The aim of this study was to analyse and identify specific buffalo seminal plasma proteins (SPPs) responsible for sperm cryotolerance during low temperature storage. Computer Assisted Sperm Analysis (CASA) of the motility and viability of buffalo spermatozoa was performed before freezing and after thawing. Two sample groups were formed – ejaculates with high cryotol- erance (group A) and low cryotolerance (group B). CASA demonstrated that the initial progres- sive motility after thawing of the spermatozoa in group A is significantly higher than in group B (p<0.001). Group B showed a significant increase in the percentage of static and non-progressive spermatozoa at 240 min, when compared to group A (p<0.05). SPPs, proteins in the cryoprotec- tive medium (PM) and proteins in the mixture of PM and SP were separated by High Perfor- mance Liquid Chromatography (HPLC). Comparative analysis of the chromatographic profiles was performed to identify specific proteins related to sperm cryotolerance. SPPs profiles showed 5 distinct protein peaks in both groups, ranging from 500 kDa to 50 Da. Chromatograms of group A and group B showed quantitative and qualitative differences in protein content. Chromato- grams of proteins in PM showed 11 well-expressed peaks. HPLC analysis of the mixtures of SPPs from the two groups and PM visualized the formation of a new bio-complex structure expressed by a protein peak specific for group A (7.674 min, AU 1.50). This protein peak can be referred as a phenotypic trait for buffalo ejaculates with high sperm cryotolerance.
The aim of this study was to identify the proteoforms of albumin and kallikrein in stallion seminal plasma (SP), and to determine their correlations with sperm motility parameters. The experimental material consisted of ejaculates from 8 stallions, which were collected during the breeding and non-breeding seasons (BS and NBS, respectively). SP proteins were identified by 2-D PAGE and mass spectrometry (MALDI TOT/TOF MS). Sperm motility parameters were analyzed using the CASA system. Protein expression (integrated optical density-IOD) of albumin proteoforms 1 (ALB 1) and 2 (ALB 2) and kallikrein proteoforms 1 (KAL 1) and 2 (KAL 2) was correlated (p<0.05) with sperm motility parameters (total motility and progressive motility) during the BS. No significant correlations were found between the expression of albumin or kallikrein and sperm motility parameters during the NBS. The presence of correlations between the expression of ALB 1, ALB 2, KAL 1, KAL 2 and selected sperm motility parameters could suggest that the analyzed components of the SP belong to the group of fertility-associated pro- teins (FAPs).
In our recent study we demonstrated that the holding of fresh semen in fractionated seminal plasma (SP1, >40 kDa; SP2, <40 kDa), obtained by gel filtration chromatography, significantly improved the sperm quality characteristics following cryopreservation (Wasilewska-Sakowska et al. 2019). In this study we investigated the effect of post-thaw (PT) supplementation of fractionated SP (SP1 and SP2) on the survival of spermatozoa from boars with good and poor semen freezability, GSF and PSF, respectively. Two-dimensional polyacrylamide gel electrophoresis (2D-PAGE) analysis showed distinct differences in the protein profiles of SP1 and SP2 from boars with GSF or PSF regarding the number of protein spots. Sperm motility characteristics and the motion patterns, assessed using the computer-assisted sperm analysis (CASA) system, were markedly higher in PT semen supplemented with SP1 and SP2 from boars with GSF. Post-thaw supplementation of either SP1 or SP2 from boars with GSF significantly improved mitochondrial function, plasma membrane and acrosome integrity, and viability during storage. The findings of this study have confirmed that the presence of protective protein components in varying abundance in either fractionated SP from boars with good freezability ejaculates significantly improved the sperm survival following PT storage.
Introduction: Platelet-rich plasma (PRP) preparations can be used in bone tissue healing but there are numerous doubts among clinical orthopedists about effectiveness of this method. Materials and methods: The studies were carried out in 12 rabbits of white termond breed. In operating room we operationally generated cylindrical, unicortical defects of the diameter of 4 mm in the middle of the shafts of both femurs. The defects in the left bones were left without filling and served as controls, and 0.7 ml of the ready-to-use PRP was administered to the defects in the right bones (experimental group). We evaluated the usefulness of the diagnostic methods applied: biomechanical tests, micro-CT tests, densitometry, typical radiology, macroscopic measurements, histopathological examinations. Results: The macroscopic measurements showed a statistically significant increase in the dimension in the area of the right defect filled with PRP in relation to the control group. In experimented group, the assessment of the X-ray images showed the formation of a callus cuff around the defects. Densitometric examinations showed no statistically significant differences between defects in the experimental and control group. The analysis of the micro-CT examina- tions showed an increase in the total volume of the tissue examined (Vb) and the low density tissue fraction (Vb2) in the experimental group. The biomechanical examinations revealed signi- ficant decrease in the maximum breaking force (F max) necessary to break the bone in the experi- mental group in relation to the control group. Conclusions: Platelet-rich plasma (PRP) stimulates bone formation in the area of bone defects and may accelerate bone regeneration.
