Quantitative Modeling of GRK-Mediated β2AR Regulation

We developed a unified model of the GRK-mediated β2 adrenergic receptor (β2AR) regulation that simultaneously accounts for six different biochemical measurements of the system obtained over a wide range of agonist concentrations. Using a single deterministic model we accounted for (1) GRK phosphorylation in response to various full and partial agonists; (2) dephosphorylation of the GRK site on the β2AR; (3) β2AR internalization; (4) recycling of the β2AR post isoproterenol treatment; (5) β2AR desensitization; and (6) β2AR resensitization. Simulations of our model show that plasma membrane dephosphorylation and recycling of the phosphorylated receptor are necessary to adequately account for the measured dephosphorylation kinetics. We further used the model to predict the consequences of (1) modifying rates such as GRK phosphorylation of the receptor, arrestin binding and dissociation from the receptor, and receptor dephosphorylation that should reflect effects of knockdowns and overexpressions of these components; and (2) varying concentration and frequency of agonist stimulation “seen” by the β2AR to better mimic hormonal, neurophysiological and pharmacological stimulations of the β2AR. Exploring the consequences of rapid pulsatile agonist stimulation, we found that although resensitization was rapid, the β2AR system retained the memory of the previous stimuli and desensitized faster and much more strongly in response to subsequent stimuli. The latent memory that we predict is due to slower membrane dephosphorylation, which allows for progressive accumulation of phosphorylated receptor on the surface. This primes the receptor for faster arrestin binding on subsequent agonist activation leading to a greater extent of desensitization. In summary, the model is unique in accounting for the behavior of the β2AR system across multiple types of biochemical measurements using a single set of experimentally constrained parameters. It also provides insight into how the signaling machinery can retain memory of prior stimulation long after near complete resensitization has been achieved.     

How Accurate are Paleoecological Reconstructions of Early Paleontological and Archaeological Sites?

Paleoecology allows construction of paleoenvironmental models, faunal changes and evolutionary trends of paleontological taxa using modern analogs. However, when linking modern analogs to paleontological taxa in paleoecological reconstruction, differential taxonomic preservation in the fossil record has to be taken into account. Paleontologists have known the biased nature of the fossil record since Efremov’s publication on taphonomy in 1940, yet many ecological models of habitats associated with hominins in paleontological and archaeological sites in Africa and elsewhere barely address the complexity of the fossil record. We use randomly sampled ungulates from modern biomes in a comparative taxonomic abundance to demonstrate how the combination of modern thanatocoenoses and taphocoenoses, when used in reference to habitat-specific biocoenosis, produce better inferences of past habitats in paleontological and archaeological sites than approaches currently used.     

CHROMOSOMAL DIFFERENTIATION IN CLARKIA DUDLEYANA

Snow , Richard . (U. California, Davis.) Chromosomal differentiation in Clarkia dudleyana. Amer. Jour. Bot. 47 (4) : 302—309. Illus. 1960.—Clarkia dudleyana (n=9) is a common, colonial annual of the early-summer California flora. Of 275 individuals, derived from 9 natural populations and their garden-grown representatives, 17.1% were heterozygous for reciprocal translocations. Supernumerary chromosomes were also found in about 2% of the plants examined. The translocation heterozygotes are not distributed regularly over the species range but are concentrated near the geographical center of distribution. Most of the populations contained none or only a few heterozygotes, but in one colony 69% of 42 plants sampled were heterozygous. Judging from the meiotic metaphase associations observed, at least 5 different chromosome arrangements are present at this locality. Hybrids between colonies have invariably been translocation heterozygotes, the largest association found in such hybrids being a chain of all 18 chromosomes (a potential ring of 18). No correlation is evident between geographical separation and degree of cytological differentiation. Heterozygotes with smaller rings of 4 or 6 chromosomes, whether from natural populations or resulting from interpopulation hybridization, are highly fertile owing to the regular alternate disjunction of the chromosomes of the rings. In the larger rings of 12 to 18 chromosomes, derived from interpopulation crosses, segregation is much more irregular and leads to high sterility. It is possible that at least in some localities the heterozygotes enjoy a selective advantage over their homozygous sibs. It is also postulated that homozygosity for a particular chromosome arrangement may be selectively favored in a certain habitat, as a result of a position effect attendant upon placing formerly non-linked genes in the same linkage group through reciprocal translocation. The high degree of chromosomal differentiation between some populations of this species suggests that the complex heterozygotes of Oenothera have arisen as a result of hybridization of cytologically differentiated races.     

Multipotent Cancer Stem Cells Derived from Human Malignant Peritoneal Mesothelioma Promote Tumorigenesis

During the progression of malignant peritoneal mesothelioma (MPeM), tumor nodules propagate diffusely within the abdomen and tumors are characterized by distinct phenotypic sub-types. Recent studies in solid organ cancers have shown that cancer stem cells (CSCs) play a pivotal role in the initiation and progression of tumors. However, it is not known whether tumorigenic stem cells exist and whether they promote tumor growth in MPeM. In this study, we developed and characterized a CSC model for MPeM using stably expandable tumorigenic stem cells derived from patient tumors. We found morphologically distinct populations of CSCs that divide asymmetrically or symmetrically in MPeM in vitro cell culture. The MPeM stem cells (MPeMSCs) express stem cell markers c-MYC, NES and VEGFR2 and in the presence of matrix components cells form colony spheres. MPeMSCs are multipotent, differentiate into neuronal, vascular and adipose progeny upon defined induction and the differentiating cells express lineage-specific markers such as TUBB3, an early neuronal marker; vWF, VEGFA, VEGFC and IL-8, endothelial markers; and PPARγ and FABP4, adipose markers. Xenotransplantation experiments using MPeMSCs demonstrated early tumor growth compared with parental cells. Limiting dilution experiments using MPeMSCs and endothelial lineage-induced cells derived from a single MPeMSC resulted in early tumor growth in the latter group indicating that endothelial differentiation of MPeMSCs is important for MPeM tumor initiation. Our observation that the MPeM tumors contain stem cells with tumorigenic potential has important implications for understanding the cells of origin and tumor progression in MPeM and hence targeting CSCs may be a useful strategy to inhibit malignant progression.