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In mammalian polymorphonuclear cells, polymorphonuclear structures are assembled by loop extrusion. Mega-loop anchors and interchromosomal hubs are stabilized by nuclear condensates. Diminished loop extrusion might alter nuclear deformability. Loop extrusion programs may instruct cellular architecture. It might be feasible to engineer artificial nuclear shapes and altered nuclear deformability to manipulate immune effector cell migration. Since the cardinal discovery by Metchnikoff that neutrophils assemble into polymorphonuclear structures, insights into the mechanisms that underpin the formation of such structures have remained rudimentary. Recent studies have demonstrated that in early mammalian hematopoiesis, mononuclear versus polymorphonuclear cell fate decisions are orchestrated by intricate competitive physical forces that involve loop extrusion and phase separation programs. Changing loop extrusion programs might enable the engineering of new nuclear shapes and artificial cytoplasmic architectures in effector immune cells and beyond. Immune cell fate decisions are regulated, at least in part, by nuclear architecture. Here, we outline how nuclear architecture instructs mammalian polymorphonuclear cell differentiation. We discuss how in neutrophils loop extrusion mechanisms regulate the expression of genes involved in phagocytosis and shape nuclear morphology. We propose that diminished loop extrusion programs also orchestrate eosinophil and basophil differentiation. We portray a new model in which competitive physical forces, loop extrusion, and phase separation, instruct mononuclear versus polymorphonuclear cell fate decisions. We posit that loop extrusion programs instruct the spatial organization of cytoplasmic organelles, including neutrophil granules, mitochondria, and endoplasmic reticulum. Finally, we suggest that changing loop extrusion programs might allow the engineering of new nuclear shapes and artificial cytoplasmic architectures. [ABSTRACT FROM AUTHOR]

The quality of used cooking oil can be improved by purifying it using bagasse activated charcoal. Purified used cooking oil can be used as raw material for soap. Use of soap can cause allergic contact dermatitis (ACD). Skin suffering from ACD is characterized by inflammatory cell infiltration of the skin, the inflammatory cells involved are mast cells, macrophages and polymorphonuclear cells (PMN), namely basophils, eosinophils and neutrophils. Therefore, inflammatory cells were chosen as a parameter for the ACD response to the use of used cooking oil liquid soap containing betel and aloe vera. Liquid soap formed was use an extract concentration of 4.5, 9 and 15 %. Skin sensitivity testing was tested on male Wistar rats aged 2 - 3 months for 14 days. Quantitative data was obtained through histopathological examination of the number of inflammatory cells, which were analyzed statistically. The results showed that the use of liquid soap with an extract concentration of 9 % (4.24 ± 0.74) and 15 % (3.96 ± 0.25) had no effect on basophil infiltration. The use of liquid soap with an extract concentration of 4.5 % (2.96 ± 1.17, 6.42 ± 0.86, 5.96 ± 0.38), 9 % (3.12 ± 0.47, 4.62 ± 0.68, 6.42 ± 1.06) and 15 % (3.36 ± 0.41, 5.08 ± 0.30, 6.44 ± 0.55) had no effect on infiltration of neutrophils, eosinophils and macrophages. The use of liquid soap with an extract concentration of 9 % (4.60 ± 0.86) had no effect on mast cell infiltration. Therefore, it can be concluded that the best concentration of 9 % extract betel and aloe vera showed no infiltration of inflammatory cells in rats' skin given soap from waste cooking oil. Thus, that the soap formed from waste cooking oil treated with bagasse activated charcoal and contain betel and aloe vera is predicted safe to used. [ABSTRACT FROM AUTHOR]

Neutrophils are dynamic cells, playing a critical role in pathogen clearance; however, neutrophil infiltration into the tissue can act as a double-edged sword. They are one of the primary sources of excessive inflammation during infection, which has been observed in many infectious diseases including pneumonia and active tuberculosis (TB). Neutrophil function is influenced by interactions with other immune cells within the inflammatory lung milieu; however, how these interactions affect neutrophil function is unclear. Our study examined the macrophage–neutrophil axis by assessing the effects of conditioned medium (MΦ-CM) from primary human monocyte-derived macrophages (hMDMs) stimulated with LPS or a whole bacterium (Mycobacterium tuberculosis) on neutrophil function. Stimulated hMDM-derived MΦ-CM boosts neutrophil activation, heightening oxidative and glycolytic metabolism, but diminishes migratory potential. These neutrophils exhibit increased ROS production, elevated NET formation, and heightened CXCL8, IL-13, and IL-6 compared to untreated or unstimulated hMDM-treated neutrophils. Collectively, these data show that MΦ-CM from stimulated hMDMs activates neutrophils, bolsters their energetic profile, increase effector and inflammatory functions, and sequester them at sites of infection by decreasing their migratory capacity. These data may aid in the design of novel immunotherapies for severe pneumonia, active tuberculosis and other diseases driven by pathological inflammation mediated by the macrophage–neutrophil axis. [ABSTRACT FROM AUTHOR]

ABSTRACT: Dry matter intake (DMI, kg/d) is closely related to the magnitude of negative energy and protein balance during the transition period, and the metabolic adaptations to support lactation in dairy cows. Thus, DMI might affect the development of cytological endometritis in the early postpartum period. Difficulty to adapt to these metabolic changes is related to impaired immune function and increased occurrence of reproductive disorders. We aimed to examine the association of pre- and postpartum DMI, body weight (BW), body condition score, milk yield and milk composition, and days to first ovulation with cytological endometritis at 15 (CYT15) and 30 DIM (CYT30). A second objective was to understand the association of vaginal discharge with CYT15 and CYT30 and performance. We conducted a pooled statistical analysis of 5 studies, including data from 280 multiparous Holstein cows. Based on the cutoffs for the percentage of uterine polymorphonuclear cells (PMN), determined by taking the median value of the data set for 15 and 30 DIM, cows were categorized as follows: LOW15 (PMN % at 15 DIM ≤24%; n = 125), HIGH15 (PMN % at 15 DIM >24%; n = 125), LOW30 (PMN % at 30DIM ≤7%; n = 141); and HIGH30 (PMN % at 30DIM >7%; n = 139). Cows in HIGH15 consumed an average of 1.97 ± 0.5 kg/d less DM than cows in LOW15 during prepartum, and 3.01 ± 0.5 kg/d less DM during postpartum. Dry matter intake (as a percentage of BW) was higher for cows in LOW15 during pre- and postpartum than for cows in HIGH15. Moreover, cows in HIGH15 tended to have lower milk yield than cows in LOW15 from the third until the fifth week postpartum. Although DMI was not associated with CYT30, DMI (as a percentage of BW) was lower for cows in LOW30 pre- and postpartum than for cows in HIGH30. There was no association between CYT30 and milk yield. Cows in LOW15 had greater days to first ovulation than cows in HIGH15, while cows in LOW30 also had greater days to first ovulation than cows in HIGH30. Simple regression analyses demonstrated linear associations of increased DMI, particularly postpartum, with decreased uterine PMN percentage and lower vaginal discharge score. Additionally, increased units of vaginal discharge score and increased percentage units of uterine PMN were linearly associated with decreased milk yield. Corroborating with the notion of the ovarian function being associated with uterine inflammatory status, cows in HIGH15 and HIGH30 ovulated on average 3 d before than cows in LOW15 and LOW30, respectively. Cytological endometritis at 15 DIM was associated with lower DMI from 4 wk before calving until 4 wk postpartum and was associated with lower milk yield. The association of vaginal discharge with cytological endometritis was variable and dependent on the day of evaluation.

