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Mapping Experiments (mapping + experiment)
Selected AbstractsUncoupling proteins 2 and 3 interact with members of the 14.3.3 familyFEBS JOURNAL, Issue 9 2000Benoit Pierrat Uncoupling proteins (UCPs) are members of the superfamily of the mitochondrial anion carrier proteins (MATP). Localized in the inner membrane of the organelle, they are postulated to be regulators of mitochondrial uncoupling. UCP2 and 3 may play an important role in the regulation of thermogenesis and, thus, on the resting metabolic rate in humans. To identify interacting proteins that may be involved in the regulation of the activity of UCPs, the yeast two-hybrid system was applied. Segments of hUCP2 containing the hydrophilic loops facing the intermembrane space, or combinations of these, were used to screen an adipocyte activation domain (AD) fusion library. The 14.3.3 protein isoforms ,, ,, , were identified as possible interacting partners of hUCP2. Screening of a human skeletal muscle AD fusion library, on the other hand, yielded several clones all of them encoding the , isoform of the 14.3.3 family. Mapping experiments further revealed that all these 14.3.3 proteins interact specifically with the C-terminal intermembrane space domain of both hUCP2 and hUCP3 whereas no interactions could be detected with the C-terminal part of hUCP1. Direct interaction between UCP3 and 14.3.3 , could be demonstrated after in vitro translation by coimmunoprecipitation. When coexpressed in a heterologous yeast system, 14.3.3 proteins potentiated the inhibitory effect of UCP3 overexpression on cell growth. These findings suggest that 14.3.3 proteins could be involved in the targeting of UCPs to the mitochondria. [source] Determination of the inheritance pattern of hyperthelia in cattle by maximum likelihood analysis 1JOURNAL OF ANIMAL BREEDING AND GENETICS, Issue 6 2000M. Brka Summary A previously published data-set with observations on supernumerary teats (hyperthelia) in dual-purpose Simmental was reanalysed by maximum-likelihood. The data comprised 537 unrelated animals and 614 members of 27 paternal half-sib families with known phenotype of each sire. The frequency of hyperthelia was 58% in unrelated animals, 51% in families with unaffected sire, and 73% in families with affected sires. Six different cases of single-gene inheritance were considered. The highest log-likelihood was obtained for additive inheritance and for a recessive pattern with 100% penetrance for recessive homozygotes and 32% for both other genotypes. Estimates for the gene frequency of the favourable allele were 0.34 and 0.29, respectively. Simple dominance or recessiveness with full or incomplete penetrance could be excluded. The possibility of finding paternal half-sib families with a heterozygous sire as a resource for a mapping experiment seem to be good in German Simmental. Zusammenfassung Untersuchung des Erbganges für Hyperthelie beim Rind mittels Maximum-Likelihood-Analyse Schon früher veröffentliche Daten mit Beobachtungen zum Auftreten überzähliger Zitzen (Hyperthelie) bei Fleckviehtieren wurden einer Maximum-Likelihood-Analyse unterzogen. Das Material bestand aus 537 unverwandten Tieren und 614 Tieren, die aus 27 verschiedenen väterlichen Halbgeschwistergruppen stammten, wobei der Phänotyp des Vaters jeweils bekannt war. Die relative Häufigkeit überzähliger Zitzen betrug 58% bei unverwandten Tieren, 51% bei Nachkommen von nicht betroffenen Vätern und 73% bei Nachkommen von Vätern, die selbst überzählige Zitzen aufwiesen. Sechs verschiedene Möglichkeiten monogener Vererbung wurden untersucht. Die höchsten Werte für die log-likelihood ergaben sich für einen additiven Erbgang sowie für eine Variante mit 100% Penetranz für die rezessiv Homozygoten und jeweils 32% Penetranz für die beiden anderen Genotypen. Für das erwünschte Allel wurde in beiden Varianten eine ähnliche Frequenz von 34% bzw. 29% geschätzt. Einfache dominante oder rezessive Erbgänge, auch mit unvollständiger Penetranz, konnten ausgeschlossen werden. Die Aussicht, für Kartierungsexperimente geeignete väterlichen Halbgeschwisterfamilien zu finden, scheint für die Rasse Fleckvieh günstig zu sein. [source] Genetic mapping of quantitative trait loci for resistance to Haemonchus contortus in sheepANIMAL GENETICS, Issue 3 2009K. Marshall Summary This paper presents results from a mapping experiment to detect quantitative trait loci (QTL) for resistance to Haemonchus contortus infestation in