Normal Size Distribution Research Articles

Effect of Co doping on the magnetic and DC electrical properties of Mn-Zn nanoferrites

In this study, Cobalt-Manganese-Zinc nanoferrites with the formula CoxMn0.5−xZn0.5Fe2O4 with x = 0.0, 0.1, 0.3, and 0.5 prepared by chemical Co-precipitation method. Then the structure and morphology of the synthesized nanoparticles were characterized by X-ray diffraction (XRD) and transmitting electron microscopy (TEM), respectively. The XRD patterns indicated the formation of single-phased cubic structure of spinel ferrite in nanometer size with no minor phase. The TEM image showed the formation of nanoparticles with average size of about 40 nm and normal size distribution. The magnetic measurements of the nanoparticles were done at room temperature using a vibrating sample magnetometer (VSM). Results exhibited a super-paramagnetic like behavior for some of the samples. DC electrical resistivity measurements were carried out by two-probe technique from 25 to 250 °C and showed decreasing of the resistivity with temperature meanwhile passing a transition to form of a peak. The peaks values observed near the Curie temperatures of samples suggest that anomaly behavior can attributed to spin canting associated with the phase transition from para to ferromagnetic state at TC.

Polyacrylic acid-coated iron oxide magnetic nanoparticles: The polymer molecular weight influence

Magnetic nanoparticles (MNPs), in particular, magnetic iron oxide-based nanoparticles were found to be useful as catalysts and as devices for data storage, environmental remediation and several biomedical applications, due to their excellent properties, such as biocompatibility and high magnetic moment. Polyacrylic acid (PAA) is a weak polyelectrolyte that can be used to stabilize the MNPs. To the best of our knowledge, the influence of PAA molecular weight and PAA concentration over the magnetic and structural properties of iron oxide nanoparticles has not been previously reported. The aim of this paper is to describe the differences evidenced in the properties of different magnetic materials by using PAA for iron oxides stabilization by one-pot coprecipitation synthesis. Iron oxide-based magnetic nanoparticles stabilized by polyacrylic acid (PAA) polymers were efficiently prepared and exhaustively characterized. The influence on the employment of two different low PAA molecular weights, Mw 1800 g/mol and 5000 g/mol, in three different iron salts: PAA ratios was analyzed. In summary, the main results showed that: for a certain PAA reactor feed higher oligomer quantities are present in MNPs as higher is the involved molecular weight of the polymeric chain; when molecular weight raises the contribution of loops and tails also does it, allowing having higher polymer contents. For both PAA’s Mw employed as the adsorbed PAA increases particles hydrodynamic diameters decreases, and their distribution becomes narrower; the PAA adsorbs onto iron oxides by chemisorption (the most probable interaction is the bidentate bridging). For the studied cases z potential values depend much more on the PAA’s quantity adsorbed onto the iron oxides than on the PAA’s Mw. MNPs are superparamagnetic and choosing the right shape of particle distribution is not central for getting estimates of the magnetization saturation, the average particle diameter and its standard deviation, while better fits are found with Normal and Log-Normal particle size distributions.

The control of abnormal grain growth in low-voltage SnO2 varistors by microseed addition

In this research, the addition effects of three different quantities of micron-sized seeds (microseeds) to a SnO2 varistor prepared from nanomaterials on the microstructure and electrical properties were studied. Moreover, surge-withstanding capability of low-voltage SnO2 varistors was investigated. The X-ray diffraction pattern disclosed a single phase SnO2 for microseed grains. The morphological features of samples were characterized using scanning electron microscopy. The abnormal distribution of grain size with elongated grains of SnO2 in fine grains matrix was observed in sintered samples without microseeds. The low content of microseed addition (0.3wt%) had not controlled abnormal grain growth, however, it increased mean grain size to 37µm. Although the high content of microseeds (7.5wt%) stopped abnormal grain growth, it had a negative effect on relative density and mean grain size. The normal grain size distribution with maximum mean grain size (45µm) was obtained in samples containing 1.5wt% microseeds. These samples showed the lowest breakdown field (240V/cm) and the highest surge-withstanding capability (1.5kA/cm2). Furthermore, the standard deviation of the electrical parameters of these samples was improved due to normal grain-size distribution.

