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L Vanhamme

Publications and source records attributed to L Vanhamme.

At least 19 recordsLinked to original sources
PubMed2003

Quantification of the glycogen 13C-1 NMR signal during glycogen synthesis in perfused rat liver.

We studied glycogen synthesis from glucose in perfused livers of fed (n = 4) and 24 h starved (n = 7) rats. Glycogenolysis was inhibited by BAY R3401 (150 microM) and proglycosyn (100 microM). After 60 min, we replaced 99% (13)C-1 glucose by natural abundance glucose. This pulse-chase design allowed us to recognize residual ongoing futile glycogen turnover from the release of initially deposited (13)C-label, into the (13)C-free chase medium. Net residual turnover was less than 2 +/- 0.7% and 0.6 +/- 0.2% of 1-(13)C glycogen deposition rates of 0.31 +/- 0.04 and 0.99 +/- 0.04 micromol glucose g(-1) min(-1), in starved and fed livers, respectively. The 1-(13)C glycogen signal was monitored throughout the experiment with proton-decoupled (13)C NMR spectroscopy and analyzed in the time domain using AMARES. We noticed progressive line-broadening in any single experiment in the chase phase. One or a sum of two to three overlapping Lorentzians, with different exponential damping factors, were fitted to the signal. When the S/N was better than 40, the fit always delivered a small and a broad component. In the chase phase, the fit with a single Lorentzian resulted in a decline of glycogen signal by about 15 +/- 4 and 12 +/- 2% in starved and fed rats, respectively. This apparent decline in 1-(13)C glycogen signal could not be accounted for by the appearance of equivalent amounts of (13)C-labeled metabolites in the perfusate. The fit with a sum of two Lorentzians resulted in a decline of glycogen signal intensity of 7 +/- 5 and 5 +/- 3% in starved and fed rats, respectively, which reduced the apparent turnover to 8 +/- 9% and 6 +/- 4%, respectively. Quantification of the growing (13)C-1 glycogen signal requires a model function that accommodates changes in line shape throughout the period under study.

Algorithms↗
PubMed2002

Improved Lanczos algorithms for blackbox MRS data quantitation.

Magnetic resonance spectroscopy (MRS) has been shown to be a potentially important medical diagnostic tool. The success of MRS depends on the quantitative data analysis, i.e., the interpretation of the signal in terms of relevant physical parameters, such as frequencies, decay constants, and amplitudes. A variety of time-domain algorithms to extract parameters have been developed. On the one hand, there are so-called blackbox methods. Minimal user interaction and limited incorporation of prior knowledge are inherent to this type of method. On the other hand, interactive methods exist that are iterative, require user involvement, and allow inclusion of prior knowledge. We focus on blackbox methods. The computationally most intensive part of these blackbox methods is the computation of the singular value decomposition (SVD) of a Hankel matrix. Our goal is to reduce the needed computational time without affecting the accuracy of the parameters of interest. To this end, algorithms based on the Lanczos method are suitable because the main computation at each step, a matrix-vector product, can be efficiently performed by means of the fast Fourier transform exploiting the structure of the involved matrix. We compare the performance in terms of accuracy and efficiency of four algorithms: the classical SVD algorithm based on the QR decomposition, the Lanczos algorithm, the Lanczos algorithm with partial reorthogonalization, and the implicitly restarted Lanczos algorithm. Extensive simulation studies show that the latter two algorithms perform best.

Algorithms↗
PubMed2001

Control and function of the bloodstream variant surface glycoprotein expression sites in Trypanosoma brucei.

African trypanosomes escape the host immune response through a periodical change of their surface coat made of one major type of protein, the variant surface glycoprotein. From a repertoire of a thousand variant surface glycoprotein genes available, only one is expressed at a time, and this takes place in a specialised expression site itself selected from a collection of an estimated 20-30 sites. As the specialised expression sites are long polycistronic transcription units, the variant surface glycoprotein is co-transcribed with several other genes termed expression site-associated genes. How do the trypanosomes only use a single specialised expression site at a time? Why are there two dozen specialised expression sites? What are the functions of the other genes of these transcription units? We review the currently available answers to these questions.

