Evaluation of Mechanical Properties and Filler Interaction in the Field of SLA Polymeric Additive Manufacturing.

The paper deals with research focused on the use of fillers in the field of polymeric materials produced by additive technology SLA (stereolithography). The aim of the research is to evaluate 3D printing parameters, the mechanical properties (tensile strength, hardness), and the interaction of individual phases (polymer matrix and filler) in composite materials using SEM analysis. The tested fillers were cotton flakes and ground carbon fibres in different proportions. For the photosensitive resins, the use of cotton flakes as filler was found to have a positive effect on the mechanical properties not only under static but also under cyclic loading, which is a common cause of material failure in practice. The cyclic stress reference value was set at an amplitude of 5-50% of the maximum force required to break the pure resin in a static tensile test. A positive effect of fillers on the cyclic stress life of materials was demonstrated. The service life of pure resin was only 168 ± 29 cycles. The service life of materials with fillers increased to approximately 400 to 540 cycles for carbon fibre-based fillers and nearly 1000 cycles for cotton flake-based fillers, respectively. In this paper, new composite materials suitable for the use of SLA additive manufacturing techniques are presented. Research demonstrated the possibilities of adding cotton-based fillers in low-cost, commercially available resins. Furthermore, the importance of material research under cyclic loading was demonstrated.

Development of chronic venous disease.

INTRODUCTION: Chronic venous disease is known to gradually worsen in the course of years. However, little clinical data is available. The patient group and methods: We used the set of our patients with completed clinical examination, duplex sonography and photoplethysmography and selected 160 patients examined at our office after 10 and more years. Females represented 79% in the group. The mean age was 54 years (range 18-82 years). Venous varices in lower limbs in the family were reported by 49% patients. Venous sclerotization had been undergone by 6.3% and venous surgery by 10.6% patients. The mean body mass index was 27. We used CEAP classification. The clinical class C1 included 50 patients, C2 included 81, C3 included 15 and C4 included 14 patients. Regurgitation in superficial veins was found in 43%. The mean venous return time after physical activity was 24.5 s. RESULTS: The last examination was performed after 13 years on average (range 10-28 years). Deterioration of the clinical condition and progression to a higher class were observed in 36% patients in class C1, in 23.5% patients in C2, in 13% patients in C3 and in 7% patients in C4. We did not demonstrate any statistically significant effect of the followed parameters on the progression of the disease. CONCLUSION: The progression rate of the disease in our patient group followed at the vascular office for 10 and more years is lower compared to population studies.

Research on Low-Cycle Fatigue Engineered Hybrid Sandwich Ski Construction.

This research is aimed at evaluating the effect of low-cycle fatigue on a newly designed hybrid sandwich ski structure to determine the changes that may occur due to cyclic loading and thus affect its use. This is primarily concerned with the fatigue behavior of the tested ski over different time intervals simulating its seasonal use and its effect on the mechanical properties of the ski, i.e., the durability and integrity of the individual layers of the sandwich ski structure. The ski was subjected to 70,000 deflections by moving the crossbar by 60 mm according to the ski deflection calculation in the arch. The results of the cyclic tests of the engineered ski design showed no significant changes in the ski during loading. The average force required to achieve deflection in the first 10,000 cycles was 514.0 ± 4.2 N. Thereafter, a secondary hardening of the structure occurred during relaxation and the force required increased slightly to 543.6 ± 1.7 N. The required force fluctuated slightly during the measurements and in the last series the value was 540.4 ± 0.8 N. Low-cycle fatigue did not have a significant effect on the mechanical properties of the ski; there was no change in shape or visual delamination of the individual layers of the structure. From the cross-section, local delamination was demonstrated by image analysis, especially between the Wood core and the composite layers E-Glass biaxial and Carbon triaxial.

Chemical and Structural Analysis of Newly Prepared Co-W-Al Alloy by Aluminothermic Reaction.

