Whole blood concentrations of fingolimod and its pharmacologically active metabolite fingolimod phosphate obtained during routine health care of patients with multiple sclerosis.

Background: Fingolimod is a first-in-class, orally administered drug indicated for the treatment of relapsing-remitting multiple sclerosis. It acts as an immunomodulator, is classified as a "disease-modifying therapy", and its main mechanism of action is the modulation of sphingosine-1-phosphate receptors. In this prospective pilot study, whole blood concentrations of fingolimod and fingolimod phosphate obtained during routine health care were measured.

Liquid chromatography-tandem mass spectrometry for determination of fingolimod and its active metabolite fingolimod phosphate in whole blood of patients with multiple sclerosis.

Fingolimod is an oral drug for the escalation of treatment of relapsing-remitting multiple sclerosis in patients with persistent disease activity on first-line drugs or in patients with rapidly progressive severe relapsing-remitting multiple sclerosis. An ultra-high-performance liquid chromatography-tandem mass spectrometry method for determining the concentrations of fingolimod and its active metabolite fingolimod phosphate in whole blood has been developed and validated. The advantages of this method are the easy, fast and cheap sample preparation using protein precipitation from blood with a mixture of acetonitrile-methanol (40:60, v/v).

Analysis of Concentrations of Monomethyl Fumarate in Patients with Multiple Sclerosis: Result from Routine Health Care.

Background: Dimethyl fumarate is used to treat patients with relapsing-remitting multiple sclerosis. After ingestion, it is rapidly hydrolyzed to the active primary metabolite monomethyl fumarate.

Objective: The main objective of our study was to analyze serum concentrations of monomethyl fumarate during routine health care in patients with multiple sclerosis treated with a fixed dose of dimethyl fumarate.

Serum teriflunomide concentrations in routine multiple sclerosis therapy: A cross-sectional pilot study.

Background: Teriflunomide is administered orally to treat relapsing-remitting multiple sclerosis. In this prospective pilot study, the free and total serum concentrations of teriflunomide obtained during routine health care were measured and their relationship with disease activity was evaluated.

Methods: Eighty-nine patients were included in this study.

Liquid chromatography-tandem mass spectrometry method for determination of total and free teriflunomide concentration in serum of patients with multiple sclerosis.

Teriflunomide belongs to disease-modifying drugs and is used in treatment of multiple sclerosis. According to in vitro studies more than 99.4 % of drug is binding to plasma proteins and only less than 1 % is free for clinical activity.

Effect of Blood Component Coatings of Enosseal Implants on Proliferation and Synthetic Activity of Human Osteoblasts and Cytokine Production of Peripheral Blood Mononuclear Cells.

The study monitored in vitro early response of connective tissue cells and immunocompetent cells to enosseal implant materials coated by different blood components (serum, activated plasma, and plasma/platelets) to evaluate human osteoblast proliferation and synthetic activity and inflammatory response presented as a cytokine profile of peripheral blood mononuclear cells (PBMCs) under conditions imitating the situation upon implantation. The cells were cultivated on coated Ti-plasma-sprayed (Ti-PS), Ti-etched (Ti-Etch), Ti-hydroxyapatite (Ti-HA), and ZrO2 surfaces. The plasma/platelets coating supported osteoblast proliferation only on osteoconductive Ti-HA and Ti-Etch whereas activated plasma enhanced proliferation on all surfaces.

The level of fatty acid-binding protein 4, a novel adipokine, is increased in rheumatoid arthritis and correlates with serum cholesterol levels.

Objective: To assess the expression of the novel adipokine Fatty Acid Binding Protein-4 (FABP4) in synovial tissues, serum and the synovial fluid of patients with rheumatoid arthritis (RA) and osteoarthritis (OA) and to study the relationships among FABP4, disease activity and metabolic status.

Methods: FABP4 levels were measured in the serum and synovial fluid of 40 patients with RA and 40 control patients with OA. The disease activity score (DAS28), C-reactive protein (CRP) levels and serum lipids were assessed in patients with RA.

The interaction of osteoblasts with bone-implant materials: 1. The effect of physicochemical surface properties of implant materials.

