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Sample records for analysis establishes mycobacterium

  1. Polyphasic taxonomic analysis establishes Mycobacterium indicus pranii as a distinct species.

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    Vikram Saini

    necessitated further analysis of MIP with more sensitive and segregating parameters to ascertain its precise taxonomic position as a new species. The analysis of MIP and its comparison with other mycobacterial reference strains based on cellular and biochemical features, growth characteristics and chemotaxonomic studies like FAME profiling confirmed that MIP is uniquely endowed with diverse metabolic attributes that effectively distinguishes it from all the closely related mycobacteria including M. intracellulare and M. chimaera. CONCLUSION: The results presented in this study coupled with the non-pathogenic nature and different biochemical and immunomodulatory properties of MIP affirm it as a distinct species belonging to M. avium complex (MAC. It is further proposed to use an earlier suggested name Mycobacterium indicus pranii for this newly established mycobacterial species. This study also exemplifies the growing need for a uniform, consensus based broader polyphasic frame work for the purpose of taxonomy and speciation, particularly in the genus Mycobacterium.

  2. Variable-Number Tandem-Repeat Analysis of Respiratory and Household Water Biofilm Isolates of "Mycobacterium avium subsp. hominissuis" with Establishment of a PCR Database.

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    Iakhiaeva, Elena; Howard, Susan T; Brown Elliott, Barbara A; McNulty, Steven; Newman, Kristopher L; Falkinham, Joseph O; Williams, Myra; Kwait, Rebecca; Lande, Leah; Vasireddy, Ravikiran; Turenne, Christine; Wallace, Richard J

    2016-04-01

    "Mycobacterium aviumsubsp.hominissuis" is an important cause of pulmonary disease. It is acquired from environmental sources, but there is no methodology for large population studies. We evaluated the potential of variable-number tandem-repeat (VNTR) analysis. Clinical and household biofilmM. aviumisolates underwent molecular identification. Testing for IS901was done to separateM. aviumsubsp.aviumfromM. aviumsubsp.hominissuis VNTR types were defined using VNTR loci, and subtyping was performed using 3'hsp65and internal transcribed spacer (ITS) sequencing. Forty-nine VNTR types and eight subtypes ofM. aviumsubsp.hominissuis(IS901negative) were identified among 416 isolates ofM. aviumfrom 121 patients and 80 biofilm sites. Of those types, 67% were found only among patient isolates, 11% only among household water isolates, and 23% among both. Of 13 VNTR types that included ≥4 patients, the majority (61.5%) represented geographic clustering (same city). Most VNTR types with multiple patients belonged to the same 3'hsp65sequence code (sequevar). A total of 44 isolates belonging to fourM. aviumsubsp.hominissuisVNTR types (8%), including three with the rare Mav-F ITS sequence and 0/8 subspecies, produced amplicons with IS901PCR primers. By sequencing, all 44 amplicons were not IS901but ISMav6, which was recently observed in Japan but had not been previously described among U.S. isolates. VNTR analysis ofM. aviumsubsp.hominissuisisolates is easier and faster than pulsed-field gel electrophoresis. Seven VNTR loci separated 417 isolates into 49 types. No isolates ofM. aviumsubsp.aviumwere identified. The distributions of the VNTR copy numbers, the allelic diversity, and the low prevalence of ISMav6 differed from the findings for respiratory isolates reported from Japan. PMID:26739155

  3. Tuberculosis:an experience from Mycobacterium smears and culture analysis

    Institute of Scientific and Technical Information of China (English)

    Zeehaida M; Siti Asma H; Siti Hawa H; Zaidah AR; Norbanee TH

    2009-01-01

    Objective:Simple tests like direct smear of the acid fast bacilli (AFB)and Mycobacterium culture could assist the diagnosis of tuberculosis.This study is aimed at reviewing the outcome of smears,culture results and con-tamination rate among specimens requested for AFB smear and Mycobacterium culture.Methods:Retrospec-tive laboratory data analysis requesting for Mycobacterium culture from January 2005 till December 2006 was done in a tertiary teaching hospital of Universiti Sains Malaysia,Kubang Kerian,Kelantan,Malaysia.Re-sults:Four hundred and sixty seven (36.6%)isolates grew from 1 277 specimens.Of these isolates,314 (67.2%)grew Mycobacterium tuberculosis,23 (4.9%)grew Mycobacterium other than tuberculosis and 38 (8.1%)grew contaminants.Among the M.tuberculosis cultures,165 (52.5%)had growth of more than 100 confluent colonies,whereas 39 cultures (12.4%)had growth of less than 19 colonies.Direct smear for AFB among smear positive cases showed presence of more than 50 bacilli /line in 231 (49.5%)cases and smear negative cases accounted for 63 (13.5%).Among smear positive cases,291 (94.5%)cultures grew Myco-bacterium species and another 17 (5.5%)cultures grew contaminants.In smear negative cases,32 (62.7%) cultures grew Mycobacterium species and 19 (37.3%)cultures grew contaminants.Conclusion:The results from data analysis of the Mycobacterium cultures should be critically utilized in order to review the laboratory performance and to improve its services in the future.Some of the data is also useful to the administrators of the hospital in terms of estimating the risk of occupational hazard faced by the health care workers.

  4. Local and regional re-establishment of cellular immunity during curative antibiotherapy of murine Mycobacterium ulcerans infection.

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    Teresa G Martins

    Full Text Available BACKGROUND: Buruli ulcer (BU is a neglected necrotizing disease of the skin, subcutaneous tissue and bone, caused by Mycobacterium ulcerans. BU pathogenesis is associated with mycolactone, a lipidic exotoxin with cytotoxic and immunosuppressive properties. Since 2004, the World Health Organization recommends the treatment of BU with a combination of rifampicin and streptomycin (RS. Histological analysis of human tissue samples suggests that such antibiotic treatment reverses the mycolactone-induced local immunosuppression, leading to increased inflammatory infiltrations and phagocytosis of bacilli. METHODOLOGY/PRINCIPAL FINDINGS: We used a mouse model of M. ulcerans footpad infection, followed by combined RS treatment. Time-lapsed analyses of macroscopic lesions, bacterial burdens, histology and immunohistochemistry were performed in footpads. We also performed CFU counts, histology and immunohistochemistry in the popliteal draining lymph nodes (DLN. We observed a shift in the cellular infiltrates from a predominantly neutrophilic/macrophagic to a lymphocytic/macrophagic profile in the infected footpads of antibiotic-treated mice. This shift occurred before the elimination of viable M. ulcerans organisms, which were ultimately eradicated as demonstrated by the administration of dexamethasone. This reduction of bacillary loads was accompanied by an increased expression of inducible nitric oxide synthase (NOS2 or iNOS. Predominantly mononuclear infiltrates persisted in the footpads during and after treatment, coincident with the long persistence of non-viable poorly stained acid-fast bacilli (AFB. We additionally observed that antibiotherapy prevented DLN destruction and lymphocyte depletion, which occurs during untreated experimental infections. CONCLUSIONS/SIGNIFICANCE: Early RS treatment of M. ulcerans mouse footpad infections results in the rapid elimination of viable bacilli with pathogen eradication. However, non-viable AFB persisted for

  5. Mycobacterium tuberculosis universal stress protein Rv2623 regulates bacillary growth by ATP-Binding: requirement for establishing chronic persistent infection.

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    Joshua E Drumm

    2009-05-01

    Full Text Available Tuberculous latency and reactivation play a significant role in the pathogenesis of tuberculosis, yet the mechanisms that regulate these processes remain unclear. The Mycobacterium tuberculosisuniversal stress protein (USP homolog, rv2623, is among the most highly induced genes when the tubercle bacillus is subjected to hypoxia and nitrosative stress, conditions thought to promote latency. Induction of rv2623 also occurs when M. tuberculosis encounters conditions associated with growth arrest, such as the intracellular milieu of macrophages and in the lungs of mice with chronic tuberculosis. Therefore, we tested the hypothesis that Rv2623 regulates tuberculosis latency. We observed that an Rv2623-deficient mutant fails to establish chronic tuberculous infection in guinea pigs and mice, exhibiting a hypervirulence phenotype associated with increased bacterial burden and mortality. Consistent with this in vivo growth-regulatory role, constitutive overexpression of rv2623 attenuates mycobacterial growth in vitro. Biochemical analysis of purified Rv2623 suggested that this mycobacterial USP binds ATP, and the 2.9-A-resolution crystal structure revealed that Rv2623 engages ATP in a novel nucleotide-binding pocket. Structure-guided mutagenesis yielded Rv2623 mutants with reduced ATP-binding capacity. Analysis of mycobacteria overexpressing these mutants revealed that the in vitro growth-inhibitory property of Rv2623 correlates with its ability to bind ATP. Together, the results indicate that i M. tuberculosis Rv2623 regulates mycobacterial growth in vitro and in vivo, and ii Rv2623 is required for the entry of the tubercle bacillus into the chronic phase of infection in the host; in addition, iii Rv2623 binds ATP; and iv the growth-regulatory attribute of this USP is dependent on its ATP-binding activity. We propose that Rv2623 may function as an ATP-dependent signaling intermediate in a pathway that promotes persistent infection.

  6. Genetic Analysis of Mycobacterium avium Complex Strains Used for Producing Purified Protein Derivatives

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    Semret, Makeda; Bakker, Douwe; Smart, Nonie; Olsen, Ingrid; Haslov, Kaare; Behr, Marcel A.

    2006-01-01

    For over a century, purified protein derivatives (PPD) have been used to detect mycobacterial infections in humans and livestock. Among these, reagents to detect infections by Mycobacterium avium complex organisms have been produced, but the utility of these reagents has not been clearly established due in part to limited biologic and immunologic standardization. Because there is little information about the strains used to produce these reagents (avian PPD, intracellulare PPD, scrofulaceum PPD, and Johnin), we have performed genetic characterizations of strains used to produce these products. Sequence analysis of 16S rRNA and the hsp65 gene provided results concordant with species designations provided for M. avium, Mycobacterium intracellulare, and Mycobacterium scrofulaceum organisms. For M. avium strains, comparative genomic hybridization was performed on a whole-genome DNA microarray, revealing one novel 7.9-kilobase genomic deletion in certain Johnin-producing strains, in addition to genomic variability inherent to the particular M. avium subspecies. Our findings indicate that considerable genomic differences exist between organisms used for reagents and the infecting organism being studied. These results serve as a baseline for potency studies of different preparations and should aid in comparative studies of newly discovered antigens for the diagnosis of infection and disease by M. avium complex organisms. PMID:16960109

  7. Comparative analysis of mycobacterium and related actinomycetes yields insight into the evolution of mycobacterium tuberculosis pathogenesis

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    McGuire Abigail

    2012-03-01

    Full Text Available Abstract Background The sequence of the pathogen Mycobacterium tuberculosis (Mtb strain H37Rv has been available for over a decade, but the biology of the pathogen remains poorly understood. Genome sequences from other Mtb strains and closely related bacteria present an opportunity to apply the power of comparative genomics to understand the evolution of Mtb pathogenesis. We conducted a comparative analysis using 31 genomes from the Tuberculosis Database (TBDB.org, including 8 strains of Mtb and M. bovis, 11 additional Mycobacteria, 4 Corynebacteria, 2 Streptomyces, Rhodococcus jostii RHA1, Nocardia farcinia, Acidothermus cellulolyticus, Rhodobacter sphaeroides, Propionibacterium acnes, and Bifidobacterium longum. Results Our results highlight the functional importance of lipid metabolism and its regulation, and reveal variation between the evolutionary profiles of genes implicated in saturated and unsaturated fatty acid metabolism. It also suggests that DNA repair and molybdopterin cofactors are important in pathogenic Mycobacteria. By analyzing sequence conservation and gene expression data, we identify nearly 400 conserved noncoding regions. These include 37 predicted promoter regulatory motifs, of which 14 correspond to previously validated motifs, as well as 50 potential noncoding RNAs, of which we experimentally confirm the expression of four. Conclusions Our analysis of protein evolution highlights gene families that are associated with the adaptation of environmental Mycobacteria to obligate pathogenesis. These families include fatty acid metabolism, DNA repair, and molybdopterin biosynthesis. Our analysis reinforces recent findings suggesting that small noncoding RNAs are more common in Mycobacteria than previously expected. Our data provide a foundation for understanding the genome and biology of Mtb in a comparative context, and are available online and through TBDB.org.

  8. CUTANEOUS MYCOBACTERIUM MARINUM INFECTION DIAGNOSED BY PCR-RFLP ANALYSIS

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    LI Xiao-jie; WANG Hong-sheng; TAO Shi-qin; WU Qin-xue; LIU Wei-da

    2009-01-01

    Objective To identify Mycobacterium marinum (M. marinum) inducing misdiagnosis and treatment failure.Methods The lesional specimen of patient with cutaneous M. marinum were cultivated on Lwenstein-Jensen medium. The isolate was identified by biochemical tests and polymerase chain reaction (PCR)-restriction fragment length polymorphism (RFLP) analysis of the hsp65 gene.Results Smooth and non-pigmented colonies were noted after incubation at 32 ℃ for 2 weeks. The isolate was acid-fast bacilli and confirmed as M. marinum by biochemical tests and PCR-RFLP.Conclusion For a correct diagnosis of cutaneous M. marinum infection, it is crucial for clinicians to have a high index of suspicion, obtain the history of exposure and trauma and understand growth characteristics of the organism. Compared with conventional biochemical techniques, PCR-RFLP analysis is a more rapid, accurate and reliable method for mycobacterial identification to species level.

  9. AMPLIFIED FRAGMENT LENGTH POLYMORPHISM ANALYSIS OF MYCOBACTERIUM AVIUM COMPLEX ISOLATES RECOVERED FROM SOUTHERN CALIFORNIA

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    Fine-scale genotyping methods are necessary in order to identify possible sources of human exposure to opportunistic pathogens belonging to the Mycobacterium avium complex (MAC). In this study, amplified fragment length polymorphism (AFLP) analysis was evaluated for fingerprintin...

  10. Establishing Virulence Associated Polyphosphate Kinase 2 as a drug target for Mycobacterium tuberculosis.

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    Singh, Mamta; Tiwari, Prabhakar; Arora, Garima; Agarwal, Sakshi; Kidwai, Saqib; Singh, Ramandeep

    2016-01-01

    Inorganic polyphosphate (PolyP) plays an essential role in microbial stress adaptation, virulence and drug tolerance. The genome of Mycobacterium tuberculosis encodes for two polyphosphate kinases (PPK-1, Rv2984 and PPK-2, Rv3232c) and polyphosphatases (ppx-1, Rv0496 and ppx-2, Rv1026) for maintenance of intracellular PolyP levels. Microbial polyphosphate kinases constitute a molecular mechanism, whereby microorganisms utilize PolyP as phosphate donor for synthesis of ATP. In the present study we have constructed ppk-2 mutant strain of M. tuberculosis and demonstrate that PPK-2 enzyme contributes to its ability to cause disease in guinea pigs. We observed that ppk-2 mutant strain infected guinea pigs had significantly reduced bacterial loads and tissue pathology in comparison to wild type infected guinea pigs at later stages of infection. We also report that in comparison to the wild type strain, ppk-2 mutant strain was more tolerant to isoniazid and impaired for survival in THP-1 macrophages. In the present study we have standardized a luciferase based assay system to identify chemical scaffolds that are non-cytotoxic and inhibit M. tuberculosis PPK-2 enzyme. To the best of our knowledge this is the first study demonstrating feasibility of high throughput screening to obtain small molecule PPK-2 inhibitors. PMID:27279366

  11. IS6110限制性片段多态性分析标准方法的建立及其在结核分枝杆菌分子分型中的应用%Establishment and application of a standard IS6110-RFLP method in the study of molecular genotyping analysis on Mycobacterium tuberculosis

    Institute of Scientific and Technical Information of China (English)

    刘敬华; Kristin Kremer; Christine Pourcel; Arnout Mulder; 刘志广; 赵秀芹; 万康林

    2008-01-01

    Objective To develop a standardized IS6110-restriction fragment length polymorphism (RFLP) method, used for evaluating the capacity of genotyping. Methods IS6110-RFLP of 78 Mycobacterium (M.) tuberculosis strains were studied by bio-molecular techniques including DNA isolation, PCR, restriction endonuclease enzyme analysis, southern blotting, agarose gel electrophoresis,together with data analysis by software Gel-Pro analyzer 3.1 and BioNumerics (Version 5.0). Results IS6110-RFLP method was established and standardized successfully, including DNA isolation, PCR,restriction endonuclease enzyme analysis, southern blotting, agarose gel electrophoresis and usage of the analysis software with standard parameters. By this method, 78 M. tuberculosis isolates were classified into 75 genotypes which belonged to 11 different clusters. Of all the isolates, 66.7 % (52/78) belonged to a main cluster. Conclusion Standard IS6110-RFLP method was established successfully. This method had powerful capacity for genotyping and strain level identification and could be used for the surveillance on pathogens of M. tuberculosis in China.%目的 建立IS6110限制性片段多态性分析(IS6110-RFLP)标准方法 并评价该方法 的分型能力.方法 采用核酸提取、PCR、限制性内切酶分析、Southern杂交、琼脂糖凝胶电泳等技术,结合Gel-Pro analyzer 3.1和BioNumeries(Version 5.0)软件,对78株结核分枝杆菌插入序列IS6110-RFLP进行分析.结果 确定标准化的IS6110-RFLP技术,包括核酸提取、PCR、限制性内切酶分析、Southern杂交、琼脂糖凝胶电泳等实验步骤及标化参数的相关数据分析软件的使用;采用该技术,将78株结核分枝杆菌分为75个不同的基因型,分别归属于11个基因簇,其中有52株归属于同一个基因簇,占菌株总数的66.7%(52/78).结论 建立标准化的IS6110-RFLP技术方案,该方法 具有很强的基因分型和株水平鉴定能力,可用于结核病的病原学监测.

  12. CANDU safety analysis system establishment

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    Min, Byung Joo; Rhee, B. W.; Park, J. H.; Kim, H. T.; Choi, H. B.; Shim, J. I.; Yoon, C.; Yang, M. K

    2002-03-01

    To develop CANDU safety analysis system, methodology, and assessment technology, GAIs from CNSC and GSIs drived by IAEA are summarized. Furthermore, the following safety items are investigated in the present study. - It is intended to secure credibility of the void reactivity in the stage of nuclear design and analysis. The measurement data concerned with the void reactivity were reviewed and used to assess the physics code such as POWDERPUFS-V/RFSP, and the lattice code such as WIMS-AECL and MCNP-4B. - Reviewing the Final Safety Analysis Report for Wolsong-2/3/4 Units, the safety analysis methodology, classification for accident scenarios, safety analysis codes, their interface, etc. were examined. - The development of 3D CFD transient analysis model has been performed to predict local subcooling of the moderator in the vicinity of Calandria tubes in a CANDU-6 reactor in the case of Large LOCA transient. - The trip coverage analysis methodology based on CATHENA code is developed. The simulation of real plant transient showed good agreement. The trip coverage map was generated successfully for two typical depressurization and pressurization event. - The multi-dimensional analysis methodology for hydrogen distribution and hydrogen burning phenomena in PHWR containment is developed using GOTHIC code. The multi-dimensional analysis predicts the local hydrogen behaviour compared to the lumped parameter model.

  13. Mycobacterium tuberculosis H37Rv: In Silico Drug Targets Identification by Metabolic Pathways Analysis

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    Asad Amir

    2014-01-01

    Full Text Available Mycobacterium tuberculosis (Mtb is a pathogenic bacteria species in the genus Mycobacterium and the causative agent of most cases of tuberculosis. Tuberculosis (TB is the leading cause of death in the world from a bacterial infectious disease. This antibiotic resistance strain lead to development of the new antibiotics or drug molecules which can kill or suppress the growth of Mycobacterium tuberculosis. We have performed an in silico comparative analysis of metabolic pathways of the host Homo sapiens and the pathogen Mycobacterium tuberculosis (H37Rv. Novel efforts in developing drugs that target the intracellular metabolism of M. tuberculosis often focus on metabolic pathways that are specific to M. tuberculosis. We have identified five unique pathways for Mycobacterium tuberculosis having a number of 60 enzymes, which are nonhomologous to Homo sapiens protein sequences, and among them there were 55 enzymes, which are nonhomologous to Homo sapiens protein sequences. These enzymes were also found to be essential for survival of the Mycobacterium tuberculosis according to the DEG database. Further, the functional analysis using Uniprot showed involvement of all the unique enzymes in the different cellular components.