INVESTIGATION OF THE FORMATION PROCESS OF HAZARDOUS AND HARMFUL PRODUCTION FACTORS WHEN CUTTING A STONE FOR CONSTRUCTION WORKS

Stone cutting for construction work is carried out by disk diamond wheels the rotation speed of which, and, consequently, the cutting speed is 35-50 m/s. In view of the high intensity of the cutting process and intensive microchip formation, the process of stone cutting is accompanied by considerable dust formation, which can be both harmful and dangerous in the work. The greatest danger is represented by dust particles, which dimensions are 5 μm or less. These particles have the greatest pathogenic effect on the respiratory system of the human body. In addition, the settling time of these particles is measured in hours. Thus, even after the termination of work, the risk of dust exposure to the human body remains. Insignificant time of inhalation of these particles can lead a person to disability and death. Insurance payments in case of disability and compensation in case of death of an employee for these reasons all over the world amount to multibillion sums, which, even for this reason, makes the issue of combating this phenomenon very urgent. In this paper, the process of generation of hazardous and harmful production factors arising as a result of dust formation during the stone cutting, the regularity of dust distribution by fractions, quantitative indicators of the process for improving equipment, as well as individual and collective means of personnel protection are determined. The shape and dimensions of the cutting grains of the cutting discs are studied in the paper, depending on the grain material and the grain size of the cutting disc. Thecurvature radii of the cutting grains depend on the grain material and the grain size of the cutting disc. The actual number of grains participating in cutting and, consequently, in dust formation is shown. The dimensions of the dust particles depend on the graininess of the cutting discs and the processing regimes. Using the law of normal particle size distribution, the percentage of dust particles is determined depending on processing conditions. The rate of particle settling depends on their size and mass. The formation of a dust-air mixture and its probable concentration and chemical composition of dust depend on the chemical composition of the cutting materials. Regularities are obtained, which can give an opportunity to improve the individual and collective protection of workers from this harmful production factor.

Influence of oxygen availability on the metabolism and morphology of Yarrowia lipolytica: insights into the impact of glucose levels on dimorphism.

Dynamic behavior of Yarrowia lipolytica W29 strain under conditions of fluctuating, low, and limited oxygen supply was characterized in batch and glucose-limited chemostat cultures. In batch cultures, transient oscillations between oxygen-rich and -deprived environments induced a slight citric acid accumulation (lower than 29mgL-1). By contrast, no citric acid was detected in continuous fermentations for all stress conditions: full anoxia (zero pO2 value, 100% N2), limited (zero pO2 value, 75% of cell needs), and low (pO2 close to 2%) dissolved oxygen (DO) levels. The macroscopic behavior (kinetic parameters, yields, viability) of Y. lipolytica was not significantly affected by the exposure to DO fluctuations under both modes of culture. Nevertheless, conditions of oxygen limitation resulted in the destabilization of the glucose-limited growth during the continuous cultivations. Morphological responses of Y. lipolytica to DO oscillations were different between batch and chemostat runs. Indeed, a yeast-to-mycelium transition was induced and progressively intensified during the batch fermentations (filamentous subpopulation reaching 74% (v/v)). While, in chemostat bioreactors, the culture consisted mainly of yeast-like cells (mean diameter not exceeding 5.7μm) with a normal size distribution. During the continuous cultures, growth at low DO concentration did not induce any changes in Y. lipolytica morphology. Dimorphism (up to 80.5% (v/v) of filaments) was only detected under conditions of oxygen limitation in the presence of a residual glucose excess (more than 0.75gL-1). These data suggest an impact of glucose levels on the signaling pathways regulating dimorphic responses in Y. lipolytica.

Superporous pellicular κ-Carrageenan–Nickel composite beads; morphological, physical and hydrodynamics evaluation for expanded bed adsorption application

Expanded bed adsorption (EBA) is an authentic separation method for the purification of bioproducts. Novel pellicular κ-Carrageenan–Nickel (KC–Ni) and superporous κ-Carrageenan–Nickel (SP-KC–Ni) adsorbent beads have been fabricated and evaluated using water in oil emulsification and granule leaching methods. The morphology, structure and porosity of the fabricated composite matrices were investigated. A proper spherical shape and porous structure with nano sized pores in the range of about 50–190nm has been detected for the KC–Ni beads and upper than 50% of the SP-KC–Ni superpores was at the range of 600–5400nm. The results from the image processing indicated that the KC–Ni particles had normal size distribution with the range of 60–320μm. As a result of physical properties measurements, a proper wet density at the range of 1.43–2.42g/mL was resulted for the adsorbent beads. The porosity and effective porosity of egg albumin (EA) for SP-KC–Ni3 was measured about 98.89% and 73.87% respectively. Residence time distribution (RTD) measurements were applied to investigate hydrodynamic characteristics of KC–Ni and SP-KC–Ni matrices in expanded bed. The results showed that proper physical and hydrodynamic characteristics of KC–Ni and SP-KC–Ni matrices confirm the potential of use in high speed expanded bed chromatography.

