Loggerhead sea turtle Caretta caretta plasma biochemistry and proteome profile modulation during recovery.

The aim of the study was to monitor and analyse injured and diseased loggerhead sea turtles (Caretta caretta) plasma proteome profiles and biochemistry parameters during their recovery period in rescue centre within different age and recovery period groups, and determine the potential biomarkers that can be used in diagnostics. The plasma biochemical parameters of total protein and glucose content, accompanied by aspartate aminotransferase (AST) and N-acetyl-cystein-activated creatinine kinase (CK-NAC) are highlighted as valuable and potential biomarkers of turtle's health status and condition. Using high throughput tandem mass tag (TMT)-based proteomic approach we identified 913 plasma proteins, 12 of which shown to be modulated in loggerheads age groups, and identified as a part of (i) platelet degranulation, (ii) neutrophil degranulation, and (iii) innate immune system pathways. The neurofascin (NFASC) is shown to be differentially abundant among all the age groups, and alpha-1-acid glycoprotein 2-like (ORM2) and alpha-1-antitrypsin-like (SERPINA1) proteins were recognized as members of all three above mentioned REACTOME pathways. Furthermore, 29 of plasma proteins were significantly differentially abundant in loggerheads age and recovery period groups. Out of 15 recognized pathways, those proteins were mostly included in three specific REACTOME pathways: (i) post-translational phosphorylation, (ii) regulation of Insulin-like Growth Factor (IGF) transport and uptake by Insulin-like Growth Factor Binding Proteins (IGFBPs), and (iii) platelet degranulation. The alpha-fetoprotein (AFP) was the only protein which showed statistically significant up-regulation patterns in all loggerhead age groups before release from the rescue centre, and the complement component 3 (C3) protein was the only protein modulated in all recovery period groups. Furthermore, C3 protein takes part in 9; and followed up with apolipoprotein A-I (APOA1) in 7; complement component 4 (C4), complement component 5 (C5) and kininogen-1 (KNG1) in 6 REACTOME pathways. Thereby, those proteins are highlighted and recommended as potential biomarkers of turtle's health status. Data are available via ProteomeXchange with identifier PXD029569. Finally, based on our results, we believe that comprehensive omics approach and routine plasma biochemical analysis, accompanied by proteins of acute phase, acid-base status and immune-response indicator analysis may significantly and reliably improve assessment of captive loggerheads rehabilitation and medication. SIGNIFICANCE: Monitoring and comparison of loggerhead sea turtles (C. caretta) blood plasma biochemistry parameters and plasma proteome profiles in relation to the age, and recovery period pointed out significantly differentially abundant proteins, along with certain biochemical parameter contents as potential biomarkers of turtle's fitness, health status and physiology.

Random sequential adsorption of lattice animals on a three-dimensional cubic lattice.

The properties of the random sequential adsorption of objects of various shapes on simple three-dimensional (3D) cubic lattice are studied numerically by means of Monte Carlo simulations. Depositing objects are "lattice animals," made of a certain number of nearest-neighbor sites on a lattice. The aim of this work is to investigate the impact of the geometrical properties of the shapes on the jamming density θ_{J} and on the temporal evolution of the coverage fraction θ(t). We analyzed all lattice animals of size n=1, 2, 3, 4, and 5. A significant number of objects of size n⩾6 were also used to confirm our findings. Approach of the coverage θ(t) to the jamming limit θ_{J} is found to be exponential, θ_{J}-θ(t)∼exp(-t/σ), for all lattice animals. It was shown that the relaxation time σ increases with the number of different orientations m that lattice animals can take when placed on a cubic lattice. Orientations of the lattice animal deposited in two randomly chosen places on the lattice are different if one of them cannot be translated into the other. Our simulations performed for large collections of 3D objects confirmed that σ≅m∈{1,3,4,6,8,12,24}. The presented results suggest that there is no correlation between the number of possible orientations m of the object and the corresponding values of the jamming density θ_{J}. It was found that for sufficiently large objects, changing of the shape has considerably more influence on the jamming density than increasing of the object size.