  14. Novel Species Including Mycobacterium fukienense sp. Is Found from Tuberc-ulosis Patients in Fujian Province, China, Using Phylogenetic Analysis of Mycobacterium chelonae/abscessus Complex

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    ZHANG Yuan Yuan; LI Yan Bing; HUANG Ming Xiang; ZHAO Xiu Qin; ZHANG Li Shui; LIU Wen En; WAN Kang Lin

    2013-01-01

    Objective To identify the novel species ‘Mycobacterium fukienense’ sp. nov of Mycobacterium chelonae/abscessus complex from tuberculosis patients in Fujian Province, China. Methods Five of 27 clinical Mycobacterium isolates (Cls) were previously identified as M. chelonae/abscessus complex by sequencing the hsp65, rpoB, 16S-23S rRNA internal transcribed spacer region (its), recA and sodA house-keeping genes commonly used to describe the molecular characteristics of Mycobacterium. Clinical Mycobacterium isolates were classified according to the gene sequence using a clustering analysis program. Sequence similarity within clusters and diversity between clusters were analyzed. Results The 5 isolates were identified with distinct sequences exhibiting 99.8% homology in the hsp65 gene. However, a complete lack of homology was observed among the sequences of the rpoB, 16S-23S rRNA internal transcribed spacer region (its), sodA, and recA genes as compared with the M. abscessus. Furthermore, no match for rpoB, sodA, and recA genes was identified among the published sequences. Conclusion The novel species, Mycobacterium fukienense, is identified from tuberculosis patients in Fujian Province, China, which does not belong to any existing subspecies of M. chelonea/abscessus complex.

  15. Comparative Genomics and Transcriptomic Analysis of Mycobacterium Kansasii

    KAUST Repository

    Alzahid, Yara

    2014-04-01

    The group of Mycobacteria is one of the most intensively studied bacterial taxa, as they cause the two historical and worldwide known diseases: leprosy and tuberculosis. Mycobacteria not identified as tuberculosis or leprosy complex, have been referred to by ‘environmental mycobacteria’ or ‘Nontuberculous mycobacteria (NTM). Mycobacterium kansasii (M. kansasii) is one of the most frequent NTM pathogens, as it causes pulmonary disease in immuno-competent patients and pulmonary, and disseminated disease in patients with various immuno-deficiencies. There have been five documented subtypes of this bacterium, by different molecular typing methods, showing that type I causes tuberculosis-like disease in healthy individuals, and type II in immune-compromised individuals. The remaining types are said to be environmental, thereby, not causing any diseases. The aim of this project was to conduct a comparative genomic study of M. kansasii types I-V and investigating the gene expression level of those types. From various comparative genomics analysis, provided genomics evidence on why M. kansasii type I is considered pathogenic, by focusing on three key elements that are involved in virulence of Mycobacteria: ESX secretion system, Phospholipase c (plcb) and Mammalian cell entry (Mce) operons. The results showed the lack of the espA operon in types II-V, which renders the ESX- 1 operon dysfunctional, as espA is one of the key factors that control this secretion system. However, gene expression analysis showed this operon to be deleted in types II, III and IV. Furthermore, plcB was found to be truncated in types III and IV. Analysis of Mce operons (1-4) show that mce-1 operon is duplicated, mce-2 is absent and mce-3 and mce-4 is present in one copy in M. kansasii types I-V. Gene expression profiles of type I-IV, showed that the secreted proteins of ESX-1 were slightly upregulated in types II-IV when compared to type I and the secreted forms of ESX-5 were highly down

  16. Cloning, expression, purification and bioinformatic analysis of 2-methylcitrate synthase from Mycobacterium tuberculosis

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    Kandasamy Eniyan; Urmi Bajpai

    2015-01-01

    Objective:To clone, express and purify2-methylcitrate synthase(Rv1131) gene of Mycobacterium tuberculosis(M. tuberculosis) and to study its structural characteristics using various bioinformatics tools.Methods:Rv1131 gene was amplified by polymerase chain reaction usingM. tuberculosisH37Rv genomicDNA and cloned into pGEM-T easy vector and sequenced. The gene was sub-cloned in pET28c vector, expressed inEscherichia coliBL21(E. coliBL21) (DE3) cells and the recombinant protein was identified byWestern blotting.The protein was purified usingNickel affinity chromatography and the structural characteristics like sub-cellular localization, presence of transmembrane helices and secondary structure of the protein were predicted by bioinformatics tools.Tertiary structure of the protein and phylogenetic analysis was also established byin silico analysis.Results:The expression of the recombinant protein (Rv1131) was confirmed by western blotting using anti-HIS antibodies and the protein was purified from the soluble fraction.In silicoanalysis showed that the protein contains no signal peptide and transmembrane helices.Active site prediction showed that the protein has histidine and aspartic acid residues at242,281 &332 positions respectively.Phylogenetic analysis showed 100% homology withmajor mycobacterial species.Secondary structure predicts2-methylcitrate synthase contain51.9% alpha-helix,8.7% extended strand and39.4% random coils.Tertiary structure of the protein was also established.Conclusions:The enzyme2-methylcitrate synthase from M. tuberculosisH37Rv has been successfully expressed and purified.The purified protein will further be utilized to develop assay methods for screening new inhibitors.

  17. Transcriptional analysis of Mycobacterium fortuitum cultures upon hydrogen peroxide treatment using the novel standard rrnA-P1

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    Rebollo María

    2008-06-01

    Full Text Available Abstract Background The ability of an intracellular pathogen to establish infection depends on the capacity of the organism to survive and replicate inside the host. Mycobacterium fortuitum is a bacteria that contains genes involved in the detoxification of the oxygen reactive species such as those produced by the host during the infection. In this work, we investigate the effects of hydrogen peroxide on the transcription and expression of these genes by developing a real time quantitative PCR technique (qRT-PCR using the ribosomal promoter region (rrnA-P1 as reference product for quantification of the mRNA levels. Results M. fortuitum cultures were treated with different hydrogen peroxide concentrations (0.02 to 20 mM during several periods of time (30 to 120 minutes. The activity of the enzymes KatGII and SodA, and the transcription of corresponding genes were evaluated. The transcriptional regulator furAII gene was also studied. The ribosomal promoter region rrnA-P1 was validated as referential product under the stress conditions checked by qRT-PCR. Minor changes were observed under the conditions tested except when bacteria were incubated in the presence of 20 mM hydrogen peroxide. Under those conditions, the levels of transcription of the three genes under study increased at 30 minutes of treatment. The viability of the bacteria was not influenced under the conditions tested. Conclusion In this work, we have quantified transcriptional responses to stress suggesting that, the opportunistic pathogen M. fortuitum is more resistant and differs in behaviour in the presence of hydrogen peroxide, when compared to the major pathogen Mycobacterium tuberculosis and the saprophyte Mycobacterium smegmatis. Besides, we demonstrate the mycobacterial non-coding region rrnA-P1 to be a suitable reference product in the analysis of qRT-PCR transcriptional data of M. fortuitum.

  18. Transcriptional analysis of Mycobacterium fortuitum cultures upon hydrogen peroxide treatment using the novel standard rrnA-P1

    Science.gov (United States)

    Núñez, María Carmen; Menéndez, María Carmen; Rebollo, María José; García, María J

    2008-01-01

    Background The ability of an intracellular pathogen to establish infection depends on the capacity of the organism to survive and replicate inside the host. Mycobacterium fortuitum is a bacteria that contains genes involved in the detoxification of the oxygen reactive species such as those produced by the host during the infection. In this work, we investigate the effects of hydrogen peroxide on the transcription and expression of these genes by developing a real time quantitative PCR technique (qRT-PCR) using the ribosomal promoter region (rrnA-P1) as reference product for quantification of the mRNA levels. Results M. fortuitum cultures were treated with different hydrogen peroxide concentrations (0.02 to 20 mM) during several periods of time (30 to 120 minutes). The activity of the enzymes KatGII and SodA, and the transcription of corresponding genes were evaluated. The transcriptional regulator furAII gene was also studied. The ribosomal promoter region rrnA-P1 was validated as referential product under the stress conditions checked by qRT-PCR. Minor changes were observed under the conditions tested except when bacteria were incubated in the presence of 20 mM hydrogen peroxide. Under those conditions, the levels of transcription of the three genes under study increased at 30 minutes of treatment. The viability of the bacteria was not influenced under the conditions tested. Conclusion In this work, we have quantified transcriptional responses to stress suggesting that, the opportunistic pathogen M. fortuitum is more resistant and differs in behaviour in the presence of hydrogen peroxide, when compared to the major pathogen Mycobacterium tuberculosis and the saprophyte Mycobacterium smegmatis. Besides, we demonstrate the mycobacterial non-coding region rrnA-P1 to be a suitable reference product in the analysis of qRT-PCR transcriptional data of M. fortuitum. PMID:18565220

  19. Subfractionation and analysis of the cell envelope (lipo)polysaccharides of Mycobacterium tuberculosis

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    Grzegorzewicz, Anna E.; Jackson, Mary

    2013-01-01

    The cell envelope of Mycobacterium tuberculosis, the causative agent of tuberculosis in humans, is the source of carbohydrates of exceptional structure which play essential roles in the physiology of the bacterium and in its interactions with the host during infection. Much of what is known about their biosynthesis was derived from the phenotypic analysis of knock-out or conditional knock-out mutants of Mycobacteria generated by random or specific insertional mutagenesis. Here, we describe th...

  20. Genomic Analysis of Pathogenicity Determinants in Mycobacterium kansasii Type I

    KAUST Repository

    Guan, Qingtian

    2016-05-01

    Mycobacteria, a genus within Actinobacteria Phylum, are well known for two pathogens that cause human diseases: leprosy and tuberculosis. Other than the obligate human mycobacteria, there is a group of bacteria that are present in the environment and occasionally cause diseases in immunocompromised persons: the non-tuberculosis mycobacteria (NTM). Mycobacterium kansasii, which was first discovered in the Kansas state, is the main etiologic agent responsible for lung infections caused by NTM and raises attention because of its co-infection with human immunodeficiency virus (HIV). Five subspecies of M. kansasii (Type I-V) were described and only M. kansasii Type I is pathogenic to humans. M. kansasii is a Gram-positive bacteria that has a unique cell wall and secretion system, which is essential for its pathogenicity. We undertook a comparative genomics and transcriptomic approach to identify components of M. kansasii Type I pathogenicity. Our previous study showed that espA (ESX-1 essential protein) operon, a major component of the secretion system, is exclusively present in M. kansasii Type I. The purpose of this study was to test the functional role of the espA operon in pathogenicity and identify other components that may also be involved in pathogenicity. This study provides a new molecular diagnostic method for M. kansasii Type I infection using PCR (Polymerase Chain Reaction) technique to target the espAoperon. With detailed manual curation of the comparative genomics datasets, we found several genes exclusively present in M. kansasii Type I including ppsA/ppsC and whiB6, that we believe are involved, or have an effect on ESX-mediated secretion system. We have also highlighted, in our study, the differences in genetic components coding for the cell membrane composition between the five subspecies of M. kansasii. These results shed light on genetic components that are responsible for pathogenicity determinants in Type I M. kansasii and may help to design better

  1. Proteomic analysis of streptomycin resistant and sensitive clinical isolates of Mycobacterium tuberculosis

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    Venkatesan Krishnamurthy

    2010-11-01

    Full Text Available Abstract Background Streptomycin (SM is a broad spectrum antibiotic and is an important component of any anti-tuberculosis therapy regimen. Several mechanisms have been proposed to explain the emergence of resistance but still our knowledge is inadequate. Proteins form a very complex network and drugs are countered by their modification/efflux or over expression/modification of targets. As proteins manifest most of the biological processes, these are attractive targets for developing drugs, immunodiagnostics or therapeutics. The aim of present study was to analyze and compare the protein profile of whole cell extracts from Mycobacterium tuberculosis clinical isolates susceptible and resistant to SM. Results Two-dimensional gel electrophoresis (2DE and matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF mass spectrometry was employed for analyzing the protein profiles. Homology and in silico characterization for identified proteins was assessed using BLAST, InterProScan and KEGG database searches. Computational studies on the possible interactions between SM and identified proteins were carried out by a battery of online servers and softwares, namely, CLUSTALW (KEGG, I-TASSER, VMD, PatchDock and FireDock. On comparing 2DE patterns, nine proteins were found consistently overexpressed in SM resistant isolates and were identified as Rv0350, Rv0440, Rv1240, Rv3075c, Rv2971, Rv3028c, Rv2145c, Rv2031c and Rv0569. In silico docking analysis showed significant interactions of SM with essential (Rv0350, Rv0440 and Rv2971 and non essential (Rv1240, Rv3075c and Rv2031c genes. Conclusions The computational results suggest high protein binding affinity of SM and suggested many possible interactions between identified proteins and the drug. Bioinformatic analysis proves attributive for analysis of diversity of proteins identified by whole proteome analysis. In-depth study of the these proteins will give an insight into probable sites of drug

  2. Rational engineering of a virulence gene from Mycobacterium tuberculosis facilitates proteomic analysis of a natural protein N-terminus.

    Science.gov (United States)

    Reyna, Cristal; Mba Medie, Felix; Champion, Matthew M; Champion, Patricia A

    2016-01-01

    Mass spectrometry (MS) for the detection of proteins is an indispensable tool for evaluating the biological processes of the proteome. Proteomics frequently requires proteolysis of proteins into peptide fragments. Proteins can be refractory to ideal proteolysis at the sequence level rendering them difficult to analyze by routine proteomics methods. EsxA (ESAT-6, Early Secreted Antigen, 6kDa) is a major virulence determinant of Mycobacterium tuberculosis, the cause of human tuberculosis. EsxA is routinely used to evaluate mycobacterial virulence in the laboratory and as a biomarker for tuberculosis in humans. The sequence of EsxA hinders deeper MS analysis beyond routine detection. Here we engineer the sequence of EsxA to add desirable tryptic properties aimed at improving complex MS analysis. We demonstrate that EsxA variants are amenable to MS analysis and remain functional in established in vitro and ex vivo assays of Esx-1-function. We provide the first demonstration of molecular engineering to specifically improve MS analysis of individual microbial proteins. PMID:27625110

  3. Rational engineering of a virulence gene from Mycobacterium tuberculosis facilitates proteomic analysis of a natural protein N-terminus

    Science.gov (United States)

    Reyna, Cristal; Mba Medie, Felix; Champion, Matthew M.; Champion, Patricia A.

    2016-01-01

    Mass spectrometry (MS) for the detection of proteins is an indispensable tool for evaluating the biological processes of the proteome. Proteomics frequently requires proteolysis of proteins into peptide fragments. Proteins can be refractory to ideal proteolysis at the sequence level rendering them difficult to analyze by routine proteomics methods. EsxA (ESAT-6, Early Secreted Antigen, 6kDa) is a major virulence determinant of Mycobacterium tuberculosis, the cause of human tuberculosis. EsxA is routinely used to evaluate mycobacterial virulence in the laboratory and as a biomarker for tuberculosis in humans. The sequence of EsxA hinders deeper MS analysis beyond routine detection. Here we engineer the sequence of EsxA to add desirable tryptic properties aimed at improving complex MS analysis. We demonstrate that EsxA variants are amenable to MS analysis and remain functional in established in vitro and ex vivo assays of Esx-1-function. We provide the first demonstration of molecular engineering to specifically improve MS analysis of individual microbial proteins. PMID:27625110

  4. Interferon-γ release assays for the diagnosis of latent Mycobacterium tuberculosis infection: a systematic review and meta-analysis

    DEFF Research Database (Denmark)

    Diel, R; Goletti, D; Ferrara, G;

    2011-01-01

    We conducted a systematic review and meta-analysis to compare the accuracy of the QuantiFERON-TB® Gold In-Tube (QFT-G-IT) and the T-SPOT®.TB assays with the tuberculin skin test (TST) for the diagnosis of latent Mycobacterium tuberculosis infection (LTBI). The Medline, Embase and Cochrane databas...

  5. Rapid detection and immune characterization of Mycobacterium abscessus infection in cystic fibrosis patients.

    Directory of Open Access Journals (Sweden)

    Mathis Steindor

    Full Text Available Cystic fibrosis patients are highly susceptible to infections with non-tuberculous mycobacteria. Especially Mycobacterium abscessus infections are common but reliable diagnosis is hampered by non-specific clinical symptoms and insensitive mycobacterial culture. In the present study we established novel methods for rapid detection and immune characterization of Mycobacterium abscessus infection in cystic fibrosis patients. We performed Mycobacterium abscessus specific DNA-strip- and quantitative PCR-based analyses of non-cultured sputum samples to detect and characterize Mycobacterium abscessus infections. Concomitantly in vitro T-cell reactivation with purified protein derivatives (PPDs from different mycobacterial species was used to determine Mycobacterium abscessus specific T-cell cytokine expression of infected cystic fibrosis patients. Four of 35 cystic fibrosis patients (11.4% were Mycobacterium abscessus culture positive and showed concordant DNA-strip-test results. Quantitative PCR revealed marked differences of mycobacterial burden between cystic fibrosis patients and during disease course. Tandem-repeat analysis classified distinct Mycobacterium abscessus strains of infected cystic fibrosis patients and excluded patient-to-patient transmission. Mycobacterium abscessus specific T-cells were detected in the blood of cystic fibrosis patients with confirmed chronic infection and a subgroup of patients without evidence of Mycobacterium abscessus infection. Comparison of cytokine expression and phenotypic markers revealed increased proportions of CD40L positive T-cells that lack Interleukin-2 expression as a marker for chronic Mycobacterium abscessus infections in cystic fibrosis patients. Direct sputum examination enabled rapid diagnosis and quantification of Mycobacterium abscessus in cystic fibrosis patients. T-cell in vitro reactivation and cytokine expression analyses may contribute to diagnosis of chronic Mycobacterium abscessus

  6. Failure of PCR-Based IS6110 Analysis To Detect Vertebral Spondylodiscitis Caused by Mycobacterium bovis

    OpenAIRE

    Steensels, Deborah; Fauville-Dufaux, Maryse; Boie, Johan; De Beenhouwer, Hans

    2013-01-01

    Mycobacterium bovis is responsible for a zoonosis originating in cattle. We report a case of a man with vertebral spondylodiscitis caused by Mycobacterium bovis. Diagnosis was complicated because of the lack of IS6110. These strains are rare, but microbiologists should be aware of their existence.

  7. Identification of Mycobacterium marinum 65 kD heat shock protein gene by polymerase chain reaction restriction analysis from lesions of swimming pool granuloma

    Institute of Scientific and Technical Information of China (English)

    2006-01-01

    Background Nontuberculous mycobacterium (NTM) had been reported to cause cutaneous infections which are difficult to interpret due to the variability of the clinical manifestations. Among NTM infections, Mycobacterium marinum (M. marinum) are mostly seen to cause skin infection. It is therefore important to establish a rapid approach for detection and identification of M. marinum from lesions of patients with suspected M. marinum infections. Methods Specimens were obtained from 5 patients with swimming pool granuloma. DNA was extracted and polymerase chain reaction (PCR) was performed. PCR products were digested with Hae III and BstE II, then analysed by pattern restriction analysis to detect heat shock protein (hsp) 65 kD gene. Results The 65 kD hsp gene was found in all specimens from patients with swimming pool granuloma. PCR restriction analysis (PRA) identified all 5 samples to be M. marinum infections, and the result was consistent with that of routine bacteriological identification. The lesions subsided or markedly improved upon treatment. Conclusions PRA is a sensitive, specific and rapid method in identification of mycobacteria. Application of this method will be helpful for early diagnosis of mycobacterial skin infections.

  8. Use of Mycobacterium smegmatis deficient in ADP-ribosyltransferase as surrogate for Mycobacterium tuberculosis in drug testing and mutation analysis.