Dry matter intake (DMI, kg/d) is closely related to the magnitude of negative energy and protein balance during the transition period, and the metabolic adaptations to support lactation in dairy cows. Thus, DMI might affect the development of cytological endometritis in the early postpartum period. Difficulty to adapt to these metabolic changes is related to impaired immune function and increased occurrence of reproductive disorders. We aimed to examine the association of pre- and postpartum DMI, body weight (BW), body condition score, milk yield and milk composition, and days to first ovulation with cytological endometritis at 15 (CYT15) and 30 DIM (CYT30). A second objective was to understand the association of vaginal discharge with CYT15 and CYT30 and performance. We conducted a pooled statistical analysis of 5 studies, including data from 280 multiparous Holstein cows. Based on the cutoffs for the percentage of uterine polymorphonuclear cells (PMN), determined by taking the median value of the data set for 15 and 30 DIM, cows were categorized as follows: LOW15 (PMN % at 15 DIM ≤24%; n = 125), HIGH15 (PMN % at 15 DIM >24%; n = 125), LOW30 (PMN % at 30DIM ≤7%; n = 141); and HIGH30 (PMN % at 30DIM >7%; n = 139). Cows in HIGH15 consumed an average of 1.97 ± 0.5 kg/d less DM than cows in LOW15 during prepartum, and 3.01 ± 0.5 kg/d less DM during postpartum. Dry matter intake (as a percentage of BW) was higher for cows in LOW15 during pre- and postpartum than for cows in HIGH15. Moreover, cows in HIGH15 tended to have lower milk yield than cows in LOW15 from the third until the fifth week postpartum. Although DMI was not associated with CYT30, DMI (as a percentage of BW) was lower for cows in LOW30 pre- and postpartum than for cows in HIGH30. There was no association between CYT30 and milk yield. Cows in LOW15 had greater days to first ovulation than cows in HIGH15, while cows in LOW30 also had greater days to first ovulation than cows in HIGH30. Simple regression analyses demonstrated linear associations of increased DMI, particularly postpartum, with decreased uterine PMN percentage and lower vaginal discharge score. Additionally, increased units of vaginal discharge score and increased percentage units of uterine PMN were linearly associated with decreased milk yield. Corroborating with the notion of the ovarian function being associated with uterine inflammatory status, cows in HIGH15 and HIGH30 ovulated on average 3 d before than cows in LOW15 and LOW30, respectively. Cytological endometritis at 15 DIM was associated with lower DMI from 4 wk before calving until 4 wk postpartum and was associated with lower milk yield. The association of vaginal discharge with cytological endometritis was variable and dependent on the day of evaluation. [Display omitted] [ABSTRACT FROM AUTHOR]

Aging is characterized by a loss of bone marrow hematopoietic tissue, systemic chronic inflammation, and higher susceptibility to infectious and noninfectious diseases. We previously reported the tightly regulated kinetics and massive daily production of neutrophils during homeostasis in adult rhesus macaques aged 3 to 19 yr (equivalent to approximately 10 to 70 yr of age in humans). In the current study, we observed an earlier release of recently dividing neutrophils from bone marrow and greater in-group variability of neutrophil kinetics based on in vivo BrdU labeling in a group of older rhesus macaques of 20-26 yr of age. Comparing neutrophil numbers and circulating cytokine levels in rhesus macaques spanning 2 to 26 yr of age, we found a negative correlation between age and blood neutrophil counts and a positive correlation between age and plasma G-CSF levels. Hierarchic clustering analysis also identified strong associations between G-CSF with the proinflammatory cytokines, IL-1β and MIP-1α. Furthermore, neutrophils from older macaques expressed less myeloperoxidase and comprised higher frequencies of polymorphonuclear myeloid-derived suppressor cells (PMN-MDSCs) compared to the young adult macaques. In summary, we observed an earlier release from bone marrow and a reduced production of neutrophils despite the increased levels of plasma G-CSF, especially in the elderly rhesus macaques. This lower neutrophil production capacity associated with increased production of proinflammatory cytokines as well as an earlier release of less mature neutrophils and PMN-MDSCs may contribute to the chronic inflammation and greater susceptibility to infectious and noninfectious diseases during aging.

Global health efforts have increased against infectious diseases, but issues persist with pathogens like Group B Streptococcus (GBS). Preclinical studies have elaborated on the mechanistic process of GBS-induced chorioamnionitis and its impact on the fetal programming of chronic neuropsychiatric diseases. GBS inoculation in rodents demonstrated the following: (i) silent and self-limited placental infection, similar to human chorioamnionitis; (ii) placental expression of chemokines attracting polymorphonuclear (PMN) cells; (iii) in vitro cytokine production; (iv) PMN infiltration in the placenta (histologic hallmark of human chorioamnionitis), linked to neurobehavioral impairments like cerebral palsy and autism spectrum disorders (ASD); (v) upregulation of interleukin-1β (IL-1β) in the placenta and fetal blood, associated with higher ASD risk in humans; (vi) sex-specific effects, with higher IL-1β release and PMN recruitment in male placenta; (vii) male offspring exhibiting ASD-like traits, while female offspring displayed attention deficit and hyperactivity disorder (ADHD)-like traits; (viii) IL-1 and/or NF-kB blockade alleviate placental and fetal inflammation, as well as subsequent neurobehavioral impairments. These findings offer potential therapeutic avenues, including sex-adapted anti-inflammatory treatment (e.g., blocking IL-1; repurposing of FDA-approved IL-1 receptor antagonist (IL-1Ra) treatment). Blocking the IL-1 pathway offers therapeutic potential to alleviate chorioamnionitis-related disabilities, presenting an opportunity for a human phase II RCT that uses IL-1 blockade added to the classic antibiotic treatment of chorioamnionitis. [ABSTRACT FROM AUTHOR]

Pathogen-associated molecular patterns (PAMPs) like bacterial cell wall components and viral nucleic acids are known ligands of innate inflammatory receptors that trigger multiple inflammatory pathways that may result in acute inflammation and oxidative stress-driven tissue and organ toxicity. When dysregulated, this inflammation may lead to acute toxicity and multiorgan failure. Inflammatory events are often driven by high energy demands and macromolecular biosynthesis. Therefore, we proposed that targeting the metabolism of lipopolysaccharide (LPS)-driven inflammatory events, using an energy restriction approach, can be an effective strategy to prevent the acute or chronic detrimental effects of accidental or seasonal bacterial and other pathogenic exposures. In the present study, we investigated the potential of energy restriction mimetic agent (ERMA) 2-deoxy-D-glucose (2-DG) in targeting the metabolism of inflammatory events during LPS-elicited acute inflammatory response. Mice fed with 2-DG as a dietary component in drinking water showed reduced LPS-driven inflammatory processes. Dietary 2-DG reduced LPS-induced lung endothelial damage and oxidative stress by strengthening the antioxidant defense system and limiting the activation and expression of inflammatory proteins, viz., P-Stat-3, NfκB, and MAP kinases. This was accompanied by decreased TNF, IL-1ß, and IL-6 levels in peripheral blood and bronchoalveolar lavage fluid (BALF). 2-DG also reduced the infiltration of PMNCs (polymorphonuclear cells) in inflamed tissues. Altered glycolysis and improved mitochondrial activity in 2-DG-treated RAW 264.7 macrophage cells suggested possible impairment of macrophage metabolism and, therefore, activation in macrophages. Taken together, the present study suggests that inclusion of glycolytic inhibitor 2-DG as a part of the diet can be helpful in preventing the severity and poor prognosis associated with inflammatory events during bacterial and other pathogenic exposures. [ABSTRACT FROM AUTHOR]