Animals↗
PubMed2001

The VSG expression sites of Trypanosoma brucei: multipurpose tools for the adaptation of the parasite to mammalian hosts.

The variant surface glycoprotein (VSG) genes of Trypanosoma brucei are transcribed in telomeric loci termed VSG expression sites (ESs). Despite permanent initiation of transcription in most if not all of these multiple loci, RNA elongation is abortive except in bloodstream forms where full transcription up to the VSG occurs only in a single ES at a time. The ESs active in bloodstream forms are polycistronic and contain several genes in addition to the VSG, named ES-associated genes (ESAGs). So far 12 ESAGs have been identified, some of which are present only in some ESs. Most of these genes encode surface proteins and this list includes different glycosyl phosphatidyl inositol (GPI)-anchored proteins such as the heterodimeric receptor for the host transferrin (ESAG7/6), integral membrane proteins such as the receptor-like transmembrane adenylyl cyclase (ESAG4) and a surface transporter (ESAG10). An interesting exception is ESAG8, which may encode a cell cycle regulator involved in the differentiation of long slender into short stumpy bloodstream forms. Several ESAGs belong to multigene families including pseudogenes and members transcribed out of the ESs, named genes related to ESAGs (GRESAGs). However, some ESAGs (7, 6 and 8) appear to be restricted to the ESs. Most of these genes can be deleted from the active ES without apparently affecting the phenotype of bloodstream form trypanosomes, probably either due to the expression of ESAGs from 'inactive' ESs (ESAG7/6) or due to the expression of GRESAGs (in particular, GRESAGs4 and GRESAGs1). At least three ESAGs (ESAG7, ESAG6 and SRA) share the evolutionary origin of VSGs. The presence of these latter genes in ESs may confer an increased capacity of the parasite for adaptation to various mammalian hosts, as suggested in the case of ESAG7/6 and proven for SRA, which allows T. brucei to infect humans. Similarly, the existence of a collection of slightly different ESAG4s in the multiple ESs might provide the parasite with adenylyl cyclase isoforms that may regulate growth in response to different environmental conditions. The high transcription rate and high recombination level that prevail in VSG ESs may have favored the generation and/or recruitment in these sites of genes whose hyper-evolution allows adaptation to a larger variety of hosts.

Animals↗
PubMed2001

MR spectroscopy quantitation: a review of time-domain methods.

In this article an overview of time-domain quantitation methods is given. Advantages of processing the data in the measurement domain are discussed. The basic underlying principles of the methods are outlined and from them the situations under which these algorithms perform well are derived. Also an overview of methods to preprocess the data is given. In that respect, signal-to-noise and/or resolution enhancement, the removal of unwanted components and corrections for model imperfections are discussed.

Algorithms↗
PubMed2001

The filtering approach to solvent peak suppression in MRS: a critical review.

Suppressing the solvent peak is important in many applications of biomedical NMR spectroscopy in order to quantify the metabolites with a great accuracy. Among the postprocessing methods proposed in the literature, many deal with the concept of filtering. However, several proposals lack a theoretical perspective and some have not been explicitly applied to quantification problems. The present article is intended to bridge this gap: five methods are analyzed from a theoretical perspective. Subsequently the different methods are applied to the same set of data, and then the latter are quantified using the model fitting method AMARES. With our set, the scheme proposed by T. Sundin et al. (J. Magn. Reson. 139(2), 189-204 (1999)) proved to be the most reliable method.

Deuterium↗
PubMed2001

An update on antigenic variation in African trypanosomes.

African trypanosomes can spend a long time in the blood of their mammalian host, where they are exposed to the immune system and are thought to take advantage of it to modulate their own numbers. Their major immunogenic protein is the variant surface glycoprotein (VSG), the gene for which must be in one of the 20--40 specialized telomeric expression sites in order to be transcribed. Trypanosomes escape antibody-mediated destruction through periodic changes of the expressed VSG gene from a repertoire of approximately 1000. How do trypanosomes exclusively express only one VSG and how do they switch between them?

Animals↗
PubMed2000

A transcript encoding a proteasome beta-subunit and a zinc finger protein in Trypanosoma brucei brucei.