    Science.gov (United States)

    Agrawal, Priyanka; Miryala, Sandeep; Varshney, Umesh

    2015-01-01

    Rifampicin (Rif) is a first line drug used for tuberculosis treatment. However, the emergence of drug resistant strains has necessitated synthesis and testing of newer analogs of Rif. Mycobacterium smegmatis is often used as a surrogate for M. tuberculosis. However, the presence of an ADP ribosyltransferase (Arr) in M. smegmatis inactivates Rif, rendering it impractical for screening of Rif analogs or other compounds when used in conjunction with them (Rif/Rif analogs). Rifampicin is also used in studying the role of various DNA repair enzymes by analyzing mutations in RpoB (a subunit of RNA polymerase) causing Rif resistance. These analyses use high concentrations of Rif when M. smegmatis is used as model. Here, we have generated M. smegmatis strains by deleting arr (Δarr). The M. smegmatis Δarr strains show minimum inhibitory concentration (MIC) for Rif which is similar to that for M. tuberculosis. The MICs for isoniazid, pyrazinamide, ethambutol, ciprofloxacin and streptomycin were essentially unaltered for M. smegmatis Δarr. The growth profiles and mutation spectrum of Δarr and, Δarr combined with ΔudgB (udgB encodes a DNA repair enzyme that excises uracil) strains were similar to their counterparts wild-type for arr. However, the mutation spectrum of ΔfpgΔarr strain differed somewhat from that of the Δfpg strain (fpg encodes a DNA repair enzyme that excises 8-oxo-G). Our studies suggest M. smegmatis Δarr strain as an ideal model system in drug testing and mutation spectrum determination in DNA repair studies.

  9. Establishment of fluorescence quantitative PCR assay in detection of Mycobacterium tuberculosis%结核分枝杆菌荧光定量PCR检测方法建立

    Institute of Scientific and Technical Information of China (English)

    徐菊玲; 徐伯赢; 周洪昌; 张慧; 段劲; 薛利军; 邵圣文

    2011-01-01

    目的 探讨结核分枝杆菌实时定量检测方法并进行评价.方法针对结核杆菌(Mycobacterium tuberculosis,Mtb)16S rDNA基因设计引物,应用SYBR Green I建立荧光定量PCR( FQ-PCR)反应体系;提取Mtb基因组DNA,PCR扩增16S rDNA片段,构建重组质粒pMD-TB16S;检测11份肺结核患者痰标本;以甲型链球菌、大肠杆菌等基因组DNA作对照,检验方法特异性,对同一份Mtb DNA模板进行批内和批间检测,计算变异系数(CV).结果靶向Mtb 16S rDNA基因引物能够特异扩增Mtb 16S rDNA基因,对照组细菌基因未见扩增,灵敏度为(Mtb基因组DNA) 1.2 pg/μL,即(38.9 +3.54)拷贝/μL的16S rDNA基因,批内和批间Ct值变异系数分别为0.27%和1.26%.结论以16S rDNA为靶基因的FQ-PCR技术,能够对Mtb进行快速、敏感而特异的定量检测.%Objective To establish and assess a rapid real-time quantifiable method for the detection of Mycobacterium tuberculosis (Mtb). Methods The specific primers targetting 16S rDNA gene in Mtb were designed and the fluorescence quantitative PCR(FQ-PCR) assay was performed with SYBR Green I. After extraction of Mtb genomic DNA.the 16S rDNA fragment was amplified by conventional PCR and used to construct recombinant pMD-TB16S plasmid. Then, the serial dilutions of pMD-TB16S plasmid were subjected to the quantitation standard curve in FQ-PCR assay. The sputum samples from 11 patients with pulmonary tuberculosis were detected by the same FQ-PCR, with various genomic DNAs of Streptococcus viridans,Staphylococcus epidermidis,Staphylococcus aureus,and Escherichia coli used as the negative control to confirm specificity of FQ-PCR assay. The stabiity was analyzed with inter and intra FQ-PCR test by using the same Mtb genomic DNA,and the coefficient of variation(CV) values of threshold cycle(Ct ) were calculated. Results The primers targetting Mtb 16S rDNA gene were specific to the amplification of 16S rDNA gene of Mtb but not other bacteria in control

  10. The PE-PPE Domain in Mycobacterium Reveals a Serine α/β Hydrolase Fold and Function: An In-Silico Analysis

    Science.gov (United States)

    Sultana, Rafiya; Tanneeru, Karunakar; Guruprasad, Lalitha

    2011-01-01

    The PE and PPE proteins first reported in the genome sequence of Mycobacterium tuberculosis strain H37Rv are now identified in all mycobacterial species. The PE-PPE domain (Pfam ID: PF08237) is a 225 amino acid residue conserved region located towards the C-terminus of some PE and PPE proteins and hypothetical proteins. Our in-silico sequence analysis revealed that this domain is present in all Mycobacteria, some Rhodococcus and Nocardia farcinica genomes. This domain comprises a pentapeptide sequence motif GxSxG/S at the N-terminus and conserved amino acid residues Ser, Asp and His that constitute a catalytic triad characteristic of lipase, esterase and cutinase activity. The fold prediction and comparative modeling of the 3-D structure of the PE-PPE domain revealed a “serine α/β hydrolase” structure with a central β-sheet flanked by α-helices on either side. The structure comprises a lid insertion with a closed structure conformation and has a solvent inaccessible active site. The oxyanion hole that stabilizes the negative charge on the tetrahedral intermediate has been identified. Our findings add to the growing list of serine hydrolases in mycobacterium, which are essential for the maintenance of their impermeable cell wall and virulence. These results provide the directions for the design of experiments to establish the function of PE and PPE proteins. PMID:21347309

  11. Typing of Human Mycobacterium avium Isolates in Italy by IS1245-Based Restriction Fragment Length Polymorphism Analysis

    Science.gov (United States)

    Lari, Nicoletta; Cavallini, Michela; Rindi, Laura; Iona, Elisabetta; Fattorini, Lanfranco; Garzelli, Carlo

    1998-01-01

    All but 2 of 63 Mycobacterium avium isolates from distinct geographic areas of Italy exhibited markedly polymorphic, multibanded IS1245 restriction fragment length polymorphism (RFLP) patterns; 2 isolates showed the low-number banding pattern typical of bird isolates. By computer analysis, 41 distinct IS1245 patterns and 10 clusters of essentially identical strains were detected; 40% of the 63 isolates showed genetic relatedness, suggesting the existence of a predominant AIDS-associated IS1245 RFLP pattern. PMID:9817900

  12. Phylogenetic analysis of vitamin B12-related metabolism in Mycobacterium tuberculosis

    Directory of Open Access Journals (Sweden)

    Douglas B. Young

    2015-03-01

    Full Text Available Comparison of genome sequences from clinical isolates of Mycobacterium tuberculosis with phylogenetically-related pathogens Mycobacterium marinum, Mycobacterium kansasii and Mycobacterium leprae reveals diversity amongst genes associated with vitamin B12-related metabolism. Diversity is generated by gene deletion events, differential acquisition of genes by horizontal transfer, and single nucleotide polymorphisms with predicted impact on protein function and transcriptional regulation. Differences in the B12 synthesis pathway, methionine biosynthesis, fatty acid catabolism, and DNA repair and replication are consistent with adaptations to different environmental niches and pathogenic lifestyles. While there is no evidence of further gene acquisition during expansion of the M. tuberculosis complex, the emergence of other forms of genetic diversity provides insights into continuing host-pathogen co-evolution and has the potential to identify novel targets for disease intervention.

  13. Sequence analysis corresponding to the PPE and PE proteins in Mycobacterium tuberculosis and other genomes

    Indian Academy of Sciences (India)

    Swathi Adindla; Lalitha Guruprasad

    2003-03-01

    Amino acid sequence analysis corresponding to the PPE proteins in H37Rv and CDC1551 strains of the Mycobacterium tuberculosis genomes resulted in the identification of a previously uncharacterized 225 amino acidresidue common region in 22 proteins. The pairwise sequence identities were as low as 18%. Conservation of amino acid residues was observed at fifteen positions that were distributed over the whole length of the region. The secondary structure corresponding to this region is predicted to be a mixture of -helices and -strands. Although the function is not known, proteins with this region specific to mycobacterial species may be associated with a common function. We further observed another group of 20 PPE proteins corresponding to the conserved C-terminal region comprising 44 amino acid residues with GFxGT and PxxPxxW sequence motifs. This region is preceded by a hydrophobic region, comprising 40–100 amino acid residues, that is flanked by charged amino acid residues. Identification of conserved regions described above may be useful to detect related proteins from other genomes and assist the design of suitable experiments to test their corresponding functions. Amino acid sequence analysis corresponding to the PE proteins resulted in the identification of tandem repeats comprising 41–43 amino acid residues in the C-terminal variable regions in two PE proteins (Rv0978 and Rv0980). These correspond to the AB repeats that were first identified in some proteins of the Methanosarcina mazei genome, and were demonstrated as surface antigens. We observed the AB repeats also in several other proteins of hitherto uncharacterized function in Archaea and Bacteria genomes. Some of these proteins are also associated with another repeat called the C-repeat or the PKD-domain comprising 85 amino acid residues. The secondary structure corresponding to the AB repeat is predicted mainly as 4 -strands. We suggest that proteins with AB repeats in Mycobacterium tuberculosis and

  14. Molecular Analysis of Mycobacterium avium Isolates by Using Pulsed-Field Gel Electrophoresis and PCR

    Science.gov (United States)

    Pestel-Caron, Martine; Graff, Gabriel; Berthelot, Gilles; Pons, Jean-Louis; Lemeland, Jean-François

    1999-01-01

    Genetic relationships among 46 isolates of Mycobacterium avium recovered from 37 patients in a 2,500-bed hospital from 1993 to 1998 were assessed by pulsed-field gel electrophoresis (PFGE) and PCR amplification of genomic sequences located between the repetitive elements IS1245 and IS1311. Each technique enabled the identification of 27 to 32 different patterns among the 46 isolates, confirming that the genetic heterogeneity of M. avium strains is high in a given community. Furthermore, this retrospective analysis of sporadic isolates allowed us (i) to suggest the existence of two remanent strains in our region, (ii) to raise the question of the possibility of nosocomial acquisition of M. avium strains, and (iii) to document laboratory contamination. The methods applied in the present study were found to be useful for the typing of M. avium isolates. In general, both methods yielded similar results for both related and unrelated isolates. However, the isolates in five of the six PCR clusters were distributed among two to three PFGE patterns, suggesting that this PCR-based method may have limitations for the analysis of strains with low insertion sequence copy numbers or for resolution of extended epidemiologic relationships. PMID:10405383

  15. Molecular Genetic Analysis of Multi-drug Resistance in Indian Isolates of Mycobacterium tuberculosis

    Directory of Open Access Journals (Sweden)

    Noman Siddiqi

    1998-09-01

    Full Text Available A total of 116 isolates from patients attending the out-patient department at the All India Institute of Medical Sciences, New Delhi and the New Delhi Tuberculosis Centre, New Delhi, India were collected. They were analyzed for resistance to drugs prescribed in the treatment for tuberculosis. The drug resistance was initially determined by microbiological techniques. The Bactec 460TB system was employed to determine the type and level of resistance in each isolate. The isolates were further characterized at molecular level. The multi-drug loci corresponding to rpo b, gyr A, kat G were studied for mutation(s by the polymerase chain reaction-single strand conformational polymorphism (PCR-SSCP technique. The SSCP positive samples were sequenced to characterize the mutations in rpo b, and gyr A loci. While previously reported mutations in the gyr A and rpo b loci were found to be present, several novel mutations were also scored in the rpo b locus. Interestingly, analysis of the gyr A locus showed the presence of point mutation(s that could not be detected by PCR-SSCP. Furthermore, rifampicin resistance was found to be an important marker for checking multi-drug resistance (MDR in clinical isolates of Mycobacterium tuberculosis. This is the first report on molecular genetic analysis of MDR tuberculosis one from India, highlights the increasing incidence of MDR in the Indian isolates of M. tuberculosis.

  16. Proteome analysis of the Mycobacterium tuberculosis Beijing B0/W148 cluster.

    Science.gov (United States)

    Bespyatykh, Julia; Shitikov, Egor; Butenko, Ivan; Altukhov, Ilya; Alexeev, Dmitry; Mokrousov, Igor; Dogonadze, Marine; Zhuravlev, Viacheslav; Yablonsky, Peter; Ilina, Elena; Govorun, Vadim

    2016-01-01

    Beijing B0/W148, a "successful" clone of Mycobacterium tuberculosis, is widespread in the Russian Federation and some countries of the former Soviet Union. Here, we used label-free gel-LC-MS/MS shotgun proteomics to discover features of Beijing B0/W148 strains that could explain their success. Qualitative and quantitative proteome analyses of Beijing B0/W148 strains allowed us to identify 1,868 proteins, including 266 that were differentially abundant compared with the control strain H37Rv. To predict the biological effects of the observed differences in protein abundances, we performed Gene Ontology analysis together with analysis of protein-DNA interactions using a gene regulatory network. Our results demonstrate that Beijing B0/W148 strains have increased levels of enzymes responsible for long-chain fatty acid biosynthesis, along with a coincident decrease in the abundance of proteins responsible for their degradation. Together with high levels of HsaA (Rv3570c) protein, involved in steroid degradation, these findings provide a possible explanation for the increased transmissibility of Beijing B0/W148 strains and their survival in host macrophages. Among other, we confirmed a very low level of the SseA (Rv3283) protein in Beijing B0/W148 characteristic for all «modern» Beijing strains, which could lead to increased DNA oxidative damage, accumulation of mutations, and potentially facilitate the development of drug resistance. PMID:27356881

  17. Functional analysis of TPM domain containing Rv2345 of Mycobacterium tuberculosis identifies its phosphatase activity.

    Science.gov (United States)

    Sinha, Avni; Eniyan, Kandasamy; Sinha, Swati; Lynn, Andrew Michael; Bajpai, Urmi

    2015-07-01

    Mycobacterium tuberculosis (Mtb) is the causal agent of tuberculosis, the second largest infectious disease. With the rise of multi-drug resistant strains of M. tuberculosis, serious challenge lies ahead of us in treating the disease. The availability of complete genome sequence of Mtb has improved the scope for identifying new proteins that would not only further our understanding of biology of the organism but could also serve to discover new drug targets. In this study, Rv2345, a hypothetical membrane protein of M. tuberculosis H37Rv, which is reported to be a putative ortholog of ZipA cell division protein has been assigned function through functional annotation using bioinformatics tools followed by experimental validation. Sequence analysis showed Rv2345 to have a TPM domain at its N-terminal region and predicted it to have phosphatase activity. The TPM domain containing region of Rv2345 was cloned and expressed using pET28a vector in Escherichia coli and purified by Nickel affinity chromatography. The purified TPM domain was tested in vitro and our results confirmed it to have phosphatase activity. The enzyme activity was first checked and optimized with pNPP as substrate, followed by using ATP, which was also found to be used as substrate by the purified protein. Hence sequence analysis followed by in vitro studies characterizes TPM domain of Rv2345 to contain phosphatase activity.

  18. Heterogeneity among Mycobacterium ulcerans from French Guiana revealed by multilocus variable number tandem repeat analysis (MLVA).

    Science.gov (United States)

    Reynaud, Yann; Millet, Julie; Couvin, David; Rastogi, Nalin; Brown, Christopher; Couppié, Pierre; Legrand, Eric

    2015-01-01

    Buruli ulcer is an emerging and neglected tropical disease caused by Mycobacterium ulcerans. Few cases have been reported so far in the Americas. With 250 cases reported since 1969, French Guiana is the only Buruli ulcer endemic area in the continent. Thus far, no genetic diversity studies of strains of M. ulcerans from French Guiana have been reported. Our goal in the present study was to examine the genetic diversity of M. ulcerans strains in this region by using the Multilocus Variable Number Tandem Repeat Analysis (MLVA) approach. A total of 23 DNA samples were purified from ulcer biopsies or derived from pure cultures. MVLA was used in the study of six previously-described Variable Number of Tandem Repeat (VNTR) markers. A total of three allelic combinations were characterized in our study: genotype I which has been described previously, genotype III which is very similar to genotype I, and genotype II which has distinctly different characteristics in comparison with the other two genotypes. This high degree of genetic diversity appears to be uncommon for M. ulcerans. Further research based on complete genome sequencing of strains belonging to genotypes I and II is in progress and should lead soon to a better understanding of genetic specificities of M. ulcerans strains from French Guiana.

  19. Mycobacterium tuberculosis complex differentiation using gyrB-restriction fragment length polymorphism analysis

    Directory of Open Access Journals (Sweden)

    Erica Chimara

    2004-11-01

    Full Text Available Mycobacterium tuberculosis complex (MTBC members are causative agents of human and animal tuberculosis. Differentiation of MTBC members is required for appropriate treatment of individual patients and for epidemiological purposes. Strains from six MTBC species - M. tuberculosis, M. bovis subsp. bovis, M. bovis BCG, M. africanum, M. pinnipedii, and "M. canetti" - were studied using gyrB-restriction fragment length polymorphism (gyrB-RFLP analysis. A table was elaborated, based on observed restriction patterns and published gyrB sequences. To evaluate applicability of gyrB-RFLP at Instituto Adolfo Lutz, São Paulo, Mycobacterial Reference Laboratory, 311 MTBC clinical isolates, previously identified using traditional methods as M. tuberculosis (306, M. bovis (3, and M. bovis BCG (2, were analyzed by gyrB-RFLP. All isolates were correctly identified by the molecular method, but no distinction between M. bovis and M. bovis BCG was obtained. Differentiation of M. tuberculosis and M. bovis is of utmost importance, because they require different treatment schedules. In conclusion, gyrB-RFLP is accurate and easy-to-perform, with potential to reduce time needed for conventional differentiation methods. However, application for epidemiological studies remains limited, because it cannot differentiate M. tuberculosis from M. africanum subtype II, and "M. canetti", M. africanum subtype I from M. pinnipedii, and. M. bovis from M. bovis BCG.

  20. Analysis on Factors Affecting Seedling Establishment in Rice

    Institute of Scientific and Technical Information of China (English)

    LUO Ju; TANG Shao-qing; HU Pei-song; Aleman LOUIS; JIAO Gui-ai; TANG Jian

    2007-01-01

    Elongations of coleoptile and mesocotyl are related directly to rice seedling establishment in soil and height of plant is related to lodging in rice production. Twelve typical rice cultivars with different lengths of coleoptile and mesocotyl (long, medium and short) were selected by screening the lengths of coleoptile and mesocotyl in 1500 accessions. The seedling establishments of these typical cultivars were compared under the combinations of different sowing depths and flooding durations, and two semi-dwarf varieties (G140, Zhong 96-21) with good seedling establishments and optimum mesocotyl lengths were found. The length of mesocotyl was completely fitted negative binomial distribution and the length of coleoptile was nearly fitted Iognormal distribution.Analysis of the relationships among mesocotyl, coleoptile, seeding depth, flooding duration, and their interactions to seedling establishment percentage showed that there existed significant relations among mesocotyl, coleoptile, mesocotyl × coleoptile,seeding depth, flooding duration and mesocotyl × sowing depth in the experiment for seedling establishment.

  1. Phosphoproteomics analysis of a clinical Mycobacterium tuberculosis Beijing isolate: Expanding the mycobacterial phosphoproteome catalogue

    Directory of Open Access Journals (Sweden)

    Suereta eFortuin

    2015-02-01

    Full Text Available Reversible protein phosphorylation, regulated by protein kinases and phosphatases, mediates a switch between protein activity and cellular pathways that contribute to a large number of cellular processes. The Mycobacterium tuberculosis genome encodes 11 Serine/Threonine kinases (STPKs which show close homology to eukaryotic kinases. This study aimed to elucidate the phosphoproteomic landscape of a clinical isolate of M. tuberculosis. We performed a high throughput mass spectrometric analysis of proteins extracted from an early-logarithmic phase culture. Whole cell lysate proteins were processed using the filter-aided sample preparation method, followed by phosphopeptide enrichment of tryptic peptides by strong cation exchange (SCX and Titanium dioxide (TiO2 chromatography. The MaxQuant quantitative proteomics software package was used for protein identification. Our analysis identified 414 serine/threonine/tyrosine phosphorylated sites, with a distribution of S/T/Y sites; 38% on serine, 59% on threonine and 3% on tyrosine; present on 303 unique peptides mapping to 214 M. tuberculosis proteins. Only forty five of the S/T/Y phosphorylated proteins identified in our study had been previously described in the laboratory strain H37Rv, confirming previous reports. The remaining 169 phosphorylated proteins were newly identified in this clinical M. tuberculosis Beijing strain. We identified 5 novel tyrosine phosphorylated proteins. These findings not only expand upon our current understanding of the protein phosphorylation network in clinical M. tuberculosis but the data set also further extends and complements previous knowledge regarding phosphorylated peptides and phosphorylation sites in M. tuberculosis.

