Up-regulation of specific tyrosinase mRNAs in mouse melanomas with the c2j gene substituted for the wild-type tyrosinase allele: Utilization in design of syngeneic immunotherapy models

The expression of cell-specialization genes is likely to be changing in tumor cells as their differentiation declines. Functional changes in these genes might yield unusual peptide epitopes with anti-tumor potential and could occur without modification in the DNA sequence of the gene. Melanomas undergo a characteristic decline in melanization that may reflect altered contributions of key melanocytic genes such as tyrosinase. Quantitative reverse transcriptase–PCR of the wild-type (C) tyrosinase gene in transgenic (C57BL/6 strain) mouse melanomas has revealed a shift toward alternative splicing of the pre-mRNA that generated increased levels of the Δ1b and Δ1d mRNA splice variants. The spontaneous c2j albino mutation of tyrosinase (in the C57BL/6 strain) changes the pre-mRNA splicing pattern. In c2j/c2j melanomas, alternative splicing was again increased. However, while some mRNAs (notably Δ1b) present in C/C were obligatorily absent, others (Δ3 and Δ1d) were elevated. In c2j/c2j melanomas, the percentage of total tyrosinase transcripts attributable to Δ3 reached approximately 2-fold the incidence in c2j/c2j or C/C skin melanocytes. The percentage attributable to Δ1d rose to approximately 2-fold the incidence in c2j/c2j skin, and to 10-fold that in C/C skin. These differences provide a basis for unique mouse models in which the melanoma arises in skin grafted from a C/C or c2j/c2j transgenic donor to a transgenic host of the same or opposite tyrosinase genotype. Immunotherapy designs then could be based on augmenting those antigenic peptides that are novel or overrepresented in a tumor relative to the syngeneic host.

Structural basis for specificity and potency of a flavonoid inhibitor of human CDK2, a cell cycle kinase.

The central role of cyclin-dependent kinases (CDKs) in cell cycle regulation makes them a promising target for studying inhibitory molecules that can modify the degree of cell proliferation. The discovery of specific inhibitors of CDKs such as polyhydroxylated flavones has opened the way to investigation and design of antimitotic compounds. A novel flavone, (-)-cis-5,7-dihydroxyphenyl-8-[4-(3-hydroxy-1-methyl)piperidinyl] -4H-1-benzopyran-4-one hydrochloride hemihydrate (L868276), is a potent inhibitor of CDKs. A chlorinated form, flavopiridol, is currently in phase I clinical trials as a drug against breast tumors. We determined the crystal structure of a complex between CDK2 and L868276 at 2.33 angstroms resolution and refined to an Rfactor 20.3%. The aromatic portion of the inhibitor binds to the adenine-binding pocket of CDK2, and the position of the phenyl group of the inhibitor enables the inhibitor to make contacts with the enzyme not observed in the ATP complex structure. The analysis of the position of this phenyl ring not only explains the great differences of kinase inhibition among the flavonoid inhibitors but also explains the specificity of L868276 to inhibit CDK2 and CDC2.

Bioenergetic scaling: metabolic design and body-size constraints in mammals.

The cytosolic phosphorylation ratio ([ATP]/[ADP][P(i)]) in the mammalian heart was found to be inversely related to body mass with an exponent of -0.30 (r = 0.999). This exponent is similar to -0.25 calculated for the mass-specific O2 consumption. The inverse of cytosolic free [ADP], the Gibbs energy of ATP hydrolysis (delta G'ATP), and the efficiency of ATP production (energy captured in forming 3 mol of ATP per cycle along the mitochondrial respiratory chain from NADH to 1/2 O2) were all found to scale with body mass with a negative exponent. On the basis of scaling of the phosphorylation ratio and free cytosolic [ADP], we propose that the myocardium and other tissues of small mammals represent a metabolic system with a higher driving potential (a higher delta G'ATP from the higher [ATP]/[ADP][P(i)]) and a higher kinetic gain [(delta V/Vmax)/delta [ADP]] where small changes in free [ADP] produce large changes in steady-state rates of O2 consumption. From the inverse relationship between mitochondrial efficiency and body size we calculate that tissues of small mammals are more efficient than those of large mammals in converting energy from the oxidation of foodstuffs to the bond energy of ATP. A higher efficiency also indicates that mitochondrial electron transport is not the major site for higher heat production in small mammals. We further propose that the lower limit of about 2 g for adult endotherm body size (bumblebee-bat, Estrucan shrew, and hummingbird) may be set by the thermodynamics of the electron transport chain. The upper limit for body size (100,000-kg adult blue whale) may relate to a minimum delta G'ATP of approximately 55 kJ/mol for a cytoplasmic phosphorylation ratio of 12,000 M-1.

A valorização da função do objecto de design em contexto museológico : lógicas expositivas - a museologia do design industrial

The design objects give us a testimony of those who imagined, designed, developed, manufactured and used them. Each object, intentionally or not, portrays its own story, all the visible details are part of a decision taken by someone at some time of its chronology. The act of collecting objects, as well as private collections are the basis for the creation of museums as we know them today. Musealization - taking objects into a museum - means that one is restoring, preserving, enhancing some objects compared to others. And when restoring these objects, one is restoring their symbolic capacity, i.e. the fact that they tell a story, means you are restoring its message. In a museum, although out of context and deprived of most of the functions to which they had been designed for, the objects acquire other function(s), preserving their importance. Design museums give us the possibility to have a closer view of the objects, rather than just look at them, along with the pedagogical function. Thus presents a proposal for museography regarding industrial design, which is based on the appreciation of the function of anonymous design objects, based on expository logic, that takes the visitor to see, instead of just looking at objects, offering the possibility of interaction with the same, increasing the relationship between human being - object - museum, including groups with special needs, which are often forgotten in these exhibitions. This dissertation is a reflection and a projectual intervention on the design object in a museum, clarifying the concepts of object and museum, covering issues regarding the relevance of design museums, and culminating in the presentation of a museography project, where the function of an object prevails

Accident trend monitoring and exploratory analysis. Final report.

National Highway Traffic Safety Administration, National Center for Statistics and Analysis, Washington, D.C.

Multivariate statistical analysis of highway accident and highway conditions. Final report.

Transportation Department, Office of University Research, Washington, D.C.

Safety and environmental design considerations in the use of commercial electronic variable-message signage. Final report.

Mode of access: Internet.

Multidisciplinary accident analysis. Final report.

National Highway Traffic Safety Administration, Washington, D.C.

Accident avoidance evaluation of General Motors experimental safety vehicle. Final report.

National Highway Traffic Safety Administration, Washington, D.C.

Evaluation of design analysis procedures and acceptance criteria for roadside hardware. Volume III - evaluating pre-report 230 crash tests. Final report.

Federal Highway Administration, Safety Design Division, McLean, Va.

Safe geometric design for minicars. Final report.

Federal Highway Administration, Office of Research and Development, Washington, D.C.

Final design and implementation plan for evaluating the effectiveness of FMVSS 208: occupant crash protection. Tasks #4 & #5 report.

National Highway Traffic Safety Administration, Washington, D.C.

Final design and implementation plan for evaluating the effectiveness of FMVSS 301: fuel system integrity. Tasks #4 & #5 report.

National Highway Traffic Safety Administration, Washington, D.C.

Experimental design for evaluating the safety benefits of railroad advance warning signs. Final report.

Federal Highway Administration, Office of Research and Development, Washington, D.C.

Determination of the operational performance requirements for a roadside accident countermeasure warrant system. Volume II - technical documentation. Final report.

Federal Highway Administration, Office of Safety and Traffic Operations Research and Development, McLean, Va.