the first report on the comparative analysis of the effect of in vitro heat stress on bovine and buffalo leukocyte populations, highlighting that the leukocytes of buffalo exhibit relatively higher thermal adaptation than bovine cells.

The use of a Nanotechnological Drug (micromage-b) in the Complex Treatment of Multiple Sclerosis

Multiple sclerosis (MS) is a serious neurological problem because of its high prevalence, chronic course, frequent disability, and propensity to affect young people. The immunopathogenesis hypothesis underlies the origin of MS. Selective sorption activity of biocompatible magnetite nanoparticles against surface proteins of cell membranes, circulating immune complexes, lymphocytotoxin antibodies, complement system, the effect of increasing phagocytic activity and leukocyte phagocytosis completion index allows the effective use of these nanodevices for immunocorrection. The main goal of the study is to slow down the progression of MS, improve the neurological status and general condition of the patient, and reduce the dynamics of the spread of demyelinating foci in the brain.

Изучение состава фенольных соединений, антирадикальной активности и влияния на фагоцитоз сухого водного экстракта шишек сосны обыкновенной

Pine cone is a new promising type of plant raw material for using in a medical practice. Objective: determination of the phenolic compounds composition of pine cones dry aqueous extract, research of its toxicity, antiradical activity and effect on phagocytosis. Materials and methods. The research object was a collection of pine cones gathered in October 2020. A dry aqueous extract was obtained from the cones by extraction with hot water. The phenolic compounds composition of the pine cones dry aqueous extract was determined by ultra-efficient liquid chromatography/MS on a Waters Acquisition chromatograph with a diode-matrix UV detector and a tandem quadrupole MC detector TQD (Waters). To determine the antiradical activity, a reaction with a stable free radical, 2,2-diphenyl-1-picrylhydrazyl (DPPH), was used. The acute toxicity of the pine cones dry extract was researched by the V.V. Prozorovsky express method on white non-linear mice. The phagocytic activity of peripheral blood leukocytes was determined in vivo by a modified method. Results. The phenolic compounds composition of the extract was analysed. The research identified 12 substances present in the extract, representing at least 1% of the total area on the chromatogram. The following procyanidins were found in the pine cones extract: procyanidin B2, procyanidin B3, procyanidin C1, procyanidin D1. Additionally, flavonoids and phenolic carboxylic acids were identified. The pine cones dry extract demonstrated pronounced antiradical activity, comparable to that of ascorbic acid. The acute toxicity determination revealed that the LD50 of the researched compounds is in doses greater than 1260 mg/kg. Upon examination of the effect of pine cone extract on the phagocytic activity of peripheral blood leukocytes, an increase in the phagocytic index was observed, yet no change was noted in the activity of total leukocyte phagocytosis. Conclusion. The dry aqueous extract of pine cones contains biologically active components of phenolic nature, which suggests that further research into its pharmacological activity may be promising.

Anti-microbial activity and immunomodulation of recombinant hepcidin 2 and NK-lysin from flounder (Paralichthys olivaceus)

Infections due to pathogens impact global aquaculture economy, where diseases caused by bacteria should be in particular focus due to antibiotic resistance. Hepcidin and NK-lysin are important innate immune factors having potential to be exploited as alternatives to antibiotics due to their antimicrobial activity and immunomodulatory capacity. In this study, the recombinant proteins of hepcidin 2 and NK-lysin (rPoHep2 and rPoNKL) from flounder (Paralichthys olivaceus) were obtained via a prokaryotic expression system. The results exhibited that rPoHep2 and rPoNKL killed both gram-negative and gram-positive bacteria mainly via attachment and disruption of the membrane. Interestingly, both peptides could bind to bacterial DNA. The antiviral assay showed that both peptides have antiviral activity against hirame nonvirhabdovirus. They exhibited no cytotoxicity to the mammalian and fish cell lines. PoHep2 was found localized in G-CSFR-positive peritoneal cells. Moreover, rPoHep2 significantly enhanced the phagocytosis of flounder leukocytes in vitro. These findings suggested that neutrophils contained rPoHep2 and may respond to the immunoreaction of neutrophils. In summary, both rPoHep2 and rPoNKL possess antimicrobial activities and may be exploited to replace traditional antibiotics. rPoHep2 possess immune regulatory functions, that can be further investigated as an immunostimulant in aquaculture.