  2. Comparative genomics analysis of Mycobacterium ulcerans for the identification of putative essential genes and therapeutic candidates.

    Directory of Open Access Journals (Sweden)

    Azeem Mehmood Butt

    Full Text Available Mycobacterium ulcerans, the causative agent of Buruli ulcer, is the third most common mycobacterial disease after tuberculosis and leprosy. The present treatment options are limited and emergence of treatment resistant isolates represents a serious concern and a need for better therapeutics. Conventional drug discovery methods are time consuming and labor-intensive. Unfortunately, the slow growing nature of M. ulcerans in experimental conditions is also a barrier for drug discovery and development. In contrast, recent advancements in complete genome sequencing, in combination with cheminformatics and computational biology, represent an attractive alternative approach for the identification of therapeutic candidates worthy of experimental research. A computational, comparative genomics workflow was defined for the identification of novel therapeutic candidates against M. ulcerans, with the aim that a selected target should be essential to the pathogen, and have no homology in the human host. Initially, a total of 424 genes were predicted as essential from the M. ulcerans genome, via homology searching of essential genome content from 20 different bacteria. Metabolic pathway analysis showed that the most essential genes are associated with carbohydrate and amino acid metabolism. Among these, 236 proteins were identified as non-host and essential, and could serve as potential drug and vaccine candidates. Several drug target prioritization parameters including druggability were also calculated. Enzymes from several pathways are discussed as potential drug targets, including those from cell wall synthesis, thiamine biosynthesis, protein biosynthesis, and histidine biosynthesis. It is expected that our data will facilitate selection of M. ulcerans proteins for successful entry into drug design pipelines.

  3. Spoligotyping and variable number tandem repeat analysis of Mycobacterium bovis isolates from cattle in Brazil

    Directory of Open Access Journals (Sweden)

    Patrícia Martins Parreiras

    2012-02-01

    Full Text Available We performed spoligotyping and 12-mycobacterial interspersed repetitive unit-variable number tandem repeats (MIRU-VNTRs typing to characterise Mycobacterium bovis isolates collected from tissue samples of bovines with lesions suggestive for tuberculosis during slaughter inspection procedures in abattoirs in Brazil. High-quality genotypes were obtained with both procedures for 61 isolates that were obtained from 185 bovine tissue samples and all of these isolates were identified as M. bovis by conventional identification procedures. On the basis of the spoligotyping, 53 isolates were grouped into nine clusters and the remaining eight isolates were unique types, resulting in 17 spoligotypes. The majority of the Brazilian M. bovis isolates displayed spoligotype patterns that have been previously observed in strains isolated from cattle in other countries. MIRU-VNTR typing produced 16 distinct genotypes, with 53 isolates forming eight of the groups, and individual isolates with unique VNTR profiles forming the remaining eight groups. The allelic diversity of each VNTR locus was calculated and only two of the 12-MIRU-VNTR loci presented scores with either a moderate (0.4, MIRU16 or high (0.6, MIRU26 discriminatory index (h. Both typing methods produced similar discriminatory indexes (spoligotyping h = 0.85; MIRU-VNTR h = 0.86 and the combination of the two methods increased the h value to 0.94, resulting in 29 distinct patterns. These results confirm that spoligotyping and VNTR analysis are valuable tools for studying the molecular epidemiology of M. bovis infections in Brazil.

  4. Amplified fragment length polymorphism analysis of human clinical isolates of Mycobacterium haemophilum from different continents

    NARCIS (Netherlands)

    L.E.S. Bruijnesteijn van Coppenraet; P.H.M. Savelkoul; N. Buffing; M.W. van der Bijl; J. Woudenberg; J.A. Lindeboom; T.E. Kiehn; F. Haverkort; Z. Samra; E.J. Kuijper

    2009-01-01

    The role of the species Mycobacterium haemophilum as a pathogenic non-tuberculous microorganism is becoming better defined with the use of specific detection methods. However, epidemiological investigations of this species are still scarce. We analysed the genetic diversity of M. haemophilum by ampl

  5. Spatial analysis of factors implicated in Mycobacterium ulcerans infection in Ghana

    NARCIS (Netherlands)

    Duker, A.A.

    2005-01-01

    Buruliulcer (BU), the common terminology for the disease caused by Mycobacteriumulcerans (MU) infection manifests as disfiguring skin ulceration which is difficult to treat. In its advanced st

  6. Establishment and validation of whole-cell based fluorescence assays to identify anti-mycobacterial compounds using the Acanthamoeba castellanii-Mycobacterium marinum host-pathogen system.

    Directory of Open Access Journals (Sweden)

    Sébastien Kicka

    Full Text Available Tuberculosis is considered to be one of the world's deadliest disease with 2 million deaths each year. The need for new antitubercular drugs is further exacerbated by the emergence of drug-resistance strains. Despite multiple recent efforts, the majority of the hits discovered by traditional target-based screening showed low efficiency in vivo. Therefore, there is heightened demand for whole-cell based approaches directly using host-pathogen systems. The phenotypic host-pathogen assay described here is based on the monitoring of GFP-expressing Mycobacterium marinum during infection of the amoeba Acanthamoeba castellanii. The assay showed straight-forward medium-throughput scalability, robustness and ease of manipulation, demonstrating its qualities as an efficient compound screening system. Validation with a series of known antitubercular compounds highlighted the advantages of the assay in comparison to previously published macrophage-Mycobacterium tuberculosis-based screening systems. Combination with secondary growth assays based on either GFP-expressing D. discoideum or M. marinum allowed us to further fine-tune compound characterization by distinguishing and quantifying growth inhibition, cytotoxic properties and antibiotic activities of the compounds. The simple and relatively low cost system described here is most suitable to detect anti-infective compounds, whether they present antibiotic activities or not, in which case they might exert anti-virulence or host defense boosting activities, both of which are largely overlooked by classical screening approaches.

  7. Development and analysis of an in vivo-compatible metabolic network of Mycobacterium tuberculosis

    Directory of Open Access Journals (Sweden)

    Reifman Jaques

    2010-11-01

    Full Text Available Abstract Background During infection, Mycobacterium tuberculosis confronts a generally hostile and nutrient-poor in vivo host environment. Existing models and analyses of M. tuberculosis metabolic networks are able to reproduce experimentally measured cellular growth rates and identify genes required for growth in a range of different in vitro media. However, these models, under in vitro conditions, do not provide an adequate description of the metabolic processes required by the pathogen to infect and persist in a host. Results To better account for the metabolic activity of M. tuberculosis in the host environment, we developed a set of procedures to systematically modify an existing in vitro metabolic network by enhancing the agreement between calculated and in vivo-measured gene essentiality data. After our modifications, the new in vivo network contained 663 genes, 838 metabolites, and 1,049 reactions and had a significantly increased sensitivity (0.81 in predicted gene essentiality than the in vitro network (0.31. We verified the modifications generated from the purely computational analysis through a review of the literature and found, for example, that, as the analysis suggested, lipids are used as the main source for carbon metabolism and oxygen must be available for the pathogen under in vivo conditions. Moreover, we used the developed in vivo network to predict the effects of double-gene deletions on M. tuberculosis growth in the host environment, explore metabolic adaptations to life in an acidic environment, highlight the importance of different enzymes in the tricarboxylic acid-cycle under different limiting nutrient conditions, investigate the effects of inhibiting multiple reactions, and look at the importance of both aerobic and anaerobic cellular respiration during infection. Conclusions The network modifications we implemented suggest a distinctive set of metabolic conditions and requirements faced by M. tuberculosis during

  8. Analysis of Mycobacterium tuberculosis Genotypic Lineage Distribution in Chile and Neighboring Countries.

    Science.gov (United States)

    Lagos, Jaime; Couvin, David; Arata, Loredana; Tognarelli, Javier; Aguayo, Carolina; Leiva, Tamara; Arias, Fabiola; Hormazabal, Juan Carlos; Rastogi, Nalin; Fernández, Jorge

    2016-01-01

    Tuberculosis (TB), caused by the pathogen Mycobacterium tuberculosis (MTB), remains a disease of high importance to global public health. Studies into the population structure of MTB have become vital to monitoring possible outbreaks and also to develop strategies regarding disease control. Although Chile has a low incidence of MTB, the current rates of migration have the potential to change this scenario. We collected and analyzed a total of 458 M. tuberculosis isolates (1 isolate per patient) originating from all 15 regions of Chile. The isolates were genotyped using the spoligotyping method and the data obtained were analyzed and compared with the SITVIT2 database. A total of 169 different patterns were identified, of which, 119 patterns (408 strains) corresponded to Spoligotype International Types (SITs) and 50 patterns corresponded to orphan strains. The most abundantly represented SITs/lineages were: SIT53/T1 (11.57%), SIT33/LAM3 (9.6%), SIT42/LAM9 (9.39%), SIT50/H3 (5.9%), SIT37/T3 (5%); analysis of the spoligotyping minimum spanning tree as well as spoligoforest were suggestive of a recent expansion of SIT42, SIT50 and SIT37; all of which potentially evolved from SIT53. The most abundantly represented lineages were LAM (40.6%), T (34.1%) and Haarlem (13.5%). LAM was more prevalent in the Santiago (43.6%) and Concepción (44.1%) isolates, rather than the Iquique (29.4%) strains. The proportion of X lineage was appreciably higher in Iquique and Concepción (11.7% in both) as compared to Santiago (1.6%). Global analysis of MTB lineage distribution in Chile versus neighboring countries showed that evolutionary recent lineages (LAM, T and Haarlem) accounted together for 88.2% of isolates in Chile, a pattern which mirrored MTB lineage distribution in neighboring countries (n = 7378 isolates recorded in SITVIT2 database for Peru, Brazil, Paraguay, and Argentina; and published studies), highlighting epidemiological advantage of Euro-American lineages in this region

  9. Analysis of Mycobacterium tuberculosis Genotypic Lineage Distribution in Chile and Neighboring Countries

    Science.gov (United States)

    Lagos, Jaime; Couvin, David; Arata, Loredana; Tognarelli, Javier; Aguayo, Carolina; Leiva, Tamara; Arias, Fabiola; Hormazabal, Juan Carlos; Rastogi, Nalin; Fernández, Jorge

    2016-01-01

    Tuberculosis (TB), caused by the pathogen Mycobacterium tuberculosis (MTB), remains a disease of high importance to global public health. Studies into the population structure of MTB have become vital to monitoring possible outbreaks and also to develop strategies regarding disease control. Although Chile has a low incidence of MTB, the current rates of migration have the potential to change this scenario. We collected and analyzed a total of 458 M. tuberculosis isolates (1 isolate per patient) originating from all 15 regions of Chile. The isolates were genotyped using the spoligotyping method and the data obtained were analyzed and compared with the SITVIT2 database. A total of 169 different patterns were identified, of which, 119 patterns (408 strains) corresponded to Spoligotype International Types (SITs) and 50 patterns corresponded to orphan strains. The most abundantly represented SITs/lineages were: SIT53/T1 (11.57%), SIT33/LAM3 (9.6%), SIT42/LAM9 (9.39%), SIT50/H3 (5.9%), SIT37/T3 (5%); analysis of the spoligotyping minimum spanning tree as well as spoligoforest were suggestive of a recent expansion of SIT42, SIT50 and SIT37; all of which potentially evolved from SIT53. The most abundantly represented lineages were LAM (40.6%), T (34.1%) and Haarlem (13.5%). LAM was more prevalent in the Santiago (43.6%) and Concepción (44.1%) isolates, rather than the Iquique (29.4%) strains. The proportion of X lineage was appreciably higher in Iquique and Concepción (11.7% in both) as compared to Santiago (1.6%). Global analysis of MTB lineage distribution in Chile versus neighboring countries showed that evolutionary recent lineages (LAM, T and Haarlem) accounted together for 88.2% of isolates in Chile, a pattern which mirrored MTB lineage distribution in neighboring countries (n = 7378 isolates recorded in SITVIT2 database for Peru, Brazil, Paraguay, and Argentina; and published studies), highlighting epidemiological advantage of Euro-American lineages in this region

  10. Risk Analysis as Regulatory Science: Toward The Establishment of Standards.

    Science.gov (United States)

    Murakami, Michio

    2016-09-01

    Understanding how to establish standards is essential for risk communication and also provides perspectives for further study. In this paper, the concept of risk analysis as regulatory science for the establishment of standards is demonstrated through examples of standards for evacuation and provisional regulation values in foods and drinking water. Moreover, academic needs for further studies related to standards are extracted. The concepts of the traditional 'Standard I', which has a paternalistic orientation, and 'Standard II', established through stakeholder consensus, are then systemized by introducing the current status of the new standards-related movement that developed after the Fukushima nuclear power plant accident, and the perspectives of the standards are discussed. Preparation of standards on the basis of stakeholder consensus through intensive risk dialogue before a potential nuclear power plant accident is suggested to be a promising approach to ensure a safe society and enhance subjective well-being.

  11. Molecular Analysis of Mycobacterium avium Isolates by Using Pulsed-Field Gel Electrophoresis and PCR

    OpenAIRE

    Pestel-Caron, Martine; Graff, Gabriel; Berthelot, Gilles; Pons, Jean-Louis; Lemeland, Jean-François

    1999-01-01

    Genetic relationships among 46 isolates of Mycobacterium avium recovered from 37 patients in a 2,500-bed hospital from 1993 to 1998 were assessed by pulsed-field gel electrophoresis (PFGE) and PCR amplification of genomic sequences located between the repetitive elements IS1245 and IS1311. Each technique enabled the identification of 27 to 32 different patterns among the 46 isolates, confirming that the genetic heterogeneity of M. avium strains is high in a given community. Furthermore, this ...

  12. Molecular Analysis of Cross-Resistance to Capreomycin, Kanamycin, Amikacin, and Viomycin in Mycobacterium tuberculosis

    OpenAIRE

    Maus, Courtney E.; Plikaytis, Bonnie B.; Shinnick, Thomas M

    2005-01-01

    Capreomycin, kanamycin, amikacin, and viomycin are drugs that are used to treat multidrug-resistant tuberculosis. Each inhibits translation, and cross-resistance to them is a concern during therapy. A recent study revealed that mutation of the tlyA gene, encoding a putative rRNA methyltransferase, confers capreomycin and viomycin resistance in Mycobacterium tuberculosis bacteria. Mutations in the 16S rRNA gene (rrs) have been associated with resistance to each of the drugs; however, reports o...

  13. Molecular analysis of Mycobacterium isolates from extrapulmonary specimens obtained from patients in Mexico

    Directory of Open Access Journals (Sweden)

    Portillo-Gómez Leopoldo

    2009-03-01

    Full Text Available Abstract Background Little information is available on the molecular epidemiology in Mexico of Mycobacterium species infecting extrapulmonary sites in humans. This study used molecular methods to determine the Mycobacterium species present in tissues and body fluids in specimens obtained from patients in Mexico with extrapulmonary disease. Methods Bacterial or tissue specimens from patients with clinical or histological diagnosis of extrapulmonary tuberculosis were studied. DNA extracts from 30 bacterial cultures grown in Löwenstein Jensen medium and 42 paraffin-embedded tissues were prepared. Bacteria were cultured from urine, cerebrospinal fluid, pericardial fluid, gastric aspirate, or synovial fluid samples. Tissues samples were from lymph nodes, skin, brain, vagina, and peritoneum. The DNA extracts were analyzed by PCR and by line probe assay (INNO-LiPA MYCOBACTERIA v2. Innogenetics NV, Gent, Belgium in order to identify the Mycobacterium species present. DNA samples positive for M. tuberculosis complex were further analyzed by PCR and line probe assay (INNO-LiPA Rif.TB, Innogenetics NV, Gent, Belgium to detect mutations in the rpoB gene associated with rifampicin resistance. Results Of the 72 DNA extracts, 26 (36.1% and 23 (31.9% tested positive for Mycobacterium species by PCR or line probe assay, respectively. In tissues, M. tuberculosis complex and M. genus were found in lymph nodes, and M. genus was found in brain and vagina specimens. In body fluids, M. tuberculosis complex was found in synovial fluid. M. gordonae, M. smegmatis, M. kansasii, M. genus, M. fortuitum/M. peregrinum complex and M. tuberculosis complex were found in urine. M. chelonae/M. abscessus was found in pericardial fluid and M. kansasii was found in gastric aspirate. Two of M. tuberculosis complex isolates were also PCR and LiPA positive for the rpoB gene. These two isolates were from lymph nodes and were sensitive to rifampicin. Conclusion 1 We describe the

  14. Usefulness of three-channel multiplex real-time PCR and melting curve analysis for simultaneous detection and identification of the Mycobacterium tuberculosis complex and nontuberculous mycobacteria.

    Science.gov (United States)

    Hong, Yun Ji; Chung, Young Hoon; Kim, Taek Soo; Song, Sang Hoon; Park, Kyoung Un; Yim, Jae Joon; Song, Junghan; Lee, Jae Ho; Kim, Eui Chong

    2011-11-01

    We attempted to determine the benefits of three-channel multiplex real-time PCR and melting curve analysis not only in detecting and distinguishing between nontuberculous mycobacteria (NTM) and the Mycobacterium tuberculosis complex but also in identifying NTM to the species level.

  15. [Frontier of mycobacterium research--host vs. mycobacterium].

    Science.gov (United States)

    Okada, Masaji; Shirakawa, Taro

    2005-09-01

    of M. tuberculosis and the development of novel therapy. Dr. Chiyoji Abe (Nippon Becton Dickinson Co.) reviewed the molecular basis of the resistance to anti-tuberculosis drugs. Most cases of resistance are related to simple nucleotide substitutions rather than to acquisition of new elements. Dr. Kiyoshi Takeda (Kyushu University) showed interesting finding. He analyzed whether Toll-like receptor (TLR)-mediated activation of innate immunity in host defense against mycobacterial infection. MyD88/TRIF double defi-indicating that innate immunity is involved in anti-mycobacterial infection. (1) SNP (single nucleotide polymorphism) analysis in association with Mycobacterium tuberculosis: Taro SHIRAKAWA (Department of Health Promotion & Human Behavior, Kyoto University Medical School, and RIKEN SRC Center) Candidate gene approach was made on 18 SNPs in 11 genes in association with M. tuberculosis. Patients with multi-drug resistance against M. tuberculosis are also subjected. SNPs in NRAMP1 gene were associated with the disease, and drug resistance, its mechanisms remain unknown. (2) Search for genes susceptible to pulmonary Mycobacterium avium complex infection: Naoto KEICHO (Department of Respiratory Diseases, Research Institute, International Medical Center of Japan) Interaction among pathogens and host factors is important for development of infectious diseases. We are trying to identify host genetic factors involved in susceptibility to nonimmunocompromized pulmonary Mycobacterium avium complex (MAC) infection by candidate gene approach and genome-wide approach. Elucidation of functional significance of susceptibility gene polymorphisms will lead to a new strategy for control and prevention of the disease. (3) T cell immunity against Tuberculosis in host and the establishment of novel vaccine: Masaji OKADA (Clinical Research Center, National Hospital Organization Kinki-Chuo Chest Medical Center) T cell (CTL, Th1) immunity including granulysin play an important role in

  16. The epidemiology of Mycobacterium bovis in wild deer and feral pigs and their roles in the establishment and spread of bovine tuberculosis in New Zealand wildlife.