Influence of Bi3+content on photoluminescence of InNbO4:Eu3+,Bi3+for white light-emitting diodes

AbstractA series of red-emitting phosphors InNbO4:Eu3+,Bi3+was prepared by a high temperature solid-state reaction. The structure, size distribution and luminescence properties of the phosphors were respectively characterized by X-ray diffraction (XRD), laser particle size and molecular fluorescence spectrometer. The XRD results indicate that the phase-pure samples have been obtained and the crystal structure of the host has not changed under the Eu3+and Bi3+co-doping. The test of size distribution shows that the phosphor has a normal size distribution. The excitation spectra illustrate that the dominant sharp peaks are located at 394 nm (7F0→5L6) and 466 nm (7F0→5D2). Meanwhile, the emission spectra reveal that the phosphors excited by the wavelength of 394 nm or 466 nm have an intense red-emission line at 612 nm owing to the5D0→7F2transition of Eu3+. Bi3+doping has not changed the peak positions except the photoluminescence intensity. The emission intensity is related to Bi3+concentration, and it is up to the maximum when the Bi3+-doping concentration is 4 mol%. Due to good photoluminescence properties of the phosphor, the InNbO4:0.04Eu3+,0.04Bi3+may be used as a red component for white light-emitting diodes.

Non-isothermal crystallization kinetics assessment of poly(lactic acid)/graphene nanocomposites

In this work, the effects of the presence and modification of graphene nanoplatelets (GNps) on the crystallization of the poly(lactic acid) (PLA) were studied. Functionalization of GNps was accomplished by acid treatment. Nanocomposite samples were prepared by solution method containing pristine and functionalized graphene. In contrast to pristine PLA, crystallization of the samples contains nano filler initiates at higher rates that showed the role of heterogeneous nucleating effects of these particles in crystallization of the PLA. Then, the effect of nano filler functionalization was comprised. Initial slope of the crystallization (Si) and full width at the half height maximum of crystallization peak are indicative of nucleation rate and spherulite size distribution, respectively; which upon the addition of the functionalized graphene nanoplatelets (FGNps), Si increased and spherulites gained normal size distribution. Non-isothermal and crystallization kinetics of the samples were studied using differential scanning calorimetry at heating rates of 2, 4, 6 and 10 °C/min. Performed techniques such as furrier transform infrared, dynamic-mechanical thermal analysis and visual observation of sediments confirmed the successful modification of the graphene platelets. Also, non-isothermal analysis pinpointed the fact that crystallization temperature (Tc) of the nanocomposites has increased by 11–21 °C, compared to the neat PLA. Upon verification of Avrami’s theory, it was conducted that dominant mechanism of nucleation of the nanocomposite samples was 2D circular diffusion; wherein, Avrami’s exponent (n) was determined as 2. Moreover, it was deduced from Avrami’s equation that “n” have no discernible changes in nanocomposites containing GNps or FGNps. Electrical devices and shape memories can be the main application of these nanocomposites.

Reactive sintering and microstructure development of tungsten carbide-AISI 304 stainless steel cemented carbides

Sintering of WC-stainless steel (SS) composites within a typical binder range from 6 up to 15 wt% SS was investigated through constant heating rate dilatometry, in vacuum conditions, complemented by differential thermal analysis and by the study of the high temperature wetting behavior of SS on WC. The densification starts ∼900 °C with a typical densification curve for all compositions, where three distinct regions are discernible: the first one with a slow densification rate, followed by a second region where a sharp increase in the densification rate up to a maximum value dependent on the binder amount is observed and, finally, a third one with a slowdown of the densification rate until the end of the thermal cycle. The attained final density at 1450 °C is dependent on the binder amount, increasing proportionally to its initial content. The final microstructure presents a normal grain size distribution and appreciable amounts of eta-phase, besides the major WC phase and residual iron rich phase. The reactive densification behavior and the role of the liquid phase are interpreted accordingly with structural and kinetic data.

