The CAMKK/AMPK Pathway Contributes to Besnoitia besnoiti-Induced NETosis in Bovine Polymorphonuclear Neutrophils.

Besnoitia besnoiti is an obligate intracellular apicomplexan parasite and the causal agent of bovine besnoitiosis. Bovine besnoitiosis has a considerable economic impact in Africa and Asia due to reduced milk production, abortions, and bull infertility. In Europe, bovine besnoitiosis is classified as an emerging disease. Polymorphonuclear neutrophils (PMN) are one of the most abundant leukocytes in cattle blood and amongst the first immunological responders toward invading pathogens. In the case of B. besnoiti, bovine PMN produce reactive oxygen species (ROS), release neutrophil extracellular traps (NETs), and show increased autophagic activities upon exposure to tachyzoite stages. In that context, the general processes of NETosis and autophagy were previously reported as associated with AMP-activated protein kinase (AMPK) activation. Here, we study the role of AMPK in B. besnoiti tachyzoite-induced NET formation, thereby expanding the analysis to both upstream proteins, such as the calcium/calmodulin-dependent protein kinase kinase 2 (CAMKK), and downstream signaling and effector molecules, such as the autophagy-related proteins ULK-1 and Beclin-1. Current data revealed early AMPK activation (<30 min) in both B. besnoiti-exposed and AMPK activator (AICAR)-treated bovine PMN. This finding correlated with upstream responses on the level of CAMKK activation. Moreover, these reactions were accompanied by an augmented autophagic activity, as represented by enhanced expression of ULK-1 but not of Beclin-1. Referring to neutrophil effector functions, AICAR treatments induced both AMPK phosphorylation and NET formation, without affecting cell viability. In B. besnoiti tachyzoite-exposed PMN, AICAR treatments failed to affect oxidative responses, but led to enhanced NET formation, thereby indicating that AMPK and autophagic activation synergize with B. besnoiti-driven NETosis.

Mechanical Properties of DPPC-POPE Mixed Langmuir Monolayers.

The mechanical properties of lipid monolayers and their responses to shear and compression stresses play an important role in processes such as breathing and eye blinking. We studied the mechanical properties of Langmuir monolayers of a model mixture, composed of an unsaturated lipid, 1-palmitoyl-2-oleoyl-sn-glycero-phosphoethanolamine (POPE), and a saturated lipid, 1,2-dipalmitoyl-sn-glycero-phosphocholine (DPPC). We performed isothermal compressions and sinusoidal shear deformations of these mixed monolayers. Also, the different phases were observed with Brewster angle microscopy. We found that the mechanical behavior is affected by the miscibility of both lipids. In the two-phase region, the compression elastic modulus increases with the amount of the LC phase but does not follow the predictions of a simple effective medium model. The discrepancies arise from the fact that, upon compression, the domains grow at a rate faster than the compression rate but not fast enough to reach thermodynamic equilibrium. Before reaching the LC phase, domain percolation is observed and compression and shear moduli become equal to those of the pure LC phase. Most of the monolayers behave as viscoelastic fluids at the frequencies investigated. A minimum in the compression modulus and shear viscosity was observed for mixtures close to equimolar composition, with the minimum being accompanied by a change in domain shapes.

Proposal of a New Specific Cardiopulmonary Exercise Test for Taekwondo Athletes.

The objective of this study is to evaluate the cardiorespiratory variables of Taekwondo athletes while performing incremental exercise test on an ergometer using a ramp protocol and to propose a specific protocol for assessing these physiological variables during Taekwondo practice. Fourteen athletes participated in 2 incremental exercise tests: a treadmill exercise test (TREADtest) and a Taekwondo-specific exercise test (TKDtest). The TKDtest consists in 1-minute stages of kicks with an incremental load between then. The subjects perform kicks each time a sound signal was heard. Heart rate (HR), oxygen uptake (V[Combining Dot Above]O2), and their reserve correspondents (V[Combining Dot Above]O2R and reserve heart rate [HRR]) were divided into quartiles to verify their kinetics along the tests. Significant difference between 2 tests was found only for V[Combining Dot Above]O2R (p = 0.03). Regarding the quartiles, significant differences were found for HR in the first (p = 0.030) and second (p = 0.003). Analyzing the regression curves, significant differences were found for HR for intercept (p = 0.01) and slope (p = 0.05) and HRR for slope (p = 0.02). Analysis showed that significant reliability, with intraclass correlation coefficient (ICC), was found for the V[Combining Dot Above]O2peak (ICC = 0.855, p = 0.003), V[Combining Dot Above]O2 in ventilatory thresholds 1 (ICC = 0.709, p = 0.03) and 2 (ICC = 0.848, p = 0.003). Bland-Altman analyses reported a mean difference ± the 95% limits of agreement of 2.2 ± 8.4 ml·kg·min to V[Combining Dot Above]O2peak. The TKDtest is reliable for measurement of cardiorespiratory variables, and the behavior of these variables differs mainly from TREADtest, probably because of the motor task performed.

