Effect of N-Acetylglucosamine on Function of Peritoneal Leukocytes

1999 ◽  
Vol 19 (2_suppl) ◽  
pp. 365-369 ◽  
Author(s):  
Malgorzata Kuzlan-Pawlaczyk ◽  
Krzysztof Pawlaczyk ◽  
Katarzyna Wieczorowska Tobis ◽  
Alicja Polubinska ◽  
Justyna Wisniewska ◽  
...  
Keyword(s):  
Peritoneal Dialysis ◽  
Cell Count ◽  
Ex Vivo ◽  
Interleukin 1 ◽  
Differential Cell Count ◽  
Dialysis Solution ◽  
Peritoneal Leukocytes ◽  
Factor Α

Objective To compare effects of N-acetylglucosamine (NAG) -based and glucose-based dialysis fluids on the function of peritoneal leukocytes in conditions of peritoneal dialysis. Design In vitro experiments on ex vivo isolated rat peritonealleukocytes. Materials Peritoneal leukocytes were isolated from rats on chronic peritoneal dialysis. On alternate days, fluid exchanges were performed with NAG-based or glucosebased dialysis solutions. After a 4-hour dwell, dialysate was drained and peritoneal leukocytes were incubated in vitro :I= lipopolysaccharide (LPS). Production of nitrites (index of NO synthesis), tumor necrosis factor α (TNFα), interleukin-1 β (IL -1 β), and interferon gamma (IFN-y) by unstimulated or stimulated peritoneal leukocytes originating from NAG-based or glucose-based fluid was measured. Results Dialysate cell count was lower during exchanges with NAG-based fluid (2113 :I= 615 cells/μL) as compared to glucose-based fluid (3643 :I= 1108 cells/μL; p < 0.01). Differential cell count was similar in both studied groups. Unstimulated peritoneal leukocytes from NAGbased dialysate produced more NO (nitrites) (0.65 ± 0.07 μmol per 106 cells) than did cells from glucose-based dialysate (0.26 :I= 0.09 μmol per 106 cells, p < 0.01). Stimulated peritoneal leukocytes from NAG-based dialysate produced more cytokines than did cells from glucose-based dialysate: TNFα, 135.2 ± 37.0 pg versus 70.2 :I= 21.8 pg per 106 cells respectively, p < 0.01; IL -1 β, 143.2 :I= 60.9 pg versus 99.1 :I= 22.4 pg per 106 cells respectively, p < 0.05; IFN-y, 16.2:1= 12.5 pg versus 6.0:1= 1.8 pg per 106 cells respectively, p <0.01. Conclusions We demonstrated that rat peritonealleukocytes exposed in vivoto NAG-based dialysis fluid have better ability to produce inflammatory mediators than do peritoneal leukocytes from the same donor, but exposed in vivo to glucose-based dialysis solution.

scholarly journals Effect of the proinflammatory cytokine TNF-α on intestinal riboflavin uptake: inhibition mediated via transcriptional mechanism(s)

2018 ◽  
Vol 315 (5) ◽  
pp. C653-C663 ◽  
Author(s):  
Kasin Yadunandam Anandam ◽  
Omar A. Alwan ◽  
Veedamali S. Subramanian ◽  
Padmanabhan Srinivasan ◽  
Rubina Kapadia ◽  
...  
Keyword(s):  
Ex Vivo ◽  
Significant Inhibition ◽  
Mrna Levels ◽  
In Vivo Models ◽  
Uptake Inhibition ◽  
Tnf Α ◽  
Vitro Model ◽  
Factor Α

Riboflavin (RF), is essential for normal cellular metabolism/function. Intestinal RF absorption occurs via a specific carrier-mediated process that involves the apical transporter RFVT-3 ( SLC52A3) and the basolateral RFVT-1 (SLC52A1). Previously, we characterized different cellular/molecular aspects of the intestinal RF uptake process, but nothing is known about the effect of proinflammatory cytokines on the uptake event. We addressed this issue using in vitro, ex vivo, and in vivo models. First, we determined the level of mRNA expression of the human (h)RFVT-3 and hRFVT-1 in intestinal tissue of patients with inflammatory bowel disease (IBD) and observed a markedly lower level compared with controls. In the in vitro model, exposing Caco-2 cells to tumor necrosis factor-α (TNF-α) led to a significant inhibition in RF uptake, an effect that was abrogated upon knocking down TNF receptor 1 (TNFR1). The inhibition in RF uptake was associated with a significant reduction in the expression of hRFVT-3 and -1 protein and mRNA levels, as well as in the activity of the SLC52A3 and SLC52A1 promoters. The latter effects appear to involve Sp1 and NF-κB sites in these promoters. Similarly, exposure of mouse small intestinal enteroids and wild-type mice to TNF-α led to a significant inhibition in physiological and molecular parameters of intestinal RF uptake. Collectively, these findings demonstrate that exposure of intestinal epithelial cells to TNF-α leads to inhibition in RF uptake and that this effect is mediated, at least in part, via transcriptional mechanism(s). These findings may explain the significantly low RF levels observed in patients with IBD.