merino sheep. The primary trait analysed was faecal worm egg count in response to artificial challenge at 6 months of age. In the first stage of the experiment, whole genome linkage analysis was used for broad-scale mapping. The animal resource used was a designed flock comprising 571 individuals from four half-sib families. The average marker spacing was about 20 cM. For the primary trait, 11 QTL (as chromosomal/family combinations) were significant at the 5% chromosome-wide level, with allelic substitution effects of between 0.19 and 0.38 phenotypic standard deviation units. In general, these QTL did not have a significant effect on faecal worm egg count recorded at 13 months of age. In the second stage of the experiment, three promising regions (located on chromosomes 1, 3 and 4) were fine-mapped. This involved typing more closely spaced markers on individuals from the designed flock as well as an additional 495 individuals selected from a related population with a deeper pedigree. Analysis was performed using a linkage disequilibrium,linkage approach, under additive, dominant and multiple QTL models. Of these, the multiple QTL model resulted in the most refined QTL positions, with resolutions of <10 cM achieved for two regions. Because of the moderate size of effect of the QTL, and the apparent age and/or immune status specificity of the QTL, it is suggested that a panel of QTL will be required for significant genetic gains to be achieved within industry via marker-assisted selection. [source] Essential role of C/EBP, in G-CSF-induced transcriptional activation and chromatin modification of myeloid-specific genesGENES TO CELLS, Issue 4 2008Satoshi Iida Granulocyte colony-stimulating factor (G-CSF) regulates the proliferation and differentiation of neutrophilic progenitor cells. Here, we investigated the roles of CCAAT/enhancer-binding protein (C/EBP), in the G-CSF-induced transcriptional activation and chromatin modification of the CCR2 and myeloperoxidase (MPO) genes in IL-3-dependent myeloid FDN1.1 cells. Chromatin immunoprecipitation (ChIP) and electrophoretic mobility shift assays revealed that G-CSF activates C/EBP, to bind target promoters. ChIP mapping experiments across the CCR2 and MPO genes showed that G-CSF induces histone H3 modifications: the acetylation of Lys9, trimethylation of Lys4 and trimethylation of Lys9. The distribution profile of the trimethylated Lys9 was distinct from that of the two other modifications. All the G-CSF-induced C/EBP, recruitment, transcriptional activation and histone modifications were reversed by re-stimulation with IL-3, and were abolished by short hairpin RNA (shRNA)-mediated knockdown of C/EBP,. These results indicate that C/EBP, is activated by G-CSF to bind target promoters, and plays critical roles in the transcriptional activation and dynamic chromatin modification of target genes during neutrophil differentiation. [source] Fine mapping and detection of the causative mutation underlying Quantitative Trait LociJOURNAL OF ANIMAL BREEDING AND GENETICS, Issue 5 2010E. Uleberg Summary The effect on power and precision of including the causative SNP amongst the investigated markers in Quantitative Trait Loci (QTL) mapping experiments was investigated. Three fine mapping methods were tested to see which was most efficient in finding the causative mutation: combined linkage and linkage disequilibrium mapping (LLD); association mapping (MARK); a combination of LLD and association mapping (LLDMARK). Two simulated data sets were analysed: in one set, the causative SNP was included amongst the markers, while in the other set the causative SNP was masked between markers. Including the causative SNP amongst the markers increased both precision and power in the analyses. For the LLD method the number of correctly positioned QTL increased from 17 for the analysis without the causative SNP to 77 for the analysis including the causative SNP. The likelihood of the data analysis increased from 3.4 to 13.3 likelihood units for the MARK method when the causative SNP was included. When the causative SNP was masked between the analysed markers, the LLD method was most efficient in detecting the correct QTL position, while the MARK method was most efficient when the causative SNP was included as a marker in the analysis. The LLDMARK method, combining association mapping and LLD, assumes a QTL as the null hypothesis (using LLD method) and tests whether the ,putative causative SNP' explains significantly more