English

The study estimated the amount of energy required for grinding palm kernel shell and groundnut shell using laboratory hammer mill and also characterize their ground physical properties. The material type and hammer mill screen aperture significantly influenced the energy requirement for grinding the two materials. For a given screen aperture size, groundnut shell consumed more energy than palm kernel shell. The energy relationship with hammer mill screen exhibited a second order polynomial form. Within the same hammer mill screen aperture range, the energy requirements for palm kernel and groundnut shell were considerably lower than most grass based biomass or straw. The bulk and particle density of groundnut shell decreased with increased geometric mean diameter while that of palm kernel shell increased with geometric mean. The two materials ground exhibited a lognormal particle size distribution at hammer mill screen of 5 and 3 mm, while 0.8 mm screen aperture exhibited a normal size distribution. The total particle and surface area estimate in a charge was sensitive to the screen sizes. Energy equations: Ratzinger’s, Kicks and Bond’s were used to investigate the results of the biomass comminution with Ratzinger’s equations showing a higher R2 value for palm kernel shell while Kick’s equation showed a higher R2 value for groundnut shell. Key words: Milling, energy consumption, groundnut shell, palm kernel shell, grinding.

Surface Plasmon Resonance Imaging Microscopy of Liposomes and Liposome-Encapsulated Gold Nanoparticles

Liposomes are small vesicles that can be used in various targeting applications as carrier vehicles. In this paper, we show that real-time surface plasmon resonance imaging microscopy (SPRI microscopy) can be used to detect diffraction patterns of these singular vesicles in water phase at room temperature and without any additives. The diffraction pattern intensities, related to the particle size, are shown to follow the log-normal distribution in a cumulative distribution function (CDF) that is very well in accordance with the normal size distribution of liposomes prepared with the extrusion method. In addition, this distribution is further analyzed to determine the number of gold nanoparticle (GNP) encapsulated liposomes in a set of liposomal adsorption events. Thus, we obtain the encapsulation efficiency and present a method to study the intrinsic properties of liposomes and other soft nanomaterials.

Carboxymethylated lignins with low surface tension toward low viscosity and highly stable emulsions of crude bitumen and refined oils

Kraft and organosolv lignins were subjected to carboxymethylation to produce fractions that were soluble in water, displayed a minimum surface tension as low as 34mN/m (25°C) and a critical aggregation concentration of ∼1.5wt%. The carboxymethylated lignins (CML), which were characterized in terms of their degree of substitution (31P NMR), elemental composition, and molecular weight (GPC), were found suitable in the formulation of emulsions with bitumens of ultra-high viscosity, such as those from the Canadian oil sands. Remarkably, the interfacial features of the CML enabled fuel emulsions that were synthesized in a very broad range of internal phase content (30–70%). Cryo-replica transmission electron microscopy, which was used here the first time to assess the morphology of the lignin-based emulsions, revealed the droplets of the emulsion stabilized with the modified lignin. The observed drop size (diameters<2μm) was confirmed by light scattering, which revealed a normal size distribution. Such characteristics led to stable emulsified systems that are amenable for a wide range of applications. Emulsification with CML afforded bitumen emulsions with very high colloidal stability (no change was noted for over one month) and with a strong shear thinning behavior. Both features indicate excellent prospects for storage, transport and spraying, which are relevant in operations for power generation, which also take advantage of the high heating value of the emulsion components. The ability of CML to stabilize emulsions and to contribute in their combustion was tested with light fuels (kerosene, diesel, and jet fuel) after formulation of high internal phase systems (70% oil) that enabled operation of a fuel engine. A significant finding is that under certain conditions and compared to the respective pure fuel, combustion of the O/W emulsions stabilized by CML presented lower NOx and CO emissions and maintained a relatively high combustion efficiency. The results highlight the possibilities in high volume application for lignin biomacromolecules.

Numerical study of different particle size distribution for modeling of solid-liquid extraction in randomly packed beds

Abstract In this paper, the Montan wax extraction as an industrial example for a solid-liquid extraction process is modeled and simulated using 2D and 3D CFD models. Carrying out various simulation studies, the applied particle size distribution of the solid phase is studied concerning its impact on the extraction kinetics and the extraction rates. In the modeling, the solid phase, which is a wax-containing brown coal, is described by establishing 2D models of random packing in cylindrically packed beds. In the study, the applicability of a 2D CFD model is validated against simulation result of a 3D model of a random packing. The random packing of the 2D model considers the same mean particle size with a uniform distribution and a normal distribution, respectively. The simulation result indicates that the extraction rate is greatly increased using the brown coal of a normal particle size distribution, which is a good match with experimental results.

