Proceedings : a conference of the American Medical Informatics Association. AMIA Fall Symposium, 1997
Switching the development focus of computer-based instruction from the concerns of delivery techn... more Switching the development focus of computer-based instruction from the concerns of delivery technology to the fundamentals of instructional methodology, is a notion that has received increased attention among educational theorists and instructional designers over the last several years. Building upon this precept, a proposed methodology and computer support system is presented for distilling educational objectives into concept maps using strategies derived from cognitive theory. Our system design allows for a flexible and extensible architecture in which an educator can create instructional modules that encapsulate their teaching strategies, and mimics the adaptive behavior used by experienced instructors in teaching complex educational objectives.
Journal of physiology and pharmacology : an official journal of the Polish Physiological Society, 1994
Blood coagulation is initiated after blood is exposed to the transmembrane glycoprotein Tissue Fa... more Blood coagulation is initiated after blood is exposed to the transmembrane glycoprotein Tissue Factor (TF). The continuous flow reactor has been used to study the flow-dependent activation of factor X by the complex of TF and factor VIIa. The apparent Km value of X for TF-VIIa in vesicles of 16 nM is low compared to the value of > 600 nM on a macroscopic surface. At 17 to 70 fmoles TF-VIIa/cm2, the steady state Xa production rate was fairly insensitive to the enzyme density, which is in contrast with classical Michaelis-Menten kinetics. The X activation rate was linear with the cube root of the flow rate. These results show that X activation is limited by transport of factor X across a boundary layer towards the enzyme complex.
Switching the development focus of computer-based instruction from the concerns of delivery techn... more Switching the development focus of computer-based instruction from the concerns of delivery technology to the fundamentals of instructional methodology, is a notion that has received increased attention among educational theorists and instructional designers over the last several years. Building upon this precept, a proposed methodology and computer support system is presented for distilling educational objectives into concept maps using strategies derived from cognitive theory. Our system design allows for a flexible and extensible architecture in which an educator can create instructional modules that encapsulate their teaching strategies, and mimics the adaptive behavior used by experienced instructors in teaching complex educational objectives.
Planar-supported phopholipid bilayers formed by te adsorpton of vesicles are increasingly used in... more Planar-supported phopholipid bilayers formed by te adsorpton of vesicles are increasingly used in the investigation of lipK-ependent reactis. We have studied the way in which these bilayers are forned with phopholipid vesicles coaining the btranembrane protein Tssue Factor (TF). TF complexed with te senne protease, factor Vlla, is the primary initiator of bklod coagulation by way of activation of the zymogen factor X. TF has been shown to orient randomly on the inner and outer leaflets of vesicles. We used proteolytic digestion to produce vesicles in which the exracellular domain of TF is located on the inner leaflet These vesicles show no cofactor activity for factor Vila as a result of the inability of the extacellular domain of TF to bind Vila. After freeze/thawing, 50% of the cofactor activity was regained, indicating reorientation of the sequestered, inner leaflet TF. Adsorpion of these vesicles to the inner surface of glass microcapillaries results in a continuous phospholpid bilayer. The microcapillaries were perftsed with a solution of factors Vlla and X, and the effluent was monitored for factor Xa production, a sensifive measure of the activity of the TF-Vlla complex. For coatings produced with the digested vesicles, minimal TF-Vlla acfivity was observed, showing that the supported bilayer preserves the oientation of the leaflets in the vesicles, i.e., the outer leaflet of the vesicles forms the outer leaflet of the supported bilayer.
Blood coagulation is initiated after blood is exposed to the transmembrane glycoprotein Tissue Fa... more Blood coagulation is initiated after blood is exposed to the transmembrane glycoprotein Tissue Factor (TF). The continuous flow reactor has been used to study the flow-dependent activation of factor X by the complex of TF and factor VIIa. The apparent Km value of X for TF-VIIa in vesicles of 16 nM is low compared to the value of > 600 nM on a macroscopic surface. At 17 to 70 fmoles TF-VIIa/cm2, the steady state Xa production rate was fairly insensitive to the enzyme density, which is in contrast with classical Michaelis-Menten kinetics. The X activation rate was linear with the cube root of the flow rate. These results show that X activation is limited by transport of factor X across a boundary layer towards the enzyme complex.