    Science.gov (United States)

    Nugent, G; Gortazar, C; Knowles, G

    2015-06-01

    In New Zealand, wild deer and feral pigs are assumed to be spillover hosts for Mycobacterium bovis, and so are not targeted in efforts aimed at locally eradicating bovine tuberculosis (TB) from possums (Trichosurus vulpecula), the main wildlife host. Here we review the epidemiology of TB in deer and pigs, and assess whether New Zealand's TB management programme could be undermined if these species sometimes achieve maintenance host status. In New Zealand, TB prevalences of up to 47% have been recorded in wild deer sympatric with tuberculous possums. Patterns of lesion distribution, age-specific prevalences and behavioural observations suggest that deer become infected mainly through exposure to dead or moribund possums. TB can progress rapidly in some deer (sentinels for detecting TB in wildlife. It is unlikely that wild deer and feral pigs act as maintenance hosts anywhere in New Zealand, because unrestricted year-round hunting keeps densities low, with far less aggregation than on New Zealand farms. We conclude that active management of wild deer or feral pigs is not required for local TB eradication in New Zealand. PMID:25295713

  17. 兔脊柱结核模型的构建%Establishment of a rabbit model of spinal tuberculosis with mycobacterium tuberculosis H37Rv

    Institute of Scientific and Technical Information of China (English)

    刘正文; 呼西旦; 付明花; 折胜利; 罗兰; 宋兴华

    2012-01-01

    目的 在兔腰椎椎体钻孔种植人型H37 Rv结核标准菌株构建兔脊柱结核模型.方法 对16只兔腰5椎体上方钻孔,填充明胶海绵,吸附结核菌混悬液0.1 ml(菌量:5g/L)骨蜡封闭钻孔,在术后不同时间段行大体观察、影像学、组织病理学、细菌学等检查.结果 16只大白兔中,其中13只兔腰椎椎体感染结核菌后,局部反应较明显,全身反应较轻,完成实验;3只未感染结核杆菌.在感染的大白兔中,2只出现消瘦、厌食等症状;术后1个月X线阳性4只,术后2个月X线阳性8只、CT阳性12只、MRI阳性13只;感染大白兔的椎体、椎旁软组织切片苏木素-伊红(HE)染色示有脓细胞、类上皮样细胞或坏死灶等形成,对形成的脓肿进行脓液培养显示69.2%结核分枝杆菌生长.建立模型成功率为69.2%.结论 通过在兔脊柱椎体局部进行适当剂量H37 Rv结核标准菌株的种植感染,可构建出与人类脊柱结核病理变化相似的兔脊柱结核.%Objective Drill a hole in the lumbar vertebral of the rabbit and implant human type H37 Rv tuberculosis standard strain in the hole to construct the model of spinal tuberculosis of the rabbit.Methods Drill holes at the top of the 5th anodic lumbar vertebrae in 16 rabbits,and fill the holes with gelatin sponge to adsorb the M.tuberculosis suspension 0.1 ml ( the amount of bacteria:5 g/L).Finally,close the holes with bone wax.After operation,adopt examinations of general observation,Imaging,Histopathology,Bacteriology etc at different time periods.Results 13 rabbits whose lumbar vertebral bodies were infected had obvious partial response and lesser response of whole bodies.3 of the 16 rabbits were not infected by mycobacterium tuberculosis.2 of the infected ones appeared to have weight loss,anorexia and other symptoms.One month after operation,4 rabbits' X-ray findings were positive; 4 months after operation,8 rabbits' X-ray findings were positive,12 rabbits' CT

  18. Structural analysis of the dodecameric proteasome activator PafE in Mycobacterium tuberculosis.

    Science.gov (United States)

    Bai, Lin; Hu, Kuan; Wang, Tong; Jastrab, Jordan B; Darwin, K Heran; Li, Huilin

    2016-04-01

    The human pathogen Mycobacterium tuberculosis (Mtb) requires a proteasome system to cause lethal infections in mice. We recently found that proteasome accessory factor E (PafE, Rv3780) activates proteolysis by the Mtb proteasome independently of adenosine triphosphate (ATP). Moreover, PafE contributes to the heat-shock response and virulence of Mtb Here, we show that PafE subunits formed four-helix bundles similar to those of the eukaryotic ATP-independent proteasome activator subunits of PA26 and PA28. However, unlike any other known proteasome activator, PafE formed dodecamers with 12-fold symmetry, which required a glycine-XXX-glycine-XXX-glycine motif that is not found in previously described activators. Intriguingly, the truncation of the PafE carboxyl-terminus resulted in the robust binding of PafE rings to native proteasome core particles and substantially increased proteasomal activity, suggesting that the extended carboxyl-terminus of this cofactor confers suboptimal binding to the proteasome core particle. Collectively, our data show that proteasomal activation is not limited to hexameric ATPases in bacteria. PMID:27001842

  19. Mutational analysis of the respiratory nitrate transporter NarK2 of Mycobacterium tuberculosis.

    Directory of Open Access Journals (Sweden)

    Michelle M Giffin

    Full Text Available Mycobacterium tuberculosis induces nitrate reductase activity in response to decreasing oxygen levels. This is due to regulation of both the transcription and the activity of the nitrate transporter NarK2. A model of NarK2 structure is proposed containing 12 membrane spanning regions consistent with other members of the major facilitator superfamily. The role of the proton gradient was determined by exposing M. tuberculosis to uncouplers. Nitrite production decreased indicating that the importation of nitrate involved an H(+/nitrate symporter. The addition of nitrite before nitrate had no effect, suggesting no role for a nitrate/nitrite antiporter. In addition the NarK2 knockout mutant showed no defect in nitrite export. NarK2 is proposed to be a Type I H(+/nitrate symporter. Site directed mutagenesis was performed changing 23 amino acids of NarK2. This allowed the identification of important regions and amino acids of this transporter. Five of these mutants were inactive for nitrate transport, seven produced reduced activity and eleven mutants retained wild type activity. NarK2 is inactivated in the presence of oxygen by an unknown mechanism. However none of the mutants, including those with mutated cysteines, were altered in their response to oxygen levels. The assimilatory nitrate transporter NasA of Bacillus subtilis was expressed in the M. tuberculosis NarK2 mutant. It remained active during aerobic incubation showing that the point of oxygen control is NarK2.

  20. Overexpression, purification and crystallographic analysis of a unique adenosine kinase from Mycobacterium tuberculosis

    International Nuclear Information System (INIS)

    Adenosine kinase from M. tuberculosis has been overexpressed, purified and crystallized in the presence of adenosine. Structure determination using molecular replacement with diffraction data collected at 2.2 Å reveals a dimeric structure. Adenosine kinase from Mycobacterium tuberculosis is the only prokaryotic adenosine kinase that has been isolated and characterized. The enzyme catalyzes the phosphorylation of adenosine to adenosine monophosphate and is involved in the activation of 2-methyladenosine, a compound that has demonstrated selective activity against M. tuberculosis. The mechanism of action of 2-methyladenosine is likely to be different from those of current tuberculosis treatments and this compound (or other adenosine analogs) may prove to be a novel therapeutic intervention for this disease. The M. tuberculosis adenosine kinase was overexpressed in Escherichia coli and the enzyme was purified with activity comparable to that reported previously. The protein was crystallized in the presence of adenosine using the vapour-diffusion method. The crystals diffracted X-rays to high resolution and a complete data set was collected to 2.2 Å using synchrotron radiation. The crystal belonged to space group P3121, with unit-cell parameters a = 70.2, c = 111.6 Å, and contained a single protein molecule in the asymmetric unit. An initial structural model of the protein was obtained by the molecular-replacement method, which revealed a dimeric structure. The monomers of the dimer were related by twofold crystallographic symmetry. An understanding of how the M. tuberculosis adenosine kinase differs from the human homolog should aid in the design of more potent and selective antimycobacterial agents that are selectively activated by this enzyme

  1. Quantitative proteomic analysis of ofloxacin resistant and sensitive clinical isolates of Mycobacterium tuberculosis

    Directory of Open Access Journals (Sweden)

    Xiang-yu HUANG

    2014-10-01

    Full Text Available Objective To identify the proteins related to ofloxacin (OFX resistance of Mycobacterium tuberculosis (MTB. Methods Standard MTB H37Rv strain, clinical isolates of OFX resistant strain (OFXR and sensitive strain (OFXS were obtained from the Chinese Center for Disease Control and Prevention, and they were cultured in Sauton's medium, and then inactivated by 60Co. Whole cellular proteins were extracted from OFXR, OFXS and H37Rv strain of MTB, respectively. The peptides were labeled, separated and identified by isobaric tags of relative and absolute quantitation (iTRAQ combined with Nano LCMS/MS technology. Results One hundred and seventy-five and 134 differential expression proteins were identified in MTB OFXR compared with MTB OFXS and H37Rv, respectively. One hundred and four common differential expression proteins were identified in MTB OFXR compared with both MTB OFXS and H37Rv. The isoelectric point and theoretic relative molecular mass of differential expression proteins were widely distributed. The majority of the common differential expression proteins were involved in intermediary metabolism, respiration, and lipid metabolism. Twelve common differential expression proteins showed significant differences (the ratio>1.2 or <0.55 in MTB OFXR, including Rv0106, Rv0895, Rv2185c, Rv3248c and Rv3841 up-regulation and Rv2524c, Rv2986c, Rv3118 and Rv3597c down-regulation. Conclusion iTRAQ has been used to identify the common differential expression proteins in MTB OFXR compared with both MTB OFXS and H37Rv, which provides a basis for further study of the mechanism of OFX-resistance. DOI: 10.11855/j.issn.0577-7402.2014.09.06

  2. Serial image analysis of Mycobacterium tuberculosis colony growth reveals a persistent subpopulation in sputum during treatment of pulmonary TB

    Science.gov (United States)

    Barr, David A.; Kamdolozi, Mercy; Nishihara, Yo; Ndhlovu, Victor; Khonga, Margaret; Davies, Geraint R.; Sloan, Derek J.

    2016-01-01

    Summary Faster elimination of drug tolerant ‘persister’ bacteria may shorten treatment of tuberculosis (TB) but no method exists to quantify persisters in clinical samples. We used automated image analysis to assess whether studying growth characteristics of individual Mycobacterium tuberculosis colonies from sputum on solid media during early TB treatment facilitates ‘persister’ phenotyping. As Time to Detection (TTD) in liquid culture inversely correlates with total bacterial load we also evaluated the relationship between individual colony growth parameters and TTD. Sputum from TB patients in Malawi was prepared for solid and liquid culture after 0, 2 and 4 weeks of treatment. Serial photography of agar plates was used to measure time to appearance (lag time) and radial growth rate for each colony. Mixed-effects modelling was used to analyse changing growth characteristics from serial samples. 20 patients had colony measurements recorded at ≥1 time-point. Overall lag time increased by 6.5 days between baseline and two weeks (p = 0.0001). Total colony count/ml showed typical biphasic elimination, but long lag time colonies (>20days) had slower, monophasic decline. TTD was associated with minimum lag time (time to appearance of first colony1). Slower elimination of long lag time colonies suggests that these may represent a persister subpopulation of bacilli. PMID:27156626

  3. Overexpression, purification, crystallization and preliminary X-ray analysis of Rv2780 from Mycobacterium tuberculosis H37Rv

    Energy Technology Data Exchange (ETDEWEB)

    Tripathi, Sarvind Mani; Ramachandran, Ravishankar, E-mail: r-ravishankar@cdri.res.in [Molecular and Structural Biology Division, Central Drug Research Institute, PO Box 173, Chattar Manzil, Mahatma Gandhi Marg, Lucknow 226001 (India)

    2008-05-01

    Rv2780, an alanine dehydrogenase from M. tuberculosis, has been crystallized in apo and NAD/pyruvate-bound forms. Preliminary crystallographic analysis shows that there is a hexamer and trimer in the asymmetric units of the apo and ternary complex forms, respectively. Rv2780, an alanine dehydrogenase from Mycobacterium tuberculosis (MtAlaDH), catalyzes the NAD-dependent interconversion of alanine and pyruvate. Alanine dehydrogenase is released into the culture medium in substantial amounts by virulent strains of mycobacteria and is not found in the vaccine strain of tuberculosis. Crystals of recombinant MtAlaDH were grown from 2 M ammonium sulfate solution at ∼12 mg ml{sup −1} protein concentration in two crystal forms which occur in the presence and absence of NAD/pyruvate, respectively. Diffraction data extending to 2.6 Å were collected at room temperature from both apo and ternary complex crystals. Crystals of the apoenzyme have unit-cell parameters a = 173.89, b = 127.07, c = 135.95 Å. They are rod-like in shape and belong to space group C2. They contain a hexamer in the asymmetric unit. Crystals of the ternary complex belong to space group P4{sub 3}2{sub 1}2 and have unit-cell parameters a = b = 88.99, c = 373.85 Å. There are three subunits in the asymmetric unit of the holoenzyme crystals.

  4. Molecular typing of Argentinian Mycobacterium avium subsp. paratuberculosis isolates by multiple-locus variable number-tandem repeat analysis

    Directory of Open Access Journals (Sweden)

    Andrea Gioffré

    2015-06-01

    Full Text Available Multiple-locus variable number-tandem repeat analysis (MLVA of Mycobacterium avium subspecies paratuberculosis (MAP isolates may contribute to the knowledge of strain diversity in Argentina. Although the diversity of MAP has been previously investigated in Argentina using IS900-RFLP, a small number of isolates were employed, and a low discriminative power was reached. The aim of the present study was to test the genetic diversity among MAP isolates using an MLVA approach based on 8 repetitive loci. We studied 97 isolates from cattle, goat and sheep and could describe 7 different patterns: INMV1, INMV2, INMV11, INMV13, INMV16, INMV33 and one incomplete pattern. INMV1 and INMV2 were the most frequent patterns, grouping 76.3% of the isolates. We were also able to demonstrate the coexistence of genotypes in herds and co-infection at the organism level. This study shows that all the patterns described are common to those described in Europe, suggesting an epidemiological link between the continents.

  5. Molecular typing of Argentinian Mycobacterium avium subsp. paratuberculosis isolates by multiple-locus variable number-tandem repeat analysis.

    Science.gov (United States)

    Gioffré, Andrea; Correa Muñoz, Magnolia; Alvarado Pinedo, María F; Vaca, Roberto; Morsella, Claudia; Fiorentino, María Andrea; Paolicchi, Fernando; Ruybal, Paula; Zumárraga, Martín; Travería, Gabriel E; Romano, María Isabel

    2015-06-01

    Multiple-locus variable number-tandem repeat analysis (MLVA) of Mycobacterium avium subspecies paratuberculosis (MAP) isolates may contribute to the knowledge of strain diversity in Argentina. Although the diversity of MAP has been previously investigated in Argentina using IS900-RFLP, a small number of isolates were employed, and a low discriminative power was reached. The aim of the present study was to test the genetic diversity among MAP isolates using an MLVA approach based on 8 repetitive loci. We studied 97 isolates from cattle, goat and sheep and could describe 7 different patterns: INMV1, INMV2, INMV11, INMV13, INMV16, INMV33 and one incomplete pattern. INMV1 and INMV2 were the most frequent patterns, grouping 76.3% of the isolates. We were also able to demonstrate the coexistence of genotypes in herds and co-infection at the organism level. This study shows that all the patterns described are common to those described in Europe, suggesting an epidemiological link between the continents. PMID:26273274

  6. Mutational analysis of the (p)ppGpp synthetase activity of the Rel enzyme of Mycobacterium tuberculosis.

    Science.gov (United States)

    Bag, Satyabrata; Das, Bhabatosh; Dasgupta, Shreya; Bhadra, Rupak K

    2014-08-01

    Rel(Mtb), a GTP pyrophosphokinase encoded by the Mycobacterium tuberculosis (Mtb) genome, catalyzes synthesis of (p)ppGpp from ATP and GDP(GTP) and its hydrolysis to GDP(GTP) and pyrophosphate to mediate stringent response, which helps bacteria to survive during nutrient limitation. Like other members of Rel_Spo homologs, Rel(Mtb) has four distinct domains: HD, Rel_Spo (RSD), TGS and ACT. The N-terminal HD and RSD are responsible for (p)ppGpp hydrolysis and synthesis, respectively. In this study, we have dissected the rel(Mtb) gene function and determined the minimal region essential for (p)ppGpp synthetic activity. The Rel(Mtb) and its truncated derivatives were expressed from an arabinose inducible promoter (P(BAD)), and in vivo functional analyses were done in a (p)ppGpp null Escherichia coli strain. Our results indicate that only 243 amino acids (188-430 residues) containing fragment are sufficient for Rel(Mtb) (p)ppGpp synthetic activity. The results were further confirmed by in vitro assays using purified proteins. We further characterized the RSD of Rel(Mtb) by substituting several conserved amino acids with structurally related residues and identified six such residues, which appeared to be critical for maintaining its catalytic activity. Furthermore, we have also extended our analysis to an RSD encoding gene rv1366 of Mtb, and experimental results indicated that the encoded protein Rv1366 is unable to synthesize (p)ppGpp.

  7. 270例结核分支杆菌药敏试验结果分析%Analysis of 270 cases of mycobacterium tuberculosis drug susceptility test results

    Institute of Scientific and Technical Information of China (English)

    贾琳; 张红; 魏成翠

    2013-01-01

    Objective Through the 270 cases of mycobacterium tuberculosis drug susceptility test results of the analysis ,to explore the incidence of anti-tuberculosis drug resistance and characteristic ,provide the basis for drug-resistant tuberculosis treatment and prevention. Methords 270 cases of mycobacterium tuberculosis by absolute concentration method for 12kinds of drug sensitive test. Results 270 cases of mycobacterium tuberculosis were detected 142 cases of drug-resistant(52.6%) ,among them multidrug resistance(MDR-TB) 83 cases(30.7%). Conclusions mycobacterium tuberculosis drug-resistant situation is more serious ,should further strengthen clinical treatment and standard medication ,control tuberculosis drug-resistant increase.%  目的通过对270例结核分支杆菌药敏试验结果的分析,探讨抗结核药物耐药性的发生率和特点,为耐药结核病预防和治疗提供依据。方法采用绝对浓度法对270例结核分支杆菌进行12种药物的敏感试验。结果270例结核分支杆菌共检出耐药菌142例(52.6%),其中耐多药结核(MDR-TB)83例(30.7%)。结论结核分支杆菌耐药状况比较严重,应进一步加强结核病的临床治疗和规范用药,控制耐药菌增加。

  8. Economic analysis of Mycobacterium avium subspecies paratuberculosis vaccines in dairy herds.

    Science.gov (United States)

    Cho, J; Tauer, L W; Schukken, Y H; Gómez, M I; Smith, R L; Lu, Z; Grohn, Y T

    2012-04-01

    Johne's disease, or paratuberculosis, is a chronic infectious enteric disease of ruminants, caused by infection with Mycobacterium avium ssp. paratuberculosis (MAP). Given the absence of a fail-safe method of prevention or a cure, Johne's disease can inflict significant economic loss on the US dairy industry, with an estimated annual cost of over $200 million. Currently available MAP control strategies include management measures to improve hygiene, culling MAP serologic- or fecal-positive adult cows, and vaccination. Although the 2 first control strategies have been reported to be effective in reducing the incidence of MAP infection, the changes in herd management needed to conduct these control strategies require significant effort on the part of the dairy producer. On the other hand, vaccination is relatively simple to apply and requires minor changes in herd management. Despite these advantages, only 5% of US dairy operations use vaccination to control MAP. This low level of adoption of this technology is due to limited information on its cost-effectiveness and efficacy and some important inherent drawbacks associated with current MAP vaccines. This study investigates the epidemiological effect and economic values of MAP vaccines in various stages of development. We create scenarios for the potential epidemiological effects of MAP vaccines, and then estimate economically justifiable monetary values at which vaccines become economically beneficial to dairy producers such that a net present value (NPV) of a farm's net cash flow can be higher than the NPV of a farm using no control or alternative nonvaccine controls. Any vaccination with either low or high efficacy considered in this study yielded a higher NPV compared with a no MAP control. Moreover, high-efficacy vaccines generated an even higher NPV compared with alternative controls, making vaccination economically attractive. Two high-efficacy vaccines were particularly effective in MAP control and NPV

  9. Integrated gene co-expression network analysis in the growth phase of Mycobacterium tuberculosis reveals new potential drug targets.

    Science.gov (United States)

    Puniya, Bhanwar Lal; Kulshreshtha, Deepika; Verma, Srikant Prasad; Kumar, Sanjiv; Ramachandran, Srinivasan

    2013-11-01

    We have carried out weighted gene co-expression network analysis of Mycobacterium tuberculosis to gain insights into gene expression architecture during log phase growth. The differentially expressed genes between at least one pair of 11 different M. tuberculosis strains as source of biological variability were used for co-expression network analysis. This data included genes with highest coefficient of variation in expression. Five distinct modules were identified using topological overlap based clustering. All the modules together showed significant enrichment in biological processes: fatty acid biosynthesis, cell membrane, intracellular membrane bound organelle, DNA replication, Quinone biosynthesis, cell shape and peptidoglycan biosynthesis, ribosome and structural constituents of ribosome and transposition. We then extracted the co-expressed connections which were supported either by transcriptional regulatory network or STRING database or high edge weight of topological overlap. The genes trpC, nadC, pitA, Rv3404c, atpA, pknA, Rv0996, purB, Rv2106 and Rv0796 emerged as top hub genes. After overlaying this network on the iNJ661 metabolic network, the reactions catalyzed by 15 highly connected metabolic genes were knocked down in silico and evaluated by Flux Balance Analysis. The results showed that in 12 out of 15 cases, in 11 more than 50% of reactions catalyzed by genes connected through co-expressed connections also had altered fluxes. The modules 'Turquoise', 'Blue' and 'Red' also showed enrichment in essential genes. We could map 152 of the previously known or proposed drug targets in these modules and identified 15 new potential drug targets based on their high degree of co-expressed connections and strong correlation with module eigengenes.