Particle morphology effects in random sequential adsorption.

The properties of the random sequential adsorption of objects of various shapes on a two-dimensional triangular lattice are studied numerically by means of Monte Carlo simulations. The depositing objects are formed by self-avoiding lattice steps, whereby the size of the objects is gradually increased by wrapping the walks in several different ways. The aim of this work is to investigate the impact of the geometrical properties of the shapes on the jamming density θ_{J} and on the temporal evolution of the coverage fraction θ(t). Our results suggest that the order of symmetry axis of a shape exerts a decisive influence on adsorption kinetics near the jamming limit θ_{J}. The decay of probability for the insertion of a new particle onto a lattice is described in a broad range of the coverage θ by the product between the linear and the stretched exponential function for all examined objects. The corresponding fitting parameters are discussed within the context of the shape descriptors, such as rotational symmetry and the shape factor (parameter of nonsphericity) of the objects. Predictions following from our calculations suggest that the proposed fitting function for the insertion probability is consistent with the exponential approach of the coverage fraction θ(t) to the jamming limit θ_{J}.

Assessing silver nanoparticles behaviour in artificial seawater by mean of AF4 and spICP-MS.

The use of nanotechnology-based products is constantly increasing and there are concerns about the fate and effect on the aquatic environment of antimicrobial products such as silver nanoparticles. By combining different characterization techniques (asymmetric flow field-flow fractionation, single particle ICP-MS, UV-Vis) we show that it is possible to assess in detail the agglomeration process of silver nanoparticles in artificial seawater. In particular we show that the presence of alginate or humic acid differentially affects the kinetic of the agglomeration process. This study provides an experimental methodology for the in-depth analysis of the fate and behaviour of silver nanoparticles in the aquatic environment.

Structural characterization of submerged granular packings.

We consider the impact of the effective gravitational acceleration on microstructural properties of granular packings through experimental studies of spherical granular materials saturated within fluids of varying density. We characterize the local organization of spheres in terms of contact connectivity, distribution of the Delaunay free volumes, and the shape factor (parameter of nonsphericity) of the Voronoï polygons. The shape factor gives a clear physical picture of the competition between less and more ordered domains of particles in experimentally obtained packings. As the effective gravity increases, the probability distribution of the shape factor becomes narrower and more localized around the lowest values of the shape factor corresponding to regular hexagon. It is found that curves of the pore distributions are asymmetric with a long tail on the right-hand side, which progressively reduces while the effective gravity gets stronger for lower densities of interstitial fluid. We show that the distribution of local areas (Voronoï cells) broadens with decreasing value of the effective gravity due to the formation of lose structures such as large pores and chainlike structures (arches or bridges). Our results should be particularly helpful in testing the newly developed simulation techniques involving liquid-related forces associated with immersed granular particles.

Optimization of the monolayer growth in adsorption-desorption processes.

Kinetics of the deposition process of dimers in the presence of desorption is studied by Monte Carlo method on a one-dimensional lattice. The aim of this work is to investigate how do various temporal dependencies of the desorption rate hasten or slow down the deposition process. The growth of the coverage θ(t) above the jamming limit to its steady-state value θ(∞) is analyzed when the desorption probability P(des) decreases both stepwise and linearly (continuously) over a certain time domain. We report a numerical evidence that the time needed for a system to reach the given coverage θ can be significantly reduced if P(des) decreases in time. Finally, a self-consistent optimization procedure, when the probability P(des) depends on the current coverage density θ(t), is formulated and tested. The present model reproduces qualitatively the densification kinetics and the memory effects of vibrated granular materials. Our results suggest that the process of vibratory compaction of granular materials can be optimized by using a time dependent intensity of external excitations.

Percolation in random sequential adsorption of extended objects on a triangular lattice.