During the screening of a Trypanosoma brucei brucei (T. b. brucei) cDNA library constructed from bloodstream form mRNA, we identified a 2.3kb cDNA encoding a proteasome beta subunit (ORF1) and a putative zinc finger protein (ORF2). Northern blot analysis indicated the presence of a digenic transcript as well as the two individual messengers in both procyclic and bloodstream forms of the parasite. Southern blot analysis showed the relevant locus to be unique. ORF1 encoded a 22.7kDa protein sharing over 50% identity with the eukaryotic PRCE (aka beta5) proteasome beta subunit. This protein contained a beta amino acid signature and residues involved in the catalytic activity. Further phylogenetic analysis indicated that this subunit as well as those from other kinetoplastids could be confidentially assigned to extant eukaryotic subfamilies such as beta1, beta2, and beta5. ORF2 encoded a 14.6kDa putative zinc finger protein containing five repeats of a CCHC motif commonly present in retroviral nucleocapsid proteins as well as proteins involved in vertebrate embryogenesis.

Amino Acid Sequence↗
PubMed2000

A role for the dynamic acylation of a cluster of cysteine residues in regulating the activity of the glycosylphosphatidylinositol-specific phospholipase C of Trypanosoma brucei.

The glycosylphosphatidylinositol-specific phospholipase C or VSG lipase is the enzyme responsible for the cleavage of the glycosylphosphatidylinositol anchor of the variant surface glycoprotein (VSG) and concomitant release of the surface coat in Trypanosoma brucei during osmotic shock or extracellular acidic stress. In Xenopus laevis oocytes the VSG lipase was expressed as a nonacylated and a thioacylated form. This thioacylation occurred within a cluster of three cysteine residues but was not essential for catalytic activity per se. These two forms were also detected in trypanosomes and appeared to be present at roughly equivalent amounts. A reversible shift to the acylated form occurred when cells were triggered to release the VSG by either nonlytic acid stress or osmotic lysis. A wild type VSG lipase or a gene mutated in the three codons for the acylated cysteines were reinserted in the genome of a trypanosome null mutant for this gene. A comparative analysis of these revertant trypanosomes indicated that thioacylation might be involved in regulating enzyme access to the VSG substrate.

Acylation↗
PubMed2000

A single-stranded DNA-binding protein shared by telomeric repeats, the variant surface glycoprotein transcription promoter and the procyclin transcription terminator of Trypanosoma brucei.

In Trypanosoma brucei the genes are organised into long polycistronic transcription units and only three promoters for protein-encoding genes and a single terminator have been characterised. These promoters recruit a polI-like RNA polymerase for the transcription units encoding the two major stage-specific antigens of the parasite, the variant surface glycoprotein (VSG) of the bloodstream form and procyclin of the insect-specific procyclic form, while the terminator is that of a procyclin transcription unit. By deletional and mutational analysis we defined the two DNA sequences essential for the activity of the VSG promoter from a bloodstream form transcription unit and one of the functional elements of the procyclin terminator. These three short sequences are similar, and their C-rich strand binds the same protein of 40 kDa. In addition, this factor also binds to the C-rich strand of the telomeric repeats, the consensus target sequence being 5'-CCCTNN-3'. The factor-binding sequences are functionally interchangeable in chimeric promoter or terminator constructs, although additional elements are required for full activity.

Animals↗
PubMed2000

Frequency-selective quantification of biomedical magnetic resonance spectroscopy data.

In this paper the possibility of obtaining accurate estimates of parameters of selected peaks in the presence of unknown or uninteresting spectral features in biomedical magnetic resonance spectroscopy (MRS) signals is investigated. This problem is denoted by frequency-selective parameter estimation. A new time-domain technique based on maximum-phase finite impulse response (FIR) filters is presented. The proposed method is compared to a number of existing approaches: the application of a weighting function in the time domain, frequency domain fitting using a polynomial baseline, and the time-domain HSVD filter method. The ease of use and low computational complexity of the FIR filter method make it an attractive approach for frequency-selective parameter estimation. The methods are validated using simulations of relevant (13)C and (31)P MRS examples.