  10. Establishment and validation of a guinea pig model of latent Mycobacterium tuberculosis infection%结核分枝杆菌潜伏感染豚鼠模型的建立和验证

    Institute of Scientific and Technical Information of China (English)

    卢锦标; 邓海清; 陈保文; 都伟欣; 杨蕾; 沈小兵; 苏城; 徐苗; 王国治

    2013-01-01

    目的:建立结核分枝杆菌(M ycobacterium tuberclu osis,Mtb)潜伏感染治疗用疫苗的豚鼠评价模型。方法豚鼠皮下攻毒5.0×103 CFU Mtb,于不同时间点用0.5μg重组结核杆菌ESAT6-CFP10蛋白对豚鼠皮试,观察皮试阳转情况。攻毒2周后,模型组豚鼠采用5 mg异烟肼灌胃治疗,每周3次,持续4周;对照组豚鼠不做化疗。在Mtb感染后的第6周,分别取未经异烟肼治疗的豚鼠和经异烟肼治疗的豚鼠解剖,比较豚鼠的肝、脾和肺的脏器综合病变指数以及脾活菌数( log10 CFU)。根据上述结果建立豚鼠的潜伏感染模型,并用两个参考疫苗对该模型进行验证。结果豚鼠攻毒Mtb 2周后,EC皮试反应全部转阳,皮试反应大小为(19.9±3.0) mm。模型组豚鼠经4周异烟肼治疗后,脏器综合病变指数为0,脾未分离出活菌数。对照组豚鼠不经化疗,脏器综合病变指数为38.8±16.5,脾活菌数为(5.1±0.3)log10 CFU。停药后,模型组豚鼠结核病自然复发,在两次参考疫苗的验证中,脏器综合病变指数分别为48.5±23.9和51.30±23.41,脾活菌数分别为(4.5±1.3)和(4.2±1.1) log10 CFU,肺部活菌数分别为(4.1±1.2)和(3.4±1.3) log10 CFU。结论 Mtb攻毒豚鼠后,异烟肼化疗可抑制Mtb增殖,成功建立豚鼠的Mtb潜伏感染模型。该模型重复性好,可用于潜伏感染治疗用疫苗的评价。%Objective To establish a guinea pig model of latent Mycobacterium tuberculosis infec-tion for evaluating the effects of therapeutic vaccines .Methods Guinea pigs were subcutaneously inocula-ted with 5.0×103 CFU Mtb.The skin test was performed with 0.5μg recombinant ESAT6-CFP10 protein to detect positive conversion rates at different time points .Two weeks after Mtb inoculation , guinea pigs in model group received 5 mg isoniazid treatment ( three times a week for four weeks

  11. Central metabolism in Mycobacterium smegmatis during the transition from O2-rich to O2-poor conditions as studied by isotopomer-assisted metabolite analysis.

    Science.gov (United States)

    Tang, Yinjie J; Shui, Wenqing; Myers, Samuel; Feng, Xueyang; Bertozzi, Carolyn; Keasling, Jay D

    2009-08-01

    Isotopomer-assisted metabolite analysis was used to investigate the central metabolism of Mycobacterium smegmatis and its transition from normal growth to a non-replicating state under a hypoxic environment. Tween 80 significantly promoted aerobic growth by improving O(2) transfer, while only small amount was degraded and metabolized via the TCA cycle for biomass synthesis. As the bacillus encountered hypoxic stress, isotopomer analysis suggested: (1) isocitrate lyase activity increased, which further induced glyoxylate pathway and glycine dehydrogenase for replenishing NAD(+); (2) the relative amount of acetyl-CoA entering the TCA cycle was doubled, whereas little entered the glycolytic and pentose phosphate pathways. PMID:19357814

  12. Analysis on Establishing Urban Cemetery Planning and Compiling System

    Institute of Scientific and Technical Information of China (English)

    Kun; YANG; Xiaogang; CHEN

    2015-01-01

    Currently,there are many problems in construction of urban cemetery like improper location,low land utilization,backward greening facilities and imperfect cemetery management,which have greatly affected people’s normal production and life. This article discusses the establishment of a sustainable city cemetery planning and compiling system from three levels of " macro-view,medium-view and micro-view" in order to perfect the present cemetery system.

  13. Análisis transcripcional de la región genética RvD1 de Mycobacterium bovis Transcriptional analysis of genetic region RvD1 of Mycobacterium bovis

    Directory of Open Access Journals (Sweden)

    del Portillo Patricia

    2004-12-01

    Full Text Available Mycobacterium bovis comparte una identidad del 99,9% con los genomas de M. tuberculosis, M. africanum y M. microti. Dentro del 0,1% de esta diferencia se encuentran dos regiones genéticas propias de M. bovis: RvD1 y RvD2, las cuales se encuentran delecionadas del genoma de M. tuberculosis H37Rv y, según el análisis bioin-formático, contienen probables marcos abiertos de lectura (Open Reading Frames: ORF. Con el fin de deter­minar si la región RvD1, transcribe los ORF predichos por bioinformática: ORF1, ORF2 y Rv2024, se extrajeron muestras de ARN total de M. bovis BCG Pasteur, en diferentes puntos de una curva de crecimiento micobacteriano, las cuales fueron analizadas mediante la técnica de Transcripción Reversa y Reacción en Ca­dena de la Polimerasa (RTq-PCR en tiempo real. Los hallazgos obtenidos en esta cinética de transcripción por RTq-PCR en tiempo real demostraron que los probables marcos de lectura abiertos ORF1, ORF2 y Rv2024 de la región RvD1 de M. bovis, sí se transcriben y lo hacen de manera constitutiva, hecho que no había sido repor­tado. Los resultados de esta investigación sirven como un primer paso para determinar la función que desem­peña la región RvD1 de M. bovis, y su posible papel en la patogénesis y en la interacción huésped-patógeno de la tuberculosis bovina y humana. Palabras clave: Mycobacterium bovis, BCG, RNA, RT-PCR, RvD1.Mycobacterium bovis, shares 99.9% of genomic identity with M. tuberculosis, M. africanum and M. microti. Within this 0.1 % of difference, there are two genetic regions characteristics of M. bovis that are deleted in M. tuberculo­sis H37Rv: RvD1 and RvD2. According to bioinformatic analysis, these regions contain Open Reading Frames (ORFs. With the purpose of determining if the RvD1 region transcribes the ORFs predicted by bioinformatics (ORF1, ORF2 and Rv2024; total RNA was extracted from a culture of M. bovis BCG Pasteur, at different time points along the growth curve

  14. Whole-Genome Sequencing Analysis of Serially Isolated Multi-Drug and Extensively Drug Resistant Mycobacterium tuberculosis from Thai Patients.

    Science.gov (United States)

    Faksri, Kiatichai; Tan, Jun Hao; Disratthakit, Areeya; Xia, Eryu; Prammananan, Therdsak; Suriyaphol, Prapat; Khor, Chiea Chuen; Teo, Yik-Ying; Ong, Rick Twee-Hee; Chaiprasert, Angkana

    2016-01-01

    Multi-drug and extensively drug-resistant tuberculosis (MDR and XDR-TB) are problems that threaten public health worldwide. Only some genetic markers associated with drug-resistant TB are known. Whole-genome sequencing (WGS) is a promising tool for distinguishing between re-infection and persistent infection in isolates taken at different times from a single patient, but has not yet been applied in MDR and XDR-TB. We aim to detect genetic markers associated with drug resistance and distinguish between reinfection and persistent infection from MDR and XDR-TB patients based on WGS analysis. Samples of Mycobacterium tuberculosis (n = 7), serially isolated from 2 MDR cases and 1 XDR-TB case, were retrieved from Siriraj Hospital, Bangkok. The WGS analysis used an Illumina Miseq sequencer. In cases of persistent infection, MDR-TB isolates differed at an average of 2 SNPs across the span of 2-9 months whereas in the case of reinfection, isolates differed at 61 SNPs across 2 years. Known genetic markers associated with resistance were detected from strains susceptible to streptomycin (2/7 isolates), p-aminosalicylic acid (3/7 isolates) and fluoroquinolone drugs. Among fluoroquinolone drugs, ofloxacin had the highest phenotype-genotype concordance (6/7 isolates), whereas gatifloxcain had the lowest (3/7 isolates). A putative candidate SNP in Rv2477c associated with kanamycin and amikacin resistance was suggested for further validation. WGS provided comprehensive results regarding molecular epidemiology, distinguishing between persistent infection and reinfection in M/XDR-TB and potentially can be used for detection of novel mutations associated with drug resistance. PMID:27518818

  15. Molecular characterization of Mycobacterium tuberculosis isolates from Tehran, Iran by restriction fragment length polymorphism analysis and spoligotyping.

    Science.gov (United States)

    Feyisa, Seifu Gizaw; Haeili, Mehri; Zahednamazi, Fatemeh; Mosavari, Nader; Taheri, Mohammad Mohammad; Hamzehloo, Gholamreza; Zamani, Samin; Feizabadi, Mohammad Mehdi

    2016-04-01

    INTRODUCTION Characterization of Mycobacterium tuberculosis (MTB) isolates by DNA fingerprinting has contributed to tuberculosis (TB) control. The aim of this study was to determine the genetic diversity of MTB isolates from Tehran province in Iran. METHODS MTB isolates from 60 Iranian and 10 Afghan TB patients were fingerprinted by standard IS6110-restriction fragment length polymorphism (RFLP) analysis and spoligotyping. RESULTS The copy number of IS6110 ranged from 10-24 per isolate. The isolates were classified into 22 clusters showing ≥ 80% similarity by RFLP analysis. Fourteen multidrug-resistant (MDR) isolates were grouped into 4 IS6110-RFLP clusters, with 10 isolates [71% (95% CI: 45-89%)] in 1 cluster, suggesting a possible epidemiological linkage. Eighteen Iranian isolates showed ≥ 80% similarity with Afghan isolates. There were no strains with identical fingerprints. Spoligotyping of 70 isolates produced 23 distinct patterns. Sixty (85.7%) isolates were grouped into 13 clusters, while the remaining 10 isolates (14.2%) were not clustered. Ural (formerly Haarlem4) (n = 22, 31.4%) was the most common family followed by Central Asian strain (CAS) (n = 18, 25.7%) and T (n = 9, 12.8%) families. Only 1strain was characterized as having the Beijing genotype. Among 60 Iranian and 10 Afghan MTB isolates, 25% (95% CI: 16-37) and 70% (95% CI: 39-89) were categorized as Ural lineage, respectively. CONCLUSIONS A higher prevalence of Ural family MTB isolates among Afghan patients than among Iranian patients suggests the possible transmission of this lineage following the immigration of Afghans to Iran.

  16. Rapid detection of isoniazid resistance in Mycobacterium tuberculosis isolates by use of real-time-PCR-based melting curve analysis.

    Science.gov (United States)

    Hu, Siyu; Li, Guoli; Li, Hui; Liu, Xiaoli; Niu, Jianjun; Quan, Shengmao; Wang, Feng; Wen, Huixin; Xu, Ye; Li, Qingge

    2014-05-01

    The MeltPro TB/INH assay, recently approved by the Chinese Food and Drug Administration, is a closed-tube, dual-color, melting curve analysis-based, real-time PCR test specially designed to detect 30 isoniazid (INH) resistance mutations in katG position 315 (katG 315), the inhA promoter (positions -17 to -8), inhA position 94, and the ahpC promoter (positions -44 to -30 and -15 to 3) of Mycobacterium tuberculosis. Here we evaluated both the analytical performance and clinical performance of this assay. Analytical studies with corresponding panels demonstrated that the accuracy for detection of different mutation types (10 wild-type samples and 12 mutant type samples), the limit of detection (2×10(3) to 2×10(4) bacilli/ml), reproducibility (standard deviation [SD], real-time PCR machines, with the shortest running time (105 min) obtained with the LightCycler 480 II. Clinical studies enrolled 1,096 clinical isolates collected from three geographically different tuberculosis centers, including 437 INH-resistant isolates and 659 INH-susceptible isolates characterized by traditional drug susceptibility testing on Löwenstein-Jensen solid medium. The clinical sensitivity and specificity of the MeltPro TB/INH assay were 90.8% and 96.4%, respectively. DNA sequencing analysis showed that, except for the 5 mutants outside the detection range of the MeltPro assay, a concordance rate between the two methods of 99.1% (457/461) was obtained. Among the 26 mutation types detected, katG S315T (AGC→ACC), inhA -15C→T, katG S315N (AGC→AAC), and ahpC promoter -10C→T accounted for more than 90%. Overall, the MeltPro TB/INH assay represents a reliable and rapid tool for the detection of INH resistance in clinical isolates.

  17. High accuracy mass spectrometry analysis as a tool to verify and improve gene annotation using Mycobacterium tuberculosis as an example

    Directory of Open Access Journals (Sweden)

    Prasad Swati

    2008-07-01

    Full Text Available Abstract Background While the genomic annotations of diverse lineages of the Mycobacterium tuberculosis complex are available, divergences between gene prediction methods are still a challenge for unbiased protein dataset generation. M. tuberculosis gene annotation is an example, where the most used datasets from two independent institutions (Sanger Institute and Institute of Genomic Research-TIGR differ up to 12% in the number of annotated open reading frames, and 46% of the genes contained in both annotations have different start codons. Such differences emphasize the importance of the identification of the sequence of protein products to validate each gene annotation including its sequence coding area. Results With this objective, we submitted a culture filtrate sample from M. tuberculosis to a high-accuracy LTQ-Orbitrap mass spectrometer analysis and applied refined N-terminal prediction to perform comparison of two gene annotations. From a total of 449 proteins identified from the MS data, we validated 35 tryptic peptides that were specific to one of the two datasets, representing 24 different proteins. From those, 5 proteins were only annotated in the Sanger database. In the remaining proteins, the observed differences were due to differences in annotation of transcriptional start sites. Conclusion Our results indicate that, even in a less complex sample likely to represent only 10% of the bacterial proteome, we were still able to detect major differences between different gene annotation approaches. This gives hope that high-throughput proteomics techniques can be used to improve and validate gene annotations, and in particular for verification of high-throughput, automatic gene annotations.

  18. Molecular analysis and MIRU-VNTR typing of Mycobacterium avium subsp. avium, 'hominissuis' and silvaticum strains of veterinary origin.

    Science.gov (United States)

    Rónai, Zsuzsanna; Csivincsik, Ágnes; Dán, Ádám; Gyuranecz, Miklós

    2016-06-01

    Besides Mycobacterium avium subsp. paratuberculosis (MAP), M. avium subsp. avium (MAA), M. avium subsp. silvaticum (MAS), and 'M. avium subsp. hominissuis' (MAH) are equally important members of M. avium complex, with worldwide distribution and zoonotic potential. Genotypic discrimination is a prerequisite to epidemiological studies which can facilitate disease prevention through revealing infection sources and transmission routes. The primary aim of this study was to identify the genetic diversity within 135 MAA, 62 MAS, and 84 MAH strains isolated from wild and domestic mammals, reptiles and birds. Strains were tested for the presence of large sequence polymorphism LSP(A)17 and were submitted to Mycobacterial interspersed repetitive units-variable-number tandem repeat (MIRU-VNTR) analysis at 8 loci, including MIRU1, 2, 3, and 4, VNTR25, 32, and 259, and MATR9. In 12 strains hsp65 sequence code type was also determined. LSP(A)17 was present only in 19.9% of the strains. All LSP(A)17 positive strains belonged to subspecies MAH. The discriminatory power of the MIRU-VNTR loci set used reached 0.9228. Altogether 54 different genotypes were detected. Within MAH, MAA, and MAS strains 33, 16, and 5 different genotypes were observed. The described genotypes were not restricted to geographic regions or host species, but proved to be subspecies specific. Our knowledge about MAS is limited due to isolation and identification difficulties. This is the first study including a large number of MAS field strains. Our results demonstrate the high diversity of MAH and MAA strains and the relative uniformity of MAS strains. PMID:26964909

  19. Systems biology analysis of gene expression during in vivo Mycobacterium avium paratuberculosis enteric colonization reveals role for immune tolerance.

    Directory of Open Access Journals (Sweden)

    Sangeeta Khare

    Full Text Available Survival and persistence of Mycobacterium avium subsp. paratuberculosis (MAP in the intestinal mucosa is associated with host immune tolerance. However, the initial events during MAP interaction with its host that lead to pathogen survival, granulomatous inflammation, and clinical disease progression are poorly defined. We hypothesize that immune tolerance is initiated upon initial contact of MAP with the intestinal Peyer's patch. To test our hypothesis, ligated ileal loops in neonatal calves were infected with MAP. Intestinal tissue RNAs were collected (0.5, 1, 2, 4, 8 and 12 hrs post-infection, processed, and hybridized to bovine gene expression microarrays. By comparing the gene transcription responses of calves infected with the MAP, informative complex patterns of expression were clearly visible. To interpret these complex data, changes in the gene expression were further analyzed by dynamic Bayesian analysis, and genes were grouped into the specific pathways and gene ontology categories to create a holistic model. This model revealed three different phases of responses: i early (30 min and 1 hr post-infection, ii intermediate (2, 4 and 8 hrs post-infection, and iii late (12 hrs post-infection. We describe here the data that include expression profiles for perturbed pathways, as well as, mechanistic genes (genes predicted to have regulatory influence that are associated with immune tolerance. In the Early Phase of MAP infection, multiple pathways were initiated in response to MAP invasion via receptor mediated endocytosis and changes in intestinal permeability. During the Intermediate Phase, perturbed pathways involved the inflammatory responses, cytokine-cytokine receptor interaction, and cell-cell signaling. During the Late Phase of infection, gene responses associated with immune tolerance were initiated at the level of T-cell signaling. Our study provides evidence that MAP infection resulted in differentially regulated genes, perturbed

  20. targetTB: A target identification pipeline for Mycobacterium tuberculosis through an interactome, reactome and genome-scale structural analysis

    Directory of Open Access Journals (Sweden)

    Chandra Nagasuma

    2008-12-01

    Full Text Available Abstract Background Tuberculosis still remains one of the largest killer infectious diseases, warranting the identification of newer targets and drugs. Identification and validation of appropriate targets for designing drugs are critical steps in drug discovery, which are at present major bottle-necks. A majority of drugs in current clinical use for many diseases have been designed without the knowledge of the targets, perhaps because standard methodologies to identify such targets in a high-throughput fashion do not really exist. With different kinds of 'omics' data that are now available, computational approaches can be powerful means of obtaining short-lists of possible targets for further experimental validation. Results We report a comprehensive in silico target identification pipeline, targetTB, for Mycobacterium tuberculosis. The pipeline incorporates a network analysis of the protein-protein interactome, a flux balance analysis of the reactome, experimentally derived phenotype essentiality data, sequence analyses and a structural assessment of targetability, using novel algorithms recently developed by us. Using flux balance analysis and network analysis, proteins critical for survival of M. tuberculosis are first identified, followed by comparative genomics with the host, finally incorporating a novel structural analysis of the binding sites to assess the feasibility of a protein as a target. Further analyses include correlation with expression data and non-similarity to gut flora proteins as well as 'anti-targets' in the host, leading to the identification of 451 high-confidence targets. Through phylogenetic profiling against 228 pathogen genomes, shortlisted targets have been further explored to identify broad-spectrum antibiotic targets, while also identifying those specific to tuberculosis. Targets that address mycobacterial persistence and drug resistance mechanisms are also analysed. Conclusion The pipeline developed provides

  1. Transcription analysis of the dnaA gene and oriC region of the chromosome of Mycobacterium smegmatis and Mycobacterium bovis BCG, and its regulation by the DnaA protein.