This article deals with the testing of surface layers produced on technical titanium Ti99.2 under glow discharge conditions. In order to determine the effect of process temperature on the produced surface layers, nitriding processes were carried out at 700°C and 800°C and for 3 and 5 hours. The research results on evaluating the properties of the obtained surface layers and the characterization of their morphology were presented. The impact of the adopted nitriding process variant on the quality of the obtained layers was evaluated. It was demonstrated that the use of the supplementary potential during the ion nitriding process reduces the unwanted edge effect, which results in a significant increase in the homogeneity of the nitrided layers and improves the functional properties of the technical titanium Ti99.2.
This paper presents an analysis of use of ultrasonic standing wave in cell separation from bodily fluids based on the example of erythrocyte separation from plasma. It describes movement of red blood cells in plasma under the influence of the acoustic field (whose forces result from interaction of red blood cells with plasma as the vibrating medium) and under the influence of resistance forces in Stokes’ and Oseen’s approximation. The general properties of solutions of the motion equation are given. The solutions for the parameters of the ultrasonic wave and blood cells which are interesting in terms of practical applications in medical diagnostics are discussed. Time constants of the cell transportation to the regions of stable equilibrium in the field of ultrasonic standing wave are estimated. The formulas which determine the time needed to obtain the assumed concentration increase in plasma in nodes and/or anti-nodes of the standing wave are derived.
The purpose of the study was to assess the effect of autologous activated platelet-rich plasma on healing of a bone substitute - tricalcium phosphate hydroxyapatite in experimental long bone defects using an animal model. The experiment involved an animal model of femoral defect. 24 Termond white rabbits were used in the study. We evaluated the effect of autologous platelet-rich plasma on tricalcium phos- phate using classical radiology, micro-CT studies, strength tests and histological evaluation. Radiological and histological assessment did not show a beneficial effect of PRP together with a bone substitute in comparison to filling the defects only with bone replacement material. The only benefit of adding platelet-rich plasma to a bone substitute was shown in microCT imaging. Autologous, activated platelet-rich plasma combined with hydroxyapatite tricalcium phos- phate has a positive effect on the remodeling of the newly formed bone tissue, increasing its density.
Filtering nonwovens produced with melt-blown technology are one of the most basic materials used in the construction of respiratory protective equipment (RPE) against harmful aerosols, including bio- and nanoaerosols. The improvement of their filtering properties can be achieved by the development of quasi-permanent electric charge on the fibres. Usually corona discharge method is utilized for this purpose. In the presented study, it was assumed that the low-temperature plasma treatment could be applied as an alternative method for the manufacturing of conventional electret nonwovens for the RPE construction. Low temperature plasma treatment of polypropylene nonwovens was carried out with various process gases (argon, nitrogen, oxygen or air) in a wide range of process parameters (gas flow velocity, time of treatment and power supplied to the reactor electrodes). After the modification, nonwovens were evaluated in terms of filtration efficiency of paraffin oil mist. The stability of the modification results was tested after 12 months of storage and after conditioning at elevated temperature and relative humidity conditions. Moreover, scanning electron microscopy and ATR-IR spectroscopy were used to assess changes in surface topography and chemical composition of the fibres. The modification of melt-blown nonwovens with nitrogen, oxygen and air plasma did not result in a satisfactory improvement of the filtration efficiency. In case of argon plasma treatment, up to 82% increase of filtration efficiency of paraffin oil mist was observed in relation to untreated samples. This effect was stable after 12 months of storage in normal conditions and after thermal conditioning in (70 ± 3)°C for 24 h. The use of low-temperature plasma treatment was proven to be a promising improvement direction of filtering properties of nonwovens used for the protection of respiratory tract against harmful aerosols.
This work reports the results of a study of Mo thin films synthesis by DC Pulsed Magnetron Sputtering method (PMS), operating at pulse main frequency of 100 kHz and modulated by the additional modulation frequency, driving in the range of 5-1000 Hz (modulated Pulse Magnetron Sputtering – mPMS). We have studied the influence of mPMS on plasma chemical reactions and mechanisms of layer growth using optical emission spectroscopy technique. Our experiment showed strong influence of mPMS method, on the morphology (scanning electron microscopy), phase composition (X-ray diffractometry) and electric properties (4-point probes method) of nanocrystalline and amorphous Mo films. From the utilitarian point of view, low value of resistivity – 43,2 μΩcm of synthesized Mo films predestines them as back contacts for thin solar cells CIGS. Our results revealed that additional modulation frequency should be considered as an important factor for optimization of films synthesis by means of PMS-based methods.
