Evolution of Ultracold, Neutral Plasmas

We present the first large-scale simulations of an ultracold, neutral plasma, produced by photoionization of laser-cooled xenon atoms, from creation to initial expansion, using classical molecular dynamics methods with open boundary conditions. We reproduce many of the experimental findings such as the trapping efficiency of electrons with increased ion number, a minimum electron temperature achieved on approach to the photoionization threshold, and recombination into Rydberg states of anomalously-low principal quantum number. In addition, many of these effects establish themselves very early in the plasma evolution ($\sim$ ns) before present experimental observations begin.Comment: 4 pages, 3 figures, submitted to PR

Diagnostic assays for leprosy based on T-cell epitopes.

To date, only a limited number of antigens have been described as specific for Mycobacterium leprae, and in many cases, homologues have subsequently been shown to exist in mycobacteria such as M. avium and M. intracellulare. A Leprosy Synthetic Peptide Skin Test Initiative was established by the Steering Committee on the Immunology of Mycobacteria of the UNDP/World Bank/WHO Special Programme for Research and Training in Tropical Diseases, to investigate the potential of synthetic peptides that encode T-cell epitopes as diagnostic tools, which could be used to develop a skin-test reagent specific for leprosy. Such M. leprae-specific peptides should have unique amino acid sequences, or significant sequence-dissimilarity from those in other mycobacteria. Synthetic peptides, 15 amino acids long, were synthesised from 33 genes or open reading frames within the M. leprae genome. Tuberculoid leprosy patients from four leprosy-endemic countries, Brazil, Ethiopia, Nepal and Pakistan, were tested as subjects known to have been infected with M. leprae, and to make good T-cell responses to antigens of M. leprae; UK blood donors were used as non-exposed or non-infected subjects. Peptides inducing potentially specific responses in leprosy patients and not in UK controls, and those inducing cross-reaction responses, present in both leprosy patients and non-exposed, non-infected controls, were identified. A difference from the equivalent M. tuberculosis sequence of five or more amino acid residues did not, by itself, identify peptides that were M. leprae-specific, suggesting that many of these peptides may have homologues in environmental mycobacteria. To date, this approach has identified a number of peptides with greater than 90% specificity and 19-47% sensitivity, which are undergoing further specificity-testing. Such peptides would have great potential as T-cell reagents with which to monitor exposure to M. leprae within communities, formulated either as skin-test reagents, or as antigens for tests in vitro

Competence to restart meiosis of oocytes from different follicle sizes obtained from slaughterhouse ovaries