variance than a QTL in the region. Thus, if the putative causative SNP does not only give an Identical-By-Descent (IBD) signal, but also an Alike-In-State (AIS) signal, LLDMARK gives a positive likelihood ratio. LLDMARK detected less than half as many causative SNPs as the other methods, and also had a relatively high false discovery rate when the QTL effect was large. LLDMARK may however be more robust against spurious associations, because the regional IBD is largely corrected for by fitting a QTL effect in the null hypothesis model. [source] Peak capacity of ion mobility mass spectrometry: the utility of varying drift gas polarizability for the separation of tryptic peptidesJOURNAL OF MASS SPECTROMETRY (INCORP BIOLOGICAL MASS SPECTROMETRY), Issue 4 2004Brandon T. Ruotolo Abstract Ion mobility mass spectrometry (IM-MS) peptide mass mapping experiments were performed using a variety of drift gases (He, N2, Ar and CH4). The drift gases studied cover a range of polarizabilities ((0.2,2.6) × 10,24 cm3) and the peak capacities obtained for tryptic peptides in each gas are compared. Although the different gases exhibit similar peak capacities (5430 (Ar) to 7580 (N2)) in some cases separation selectivity presumably based on peptide conformers (or conformer populations), is observed. For example the drift time profiles observed for some tryptic peptide ions from aldolase (rabbit muscle) show a dependence on drift gas. The transmission of high-mass ions (m/z > 2000) is also influenced by increased scattering cross-section of the more massive drift gases. Consequently the practical peak capacity for IM-MS separation cannot be assumed to be solely a function of resolution and the ability of a gas to distribute signals in two-dimensional space; rather, peak capacity estimates must account for the transmission losses experienced for peptide ions as the drift gas mass increases. Copyright © 2004 John Wiley & Sons, Ltd. [source] Raman spectroscopic signature of life in a living yeast cellJOURNAL OF RAMAN SPECTROSCOPY, Issue 7 2004Yu-San Huang Abstract We have discovered a Raman spectroscopic signature that sharply reflects the metabolic activity of a mitochondrion in a living yeast cell. Raman mapping experiments on a GFP-labeled yeast cell showed that this signature originated exclusively from mitochondria. Addition of KCN caused a rapid decrease and subsequent disappearance of the signature followed by gradual changes of the phospholipid Raman bands, indicating that respiration was first inhibited by KCN and then lowered metabolic activity gradually deteriorated the double-membrane structure of a mitochondrion. We can now monitor the life and death of a single cell by time- and space-resolved Raman spectroscopy. Copyright © 2004 John Wiley & Sons, Ltd. [source] Bayesian analyses of admixture in wild and domestic cats (Felis silvestris) using linked microsatellite lociMOLECULAR ECOLOGY, Issue 1 2006R. LECIS Abstract Methods recently developed to infer population structure and admixture mostly use individual genotypes described by unlinked neutral markers. However, Hardy,Weinberg and linkage disequilibria among independent markers decline rapidly with admixture time, and the admixture signals could be lost in a few generations. In this study, we aimed to describe genetic admixture in 182 European wild and domestic cats (Felis silvestris), which hybridize sporadically in Italy and extensively in Hungary. Cats were genotyped at 27 microsatellites, including 21 linked loci mapping on five distinct feline linkage groups. Genotypes were analysed with structure 2.1, a Bayesian procedure designed to model admixture linkage disequilibrium, which promises to assess efficiently older admixture events using tightly linked markers. Results showed that domestic and wild cats sampled in Italy were split into two distinct clusters with average proportions of membership Q > 0.90, congruent with prior morphological identifications. In contrast, free-living cats sampled in Hungary were assigned partly to the domestic and the wild cat clusters, with Q < 0.50. Admixture analyses of individual genotypes identified, respectively, 5/61 (8%), and 16,20/65 (25,31%) hybrids among the Italian wildcats and Hungarian free-living cats. Similar results were obtained in the past using unlinked loci, although the new linked markers identified additional admixed wildcats in Italy. Linkage analyses confirm that hybridization is limited in Italian, but widespread in Hungarian wildcats, a