Biomechanical properties of single chondrocytes and chondrons determined by micromanipulation and finite-element modelling

A chondrocyte and its surrounding pericellular matrix (PCM) are defined as a chondron. Single chondrocytes and chondrons isolated from bovine articular cartilage were compressed by micromanipulation between two parallel surfaces in order to investigate their biomechanical properties and to discover the mechanical significance of the PCM. The force imposed on the cells was measured directly during compression to various deformations and then holding. When the nominal strain at the end of compression was 50 per cent, force relaxation showed that the cells were viscoelastic, but this viscoelasticity was generally insignificant when the nominal strain was 30 per cent or lower. The viscoelastic behaviour might be due to the mechanical response of the cell cytoskeleton and/or nucleus at higher deformations. A finite-element analysis was applied to simulate the experimental force-displacement/time data and to obtain mechanical property parameters of the chondrocytes and chondrons. Because of the large strains in the cells, a nonlinear elastic model was used for simulations of compression to 30 per cent nominal strain and a nonlinear viscoelastic model for 50 per cent. The elastic model yielded a Young''s modulus of 14 ± 1 kPa (mean ± s.e.) for chondrocytes and 19 ± 2 kPa for chondrons, respectively. The viscoelastic model generated an instantaneous elastic modulus of 21 ± 3 and 27 ± 4 kPa, a long-term modulus of 9.3 ± 0.8 and 12 ± 1 kPa and an apparent viscosity of 2.8 ± 0.5 and 3.4 ± 0.6 kPa s for chondrocytes and chondrons, respectively. It was concluded that chondrons were generally stiffer and showed less viscoelastic behaviour than chondrocytes, and that the PCM significantly influenced the mechanical properties of the cells.     

Isotopic identification of the sources of lead contamination for white storks (Ciconia ciconia) in a marshland ecosystem (Doñana,S.W. Spain)

White storks (Ciconia ciconia) fed in contaminated waters resulting from the Aznacollar acid mining-sludge spillage into the R. Guadiamar, which feeds the eastern flank of the Guadalquivir marshes (Do?ana), S.W. Spain. The sludge was rich in a range of toxic elements, and in organic pollutants such as the aromatic amines. Storks did not exhibit elevated metals in their blood immediately following the accident, but chick blood collected the year following the accident showed genotoxic damage compared to the controls. In this study lead isotope analysis was used to assess if the storks had ingested sludge-derived contaminants. The sludge lead isotope ratio was distinct from that of the Do?ana sediments. The stork blood lead isotope ratios exactly matched that of the sludge. It was concluded that the storks had ingested sludge-derived contaminants. A detailed study of the lead contamination along the R. Guadiamar and the R. Guadalquivir (of which the Guadiamar is a tributary) was also conducted to place the white stork colony lead exposure in the context of the spatial contamination of the storks' habitat.     

Using survival analysis to investigate the effect of UV-C and heat treatment on storage rot of strawberry and sweet cherry

Ultraviolet light and heat treatment are proposed as alternative techniques for the use of chemicals to reduce the development of the spoilage fungi Botrytis cinerea and Monilinia fructigena on strawberry and sweet cherry, respectively, during storage. In order to investigate the effect of both physical techniques on microbial inactivation and on fruit quality, inoculated berries were subjected to different temperatures (40-48 degrees C) and UV-C doses (0.05-1.50 J/cm2). For each condition, 20 berries were used. After the treatment, fungal growth, visual damage (holes, stains) and fruit firmness were evaluated during a period of 10 days. The experimental data were analysed statistically using survival analysis techniques. Fungal growth on strawberries was significantly retarded using UV-C doses of 0.05 J/cm2 and higher. The same treatment had no significant effect when applied to cherries. The highest doses (1.00 and 1.50 J/cm2) had a negative effect on the calyx of the strawberry, causing browning and drying of the leaves. No beneficial effect of a low temperature treatment (40-48 degrees C) on the shelf life of strawberries was observed, but fungal development on cherries was retarded at temperatures of 45 and 48 degrees C. These temperatures caused severe damage on strawberries (soft stains, holes, decreased firmness), but had no influence on the quality of sweet cherries.     