Composition of the HLA-DR-associated human thymus peptidome.

Major histocompatibility complex class II (MHC-II) molecules bind to and display antigenic peptides on the surface of antigen-presenting cells (APCs). In the absence of infection, MHC-II molecules on APCs present self-peptides and interact with CD4(+) T cells to maintain tolerance and homeostasis. In the thymus, self-peptides bind to MHC-II molecules expressed by defined populations of APCs specialised for the different steps of T-cell selection. Cortical epithelial cells present peptides for positive selection, whereas medullary epithelial cells and dendritic cells are responsible for peptide presentation for negative selection. However, few data are available on the peptides presented by MHC molecules in the thymus. Here, we apply mass spectrometry to analyse and identify MHC-II-associated peptides from five fresh human thymus samples. The data show a diverse self-peptide repertoire, mostly consisting of predicted MHC-II high binders. Despite technical limitations preventing single cell population analyses of peptides, these data constitute the first direct assessment of the HLA-II-bound peptidome and provide insight into how this peptidome is generated and how it drives T-cell repertoire formation.

Shear rheology of lipid monolayers and insights on membrane fluidity.

The concept of membrane fluidity usually refers to a high molecular mobility inside the lipid bilayer which enables lateral diffusion of embedded proteins. Fluids have the ability to flow under an applied shear stress whereas solids resist shear deformations. Biological membranes require both properties for their function: high lateral fluidity and structural rigidity. Consequently, an adequate account must include, in addition to viscosity, the possibility for a nonzero shear modulus. This knowledge is still lacking as measurements of membrane shear properties have remained incomplete so far. In the present contribution we report a surface shear rheology study of different lipid monolayers that model distinct biologically relevant situations. The results evidence a large variety of mechanical behavior under lateral shear flow.

Peak oxygen uptake after the 80s as a survival predictor.

PURPOSE: Peak oxygen uptake (VO2peak) is a crucial health marker, extensively studied in adults for its prognostic value. However, its significance in the older persons, especially octogenarians, remains underexplored due to limited representation in research. This study aims to assess the predictive power of VO2peak for survival in individuals aged 80 and above. METHODS: We included individuals aged 80 or older who underwent cardiopulmonary exercise tests at a single center. Mortality rates were compared based on VO2peak relative to 80% of predicted values (%VO2peak). We employed three multivariate Cox regression models: Model 1 (unadjusted), Model 2 (adjusted for age) and Model 3 (adjusted for age and stroke). RESULTS: Among 188 participants (mean age 83.3 ± 3 years, 68.9% male), 22 (11.7%) passed away during a median follow-up of 494 days. Non-survivors tended to be older with lower VO2peak and %VO2peak. All models demonstrated associations between %VO2peak ≤ 80% and mortality: HR = 3.19 (95% CI: 1.30-7.86, p = 0.011) for M1; HR = 3.12 (95% CI: 1.26-7.74, p = 0.013) for M2 and HR = 2.80 (95% CI: 1.11-7.06, p = 0.028) for M3. CONCLUSION: In the context of an aging population, this study underscores the enduring significance of VO2peak as a survival predictor among the older person, including octogenarians. These findings carry profound implications for tailoring healthcare strategies to address the evolving demographic landscape.

Shear and compression rheology of Langmuir monolayers of natural ceramides: solid character and plasticity.