This article is devoted to the characterization of a new Co-W-Al alloy prepared by an aluminothermic reaction. This alloy is used for the subsequent preparation of a special composite nanopowder and for the surface coating of aluminum, magnesium, or iron alloys. Due to the very high temperature (2000 °C-3000 °C) required for the reaction, thermite was added to the mixture. Pulverized coal was also added in order to obtain the appropriate metal carbides (Co, W, Ti), which increase hardness, resistance to abrasion, and the corrosion of the coating and have good high temperature properties. The phase composition of the alloy prepared by the aluminothermic reaction showed mainly cobalt, tungsten, and aluminum, as well as small amounts of iron, titanium, and calcium. No carbon was identified using this method. The microstructure of this alloy is characterized by a cobalt matrix with smaller regular and irregular carbide particles doped by aluminum.

Ultrasonic Pulsating Water Jet Peening: Influence of Pressure and Pattern Strategy.

Peening techniques are nowadays attracting more research attention due to their association with the extending of the service life and improving surface texture of engineering components. Ultrasonic pulsating water jet peening represents a new way of mechanical surface treatment. Accelerated water droplets via hammer effect cause small elastic-plastic deformations on the surface. This work deals with peening of aluminum alloy using an ultrasonic pulsating water jet, where periodically acting water droplets were used as the peening medium. The aim of the work was the feasibility study of the peening process and to observe the effects of pressure (p = 10, 20 and 30 MPa) and pattern trajectory (linear hatch and cross hatch). The peened surfaces were analyzed by the surface roughness profile parameters Ra and Rz and the microhardness along the peening axis into the material. Graphically processed results show a clear increase of measured values with increasing pressure (p = 10, 20 and 30 MPa), where the roughness values ranged from 1.89 µm to 4.11 µm, and the microhardness values ranged from 43.3 HV0.005 to 47 HV0.005, as compared to 40.3 HV0.005 obtained for the untreated sample. The achieved results indicate potential using of an ultrasonic pulsating water jet as a new method of surface treatment of metals. By controlled distribution of water droplets, it is possible to achieve a local distribution of surface roughness, and at the same time, strengthening of the subsurface layers in the material without thermal influence on the material.

Does the time from symptoms onset to treatment impact further development of vein thrombosis in the leg?

INTRODUCTION: It is unknown how many days pass from the symptom onset to anticoagulation therapy of vein thrombosis (VT) in the leg in our country and whether this time has an impact on further development of the disease. METHODS: Among patients treated at our department in recent years for deep vein thrombosis in the leg, we selected 50 patients with the required data available. In each case the diagnosis was confirmed by duplex sonography. The mean age was 51 years and the group included 31 males and 19 females. Proximal VT was found in 72 % of the patients. Idiopathic VT was found in 38 % of the patients. The patients were followed for 3-6 months. RESULTS: The mean time to initiation of VT therapy was 10 days, range 1-42 days, median 7 days. Two thirds of the patients were treated with novel oral anticoagulants. Residual VT (incomplete recanalization) was present after 1-2 months of the therapy in 40.9 %, and after 3-6 months in 25 % of the patients. The positive relationship between the time to treatment and residual VT was significant both after 1-2 months of therapy (p < 0.05) and after 3-6 months of therapy (p < 0.01). CONCLUSION: The time to VT treatment is clearly longer in our country compared to other countries. Delayed onset of treatment reduces the success rate of the therapy as assessed by an increased risk of residual thrombosis in the affected veins in subsequent months. Key words: delayed diagnosis - residual vein thrombosis - vein thrombosis.

Bacillus subtilis alters the proportion of major membrane phospholipids in response to surfactin exposure.