Pre-Clinical Biocompatibility Testing of Peritoneal Dialysis Solutions

2000 ◽  
Vol 20 (5_suppl) ◽  
pp. 5-9 ◽  
Author(s):  
C.J. Holmes
Keyword(s):  
Peritoneal Dialysis ◽  
Screening Program ◽  
Ex Vivo ◽  
Biological Response ◽  
Cell Types ◽  
Hierarchical Level ◽  
Clinical Phase ◽  
Biocompatibility Testing

Pre-clinical biocompatibility testing of peritoneal dialysis (PD) solutions has become an integral part of new solution development. The construction of a pre-clinical screening program for solution biocompatibility should take a hierarchical approach, starting with in vitro cell viability and function assays. The selection of cell types and assay systems for the in vitro studies should be broad enough to permit a balanced interpretation. Whenever possible, animal models are recommended for the next hierarchical level of testing, followed by human ex vivo study designs. Designs of the latter sort provide evidence that a new solution formulation is exerting an altered biological response in vivo; the response is not purely an in vitro artifact or restricted to a given animal species. This article discusses the various approaches available for biocompatibility testing during the pre-clinical phase of solution development, with an emphasis on the advantages and drawbacks of each method.

L-2-Oxothiazolidine-4-Carboxylate: An Agent that Modulates Lipopolysaccharide-Induced Peritonitis in Rats

2002 ◽  
Vol 22 (3) ◽  
pp. 293-300 ◽  
Author(s):  
Katarzyna Korybalska ◽  
Katarzyna Wieczorowska–Tobis ◽  
Alicja Polubinska ◽  
Justyna Wisniewska ◽  
James Moberly ◽  
...  
Keyword(s):  
Peritoneal Dialysis ◽  
Peritoneal Cell ◽  
Cellular Damage ◽  
Dialysis Fluid ◽  
Acute Peritonitis ◽  
Cell Counts ◽  
Dialysis Solution ◽  
Peritoneal Leukocytes ◽  
Peritoneal Function

Objective L-2-Oxothiazolidine-4-carboxylate (OTZ), a cysteine precursor, is a substrate for intracellular glutathione synthesis. As shown previously, OTZ prevents free-radical induced cellular damage during in vitro simulation of peritoneal dialysis. In the present study, we examined the effect of adding OTZ to peritoneal dialysis solution on peritoneal function and structure during lipopolysaccharide (LPS)-induced peritonitis in rats. In addition, we studied the effects of pretreatment with OTZ (given orally) on the effects of LPS-induced peritonitis in rats. Methods Peritonitis was induced in rats by adding LPS (5 μg/mL) to the dialysis fluid. For acute experiments, rats were exposed to a single infusion of dialysis solution containing LPS or to LPS plus 5 mmol/L OTZ; peritoneal cell counts and permeability were determined after 4 hours. Alternatively, rats were pretreated with OTZ added to the drinking water (0.1%) for 10 days prior to infusion of LPS. For chronic experiments, peritoneal dialysis was performed over a 3-week period in rats with implanted peritoneal catheters. On days 8, 9, and 10 of the experiment, the rats were infused intraperitoneally with solution containing LPS (5 μg/mL), or LPS plus 5 mmol/L OTZ, to induce acute peritonitis. At the end of dialysis (10 days after the episodes of peritonitis), peritoneal function was assessed using a peritoneal equilibration test (PET), and peritoneal biopsies were taken to assess effects on peritoneal morphology. Results In the acute experiments, exposure to LPS led to increased peritoneal cell counts (+61% vs control, p < 0.05) and enhanced permeability of the peritoneum, leading to a loss in ultrafiltration (–63%, p < 0.0005). The glutathione concentration in peritoneal leukocytes also decreased during acute peritonitis (–31%, p < 0.05). During LPS-induced peritonitis, OTZ prevented the increase in dialysate cell count and the decrease in cellular glutathione content. Simultaneous administration of OTZ did not prevent the increased peritoneal permeability induced by LPS. However, in rats pretreated with OTZ, LPS-induced permeability to protein was significantly lower than in the nontreated animals (0.049 ± 0.011 vs 0.087 ± 0.034, p < 0.05). In the chronic experiments, LPS-induced peritonitis did not lead to any functional differences in peritoneal transport at the end of 3 weeks of dialysis. However, LPS-induced peritonitis led to increased thickness of the peritoneum and neovascularization within peritoneal interstitium compared to peritonitis-free animals. In contrast to the LPS-treated animals, the peritoneum of the rats exposed to LPS in the presence of OTZ was of a thickness similar to that in the control rats. Conclusion Supplementation of dialysis fluid with OTZ prevented changes in cellular glutathione levels and dialysate cell counts during acute peritonitis in rats. During chronic dialysis in rats exposed to intermittent peritonitis episodes, OTZ prevented increased thickening and neovascularization of the peritoneum. Our results suggest this may help to protect the peritoneal membrane during episodes of peritonitis.