population that is threatened by cross-breeding with free-ranging domestic cats. The total panel of 27 loci performed better than the linked loci alone in the identification of domestic and known hybrid cats, suggesting that a large number of linked plus unlinked markers can improve the results of admixture analyses. Inferred recombination events led to identify the population of origin of chromosomal segments, suggesting that admixture mapping experiments can be designed also in wild populations. [source] Integrated analytical approach in veal calves administered the anabolic androgenic steroids boldenone and boldione: urine and plasma kinetic profile and changes in plasma protein expressionPROTEINS: STRUCTURE, FUNCTION AND BIOINFORMATICS, Issue 17 2007Rosa Draisci Abstract Surveillance of illegal use of steroids hormones in cattle breeding is a key issue to preserve human health. To this purpose, an integrated approach has been developed for the analysis of plasma and urine from calves treated orally with a single dose of a combination of the androgenic steroids boldenone and boldione. A quantitative estimation of steroid hormones was obtained by LC-APCI-Q-MS/MS analysis of plasma and urine samples obtained at various times up to 36 and 24,h after treatment, respectively. These experiments demonstrated that boldione was never found, while boldenone ,- and ,-epimers were detected in plasma and urine only within 2 and 24,h after drug administration, respectively. Parallel proteomic analysis of plasma samples was obtained by combined 2-DE, MALDI-TOF-MS and ,LC-ESI-IT-MS/MS procedures. A specific protein, poorly represented in normal plasma samples collected before treatment, was found upregulated even 36,h after hormone treatment. Extensive mass mapping experiments proved this component as an N-terminal truncated form of apolipoprotein A1 (ApoA1), a protein involved in cholesterol transport. The expression profile of ApoA1 analysed by Western blot analysis confirmed a significant and time dependent increase of this ApoA1 fragment. Then, provided that further experiments performed with a growth-promoting schedule will confirm these preliminary findings, truncated ApoA1 may be proposed as a candidate biomarker for steroid boldenone and possibly other anabolic androgens misuse in cattle veal calves, when no traces of hormones are detectable in plasma or urine. [source] A novel multifunctional labeling reagent for enhanced protein characterization with mass spectrometryRAPID COMMUNICATIONS IN MASS SPECTROMETRY, Issue 24 2001Eric C. Peters Individual peptides with lysine at the C-terminus as well as protein tryptic digests were reacted with 2-methoxy-4,5-dihydro-1H -imidazole, converting lysine residues to their 4,5-dihydro-1H -imidazol-2-yl derivatives. The mass spectra of derivatized digests exhibit a greater number of more intense features than their underivatized counterparts, thus increasing the information obtained in peptide mapping experiments. Additionally, MS/MS spectra of the derivatized peptides are greatly simplified in comparison to their native species, yielding primarily an easily interpretable series of y-ions. Finally, this novel label also enables differential quantitation studies, as a stable isotopic form containing four deuterium atoms can readily be produced. Copyright © 2001 John Wiley & Sons, Ltd. [source] Frequency Map Variations in Squirrel Monkey Primary Auditory Cortex,THE LARYNGOSCOPE, Issue 7 2005Steven W. Cheung MD Abstract Objective: The goal of this work is to understand the neural basis for cortical representation of hearing in highly vocal primates to gain insights into the substrates for communication. Variation patterns in frequency representation among animals are incorporated into an explanatory model to reconcile heterogeneous observations. Study Design: Prospective. Methods: Thirty-four squirrel monkeys underwent microelectrode mapping experiments in primary auditory cortex (AI) using tone pip stimuli. Characteristic frequency (CF) was extracted from the excitatory frequency receptive field. Frequency maps were reconstructed using Voronoi-Dirichlet tessellation. The spatial locations (rostral vs. caudal) of highest CF isofrequency contours (minimum length 1 mm) and highest CF neuronal clusters on the temporal gyral surface were analyzed. Results: Isofrequency contours at least 1 mm long with CFs greater than 2.9 kHz (75% cases) are accessible on the temporal gyrus. Variability of the highest CF isofrequency