ABSTRACT: Endometritis is a uterine disease of dairy cows causing substantial negative effects on reproductive performance and inflicting considerable economic losses. It is typically diagnosed by endometrial cytology evaluation and commonly named cytological endometritis (CEM). In most previous studies, cows were defined as CEM positive if the proportion of polymorphonuclear cells (%PMN) in their endometrial cytology was above a pre-set threshold. Thresholds were established based on CEM diagnosis in association with reproductive performance, typically analyzed by a single reproductive parameter and calculated for all cows together. Our objective was to examine whether primiparous and multiparous cows should optimally be diagnosed for CEM by different %PMN thresholds and sampling timings, using a combination of several reproductive performance parameters. Two endometrial cytobrush cytology samples were collected from Holstein-Friesian dairy cows (n = 415; 269 multiparous; 146 primiparous), at 30–40 d in milk (DIM) and 60–70 DIM, and %PMN were evaluated microscopically (blindly; Diff-Quick stain, Medi-Market). The %PMN thresholds were set at ≥1% to ≥10%, ≥15%, and ≥20%, and accordingly, for each of the thresholds, several reproductive performance parameters were compared between CEM-positive versus CEM-negative cows. Upon application of several analytic approaches, our results indicated that optimal CEM diagnosis should be performed by different criteria in primiparous and multiparous cows: in primiparous cows at 30–40 DIM, using a threshold of ≥7%PMN, and in multiparous cows at 60–70 DIM, using a threshold of ≥4%PMN. Such a diagnostic approach provides a comprehensive view of the reproductive prognosis of CEM-positive primiparous and multiparous cows, which is pertinent information for researchers, veterinarians, and farmers.

Centella asiatica (C. asiatica) has reported to be one of the traditional herbal remedies, whereas poor water solubility leads to lower bioavailability thereby affecting it remedial efficacy. Therefore, we aimed to evaluate its efficacy through increased bioavailability by using high viscosity Carboxymethyl Cellulose (CMC) as solvent on methanol-based extract on wound healing, in vivo. The preparation was applied as 0.0% (control, CMC alone), 0.25. 0.5 and 1% concentrations of extract of C. asiatica. We evaluated the efficiency of preparations on wound healing progression as progression of wound contraction, tissue proliferation and cells deposition, and relative level of gene expression for genes associated with wound healing. The results showed that 0.5% extract in CMC had significantly higher (P < 0.05) wound contraction than control and other concentrations. The level tissue deposition and the infiltration of polymorphonuclear cells in groups treated with 0.5 % concentration preparation were higher than that other treatments and control. Similarly, the relative level of gene expression in 0.5% concentration treated group were statistically significantly higher (P < 0.05) than that of control. It is believed that the lower concentration of the extract would have lessor effect on wound healing, whereas higher concertation would be interfering the optimal inflammatory tissue deposition; and there by negatively affecting wound healing. The results indicated that C. asiatica can be optimally used at 0.5 % of extract in CMC for wound healing as indicated by speeding the progression of wound closure and by increasing the expression of collagen II and III together with reducing the expression of TGFβ1. However, higher concentrations of the crude extract of C. asiatica could paradoxically resulting in undesired effects. It is recommended that further evaluation should be performed on wider scale and the economic feasibility evaluation should be performed.

Abstract Background Ivermectin is widely used in human and animal medicine to treat and prevent parasite nematode infections. It has been suggested that its mode of action requires the host immune system, as it is difficult to reproduce its clinical efficacy in vitro. We therefore studied the effects of a single dose of ivermectin (Stromectol®—0.15 mg/kg) on cytokine levels and immune cell gene expression in human volunteers. This dose reduces bloodstream microfilariae rapidly and for several months when given in mass drug administration programmes. Methods Healthy volunteers with no travel history to endemic regions were given 3–4 tablets, depending on their weight, of either ivermectin or a placebo. Blood samples were drawn immediately prior to administration, 4 h and 24 h afterwards, and complete blood counts performed. Serum levels of 41 cytokines and chemokines were measured using Luminex® and expression levels of 770 myeloid-cell-related genes determined using the NanoString nCounter®. Cytokine levels at 4 h and 24 h post-treatment were compared to the levels pre-treatment using simple t tests to determine if any individual results required further investigation, taking p =

Endometritis is a uterine disease of dairy cows causing substantial negative effects on reproductive performance and inflicting considerable economic losses. It is typically diagnosed by endometrial cytology evaluation and commonly named cytological endometritis (CEM). In most previous studies, cows were defined as CEM positive if the proportion of polymorphonuclear cells (%PMN) in their endometrial cytology was above a pre-set threshold. Thresholds were established based on CEM diagnosis in association with reproductive performance, typically analyzed by a single reproductive parameter and calculated for all cows together. Our objective was to examine whether primiparous and multiparous cows should optimally be diagnosed for CEM by different %PMN thresholds and sampling timings, using a combination of several reproductive performance parameters. Two endometrial cytobrush cytology samples were collected from Holstein-Friesian dairy cows (n = 415; 269 multiparous; 146 primiparous), at 30–40 d in milk (DIM) and 60–70 DIM, and %PMN were evaluated microscopically (blindly; Diff-Quick stain, Medi-Market). The %PMN thresholds were set at ≥1% to ≥10%, ≥15%, and ≥20%, and accordingly, for each of the thresholds, several reproductive performance parameters were compared between CEM-positive versus CEM-negative cows. Upon application of several analytic approaches, our results indicated that optimal CEM diagnosis should be performed by different criteria in primiparous and multiparous cows: in primiparous cows at 30–40 DIM, using a threshold of ≥7%PMN, and in multiparous cows at 60–70 DIM, using a threshold of ≥4%PMN. Such a diagnostic approach provides a comprehensive view of the reproductive prognosis of CEM-positive primiparous and multiparous cows, which is pertinent information for researchers, veterinarians, and farmers. [ABSTRACT FROM AUTHOR]

Background: Ivermectin is widely used in human and animal medicine to treat and prevent parasite nematode infections. It has been suggested that its mode of action requires the host immune system, as it is difficult to reproduce its clinical efficacy in vitro. We therefore studied the effects of a single dose of ivermectin (Stromectol®—0.15 mg/kg) on cytokine levels and immune cell gene expression in human volunteers. This dose reduces bloodstream microfilariae rapidly and for several months when given in mass drug administration programmes. Methods: Healthy volunteers with no travel history to endemic regions were given 3–4 tablets, depending on their weight, of either ivermectin or a placebo. Blood samples were drawn immediately prior to administration, 4 h and 24 h afterwards, and complete blood counts performed. Serum levels of 41 cytokines and chemokines were measured using Luminex® and expression levels of 770 myeloid-cell-related genes determined using the NanoString nCounter®. Cytokine levels at 4 h and 24 h post-treatment were compared to the levels pre-treatment using simple t tests to determine if any individual results required further investigation, taking p = < 0.05 as the level of significance. NanoString data were analysed on the proprietary software, nSolver™. Results: No significant differences were observed in complete blood counts or cytokine levels at either time point between people given ivermectin versus placebo. Only three genes showed a significant change in expression in peripheral blood mononuclear cells 4 h after ivermectin was given; there were no significant changes 24 h after drug administration or in polymorphonuclear cells at either time point. Leukocytes isolated from those participants given ivermectin showed no difference in their ability to kill Brugia malayi microfilariae in vitro. Conclusions: Overall, our data do not support a direct effect of ivermectin, when given at the dose used in current filarial elimination programmes, on the human immune system. Trial registration ClinicalTrials.gov NCT03459794 Registered 9th March 2018, Retrospectively registered https://clinicaltrials.gov/ct2/show/NCT03459794?term=NCT03459794&draw=2&rank=1. [ABSTRACT FROM AUTHOR]