Elevated IL-1β contributes to antibody suppression produced by stress

2002 ◽  
Vol 93 (1) ◽  
pp. 207-215 ◽  
Author(s):  
Albert Moraska ◽  
Jay Campisi ◽  
Kien T. Nguyen ◽  
Steven F. Maier ◽  
Linda R. Watkins ◽  
...  
Keyword(s):  
Innate Immunity ◽  
Acute Stress ◽  
Ex Vivo ◽  
Interleukin 1 ◽  
Keyhole Limpet Hemocyanin ◽  
Acquired Immunity ◽  
Sprague Dawley

Acute stressor exposure can facilitate innate immunity and suppress acquired immunity. The present study further characterized the potentiating effect of stress on innate immunity, interleukin-1β (IL-1β), and demonstrated that stress-induced potentiation of innate immunity may contribute to the stress-induced suppression of acquired immunity. The long-term effect of stress on IL-1β was measured by using an ex vivo approach. Sprague-Dawley rats were challenged with lipopolysaccharide (LPS) in vivo, and the IL-1β response was measured in vitro. Splenocytes, mesenteric lymphocytes, and peritoneal cavity cells had a dose- and time-dependent ex vivo IL-1β response to LPS. Rats that were exposed to inescapable shock (IS, 100 1.6 mA, 5-s tail shocks, 60-s intertrial interval) and challenged with a submaximal dose of LPS 4 days later had elevated IL-1β measured ex vivo. To test whether the acute stress-induced elevation in IL-1β contributes to the long-term suppression in acquired immunity, IL-1β receptors were blocked for 24 h after stress. Serum anti-keyhole limpet hemocyanin (KLH) immunoglobulin (Ig) was measured. In addition, the acute elevation (2 h post-IS) of splenic IL-1β in the absence of antigen was verified. Interleukin-1 receptor antagonist prevented IS-induced suppression in anti-KLH Ig. These data support the hypothesis that stress-induced increases in innate immunity (i.e., IL-1β) may contribute to stress-induced suppression in acquired immunity (i.e., anti-KLH Ig).

Inhibition of Caspases Improves Bacterial Clearance in Experimental Peritonitis

2003 ◽  
Vol 23 (2) ◽  
pp. 123-126 ◽  
Author(s):  
◽  
Marina Penélope Catalan ◽  
Jaime Esteban ◽  
Dolores Subirá ◽  
Jesús Egido ◽  
...  
Keyword(s):  
Peritoneal Dialysis ◽  
Bacterial Clearance ◽  
Experimental Peritonitis ◽  
Dialysis Solution ◽  
Peritoneal Leukocytes ◽  
Peritoneal Dialysis Solution ◽  
Dialysis Solutions ◽  
Number Of Bacteria ◽  
Peritoneal Dialysis Solutions

