Expression of urokinase plasminogen activator receptor in resting and activated bovine neutrophils

Changes in urokinase-plasminogen activator (u-PA) and u-PA receptor (u-PAR) expression at the protein and mRNA level in resting neutrophils and in neutrophils activated by phorbol myristate acetate (PMA) were examined. Low amounts of u-PA were found intracellularly or membrane-bound in resting neutrophils. However, incubation of resting neutrophils with purified exogenous u-PA (10 IU/ml) revealed extensive binding of u-PA to cell membranes. Excess amino-terminal fragment of the u-PA molecule, a proteolytically inactive fragment of u-PA (amino acids 1-135) blocked binding of exogenous u-PA to the cell membrane. These results, collectively, indicate that the binding of u-PA is specific and that resting neutrophils have unoccupied u-PA receptors on their cell membrane. Addition of PMA led to an increase (P < 0.01) in total cell-associated, membrane-bound u-PA activity and u-PA mRNA expression by bovine neutrophils. In contrast. PMA increased u-PAR mRNA levels but this was accompanied by a decrease (2.5-fold; P < 0.01) in free, unoccupied u-PA binding sites. No significant effects on total cell-associated or membrane-bound u-PA were found when neutrophils were treated with 4-phorbol 12,13 didecanoate, a phorbol ester that does not activate protein kinase C (PKC). Furthermore, addition of 1-(5-isoquinolinesylphonyl)-2-methlylpiperazine dihydrochloride (H-7), a potent PKC inhibitor, blocked the effect of PMA on total cell-associated u-PA activity. Thus, PKC plays a role in the modulation of u-PA and u-PAR by PMA in bovine neutrophils.     

Logic Based Abstractions of Real-Time Systems

When verifying concurrent systems described by transition systems, state explosion is one of the most serious problems. If quantitative temporal information (expressed by clock ticks) is considered, state explosion is even more serious. We present a notion of abstraction of transition systems, where the abstraction is driven by the formulae of a quantitative temporal logic, called qu-mu-calculus, defined in the paper. The abstraction is based on a notion of bisimulation equivalence, called , n-equivalence, where is a set of actions and n is a natural number. It is proved that two transition systems are , n-equivalent iff they give the same truth value to all qu-mu-calculus formulae such that the actions occurring in the modal operators are contained in , and with time constraints whose values are less than or equal to n. We present a non-standard (abstract) semantics for a timed process algebra able to produce reduced transition systems for checking formulae. The abstract semantics, parametric with respect to a set of actions and a natural number n, produces a reduced transition system , n-equivalent to the standard one. A transformational method is also defined, by means of which it is possible to syntactically transform a program into a smaller one, still preserving , n-equivalence.     

Effect of vitamin E supplementation on neutrophil function, milk composition and plasmin activity in dairy cows in a commercial herd

Fifty-six Holstein dairy cows from a commercial dairy herd in the Northern part of Greece were used to determine the effect of vitamin E supplementation on immune parameters, milk composition and milk quality. Cows were assigned to one of two experimental groups: control (no vitamin E supplementation) and vitamin E supplementation. Supplementation of vitamin E started 4 weeks prior to and continued up to 12 weeks after parturition. Supplementation included daily oral administration of vitamin E at 3000 i.u./cow prepartum and was reduced to 1000 i.u./cow post partum. Blood samples were collected weekly for 8 weeks starting 4 weeks before parturition, neutrophils were isolated and the following parameters were determined in neutrophils activated by phorbol myristate acetate: total cell-associated and membrane-bound urokinase plasminogen activator (u-PA) activity and superoxide production. Milk samples were collected weekly and fat, protein, lactose, somatic cell count (SCC), plasmin and plasminogen-derived activity were determined. Activated neutrophils isolated from cows that received supplemental vitamin E had higher (P<0.01) total and membrane-bound u-PA activities during the first 3 weeks after parturition and higher (P<0.01) superoxide production during week 1 prepartum and week 1 post partum compared with the corresponding values of activated neutrophils isolated from control cows. Vitamin E supplementation had no effect (P=0.28) on plasminogen-derived activity in milk. Milk obtained from cows that received supplemental vitamin E had SCC lower by 25% (P<0.05) and plasmin lower by 30% (P<0.01) than corresponding values in milk obtained from control cows. The reduction in plasmin as a result of vitamin E supplementation is very beneficial to the dairy industry because plasmin reduces the cheese-yielding capacity of milk, affects the coagulating properties of milk and its overall ability to withstand processing during cheesemaking. In conclusion, vitamin E supplementation had positive effects on the function of bovine neutrophils and milk quality in a commercial dairy herd.     