The percolation aspect of random sequential adsorption of extended objects on a triangular lattice is studied by means of Monte Carlo simulations. The depositing objects are formed by self-avoiding lattice steps on the lattice. Jamming coverage θ{jam}, percolation threshold θ{p}, and their ratio θ{p}/θ{jam} are determined for objects of various shapes and sizes. We find that the percolation threshold θ{p} may decrease or increase with the object size, depending on the local geometry of the objects. We demonstrate that for various objects of the same length, the threshold θ{p} of more compact shapes exceeds the θ{p} of elongated ones. In addition, we study polydisperse mixtures in which the size of line segments making up the mixture gradually increases with the number of components. It is found that the percolation threshold decreases, while the jamming coverage increases, with the number of components in the mixture.

Simulation study of anisotropic random sequential adsorption of extended objects on a triangular lattice.

The properties of the anisotropic random sequential adsorption (RSA) of objects of various shapes on a two-dimensional triangular lattice are studied numerically by means of Monte Carlo simulations. The depositing objects are formed by self-avoiding lattice steps, whereby the first step determines the orientation of the object. Anisotropy is introduced by positing unequal probabilities for orientation of depositing objects along different directions of the lattice. This probability is equal p or (1-p)/2, depending on whether the randomly chosen orientation is horizontal or not, respectively. Approach of the coverage θ(t) to the jamming limit θ(jam) is found to be exponential θ(jam)-θ(t)is proportional to exp(-t/σ), for all probabilities p. It was shown that the relaxation time σ increases with the degree of anisotropy in the case of elongated and asymmetrical shapes. However, for rounded and symmetrical shapes, values of σ and θ(jam) are not affected by the presence of anisotropy. We finally analyze the properties of the anisotropic RSA of polydisperse mixtures of k-mers. Strong dependencies of the parameter σ and the jamming coverage θ(jam) on the degree of anisotropy are obtained. It is found that anisotropic constraints lead to the increased contribution of the longer k-mers in the total coverage fraction of the mixture.

Relaxation properties in a diffusive model of k-mers with constrained movements on a triangular lattice.

We study the relaxation process in a two-dimensional lattice gas model, based on the concept of geometrical frustration. In this model the particles are k-mers that can both randomly translate and rotate on the planar triangular lattice. In the absence of rotation, the diffusion of hard-core particles in crossed single-file systems is investigated. We monitor, for different densities, several quantities: mean-square displacement, the self-part of the van Hove correlation function, and the self-intermediate scattering function. We observe a considerable slowing of diffusion on a long-time scale when suppressing the rotational motion of k-mers; our system is subdiffusive at intermediate times between the initial transient and the long-time diffusive regime. We show that the self-part of the van Hove correlation function exhibits, as a function of particle displacement, a stretched exponential decay at intermediate times. The self-intermediate scattering function (SISF), displaying slower than exponential relaxation, suggests the existence of heterogeneous dynamics. For each value of density, the SISF is well described by the Kohlrausch-Williams-Watts law; the characteristic timescale τ(q(n)) is found to decrease with the wave vector q(n) according to a simple power law. Furthermore, the slowing of the dynamics with density ρ(0) is consistent with the scaling law 1/τ(q(n);ρ(0))∝(ρ(c)-ρ(0))(Ï°), with the same exponent Ï°=3.34±0.12 for all wave vectors q(n). The density ρ(c) is approximately equal to the closest packing limit, θ(CPL)≲1, for dimers on the two-dimensional triangular lattice. The self-diffusion coefficient D(s) scales with the same power-law exponent and critical density.

Induction of apoptosis in mussel Mytilus galloprovincialis gills by model cytotoxic agents.