This comparative study of various surface treatments of commercially available implant materials is intended as guidance for orientation among particular surface treatment methods in term of the cell reaction of normal human osteoblasts and blood coagulation. The influence of physicochemical surface parameters such as roughness, surface free energy and wettability on the response of human osteoblasts in the immediate vicinity of implants and on the blood coagulation was studied. The osteoblast proliferation was monitored and the expression of tissue mediators (TNF-alpha, IL-8, MMP-1, bone alkaline phosphatase, VCAM-1, TGF-beta) was evaluated after the cell cultivation onto a wide range of commercially available materials (titanium and Ti6Al4V alloy with various surface treatments, CrCoMo alloy, zirconium oxide ceramics, polyethylene and carbon/carbon composite).

Clinical correlations of potential activity markers in systemic sclerosis.

The objectives of the project were the following: (1) to establish a group of patients with a confirmed diagnosis of systemic sclerosis (Ssc), (2) to perform a detailed entrance examination of each patient, (3) to determine concentrations of potential activity markers, and (4) to make a comprehensive examination of each patient 1 year after inclusion into the study. A total of 49 patients were examined, 36 with a limited form of SSc, 9 with diffuse SSc, and 4 with other forms of SSc. We determined plasma or serum levels of the N-terminal propeptide of procollagen type III (NPIIIP), interleukin-6 (IL-6), soluble receptor for interleukin-2 (sIL-2r), soluble intercellular adhesion molecule-1 (sICAM-1), soluble vascular adhesion molecule-1 (sVCAM-1), von Willebrand factor antigen (vWFAg), and big endothelin-1 (BET-1) using commercial kits, and urinary excretion of pyridinoline (PYR) and deoxypyridinoline (D-PYR) using high-performance liquid chromatography.

Biological properties of the intervertebral cages made of titanium containing a carbon-carbon composite covered with different polymers.

Intervertebral cages are used in orthopaedics for stabilization of injured lumbar parts of vertebral columns. Our study provides preliminary results of tests of the biological properties of titanium cages with a variously modified carbon/carbon composite (C/C) core. This core was produced from a C/C composite modified by hydrogel materials based on poly(2-hydroxyethyl methacrylate) (HEMA) enriched with 1% collagen or 35% methylmethacrylate or 30% terc-butylmethacrylamide.

Biological and biochemical properties of the carbon composite and polyethylene implant materials.

We studied the biocompatibility of the carbon composites and polyethylene materials with and without collagen or collagen and proteoglycan cover. We used the in vitro technology to study the adhesion of model cells evalution, their metabolic activity and the production of TNF-alpha as a cytokine model. Under in vivo condition, the biocompatibility of tested polymers were studied in the implantation experiment, subcutaneously in the interscapular region in the laboratory rat.

Biomechanical and biological properties of the implant material carbon-carbon composite covered with pyrolytic carbon.

The aim of this study was to test C/C material (carbonized, graphitized or covered with pyrolytic carbon) designated for the use in orthopaedic and bone surgery. Using an in vitro assay we confirmed, that the cell proliferation was exhibited the mostly on the C/C composite coated with pyrolitic carbon and afterwards polished. The two latest of subsequent water extracts of this material had a slightly inhibiting effect on the cells metabolic activity.

Autoantibodies can be prognostic markers of an erosive disease in early rheumatoid arthritis.

Objective: To evaluate a contribution of selected laboratory parameters for a prediction of progressive and erosive development in patients with early rheumatoid arthritis (RA).

Methods: In a prospective study baseline levels of antibodies to cyclic citrullinated peptide (anti-CCP), IgM, IgA, and IgG rheumatoid factors (RFs) were measured by enzyme linked immunosorbent assay (ELISA) in 104 patients with RA with disease duration <2 years. Antikeratin antibodies (AKA) and antiperinuclear factor (APF) were detected by indirect immunofluorescence.

[Relation between cytokines, adhesive immunoglobulins and matrix metalloproteinases in osteoarthritic joints].

Background: The dysbalance of proteinase production and their inhibitors leads to destruction of tissues in many pathological processes. Recently it was shown that the expression of proteinases is regulated also by interactions of cells which is mediated by adhesive molecules. We wanted to find out whether this mechanism is involved also in the destruction of joints in osteoarthritis.

[Use of Bioimplants to Replace Cartilage Part II: Application of Implants in Animal Experiments.].