Arrange and Average Algorithm for Microphysical Retrievals with A “3β+3α” Lidar Configuration

We present the results of a comparison study in which a simple, automated, and unsupervised algorithm, which we call the arrange and average algorithm, was used to infer microphysical parameters (complex refractive index (CRI), effective radius, total number, surface area, and volume concentrations) of atmospheric aerosol particles. The algorithm normally uses backscatter coefficients ( β ) at 355, 532, and 1064 nm and extinction coefficients ( α ) at 355 and 532 nm as input information. We compared the performance of the algorithm for the existing “3 β +α” and potential “3 β +3α” configurations of a multiwavelength aerosol Raman lidar or highspectral-resolution lidar (HSRL). The “3 β +3α” configuration uses an extra extinction coefficient at 1064 nm. Testing of the algorithm is based on synthetic optical data that are computed from prescribed CRIs and monomodal logarithmically normal particle size distributions that represent spherical, primarily fine mode aerosols. We investigated the degree to which the microphysical results retrieved by this algorithm benefits from the increased number of input extinction coefficients.

Potential source identification for aerosol concentrations over a site in Northwestern India

The collocated measurements of aerosols size distribution (ASD) and aerosol optical thickness (AOT) are analyzed simultaneously using Grimm aerosol spectrometer and MICROTOP II Sunphotometer over Jaipur, capital of Rajasthan in India. The contrast temperature characteristics during winter and summer seasons of year 2011 are investigated in the present study. The total aerosol number concentration (TANC, 0.3–20μm) during winter season was observed higher than in summer time and it was dominated by fine aerosol number concentration (FANC<2μm). Particles smaller than 0.8μm (at aerodynamic size) constitute ~99% of all particles in winter and ~90% of particles in summer season. However, particles greater than 2μm contribute ~3% and ~0.2% in summer and winter seasons respectively. The aerosols optical thickness shows nearly similar AOT values during summer and winter but corresponding low Angstrom Exponent (AE) values during summer than winter, respectively. In this work, Potential Source Contribution Function (PSCF) analysis is applied to identify locations of sources that influenced concentrations of aerosols over study area in two different seasons. PSCF analysis shows that the dust particles from Thar Desert contribute significantly to the coarse aerosol number concentration (CANC). Higher values of the PSCF in north from Jaipur showed the industrial areas in northern India to be the likely sources of fine particles. The variation in size distribution of aerosols during two seasons is clearly reflected in the log normal size distribution curves. The log normal size distribution curves reveals that the particle size less than 0.8μm is the key contributor in winter for higher ANC.

Microprocessor Complex Subunit DiGeorge Syndrome Critical Region Gene 8 (Dgcr8) Is Required for Schwann Cell Myelination and Myelin Maintenance

We investigated the role of a key component of the Microprocessor complex, DGCR8, in the regulation of myelin formation and maintenance. We found that conditionally ablating Dgcr8 in Schwann cells (SCs) during development results in an arrest of SC differentiation. Dgcr8 conditional knock-out (cKO) SCs fail to form 1:1 relationships with axons or, having achieved this, fail to form myelin sheaths. The expression of genes normally found in immature SCs, such as sex-determining region Y-box 2 (Sox2), is increased in Dgcr8 cKO SCs, whereas the expression of myelin-related genes, including the master regulatory transcription factor early growth response 2 (Egr2), is decreased. Additionally, expression of a novel gene expression program involving sonic hedgehog (Shh), activated de novo in injured nerves, is elevated in Dgcr8 cKOs but not in Egr2 null mice, a model of SC differentiation arrest, suggesting that the injury-related gene expression program in Dgcr8 cKOs cannot be attributed to differentiation arrest. Inducible ablation of Dgcr8 in adult SCs results in gene expression changes similar to those found in cKOs, including an increase in the expression of Sox2 and Shh. Analyses of these nerves mainly reveal normal myelin thickness and axon size distribution but some dedifferentiated SCs and increased macrophage infiltration. Together our data suggest that Dgcr8 is responsible for modulation of gene expression programs underlying myelin formation and maintenance as well as suppression of an injury-related gene expression program.