Background: Leishmaniasis is a neglected arthropod-borne disease that affects millions of people worldwide. Successful Leishmania infections require the mitigation of immune cell functions leading to parasite survival and proliferation. A large body of evidence highlights the involvement of neutrophils (PMNs) and dendritic cells (DCs) in the establishment of immunological responses against these parasites. However, few studies, contemplate to what extent these cells interact synergistically to constrain Leishmania infection. Objective: We sought to investigate how PMNs and infected DCs interact in an in vitro model of Leishmania amazonensis infection. Material and Methods: Briefly, human PMNs and DCs were purified from the peripheral blood of healthy donors. Next, PMNs were activated with fibronectin and subsequently co-cultured with L. amazonensis -infected DCs. Results: We observed that L. amazonensis -infected DC exhibited lower rates of infection when co-cultivated with either resting or activated PMNs. Surprisingly, we found that the release of neutrophil enzymes was not involved in Leishmania killing. Next, we showed that the interaction between PMNs and infected-DCs was intermediated by DC-SIGN, further suggesting that parasite elimination occurs in a contact-dependent manner. Furthermore, we also observed that TNFα and ROS production was dependent on DC-SIGN-mediated contact, as well as parasite elimination is dependent on TNFα production in the co-culture. Finally, we observed that direct contact between PMNs and DCs are required to restore the expression of DC maturation molecules during L. amazonensis infection. Conclusion: Our findings suggest that the engagement of direct contact between PMNs and L. amazonensis -infected DC via DC-SIGN is required for the production of inflammatory mediators with subsequent parasite elimination and DC maturation. [ABSTRACT FROM AUTHOR]

BackgroundLeishmaniasis is a neglected arthropod-borne disease that affects millions of people worldwide. Successful Leishmania infections require the mitigation of immune cell functions leading to parasite survival and proliferation. A large body of evidence highlights the involvement of neutrophils (PMNs) and dendritic cells (DCs) in the establishment of immunological responses against these parasites. However, few studies, contemplate to what extent these cells interact synergistically to constrain Leishmania infection.ObjectiveWe sought to investigate how PMNs and infected DCs interact in an in vitro model of Leishmania amazonensis infection.Material and MethodsBriefly, human PMNs and DCs were purified from the peripheral blood of healthy donors. Next, PMNs were activated with fibronectin and subsequently co-cultured with L. amazonensis-infected DCs.ResultsWe observed that L. amazonensis-infected DC exhibited lower rates of infection when co-cultivated with either resting or activated PMNs. Surprisingly, we found that the release of neutrophil enzymes was not involved in Leishmania killing. Next, we showed that the interaction between PMNs and infected-DCs was intermediated by DC-SIGN, further suggesting that parasite elimination occurs in a contact-dependent manner. Furthermore, we also observed that TNFα and ROS production was dependent on DC-SIGN-mediated contact, as well as parasite elimination is dependent on TNFα production in the co-culture. Finally, we observed that direct contact between PMNs and DCs are required to restore the expression of DC maturation molecules during L. amazonensis infection.ConclusionOur findings suggest that the engagement of direct contact between PMNs and L. amazonensis-infected DC via DC-SIGN is required for the production of inflammatory mediators with subsequent parasite elimination and DC maturation.

Background: It is commonly believed that cancer development is irreversible, organ-specific as well as systemic malignant disorder, often associated with harmful oxidative stress and inflammation. However, there are also well-documented cases of spontaneous cancer regression, the causative mechanisms of which are not understood. It is known that inflammation is a negative pathophysiological process that may support the development of cancer, but it is also believed that the immune system as well as oxidative stress play important roles in prevention of cancer development and defense against tumor progression. Hence, in animal models spontaneous regression of cancer could be mediated by rapid inflammatory response of granulocytes, acting against cancer mostly as innate immune response. In addition, the administration of granulocytes at the site of solid tumors can lead to tumor regression or can slow down tumor growth and extend the overall survival of animals. In both cases, similar to the radiotherapy, surgery and various chemotherapies, oxidative stress occurs generating lipid peroxidation product 4-hydroxynonenal (4-HNE). This “second messenger of free radicals” acts as growth regulating signaling molecule that exerts relatively selective cytotoxicity against cancer cells. Conclusions: We hypothesize that abundant inflammation and metabolic changes caused by cancer and oxidative stress producing of 4-HNE may be crucial mechanisms for spontaneous cancer regression.

Abstract Background The human skin is an extremely sophisticated and evolved organ that covers the whole body. External agents or the patient’s own diseases can cause skin injuries that can challenge healthcare professionals and impose high social, economic and emotional costs. Objectives To evaluate the impact of topical nifedipine on skin wound healing, specifically on polymorphonuclear cells, vascular proliferation, and collagen. Methods We used three pigs, and created eight injuries in the dorsal region of each animal. We applied 1%, 10%, and 20% concentration nifedipine creams to four of the wounds in animals 1, 2, and 3 respectively and treated the other twelve wounds with saline solution 0.9% only. We analyzed the presence of polymorphonuclear cells, vascular proliferation, and collagen at six different times (days 1, 3, 7, 14, 21, and 28). Results The evaluation of polymorphonuclear levels showed mild cell activity at all times in the control group, while in the nifedipine groups, marked levels were more frequent at all times during the experiment. There was a 4.84-fold increase in the chance of marked vascular proliferation (p = 0.019) and, at the same time, a decrease in collagen formation (OR 0.02 / p = 0.005) in animal 3. Conclusions Topical NFD may have an impact on skin wound healing mechanisms. Our study showed that polymorphonuclear cells and vascular proliferation increased. We also demonstrated that collagen formation decreased. Therefore, topical NFD may have a positive impact on skin wound healing. Additional studies are needed to confirm our results.

Mycoplasma mycoides subsp. mycoides (Mmm) is the etiological agent of contagious bovine pleuropneumonia (CBPP), one of the major diseases affecting cattle in sub-Saharan Africa. Some evidences suggest that the immune system of the host (cattle) plays an important role in the pathogenic mechanism of CBPP, but the factors involved in the process remain largely unknown. The present study aimed to investigate the cell response of bovine polymorphonuclear neutrophils (PMNs) after Mmm in vitro exposure using one step RT-qPCR and Western blotting. Data obtained indicate that gene and protein expression levels of some pro-inflammatory factors already change upon 30 min of PMNs exposure to Mmm. Of note, mRNA expression level in Mmm exposed PMNs increased in a time-dependent manner and for all time points investigated; targets expression was also detected by Western blotting in Mmm exposed PMNs only. These data demonstrate that when bovine PMN cells are triggered by Mmm, they undergo molecular changes, upregulating mRNA and protein expression of specific pro-inflammatory factors. These results provide additional information on host-pathogen interaction during CBPP infection.