Apoptosis signaling pathway was investigated in the marine mussel Mytilus galloprovincialis exposed to various stressors. Analyses were performed in mussels exposed to two major pollutants of the aquatic environment: tributyltin and the water soluble fraction of diesel oil, for 1 h and animals were then maintained in sea water for a recovery period of 6 and 24 h. Apoptosis was evaluated at several levels of the cell signaling cascade by measuring Bcl-xS expression, caspase-3 activity and DNA damage (Fast micromethod(®) and TUNEL techniques). H(2)O(2) was used as a control of apoptosis induction for validation of the assays. Results showed an induction of Bcl-xS expression, a protein implicated in apoptosis, after 1 h exposure to all concentrations of chemicals. Moreover, in the same manner, apoptotic DNA damage was induced with all chemicals tested. Besides, caspase 3 activity was detected after 1 h exposure to low doses of TBT and diesel oil while the high concentrations induced this protein after 6 h. The achieved data were also correlated with our previous study, demonstrating an induction of the mitogen-activated protein kinase (MAPK) activity in the mussel M. galloprovincialis exposed to the same conditions. In conclusion, this study was one of the first characterizing the MAP kinase cell signaling pathway leading to apoptosis in the mussel M. galloprovincialis exposed to chemicals. It showed for the first time that the Bcl-xS protein was present in these mussels as in other species and played a role in apoptosis mediation. Moreover, the main originality of this work was that it showed that two apoptotic pathways might be present in the mussel: a caspase 3-dependent and a caspase 3-independent pathways.

Upward penetration of grains through a granular medium.

We study experimentally the creeping penetration of guest (percolating) grains through densely packed granular media in two dimensions. The evolution of the system of the guest grains during the penetration is studied by image analysis. To quantify the changes in the internal structure of the packing, we use Voronoï tessellation and a certain shape factor which is a clear indicator of the presence of different underlying substructures (domains). We first consider the impact of the effective gravitational acceleration on upward penetration of grains. It is found that the higher effective gravity increases the resistance to upward penetration and enhances structural organization in the system of the percolating grains. We also focus our attention on the dependence of the structural rearrangements of percolating grains on some parameters like polydispersity and the initial packing fraction of the host granular system. It is found that the anisotropy of penetration is larger in the monodisperse case than in the bidisperse one, for the same value of the packing fraction of the host medium. Compaction of initial host granular packing also increases anisotropy of penetration of guest grains. When a binary mixture of large and small guest grains is penetrated into the host granular medium, we observe size segregation patterns.

Thiopurine S-methyltransferase pharmacogenetics: genotype to phenotype correlation in the Slovenian population.

The toxicity of thiopurine drugs has been correlated to the activity of thiopurine S-methyltransferase (TPMT), whose interindividual variation is a consequence of genetic polymorphisms. We have herein investigated the relevance of some genetic markers for the prediction of thiopurine-related toxicities and to determine the genotype to phenotype correlation in the Slovenian population. The most prevalent mutant allele in the Slovenian population is TPMT*3A (4.1%), followed by TPMT*3C (0.5) and TPMT*3B (0.3), while the TPMT*2 allele was not found in any of the examined samples. TPMT enzyme activity distribution in the subgroup sample was bimodal and as such correlated with genetic data. Using a cutoff value of 9.82 pmol/10(7) RBC per h, the genetic data correctly predicted TPMT enzyme activity in 91.6% of the examined individuals. Pharmacogenetic TPMT analyses have therefore proved to have significant clinical implications for prediction of individuals' responses to treatment with thiopurine drugs in order to avoid possible life-threatening therapy-related toxicities.

Simulation study of granular compaction dynamics under vertical tapping.

We study, by numerical simulation, the compaction dynamics of frictional hard disks in two dimensions, subjected to vertical shaking. Shaking is modeled by a series of vertical expansion of the disk packing, followed by dynamical recompression of the assembly under the action of gravity. The second phase of the shake cycle is based on an efficient event-driven molecular-dynamics algorithm. We analyze the compaction dynamics for various values of friction coefficient and coefficient of normal restitution. We find that the time evolution of the density is described by rho(t)=rho{infinity}-DeltarhoE{alpha}[-(ttau){alpha}], where E{alpha} denotes the Mittag-Leffler function of order 0

Symmetry effects in reversible random sequential adsorption on a triangular lattice.

