A growth-constrained environment drives tumor progression in vivo

We recently have shown that selective growth of transplanted normal hepatocytes can be achieved in a setting of cell cycle block of endogenous parenchymal cells. Thus, massive proliferation of donor-derived normal hepatocytes was observed in the liver of rats previously given retrorsine (RS), a naturally occurring alkaloid that blocks proliferation of resident liver cells. In the present study, the fate of nodular hepatocytes transplanted into RS-treated or normal syngeneic recipients was followed. The dipeptidyl peptidase type IV-deficient (DPPIV−) rat model for hepatocyte transplantation was used to distinguish donor-derived cells from recipient cells. Hepatocyte nodules were chemically induced in Fischer 344, DPPIV+ rats; livers were then perfused and larger (>5 mm) nodules were separated from surrounding tissue. Cells isolated from either tissue were then injected into normal or RS-treated DPPIV− recipients. One month after transplantation, grossly visible nodules (2–3 mm) were seen in RS-treated recipients transplanted with nodular cells. They grew rapidly, occupying 80–90% of the host liver at 2 months, and progressed to hepatocellular carcinoma within 4 months. By contrast, no liver nodules developed within 6 months when nodular hepatocytes were injected into the liver of recipients not exposed to RS, although small clusters of donor-derived cells were present in these animals. Taken together, these results directly point to a fundamental role played by the host environment in modulating the growth and the progression rate of altered cells during carcinogenesis. In particular, they indicate that conditions associated with growth constraint of the host tissue can drive tumor progression in vivo.

Betidamino acids: versatile and constrained scaffolds for drug discovery.

Betidamino acids (a contraction of "beta" position and "amide") are N'-monoacylated (optionally, N'-monoacylated and N-mono- or N,N'-dialkylated) aminoglycine derivatives in which each N'acyl/alkyl group may mimic naturally occurring amino acid side chains or introduce novel functionalities. Betidamino acids are most conveniently generated on solid supports used for the synthesis of peptides by selective acylation of one of the two amino functions of orthogonally protected aminoglycine(s) to generate the side chain either prior to or after the elongation of the main chain. We have used unresolved Nalpha-tert-butyloxycarbonyl-N'alpha-fluorenylmethoxycarbonyl++ + aminoglycine, and Nalpha-(Nalpha-methyl)-tert-butyloxycarbonyl-N'alpha-fluo renylmethoxycarbonyl aminoglycine as the templates for the introduction of betidamino acids in Acyline [Ac-D2Nal-D4Cpa-D3Pal-Ser-4Aph(Ac)-D4Aph(A c)-Leu-Ilys-Pro-DAla-NH2, where 2Nal is 2-naphthylalanine, 4Cpa is 4-chlorophenylalanine, 3Pal is 3-pyridylalanine, Aph is 4-aminophenylalanine, and Ilys is Nepsilon-isopropyllysine], a potent gonadotropin-releasing hormone antagonist, in order to test biocompatibility of these derivatives. Diasteremneric peptides could be separated in most cases by reverse-phase HPLC. Biological results indicated small differences in relative potencies (<5-fold) between the D and L nonalkylated betidamino acid-containing Acyline derivatives. Importantly, most betide diastereomers were equipotent with Acyline. In an attempt to correlate structure and observed potency, Ramachandran-type plots were calculated for a series of betidamino acids and their methylated homologs. According to these calculations, betidamino acids have access to a more limited and distinct number of conformational states (including those associated with alpha-helices, beta-sheets, or turn structures), with deeper minima than those observed for natural amino acids.

Potent, structurally constrained agonists and competitive antagonists of corticotropin-releasing factor.

Predictive methods, physicochemical measurements, and structure activity relationship studies suggest that corticotropin-releasing factor (CRF; corticoliberin), its family members, and competitive antagonists (resulting from N-terminal deletions) usually assume an alpha-helical conformation when interacting with the CRF receptor(s). To test this hypothesis further, we have scanned the whole sequence of the CRF antagonist [D-Phe12,Nle21,38]r/hCRF-(12-41) (r/hCRF, rat/human CRF; Nle, norleucine) with an i-(i + 3) bridge consisting of the Glu-Xaa-Xaa-Lys scaffold. We have found astressin [cyclo(30-33)[D-Phe12,Nle21,38,Glu30,Lys33]r/ hCRF(12-41)] to be approximately 30 times more potent than [D-Phe12,Nle21,38]r/hCRF-(12-41), our present standard, and 300 times more potent than the corresponding linear analog in an in vitro pituitary cell culture assay. Astressin has low affinity for the CRF binding protein and high affinity (Ki = 2 nM) for the cloned pituitary receptor. Radioiodinated [D-125I-Tyr12]astressin was found to be a reliable ligand for binding assays. In vivo, astressin is significantly more potent than any previously tested antagonist in reducing hypophyseal corticotropin (ACTH) secretion in stressed or adrenalectomized rats. The cyclo(30-33)[Ac-Pro4,D-Phe12,Nle21,38,Glu30,Lys33++ +]r/hCRF-(4-41) agonist and its linear analog are nearly equipotent, while the antagonist astressin and its linear form vary greatly in their potencies. This suggests that the lactam cyclization reinstates a structural constraint in the antagonists that is normally induced by the N terminus of the agonist.