In this work, the change of the structure and microhardness of Ti6Al4V titanium alloy after remelting and remelting with SiC alloing by electric arc welding (GTAW method) was studied. The current intensity equal 100 A and fixed scan speed rate equal 0,2 m/min has been used to remelting surface of the alloy. Change of structure were investigated by optical and scanning electron microscopy. Microhardness test showed, that the remelting of the surface does not change the hardness of the alloy. Treated by GTAW SiC alloying leads to the formation of hard (570 HV0, 1) surface layer with a thickness of 2 mm. The resulting surface layer is characterized by diverse morphology alloyed zone. The fracture of alloy after conventional heat treatment, similarly to fracture after remelting with GTAW is characterized by extremely fine dimples of plastic deformation. In the alloyed specimens the intergranular and crystalline fracture was identified.
The mechanical behavior and the change of retained austenite of nanocrystalline Fe-Ni alloy have been investigated by considering the effect of various Ni addition amount. The nanocrystalline Fe-Ni alloy samples were rapidly fabricated by spark plasma sintering (SPS). The SPS is a well-known effective sintering process with an extremely short densification time not only to reach a theoretical density value but also to prevent a grain growth, which could result in a nanocrystalline structures. The effect of Ni addition on the compressive stress-strain behavior was analyzed. The variation of the volume fraction of retained austenite due to deformation was quantitatively measured by means of x-ray diffraction and microscope analyses. The strain-induced martensite transformation was observed in Fe-Ni alloy. The different amount of Ni influenced the rate of the strain-induced martensite transformation kinetics and resulted in the change of the work hardening during the compressive deformation.
In view of their advantageous properties (high hardness, good frictional wear resistance, chemical and thermal stability at elevated temperatures), cubic boron nitride (cBN) and tungsten carbide (WC) are commonly used for the fabrication of cutting tools. The composites were consolidated at a temperature of 1100°C under a load of 100 MPa for 10 min. The density of the thus produced material was close to the theoretical value (about 99.6%), and the hardness HV30 was about 1950. The phases identified in the composite were WC, Co, and cBN. Microstructural examinations revealed that numerous trans-crystalline fractures through the cBN particles occurred in the material. The present study is concerned with the wear of the WCCo and WCCo/cBN composites. Comparative tribological examinations were performed in a tribological tester using the ball-on-disc arrangement under the conditions of dry friction. The counterspecimens were steel and Al2O3 balls. The tests were conducted under a unit load of 10 N. After the tests, the surface of the samples was examined to describe the wear mechanisms active in various composite materials.
In this study, precisely controlled large scale gas atomization process was applied to produce spherical and uniform shaped high entropy alloy powder. The gas atomization process was carried out to fabricate CoCrFeNiMn alloy, which was studied for high ductility and mechanical properties at low temperatures. It was confirmed that the mass scale, single phase, equiatomic, and high purity spherical high entropy alloy powder was produced by gas atomization process. The powder was sintered by spark plasma sintering process with various sintering conditions, and mechanical properties were characterized. Through this research, we have developed a mass production process of high quality and spherical high entropy alloy powder, and it is expected to expand applications of this high entropy alloy into fields such as powder injection molding and 3D printing for complex shaped components.
In this study T6 heat treated 6063 aluminum alloys were used as substrate material. In order to form a bond between the substrate and the main coating, all samples were coated with Ni-Cr-Al powders. 8 wt% Yttria Stabilized Zirconia powders (YSZ) were coated with plasma spray technique. Thickness of YSZ was 150 m and bond coating was 36 m. XRD and SEM-EDS analyses were performed to characterize the coating layers. These YSZ coated and uncoated samples were subjected to wear testing under different spindle speed, loading and working distance. Wear test results were compared with the kinetic friction coefficients and weight loss values. Wear marks on YSZ coated and uncoated samples were investigated by SEM analysis. By coating with plasma spray technique, the wear resistance of Al alloys was increased without changing the friction coefficient. It was found that spindle speed had significant effect over the wear properties than the load applied. By YSZ coating, wear properties were increased 10 times.
The work presents the results of the research and tests of the surface machining of the S355NL and X5CrNi18-10 steels with the concentraded stream of heat with the usage of the GTAW method. The surface layers of the tested steels were remelted with the electric arc using the current of the electric arc 50, 100, 150 and 200A.The machining was done in the atmosphere of argon with the constant speed of the welding head. A microscope examination was performed of the obtained structure and measurements of depth, width and hardness of the received surface layer were performed. Moreover the relation between the current of the electric arc and geometry of the remelted layers with their microhardness was examined.