... CA Hasselbacber, Evan Waxman, Lisa T. Galati, Paul B. Contino, JB Alexander ROSS,* and Willia... more ... CA Hasselbacber, Evan Waxman, Lisa T. Galati, Paul B. Contino, JB Alexander ROSS,* and William R. Laws* ... First, kAB*, the rate of excited-state complex dissociation, must be compared to kf,B; if it is less than k,,, the back reaction to re-form A* from B*-X will be less likely. ...
We have examined the environments of the three phenol rotamers about the C(α)-C(β) bond in tyrosi... more We have examined the environments of the three phenol rotamers about the C(α)-C(β) bond in tyrosinamide by fluorescence quenching. Steady-state acrylamide quenching yields a nonlinear stern-Volmer plot. With three distinct emitting species and no other information about the system, it is impossible to analyze the data due to the number of variables which have to be determined. We therefore reduced the number of variables by independently determining the fractional intensity and dynamic quenching constant for each rotamer through time-resolved fluorescence quenching studies. These parameters were then used to analyze the steady-state data for any contribution of static quenching. We conclude that the nonlinear Stern-Volmer plot for the quenching of tyrosinamide by acrylamide is a consequence of each rotamer having a distinct dynamic quenching constant and the presence of static quenching. The static quenching can be represented by either the sphere-of-action model involving two of the three rotamers or the ground-state complex model involving all three rotamers.
The first direct evidence for specific metal-binding sites in pure human and pure rabbit sex ster... more The first direct evidence for specific metal-binding sites in pure human and pure rabbit sex steroid-binding protein (SBP) is obtained using the luminescent lanthanide terbium. Terbium, a probe for calcium sites in proteins, provided protection of the SBP steroid-binding activity in diluted human serum samples equivalent to that provided by calcium. Pure SBP, first treated with ethylenediaminetetraacetate, was dialyzed against buffer containing TbCl3. After gel filtration to remove nonspecifically bound terbium, the protein was denatured in urea. The amount of protein-bound terbium was determined by luminescence enhancement of the lanthanide using the chelator dipicolinate, yielding four metal-binding sites per mole of dimer protein from both species.
Proceedings : a conference of the American Medical Informatics Association. AMIA Fall Symposium, 1997
Switching the development focus of computer-based instruction from the concerns of delivery techn... more Switching the development focus of computer-based instruction from the concerns of delivery technology to the fundamentals of instructional methodology, is a notion that has received increased attention among educational theorists and instructional designers over the last several years. Building upon this precept, a proposed methodology and computer support system is presented for distilling educational objectives into concept maps using strategies derived from cognitive theory. Our system design allows for a flexible and extensible architecture in which an educator can create instructional modules that encapsulate their teaching strategies, and mimics the adaptive behavior used by experienced instructors in teaching complex educational objectives.
Journal of physiology and pharmacology : an official journal of the Polish Physiological Society, 1994
Blood coagulation is initiated after blood is exposed to the transmembrane glycoprotein Tissue Fa... more Blood coagulation is initiated after blood is exposed to the transmembrane glycoprotein Tissue Factor (TF). The continuous flow reactor has been used to study the flow-dependent activation of factor X by the complex of TF and factor VIIa. The apparent Km value of X for TF-VIIa in vesicles of 16 nM is low compared to the value of > 600 nM on a macroscopic surface. At 17 to 70 fmoles TF-VIIa/cm2, the steady state Xa production rate was fairly insensitive to the enzyme density, which is in contrast with classical Michaelis-Menten kinetics. The X activation rate was linear with the cube root of the flow rate. These results show that X activation is limited by transport of factor X across a boundary layer towards the enzyme complex.
Switching the development focus of computer-based instruction from the concerns of delivery techn... more Switching the development focus of computer-based instruction from the concerns of delivery technology to the fundamentals of instructional methodology, is a notion that has received increased attention among educational theorists and instructional designers over the last several years. Building upon this precept, a proposed methodology and computer support system is presented for distilling educational objectives into concept maps using strategies derived from cognitive theory. Our system design allows for a flexible and extensible architecture in which an educator can create instructional modules that encapsulate their teaching strategies, and mimics the adaptive behavior used by experienced instructors in teaching complex educational objectives.