contours accessible on the temporal gyrus has an interquartile range from 2.9 to 5.1 (mean 4.3) kHz. The highest CF isofrequency contours are located mainly in rostral AI, whereas the highest CF neuronal clusters flanking fully expressed isofrequency contours are equally distributed in rostral and caudal locations. Conclusions: Squirrel monkey AI frequency map variations are sizeable across animals and small within single animals (interhemispheric comparison). AI frequency map variations, modeled as translations and rotations relative to the lateral sulcus, are independent transfers. Caution must be exercised when interpreting nominal frequency map changes that are attributed to hearing loss and auditory learning effects. [source] Electronic structure and transport properties of quantum dotsANNALEN DER PHYSIK, Issue 5 2004M. Tews Abstract The subject of this paper are electronic properties of isolated quantum dots as well as transport properties of quantum dots coupled to two electronic reservoirs. Thereby special focus is put on the effects of Coulomb interaction and possible correlations in the quantum dot states. First, the regime of sequential tunneling to the reservoirs is investigated. It is shown that in case degenerate states participate in transport, the resonance positions in the differential conductance generally depend on temperature and the degree of degeneracy. This effect can be used to directly probe degeneracies in a quantum dot spectrum. A further effect, characteristic for sequential tunneling events, is the complete blocking of individual channels for transport. A generalisation of the well known spin blockade is found for correlated dot states transitions through which are not directly spin-forbidden. In the second part, the electronic structure of spherical quantum dots is calculated. In order to account for correlation effects, the few-particle Schrödinger equation is solved by an exact diagonalization procedure. The calculated electronic structure compares to experimental findings obtained on colloidal semiconductor nanocrystals by Scanning Tunneling Spectroscopy. It is found that the electric field induced by the tunneling tip is gives rise to a Stark effect which can break the spherical symmetry of the electronic ground state density which is in agreement with wave-function mapping experiments. The symmetry breaking depends on the competition between exchange energy and the Stark energy. Moreover, a systematic dependence on particle number is found for the excitation energies of optical transitions which explains recent experimental findings on self-organized quantum dots. In the last part, co-tunneling in the Coulomb blockade regime is studied. For this end the tunneling current is calculated up to the forth order perturbation theory in the tunnel coupling by a real-time Green's function approach for the non-equilibrium case. The differential conductance calculated for a quantum dot containing up to two interacting electrons shows complex signatures of the excitation spectrum which are explained by a combination of co-tunneling and sequential tunneling processes. Thereby the calculations show a peak structure within the Coulomb blockade regime which has also been observed in experiment. [source] Mapping the Specific Cytoprotective Interaction of Humanin with the Pro-apoptotic Protein BidCHEMICAL BIOLOGY & DRUG DESIGN, Issue 5 2007Jungyuen Choi Humanin is a short endogenous peptide, which can provide protection from cell death through its association with various receptors, including the pro-apoptotic Bcl-2 family proteins Bid, Bim, and Bax. By using NMR chemical shift mapping experiments, we demonstrate that the interaction between Humanin-derived peptides and Bid is specific, and we localize the binding site to a region on the surface of Bid, which includes residues from the conserved helical BH3 domain of the protein. The BH3 domain mediates the association of Bid with other Bcl-2 family members and is essential for the protein's cytotoxic activity. The data suggest that Humanin exerts its cytoprotective activity by engaging the Bid BH3 domain; this would hinder the association of Bid with other Bcl-2 family proteins, thereby mitigating its toxicity. The identification of a Humanin-specific binding site on the surface of Bid reinforces its importance as a direct modulator of programmed cell death, and suggests a strategy for the design of cytoprotective peptide inhibitors of Bid. [source] | ||||||||||||||||