A New SiF–Dipropargyl Glycerol Scaffold as a Versatile Prosthetic Group to Design Dimeric Radioligands: Synthesis of the [18F]BMPPSiF Tracer to Image Serotonin Receptors

A novel SiX–dipropargyl glycerol scaffold (X: H, F, or ¹⁸F) was developed as a versatile prosthetic group that provides technical advantages for the preparation of dimeric radioligands based on silicon fluoride acceptor pre‐ or post‐labeling with fluorine‐18. Rapid conjugation with the prosthetic group takes place in microwave‐assisted click conjugation under mild conditions. Thus, a bivalent homodimeric SiX–dipropargyl glycerol derivatized radioligand, [¹⁸F]BMPPSiF, with enhanced affinity was developed by using click conjugation. High uptake of the radioligand was demonstrated in 5‐HT_1A receptor‐rich regions in the brain with positron emission tomography. Molecular docking studies (rigid protein–flexible ligand) of BMPPSiF and known antagonists (WAY‐100635, MPPF, and MefWAY) with monomeric, dimeric, and multimeric 5‐HT_1A receptor models were performed, with the highest G score obtained for docked BMPPSiF: ?6.766 as compared with all three antagonists on the monomeric model. Multimeric induced‐fit docking was also performed to visualize the comparable mode of binding under in vivo conditions, and a notably improved G score of ?8.455 was observed for BMPPSiF. These data directly correlate the high binding potential of BMPPSiF with the bivalent binding mode obtained in the biological studies. The present study warrants wide application of the SiX–dipropargyl glycerol prosthetic group in the development of ligands for imaging with enhanced affinity markers for specific targeting based on peptides, nucleosides, and lipids.     

Effects of caffeine on lipoprotein lipase gene expression during the adipocyte differentiation process

In this study, the effects of caffeine on lipoprotein lipase (LPL) gene expression were investigated in the 3T3-F442A preadipocyte cell line during the adipocyte differentiation process by determining LPL enzymatic activity and its messenger RNA (mRNA) level. The results demonstrate that caffeine acts on the gene expression of LPL, an early marker of adipocyte differentiation. It has a biphasic action: it increases gene expression in terms of mRNA when it is added to preadipocytes during the early stage of differentiation, but this is accompanied by a reduction of enzymatic activity. On the other hand, when caffeine is added for long periods during differentiation and/or when it is added to mature adipocytes, it induces marked inhibition of mRNA levels, correlated with a marked reduction of secreted enzymatic activity. The inhibitory effect of caffeine on LPL mRNA level can be reproduced by theophylline, a phosphodiesterase inhibitor, and by dibutyryl cyclic AMP, a non-metabolizable analog of cyclic AMP. However, the effect of caffeine and theophylline lasts longer than that of cyclic AMP, suggesting that a mechanism other than inhibition of cyclic AMP hydrolysis may be involved in the action of caffeine.     

Digital titration: Automated image acquisition and analysis of load and growth of Chlamydophila psittaci

Traditionally, the amount of infective chlamydiae in a given sample is determined by inoculating dilution series into cell cultures and physically counting chlamydial inclusions. This approach is time consuming, tedious, and error prone, mainly when dealing with high titers. Therefore, this paper describes a largely automated technique that was developed to standardize the determination of chlamydial load in vitro. Cells are fixed at 36 h post-inoculation and bacteria visualized using standard immunological detection methods. Consequently, for 81 microscopic fields, an image is recorded at the interpolated focal plane. These images are then automatically processed using an ImageJ plugin and the obtained results are imported into Excel to determine the number of inclusion forming units per mL in the sample. The main advantage of this technique is that no or minimal sample dilution is required, thus minimizing dilution errors. In addition, this technique was employed during the early, middle and late growth stages of the chlamydial developmental cycle and results correlated well (P < 0.01) with 16S rRNA values from previous experiments, thereby proving its suitability to follow chlamydial growth in vitro. The method described is highly suitable for high throughput titration of cell culture inoculated samples and assessment of possible antichlamydial effects of novel compounds throughout the chlamydial growth cycle. Microsc. Res. Tech., 2009. © 2009 Wiley-Liss, Inc.     