    Science.gov (United States)

    Salazar, Leiria; Guerrero, Elba; Casart, Yveth; Turcios, Lilia; Bartoli, Fulvia

    2003-03-01

    The regions flanking the Mycobacterium dnaA gene have extensive sequence conservation, and comprise various DnaA boxes. Comparative analysis of the dnaA promoter and oriC region from several mycobacterial species revealed that the localization, spacing and orientation of the DnaA boxes are conserved. Detailed transcriptional analysis in M. smegmatis and M. bovis BCG shows that the dnaN gene of both species and the dnaA gene of M. bovis BCG are transcribed from two promoters, whereas the dnaA gene of M. smegmatis is transcribed from a single promoter. RT-PCR with total RNA showed that dnaA and dnaN were expressed in both species at all growth stages. Analysis of the promoter activity using dnaA-gfp fusion plasmids and DnaA expression plasmids indicates that the dnaA gene is autoregulated, although the degree of transcriptional autorepression was moderate. Transcription was also detected in the vicinity of oriC of M. bovis BCG, but not of M. smegmatis. These results suggest that a more complex transcriptional mechanism may be involved in the slow-growing mycobacteria, which regulates the expression of dnaA and initiation of chromosomal DNA replication.

  2. Expression, purification and preliminary crystallographic analysis of Mycobacterium tuberculosis CysQ, a phosphatase involved in sulfur metabolism

    Energy Technology Data Exchange (ETDEWEB)

    Erickson, Anna I.; Sarsam, Reta D.; Fisher, Andrew J., E-mail: ajfisher@ucdavis.edu

    2014-05-10

    The cysQ gene from Mycobacterium tuberculosis was cloned and the expressed protein, a 3′-phosphoadenosine-5′’-phosphatase, was purified and crystallized. X-ray diffraction data were collected to 1.7 Å resolution.

  3. Comparative analysis of Mycobacterium tuberculosis pe and ppe genes reveals high sequence variation and an apparent absence of selective constraints.

    NARCIS (Netherlands)

    McEvoy, C.R.; Cloete, R.; Müller, B.; Schürch, A.C.; Helden, P.D. van; Gagneux, S.; Warren, R.M.; Gey van Pittius, N.C.

    2012-01-01

    Mycobacterium tuberculosis complex (MTBC) genomes contain 2 large gene families termed pe and ppe. The function of pe/ppe proteins remains enigmatic but studies suggest that they are secreted or cell surface associated and are involved in bacterial virulence. Previous studies have also shown that so

  4. Diagnostic test accuracy of anti-glycopeptidolipid-core IgA antibodies for Mycobacterium avium complex pulmonary disease: systematic review and meta-analysis

    OpenAIRE

    Yuji Shibata; Nobuyuki Horita; Masaki Yamamoto; Toshinori Tsukahara; Hideyuki Nagakura; Ken Tashiro; Hiroki Watanabe; Kenjiro Nagai; Kentaro Nakashima; Ryota Ushio; Misako Ikeda; Atsuya Narita; Akinori Kanai; Takashi Sato; Takeshi Kaneko

    2016-01-01

    Currently, an anti-glycopeptidolipid (GPL)-core IgA antibody assay kit for diagnosing Mycobacterium avium complex (MAC) is commercially available. We conducted this systematic review and meta-analysis to reveal the precise diagnostic accuracy of anti-GPL-core IgA antibodies for MAC pulmonary disease (MAC-PD). We systematically searched reports that could provide data for both sensitivity and specificity by anti-GPL-core IgA antibody for clinically diagnosed MAC-PD. Diagnostic test accuracy wa...

  5. Establishing a framework for comparative analysis of genome sequences

    Energy Technology Data Exchange (ETDEWEB)

    Bansal, A.K.

    1995-06-01

    This paper describes a framework and a high-level language toolkit for comparative analysis of genome sequence alignment The framework integrates the information derived from multiple sequence alignment and phylogenetic tree (hypothetical tree of evolution) to derive new properties about sequences. Multiple sequence alignments are treated as an abstract data type. Abstract operations have been described to manipulate a multiple sequence alignment and to derive mutation related information from a phylogenetic tree by superimposing parsimonious analysis. The framework has been applied on protein alignments to derive constrained columns (in a multiple sequence alignment) that exhibit evolutionary pressure to preserve a common property in a column despite mutation. A Prolog toolkit based on the framework has been implemented and demonstrated on alignments containing 3000 sequences and 3904 columns.

  6. Establishment of a Policy Analysis Capability in Romania

    Directory of Open Access Journals (Sweden)

    Carole M.P. NEVES

    2000-01-01

    Full Text Available The Romanian government, like other governments of former communist countries, emerged from the collapsed Soviet Union ill prepared to confront the complex challenges of governing under a democratic, free market system. At the core of governments that formulate sound public policies, successfully implement programs, and respond effectively to rapidly changing situations is the capacity to carry out independent, high quality research and analysis that results in improved decision making. During the 21st century, the need and acceptance of the incorporation of policy analysis courses in public administration curricula in Romania is expected to grow. In time, educational programs are expected to result in the following consequences: _ Formation of a body of strong independent policy researchers employed by the national and local governments as well as by universities and non-profit institutions _ Utilization of analytical outcomes as tools of political, social and economic improvement by the executive, legislative and judicial branches and by non-profit and private sectors _ Greater public understanding of and participation in public policy processes.

  7. Revisiting the structure of the anti-neoplastic glucans of Mycobacterium bovis Bacille Calmette-Guerin. Structural analysis of the extracellular and boiling water extract-derived glucans of the vaccine substrains.

    Science.gov (United States)

    Dinadayala, Premkumar; Lemassu, Anne; Granovski, Pierre; Cérantola, Stéphane; Winter, Nathalie; Daffé, Mamadou

    2004-03-26

    The attenuated strain of Mycobacterium bovis Bacille Calmette-Guérin (BCG), used worldwide to prevent tuberculosis and leprosy, is also clinically used as an immunotherapeutic agent against superficial bladder cancer. An anti-tumor polysaccharide has been isolated from the boiling water extract of the Tice substrain of BCG and tentatively characterized as consisting primarily of repeating units of 6-linked-glucosyl residues. Mycobacterium tuberculosis and other mycobacterial species produce a glycogen-like alpha-glucan composed of repeating units of 4-linked glucosyl residues substituted at some 6 positions by short oligoglucosyl units that also exhibits an anti-tumor activity. Therefore, the impression prevails that mycobacteria synthesize different types of anti-neoplastic glucans or, alternatively, the BCG substrains are singular in producing a unique type of glucan that may confer to them their immunotherapeutic property. The present study addresses this question through the comparative analysis of alpha-glucans purified from the extracellular materials and boiling water extracts of three vaccine substrains. The polysaccharides were purified, and their structural features were established by mono- and two-dimensional NMR spectroscopy and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry of the enzymatic and chemical degradation products of the purified compounds. The glucans isolated by the two methods from the three substrains of BCG were shown to exhibit identical structural features shared with the glycogen-like alpha-glucan of M. tuberculosis and other mycobacteria. Incidentally, we observed an occasional release of dextrans from Sephadex columns that may explain the reported occurrence of 6-substituted alpha-glucans in mycobacteria.

  8. Diesel engine coolant analysis, new application for established instrumentation

    Energy Technology Data Exchange (ETDEWEB)

    Anderson, D.P.; Lukas, M.; Lynch, B.K. [Spectro Incorporated, Littleton, MA (United States)

    1997-12-31

    Rotating disk electrode (RDE) arc emission spectrometers are user` many commercial, industrial and military laboratories throughout the world to analyze millions of oil and fuel samples each year. In fact, RDE spectrometers have been used exclusively for oil and fuel analysis for so long that it has nearly been forgotten by most practitioners that when RDE spectrometers were first introduced more than 40 years ago, they were routinely used for aqueous samples as well. This presentation reviews early methods of aqueous sample analysis using RDE technology. This presentation also describes recent work to calibrate an RDE spectrometer for both water samples and for engine coolant samples which are a mixture of approximately 50 % water and 50 % ethylene or propylene glycol. Limits of detection determined for aqueous standards are comparable to limits of detection for oil standards. Repeatability of aqueous samples is comparable to the repeatability achieved for oil samples. A comparison of results for coolant samples measured by both inductively coupled plasma (ICP) and rotating disk electrode (RDE) spectrometers is presented. Not surprisingly, RDE results are significantly higher for samples containing particles larger than a few micrometers. Although limits of detection for aqueous samples are not as low as can be achieved using the more modern ICP spectrometric method or the more cumbersome atomic absorption (AA) method, this presentation suggests that RDE spectrometers may be appropriate for certain types of aqueous samples in situations where the more sensitive ICP or AA spectrometers and the laboratory environment and skilled personnel needed for them to operate are not conveniently available. (orig.) 4 refs.

  9. Mycobacterium bovis (Bovine Tuberculosis) in Humans

    Science.gov (United States)

    Mycobacterium bovis (Bovine Tuberculosis) in Humans What is Mycobacterium bovis ? In the United States, the majority of tuberculosis (TB) cases in people are caused by Mycobacterium tuberculosis ( ...

  10. Quantitative Proteomics and Lipidomics Analysis of Endoplasmic Reticulum of Macrophage Infected with Mycobacterium tuberculosis

    Directory of Open Access Journals (Sweden)

    Najmuddin Mohd Saquib

    2015-01-01

    Full Text Available Even though endoplasmic reticulum (ER stress associated with mycobacterial infection has been well studied, the molecular basis of ER as a crucial organelle to determine the fate of Mtb is yet to be established. Here, we have studied the ability of Mtb to manipulate the ultrastructural architecture of macrophage ER and found that the ER-phenotypes associated with virulent (H37Rv and avirulent (H37Ra strains were different: a rough ER (RER with the former against a smooth ER (SER with the later. Further, the functional attributes of these changes were probed by MS-based quantitative proteomics (133 ER proteins and lipidomics (8 phospholipids. Our omics approaches not only revealed the host pathogen cross-talk but also emphasized how precisely Mtb uses proteins and lipids in combination to give rise to characteristic ER-phenotypes. H37Ra-infected macrophages increased the cytosolic Ca2+ levels by attenuating the ATP2A2 protein and simultaneous induction of PC/PE expression to facilitate apoptosis. However, H37Rv inhibited apoptosis and further controlled the expression of EST-1 and AMRP proteins to disturb cholesterol homeostasis resulting in sustained infection. This approach offers the potential to decipher the specific roles of ER in understanding the cell biology of mycobacterial infection with special reference to the impact of host response.

  11. Proteomic analysis of the action of the Mycobacterium ulcerans toxin mycolactone: targeting host cells cytoskeleton and collagen.

    OpenAIRE

    Gama, José B.; Steffen Ohlmeier; Martins, Teresa G.; Fraga, Alexandra G.; Belém Sampaio-Marques; Carvalho, Maria A.; Fernanda Proença; Silva, Manuel T.; Jorge Pedrosa; Paula Ludovico

    2014-01-01

    Buruli ulcer (BU) is a neglected tropical disease caused by Mycobacterium ulcerans. The tissue damage characteristic of BU lesions is known to be driven by the secretion of the potent lipidic exotoxin mycolactone. However, the molecular action of mycolactone on host cell biology mediating cytopathogenesis is not fully understood. Here we applied two-dimensional electrophoresis (2-DE) to identify the mechanisms of mycolactone's cellular action in the L929 mouse fibroblast proteome. This reveal...

  12. Proteomic Analysis of the Action of the Mycobacterium ulcerans Toxin Mycolactone: Targeting Host Cells Cytoskeleton and Collagen

    OpenAIRE

    Gama, José B.; Ohlmeier, S.; Martins, Teresa G.; Fraga, Alexandra G.; Marques, Belém Sampaio; Carvalho, M. Alice; Proença, M. Fernanda R. P.; Silva, Manuel T.; Pedrosa, Jorge; Ludovico, Paula

    2014-01-01

    Buruli ulcer (BU) is a neglected tropical disease caused by Mycobacterium ulcerans. The tissue damage characteristic of BU lesions is known to be driven by the secretion of the potent lipidic exotoxin mycolactone. However, the molecular action of mycolactone on host cell biology mediating cytopathogenesis is not fully understood. Here we applied two-dimensional electrophoresis (2-DE) to identify the mechanisms of mycolactone's cellular action in the L929 mouse fibroblast proteome. This reveal...

  13. Whole genome analyses of marine fish pathogenic isolate, Mycobacterium sp. 012931.

    Science.gov (United States)

    Kurokawa, Satoru; Kabayama, Jun; Hwang, Seong Don; Nho, Seong Won; Hikima, Jun-ichi; Jung, Tae Sung; Kondo, Hidehiro; Hirono, Ikuo; Takeyama, Haruko; Mori, Tetsushi; Aoki, Takashi

    2014-10-01

    Mycobacterium is a genus within the order Actinomycetales that comprises of a large number of well-characterized species, several of which includes pathogens known to cause serious disease in human and animal. Here, we report the whole genome sequence of Mycobacterium sp. strain 012931 isolated from the marine fish, yellowtail (Seriola quinqueradiata). Mycobacterium sp. 012931 is a fish pathogen causing serious damage to aquaculture farms in Japan. DNA dot plot analysis showed that Mycobacterium sp. 012931 was more closely related to Mycobacterium marinum when compared across several Mycobacterium species. However, little conservation of the gene order was observed between Mycobacterium sp. 012931 and M. marinum genome. The annotated 5,464 genes of Mycobacterium sp. 012931 was classified into 26 subsystems. The insertion/deletion gene analysis shows Mycobacterium sp. 012931 had 643 unique genes that were not found in the M. marinum strains. In the virulence, disease, and defense subsystem, both insertion and deletion genes of Mycobacterium sp. 012931 were associated with the PPE gene cluster of Mycobacteria. Of seven plcB genes in Mycobacterium sp. 012931, plcB_2 and plcB_3 showed low identities with those of M. marinum strains. Therefore, Mycobacterium sp. 012931 has differences on genetic and virulence from M. marinum and may induce different interaction mechanisms between host and pathogen. PMID:24879010

  14. Molecular profiling of Mycobacterium tuberculosis identifies tuberculosinyl nucleoside products of the virulence-associated enzyme Rv3378c

    NARCIS (Netherlands)

    Layre, Emilie; Lee, Ho Jun; Young, David C.; Martinot, Amanda Jezek; Buter, Jeffrey; Minnaard, Adriaan J.; Annand, John W.; Fortune, Sarah M.; Snider, Barry B.; Matsunaga, Isamu; Rubin, Eric J.; Alber, Tom; Moody, D. Branch

    2014-01-01

    To identify lipids with roles in tuberculosis disease, we systematically compared the lipid content of virulent Mycobacterium tuberculosis with the attenuated vaccine strain Mycobacterium bovis bacillus Calmette-Guerin. Comparative lipidomics analysis identified more than 1,000 molecular differences

  15. 牛布氏杆菌和副结核分枝杆菌双重PCR检测方法的建立%Establishment of a duplex PCR assay for the differentiation of bovine Brucella and Mycobacterium paratuberculosis

    Institute of Scientific and Technical Information of China (English)

    王素华; 李孝军; 王忠才

    2012-01-01

    According to the sequences of specific Brucella gene of outer membrane protein(OMP31) and Mycobacterium paratuberculosis gene of C-2 chromosome(ISMav2) available in the GenBank,two pairs of primers were designed to establish a rapid duplex PCR assay for differentiation of Brucella and M. paratuberculosis. Results showed that the duplex PCR assay possessed a high specificity and sensitivity, with 602 bp and 246 bp amplicons amplified from Brucella and M. paratuberculosis,respectively. And none products were found from 7 other strains (Babesia bigemina, Escherichia coli, Salmonella typhimurium, Toxoplasma gondii, Streptococcus, Sphaerotilus boris, the above DNA mixture). Sensitivity of genomic DNA detection of Brucella and M. paratuberculosis were 1.92 pg and 2.51 pg respectively,with the detection limit in artificially contaminated beef as low as 6 X 104 CFU/mL and 7 X 104 CFU/mL, respectively. The duplex PCR assay was successfully constructed which will provide strongly technical support for the detection,identification and epidemiological investigations of bovine brucellosis and paratuberculosis.%为快速检测牛布氏杆菌和副结核分枝杆菌,根据GenBank中的布氏杆菌OMP31基因序列(JF918757)及副结核分枝杆菌ISMav2基因序列(AF286339)设计、合成2对特异性引物,通过对PCR条件的优化,建立了快速鉴别检测牛布氏杆菌和副结核分枝杆菌的双重PCR方法。经对建立的方法进行特异性和敏感性试验,结果表明,分别扩增出602、246bp的特异性牛布氏杆菌和副结核分枝杆菌DNA目的条带,作为对照的双芽巴贝斯虫、大肠杆菌、沙门氏菌、弓形虫、链球菌、牛放线菌的DNA及其混合物均未扩增出任何条带。牛布氏杆菌与副结核分枝杆菌的最低检测限分别为1.92和2.51pg;牛肉样品中人工污染的牛布氏杆菌和副结核分枝杆菌的检测敏感性分别为6×104和7×104 CFU/mL。该双病原检测体系的成功

  16. The draft genome of Mycobacterium aurum, a potential model organism for investigating drugs against Mycobacterium tuberculosis and Mycobacterium leprae

    KAUST Repository

    Phelan, Jody

    2015-06-04

    Mycobacterium aurum (M. aurum) is an environmental mycobacteria that has previously been used in studies of anti-mycobacterial drugs due to its fast growth rate and low pathogenicity. The M. aurum genome has been sequenced and assembled into 46 contigs, with a total length of 6.02 Mb containing 5684 annotated protein-coding genes. A phylogenetic analysis using whole genome alignments positioned M. aurum close to Mycobacterium vaccae and Mycobacterium vanbaalenii, within a clade related to fast-growing mycobacteria. Large-scale genomic rearrangements were identified by comparing the M. aurum genome to those of Mycobacterium tuberculosis and Mycobacterium leprae. M. aurum orthologous genes implicated in resistance to anti-tuberculosis drugs in M. tuberculosis were observed. The sequence identity at the DNA level varied from 68.6% for pncA (pyrazinamide drug-related) to 96.2% for rrs (streptomycin, capreomycin). We observed two homologous genes encoding the catalase-peroxidase enzyme (katG) that is associated with resistance to isoniazid. Similarly, two embB homologues were identified in the M. aurum genome. In addition to describing for the first time the genome of M. aurum, this work provides a resource to aid the use of M. aurum in studies to develop improved drugs for the pathogenic mycobacteria M. tuberculosis and M. leprae.

  17. 异烟肼耐药耻垢分枝杆菌的建立及药物敏感性的测定%Establishment and drug susceptibility test of isoniazid resistant Mycobacterium smegmatis

    Institute of Scientific and Technical Information of China (English)

    贾平平; 赵莉莉; 李晓宇; 张全; 刘振龙; 王鑫; 余利岩; 赵立勋; 岑山

    2011-01-01

    With the emergence of drug resistant tuberculosis, it is very urgent to find novel anti-tuberculosis drugs, especially novel anti-drug-resistant tuberculosis drugs. Because of the slow growth and the need to work in a biosafty environment of Mycobacterium tuberculosis, the development of evaluation of drug effect is severely impeded. In order to solve these issues, non-pathogenic fast-growing Mycobacterium smegmatis is introduced as test organism. The inhA is one of a target of isoniazid (INH) overexpression or mutation of this gene in Mycobacterium tuberculosis conferring resistant to INH. A recombinant plasmid bearing inhA was constructed and electroporated into Mycobacterium smegmatis, using shuttle expression vector pMV261. Transformants were induced to express a protein of inhA, identified by SDS-PAGE. Results show that Mycobacterium smegmatis containing inhA plasmids exhibited 100-fold or greater increased resistance to INH, but it conferred no increased resistance to others first-line anti-tuberculosis drugs. Resazurin microtiter assay plate testing of Mycobacterium smegmatis susceptibility to drugs is a rapid, simple, and inexpensive method and could decrease color background of drugs by detecting fluorescence. It will be benefit for highthroughout screening of drugs of anti-isoniazid-resistant Mycobacteria.%耐药结核病的涌现,使发展新型抗耐药结核药物变得尤为迫切.本研究选择生长快且无致病性的耻垢分枝杆菌为研究对象,探索快速评价药物抗异烟肼耐药结核分枝杆菌的能力.inhA是异烟肼的作用靶点,由于inhA的突变或者过表达可以引起结核分枝杆菌对异烟肼耐药性的产生.通过将inhA克隆入pMV261中,构建过表达inhA的耻垢分枝杆菌.结果显示,过量表达inhA的耻垢分枝杆菌对异烟肼的敏感性下降了100倍以上.建立了利用刃天青为指示剂的抗异烟肼耐药株的快速药效评价方法,可快速对药物的活性进行定性或定量

  18. Molecular cloning of gyrA and gyrB genes of mycobacterium tuberculosis: analysis of nucleotide sequence

    OpenAIRE

    Madhusudan, K.; Ramesh, V.; Nagaraja, V

    1994-01-01

    We have recently reported the cloning of gyrA and gyrB genes from Mycobacterium tuberculosis H37Ra [Curr. Science, (1994) 66, 664-667). Here, we present the complete nucleotide sequence of gyrB gene from M.tuberculosis H37Ra along with the flanking regions. The gyrA gene has been located 34 nucleotides downstream of gyrB and has been partially sequenced; both the genes seem to be transcribed from the promoter elements located upstream of gyrB coding sequence. The gyrB gene encodes a polypepti...