Surfactin, an anionic lipopeptide produced by Bacillus subtilis, is an antimicrobial that targets the cytoplasmic membrane. Nowadays it appears increasingly apparent that the mechanism of resistance against these types of antibiotics consists of target site modification. This prompted us to investigate whether the surfactin non-producing strain B. subtilis 168 changes its membrane composition in response to a sublethal surfactin concentration. Here we show that the exposure of B. subtilis to surfactin at concentrations of 350 and 650 µg/ml (designated as SF350 and SF650, respectively) leads to a concentration-dependent growth arrest followed by regrowth with an altered growth rate. Analysis of the membrane lipid composition revealed modifications both in the polar head group and the fatty acid region. The presence of either surfactin concentration resulted in a reduction in the content of the major membrane phospholipid phosphatidylglycerol (PG) and increase in phosphatidylethanolamine (PE), which was accompanied by elevated levels of phosphatidic acid (PA) in SF350 cultures. The fatty acid analysis of SF350 cells showed a marked increase in non-branched high-melting fatty acids, which lowered the fluidity of the membrane interior measured as the steady-state fluorescence anisotropy of DPH. The liposome leakage of carboxyfluorescein-loaded vesicles resembling the phospholipid composition of surfactin-adapted cells showed that the susceptibility to surfactin-induced leakage is strongly reduced when the PG/PE ratio decreases and/or PA is included in the target bilayer. We concluded that the modifications of the phospholipid content of B. subtilis cells might provide a self-tolerance of the membrane active surfactin.

Mimicking of Arginine by Functionalized N(ω)-Carbamoylated Arginine As a New Broadly Applicable Approach to Labeled Bioactive Peptides: High Affinity Angiotensin, Neuropeptide Y, Neuropeptide FF, and Neurotensin Receptor Ligands As Examples.

Derivatization of biologically active peptides by conjugation with fluorophores or radionuclide-bearing moieties is an effective and commonly used approach to prepare molecular tools and diagnostic agents. Whereas lysine, cysteine, and N-terminal amino acids have been mostly used for peptide conjugation, we describe a new, widely applicable approach to peptide conjugation based on the nonclassical bioisosteric replacement of the guanidine group in arginine by a functionalized carbamoylguanidine moiety. Four arginine-containing peptide receptor ligands (angiotensin II, neurotensin(8-13), an analogue of the C-terminal pentapeptide of neuropeptide Y, and a neuropeptide FF analogue) were subject of this proof-of-concept study. The N(ω)-carbamoylated arginines, bearing spacers with a terminal amino group, were incorporated into the peptides by standard Fmoc solid phase peptide synthesis. The synthesized chemically stable peptide derivatives showed high receptor affinities with Ki values in the low nanomolar range, even when bulky fluorophores had been attached. Two new tritiated tracers for angiotensin and neurotensin receptors are described.

Surfactin production enhances the level of cardiolipin in the cytoplasmic membrane of Bacillus subtilis.

Surfactin is a cyclic lipopeptide antibiotic that disturbs the integrity of the cytoplasmic membrane. In this study, the role of membrane lipids in the adaptation and possible surfactin tolerance of the surfactin producer Bacillus subtilis ATCC 21332 was investigated. During a 1-day cultivation, the phospholipids of the cell membrane were analyzed at the selected time points, which covered both the early and late stationary phases of growth, when surfactin concentration in the medium gradually rose from 2 to 84µmol·l(-1). During this time period, the phospholipid composition of the surfactin producer's membrane (Sf(+)) was compared to that of its non-producing mutant (Sf(-)). Substantial modifications of the polar head group region in response to the presence of surfactin were found, while the fatty acid content remained unaffected. Simultaneously with surfactin production, a progressive accumulation up to 22% of the stress phospholipid cardiolipin was determined in the Sf(+) membrane, whereas the proportion of phosphatidylethanolamine remained constant. At 24h, cardiolipin was found to be the second major phospholipid of the membrane. In parallel, the Laurdan generalized polarization reported an increasing rigidity of the lipid bilayer. We concluded that an enhanced level of cardiolipin is responsible for the membrane rigidification that hinders the fluidizing effect of surfactin. At the same time cardiolipin, due to its negative charge, may also prevent the surfactin-membrane interaction or surfactin pore formation activity.

Microchip-ESI-MS determination of dissociation constant of the lysozyme-NAG(3) complex.

An on-line microfluidic system for determination of dissociation constants of enzyme/substrate weak interactions by nanoESI-MS is introduced. The microchip was designed to permit the enzyme and the substrate to mix by molecular diffusion in a pressure-driven laminar flow. Introduction of reagent solutions into the chip was an optimized combination of a micropump and an autosampler to enable automation of the measurements. The system performance was tested for monitoring of non-covalent interactions of hen egg white lysozyme with N-acetyl glucosamine oligomers. Dissociation constant (K(D)) of the hen egg white lysozyme-N-acetyl glucosamine oligomer complex was determined by non-linear regression analysis, and the range of K(D) values (39+/-6 x 10(-6) and 19.6+/-8 x 10(-6) M for manual and automated infusions, respectively) confirms the previously reported values. Such miniaturization of a continuous-flow enzyme assay system to a microfluidic format can maximize the capabilities of mass spectrometric detection, reduce the sample size and analysis time required, as well as the associated costs for on-line enzymatic kinetic studies.