Reflective grating interferometer: a folded reversal and shearing wave-front interferometer

The reflecting grating interferometer (RGI) is a folded and reversal wave-front interferometer sensitive only to asymmetrical aberrations such as third-order coma. The RGI can isolate and evaluate coma both in nearly collimated and in noncollimated beams. We propose a RGI with a different optical configuration that includes a lateral shearing in addition to folding and reversal operations. With lateral shear, the RGI also becomes sensitive to other terms of third-order aberrations such as defocusing, astigmatism, and spherical aberration. Optical path difference equations for interpreting interferograms and numerical simulations are presented to show how the interferometer works in the shearing configuration. Its potential applications are described and discussed.     

An infrastructure to support cooperation of knowledge-level agents on the semantic Grid

Can a Knowledge-Level layer be located in the Semantic Grid infrastructure? Is it possible to design an Agent Communication Language (ACL) which enables Knowledge-Level agents to cooperate in a geographically distributed Semantic Grid despite nodes’ failures or malfunctions? This paper tries to address the above Semantic Grid challenges presenting an agent-based Open Service Architecture which integrates geographically distributed agents in a Semantic Grid. The architecture is well integrated with standard Internet components and technologies and supports communication among Knowledge-Level agents. The role of agents is to retrieve, execute and compose available services providing more sophisticated instances of them. Inter-agent communication is realized by exploiting an advanced Agent Communication Language which supports a fault-tolerant anonymous interaction protocol and satisfies a set of well defined Knowledge-Level programming requirements. Here, we present the design of the architecture and of the Agent Communication Language as well as their implementation. The architecture is evaluated by means of several case studies which highlight the main feature of our proposal. The main advantage of our approach is to demonstrate that different issues, such as high level inter-agent communication and fault tolerance, can be successfully integrated in Grid infrastructures which provide Web Services maintaining a clean design of the architecture and a Knowledge-Level characterization.     

Examination of the kinetics of herpes simplex virus glycoprotein D binding to the herpesvirus entry mediator, using surface plasmon resonance

Previously, we showed that truncated soluble forms of herpes simplex virus (HSV) glycoprotein D (gDt) bound directly to a truncated soluble form of the herpesvirus entry mediator (HveAt, formerly HVEMt), a cellular receptor for HSV. The purpose of the present study was to determine the affinity of gDt for HveAt by surface plasmon resonance and to compare and contrast the kinetics of an expanded panel of gDt variants in binding to HveAt in an effort to better understand the mechanism of receptor binding and virus entry. Both HveAt and gDt are dimers in solution and interact with a 2:1 stoichiometry. With HveAt, gD1(306t) (from the KOS strain of HSV-1) had a dissociation constant (KD) of 3.2 x 10(-6) M and gD2(306t) had a KD of 1.5 x 10(-6) M. The interaction between gDt and HveAt fits a 1:1 Langmuir binding model, i.e., two dimers of HveAt may act as one binding unit to interact with one dimer of gDt as the second binding unit. A gD variant lacking all signals for N-linked oligosaccharides had an affinity for HveAt similar to that of gD1(306t). A variant lacking the bond from cysteine 1 to cysteine 5 had an affinity for HveAt that did not differ from that of the wild type. However, variants with double cysteine mutations that eliminated either of the other two disulfide bonds showed decreased affinity for HveAt. This result suggests that two of the three disulfide bonds of gD are important for receptor binding. Four nonfunctional gDt variants, each representing one functional domain of gD, were also studied. Mutations in functional regions I and II drastically decreased the affinity of gDt for HveAt. Surprisingly, a variant with an insertion in functional region III had a wild-type level of affinity for HveAt, suggesting that this domain may function in virus entry at a step other than receptor binding. A variant with a deletion in functional region IV [gD1(Delta290-299t)] exhibited a 100-fold enhancement in affinity for HveAt (KD = 3.3 x 10(-8) M) due mainly to a 40-fold increase in its kinetic on rate. This agrees with the results of other studies showing the enhanced ability of gD1(Delta290-299t) to block infection. Interestingly, all the variants with decreased affinities for HveAt exhibited decreased kinetic on rates but only minor changes in their kinetic off rates. The results suggest that once the complex between gDt and HveAt forms, its stability is unaffected by a variety of changes in gD.     

