Atomistic Insights into Rhodopsin Activation from a Dynamic Model

Rhodopsin, the light sensitive receptor responsible for blue-green vision, serves as a prototypical G protein-coupled receptor (GPCR). Upon light absorption, it undergoes a series of conformational changes that lead to the active form, metarhodopsin II (META II), initiating a signaling cascade through binding to the G protein transducin (G(t)). Here, we first develop a structural model of META II by applying experimental distance restraints to the structure of lumi-rhodopsin (LUMI), an earlier intermediate. The restraints are imposed by using a combination of biased molecular dynamics simulations and perturbations to an elastic network model. We characterize the motions of the transmembrane helices in the LUMI-to-META II transition and the rearrangement of interhelical hydrogen bonds. We then simulate rhodopsin activation in a dynamic model to study the path leading from LUMI to our META II model for wild-type rhodopsin and a series of mutants. The simulations show a strong correlation between the transition dynamics and the pharmacological phenotypes of the mutants. These results help identify the molecular mechanisms of activation in both wild type and mutant rhodopsin. While static models can provide insights into the mechanisms of ligand recognition and predict ligand affinity, a dynamic model of activation could be applicable to study the pharmacology of other GPCRs and their ligands, offering a key to predictions of basal activity and ligand efficacy.

Targeting treatment resistant breast cancer stem cells with FKBPL and its peptide derivative, AD-01, via the CD44 pathway

PURPOSE: FKBPL and its peptide derivative, AD-01, have already demonstrated tumour growth inhibition and CD44 dependent anti-angiogenic activity. Here we explore the ability of AD-01 to target CD44 positive breast cancer stem cells (BCSCs). EXPERIMENTAL DESIGN: Mammosphere assays and flow cytometry were utilized to analyse the effect of FKBPL overexpression/knockdown and AD-01 treatment ± other anti-cancer agents on BCSCs using breast cancer cell lines (MCF-7/MDA-231/ZR-75), primary patient samples and xenografts. Delays in tumour initiation were evaluated in vivo. The anti-stem cell mechanisms were determined using clonogenic assays, qPCR and immunofluorescence. RESULTS: AD-01 treatment was highly effective at inhibiting the BCSC population by reducing mammosphere forming efficiency (MFE) and ESA+/CD44+/CD24- or ALDH+ cell subpopulations in vitro and tumour initiation in vivo. The ability of AD-01 to inhibit the self-renewal capacity of BCSCs was confirmed; mammospheres were completely eradicated by the third generation. The mechanism appears to be due to AD-01-mediated BCSC differentiation demonstrated by a significant decrease in the number of holoclones and an associated increase in meroclones/paraclones; the stem cell markers, Nanog, Oct4 and Sox2, were also significantly reduced. Furthermore, we demonstrated additive inhibitory effects when AD-01 was combined with the Notch inhibitor, DAPT. AD-01 was also able to abrogate a chemo- and radiotherapy induced enrichment in BCSCs. Finally, FKBPL knockdown led to an increase in Nanog/Oct4/Sox2 and an increase in BCSCs, highlighting a role for endogenous FKBPL in stem cell signalling. CONCLUSIONS: AD-01 has dual anti-angiogenic and anti-BCSC activity which will be advantageous as this agent enters clinical trial.

General regulations and orders, 1804

Bound with: The manual and platoon exercises, &c., &c. (51 p.) and: General orders and observations on the movement and field exercise of infantry (16 p.) Inscribed on front paste-down: L. Colonel Brock. Also inscribed: Maj. Genl T.A. Smith late of the 1st. Rifle Regt. Bookplate of Robert B. Taylor, Norfolk. On front flyleaf: lengthy, two-page transcription in ink, headed /Copy/Circular/ dated "Horse Guards, 25th May '07" and signed: "Harry Calvert Adjt. General.

General regulations and orders, 1804

Bound with: The manual and platoon exercises, &c., &c. (51 p.) and: General orders and observations on the movement and field exercise of infantry (16 p.) Inscribed on front paste-down: L. Colonel Brock. Also inscribed: Maj. Genl T.A. Smith late of the 1st. Rifle Regt. Bookplate of Robert B. Taylor, Norfolk. On front flyleaf: lengthy, two-page transcription in ink, headed /Copy/Circular/ dated "Horse Guards, 25th May '07" and signed: "Harry Calvert Adjt. General.

Beam-foil-excited auger transitions in neon

Energy spectra of electrons ejected from collisions between a carbon foil and Ne projectiles with energies between 1.4 and 20 MeV have been measured. Continuous and discrete electron energy distributions are observed. Auger transitions of foil-excited Ne have been studied. Using relativistic Dirac-Fock multiconfiguration calculations, most of the measured Auger transitions have been identified.

Subsidios cruzados en servicios públicos domiciliarios basados en el avalúo catastral

Este artículo presenta una primera propuesta de determinación de subsidios y contribuciones de tarifas de servicios públicos domiciliarios – SPD para la Capital colombiana, que no parta de una clasificación de grupos (estratos socioeconómicos). A partir de un ejercicio exploratorio, se diseña un esquema de pagos del servicio de acueducto a partir del avalúo catastral del inmueble en que habita cada hogar. El ejercicio establece el pago por unidad de consumo de agua, de manera que cada hogar en Bogotá destine una misma proporción de su gasto total al gasto en el servicio de acueducto y garantiza que el total de pagos cubran los costos de provisión del servicio.

Seasonal adjustment and cointegration

El documento examina el efecto de filtros de ajuste en el tamaño y poder de prueba de cointegración que usan los residuales como pruebas ADF y PP, mediante procedimientos MonteCarlo y una aplicación empírica. Nuestros resultados indican que el uso de filtros distorsiona el tamaño y reduce el poder de estas pruebas.

Issues of water supply and contemporary urban society: the case of Greater Amman, Jordan

Over the last two decades, Jordan has suffered a chronic water crisis, and is the tenth most water-scarce nation on Earth. Such water stress has been well illustrated in the case of Greater Amman, the capital, which has grown dramatically from a population of around 2000 in the 1920s, to 2.17 million today. One of the distinctive characteristics of the water supply regime of Greater Amman is that since 1987 it has been based on a system of rationing, with households receiving water once a week for various durations. Amman is highly polarized socio-economically, and by means of household surveys, both quantitative and qualitative, conducted in high- and low-income divisions of the city, a detailed empirical evaluation of the storage and use of water, the strategies used by households to manage water and overall satisfaction with water supply issues is provided in this paper, looking specifically at issues of social equity. The analysis demonstrates the social and economic costs of water rationing and consequent management to be high, as well as emphasizing that issues of water quality are of central importance to all consumers regardless of their socio-economic status within the city.