Morphology and function of human Leydig cells in vitro. Immunocytochemical and radioimmunological analyses

The aim of our study was to show whether the cells isolated from testes of patients underwent bilateral orchiectomy for prostatic cancer are able to grown in vitro, and if so, are functionally active. Immuncytochemistry was performed to show the functional status of human cultured cells. In detail, immunolocalization of luteinizing hormone receptors (LHR), mitochondria, and cytoskeletal elements was demonstrated. Moreover, radioimmunological assay was used to measure testosterone secretion by cultured Leydig cells. Using Nomarski interference contrast and fine immunofluorescence analysis the positive immunostaining for LHR was observed in almost all Leydig cells, however it was of various intensity in individual cells. Testosterone measurement revealed significant difference between testosterone secretion by hCG-stimulated and unstimulated Leydig cells (p<0.05). Moreover, testosterone levels were significantly higher in 24- and 48-hour-cultures than in those of 72 hrs (p<0.05). Morphological analysis of Leydig cells in culture revealed the presence of mononuclear and multinucleate cells. The latter cells occurred in both hCG-stimulated and unstimulated cultures. In Leydig cells labeled with a molecular marker MitoTtracker, an abundance of mitochondria and typical distribution of microtubules and microfilaments were observed irrespective of the number of nuclei within the cell, suggesting no functional differences between mono- and multinucleate human Leydig cells in vitro. Since the percentage of multinucleate cells was similar in both hCG-stimulated and unstimulated cultures (23.70% and 22.80%), respectively, the appearance of these cell population seems to be independent of hormonal stimulation

Differential Regulation of Macropinocytosis by Abi1/Hssh3bp1 Isoforms

Macropinocytosis, which is a constitutive cellular process of fluid and macromolecule uptake, is regulated by actin cytoskeleton rearrangements near the plasma membrane. Activation of Rac1, which is proposed to act upstream of the actin polymerization regulatory Wave 2 complex, has been found to correlate with enhanced macropinocytosis. One of the components of the Wave 2 complex is Abi1. Multiple, alternatively spliced isoforms of Abi1 are expressed in mammalian cells, but the functional significance of the various isoforms is unknown.. Because of alternative mRNA splicing, exon 10 is absent from isoform 3, precluding similar binding of activated Rac1. Both isoforms, however, bound to inactive Rac1 through the same non-exon 10 site. Thus, Abi1 isoform 3-containing Wave 2 complex exhibited a differential binding to activated vs. inactive Rac1, whereas isoform 2-containing Wave 2 complex bound activated or inactive Rac1 comparably.Based on these observations, we postulate that Abi1 isoforms differentially regulate macropinocytosis as a consequence of their different relative affinities for activated Rac1 in Wave 2 complex. These findings also raise the possibility that isoform-specific roles occur in other Abi1 functions

Mapping localized surface plasmons within silver nanocubes using cathodoluminescence hyperspectral imaging

Localized surface plasmons within silver nanocubes less than 50 nm in size are investigated using high resolution cathodoluminescence hyperspectral imaging. Multivariate statistical analysis of the multidimensional luminescence dataset allows both the identification of distinct spectral features in the emission and the mapping of their spatial distribution. These results show a 490 nm peak emitted from the cube faces, with shorter wavelength luminescence coming from the vertices and edges; this provides direct experimental confirmation of theoretical predictions

Measurement of shower development and its Moli\`ere radius with a four-plane LumiCal test set-up

A prototype of a luminometer, designed for a future e+e- collider detector, and consisting at present of a four-plane module, was tested in the CERN PS accelerator T9 beam. The objective of this beam test was to demonstrate a multi-plane tungsten/silicon operation, to study the development of the electromagnetic shower and to compare it with MC simulations. The Moli\`ere radius has been determined to be 24.0 +/- 0.6 (stat.) +/- 1.5 (syst.) mm using a parametrization of the shower shape. Very good agreement was found between data and a detailed Geant4 simulation.Comment: Paper published in Eur. Phys. J., includes 25 figures and 3 Table

Site occupancies in ternary C15 ordered Laves phases

Site occupancies in three C15-structured AB{sub 2}(X) Laves phases have been determined by Atom Location by CHanneling Enhanced MIcroanalysis (ALCHEMI). In NbCr{sub 2}(V), the results were consistent with exclusive site occupancies of Nb for the A sublattice and Cr and V for the B sublattice. The B-site occupancy of V is not expected from atom size effects alone. In NbCr{sub 2}(Ti), the results were consistent with Ti partitioning mostly to the A sites with some anti-site defects likely. In HfV{sub 2}(Nb), the results were consistent with Nb partitioning between the A and B sites. The results of the ALCHEMI analyses of these ternary C15 Laves phase materials will be discussed with respect to previously determined phase diagrams and first-principles total energy and electronic structure calculations

Performance of fully instrumented detector planes of the forward calorimeter of a Linear Collider detector

Detector-plane prototypes of the very forward calorimetry of a future detector at an e+e- collider have been built and their performance was measured in an electron beam. The detector plane comprises silicon or GaAs pad sensors, dedicated front-end and ADC ASICs, and an FPGA for data concentration. Measurements of the signal-to-noise ratio and the response as a function of the position of the sensor are presented. A deconvolution method is successfully applied, and a comparison of the measured shower shape as a function of the absorber depth with a Monte-Carlo simulation is given.Comment: 25 pages, 32 figures, revised version following comments from referee

Luminometer for the future International Linear Collider - simulation and beam test results

LumiCal will be the luminosity calorimeter for the proposed International Large Detector of the International Linear Collider (ILC). The ILC physics program requires the integrated luminosity to be measured with a relative precision on the order of 10e-3, or 10e-4 when running in GigaZ mode. Luminosity will be determined by counting Bhabha scattering events coincident in the two calorimeter modules placed symmetrically on opposite sides of the interaction point. To meet these goals, the energy resolution of the calorimeter must be better than 1.5% at high energies. LumiCal has been designed as a 30-layer sampling calorimeter with tungsten as the passive material and silicon as the active material. Monte Carlo simulation using the Geant4 software framework has been used to identify design elements which adversely impact energy resolution and correct for them without loss of statistics. BeamCal, covering polar angles smaller than LumiCal, will serve for beam tuning, luminosity optimisation and high energy electron detection. Secondly, prototypes of the sensors and electronics for both detectors have been evaluated during beam tests, the results of which are also presented here.Comment: Technology and Instrumentation in Particle Physics 2011, Chicago, IL, USA. Presented June 11, 2011, and submitted to Physics Procedi