Modulatory Effect of Beneficial Enterococci and Their Enterocins on the Blood Phagocytes in Murine Experimental Trichinellosis.

Bacteriocins (enterocins) represent a new therapeutic strategy in various intestinal and non-intestinal infections. In antiparasitic defence, an oxidative inflammation of phagocytes is effective in destroying new-born Trichinella spiralis larvae. The strains Enterococcus faecium CCM8558 and E. durans ED26E/7 and their enterocins, enterocin M and a durancin-like enterocin, respectively, were administered daily, and mice were then infected with T. spiralis larvae on the seventh day of treatment. Phagotest and Bursttest kits were used to detect the phagocytosis and respiratory burst in blood leukocytes. T. spiralis infection inhibited phagocytosis from day 11 post-infection (dpi) during the migration of new-born larvae into the muscles. E. faecium CCM8558, E. durans ED26E/7, and the durancin-like enterocin increased phagocytic activity from day 11 dpi. Both strains and their enterocins (enterocin M and durancin-like) stimulated the ingestion capability of phagocytes from 18 to 32 dpi. Enterococci/enterocins therapy prevented a reduction in cells with respiratory burst caused by T. spiralis infection from 11 dpi. The enzymatic activity of phagocytes was stimulated on 18 and 25 dpi, particularly by E. faecium CCM8558 and enterocin M. Enterocin M and the durancin-like enterocin were as effective in stimulating phagocytosis as the bacterial strains that produce them. The stimulation of phagocytosis could contribute to decreased larval migration and reduced parasite burden in the host.

Clinical adaptation and the state of innate and humoral im- munity of premature infants with extremely low body weight who received oropharyngeal administration of colostrum

Breast milk is the optimal food for premature infants. However, the serious condition of premature newborns requires a complex of primary resuscitation care in the maternity unit, which makes it impossible to carry out early attachment to the breast and start breastfeeding. Currently, the neonatal community is actively discussing the immune effects of an alternative method of oropharyngeal administration of colostrum on the child’s adaptation to extrauterine existence. Purpose. To assess the clinical outcomes, the state of the monocytic immunity, the production of sIgA in coprofiltrates in premature infants with extremely low body weight who received colostrum in the first hours of life. Material and methods. 39 premature infants with extremely low body weight who received oropharyngeal administration of colostrum were examined. The expression of CD14+CD282+, CD14+CD284+, CD14+HLA-DR, CD14+CD64+, CD14+CD11b+, CD14+CD11c+ monocytes and the phagocytic ability of mononuclear cells in blood serum were determined by laser flow cytometry. The secretory IgA concentration was assessed in coprofiltrates. Results. In premature infants who did not receive oropharyngeal colostrum, there was an increase in leukocyte phagocytosis, the level of monocyte expression, which was due to an increased infectious morbidity. Mucosal immunity of children who received colostrum was characterized by an increased concentration of secretory IgA. Conclusion. It was found that children who received oropharyngeal administration of colostrum have a faster correction of transient hypoglycemia after birth, a shorter duration of parenteral nutrition, greater body weight at the time of discharge, and a decrease in the incidence of infectious pathology.

Antiphagocytic Properties of Polygallic Acid with Implications in Gouty Inflammation.