Uterine epithelial cells (UEC) and migrated polymorphonuclear cells (PMN) play important roles in the uterine defence against microbial infection. The aims of the present study were to investigate i) whether undiluted uterine secretions modulate the expression of genes associated with the innate immune response in UEC and PMN in vitro , ii) whether these changes differ between the two cell populations and iii) whether uterine secretions from cows with subclinical endometritis produce a different response to those from unaffected cows. Therefore, undiluted uterine secretions, cytobrush and biopsy samples were collected from bovine uteri at a local abattoir. All cows had calved at least 3 months prior to sample collection. Subclinical endometritis was diagnosed by cytology (≥5% polymorphonuclear neutrophils) and histology. The uteri were thereby retrospectively categorised as endometritis-positive (E-pos; n = 14), if either the cytology or the histology results were positive, or endometritis-negative (E-neg; n = 17), if both diagnostics were negative. Cultured UEC responded to secretions from E-pos and E-neg cows with an increased gene expression of CXC ligand (CXCL) 8 and interleukin (IL) 6 compared to incubation with control medium alone. PMN expressed significantly higher mRNA levels of CXCL5, CXCL8 and IL1B in response to supernatant from UEC incubated with secretions from both groups (E-pos and E-neg) compared to those incubated with control medium alone. Gene expression of IL10 in uterine epithelial cells remained comparable to the control in cells exposed to E-pos secretions and was 3.6 times lower in those exposed to E-neg secretions. These results demonstrate that the expression of genes associated with the innate immune response in UEC and indirectly also PMN is affected by uterine secretions in vitro. Depending on the target gene, these changes differ between the two cell populations. UEC exposed to uterine secretions from cows without subclinical endometritis produce lower levels of IL10 compared to those exposed to secretions from affected cows or control medium alone. Therefore, the model established in this study can be used as a valuable tool to further understand the contributions of the two cell populations to the coordinated immune response in the uterus. • Bovine uterine secretions can be collected post mortem without dilution by flushing media. • Undiluted uterine secretions can modulate the expression of immune genes in UEC. • Supernatant from UEC stimulated by uterine secretions can influence gene expression in PMN. • Secretions from cows with subclinical endometritis change the response in the 2 cell types. [ABSTRACT FROM AUTHOR]

Brucellosis is a contagious disease caused by bacteria of the genus Brucella. Platelets (PLTs) have been widely involved in the modulation of the immune response. We have previously reported the modulation of Brucella abortus–mediated infection of monocytes. As a result, PLTs cooperate with monocytes and increase their inflammatory capacity, promoting the resolution of the infection. Extending these results, in this study we demonstrate that patients with brucellosis present slightly elevated levels of complexes between PLTs and both monocytes and neutrophils. We then assessed whether PLTs were capable of modulating functional aspects of neutrophils. The presence of PLTs throughout neutrophil infection increased the production of interleukin‐8, CD11b surface expression and reactive oxygen species formation, whereas it decreased the expression of CD62L, indicating an activated status of these cells. We next analyzed whether this modulation was mediated by released factors. To discriminate between these options, neutrophils were treated with supernatants collected from B. abortus–infected PLTs. Our results show that CD11b expression was induced by soluble factors of PLTs but direct contact between cell populations was needed to enhance the respiratory burst. Additionally, B. abortus–infected PLTs recruit polymorphonuclear (PMN) cells to the site of infection. Finally, the presence of PLTs did not modify the initial invasion of PMN cells by B. abortus but improved the control of the infection at extended times. Altogether, our results demonstrate that PLTs interact with neutrophils and promote a proinflammatory phenotype which could also contribute to the resolution of the infection. [ABSTRACT FROM AUTHOR]

Background: The human skin is an extremely sophisticated and evolved organ that covers the whole body. External agents or the patient's own diseases can cause skin injuries that can challenge healthcare professionals and impose high social, economic and emotional costs. Objectives: To evaluate the impact of topical nifedipine on skin wound healing, specifically on polymorphonuclear cells, vascular proliferation, and collagen. Methods: We used three pigs, and created eight injuries in the dorsal region of each animal. We applied 1%, 10%, and 20% concentration nifedipine creams to four of the wounds in animals 1, 2, and 3 respectively and treated the other twelve wounds with saline solution 0.9% only. We analyzed the presence of polymorphonuclear cells, vascular proliferation, and collagen at six different times (days 1, 3, 7, 14, 21, and 28). Results: The evaluation of polymorphonuclear levels showed mild cell activity at all times in the control group, while in the nifedipine groups, marked levels were more frequent at all times during the experiment. There was a 4.84-fold increase in the chance of marked vascular proliferation (p = 0.019) and, at the same time, a decrease in collagen formation (OR 0.02 / p = 0.005) in animal 3. Conclusions: Topical NFD may have an impact on skin wound healing mechanisms. Our study showed that polymorphonuclear cells and vascular proliferation increased. We also demonstrated that collagen formation decreased. Therefore, topical NFD may have a positive impact on skin wound healing. Additional studies are needed to confirm our results. [ABSTRACT FROM AUTHOR]

The human and animal pathogens Yersinia pestis, which causes bubonic and pneumonic plague, and Yersinia pseudotuberculosis and Yersinia enterocolitica, which cause gastroenteritis, share a type 3 secretion system which injects effector proteins, Yops, into host cells. This system is critical for virulence of all three pathogens in tissue infection. Neutrophils are rapidly recruited to infected sites and all three pathogens frequently interact with and inject Yops into these cells during tissue infection. Host receptors, serum factors, and bacterial adhesins appear to collaborate to promote neutrophil– Yersinia interactions in tissues. The ability of neutrophils to control infection is mixed depending on the stage of infection and points to the efficiency of Yops and other bacterial factors to mitigate bactericidal effects of neutrophils. Yersinia in close proximity to neutrophils has higher levels of expression from yop promoters, and neutrophils in close proximity to Yersinia express higher levels of pro-survival genes than migrating neutrophils. In infected tissues, YopM increases neutrophil survival and YopH targets a SKAP2/SLP-76 signal transduction pathway. Yet the full impact of these and other Yops and other Yersinia factors on neutrophils in infected tissues has yet to be understood.

Background: Endometritis, which is one of the most common diseases in dairy cows postpartum, causes severe economic losses, including increased open days, calving intervals, and numbers of services to achieve conception. Aim: This study aimed to evaluate the ultrasound method and its agreement with the endometrium cytology method, which is used to diagnose cytological endometritis in beef cows. Moreover, we determined which method has higher sensitivity and specificity at 4 and 5 weeks postpartum. Materials and Methods: The study was conducted 20-35 days postpartum. A total of 53 clinically healthy beef cows (28 Brangus and 25 Kedah-Kelantan breeds) from three beef farms were obtained. All cows were evaluated at 4 and 5 weeks postpartum, using ultrasound and cytobrush endometrial examination methods to diagnose cytological endometritis. Results: Endometrial cytology result showed that 11.3% (6/53) and 9.4% (5/53) of the cows exhibited cytological endometritis 4 and 5 weeks postpartum, respectively. A weak-to-moderate agreement found between the diagnostic methods (k=0.29 - 0.50; p