Linguistic and location effects in compliance with pesticide warning labels for amateur and professional users

Three studies explored amateur and professional users' compliance with pesticide warning labels. Professionals were classified as people working in a profession in which the use of pesticides is a necessary part of their job. Amateurs used pesticides only in their leisure time. The first study showed that the wording used affected perception of the appropriateness of hazard statements, one of the most effective variations being the use of the personal pronoun (statements beginning "You should..."). The location of warning information was also found to affect actual compliance: Compliance increased when warning information was presented in the directions for use section. A supplemental directive increased compliance only for professional users. In a final study, "best-case" and "worst-case" linguistic variations were combined with best-case and worst-case locations for safety information. Instruction statements using the personal pronoun and presented in the directions for use section resulted in the highest levels of compliance. The differences in compliance between amateur and professional users are interpreted within the framework of Rasmussen's (1986) distinction among skill-, rule-, and knowledge-based behavior. Actual or potential applications of this research include the design of warning labels and safety information.     

Decoding two-dimensional complex multicomponent separations by autocovariance function

A new method for decoding two-dimensional (2D) multicomponent separations based on the use of the 2D Autocovariance function (2D-ACVF) has been developed. Theoretical models of single component (SC) spot distributions in 2D separations, both random and structured, are developed as the basis for a nonlinear estimation of both sample and separation system parameters from experimental 2D separations. The number of SCs, the average spot size, the spot capacity, and the saturation factor can be evaluated in the case of random SC spot patterns. The procedure was validated by extensive numerical simulation under conditions close to those usually found in GC x GC or 2D-polyacrylamid gel electrophoresis of proteins. The worse precision degree was no greater than 10% in the case of maximum spot density. This imprecision was fully accounted for, and it seems acceptable owing to the intrinsic statistical character of the estimation method. Structured multicomponent 2D separations, where SCs are linked by linear relationships, give rise to specific structured patterns in 2D-ACVF plots from which the parameters (phase and frequency) of the structured SC sequences can be evaluated: the study of 2D-ACVF makes it possible to decode multicomponent 2D separation, that is, to determine the number, relative abundance, and structural similarities of the single components. Pertinent expressions of the theoretical 2D-ACVF were derived for simple cases, and a procedure for decoding cases of structured 2D separations was developed and applied. It was shown that 2D separations containing both random and structured patterns of SC spots give rise to 2D-EACVF, which is the superimposition of the two component parts. This feature allows one, in principle, to decode the two components. The relevance of these results for Giddings sample dimensionality and separation dimensionality and their effective experimental evaluation is discussed.     

A formal framework for connective stability of highly decentralized cooperative negotiations

Multiagent cooperative negotiation is a promising technique for modeling and controlling complex systems. Effective and flexible cooperative negotiations are especially useful for open complex systems characterized by high decentralization (which implies a low amount of exchanged information) and by dynamic connection and disconnection of agents. Applications include ad hoc network management, vehicle formation, and physiological model combination. To obtain an effective control action, the stability of the negotiation, namely the guarantee that an agreement will be eventually reached, is of paramount importance. However, the techniques usually employed for assessing the stability of a negotiation can be hardly applied in open scenarios. In this paper, whose nature is mainly theoretical, we make a first attempt towards engineering stable cooperative negotiations proposing a framework for their analysis and design. Specifically, we present a formal protocol for cooperative negotiations between a number of agents and we propose a criterion for negotiation stability based on the concept of connective stability. This is a form of stability that accounts for the effects of structural changes on the composition of a system and that appears very suitable for multiagent cooperative negotiations. To show its possible uses, we apply our framework for connective stability to some negotiations taken from literature.