The present work addresses the fundamental question of membrane elasticity of ceramide layers with a special focus on the plastic regime. The compression and shear viscoelasticity of egg-ceramide Langmuir monolayers were investigated using oscillatory surface rheology in the linear regime and beyond. High compression and shear moduli were measured at room temperature-a clear signature for a solid behavior. At deformations larger than one per mill, egg-ceramide monolayers display plastic features characterized by a decrease of the storage modulus followed by a viscous regime typical of fluid lipids. This behavior is accompanied by a marked decrease of the loss modulus with increasing stress above a yield point. The results permit to univocally classify ceramide monolayers as 2D solids able to undergo plastic deformations, at the difference of typical fluid lipid monolayers. These unusual features are likely to have consequences in the mechanical behavior of ceramide-rich emplacements in biological membranes.

Physiological effects of exercising at different intensities wearing surgical or double-layer cotton facemasks compared to not wearing a mask.

Since the beginning of the SARS-CoV-2 pandemic, the community use of facemasks has been widely recommended. However, their use during exercise has raised safety concerns. Thus, we compared the physiological differences between exercising wearing a surgical (SM) or a double-layer-cotton (DLC) facemask and not wearing a mask (NM). Sixteen volunteers underwent 4 bouts of cycling-based exercise, which consisted of two different intensities: light-to-moderate and moderate-to-high. Facemasks were used as follows: bout-1 and 4: NM; bout-2: SM or DLC and bout-3: DLC or SM. Ventilatory, metabolic, pulmonary gas exchange (PGE) and perceptual variables were collected. At both exercise intensities compared to NM, both facemasks induced similar ventilatory adaptations, increasing inspiratory time and tidal volume and decreasing breathing frequency. Effect sizes (ES) were larger for DLC than for SM. At moderate-to-high, both facemasks reduced the minute ventilation, whereas at light-to-moderate, it was only seen with DLC. End tidal and mixed CO2 pressures, as well as the difference between them, increased with both facemasks. Again, ES was larger for DLC than SM. No relevant oxygen saturation drop was observed with both facemaks and exercise intensities. A small ES increament in VO2 and VCO2 were seen with both facemasks. Effort perception increased at moderate-to-high for both exercise intensities, buth larger EF were with DLC than SM . DLC increased facial temperature during both exercise intensities. In conclusion, ventilatory adjustments imposed during facemask exercise influenced PGE and metabolic and perceptual changes. Larger ES were mostly seen for DLC than SM.Abbreviations: Bf: Breathing frequency.; CPET: Cardiopulmonary exercise test.; CI: Confidence interval.; DLC: Double-layer cotton.; ETCO2: End tidal CO2 pressure.; ES: Effect size.; ΔET-PECO2: Difference between ETCO2 and PECO2.; FMMT: Facemask microclimate temperature.; HR: Heart rate.; IQR: Interquartile range.; NM: No mask.; PECO2: Mixed-expired CO2 pressure.; RER: Respiratory exchange ratio.; RPE: Rate of perceived effort.; SD: Standard deviation.; SM: Surgical Mask.; SpO2: Oxygen saturation.; STP: Subjective thermal perception.; Ti/TTOT: Duty cycle.; VE: Minute ventilation.; VCO2: Carbon dioxide output.; VO2: Oxygen uptake.; VT: Tidal volume.; VT: Ventilatory threshold.HighlightsFacemasks affect the breathing pattern by changing the frequency and amplitude of pulmonary ventilation.The augmented ventilatory work increases VO2, VCO2, and RPE and promotes nonconcerning drops in SpO2 and CO2 retention.Increased inspiratory and expiratory pressure can account for the reduction in pulmonary physiological dead space.

Besnoitia besnoiti-induced neutrophil clustering and neutrophil extracellular trap formation depend on P2X1 purinergic receptor signaling.