Neutrophils play a central role in immunity and inflammation via their intrinsic ability to migrate into inflamed tissue, to phagocytose pathogens, and to kill bacterial and fungi by releasing large quantities of superoxide anions and lytic enzymes. The molecular pathways controlling neutrophil microbicidal functions are still unclear, because neutrophils have a short half‐life and are resistant to genetic manipulation. Neutrophil‐like cells (NLC) can be generated from myeloid progenitors conditionally immortalized with the ER‐HoxB8 oncoprotein, but whether these cells can replace neutrophils in high‐throughput functional assays is unclear. Here, we assess the ability of NLC derived from ER‐HoxB8 progenitors to produce ROS and to perform chemotaxis and phagocytosis. We compare the Ca2+ responses and effector functions of NLC to primary murine neutrophils and document the molecular basis of their functional differences by mRNA profiling. Pro‐inflammatory cytokines enhanced the expression by NLC of neutrophil surface markers and transcription factors. Ca2+ elevations evoked in NLC by agonists, adhesion receptors, and store depletion resembled the physiological responses recorded in primary neutrophils, but NLC expressed reduced amounts of Ca2+ signaling proteins and of chemotactic receptors. Unlike their myeloid progenitors, NLC produced H2O2 when adhered to fibronectin, migrated toward chemotactic peptides, phagocytosed opsonized particles, and generated intracellular ROS. NLC phagocytosed as efficiently as primary neutrophils but produced 50 times less ROS and migrated less efficiently toward chemoattractant. Our data indicate that NLC can replace neutrophils to study Ca2+ signaling and phagocytosis, but that their incomplete granulocytic differentiation limits their use for chemotaxis and ROS production assays. [ABSTRACT FROM AUTHOR]

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Background: Neonatal pneumonia (NP) is one of the major causes of neonatal death. Current NP diagnosis depends on a detailed history, physical examination, and radiographic and laboratory findings. There is no specific biomarker or diagnostic indicator of NP. Methods: In this study, we tried to find a reliable biomarker for quick NP diagnosis by collecting peripheral blood from neonates with NP and transient tachypnea of the newborn (TTN), and subsequently tested the expression of CD64 on white blood cells using flow cytometry. The cellularity of each blood cell population was also quantified. Furthermore, procalcitonin (PCT) and C-reactive protein (CRP) levels were evaluated in the blood sera. Results: We found that NP patients had moderately increased polymorphonuclear cells (PMNs), as well as elevated PCT and CRP levels in the blood sera. Importantly, the expression of CD64 on PMNs was profoundly increased in NP patients but not TTN patients. The receiver operating characteristic (ROC) curve of PMN CD64 index suggests that PMN CD64 index is sensitive and specific for NP diagnosis. Conclusions: Our study reveals that PMN CD64 could be a fast and reliable biomarker for NP diagnosis. [ABSTRACT FROM AUTHOR]

Aim: Poly(ε-caprolactone) lipid-core nanocapsules (LNCs) are efficient drug carriers and drug-free LNCs display therapeutic effects, inhibiting tumor growth and neutrophil activities. Herein, we investigated the direct actions of LNCs on human immune cells, to guide their therapeutic application. Materials & methods: LNC's uptake, cytokine release, cell migration, proliferation and intracellular pathways under inflammatory stimulation were investigated. Results & conclusion: LNCs quickly penetrated leukocytes without cytotoxicity; inhibited mitogen-induced lymphocyte proliferation, cytokine release and leukocyte migration under inflammatory stimulation, which were associated with inhibition of the MAP kinase pathway and intracellular calcium influx. Hence, we showed LNCs as a down-regulatory agent on immune cells, suggesting that either the particles themselves or their application as a drug carrier can halt non-desired inflammatory processes. [ABSTRACT FROM AUTHOR]

Recent studies have shown that melatonin can be secreted, in addition to other sources, by peripheral blood mononuclear cells (PBMCs). We investigated whether melatonin could also be secreted by cultured polymorphonuclear (PMNs) and whole blood (WB) cells under resting and stimulated conditions. In addition, influences of the intensity and color of light on the production of melatonin by these cells were estimated. Blood samples were collected from a healthy volunteer, and PBMCs and PMNs were isolated. The PBMC, PMN, and WB cultures were exposed for 8 h to light intensities of 0, 20, 50, 100, 300, and 500 lux and to white, red, blue, and green lights in the presence/absence of mitogens. Levels of melatonin in supernatants were detected by enzyme-linked immunosorbent assay (ELISA). In addition to PBMCs, PMNs and WB cells were found to also produce melatonin, and this production was increased due to mitogen activations. The most effective intensity of light, which suppresses melatonin secretion, was 100 lux. There were no significant differences between the influences of different monochromatic lights on the melatonin production by the examined blood cell cultures. Thus, PBMCs, PMNs, and WB cultures can produce melatonin, and mitogen activation causes a higher production of this hormone. Moreover, although white light suppressed melatonin secretion at 100 lux, there was no difference among various types of lights. [ABSTRACT FROM AUTHOR]

Paraquat poisoning and hyperbaric oxygen exposure are well established models of oxidative stress in lung. The aim of this study was focused on the contribution of oxygen free radicals and other cytotoxic species, such as lipid hydroperoxides, to the overall toxicity. Adult Wistar rats were injected with paraquat (30 or 60 mg/kg b.w.) or exposed to hyperbaric oxygen (0.2 MPa), and several parameters of lung damage were measured. Both treatments resulted in increased spontaneous lung chemiluminescence, number of lung PMN, malondialdehyde content, lung edema, and pleural liquid. Of note, spontaneous lung chemiluminescence, used to monitor the steady-state concentration of oxygen free radicals in vivo, did not increase significantly after either treatment. The increase in spontaneous lung chemiluminescence started after PMN migration, being both maxima separated by a delay time of 4-6 h. After PMN migrated and became activated in the lung, the survival of the animals started to decline. Thus, PMN can be considered as additional sources of oxygen free radicals supported by the subsequent increase in chemiluminescence. Their role in lung damage was evidenced by the increase in lung edema, augmented pleural liquid, and decreased survival after PMN migration. Lipid hydroperoxide concentration in lung membranes was also increased after either treatment. This increased concentration may be a consequence of an increased rate of lipid peroxidation, initiated by oxidative stress on lipid membranes, or by an inhibition of their catabolism. Ester lipid hydroperoxides normally produced in membranes cannot be catabolized directly by the glutathione peroxidase-reductase system unless phospholipase A2 catalyses the release of free lipid hydroperoxides. In both experimental models, phospholipase A2 activity was decreased to almost negligible values. Betamethasone (1 mg/ml; IV) administered to the rats 3 h before paraquat injection accelerated the decrease in survival and phospholipase A2 inactivation. Inactivation of phospholipase A2, detected in paraquat or oxygen exposed rats, could be attributed to a O2(.-)-driven Fenton reaction. However, phospholipase A2 inactivation by betamethasone pretreatment may be attributed to the presence of lipocortin, a corticosteroid-inducible factor and inhibitor of phospholipase A2. Besides the mechanism underlying the inactivation of phospholipase A2, the increase in lipid hydroperoxides may indicate their role as long-lived cytotoxic species that contribute to the damage already initiated by oxidative stress. Indeed, lipid hydroperoxides are very well known modifiers of membrane physical properties.

Context: Tobacco smoking is considered to be a major risk factor associated with periodontal disease. Smoking exerts a major effect on the protective elements of the immune response, resulting in an increase in the extent and severity of periodontal destruction. Aims: The aim of the present study was to assess viability and phagocytic function of neutrophils in circulating blood of the smokers and nonsmokers who are periodontally healthy. Settings and Design: Two hundred subjects in the mean range of 20–30 years of age were included in the study population. It was a retrospective study carried out for 6 months. Materials and Methods: Two hundred subjects were divided into four groups: 50 nonsmokers, 50 light smokers (15 cigarettes/day). Full mouth plaque index, sulcus bleeding index, and probing depths were measured. Percentage viability of circulating neutrophils and average number of phagocytosed Candida albicans were recorded. Statistical Analysis Used: Means and standard deviations were calculated from data obtained within the groups. Comparison between the smokers and nonsmokers was performed by Kruskal–Wallis ANOVA analysis. Comparison between smoker groups was performed using Mann–Whitney–Wilcoxon test. Results: Percentage viability of neutrophils was significantly less in heavy smokers (66.9 ± 4.0), moderate (76.6 ± 4.2), light smokers (83.1 ± 2.5) as compared to nonsmokers (92.3 ± 2.6) (P < 0.01). The ability of neutrophils to phagocytose, i.e., mean particle number was significantly less in light smokers (3.5 ± 0.5), moderate smokers (2.3 ± 0.5), and heavy smokers (1.4 ± 0.5) compared to nonsmokers (4.9 ± 0.7) (P < 0.01) with evidence of dose-response effect. Conclusions: Smoking significantly affects neutrophils viability and phagocytic function in periodontally healthy population.