Background Inhibition of caspases improves the antibacterial capacity of leukocytes cultured with peritoneal dialysis solutions, and improves the prognosis of septic, polymicrobial experimental peritonitis. Objective To test whether inhibition of caspases alters the evolution of peritonitis in the presence of peritoneal dialysis solution. Design 32 mice were assigned to therapy with either the pan-caspase inhibitor zVAD or vehicle for 48 hours following infection with Staphylococcus aureus, in the presence of lactate-buffered, 4.25% glucose peritoneal dialysis solution. 16 mice received vehicle in phosphate-buffered saline. Main Outcome Measure Number of bacteria recovered from the peritoneum at 48 hours. Results Peritoneal dialysis solution accelerated leukocyte apoptosis. zVAD decreased the number of apoptotic peritoneal leukocytes and the number of bacteria recovered from the peritoneum at 48 hours (zVAD 2.8 ± 0.3 vs vehicle 3.9 ± 0.2 log colony forming units of S. aureus, p = 0.007). Conclusions Inhibition of caspases accelerates peritoneal bacterial clearance in the presence of peritoneal dialysis solutions in vivo in the experimental setting. Inhibition of caspases should be explored as a mean to accelerate recovery following peritonitis in the clinical setting.

Prevention of Peritoneal Sclerosis: A New Proposal to Substitute Glucose with Carnitine Dialysis Solution (Biocompatibility Testing in Vitro and in Rabbits)

2005 ◽  
Vol 28 (2) ◽  
pp. 177-187 ◽  
Author(s):  
E. Gaggiotti ◽  
A. Arduini ◽  
M. Bonomini ◽  
G. Valentini ◽  
G. Sacchi ◽  
...  
Keyword(s):  
Peritoneal Dialysis ◽  
Dialysis Solution ◽  
Biocompatibility Testing ◽  
Peritoneal Dialysis Solution ◽  
High Doses ◽  
Near Future ◽  
Dialysis Solutions ◽  
Peritoneal Dialysis Solutions

Aim Commercial glucose peritoneal dialysis solutions expose the peritoneum to hyperosmolar glucose containing variable amounts of non-enzymic breakdown products of glucose. These solutions are toxic for the peritoneum. The aim of the present study is to compare in vitro and in vivo characteristics of a new dialysis solution containing carnitine, a naturally occurring compound, as substitute of glucose. Material and Methods We compared in vitro and in the rabbit a new peritoneal dialysis solution containing carnitine, with two standard bicarbonate glucose peritoneal dialysis solutions and a solution containing icodextrin. Results In vitro and in vivo the solution containing carnitine seems to be more biocompatible than standard glucose solutions and those containing icodextrin. Conclusions In our study the peritoneal dialysis solution containing carnitine seems to prevent the mesothelial changes observed with solutions containing glucose. Since carnitine has been extensively studied and seems to be well tolerated by hemodialysis patients, even at high doses for long periods, clinical trials in humans may be planned in the near future.

Ca2+ Concentration in the Peritoneal Dialysis Solution Regulates Peritoneal Fibroblast Proliferation and Peritoneal Macrophage and Lymphocyte Cytokine Release

1993 ◽  
Vol 13 (2_suppl) ◽  
pp. 41-43 ◽  
Author(s):  
Silvia Carozzi ◽  
Maria Grazia Nasini
Keyword(s):  
Peritoneal Dialysis ◽  
Interleukin 1 ◽  
Peritoneal Macrophages ◽  
Peritoneal Fibrosis ◽  
Ifn Gamma ◽  
Dialysis Solution ◽  
Peritoneal Dialysis Solution ◽  
Uremic Patients ◽  
And Control

Peritoneal fibrosis remains one of the major causes of dropout In continuous ambulatory peritoneal dialysis (CAPD), because it reduces ultrafiltration capacity. Since studies In vitro have demonstrated that cytoplasmic Ca2+ regulates the proliferation of most cell lines and the release of cytokines from immune cells, we evaluated In 8 uremic patients at the start of CAPD and in 4 control patients the effects in vitro of different peritoneal dialysis solution Ca concentrations (1, 1.25, 1.75, and 2 mmol/L) on peritoneal fibroblast (PF) proliferation, peritoneal macrophages (PMΦ), and peritoneal lymphocyte (Ply) release of interleukin-1 (11–1) and Interferon-gamma (IFN-gamma) (cytokines which are known to induce PF proliferation), and cytoplasmic Ca2+ concentration in PF, PMΦ, and Ply. Results showed that in both the uremic and control patients, increasing the dose of Ca2+ In the medium induced a dose-dependent increase in PF proliferation and the release of IL-1 and IFN-gamma from PMΦ and Ply. Meanwhile, the cytoplasmic parameters PF, PMΦ, and Ply Ca2+ in the uremic patients were below normal; they exceeded the norm with a Ca2+ concentration of 1.75 and 2 mmol/L and were normal with a Ca2+ concentration of 1.25 mmol/L. These data suggest that In CAPD patients the use of a physiological Ca peritoneal dialysis solution (1 and 1.25 mmol/L) may be useful in reducing the proliferation of PF and the production of IL-1 and IFN-gamma thus preventing peritoneal sclerosis.