Reduction of immunoreactivity of bovine beta-lactoglobulin upon combined physical and proteolytic treatment

Bovine beta-lactoglobulin was hydrolyzed with trypsin or chymotrypsin before, during and after treatment at 600 MPa and pH 6.8 for 10 min at 30, 37 and 44 degrees C. The extent of beta-lactoglobulin hydrolysis under pressure was noticeably higher than at atmospheric pressure, particularly when chymotrypsin was used. Addition of proteases at ambient pressure to previously pressure-treated beta-lactoglobulin gave only a modest increase in proteolysis with respect to the untreated protein. Products of enzyme hydrolysis under pressure were separated by reverse-phase HPLC, and were found to be different from those obtained at atmospheric pressure when chymotrypsin was used. The residual immunochemical reactivity of the products of combined pressure-enzyme treatment was assessed on the unresolved hydrolysates by ELISA tests using polyclonal and monoclonal antibodies, and on individual hydrolytic fractions by Western Blotting using sera of paediatric patients allergic to whey proteins in cow milk. The immunoreactivity of the whole hydrolysates was related to their content of residual intact beta-lactoglobulin, and no immunochemical reactivity was found for all the products of chymotrypsin hydrolysis under pressure. The results indicate that chymotrypsin effectively hydrolysed hydrophobic regions of beta-lactoglobulin that were transiently exposed during the pressure treatments and that were not accessible in the native protein or in the protein that had been previously pressure treated.     

Variable-gradient generator for micro- and nano-HPLC

A new, simple device generates accurate nano- and microflow rate gradients from any conventional HPLC system. The core of the new device is represented by an electric-actuated, computer-controlled, multiposition HPLC valve. The valve hosts six reservoirs for as many different mobile-phase compositions of increasing strength. A low flow rate stream pushes the weakest solvent through the column as long as required and at the desired flow rate, until the chromatographic run is started. From this time on, the electric actuation allows one to select which reservoir will be on-line with the column and for how long, thus generating a specific solvent gradient, through a sequence of controlled segments of precise mobile-phase composition. This permits one not only to exactly reproduce the programmed slope but also to achieve different gradient shapes (i.e., linear, convex, concave) for different separation needs. The new device has proven to be reliable and reproducible even at the lowest flow rate tested (250 nL x min(-1)) and in different chromatographic conditions.     

A Combinatorial Approach to Elucidating Tribochemical Mechanisms

A new type of combinatorial tribological experiment is presented, which explores a series of tribological conditions, such as load and relative velocity, spatially separated as a library on one single sample. As an example, a library displaying the results of tribological testing of an additive under a series of different loads has been prepared and analyzed. The tribological information acquired during the testing has been correlated with spectroscopic information from the tribologically stressed surface. The use of imaging and small-area X-ray photoelectron spectroscopy has allowed the identification of the different tribologically stressed areas and the acquisition of detailed spectroscopic information. The composition and the thickness of the tribofilm were found to be dependent on the applied load. The use of the combinatorial approach shows the potential to greatly facilitate rapid characterization of new lubricant additives.     

An Analytical Model to Measure the Effectiveness of Safety Management Systems: Global Safety Improve Risk Assessment (G-SIRA) Method

The ever-increasing complexity of production systems, together with the need to obtain efficient processes with limited costs, has led companies to develop custom tools for process control and management. Even for risk assessment, the traditional models often are overcome by methods that are best suited to specific needs. In this context, the aim of this paper was to propose a new model, which we call the global safety improve risk assessment (G-SIRA). This model can classify risks and identify corrective actions that allow the best risk reduction at the lowest cost. The proposed model, which is based on improvements to previous research, uses the analytic hierarchy process approach to develop a valid and simple tool for risk management. The G-SIRA method has been tested in a real-world application, i.e., it was applied to all of the processes of a textile company, and the results were compared with those obtained from the classical approach failure mode, effects, and criticality analysis. The comparison clearly showed the effectiveness of the proposed model.     

Obstacle Modelling Oriented to Safe Motion Planning and Control for Planar Rigid Robot Manipulators

Trajectory planning and tracking are crucial tasks in any application using robot manipulators. These tasks become particularly challenging when obstacles are present in the manipulator workspace. In this paper a n-joint planar robot manipulator is considered and it is assumed that obstacles located in its workspace can be approximated in a conservative way with circles. The goal is to represent the obstacles in the robot configuration space. The representation allows to obtain an efficient and accurate trajectory planning and tracking. A simple but effective path planning strategy is proposed in the paper. Since path planning depends on tracking accuracy, in this paper an adequate tracking accuracy is guaranteed by means of a suitably designed Second Order Sliding Mode Controller (SOSMC). The proposed approach guarantees a collision-free motion of the manipulator in its workspace in spite of the presence of obstacles, as confirmed by experimental results.