A study was conducted in 24 crossbred cows and endometrial sampling was performed by uterine cytobrush (UC) on 30, 40 and 50 days postpartum (DPP) to diagnose subclinical endometritis (SCE). Out of which 10 cows were diagnosed to be positive for SCE and were treated with E.colilipopolysaccharide (LPS). Polymorphonuclear (PMN) cells count byUC showed a significant reduction (P

Introduction: Joint effusion is a common clinical problem resulting from various inflammatory and non-inflammatory conditions. Synovial fluid analysis is one of the most important diagnostic tests to classify the type of effusion. Aim: To assess the role of synovial fluid analysis in the diagnosis of joint diseases and also to assess the pattern of arthritis in tertiary care centre. Materials and Methods: A total of 100 samples of synovial fluid were examined. Synovial fluids were subjected to gross examination for colour, viscosity, appearance, wet mount preparation, total and differential leukocyte count, microbiologic assays and crystal analysis by polarised microscopy. Results: Out of 100 samples analysed, Effusion was noninflammatory in 32 (32%) cases, inflammatory in 46 (46%) cases, infective in 10 (10%) cases, traumatic in 3 (3%) cases, crystal induced in 3 (3%) cases, normal in 4 (4%) cases and non-diagnostic in 2 (2%) cases. The majority of cases were of osteoarthritis 22 (22%), then rheumatoid arthritis 16 (16%), tuberculous arthritis in 6 (6%) cases, septic arthritis in 4 (4%) cases, gout in 3 (3%) cases and traumatic arthritis in 3 (3%) cases. 10 (10%) cases were of non-inflammatory arthritis-not otherwise specified and 30 (30%) cases were of inflammatory arthritis-not otherwise specified. A total of 4 (4%) cases were normal and 2 (2%) cases were non-diagnostic. Total leukocyte count was found to be highest in septic arthtritis (60,000-78,000 cells/mm3 ) and lowest in osteoarthritis (120-23,00 cells/mm3 ). Polymorphs were highest in septic arthtritis (95%) and lowest in non-inflammatory conditions like osteoarthritis (23%). Sensitivity and specificity for gross examination to differentiate between inflammatory and non-inflammatory conditions was 0.91 and 0.70 respectively while it was 0.94 and 0.86 respectively for white blood cell count. Conclusion: Synovial fluid analysis is a simple procedure. The study of synovial fluid is a tool that allows the establishment of diagnosis and treatment of patient with joint diseases. Gross examination, total leukocyte count and differential leukocyte count accomplish well to differentiate between various inflammatory and non-inflammatory arthritis and also to diagnose septic arthritis.

Polymorphisms impacting thymic function may decrease peripheral tolerance and hasten autoimmune disease. The NF-κB transcription factor subunit, RelB, is essential for the development and differentiation of medullary thymic epithelial cells (mTECs): RelB-deficient mice have reduced thymic cellularity and markedly fewer mTECs, lacking AIRE. The precise mechanism of this mTEC reduction in the absence of RelB is unclear. To address this, we studied mTECs and dendritic cells (DCs), which critically regulate negative selection, and thymic regulatory T-cells (tTreg) in RelB−/− mice, which have spontaneous multiorgan autoimmune disease. RelB−/− thymi were organized, with medullary structures containing AIRE− mTECs, DCs, and CD4+ thymocytes, but fewer tTreg. Granulocytes infiltrated the RelB−/− thymic cortex, capsule, and medulla, producing inflammatory thymic medullary atrophy, which could be treated by granulocyte depletion or RelB+ DC immunotherapy, with concomitant recovery of mTEC and tTreg numbers. These data indicate that central tolerance defects may be accelerated by autoimmune thymic inflammation where impaired RelB signaling impairs the medullary niche, and may be reversible by therapies enhancing peripheral Treg or suppressing inflammation.

Introduction: Pro-inflammatory mediators are mainly responsible for periodontal tissue breakdown in periodontitis. Ceruloplasmin is one such biomarker, the levels of which seems to be elevated in presence of inflammation. Hence, assessing biomarkers of inflammation is a valuable tool for determining periodontal disease activity. Aim: To evaluate and compare the levels of serum ceruloplasmin in chronic generalised gingivitis, chronic generalised periodontitis and generalized aggressive periodontitis patients. Materials and Methods: Patients visiting the dental Outpatient Department (OPD) during the period of January 2017 to March 2017 were screened for gingival index, probing pocket depth, bleeding on probing and categorised into the following four groups based on the clinical signs and symptoms; healthy controls, chronic generalised gingivitis, chronic generalised periodontitis and generalised aggressive periodontitis groups. Ten patients were enrolled in each group. Blood samples were collected from the antecubital vein of the subjects. The collected samples were taken for serum ceruloplasmin assessment with essential precautions. The test results were subjected to statistical analysis using Tukey’s multiple post-hoc procedure, Karl pearson’s correlation coefficient and One-way ANOVA. results: The results showed that serum ceruloplasmin level in chronic periodontitis was increased when compared to gingivitis group. Further, rise in levels of serum ceruloplasmin was comparatively higher in aggressive periodontitis subjects than chronic periodontitis subjects. conclusion: Inflammatory markers are being used extensively for the diagnosis of periodontal diseases. It can be inferred from the results of the present study that ceruloplasmin is one such biomarker which seems to be elevated with progress in attachment loss.

Hyperammonaemia is a common complication of liver disease in dogs. High concentrations of ammonia can be detrimental to dogs with liver disease for several reasons, notably by causing hepatic encephalopathy (HE) which describes the wide range of neurological abnormalities ranging from altered behaviour to seizures that are well recognised complications in dogs with hepatic disorders. In human patients with liver disease, hyperammonaemia has also been linked to the development of other systemic complications such as dysregulation of the innate immune system. In contrast, the effects of hyperammonaemia on the canine innate immune system is currently unknown. The aim of this study was to investigate the effects of ammonia on the oxidative burst activity of canine polymorphonuclear cells in vitro. Blood obtained from healthy dogs (n = 8) was incubated with escalating concentrations of ammonia ranging from 0 to 250 μM, and the percentage of cells experiencing an oxidative burst was evaluated using a commercial kit (Phagoburst™) and flow cytometry. The spontaneous oxidative burst was evaluated without stimulation and also following stimulation with E coli. The pH of the blood was also measured at the differing ammonia concentrations. There was an increase in the percentage of cells experiencing a spontaneous oxidative burst from ammonia concentrations of 125 μM (p = <0.05) and above (p = <0.01), with a 4.9 fold increase at 200 μM (p = < 0.001). In those cells stimulated with E coli, incubation with increasing ammonia concentrations did not result in a significant difference in oxidative burst from baseline (p = 0.953). There was no statistically significant difference between the pH of the blood at the various ammonia concentrations (p = 0.2) suggesting that the difference in spontaneous oxidative burst was due to the ammonia rather than simply a change in pH conditions. In summary, the spontaneous oxidative burst of neutrophils was significantly increased from baseline. This supports a potential role of ammonia in contributing to innate immune system dysfunction in dogs with liver disease, and may present a future therapeutic target. [ABSTRACT FROM AUTHOR]