scholarly journals Chondroprotective Properties of Human-Enriched Serum Following Polyphenol Extract Absorption: Results from an Exploratory Clinical Trial

Nutrients ◽  
2019 ◽  
Vol 11 (12) ◽  
pp. 3071
Author(s):  
Fabien Wauquier ◽  
Elsa Mevel ◽  
Stephanie Krisa ◽  
Tristan Richard ◽  
Josep Valls ◽  
...  
Keyword(s):  
Articular Chondrocytes ◽  
Ex Vivo ◽  
Interleukin 1 ◽  
Clinical Approach ◽  
Age Related ◽  
Nutritional Strategy ◽  
Sparing Effect ◽  
The Relationship

Polyphenols are widely acknowledged for their health benefits, especially for the prevention of inflammatory and age-related diseases. We previously demonstrated that hydroxytyrosol (HT) and procyanidins (PCy), alone or in combination, drive preventive anti-osteoathritic effects in vivo. However, the lack of sufficient clinical evidences on the relationship between dietary phytochemicals and osteoarthritis remains. In this light, we investigated in humans the potential osteoarticular benefit of a grapeseed and olive extract (OPCO) characterized for its hydroxytyrosol (HT) and procyanidins (PCy) content. We first validated, in vitro, the anti-inflammatory and chondroprotective properties of the extract on primary cultured human articular chondrocytes stimulated by interleukin-1 beta (IL-1 β). The sparing effect involved a molecular mechanism dependent on the nuclear transcription factor-kappa B (NF-κB) pathway. To confirm the clinical relevance of such a nutritional strategy, we designed an innovative clinical approach taking into account the metabolites that are formed during the digestion process and that appear in circulation after the ingestion of the OPCO extract. Blood samples from volunteers were collected following ingestion, absorption, and metabolization of the extract and then were processed and applied on human primary chondrocyte cultures. This original ex vivo methodology confirmed at a clinical level the chondroprotective properties previously observed in vitro and in vivo.

scholarly journals Cell population, viability, and some key immunomodulatory molecules in different milk somatic cell samples in dairy cows

2009 ◽  
Vol 76 (3) ◽  
pp. 356-364 ◽  
Author(s):  
Amandine Baumert ◽  
Rupert M. Bruckmaier ◽  
Olga Wellnitz
Keyword(s):  
Mammary Gland ◽  
Somatic Cell ◽  
Cell Count ◽  
Mrna Expression ◽  
Immune Cells ◽  
Ex Vivo ◽  
Death Receptor ◽  
Cell Loss

Immune cells in the milk are most important in combating pathogens that invade the mammary gland. This study investigated the immune competence and viability of somatic milk cells that are already resident in milk and udders free of infection. Cells were studied in freshly removed milk to simulate conditions in the udder. Effects of incubation, cell preparation, and immunological stimulation with 0·5 μg/ml lipopolysaccharide (LPS) fromEscherichia coliwere analysed. Viability and differential counts of milk cells between high and low somatic cell count (SCC) quarters, and cisternal and alveolar milk with and without LPS stimulation were compared. Incubation and preparation of cells caused a cell loss which further increased with time independently of SCC and milk fraction. The viability of these cells was stable until 3 h post incubation and decreased until 6 h. Cell populations differed between both investigations, but did not change during the course of the experiment. mRNA expression of immune and apoptosis factors of the cells, measured by qPCR, did not change substantially: mRNA expression of caspase 3, Toll like receptor 4, and GM-CSF did not change, whereas the expression of the death receptor Fas/APO-1 (CD95), lactoferrin and lysozyme was decreased at 6 h. Cyclooxygenase-2 and TNF-α mRNA expression were decreased after 6 h of LPS treatment. In comparison with other studies in vivo or in vitro (in cell culture), in this study where cells are studied ex vivo (removed from the udder but kept in their natural environment, the milk) resident milk cells seem to be more vulnerable, less viable, less able to respond to stimulation, and thus less immune competent compared with cells that have freshly migrated from blood into milk after pathogen stimulation. The cell viability and differential cell count differed between high- and low-SCC milk and between cisternal and alveolar milk depending on the individual cow. In conclusion, the results support the view that for a most effective defence against invading pathogens the mammary gland is reliant on the recruitment of fresh immune cells from the blood.

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