Bovine besnoitiosis is a re-emerging cattle disease caused by the cyst-forming apicomplexan parasite Besnoitia besnoiti. Neutrophil extracellular trap (NET) formation represents an efficient innate immune mechanism of polymorphonuclear neutrophils (PMN) against apicomplexan parasites, including B. besnoiti. PMN purinergic signaling was proposed as a critical factor for NET formation. One important purinergic ligand is ATP, which is recognized as a danger signal and released into the extracellular space acting as an autocrine/paracrine signaling molecule. ATP-driven effects on PMN via the nucleotide P2 receptor family include chemotaxis, reactive oxygen species (ROS) production, and NET formation. So far, data on both PMN ATP concentrations and the role of ATP as a key modulator of purinergic signaling in B. besnoiti tachyzoite-triggered bovine NETosis is scarce. Current data showed that B. besnoiti tachyzoite exposure to bovine PMN neither changed total PMN ATP nor extracellular ATP quantities even though it significantly triggered NET formation. Moreover, B. besnoiti tachyzoite-exposed PMN revealed enhanced oxygen consumption rates (OCR) as quantified by the Seahorse metabolic analyzer. Exogenous supplementation of ATP or non-hydrolizable ATP (ATPγS) led to increased extracellular acidification rates (ECAR) but failed to alter tachyzoite-induced oxidative responses (OCR) in exposed PMN. In addition, exogenous supplementation of ATPγS, but not of ATP, boosted B. besnoiti tachyzoite-induced anchored NET formation. Referring to purinergic signaling, B. besnoiti tachyzoite-triggered anchored NET formation revealed P2X1 purinergic as receptor-dependent since it was blocked by the P2X1 inhibitor NF449 at an IC50 of 1.27 µM. In contrast, antagonists of P2Y2, P2Y6, P2X4, and P2X7 purinergic receptors all failed to affect parasite-driven NETosis. As an interesting finding, we additionally observed that B. besnoiti tachyzoite exposure induced PMN clustering in a P2X1-dependent manner. Thus, we identified P2X1 purinergic receptor as a pivotal molecule for both B. besnoiti tachyzoite-induced PMN clustering and anchored NET formation.

Magnesium production by molten salt electrolysis with liquid tin cathode and multiple effect distillation.

Low-cost clean primary production of magnesium metal is important for its use in many applications, from light-weight structural components to energy technologies. This work describes new experiments and cost and emissions analysis for a magnesium metal production process. The process combines molten salt electrolysis of MgO using MgF2-CaF2 electrolyte and a reactive liquid tin cathode, with gravity-driven multiple effect thermal system (G-METS) distillation to separate out the magnesium product, and re-use of the tin. Electrolysis experiments with carbon anodes showed current yield above 90%, while a yttria-stabilized zirconia solid oxide membrane (SOM) anode experiment showed 84% current yield. G-METS distillation is an important component of the envisioned process. It can potentially lower costs and energy use considerably compared with conventional magnesium distillation. Techno-economic analysis including detailed mass and energy balances shows that this electrolyte composition could lower costs by utilizing CaO, which is the primary impurity in MgO, as the Hall-Héroult process uses the sodium impurity in alumina. Analysis options include: raw material types (magnesite rock vs. brine or seawater), drying and calcining using electricity vs. natural gas, and carbon vs. SOM anode type. Using SOM inert anodes results in a cost premium around 10%-15%, mostly due to higher electrical energy usage resulting from membrane resistance, and reduces GHG emissions by approximately 1 kg CO2/kg Mg product. Capital and operating cost estimates, and cradle to gate greenhouse gas (GHG) emissions analysis under several raw material and process technology scenarios, show comparable costs and emissions to those of aluminum production.

Abnormal exercise adaptation after varying severities of COVID-19: A controlled cross-sectional analysis of 392 survivors.