Polygallic acid (PGAL) has been used in vitro to protect synoviocytes from monosodium urate (MSU) crystals due to its anti-inflammatory properties. However, MSU crystals can also activate other cells of the synovial fluid (SF). We studied the impact of PGAL on the phagocytosis of MSU crystals, inflammation, and oxidative stress using an in vitro model with SF leukocytes and THP-1 monocyte cells. SF leukocytes were stimulated with PGAL and MSU crystals, proinflammatory cytokines and phagocytosis were assessed. In THP-1 cells, the effect of PGAL on the phagocytosis of MSU crystals and the levels of IL-1β, IL-6, TNF-α, and reactive oxygen species (ROS) was evaluated. PGAL was added to THP-1 cultures 24 h before MSU crystal addition as a pre-treatment, and IL-1β was measured. One-way ANOVA with Tukey's post hoc testwas performed, and a P value < 0.05 was considered statistically significant. PGAL (100 µg/mL) decreased phagocytosis in SF leukocytes by 14% compared to cells exposed to crystals without PGAL. In THP-1 cells, 100 and 200 µg/mL PGAL reduced phagocytosis by 17% and 15%, respectively. In SF cells, there was a tendency to decrease IL-1β and IL-6. In THP-1 cells, decreases in IL-1β and TNF-α, as well as a slight decrease in ROS, were identified. PGAL pre-treatment resulted in a reduction of IL-1β. PGAL inhibits MSU phagocytosis by exerting an anti-inflammatory effect on cells exposed to crystals. The use of PGAL before an acute attack of gout suggests an important protective factor to control the inflammation.

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.

Efficiency of Nanoparticles (Micromage-B) in the Complex Treatment of Multiple Sclerosis

Multiple sclerosis (MS) is a serious neurological problem because of its high prevalence, chronic course, frequent disability, and propensity to affect young people. The immunopathogenesis hypothesis underlies the origin of MS. Selective sorption activity of biocompatible magnetite nanoparticles against surface proteins of cell membranes, circulating immune complexes, lymphocytotoxic antibodies, complement system, the effect of increasing phagocytic activity and leukocyte phagocytosis completion index allows the effective use of these nanodevices for immunocorrection. The main goal of the study is to slow down the progression of MS, improve the neurological status and general condition of the patient, and reduce the dynamics of the spread of demyelinating foci in the brain. Materials and methods: a patient diagnosed with multiple sclerosis, secondary progressive type of course, cerebro-spinal form, clinical aggravation stage; EDSS neurological status and disability assessment scales; contrast-enhanced MRI of the brain. An oral form of the nanodevice Micromage-B was used as an immunosorbent and immunomodulator. The choice of the regimen and dosage of Micromage-B was personalized. Assessment of the general condition and neurological status was performed every 7 days for 6 months. Contrast-enhanced MRI of the brain was performed at the 5th month of the study. As a result of using Micromage-B in MS treatment, objective improvement of neurological status, reduction of stiffness and rapid fatigability of the lower extremities were observed. Gait and coordination improved, hand tremors decreased, depression and signs of concentration disorders disappeared, appetite restored, and speech improved. During the entire period of Micromage-B application, positive dynamics in the normalization of the neurological status was observed. After 6 months of treatment, the total score dropped by 165 to 35. It was found that the maximum positive effect was observed in the evaluation of the pyramidal system and coordination. The EDSS Disability Scale score decreased from 6.0 to 5.0. Contrast-enhanced MRI brain examination for the first time showed a decrease in the number of new foci of demyelination in the brain by the 4th month of Micromage-B administration. The positive dynamics of normalization of the neurological status correlated with the results of brain MRI. The process of recovery of central nervous system activity in MS is not only due to the immunosuppressive properties of magnetite nanoparticles, but is probably caused by the activation of remyelination mechanisms and oligodendrocyte differentiation through enzymatic methylation. Considering the above, the use of biocompatible nanodevices in the complex treatment of MS is a promising direction. The scheme and method of using biocompatible magnetite nanoparticles to improve the effectiveness of MS treatment require further improvement and study.