The published paper pertains to a group of fullgrown minipigs where under anaesthesia with Narcamone after premedication with Rometar cartilage samples from the medial condyle of the femur in the area of the femoropatellar joint were taken. The collected cartilage samples served subsequently for the preparation of an autologous implant. After its preparation another operation was performed where by means of a bioptic trocar (diameter 3.

[Use of Bioimplants to Replace Cartilage Part I: Chondrocyte Cultivation in Three-dimensional Gel.].

For the investigation pig chondrocytes were used which were cultivated in a medium containing cartilaginous collagen and aggregan. The substances used were identified by electrophoresis, chromatography and electron microscopy. Various conditions for cultivating chondrocytes were tested - a) a medium containing cartilaginous collagens only, b) with addition of aggregan, c) with addition of the tripeptide GHK, d) with addition of the tripeptide and aggregan.

Effect of the tripeptide glycyl-L-histidyl-L-lysine on the proliferation and synthetic activity of chick embryo chondrocytes.

Under certain conditions chondrocytes form lattices with cartilage collagens, which may serve as cartilage implants. It is necessary to find the optimal conditions for culturing chondrocytes. Three different supports are compared: (a) plastic; (b) cartilage collagens; and (c) insoluble skin collagen solubilized under denaturing conditions (ISC-40).

[The effect of aluminum on the structure and metabolism of collagen].

Background: Aluminium is considered to be the etiopathogenetic factor in various pathological conditions. It was demonstrated already previously that metals even under conditions in vivo link with collagen structures and influence the protein metabolism. Therefore the authors investigated the effect of aluminium (Al) on collagen and its metabolism.

The effect of different collagens and of proteoglycan on the retraction of collagen lattice.

The effect of various collagens and proteoglycan on the formation and retraction of collagen lattices was tested. The most rapid aggregation of collagen molecules was observed by the use of the least cross-linked collagen fractions (ie pepsin-digested calf skin collagen type I). Lattices formed with more cross-linked collagens (acid soluble collagen-ASC, type III) contracted slowly and less intensively.

[The Influence of the Implanted Material (Glass Carbon) for the Proliferation of the Cell's.].

The authors have examined the influence of the implanted material (glass carbon) for the proliferation of human embryonal fibroblasts. It has been found that the tested material did not inhibit the cell's proliferation and it performed a good biocompatibility. Key words: fibroblasts, implanted material, glass carbon.

[Report on the influence of some materials on the proliferation of cells.].

The authors have examined the biocompatibility of four materials used for the production of implants with the proliferation of cells. This applies mainly to Ti-6 AI-4 V, Ti- 5AI-2,5 Fe, AKV-Ultra 2 alloys and also to the carbon filament connected by polyamide. In their study the authors used diploid clone -human embryonal fibroblasts LEP and heteroploid lines - Vero cells.

The influence of some non-steroidal anti-inflammatory drugs on the retraction of collagen lattices.

The effects of some antirheumatics on the formation and retraction of collagen lattices seeded with fibroblasts have been studied. Among the antirheumatics, diclofenac was the most active inhibitor of lattice retraction, then tropesin and to a lesser extent indomethacin. Ibuprofen which is known as a very slight inhibitor of protein synthesis was able to significantly enhance lattice retraction when 10 micrograms/ml (48.

Comparison of the influence of gelatine and collagen substrates on growth of chondrocytes.

The effect of culture substrates on adhesion and growth of chondrocytes and the influence of varying amounts of foetal calf serum on cell proliferation are compared. Skin and cartilage gelatines and type II collagen were used as substrates. The cells used in the experiments were the primary cultures of chondrocytes, frozen primary cultures of chondrocytes and chondrocytes after the 1st subculture.

Determination of radiolabelled proline and hydroxyproline in collagen hydrolysates by high-performance liquid chromatography with on-line radiometric detection.

Radiolabelled proline and hydroxyproline were separated on a C8 column (10 cm X 4.6 mm I.D.

Effect of tolfenamic acid on the metabolism of the main connective tissue components in rats.

Tolfenamic acid (Clotam) has been used in the therapy of rheumatic diseases for some years. Regarding its chemical structure it belongs to the group of fenamates. The effect of tolfenamic acid on the synthesis of collagen and proteoglycans in granulation tissue, skin or cartilage of rat weanlings was tested and compared with the action of mefenamic acid.