Application of regulatory sequence analysis and metabolic network analysis to the interpretation of gene expression data

We present two complementary approaches for the interpretation of clusters of co-regulated genes, such as those obtained from DNA chips and related methods. Starting from a cluster of genes with similar expression profiles, two basic questions can be asked: 1. Which mechanism is responsible for the coordinated transcriptional response of the genes? This question is approached by extracting motifs that are shared between the upstream sequences of these genes. The motifs extracted are putative cis-acting regulatory elements. 2. What is the physiological meaning for the cell to express together these genes? One way to answer the question is to search for potential metabolic pathways that could be catalyzed by the products of the genes. This can be done by selecting the genes from the cluster that code for enzymes, and trying to assemble the catalyzed reactions to form metabolic pathways. We present tools to answer these two questions, and we illustrate their use with selected examples in the yeast Saccharomyces cerevisiae. The tools are available on the web (http://ucmb.ulb.ac.be/bioinformatics/rsa-tools/; http://www.ebi.ac.uk/research/pfbp/; http://www.soi.city.ac.uk/~msch/)

Diversity of Mycobacterium tuberculosis strains in Nairobi, Kenya

Setting: Tuberculosis (TB) patients attending 16 public health facilities in Nairobi, Kenya. Objective: To determine the Mycobacterium tuberculosis (M.tuberculosis) strain families circulating in Nairobi, Kenya. Methods: Sputum specimens from consecutive new and previously treated smear positive pulmonary TB patients were collected between February and August 2010 and cultured on Lowenstein9Jensen media. Spoligotyping was done on DNA extracted from the first isolate of each patient. The international spoligotype data base (SpolDB4) was used to group isolates into strain families. Results: Fourty seven different strain families were identified from 536 isolates. The principal groups were; CAS1_KILI 96/536 (17%), T1 69/536 (12%), Beijing 65/536 (12%), LAM9 46/536 (9% ), LAM3 & S/Conversant 37/536 (7% ), LAM11_ZWE 26/536 (5%), CAS1_DELHI 24/536 (4%) and T2 24/536 (4%). Others identified and are found in the SpolDB4 were 113/536 (21%). A possible new M.tuberculosis strain family was identified with 21/536 (4%) isolates which was designated as Nairobi subtype. Others identified not previously included in the SpolDB4 accounted for 15/536 (3%). Conclusion: We found a diverse array of M.tuberculosis strain families which could be indicative of a cosmopolitant polulation with frequent migration that may suggest that the dorminant strain families may have been present in the population for an extended period of time or on going transmision of closely related strains families. The emergence of the Beijing strains poses a serious threat to TB control due to its high virulence and frequent association with multidrug resistance. We therefore call for strenghthening efforts on early case finding through enhanced public health education campains and provision of accessible diagnostic services with enhanced treatment compliance

Mycobacterium tuberculosis complex genetic diversity: mining the fourth international spoligotyping database (SpolDB4) for classification, population genetics and epidemiology

BACKGROUND: The Direct Repeat locus of the Mycobacterium tuberculosis complex (MTC) is a member of the CRISPR (Clustered regularly interspaced short palindromic repeats) sequences family. Spoligotyping is the widely used PCR-based reverse-hybridization blotting technique that assays the genetic diversity of this locus and is useful both for clinical laboratory, molecular epidemiology, evolutionary and population genetics. It is easy, robust, cheap, and produces highly diverse portable numerical results, as the result of the combination of (1) Unique Events Polymorphism (UEP) (2) Insertion-Sequence-mediated genetic recombination. Genetic convergence, although rare, was also previously demonstrated. Three previous international spoligotype databases had partly revealed the global and local geographical structures of MTC bacilli populations, however, there was a need for the release of a new, more representative and extended, international spoligotyping database. RESULTS: The fourth international spoligotyping database, SpolDB4, describes 1939 shared-types (STs) representative of a total of 39,295 strains from 122 countries, which are tentatively classified into 62 clades/lineages using a mixed expert-based and bioinformatical approach. The SpolDB4 update adds 26 new potentially phylogeographically-specific MTC genotype families. It provides a clearer picture of the current MTC genomes diversity as well as on the relationships between the genetic attributes investigated (spoligotypes) and the infra-species classification and evolutionary history of the species. Indeed, an independent Naïve-Bayes mixture-model analysis has validated main of the previous supervised SpolDB3 classification results, confirming the usefulness of both supervised and unsupervised models as an approach to understand MTC population structure. Updated results on the epidemiological status of spoligotypes, as well as genetic prevalence maps on six main lineages are also shown. Our results suggests the existence of fine geographical genetic clines within MTC populations, that could mirror the passed and present Homo sapiens sapiens demographical and mycobacterial co-evolutionary history whose structure could be further reconstructed and modelled, thereby providing a large-scale conceptual framework of the global TB Epidemiologic Network. CONCLUSION: Our results broaden the knowledge of the global phylogeography of the MTC complex. SpolDB4 should be a very useful tool to better define the identity of a given MTC clinical isolate, and to better analyze the links between its current spreading and previous evolutionary history. The building and mining of extended MTC polymorphic genetic databases is in progress

A theoretical consideration of the lysine-rich histones: H1 from a mammal and a echinoderm, H5 from erythrocytes

[No abstract available]Articl

Molecular answers to tuberculous questions

[No abstract available]Short Surve

Effects of adriamycin on heart and skeletal muscle chromatin

Interactions between adriamycin (ADM) and chromatin from heart and skeletal muscle from 15 day-old chicken emryos were investigated. Adriamycin interacts with the DNA of chromatin and this interaction is modified by the chromatin proteins. One of the effects of this interaction is an increase in the melting temperature (T(m)) of the DNA, where adriamycin is observed to increase the T(m) of heart chromatin to a greater extent than skeletal muscle chromatin. Adriamycin also inhibits in vitro DNA and RNA synthesis in isolated chromatin and nuclei. This inhibition is observed to be greater in heart muscle. Inhibition of transcription in the myocardium could be a major cause of adriamycin-induced cardiomyopathy.Articl

Preface

[No abstract available]Editoria

Inhibition of transcription by adriamycin is a consequence of the loss of negative superhelicity in DNA mediated by topoisomerase II

Adriamycin is commonly used as a chemotherapeutic agent and is known to intercalate into the major groove of DNA and inhibit DNA and RNA synthesis. Results presented in this communication suggest that adriamycin affects topoisomerase cleavage of DNA. The resultant change in negative superhelicity (decrease) is responsible for the decrease in transcription. This process is not dependent on the continued presence of adriamycin. The reaction between topoisomerases, DNA and adriamycin is dose-dependent. The results help to explain the relatively enhanced cytotoxicity of this drug to tumor cells.Articl

Adequacy of 6-month follow-up in chemotherapy trials

[No abstract available]Lette