The multisystem impairment promoted by COVID-19 may be associated with a reduction in exercise capacity. Cardiopulmonary abnormalities can change across the acute disease severity spectrum. We aimed to verify exercise physiology differences between COVID-19 survivors and SARS-CoV-2-naïve controls and how illness severity influences exercise limitation. A single-centre cross-sectional analysis of prospectively collected data from COVID-19 survivors who underwent cardiopulmonary exercise testing (CPET) in their recovery phase (x = 50[36;72] days). Patients with COVID-19 were stratified according to severity as mild [M-Cov (outpatient)] vs severe/critical [SC-Cov(inpatients)] and were compared with SARS-CoV-2-naïve controls (N-Cov). Collected information included demographics, anthropometrics, previous physical exercise, comorbidities, lung function test and CPET parameters. A multivariate logistic regression analysis was performed to identify low aerobic capacity (LAC) predictors post COVID-19. Of the 702 included patients, 310 (44.2%), 305 (43.4%) and 87 (12.4%) were N-Cov, M-Cov and SC-Cov, respectively. LAC was identified in 115 (37.1%), 102 (33.4%), and 66 (75.9%) of N-CoV, M-CoV and SC-CoV, respectively (p < 0.001). SC-Cov were older, heavier with higher body fat, more sedentary lifestyle, more hypertension and diabetes, lower forced vital capacity, higher prevalence of early anaerobiosis, ventilatory inefficiency and exercise-induced hypoxia than N-Cov. M-Cov had lower weight, fat mass, and coronary disease prevalence and did not demonstrate more CEPT abnormalities than N-Cov. After adjustment for covariates, SC-Cov was an independent predictor of LAC (OR = 2.7; 95% CI, 1.3-5.6). Almost two months after disease onset, SC-CoV presented several exercise abnormalities of oxygen uptake, ventilatory adaptation and gas exchange, including a high prevalence of LAC.Highlights Weeks after the acute disease phase, one-third of mild and three-quarters of severe and critical patients with COVID-19 presented a reduced aerobic capacity. Previous studies including SARS-CoV-1 survivors observed much lower values.A severe or critical COVID-19 case was an independent predictor for low aerobic capacity.In our sample, pre-COVID-19 exercise significantly reduced the odds of post-COVID-19 low aerobic capacity. Even severe or critical patients who exercised regularly had a prevalence of low aerobic capacity 2.5 times lower than those who did not have this routine before sickening.

Crystal-arrested phase separation.

We have studied the interplay between phase separation and crystallization in a colloid-polymer mixture along one kinetic pathway in samples which exhibit three-phase equilibrium coexistence. In analogy with atomic systems, the range of the effective attractive interaction between colloids is sufficiently long to allow for a stable liquid phase. By direct imaging in microgravity on the International Space Station, we observe a unique structure, a "crystal gel," that occurs when gas-liquid phase separation arrests due to crystallites within the liquid domain spanning the cell. From the initial onset of spinodal decomposition until arrest caused by this structure, the kinetics of phase separation remain largely unaffected by the formation of the third phase. This dynamic arrest appears to result from the stiffness of the crystalline strands exceeding the liquid-gas interfacial tension.

Evaluation of peripheral blood polymorphonuclear cell functions after an oral carbohydrate overload in obese and insulin dysregulated horses.

Obesity and insulin dysregulation (ID) are increasingly prevalent conditions in equid populations worldwide. Immune impairment is well described in humans with metabolic dysfunction and is reported but still incompletely understood in horses. This study evaluated the effect of acute induced transient hyperglycemia on apoptosis, phagocytosis and oxidative burst activity of peripheral blood polymorphonuclear cells (PMN) of lean and obese adult horses with or without insulin dysregulation. Seventeen adult horses were allocated into three groups based on their body condition score (BCS) and metabolic status: lean-insulin sensitive (lean-IS), obese-insulin sensitive (obese-IS) and obese-insulin dysregulated (obese-ID). ID was determined by insulin tolerance testing (ITT). Blood glucose elevation was induced through an infeed-oral glucose test (in-feed OGT), and all assessments of PMN functions (apoptosis, phagocytosis and oxidative burst) were done in vitro after isolation from peripheral blood before and 120 min after carbohydrate overload. Results were analyzed using a repeated measures linear mixed model with significance defined at P < 0.05. No differences in apoptosis were observed between experimental groups at any time point. Phagocytic capacity was significantly lower at baseline in the obese-ID group but increased in response to glucose administration when compared to the other two groups. Basal reactive oxygen species production in the obese-IS group differed significantly from the lean-IS and obese-ID groups and decreased significantly in response to glucose administration. Results from this study showed that both metabolic status itself, and oral glucose administration, seem to be factors that alter PMN functionality in horses, specifically phagocytosis and oxidative burst.

Solid character of membrane ceramides: a surface rheology study of their mixtures with sphingomyelin.