Reversible random sequential adsorption of objects of various shapes on a two-dimensional triangular lattice is studied numerically by means of Monte Carlo simulations. The growth of the coverage rho(t) above the jamming limit to its steady-state value rho(infinity) is described by a pattern rho(t) = rho(infinity - deltarhoE(beta)[-(t/tau)beta], where E(beta) denotes the Mittag-Leffler function of order beta element of (0, 1). The parameter tau is found to decay with the desorption probability P_ according to a power law tau = AP_(-gamma). The exponent gamma is the same for all shapes, gamma = 1.29 +/- 0.01, but the parameter A depends only on the order of symmetry axis of the shape. Finally, we present the possible relevance of the model to the compaction of granular objects of various shapes.

A microplate assay for DNA damage determination (fast micromethod).

A rapid and convenient procedure for DNA damage determination in cell suspensions and solid tissues on single microplates was developed. The procedure is based on the ability of commercially available fluorochromes to interact preferentially with dsDNA in the presence of ssDNA, RNA, and proteins at high pH (>12.0), thus allowing direct measurements of DNA denaturation without sample handling or stepwise DNA separations. The method includes a simple and rapid 40-min sample lysis in the presence of EDTA, SDS, and high urea concentration at pH 10, followed by time-dependent DNA denaturation at pH 12.4 after NaOH addition. The time course and the extent of DNA denaturation is followed in a microplate fluorescence reader at room temperature for less than 1 h. The method requires only 30 ng DNA per single well and could conveniently be used whenever fast analysis of DNA integrity in small samples has to be done, e.g., in patients' lymphocytes after irradiation or chemotherapy (about 3000 cells per sample), in solid tissues or biopsies after homogenization (about 25 microg tissue per well), or in environmental samples for genotoxicity assessment.

Albuminuria, cardiovascular morbidity and mortality in diabetic and non-diabetic subjects in a rural general practice.

OBJECTIVES: The aim of this study was to estimate the frequency of albuminuria in patients with and without diabetes mellitus, answering the question: 'How relevant for general practice are epidemiological findings that albuminuria is a significant risk indicator for the development of cardiovascular syndromes?'. METHOD: The relationship between albuminuria and vascular disease was studied in two groups of subjects: one group consisted of 138 patients with diabetes mellitus and the other of 160 patients without diabetes, randomly selected from a stratified sample comparable with known diabetics by age, sex and profession. The groups were examined in the same way and mortality was followed over 5 years (1994-1998). RESULTS: Albuminuria was significantly more often present in diabetics (P < 0.0001). Peripheral arterial occlusive disease was diagnosed in 27.3% of patients with diabetes mellitus, and in 8.8% of the control group (P < 0.001). In the diabetic group 40 (29%) had microalbuminuria, but only 20 (12.5%) of the other group had the condition. In the presence of albuminuria, the incidence of arterial occlusive disease was 53.6%, but in those without it was only 9.8%. CONCLUSIONS: Our study has demonstrated that albuminuria is frequently present with diabetes mellitus, peripheral arterial occlusive disease, hypertension, coronary heart disease, hyperlipidaemia and in cases with increased concentration of blood fibrinogen. The albuminuria was significantly more frequent in those who died in the observed 5-year period after first examination. Although albuminuria indicates higher risks of cardiovascular morbidity and mortality in diabetics and in non-diabetics, it is necessary to study further how useful is this evidence for general practice.

For better data, better utilized.

In Nigeria a training programme was devised whereby staff from health centres could appreciate the value of reliable data as a means of raising the standard of services. A process of learning by doing enabled the participants to turn routine data into tools for management and the improvement of service quality.