Neopterin Levels in Periodontitis and after Nonsurgical Periodontal Therapy: Evaluation of Gingival Crevicular Fluid, Oral Fluid, Serum and Urinary Samples—A Case-Control Study

Periodontitis is a chronic inflammatory disease that leads to the destruction of the tooth-supporting tissues with complex immune response. Neopterin (Np), secreted via activated macrophages, is considered a biomarker of cellular immunity. The aim of this study was to evaluate the impact of periodontitis and nonsurgical periodontal therapy. Np gingival crevicular fluid (GCF), oral fluid, serum and urine levels were compared in subjects with periodontitis before periodontal treatment, three months after and in a healthy control. Np GCF concentrations in the study group after treatment were significantly higher than the control group (p = 0.038). The GCF total amount (amount of substance) was significantly higher in the study group before periodontal treatment than in the control group (p = 0.001) and higher than the levels taken after treatment collection (p = 0.024). The oral fluid Np concentrations in the study group after treatment were significantly increased compared to the before treatment concentrations (p = 0.020). The same trend was observed in the urine samples. Significant correlation was found between the serum and oral fluid Np concentrations (p = 0.001, ρ = 0.40). Our results confirm the impact of cellular immunity and macrophages on periodontitis and on the resolution of periodontal inflammation. The presence of neopterin in oral fluid most likely originates in the serum

Koncentrace neopterinu, kynureninu a tryptofanu v sekretu z rány u pacientů s karcinomem prsu a maligním melanomem: pilotní studie

The aim of the present pilot study was to investigate the concentrations of neopterin, kynurenine and tryptophan in wound secretion in patients undergoing surgery for breast cancer or malignant melanoma. Twenty-two patients, 16 females and 6 males, undergoing surgery for breast cancer (n = 15) or malignant melanoma (n = 7) were evaluated. Neopterin, kynurenine and tryptophan were determined using a high-performance liquid chromatography method. When the concentrations in wound secretions from the primary breast tumor and the axilla were compared, the neopterin/tryptophan ratio was significantly higher in the tumor wound secretions (0.92 +/- 0.41 vs. 0.61 +/- 0.14 mmol/mol; p = 0.049), but no significant differences were observed in neopterin (49.2 +/- 28.6 vs. 31.5 +/- 11.1 nmol/L), tryptophan (52.9 +/- 13.0 vs. 51.2 +/- 13.3 mu mol/L) and kynurenine concentrations 5.97 +/- 7.49 vs. 5.34 +/- 6.25 mu mol/L) and kynurenine/tryptophan ratio (108.1 +/- 107.7 vs. 103.5 +/- 106.7 mmol/mol). No marked differences were noted in neopterin, tryptophan and kynurenine concentrations and kynurenine/tryptophan and neopterin/tryptophan ratios in sequential samples from the axilla of breast cancer patients obtained on days 1 and 2. In conclusion, present data demonstrate that the measurement of neopterin, kynurenine and tryptophan can be used to monitor local immune response after cancer surgery.Cílem prezentované pilotní studie bylo prověřit koncentrace neopterinu, kynureninu a tryptofanu v sekretu z operačních ran u pacientů léčených pro karcinom prsu nebo maligní melanom. Byl vyhodnocen vzorek od 22 pacientů (16 žen a 6 mužů), 15 léčených pro karcinom prsu, 7 pro maligní melanom. Hladina neopterinu, kynureninu a tryptofanu byla stanovena metodou high-performance tekutinové chromatografie. Byl prokázán statisticky signifikantně vyšší poměr neopterin / tryptofan v seromech z operačních ran v prsu v porovnání s ranou v axile (0,92 ± 0,41 vs. 0,61 ± 0,14 mmol / mol, p = 0,049). Koncentrace neopterinu (49,2 ± 28,6 vs. 31,5 ± 11,1 nmol / l), tryptofanu (52,9 ± 13,0 vs. 51,2 ± 13,3 μmol / L), kynureninu (5,97 ± 7,49 vs. 5,34 ± 6,25 umol / 1) a poměr kynurenin / tryptofan (108,1 ± 107,7 vs. 103,5 ± 106,7 mmol / mol) nebyla významně statisticky rozdílná. Dále nebyly zaznamenány statisticky významné rozdíly v koncentraci neopterinu, tryptofanu a kynureninu a poměru kynurenin / tryptofan a neopterin / tryptofan ve vzorcích z ran získaných v dalších pooperačních dnech. Prezentované údaje ukazují, že měření koncentrací neopterinu, kynureninu a tryptofanu může být použito k monitorování lokální imunitní odpovědi po operačním výkonu u onkologických pacienů

Mitochondrial Respiration of Platelets: Comparison of Isolation Methods

Multiple non-aggregatory functions of human platelets (PLT) are widely acknowledged, yet their functional examination is limited mainly due to a lack of standardized isolation and analytic methods. Platelet apheresis (PA) is an established clinical method for PLT isolation aiming at the treatment of bleeding diathesis in severe thrombocytopenia. On the other hand, density gradient centrifugation (DC) is an isolation method applied in research for the analysis of the mitochondrial metabolic profile of oxidative phosphorylation (OXPHOS) in PLT obtained from small samples of human blood. We studied PLT obtained from 29 healthy donors by high-resolution respirometry for comparison of PA and DC isolates. ROUTINE respiration and electron transfer capacity of living PLT isolated by PA were significantly higher than in the DC group, whereas plasma membrane permeabilization resulted in a 57% decrease of succinate oxidation in PA compared to DC. These differences were eliminated after washing the PA platelets with phosphate buffer containing 10 mmol·L−1 ethylene glycol-bis (2-aminoethyl ether)-N,N,N′,N′-tetra-acetic acid, suggesting that several components, particularly Ca2+ and fuel substrates, were carried over into the respiratory assay from the serum in PA. A simple washing step was sufficient to enable functional mitochondrial analysis in subsamples obtained from PA. The combination of the standard clinical PA isolation procedure with PLT quality control and routine mitochondrial OXPHOS diagnostics meets an acute clinical demand in biomedical research of patients suffering from thrombocytopenia and metabolic diseases