The compression and shear viscoelasticities of egg-ceramide and its mixtures with sphingomyelin were investigated using oscillatory surface rheology performed on Langmuir monolayers. We found high values for the compression and shear moduli for ceramide, compatible with a solid-state membrane, and extremely high surface viscosities when compared to typical fluid lipids. A fluidlike rheological behavior was found for sphingomyelin. Lateral mobilities, measured from particle tracking experiments, were correlated with the monolayer viscosities through the usual hydrodynamic relationships. In conclusion, ceramide increases the solid character of sphingomyelin-based membranes and decreases their fluidity, thus drastically decreasing the lateral mobilities of embedded objects. This mechanical behavior may involve important physiological consequences in biological membranes containing ceramides.

Piscirickettsia salmonis-Triggered Extracellular Traps Formation as an Innate Immune Response of Atlantic Salmon-Derived Polymorphonuclear Neutrophils.

Extracellular traps (ETs) are webs of DNA, citrullinated histones, anti-microbial peptides, and proteins that were not previously reported in Atlantic salmon (Salmo salar). ETs are mainly released from polymorphonuclear neutrophils (PMN) and are considered a novel PMN-derived effector mechanism against different invasive pathogens. Here, we showed that Atlantic salmon-derived PMN released ETs-like structures in vitro in response to highly pathogenic facultative intracellular rickettsial bacteria Piscirickettsia salmonis. PMN were isolated from pre-smolt Atlantic salmon and stimulated in vitro with oleic acid and P. salmonis. Extracellular DNA was measured using the PicoGreen™ dye, while immunofluorescence image analysis was used to confirm the classical components of salmonid-extruded ETs. Future studies are required to better understand the role of Atlantic salmon-derived ETs orchestrating innate/adaptive immunity and the knowledge on regulation pathways involved in this cell death process. Thus, comprehension of salmonid-derived ETs against P. salmonis might represent novel alternative strategies to improve host innate defense mechanisms of farmed salmon against closely related rickettsial bacteria, as a complement to disease prevention and control strategies.

Assessment of peripheral blood neutrophil respiratory burst, phagocytosis and apoptosis in obese non-insulin dysregulated horses.

Obesity is a highly prevalent condition in horses. Dysfunctional neutrophil activity has been reported in metabolically healthy obese humans, but minimal data exist regarding horses. The present study evaluated the effect of obesity on apoptosis, phagocytosis and oxidative burst activity of peripheral blood neutrophils from lean and obese non-insulin dysregulated horses. Seven lean (BCS, body condition score 4-6/9) and five obese (BCS 8-9) horses were enrolled in the study. All animals underwent two metabolic tests (OGT, oral glucose test; IRT, insulin response test) before their selection to ensure their metabolic status (non-insulin dysregulated). A single blood sample was obtained from each horse, and a discontinuous density gradient was carried out to isolate neutrophils. Phagocytosis, apoptosis and reactive oxygen species (ROS) production assays were performed for each animal. All statistical analyses were performed with unpaired two-tailed t-tests. Results indicate that neutrophils from obese non-insulin dysregulated horses have a significantly increased ROS production (P < .0001), with no changes observed on phagocytosis (P > .05) or apoptosis (P > .05) when compared to the control group. In conclusion, our results demonstrate that obesity per se, in absence of other endocrine disorders, alters neutrophil reactive oxygen species production. More research is needed to understand the role of obesity on the equine immune system of horses, and its role in the development of endocrine disorders.

Pre-conditioning of Equine Bone Marrow-Derived Mesenchymal Stromal Cells Increases Their Immunomodulatory Capacity.