Linking the proteins--elucidation of proteome-scale networks using mass spectrometry

Proteomes are intricate. Typically, thousands of proteins interact through physical association and post-translational modifications (PTMs) to give rise to the emergent functions of cells. Understanding these functions requires one to study proteomes as "systems" rather than collections of individual protein molecules. The abstraction of the interacting proteome to "protein networks" has recently gained much attention, as networks are effective representations, that lose specific molecular details, but provide the ability to see the proteome as a whole. Mostly two aspects of the proteome have been represented by network models: proteome-wide physical protein-protein-binding interactions organized into Protein Interaction Networks (PINs), and proteome-wide PTM relations organized into Protein Signaling Networks (PSNs). Mass spectrometry (MS) techniques have been shown to be essential to reveal both of these aspects on a proteome-wide scale. Techniques such as affinity purification followed by MS have been used to elucidate protein-protein interactions, and MS-based quantitative phosphoproteomics is critical to understand the structure and dynamics of signaling through the proteome. We here review the current state-of-the-art MS-based analytical pipelines for the purpose to characterize proteome-scale networks.     

Indoor emissions of total and fluorescent supermicron particles during HOMEChem

Inhalation of particulate matter is associated with adverse health outcomes. The fluorescent portion of supermicron particulate matter has been used as a proxy for bioaerosols. The sources and emission rates of fluorescent particles in residential environments are not well-understood. Using an ultraviolet aerodynamic particle sizer (UVAPS), emissions of total and fluorescent supermicron particles from common human activities were investigated during the HOMEChem campaign, a test-house investigation of the chemistry of indoor environments. Human occupancy and activities, including cooking and mopping, were found to be considerable sources of indoor supermicron fluorescent particles, which enhanced the indoor particle concentrations by two orders of magnitude above baseline levels. The estimated total (fluorescent) mass emission rates for the activities tested were in the range of 4-30 (1-11) mg per person meal for cooking and 0.1-4.9 (0.05-4.7) mg/h for occupancy and mopping. Model calculations indicate that, once released, the dominant fate of coarse particles (2.5-10 micrometer in diameter) was deposition onto indoor surfaces, allowing for the possibility of subsequent resuspension and consequent exposures over durations much longer than the ventilation time scale. Indoor coarse particle deposition would also contribute to soiling of indoor surfaces.     

Yeast ecology in French cider and black olive natural fermentations

In this study, rDNA ITS restriction analysis was used to identify yeasts from two naturally fermented products: French ciders and black olives. In cider musts and bottled ciders, the PCR-RFLP method generated 15 different ITS/RFLP profiles for a total of 208 isolates. The predominant yeasts corresponded to Saccharomyces bayanus, Saccharomyces cerevisiae, Lachancea cidri and Dekkera anomala. Three identified species: Candida sake, Candida tropicalis and Kluyveromyces marxianus had never been described before in ciders. For the black olive fermentation, the method allowed for identification of 11 profiles for a total of 137 isolates. A sequential apparition of yeasts was observed with Pichia anomala, Candida boidinii and Debaryomyces etchellsii being the predominant species. Four isolates did not correspond to any known species based on the sequencing of the D1/D2 region of the 26S rRNA gene. By using the rDNA ITS method, valuable information on yeast population biodiversity and dynamics in the naturally fermented food products studied was obtained.     

Evaluating Learning and Interactions in a Multimedia Environment

An empirical study was undertaken to evaluate second language learning with a videodisc named Vi-Conte. The 78 subjects were post-secondary students and adults and belonged either to a control group, a video-control group or an experimental group. The research methodology is presented as well as analyses of learners' navigational patterns, strategies, gains in vocabulary items, changes in attitude and global evaluation of the videodisc. Suggestions for the development of adaptive learning environments are made based on these findings. This revised version was published online in July 2006 with corrections to the Cover Date.     

Generating shapes by analogies: An application to hearing aid design

3D shape modeling is a crucial component of rapid prototyping systems that customize shapes of implants and prosthetic devices to a patient’s anatomy. In this paper, we present a solution to the problem of customized 3D shape modeling using a statistical shape analysis framework. We design a novel method to learn the relationship between two classes of shapes, which are related by certain operations or transformation. The two associated shape classes are represented in a lower dimensional manifold, and the reduced set of parameters obtained in this subspace is utilized in an estimation, which is exemplified by a multivariate regression in this paper. We demonstrate our method with a felicitous application to the estimation of customized hearing aid devices.