Enhanced conformational sampling in Monte Carlo simulations of proteins: application to a constrained peptide.

A Monte Carlo simulation method for globular proteins, called extended-scaled-collective-variable (ESCV) Monte Carlo, is proposed. This method combines two Monte Carlo algorithms known as entropy-sampling and scaled-collective-variable algorithms. Entropy-sampling Monte Carlo is able to sample a large configurational space even in a disordered system that has a large number of potential barriers. In contrast, scaled-collective-variable Monte Carlo provides an efficient sampling for a system whose dynamics is highly cooperative. Because a globular protein is a disordered system whose dynamics is characterized by collective motions, a combination of these two algorithms could provide an optimal Monte Carlo simulation for a globular protein. As a test case, we have carried out an ESCV Monte Carlo simulation for a cell adhesive Arg-Gly-Asp-containing peptide, Lys-Arg-Cys-Arg-Gly-Asp-Cys-Met-Asp, and determined the conformational distribution at 300 K. The peptide contains a disulfide bridge between the two cysteine residues. This bond mimics the strong geometrical constraints that result from a protein's globular nature and give rise to highly cooperative dynamics. Computation results show that the ESCV Monte Carlo was not trapped at any local minimum and that the canonical distribution was correctly determined.

Architecture of a Bench-Type Carbonate Lake Margin and Its Relation to Fluvially Dominated Deltas, Las Minas Basin, Upper Miocene, Spain

The Upper Miocene stratigraphic succession of the Las Minas Basin, located at the external zone of the Betic Chain in SE Spain, preserves several examples of lake carbonate bench deposits. Excellent exposures of the carbonate benches allow detailed observation of the architecture of these sediments and provide new insights for the ‘‘steep-gradient bench margin–low energy’’ model proposed by Platt and Wright (1991). The lake carbonate benches developed in close association with fluvially dominated shallow deltas that exhibit typical Gilbert-type profiles. The delta sequences comprise bottomset prodelta marl facies, distal to proximal foreset facies, deposited mainly in a delta-front environment, and topset facies, the latter reflecting both subaqueous delta-front and subaerial delta-plain environments. The development of the carbonate benches was constrained by the convexupward morphology of the deltaic deposits, which led to the available accommodation space for the growth of the steep-gradient platforms. The benches display a progradational pattern characterized by sigmoid-oblique internal geometries and offlap upper boundary relationships, which suggests that the carbonate benches developed under slow though continuous lake-level rise. Both the dimensions of the benches and the dominant carbonate components (i.e., encrusted charophyte stems and calcified cyanobaterial remains), allow comparisons with the progradational marl benches recognized in modern temperate hardwater lakes. Accordingly, the case study presented here provides a good ancient sedimentary analog for low-energy lake carbonate benches. Moreover, the evolutionary trend inferred from the fossil example offers new insights into the depositional conditions of this type of sediment and allows recognition of the transitional pattern from bench to ramp carbonate lake margins.

Noise and PSRR improvement technique for TPC readout front-end in CMOS. technology.

ALICE is one of four major experiments of particle accelerator LHC installed in the European laboratory CERN. The management committee of the LHC accelerator has just approved a program update for this experiment. Among the upgrades planned for the coming years of the ALICE experiment is to improve the resolution and tracking efficiency maintaining the excellent particles identification ability, and to increase the read-out event rate to 100 KHz. In order to achieve this, it is necessary to update the Time Projection Chamber detector (TPC) and Muon tracking (MCH) detector modifying the read-out electronics, which is not suitable for this migration. To overcome this limitation the design, fabrication and experimental test of new ASIC named SAMPA has been proposed . This ASIC will support both positive and negative polarities, with 32 channels per chip and continuous data readout with smaller power consumption than the previous versions. This work aims to design, fabrication and experimental test of a readout front-end in 130nm CMOS technology with configurable polarity (positive/negative), peaking time and sensitivity. The new SAMPA ASIC can be used in both chambers (TPC and MCH). The proposed front-end is composed of a Charge Sensitive Amplifier (CSA) and a Semi-Gaussian shaper. In order to obtain an ASIC integrating 32 channels per chip, the design of the proposed front-end requires small area and low power consumption, but at the same time requires low noise. In this sense, a new Noise and PSRR (Power Supply Rejection Ratio) improvement technique for the CSA design without power and area impact is proposed in this work. The analysis and equations of the proposed circuit are presented which were verified by electrical simulations and experimental test of a produced chip with 5 channels of the designed front-end. The measured equivalent noise charge was <550e for 30mV/fC of sensitivity at a input capacitance of 18.5pF. The total core area of the front-end was 2300?m × 150?m, and the measured total power consumption was 9.1mW per channel.

Pion-kaon scattering amplitude constrained with forward dispersion relations up to 1.6 GeV

In this work we provide simple and precise parametrizations of the existing πK scattering data from threshold up to 1.6 GeV, which are constrained to satisfy forward dispersion relations as well as three additional threshold sum rules. We also provide phenomenological values of the threshold parameters and of the resonance poles that appear in elastic scattering.