In vitro assessment of the genotoxicity and immunotoxicity of treated and untreated municipal effluents and receiving waters in freshwater organisms

Municipal wastewater effluent is one of the largest sources of pollution entering surface waters in the Laurentian Great Lakes. Exposure to wastewater effluent has been associated with impaired immune systems and induction of genotoxicity to aquatic animals. Due to habitat degradation and environmental pollution linked to industrial development and population growth, several regions of the Great Lakes have been designated Areas of Concern (AOCs). In this study, we assessed the effect of extracts of sewage influent, (treated) effluent and receiving surface waters from the Hamilton Harbour AOC and the Toronto and Region AOC (Ontario, Canada) on the phagocytic immune response of rainbow trout (Oncorhynchus mykiss) kidney leukocytes and the genotoxicity (DNA strand breaks) of these extracts on freshwater mussel (Eurynia dilatata) hemocytes. We identified and quantified numerous chemicals present in the various samples extracted for exposure. In freshwater mussels, extracts from Hamilton Harbour AOC induced DNA damage with the most frequency (12 out of 28 samples) regardless of sample type, reflecting past and present industrial activities. In contrast, extracts from Toronto and Region AOC induced DNA damage infrequently (2 out of 32 (summer) and 5 out of 32 (fall) samples, respectively) and from different WWTPs at different times. None of the extracts induced any significant effect on phagocytosis of rainbow trout kidney leukocytes. The present study indicates that despite overall improvements to effluent quality, treatment of influent by WWTPs may not result in a corresponding improvement of the genotoxicity of effluents. In vitro bioassays are useful and cost-effective rapid-screening tools for preliminary assessments of contamination of aquatic ecosystems.

Efficacy of a recombinant Lactobacillus plantarum Lp-pPG-Malt as an oral vaccine candidate against Aeromonas hydrophila infection in crucian carp

Aeromonas hydrophila (A. hydrophila), a gram-negative bacterium, causes serious diseases with various clinical symptoms in farm raised fish. Thus, different ways to prevent and control A. hydrophila infection need to be explored, including a vaccine. In this study, we evaluated the protective efficacy of an oral vaccine prepared from the A. hydrophila TPS maltoporin (Malt) with Lactobacillus plantarum (L. plantarum) against A. hydrophila infection in crucian carp (Carassius auratus). For the in vivo experiment, the oral vaccine was administered to crucian carp by feeding them fish diets containing Lp-pPG-Malt, Lp-pPG and PBS for 28 days. The enzyme-linked immunosorbent assay (ELISA), leukocyte phagocytosis assay and real-time quantitative polymerase chain reaction (RT-qPCR) were performed to measure the protective efficacy of the Lp-pPG-Malt. ELISA and leukocyte phagocytosis assay confirmed that Lp-pPG-Malt significantly enhanced the IgM level and nonspecific immune response of crucian carp compared with the control groups (Lp-pPG and PBS). The RT-qPCR results showed that the Lp-pPG-Malt increased the relative expression of immune-related genes (IL-10, IL-1β, TNF-α, IFN-γ) of crucian carp in various tissues (liver, spleen, head kidney and hind intestine). Moreover, Lp-pPG-Malt significantly increased the relative percent survival of fish after intraperitoneal injection with A. hydrophila (55%) compared with the Lp-pPG and PBS groups (0%). These findings suggest that Lp-pPG-Malt can serve as an oral vaccine candidate for A. hydrophila infection and that Malt can be used as an effective antigen in crucian carp farming.

Measurement of Calprotectin (S100A8/A9) in the Saliva of Pigs: Validation Data of A Commercially Available Automated Assay and Changes in Sepsis, Inflammation, and Stress.

Calprotectin (CALP, S100A8/A9), also named myeloid-related protein 8/14, is a dimer complex of S100A8 and S100A9 that belongs to the S-100 protein family. It is involved in inflammation and has a wide range of proinflammatory functions, such as cytokine production and regulation of leukocyte adhesion, migration, and phagocytosis. In humans, CALP traditionally can be measured in faeces, serum, and saliva as a biomarker of inflammation and sepsis. The objective of this study was to validate an automated assay for CALP measurements in the saliva of pigs, having the advantage of the use of a non-invasive sample that is easy to collect. The assay was precise and accurate. CALP in saliva measured by this assay showed significant changes depending on the hour of the day. It also showed significant increases in the saliva of pigs after the administration of lipopolysaccharide (LPS), and showed a rise, although with increases of lower magnitude, after a stressful stimulus. Further studies should be made to gain knowledge about the possible practical applications of the measurements of CALP in the saliva of pigs as a biomarker to evaluate the animals' health and welfare.