Se evaluó la capacidad de los ovocitos de folículos según su tamaño para reiniciar la meiosis. Los folículos se midieron y clasificaron como Grupo 1 (G1 <4 mm), Grupo 2 (G2 4-8 mm) y Grupo 3 (G3 >8 mm). La aspiración se realizó por grupo con una aguja 21G conectada a una bomba de vacío, con una presión de 65 mmHg. Los complejos cúmulos de ovocitos (COC) recuperados se clasificaron como adecuados y no adecuados para la producción de embriones in vitro, de acuerdo con las características del cúmulo y del citoplasma. Los COC elegibles e inadecuados se valoraron con la prueba de azul brillante de Cresilo (BCB) y se clasificaron en BCB+ y BCB-. La maduración in vitro (IVM) se llevó a cabo en microgotas, incubadas en una cámara de CO2 al 5%, 38.5 °C y 90% de humedad durante 24 horas. La progresión meiótica se determinó por extrusión del corpúsculo polar mediante epifluorescencia bajo un microscopio invertido. La morfometría de ovocitos se estableció mediante una cámara de alta definición (Excelis AU-600-HD) y un software (AmScope v.3.7). El porcentaje de recuperación de ovocitos fue mayor de 63%. Los folículos de G2 proporcionaron un mayor porcentaje de COC elegibles (65.7%), donde el 59% de este grupo se clasificó como BCB+. Los ovocitos aptos de G1 y G2 reanudaron la meiosis en un 75%. Además, se observó que los ovocitos después de IVM redujeron su diámetro. Se concluye que los ovocitos de folículos entre 4-8 mm (G2) proporcionan un mayor porcentaje de COC maduros; sin embargo, el 50% de folículos <4 mm (G1) son una fuente prometedora de ovocitos viables, por lo que deben usarse para la producción de embriones in vitro.The competence of oocytes according to follicle size to restart meiosis was evaluated. The follicles were measured and classified as Group 1 (G1 <4 mm), Group 2 (G2 4-8 mm) and Group 3 (G3 >8 mm). The aspiration was performed by group with a 21G needle connected to a vacuum pump, with a pressure of 65 mmHg. The complex oocyte clusters (COCs) recovered were classified as suitable and unsuitable to produce embryos in vitro, according to the characteristics of the cumulus and the cytoplasm. Eligible and unsuitable COCs were assessed with the Brilliant Cresyl Blue (BCB) and were classified into BCB+ and BCB-. In vitro maturation (IVM) was carried out in microdroplets, incubated in a 5% CO2 chamber, 38.5 °C and 90% humidity for 24 hours. The meiotic progression was determined by extrusion of the polar corpuscle by epifluorescence under an inverted microscope. Oocyte morphometry was established using a high definition camera (Excelis AU-600-HD) and software (AmScope v.3.7). The recovery percentage of oocytes was greater than 63%. The G2 follicles provided a higher percentage of eligible COCs (65.7%), where 59% of this group was classified as BCB+. The fit oocytes of G1 and G2 resumed meiosis by 75%. In addition, it was observed that oocytes after IVM reduced their diameter. It is concluded that follicle oocytes between 4-8 mm (G2) provide a higher percentage of mature COCs; however, 50% of follicles <4 mm (G1) are a promising source of viable oocytes, so they should be used for in vitro embryo production

The mTOR inhibitor rapamycin down-regulates the expression of the ubiquitin ligase subunit Skp2 in breast cancer cells

INTRODUCTION: Loss of the cyclin-dependent kinase inhibitor p27 is associated with poor prognosis in breast cancer. The decrease in p27 levels is mainly the result of enhanced proteasome-dependent degradation mediated by its specific ubiquitin ligase subunit S phase kinase protein 2 (Skp2). The mammalian target of rapamycin (mTOR) is a downstream mediator in the phosphoinositol 3' kinase (PI3K)/Akt pathway that down-regulates p27 levels in breast cancer. Rapamycin was found to stabilize p27 levels in breast cancer, but whether this effect is mediated through changes in Skp2 expression is unknown. METHODS: The expression of Skp2 mRNA and protein levels were examined in rapamycin-treated breast cancer cell lines. The effect of rapamycin on the degradation rate of Skp2 expression was examined in cycloheximide-treated cells and in relationship to the anaphase promoting complex/Cdh1 (APC\C) inhibitor Emi1. RESULTS: Rapamycin significantly decreased Skp2 mRNA and protein levels in a dose and time-dependent fashion, depending on the sensitivity of the cell line to rapamycin. The decrease in Skp2 levels in the different cell lines was followed by cell growth arrest at G1. In addition, rapamycin enhanced the degradation rate of Skp2 and down-regulated the expression of the APC\C inhibitor Emi1. CONCLUSION: These results suggest that Skp2, an important oncogene in the development and progression of breast cancer, may be a novel target for rapamycin treatment

The Impact of Symbolic and Substantive Actions on Environmental Legitimacy

Drawing on institutional theory and insights from stakeholder theory and impression management, we empirically analyze the impact of both environmental symbolic polices (participation in voluntary environmental programs, green trademarks, environmental-dedicated board committees, environmental pay policies and community communication) and substantive actions (environmental patents and pollution prevention practices) on environmental legitimacy. We show that (1) symbolic actions have a weaker positive effect on legitimacy than substantive actions, (2) that the impact of symbolic actions is greater when they are combined with substantive actions, (3) that this impact is only short-term while substantive actions have both short- and long-term effects