Reactive oxygen species (ROS) are suggested to play a role in the pathogenesis of contagious bovine pleuropneumonia, a severe respiratory disorder caused by Mycoplasma mycoides subsp. mycoides ( Mmm ). The present study investigated the generation of ROS by different strains of Mmm , as well as their effect on the oxidative response of bovine neutrophils. The production of ROS was indirectly measured using a luminol-based chemiluminescence assay. Our results confirm that Mmm can produce ROS via the metabolism of glycerol, significant differences existing between African and European strains. Mmm was capable of adhering to the external surface of neutrophils. Interestingly, Mmm enhanced the respiratory burst of bovine neutrophils. This activity was particularly pronounced with the African field strain and in presence of glycerol. Taken together, our data argue in favour of a major role for neutrophils as the main source of ROS in contagious bovine pleuropneumonia. [ABSTRACT FROM AUTHOR]

Introduction: Joint effusion is a common clinical problem resulting from various inflammatory and non-inflammatory conditions. Synovial fluid analysis is one of the most important diagnostic tests to classify the type of effusion. Aim: To assess the role of synovial fluid analysis in the diagnosis of joint diseases and also to assess the pattern of arthritis in tertiary care centre. Materials and Methods: A total of 100 samples of synovial fluid were examined. Synovial fluids were subjected to gross examination for colour, viscosity, appearance, wet mount preparation, total and differential leukocyte count, microbiologic assays and crystal analysis by polarised microscopy. Results: Out of 100 samples analysed, Effusion was noninflammatory in 32 (32%) cases, inflammatory in 46 (46%) cases, infective in 10 (10%) cases, traumatic in 3 (3%) cases, crystal induced in 3 (3%) cases, normal in 4 (4%) cases and non-diagnostic in 2 (2%) cases. The majority of cases were of osteoarthritis 22 (22%), then rheumatoid arthritis 16 (16%), tuberculous arthritis in 6 (6%) cases, septic arthritis in 4 (4%) cases, gout in 3 (3%) cases and traumatic arthritis in 3 (3%) cases. 10 (10%) cases were of non-inflammatory arthritis-not otherwise specified and 30 (30%) cases were of inflammatory arthritis-not otherwise specified. A total of 4 (4%) cases were normal and 2 (2%) cases were non-diagnostic. Total leukocyte count was found to be highest in septic arthtritis (60,000-78,000 cells/mm3) and lowest in osteoarthritis (120-23,00 cells/mm3). Polymorphs were highest in septic arthtritis (95%) and lowest in non-inflammatory conditions like osteoarthritis (23%). Sensitivity and specificity for gross examination to differentiate between inflammatory and non-inflammatory conditions was 0.91 and 0.70 respectively while it was 0.94 and 0.86 respectively for white blood cell count. Conclusion: Synovial fluid analysis is a simple procedure. The study of synovial fluid is a tool that allows the establishment of diagnosis and treatment of patient with joint diseases. Gross examination, total leukocyte count and differential leukocyte count accomplish well to differentiate between various inflammatory and non-inflammatory arthritis and also to diagnose septic arthritis. [ABSTRACT FROM AUTHOR]

Polymorphisms impacting thymic function may decrease peripheral tolerance and hasten autoimmune disease. The NF-κB transcription factor subunit, RelB, is essential for the development and differentiation of medullary thymic epithelial cells (mTECs): RelB-deficient mice have reduced thymic cellularity and markedly fewer mTECs, lacking AIRE. The precise mechanism of this mTEC reduction in the absence of RelB is unclear. To address this, we studied mTECs and dendritic cells (DCs), which critically regulate negative selection, and thymic regulatory T-cells (tTreg) in RelB-/- mice, which have spontaneous multiorgan autoimmune disease. RelB-/- thymi were organized, with medullary structures containing AIRE-mTECs, DCs, and CD4+ thymocytes, but fewer tTreg. Granulocytes infiltrated the RelB-/- thymic cortex, capsule, and medulla, producing inflammatory thymic medullary atrophy, which could be treated by granulocyte depletion or RelB+ DC immunotherapy, with concomitant recovery of mTEC and tTreg numbers. These data indicate that central tolerance defects may be accelerated by autoimmune thymic inflammation where impaired RelB signaling impairs the medullary niche, and may be reversible by therapies enhancing peripheral Treg or suppressing inflammation. [ABSTRACT FROM AUTHOR]

Abstract: Proton‐pump inhibitors (PPI) as pantoprazole are highly effective acid suppressive agents that belong to the world's most sold medication. However, they are pronounced to have immunosuppressive aspects. In our study, a negative influence of PPI on functions of polymorphonuclear cells in vitro like phagocytosis, oxidative burst, chemotaxis, and killing activity was shown, whereas formation of neutrophil extracellular traps (NET)osis remained unaffected. Pantoprazole stimulation additionally reduced the production of the proinflammatory cytokine IL‐1β in whole blood assay as well as the production of IL‐2 and IFN‐γ after whole blood stimulation with phytohaemagglutinin. Moreover, IFN‐γ feedback mechanisms and signaling by STAT‐1 was impaired by PPI. Cardiac surgery is accompanied by developing systemic inflammatory response syndrome with immunosuppressive aspects. We exhibited reduced oxidative burst analyzing cardiac surgery patients’ samples receiving or not receiving PPI. Furthermore, a higher rate of infections in patients receiving permanent PPI medication in retrospective analysis was uncovered. Patients undergoing cardiac surgery with cardiopulmonary bypass and regular PPI medication developed significant more infections retrospectively indicating a clinical impact of the immunosuppressive influence of PPI. [ABSTRACT FROM AUTHOR]

Introduction: Pro-inflammatory mediators are mainly responsible for periodontal tissue breakdown in periodontitis. Ceruloplasmin is one such biomarker, the levels of which seems to be elevated in presence of inflammation. Hence, assessing biomarkers of inflammation is a valuable tool for determining periodontal disease activity. Aim: To evaluate and compare the levels of serum ceruloplasmin in chronic generalised gingivitis, chronic generalised periodontitis and generalized aggressive periodontitis patients. Materials and Methods: Patients visiting the dental Outpatient Department (OPD) during the period of January 2017 to March 2017 were screened for gingival index, probing pocket depth, bleeding on probing and categorised into the following four groups based on the clinical signs and symptoms; healthy controls, chronic generalised gingivitis, chronic generalised periodontitis and generalised aggressive periodontitis groups. Ten patients were enrolled in each group. Blood samples were collected from the antecubital vein of the subjects. The collected samples were taken for serum ceruloplasmin assessment with essential precautions. The test results were subjected to statistical analysis using Tukey’s multiple post-hoc procedure, Karl pearson’s correlation coefficient and One-way ANOVA. Results: The results showed that serum ceruloplasmin level in chronic periodontitis was increased when compared to gingivitis group. Further, rise in levels of serum ceruloplasmin was comparatively higher in aggressive periodontitis subjects than chronic periodontitis subjects. Conclusion: Inflammatory markers are being used extensively for the diagnosis of periodontal diseases. It can be inferred from the results of the present study that ceruloplasmin is one such biomarker which seems to be elevated with progress in attachment loss. [ABSTRACT FROM AUTHOR]