Mesenchymal stem/stromal cells (MSCs) are increasingly explored for the treatment of degenerative and inflammatory diseases in human and veterinary medicine. One of the key characteristics of MSCs is that they modulate inflammation mainly through the secretion of soluble mediators. However, despite widespread clinical use, knowledge regarding the effector mechanisms of equine MSCs, and consequently their effectiveness in the treatment of diseases, is still unknown. The objectives of this study were to determine the mechanisms underlying inhibition of lymphocyte proliferation by equine bone marrow-derived MSCs, and to evaluate the effect of pre-conditioning of equine MSCs with different pro-inflammatory cytokines on inhibition of lymphocyte proliferation. We determined that inhibition of lymphocyte proliferation by equine MSCs depends on activity of prostaglandin-endoperoxide synthase 2 and indoleamine 2,3-dioxygenase. Additionally, pre-conditioning of MSCs with TNF-α, IFN-γ or their combination significantly increased the expression of prostaglandin-endoperoxide synthase 2, indoleamine 2,3-dioxygenase, iNOS and IL-6. This upregulation correlated with an increased inhibitory effect of MSCs on lymphocyte proliferation. In conclusion, pre-conditioning of bone marrow-derived MSC increases their inhibitory effect on lymphocyte proliferation in horses.

Equine bone marrow-derived mesenchymal stromal cells inhibit reactive oxygen species production by neutrophils.

BACKGROUND: Polymorphonuclear neutrophils (PMN) are the largest leukocyte population in the blood of most mammals including horses, and play an important defensive role in many infectious diseases. However, the mechanisms that increase PMN as one of the main cellular subsets in the defense against pathogens could also be involved in the pathophysiology of dysregulated inflammatory conditions. Mesenchymal stem/stromal cells (MSCs) are a heterogeneous population with a modulatory potential on the inflammatory response and are known to interact with nearly all cells of the immune system, including PMN. In this study, the in vitro modulation of equine bone marrow-derived MSCs on equine PMN phagocytosis, ROS production, and NETs generation was assessed. RESULTS: In co-culture with MSCs, unstimulated PMN produce less ROS (2.88 % ±â€¯1.43) than PMN in single culture (5.89 % ±â€¯2.63) (p = 0.016). Moreover, PMN co-cultured with MSCs remain conditioned to produce fewer ROS after PMA stimulation in comparison to PMN in single culture (p < 0.05). Additionally, it was found that incubation with MSC supernatant strongly inhibited ROS production (83 % ±â€¯6.35 less than control) without affecting phagocytosis or capacity for NETosis (p < 0.01). CONCLUSIONS: These results suggest a modulatory effect of equine BM-derived MSCs on PMN respiratory burst, without impairing other important microbicidal functions. This supports the potential use of equine MSCs in excessive or persistent inflammatory conditions in which neutrophils are the main effector cells.

Viscoelastic properties of doxorubicin-treated HT-29 cancer cells by atomic force microscopy: the fractional Zener model as an optimal viscoelastic model for cells.

The malignancy of cancer cells and their response to drug treatments have been traditionally studied using solely their elastic properties. However, the study of the combined viscous and elastic properties provides a richer description of the mechanics of the cell, and achieves a more precise assessment of the effect exerted by anti-cancer treatments. We used an atomic force microscope to obtain the morphological, elastic and viscous properties of HT-29 colorectal cancer cells. Changes in these parameters were observed during exposure of the cells to doxorubicin at different times. The elastic properties were analyzed using the Hertz and Sneddon models. Furthermore, we analyzed the data to study the viscoelasticity of the cells comparing the models known as the standard linear solid, fractional Zener, generalized Maxwell, and power law. A discussion about the optimal model based in the accuracy and physical assumptions for this particular system is included. From the morphological data and viscoelasticity of HT-29 cells exposed to doxorubicin, we found that some parameters were affected differently at shorter or longer exposure times. For instance, the relaxation time suggests a measure of the cell to self-heal and it was observed to increase at shorter exposure times and then to reduce for longer exposure times to the drug. The fractional Zener model better described the mechanical properties of the cell due to the reduced number of parameters and the quality of the fit to experimental data.

Interfacial shear rheology of mixed polyelectrolyte-surfactant layers.

We have studied the shear rheology of mixed surface layers containing polyelectrolytes and surfactants of opposite charges. The layers containing rigid polyelectrolytes are solidlike and can even exhibit brittle behavior. More flexible polyelectrolytes lead to more viscoelastic layers and very flexible ones to purely viscous layers. A relaxation time on the order of tens of seconds was found with the less flexible polymers. In the DNA case, it was possible to show that this relaxation time decreases with the inverse shear rate, as in three-dimensional soft solids. These short times suggest that the polymer chains might not be entangled between the surface layers containing neutral polymers.