Adenosine Triphosphate↗
PubMed2000

Differential RNA elongation controls the variant surface glycoprotein gene expression sites of Trypanosoma brucei.

The protozoan parasite Trypanosoma brucei develops antigenic variation to escape the immune response of its host. To this end, the trypanosome genome contains multiple telomeric expression sites competent for transcription of variant surface glycoprotein genes, but as a rule only a single antigen is expressed at any time. We used reverse transcription-PCR (RT-PCR) to analyse transcription of different segments of the expression sites in different variant clones of two independent strains of T. brucei. The results indicated that RNA polymerase is installed and active at the beginning of many, if not all, expression sites simultaneously, but that a progressive arrest of RNA elongation occurs in all but one site. This defect is linked to inefficient RNA processing and RNA release from the nucleus. Therefore, functional transcription in the active site appears to depend on the selective recruitment of a RNA elongation/processing machinery.

Animals↗
PubMed1999

Differential regulation of ESAG transcripts in Trypanosoma brucei.

In Trypanosoma brucei, several genes termed ESAGs for expression site-associated genes are contained within the polycistronic transcription units of the VSG gene, and their transcription is coordinately regulated during the parasite life-cycle. Since the VSG mRNA is characterized by a drastic destabilization under conditions where translation is inhibited, we examined if this post-transcriptional control also applies to the ESAG mRNAs. While the ESAG 7/6 mRNA behaved like the VSG mRNA, the ESAG 8 and ESAG 3 mRNAs did not. We ascribe this differential behaviour to the residual transcription that still occurs only in the ESAG 7/6 region of the VSG unit under conditions where this unit is down-regulated.

Animals↗
PubMed1999

Accurate quantification of (1)H spectra: from finite impulse response filter design for solvent suppression to parameter estimation.

A scheme for accurate quantification of (1)H spectra is presented. The method uses maximum-phase finite impulse response (FIR) filters for solvent suppression and an iterative nonlinear least-squares (NLLS) algorithm for parameter estimation. The estimation algorithm takes the filter influence on the metabolites of interest into account and can thereby correctly incorporate a large variety of prior knowledge into the estimation phase. The FIR filter is designed in such a way that no distortion of the important initial samples is introduced. The FIR filter method is compared numerically with the HSVD method for water signal removal in a number of examples. The results show that the FIR method, using an automatic filter design scheme, slightly outperforms the HSVD method in most cases. The good performance and ease of use of the FIR filter method combined with its low computational complexity motivate the use of the proposed method.

Algorithms↗
PubMed1999

Time-domain quantification of series of biomedical magnetic resonance spectroscopy signals.

Quantification of individual magnetic resonance spectroscopy (MRS) signals is possible in the time domain using interactive nonlinear least-squares fitting methods which provide maximum likelihood parameter estimates under certain assumptions or using fully automatic, but statistically suboptimal, black-box methods. In kinetic experiments time series of consecutive MRS spectra are measured in which information concerning the time evolution of some of the signal parameters is often present. The purpose of this paper is to show how AMARES, a representative example of the interactive methods, can be extended to the simultaneous processing of all spectra in the time series using the common information present in the spectra. We show that this approach yields statistically better results than processing the individual signals separately.

Adenosine Triphosphate↗
PubMed1998

A VSG expression site-associated gene confers resistance to human serum in Trypanosoma rhodesiense.

Infectivity of Trypanosoma brucei rhodesiense to humans is due to its resistance to a lytic factor present in human serum. In the ETat 1 strain this character was associated with antigenic variation, since expression of the ETat 1.10 variant surface glycoprotein was required to generate resistant (R) clones. In addition, in this strain transcription of a gene termed SRA was detected in R clones only. We show that the ETat 1.10 expression site is the one selectively transcribed in R variants. This expression site contains SRA as an expression site-associated gene (ESAG) and is characterized by the deletion of several ESAGs. Transfection of SRA into T.b. brucei was sufficient to confer resistance to human serum, identifying this gene as one of those responsible for T.b. rhodesiense adaptation to humans.

Animals↗