Long-term adaptation of Bacillus subtilis 168 to extreme pH affects chemical and physical properties of the cellular membrane.

We characterized physical and chemical properties of cell-membrane fragments from Bacillus subtilis 168 (trpC2) grown at pH 5.0, 7.0 and 8.5. Effects of long-term bacterial adaptation reflected in growth rates and in changes of the membrane lipid composition were correlated with lipid order and dynamics using time-resolved fluorescence anisotropy of 1,6-diphenyl-1,3,5-hexatriene. We demonstrate that the pH adaptation results in a modification of a fatty acid content of cellular membranes that significantly influences both the lipid-chain order and dynamics. For cultivation at acidic conditions, the lipid order increases and membrane dynamics decreases compared to pH 7.0. This results in rigid and ordered membranes. Cultivation at pH 8.5 causes slight membrane disordering. Instant pH changes induce qualitatively similar but smaller effects. Proton flux measurements performed on intact cells adapted to both pH 5.0 and 8.5 revealed lower cell-membrane permeability compared to bacteria cultivated at pH optimum. Our results indicate that both acidic and alkalic pH stress represent a permanent challenge for B. subtilis to keep a functional membrane state. The documented adaptation-induced adjustments of membrane properties could be an important part of mechanisms maintaining an optimal intracellular pH at a wide range of extracellular proton concentrations.

Diabetes and distal access location are associated with higher wall shear rate in feeding artery of PTFE grafts.

BACKGROUND: Surgical creation of permanent vascular access for haemodialysis leads to considerable haemodynamic changes. They could be implicated in the pathogenesis of access complications, which limit access survival, especially in diabetics. Physiologically, the relation between arterial diameter and blood velocity is maintained by wall shear stress (WSS), which is directly related to both blood viscosity and wall shear rate (WSR = blood velocity/internal diameter). Because of methodological difficulties, WSR is used as a measure of WSS. Extremely high values of WSS might induce hypercoagulable states, which might contribute to access thrombosis. We performed a study, which was aimed to (i) describe WSR values in feeding arteries of various polytetrafluoroethylene access types and (ii) prove that diabetic patients have higher WSR than non-diabetics. METHODS: A linear-array 11 MHz probe of SONOS 5500 (Phillips, USA) was used to obtain blood velocity and internal diameter in the feeding arteries of radial or brachial polytetrafluoroethylene grafts. WSR was calculated as 4 x blood velocity/internal diameter. We compared observed values of WSR according to feeding artery (radial vs brachial artery) and according to diabetic status using unpaired t-test. RESULTS: We included 106 patients (58 non-diabetic and 48 diabetic) in the study. WSR was significantly higher in radial arteries compared with brachial arteries independent of diabetes status. Diabetic subjects had significantly higher WSR in both radial and brachial arteries. CONCLUSIONS: Diabetes mellitus and distal vascular access creation are associated with higher WSR in the feeding artery. This could be of relevance in the pathogenesis of access complications, e.g. thrombosis, and thus lower patency rates in diabetic patients.

Pulse wave analysis during supine rest may identify subjects with recurrent vasovagal syncope.