Fe-40wt% TiB2 nanocomposites were fabricated by mechanical activation and spark-plasma sintering of a powder mixture of iron boride (FeB) and titanium hydride (TiH2). The powder mixture of (FeB, TiH2) was prepared by high-energy ball milling in a planetary ball mill at 700 rpm for 3 h followed by spark-plasma sintering (SPS) at various conditions. Analysis of the change in relative sintered density and densification rate during sintering showed that a self-propagating high-temperature synthesis reaction occurs to form TiB2 from FeB and Ti. A sintered body with relative density higher than 98% was obtained after sintering at 1150°C for 5 and 15 min. The microstructural observation of sintered compacts with the use of FE-SEM and TEM revealed that ultrafine particulates with approximately 5 nm were evenly distributed in an Fe-matrix. A hardness value of 83 HRC was obtained, which is equivalent to that of conventional WC-20 Co systems.
The β-phase Titanium (β-Ti) alloys have been under the spotlight in the recent past for their use as biomedical prosthetic materials owing to their excellent properties such as low elastic modulus, high corrosion resistance and tensile strength. Recently, Niobium (Nb) has gained a lot of attention as a β-phase stabilizing element in Ti alloys to replace Vanadium (V) due to its excellent solubility in Ti, low elastic modulus and biocompatibility. In this work, low cost Ti-20Nb binary alloy has been fabricated via powder metallurgy procedures. The blended powder mixtures of Ti and Nb were sintered at 900°C for 20 mins by the Spark Plasma Sintering (SPS) with an applied uniaxial pressure of 40 MPa. The heating rate was fixed at 50°C/min. The sintered alloy was subject to heat treatments at 1200°C in vacuum condition for various time durations. The characterizations of microstructure obtained during this process were done using FE-SEM, EDS and XRD. By increasing heat treatment time, as understood, the volume of residual Nb particles was decreased resulting in accelerated diffusion of Nb into Ti. Micro hardness of the alloy increased from 340 to 355 HV with the increase in β phase content from 30 to 45%. The resultant alloys had relatively high densities and homogenized microstructures of dispersed lamellar β grains in α matrix.
New graphite tools were designed and produced to fabricate a semi-finished product from which nine cutting inserts were obtained in one spark plasma sintering process. As a result, WC-5Co cemented carbides were spark plasma sintered and the effect of various sintering parameters such as compacting pressure, heating rate and holding time on the main mechanical properties were investigated. It was shown that WC-5Co cemented carbides spark plasma sintered at 1200°C, 80 MPa, 400°C/min, for 5 min are characterized by the best relation of hardness (1861 ±10 HV30) and fracture toughness (9.30 MPa·m1/2). The microstructure of these materials besides the WC ceramic phase and Co binder phase consists of a synthesized Co3W3C complex phase. Comparison with a commercial WC-6Co cutting insert fabricated by conventional powder metallurgy techniques shows that spark plasma sintering is a very effective technique to produce materials characterized by improved mechanical properties.
The microstructure and corrosion properties of spark plasma sintered yttria dispersed and yttria free duplex and ferritic stainless samples were studied. Spark plasma sintering (SPS) was carried out at 1000°C by applying 50 MPa pressure with holding time of 5 minutes. Linear sweep voltammetry (LSV) tests were employed to evaluate pitting corrosion resistance of the samples. Corrosion studies were carried out in 0.5, 1 and 2 M concentration of NaCl and H2SO4 solutions at different quiet time of 2, 4, 6, 8 and 10 seconds. Yttria dispersed stainless steel samples show more resistance to corrosion than yttria free stainless steel samples. Pitting potential decreases with increase in reaction time from 2 to 10 seconds. Similarly, as concentration of NaCl and H2SO4 increases from 0.5 M to 2 M the corrosion resistance decrements due to the availability of more Cl¯ and SO4 2¯ ions at higher concentration.
The effects of carbon content on the austenite stability and strain-induced transformation of nanocrystalline Fe-11% Ni alloys were investigated using X-ray analysis and mechanical tests. The nanocrystalline FeNiC alloy samples were rapidly fabricated using spark plasma sintering because of the extremely short densification time, which not only helped attain the theoretical density value but also prevented grain growth. The increased austenite stability resulted from nanosized crystallites in the sintered alloys. Increasing compressive deformation increased the volume fraction of strain-induced martensite from austenite decomposition. The kinetics of the strain-induced martensite formation were evaluated using an empirical equation considering the austenite stability factor. As the carbon content increased, the austenite stability was enhanced, contributing to not only a higher volume fraction of austenite after sintering, but also to the suppression of its strain-induced martensite transformation.