Autophagy is induced upon platelet activation and is essential for hemostasis and thrombosis

AbstractAutophagy is important for maintaining cellular homeostasis, and thus its deficiency is implicated in a broad spectrum of human diseases. Its role in platelet function has only recently been examined. Our biochemical and imaging studies demonstrate that the core autophagy machinery exists in platelets, and that autophagy is constitutively active in resting platelets. Moreover, autophagy is induced upon platelet activation, as indicated by agonist-induced loss of the autophagy marker LC3II. Additional experiments, using inhibitors of platelet activation, proteases, and lysosomal acidification, as well as platelets from knockout mouse strains, show that agonist-induced LC3II loss is a consequence of platelet signaling cascades and requires proteases, acidic compartments, and membrane fusion. To assess the physiological role of platelet autophagy, we generated a mouse strain with a megakaryocyte- and platelet-specific deletion of Atg7, an enzyme required for LC3II production. Ex vivo analysis of platelets from these mice shows modest defects in aggregation and granule cargo packaging. Although these mice have normal platelet numbers and size distributions, they exhibit a robust bleeding diathesis in the tail-bleeding assay and a prolonged occlusion time in the FeCl3-induced carotid injury model. Our results demonstrate that autophagy occurs in platelets and is important for hemostasis and thrombosis.

Cross-linked κ-carrageenan polymer/zinc nanoporous composite matrix for expanded bed application: Fabrication and hydrodynamic characterization

Expanded bed adsorption (EBA) is a reliable separation technique for the purification of bioproducts from complex feedstocks. The specifically designed adsorbent is necessary to form a stable expanded bed. In the present work, a novel custom-designed composite matrix has been prepared through the method of water-in-oil emulsification. In order to develop an adsorbent with desirable qualities and reduce the costs, κ-carrageenan and zinc powder were used as the polymeric skeleton and the densifier, respectively. The prepared composite matrix was named as KC-Zn. Optical microscope (OM) and scanning electron microscope (SEM) were applied to characterize the morphology and structure of prepared composite matrix. These analyses approved good spherical shape and porous structure with nano-scale pores in the range of about 60–180nm. The results from the particle size analyzer (PSA) revealed that all the KC-Zn beads followed logarithmic normal size distribution with the range of 50–350μm and average diameter of 160–230μm, respectively. Main physical properties of KC-Zn matrices were measured as a function of zinc powder ratio to κ-carrageenan slurry, which showed an appropriate wet density in the range of 1.39–2.27g/ml, water content of 72.67–36.41% and porosity of 98.07–80.24%, respectively. The effects of matrix density and liquid phase viscosity on hydrodynamic behavior of prepared matrix have been investigated by residence time distribution (RTD) experiments in an expanded bed. The results indicated that in a constant liquid velocity as the matrix density was increased, the expansion factor of bed decreased and the axial mixing coefficient increased. Moreover, an enhancement in the fluid viscosity led to an increase in the bed expansion and a decrease in the stability of expanded bed. Therefore using a matrix with higher density seems necessary to face viscous feedstocks. All the results demonstrated that proper physical properties and hydrodynamic characteristics of KC-Zn matrix confirm good potential for possible use in high flow rate expanded bed operations.

Study of Structural Features of Porous TINI-Based Materials Produced by SHS and Sintering

Structural properties of porous TiNi-based materials produced by SHS method and sintering have been investigated. The material having different pore wall surface topography, porosity and pore size distribution was shown to be produced depending on the powder metallurgy method for porous TiNi-based alloy. All the materials having porosity of 55-70%, mean pore size 90-150 μm, as well as normal pore size distribution are most preferable. Ultimate strength and breaking point were determined to depend on porosity, pore size distribution, pore intersections and phase chemical composition of the material. Strength properties of the sintered alloy are twice as much compared to the SHS-produced ones due to homogeneity of its macrostructure, low chemical heterogeneity and TiNi3 precipitations strengthening the TiNi matrix.

ELISPOTs Produced by CD8 and CD4 Cells Follow Log Normal Size Distribution Permitting Objective Counting.

Each positive well in ELISPOT assays contains spots of variable sizes that can range from tens of micrometers up to a millimeter in diameter. Therefore, when it comes to counting these spots the decision on setting the lower and the upper spot size thresholds to discriminate between non-specific background noise, spots produced by individual T cells, and spots formed by T cell clusters is critical. If the spot sizes follow a known statistical distribution, precise predictions on minimal and maximal spot sizes, belonging to a given T cell population, can be made. We studied the size distributional properties of IFN-γ, IL-2, IL-4, IL-5 and IL-17 spots elicited in ELISPOT assays with PBMC from 172 healthy donors, upon stimulation with 32 individual viral peptides representing defined HLA Class I-restricted epitopes for CD8 cells, and with protein antigens of CMV and EBV activating CD4 cells. A total of 334 CD8 and 80 CD4 positive T cell responses were analyzed. In 99.7% of the test cases, spot size distributions followed Log Normal function. These data formally demonstrate that it is possible to establish objective, statistically validated parameters for counting T cell ELISPOTs.