Planar-supported phopholipid bilayers formed by te adsorpton of vesicles are increasingly used in... more Planar-supported phopholipid bilayers formed by te adsorpton of vesicles are increasingly used in the investigation of lipK-ependent reactis. We have studied the way in which these bilayers are forned with phopholipid vesicles coaining the btranembrane protein Tssue Factor (TF). TF complexed with te senne protease, factor Vlla, is the primary initiator of bklod coagulation by way of activation of the zymogen factor X. TF has been shown to orient randomly on the inner and outer leaflets of vesicles. We used proteolytic digestion to produce vesicles in which the exracellular domain of TF is located on the inner leaflet These vesicles show no cofactor activity for factor Vila as a result of the inability of the extacellular domain of TF to bind Vila. After freeze/thawing, 50% of the cofactor activity was regained, indicating reorientation of the sequestered, inner leaflet TF. Adsorpion of these vesicles to the inner surface of glass microcapillaries results in a continuous phospholpid bilayer. The microcapillaries were perftsed with a solution of factors Vlla and X, and the effluent was monitored for factor Xa production, a sensifive measure of the activity of the TF-Vlla complex. For coatings produced with the digested vesicles, minimal TF-Vlla acfivity was observed, showing that the supported bilayer preserves the oientation of the leaflets in the vesicles, i.e., the outer leaflet of the vesicles forms the outer leaflet of the supported bilayer.
Blood coagulation is initiated after blood is exposed to the transmembrane glycoprotein Tissue Fa... more Blood coagulation is initiated after blood is exposed to the transmembrane glycoprotein Tissue Factor (TF). The continuous flow reactor has been used to study the flow-dependent activation of factor X by the complex of TF and factor VIIa. The apparent Km value of X for TF-VIIa in vesicles of 16 nM is low compared to the value of > 600 nM on a macroscopic surface. At 17 to 70 fmoles TF-VIIa/cm2, the steady state Xa production rate was fairly insensitive to the enzyme density, which is in contrast with classical Michaelis-Menten kinetics. The X activation rate was linear with the cube root of the flow rate. These results show that X activation is limited by transport of factor X across a boundary layer towards the enzyme complex.
... CA Hasselbacber, Evan Waxman, Lisa T. Galati, Paul B. Contino, JB Alexander ROSS,* and Willia... more ... CA Hasselbacber, Evan Waxman, Lisa T. Galati, Paul B. Contino, JB Alexander ROSS,* and William R. Laws* ... First, kAB*, the rate of excited-state complex dissociation, must be compared to kf,B; if it is less than k,,, the back reaction to re-form A* from B*-X will be less likely. ...
We have examined the environments of the three phenol rotamers about the C(α)-C(β) bond in tyrosi... more We have examined the environments of the three phenol rotamers about the C(α)-C(β) bond in tyrosinamide by fluorescence quenching. Steady-state acrylamide quenching yields a nonlinear stern-Volmer plot. With three distinct emitting species and no other information about the system, it is impossible to analyze the data due to the number of variables which have to be determined. We therefore reduced the number of variables by independently determining the fractional intensity and dynamic quenching constant for each rotamer through time-resolved fluorescence quenching studies. These parameters were then used to analyze the steady-state data for any contribution of static quenching. We conclude that the nonlinear Stern-Volmer plot for the quenching of tyrosinamide by acrylamide is a consequence of each rotamer having a distinct dynamic quenching constant and the presence of static quenching. The static quenching can be represented by either the sphere-of-action model involving two of the three rotamers or the ground-state complex model involving all three rotamers.
The first direct evidence for specific metal-binding sites in pure human and pure rabbit sex ster... more The first direct evidence for specific metal-binding sites in pure human and pure rabbit sex steroid-binding protein (SBP) is obtained using the luminescent lanthanide terbium. Terbium, a probe for calcium sites in proteins, provided protection of the SBP steroid-binding activity in diluted human serum samples equivalent to that provided by calcium. Pure SBP, first treated with ethylenediaminetetraacetate, was dialyzed against buffer containing TbCl3. After gel filtration to remove nonspecifically bound terbium, the protein was denatured in urea. The amount of protein-bound terbium was determined by luminescence enhancement of the lanthanide using the chelator dipicolinate, yielding four metal-binding sites per mole of dimer protein from both species.
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Papers by Paul Contino