TNF-alpha binding ability as a sign of peripheral blood neutrophils preactivation in the course of multiple sclerosis

Usually neglected is the role of neutrophils in causing of immunological disturbances in patients with multiple sclerosis (MS). Nevertheless, it has been indicated over the recent years that these cells possess a sufficient potential to affect both immune response and inflammation. This potential may result in MS through the process of priming of these cells by proinflammatory cytokines like TNF. We studied TNF and its soluble receptors sp55 and sp75 serum levels and binding of fluorescein isothiocyanate (FITC)-stained TNF by neutrophils. We studied three different groups of MS patients: 10 patients in relapse of the disease, 13 in its remission, and 11 in its chronic progressive form (CP-MS). The control was provided by 14 neurological patients (OND) with non-inflammatory diseases. The performed studies showed higher TNF sp55 and sp75 soluble TNF receptors serum levels in the patients with relapse, comparing with other MS patients and OND. TNF binding by neutrophils of MS patients during relapse was also higher, than other MS patients and OND. These result suggest the preactivation of neutrophils in the relapse of MS.     

Increased oxytocinergic system activity in the cardiac muscle in spontaneously hypertensive SHR rats

IntroductionThe present study aimed to determine whether the presence of cardiac hypertrophy due to arterial hypertension is associated with a change in the activity of the oxytocinergic system in cardiomyocytes.Material and methodsThe experiments were performed on male, spontaneously hypertensive rats (SHR, n = 10) and normotensive Wistar-Kyoto rats (WKY, n = 12). Blood samples were collected from both SHR and WKY animals to asses plasma oxytocin (OT) concentration; the rats were sacrificed by decapitation. Samples of the left and right ventricles were harvested for the analysis of the OT and oxytocin receptor (OTR) protein by ELISA, and OT and OTR mRNA expression by RT-PCR. Immunohistopathological studies were performed to confirm the presence of OTR receptors in the cardiac muscle of the ventricles.ResultsPlasma OT concentration did not differ between SHR and WKY rats. In the SHR rats, the expression of OT mRNA and the OT protein level was higher in the left and the right ventricle, while OTR mRNA expression was significantly lower in both the left and the right ventricle. However, the level of OTR protein was higher only in the left ventricle of the SHR rats. The presence of OTR receptors was confirmed by immunohistochemical analysis in the muscle of the right and left ventricle.ConclusionsThe presence of arterial hypertension is associated with increased activity of the oxytocinergic system in the heart, especially in the area of the left ventricle. These findings support the important role of this system in the maintenance of cardiovascular homeostasis.     

Phase Structure Evolution of the Fe-Al Arc-Sprayed Coating Stimulated by Annealing

The article presents the results of research on the structural evolution of the composite Fe-Al-based coating deposited by arc spray with initial low participation of in situ intermetallic phases. The arc spraying process was carried out by simultaneously melting two different electrode wires, aluminum and low alloy steel (98.6 wt.% of Fe). The aim of the research was to reach protective coatings with a composite structure consisting of a significant participation of Fe_xAl_y as intermetallic phases reinforcement. Initially, synthesis of intermetallic phases took place in situ during the spraying process. In the next step, participation of Fe_xAl_y fraction was increased through the annealing process, with three temperature values, 700 °C, 800 °C, and 900 °C. Phase structure evolution of the Fe-Al arc-sprayed coating, stimulated by annealing, has been described by means of SEM images taken with a QBSD backscattered electron detector and by XRD and conversion electron Mössbauer spectroscopy (CEMS) investigations. Microhardness distribution of the investigated annealed coatings has been presented.     

Mimicking the extracellular matrix with functionalized,metal-assembled collagen peptide scaffolds

Natural and synthetic three-dimensional (3-D) scaffolds that mimic the microenvironment of the extracellular matrix (ECM), with growth factor storage/release and the display of cell adhesion signals, offer numerous advantages for regenerative medicine and in vitro morphogenesis and oncogenesis modeling. Here we report the design of collagen mimetic peptides (CMPs) that assemble into a highly crosslinked 3-D matrix in response to metal ion stimuli, that may be functionalized with His-tagged cargoes, such as green fluorescent protein (GFP-His₈) and human epidermal growth factor (hEGF-His₆). The bound hEGF-His₆ was found to gradually release from the matrix in vitro and induce cell proliferation in the EGF-dependent cell line MCF10A. The additional incorporation of a cell adhesion sequence (RGDS) at the N-terminus of the CMP creates an environment that facilitated the organization of matrix-encapsulated MCF10A cells into spheroid structures, thus mimicking the ECM environment.     

Quantitative imaging of proteoglycan in cartilage using a gadolinium probe and microCT

OBJECTIVE: Micro-computed tomography (microCT) imaging has the potential to allow the three-dimensional (3D) visualization of cartilage morphology. However, cartilage intensity on a microCT image is weak because cartilage does not strongly attenuate X-rays. This work was designed to demonstrate that exposure of cartilage to charged gadolinium compounds modifies the intensity to allow an improved visualization of cartilage morphology and the determination of proteoglycan content. DESIGN: Trypsin was used to deplete proteoglycan in bovine nasal cartilage disks. Disks were then exposed to Gd(3+), gadopentetate (Gd-DTPA(2-)), or gadoteridol (Gd-HP-DO3A), and imaged with microCT. The intensities of the disks were measured from the images and compared to the actual proteoglycan content determined with a dimethylmethylene blue assay. RESULTS: Treatment of na?ve disks with 200 mM Gd(3+) for 24h at room temperature produced a 2.8-fold increase in intensity on microCT images. Similar treatment with 200 mM Gd-DTPA(2-) produced a 1.4-fold increase. After 2h of trypsin treatment at room temperature, the intensities of cartilage disks exposed to 20 0mM Gd(3+) decreased by 12%. Conversely, the intensities of trypsin-treated disks exposed to 200 mM Gd-DPTA(2-) increased by 15%. Trypsin treatment caused a 4% increase in the intensities of disks exposed to neutral Gd-HP-DO3A. The correlation between proteoglycan content and the microCT intensity of cartilage treated with Gd(3+) was very good (r(2)=0.81). CONCLUSIONS: Gadolinium and microCT allow an improved 3D visualization of cartilage and quantification of its proteoglycan content.     

Failures of open reduction in developmental dislocation of the hip

Eight patients with the same number of developmental dislocated hips were initially treated with open reduction through an anterolateral approach. The hips then became dislocated again. There were six girls, with a mean age at first open reduction of 13.3 months. We treated the hips with a new open reduction through an anteromedial approach. A constricted anteromedial capsule was always found as the main factor; all had an intact anteromedial capsule, there was an inverted transverse ligament in three cases and a very tight psoas tendon in another three cases. All were reduced, without complications and with only one simultaneous bone procedure. Risk of avascular necrosis and residual dysplasia could not be evaluated with this follow-up. We conclude that in any open reduction for developmental dislocation of the hip the surgeon must consider a release of the anteromedial capsule, which we have found to be the most important factor in technical failures.