In the present study, we studied whether analysis of the FAP (finger arterial pressure) waveform during supine rest discriminates subjects with recurrent VVS (vasovagal syncope) from healthy controls. Signal-averaged FAP waveforms (Finapres) were obtained in 32 head-up tilt-test-positive subjects with recurrent VVS (35 +/- 13 years) and in 32 sex- and age-matched healthy controls. The DT (time delay) between the systolic and diastolic peaks of the FAP waveform was measured and large artery SI (stiffness index) was calculated as a ratio of body height and DT. VVS patients had significantly shorter DT compared with controls (303 +/- 31 compared with 329 +/- 18 ms; P < 0.001) and higher SI (5.79 +/- 0.70 compared with 5.20 +/- 0.36 m/s; P < 0.001). The differences were independent of heart rate and blood pressure. SI > 5.45 m/s identified subjects with syncope with a sensitivity of 72% and a specificity of 84%. Age-corrected DT (cDT = DT + age-350) identified subjects with syncope with a sensitivity of 75% and a specificity of 84%. Combined use of cDT <0 ms and SI > 5.45 m/s increased sensitivity and specificity to 81% and 96% respectively. The discriminative power of FAP descriptors improved further when younger subjects were excluded. In subjects aged >30 years (median age), the combination of cDT and SI identified subjects with syncope with a sensitivity of 93% and a specificity of 100%. These results suggest that FAP descriptors during supine rest might be useful in the diagnosis of VVS in middle-aged subjects.

Regular ultrasonographic screening significantly prolongs patency of PTFE grafts.

BACKGROUND: Polytetrafluoroethylene (PTFE) dialysis grafts have considerably shorter patency than native arteriovenous fistulas, despite the use of a complex of screening monitoring methods (venous pressure, access flow). PTFE grafts are used often in subjects with depleted subcutaneous veins after previous abandoned accesses, so keeping the access patent is crucial. We hypothesized that regular duplex Doppler ultrasound screening for access stenoses, together with their sooner treatment, would prolong PTFE graft patency. METHODS: We performed a randomized, prospective study of PTFE grafts' cumulative patency in 192 subjects. In group 1, regular ultrasound examinations performed every 3 months was added to traditional screening (i.e., regular access examination at hemodialysis unit, monitoring of venous pressure and access flow). Group 2 was screened only traditionally (without ultrasound). Interventions of suspected stenoses were indicated by nephrologists, vascular surgeon, and, in group 1, also by ultrasonography. Classic ultrasound criteria for significant stenosis were used, even if the access flow had not been decreased. The mean follow-up lasted 392 +/- 430 days. RESULTS: Groups were similar with respect to age, gender, diabetes status, and number of previous abandoned accesses. Group 1 had significantly longer access patency (P < 0.001). Number of interventions per graft was 2.1 +/- 1.8 and 1.3 +/- 1.0 in group 1 and group 2. CONCLUSIONS: Regular screening duplex Doppler ultrasonography results in significantly longer PTFE graft patency due to early detection of access stenosis and, thus, more frequent elective interventions of access stenoses.

Different structural requirements for adenylate cyclase toxin interactions with erythrocyte and liposome membranes.

The bifunctional Bordetella adenylate cyclase toxin-hemolysin (ACT) penetrates target cell membranes, forms cation-selective channels and subverts cellular signaling by catalyzing uncontrolled conversion of ATP to cAMP. While primarily targeting phagocytes expressing the alphaMbeta2 integrin (CD11b/CD18), the toxin can also penetrate mammalian erythrocytes lacking the receptor and membrane endocytosis. We sought here to analyze the membrane interactions of ACT in a liposome model. Insertion of ACT into liposome membranes required calcium and caused leakage of entrapped fluorescent probes due to liposome disruption, as indicated by similar release kinetics for the approximately 398 Da FITC probe and its approximately 4400 Da dextran conjugate. However, the non-acylated proACT, which does not penetrate cellular membranes, exhibited higher capacity to bind and lyze liposomes than the mature toxin, showing that the fatty-acyl modification was not required for penetration of ACT into the lipid bilayer. Individual deletions within the channel-forming, acylation and repeat domains of ACT abolished its capacity to disrupt both liposomes and erythrocytes. In contrast to erythrocyte binding, however, the liposome binding was only lost upon a simultaneous deletion of both the channel-forming and acylation domains, suggesting that the acylation domain was also involved in liposome penetration of ACT. Moreover, substitutions of glutamates 509 and 516 by lysines, which strongly enhanced the channel-forming and hemolytic activity of ACT, did not affect its capacity to disrupt liposomes. This shows that the mechanism of ACT action in cellular membranes is not fully reproduced in liposome membranes.