  19. Mycobacterium arupense, Mycobacterium heraklionense, and a Newly Proposed Species, "Mycobacterium virginiense" sp. nov., but Not Mycobacterium nonchromogenicum, as Species of the Mycobacterium terrae Complex Causing Tenosynovitis and Osteomyelitis.

    Science.gov (United States)

    Vasireddy, Ravikiran; Vasireddy, Sruthi; Brown-Elliott, Barbara A; Wengenack, Nancy L; Eke, Uzoamaka A; Benwill, Jeana L; Turenne, Christine; Wallace, Richard J

    2016-05-01

    Mycobacterium terrae complex has been recognized as a cause of tenosynovitis, with M. terrae and Mycobacterium nonchromogenicum reported as the primary etiologic pathogens. The molecular taxonomy of the M. terrae complex causing tenosynovitis has not been established despite approximately 50 previously reported cases. We evaluated 26 isolates of the M. terrae complex associated with tenosynovitis or osteomyelitis recovered between 1984 and 2014 from 13 states, including 5 isolates reported in 1991 as M. nonchromogenicum by nonmolecular methods. The isolates belonged to three validated species, one new proposed species, and two novel related strains. The majority of isolates (20/26, or 77%) belonged to two recently described species: Mycobacterium arupense (10 isolates, or 38%) and Mycobacterium heraklionense (10 isolates, or 38%). Three isolates (12%) had 100% sequence identity to each other by 16S rRNA and 99.3 to 100% identity by rpoB gene region V sequencing and represent a previously undescribed species within the M. terrae complex. There were no isolates of M. terrae or M. nonchromogenicum, including among the five isolates reported in 1991. The 26 isolates were susceptible to clarithromycin (100%), rifabutin (100%), ethambutol (92%), and sulfamethoxazole or trimethoprim-sulfamethoxazole (70%). The current study suggests that M. arupense, M. heraklionense, and a newly proposed species ("M. virginiense" sp. nov.; proposed type strain MO-233 [DSM 100883, CIP 110918]) within the M. terrae complex are the major causes of tenosynovitis and osteomyelitis in the United States, with little change over 20 years. Species identification within this complex requires sequencing methods. PMID:26962085

  20. Two-Year Monitoring of Water Samples from Dam of Iskar and the Black Sea, Bulgaria, by Molecular Analysis: Focus on Mycobacterium spp.

    Directory of Open Access Journals (Sweden)

    Stefan Panaiotov

    2015-06-01

    Full Text Available The coast of the Bulgarian Black Sea is a popular summer holiday destination. The Dam of Iskar is the largest artificial dam in Bulgaria, with a capacity of 675 million m3. It is the main source of tap water for the capital Sofia and for irrigating the surrounding valley. There is a close relationship between the quality of aquatic ecosystems and human health as many infections are waterborne. Rapid molecular methods for the analysis of highly pathogenic bacteria have been developed for monitoring quality. Mycobacterial species can be isolated from waste, surface, recreational, ground and tap waters and human pathogenicity of nontuberculose mycobacteria (NTM is well recognized. The objective of our study was to perform molecular analysis for key-pathogens, with a focus on mycobacteria, in water samples collected from the Black Sea and the Dam of Iskar. In a two year period, 38 water samples were collected—24 from the Dam of Iskar and 14 from the Black Sea coastal zone. Fifty liter water samples were concentrated by ultrafiltration. Molecular analysis for 15 pathogens, including all species of genus Mycobacterium was performed. Our results showed presence of Vibrio spp. in the Black Sea. Rotavirus A was also identified in four samples from the Dam of Iskar. Toxigenic Escherichia coli was present in both locations, based on markers for stx1 and stx2 genes. No detectable amounts of Cryptosporidium were detected in either location using immunomagnetic separation and fluorescence microscopy. Furthermore, mass spectrometry analyses did not detect key cyanobacterial toxins. On the basis of the results obtained we can conclude that for the period 2012–2014 no Mycobacterium species were present in the water samples. During the study period no cases of waterborne infections were reported.

  1. Evolution of Extensively Drug-Resistant Tuberculosis over Four Decades: Whole Genome Sequencing and Dating Analysis of Mycobacterium tuberculosis Isolates from KwaZulu-Natal.

    Directory of Open Access Journals (Sweden)

    Keira A Cohen

    2015-09-01

    Full Text Available The continued advance of antibiotic resistance threatens the treatment and control of many infectious diseases. This is exemplified by the largest global outbreak of extensively drug-resistant (XDR tuberculosis (TB identified in Tugela Ferry, KwaZulu-Natal, South Africa, in 2005 that continues today. It is unclear whether the emergence of XDR-TB in KwaZulu-Natal was due to recent inadequacies in TB control in conjunction with HIV or other factors. Understanding the origins of drug resistance in this fatal outbreak of XDR will inform the control and prevention of drug-resistant TB in other settings. In this study, we used whole genome sequencing and dating analysis to determine if XDR-TB had emerged recently or had ancient antecedents.We performed whole genome sequencing and drug susceptibility testing on 337 clinical isolates of Mycobacterium tuberculosis collected in KwaZulu-Natal from 2008 to 2013, in addition to three historical isolates, collected from patients in the same province and including an isolate from the 2005 Tugela Ferry XDR outbreak, a multidrug-resistant (MDR isolate from 1994, and a pansusceptible isolate from 1995. We utilized an array of whole genome comparative techniques to assess the relatedness among strains, to establish the order of acquisition of drug resistance mutations, including the timing of acquisitions leading to XDR-TB in the LAM4 spoligotype, and to calculate the number of independent evolutionary emergences of MDR and XDR. Our sequencing and analysis revealed a 50-member clone of XDR M. tuberculosis that was highly related to the Tugela Ferry XDR outbreak strain. We estimated that mutations conferring isoniazid and streptomycin resistance in this clone were acquired 50 y prior to the Tugela Ferry outbreak (katG S315T [isoniazid]; gidB 130 bp deletion [streptomycin]; 1957 [95% highest posterior density (HPD: 1937-1971], with the subsequent emergence of MDR and XDR occurring 20 y (rpoB L452P [rifampicin]; pnc

  2. Molecular Identification of Mycobacterium Tuberculosis and Analysis of Its Resistance to Rifampin in Sputa from Tuberculosis Suspected Patients

    International Nuclear Information System (INIS)

    An accurate identification of different species of Mycobacterium provides to allow appropriate treatment for Mycobacterium tuberculosis infection. Beside that, drug resistance of M. tuberculosis strains to rifampin is not clearly understood in contributing to the spread of tuberculosis in Indonesia. To assess the molecular mechanism of rifampin resistance, a number of clinical specimens of M. tuberculosis were analyzed their molecular nature of a part of the rpoB gene using polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP) methods. DNA's extracted from sputum samples were amplified and 32P-labeled by PCR with the specific primers and the product was analyzed their mutation conferring resistance by MDE gel electrophoresis. Of the 70 specimens tested, 57 specimens were positive for M. tuberculosis organism only, three specimens contained a mixture of M. tuberculosis and non tuberculosis mycobacteria (NTM), and 10 specimens were negative approved by Duplex PCR. Of these sixty DNA positive samples (thus the sensitivity of PCR was 85.71%), 5 (8.3%) of them suspected to contain mutations in rpoB which were associated with rifampin resistance. Even though the frequency of mutation was low, the results from our study clearly indicate that the molecular mechanism of rifampin resistance in M. tuberculosis isolates from Indonesia involves alterations in the rpoB gene. Molecular diagnosis by PCR which is fast and easy to perform is useful for early and rapid detection of TB in sputum specimen. (author)

  3. Molecular Identification of Mycobacterium tuberculosis and Analysis of Its Resistance to Rifampin in Sputa From Tuberculosis Suspected Patients

    Directory of Open Access Journals (Sweden)

    M. Syaifudin

    2010-08-01

    Full Text Available An accurate identification of different species of Mycobacterium provides to allow appropriate treatment for Mycobacterium tuberculosis infection. Beside that, drug resistance of M. tuberculosis strains to rifampin is not clearly understood in contributing to the spread of tuberculosis in Indonesia. To assess the molecular mechanism of rifampin resistance, a number of clinical specimens of M. tuberculosis were analyzed their molecular nature of a part of the rpoB gene using polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP methods. DNA’s extracted from sputum samples were amplified and 32P-labeled by PCR with the specific primers and the product was analyzed their mutation conferring resistance by MDE gel electrophoresis. Of the 70 specimens tested, 57 specimens were positive for M. tuberculosis organism only, three specimens contained a mixture of M. tuberculosis and non tuberculosis mycobacteria (NTM, and 10 specimens were negative approved by Duplex PCR. Of these sixty DNA positive samples (thus the sensitivity of PCR was 85.71%, 5 (8.3% of them suspected to contain mutations in rpoB which were associated with rifampin resistance. Even though the frequency of mutation was low, the results from our study clearly indicate that the molecular mechanism of rifampin resistance in M. tuberculosis isolates from Indonesia involves alterations in the rpoB gene. Molecular diagnosis by PCR which is fast and easy to perform is useful for early and rapid detection of TB in sputum specimen.

  4. Rapid Detection and Immune Characterization of Mycobacterium abscessus Infection in Cystic Fibrosis Patients

    OpenAIRE

    Steindor, Mathis; Nkwouano, Vanesa; MAYATEPEK, Ertan; Mackenzie, Colin R.; Schramm, Dirk; Jacobsen, Marc

    2015-01-01

    Cystic fibrosis patients are highly susceptible to infections with non-tuberculous mycobacteria. Especially Mycobacterium abscessus infections are common but reliable diagnosis is hampered by non-specific clinical symptoms and insensitive mycobacterial culture. In the present study we established novel methods for rapid detection and immune characterization of Mycobacterium abscessus infection in cystic fibrosis patients. We performed Mycobacterium abscessus specific DNA-strip- and quantitati...

  5. Rapid detection of multidrug-resistant Mycobacterium tuberculosis by use of real-time PCR and high-resolution melt analysis.

    Science.gov (United States)

    Ramirez, Melissa V; Cowart, Kelley C; Campbell, Patricia J; Morlock, Glenn P; Sikes, David; Winchell, Jonas M; Posey, James E

    2010-11-01

    The current study describes the development of a unique real-time PCR assay for the detection of mutations conferring drug resistance in Mycobacterium tuberculosis. The rifampicin resistance determinant region (RRDR) of rpoB and specific regions of katG and the inhA promoter were targeted for the detection of rifampin (RIF) and isoniazid (INH) resistance, respectively. Additionally, this assay was multiplexed to discriminate Mycobacterium tuberculosis complex (MTC) strains from nontuberculous Mycobacteria (NTM) strains by targeting the IS6110 insertion element. High-resolution melting (HRM) analysis following real-time PCR was used to identify M. tuberculosis strains containing mutations at the targeted loci, and locked nucleic acid (LNA) probes were used to enhance the detection of strains containing specific single-nucleotide polymorphism (SNP) transversion mutations. This method was used to screen 252 M. tuberculosis clinical isolates, including 154 RIF-resistant strains and 174 INH-resistant strains based on the agar proportion method of drug susceptibility testing (DST). Of the 154 RIF-resistant strains, 148 were also resistant to INH and therefore classified as multidrug resistant (MDR). The assay demonstrated sensitivity and specificity of 91% and 98%, respectively, for the detection of RIF resistance and 87% and 100% for the detection of INH resistance. Overall, this assay showed a sensitivity of 85% and a specificity of 98% for the detection of MDR strains. This method provides a rapid, robust, and inexpensive way to detect the dominant mutations known to confer MDR in M. tuberculosis strains and offers several advantages over current molecular and culture-based techniques.

  6. Animal-adapted members of the Mycobacterium tuberculosis complex endemic to the southern African subregion

    Directory of Open Access Journals (Sweden)

    Charlene Clarke

    2016-02-01

    Full Text Available Members of the Mycobacterium tuberculosis complex (MTC cause tuberculosis (TB in both animals and humans. In this article, three animal-adapted MTC strains that are endemic to the southern African subregion – that is, Mycobacterium suricattae, Mycobacterium mungi, and the dassie bacillus – are reviewed with a focus on clinical and pathological presentations, geographic distribution, genotyping methods, diagnostic tools and evolution. Moreover, factors influencing the transmission and establishment of TB pathogens in novel host populations, including ecological, immunological and genetic factors of both the host and pathogen, are discussed. The risks associated with these infections are currently unknown and further studies will be required for greater understanding of this disease in the context of the southern African ecosystem.Keywords: dassie bacillus; ecology; evolution; host jump; Mycobacterium mungi; Mycobacterium suricattae; Mycobacterium tuberculosis complex; phylogeny

  7. 利用果蝇转座子作载体构建土垢分枝杆菌变异基因文库的研究%ESTABLISHMENT OF MYCOBACTERIUM SMEGMATISB MUTAGENESIS LIBRARY WITH DROSOPHILA MAURITIANA MARINER ELEMENT VECTOR

    Institute of Scientific and Technical Information of China (English)

    刘克义; 于进芝

    2004-01-01

    目的利用果蝇mariner转座子作为载体构建土垢分枝杆菌随机变异基因文库以筛选影响脂肪酸代谢的基因. 方法将果蝇转座子mos1的末端反向重复序列置于卡那霉素抗性基因两侧,并将此种基因插入到M272B载体,用此载体转化土垢分枝杆菌,用Southern blot验证外源基因的插入.将变异的基因克隆在不同碳源的培养基上培养,筛选只在软脂酸上生长的菌落. 结果初筛了468个菌落,有5个不能在软脂酸上生长,其中2个在ICL基因上发生变异,3个在PCKA基因上发生变异.ICL为乙醛酸旁路所必须,PCKA编码糖异生的关键酶. 结论研究证明该转座子载体用于基因变异有效,变异筛选分枝杆菌中与脂肪酸代谢的基因可行.%Objective Use the Drosophila mauritiana mariner transposable element as a vector to construct Mycobacterium smegmatis random mutagenesis library in order to screen transposon mutants that affect the metabolism of fatty acids. Methods A transposon mutagenesis vector was constructed, which contains the Tn903 kanamycin resistance (KanR) gene flanked by the 5′ and 3′ inverted repeats of the D. mauritiana transposable element Mos1. M. smegmatis was transformed with the vector. The ability of transposon to integrate within the chromosome was identified using southern strategy. M. smegmatis transposon mutants were pooled and screened using different carbon sources. The clones could not grow on palmitate were selected. Results From initial screen of 468 colonies, we isolated one insertion in the isocitrate lyase gene, required for the glyoxalate shunt, and three independent insertions in the phosphoenolpyruvate carboxykinase gene, required for gluconeogenesis. Conclusion These results demonstrate the utility of this transposon mutagenesis vector, as well as the ability of our mutagenesis screen to identify mutations that affect the metabolism of fatty acids by mycobacterium.

  8. Mycobacterium ulcerans disease

    NARCIS (Netherlands)

    van der Werf, TS; Stienstra, Y; Johnson, RC; Phillips, R; Adjei, O; Fleischer, B; Wansbrough-Jones, MH; Johnson, PDR; Portaels, F; van der Graaf, WTA; Asiedu, K

    2005-01-01

    Mycobacterium ulcerans disease (Buruli ulcer) is an important health problem in several dwest African countries. It is prevalent in scattered foci around the world, predominantly in riverine areas with a humid, hot climate. We review the epidemiology, bacteriology, transmission, immunology, patholog

  9. Crystallization and preliminary X-ray diffraction analysis of prephenate dehydratase from Mycobacterium tuberculosis H37Rv

    International Nuclear Information System (INIS)

    The M. tuberculosis prephenate dehydratase was cloned, expressed, purified, crystallized by the hanging-drop vapour-diffusion method, and a complete data set collected to 3.2 Å resolution using synchrotron radiation. These results should pave the way for the three-dimensional structure determination of the enzyme and provide a framework on which to base the rational design of chemotherapeutic agents to treat tuberculosis. Tuberculosis remains the leading cause of mortality arising from a bacterial pathogen (Mycobacterium tuberculosis). There is an urgent need for the development of new antimycobacterial agents. The aromatic amino-acid pathway is essential for the survival of this pathogen and represents a target for structure-based drug design. Accordingly, the M. tuberculosis prephenate dehydratase has been cloned, expressed, purified and crystallized by the hanging-drop vapour-diffusion method using PEG 400 as a precipitant. The crystal belongs to the orthorhombic space group I222 or I212121, with unit-cell parameters a = 98.26, b = 133.22, c = 225.01 Å, and contains four molecules in the asymmetric unit. A complete data set was collected to 3.2 Å resolution using a synchrotron-radiation source

  10. Strategic analysis on establishing a natural gas trading hub in China

    Directory of Open Access Journals (Sweden)

    Xiaoguang Tong

    2014-12-01

    Full Text Available Since 2010, the LNG importing price premium in the Asia–Pacific markets has become increasingly high, generating great effects on the economic development in China. In addition, the natural gas dependence degree is expanding continuously, making it extremely urgent to establish a natural gas trading hub in China, with the aim to ensure national energy security, to gain the pricing power, and to build the regional benchmark prices. Through a comparative analysis of internal strength/weakness and external competitiveness, we concluded that with intensively-issued supporting policies on the natural gas sector, the initiation of spot and futures markets, the rapid growth of gas production and highly-improved infrastructures, as well as Shanghai's advantageous location, China has more advantages in establishing an Asian Natural Gas Trading Hub than other counties like Singapore, Japan and Malaysia. Moreover, based on the SWOT (strength, weakness, opportunity and threat and the marketization process analysis, the following strategies were presented: to impel the establishment of a natural gas trading hub depending on the gas supply condition, to follow the policies to complete the gas storage system, to form regional communities by taking comparative advantages, and to reinforce the marketization reform and regulation system establishment with foreign experiences for reference. This study rationalized the necessity and practicality of establishing a natural gas trading hub in China and will help China make a proper decision and find a periodical strategic path in this sector.

  11. Secretome profile analysis of hypervirulent Mycobacterium tuberculosis CPT31 reveals increased production of EsxB and proteins involved in adaptation to intracellular lifestyle.

    Science.gov (United States)

    Vargas-Romero, Fernado; Guitierrez-Najera, Nora; Mendoza-Hernández, Guillermo; Ortega-Bernal, Daniel; Hernández-Pando, Rogelio; Castañón-Arreola, Mauricio

    2016-03-01

    Epidemiological information and animal models have shown various Mycobacterium tuberculosis phenotypes ranging from hyper- to hypovirulent forms. Recent genomic and proteomic studies suggest that the outcome of infection depends on the M. tuberculosis fitness, which is a direct consequence of its phenotype. However, little is known about the molecular and cellular mechanisms used by mycobacteria to survive, replicate and persist during infection. The aim of this study was to perform a comprehensive proteomic analysis of culture filtrate from hypo- (CPT23) and hypervirulent (CPT31) M. tuberculosis isolates. Using two-dimensional electrophoresis we observed that 70 proteins were unique, or more abundant in culture filtrate of CPT31, and 15 of these were identified by mass spectrometry. Our analysis of protein expression showed that most of the proteins identified are involved in lipid metabolism (FadA3, FbpB and EchA3), detoxification and adaptation (GroEL2, SodB and HspX) and cell wall processes (LprA, Tig and EsxB). These results suggest that overrepresented proteins in M. tuberculosis CPT31 secretome could facilitate mycobacterial infection and persistence.

  12. [Methods of detector response function establishment in X-ray fluorescence spectra analysis].

    Science.gov (United States)

    Li, Zhe; Tuo, Xian-Guo; Yang, Jian-Bo; Liu, Ming-Zhe; Cheng, Yi; Wang, Lei; Zhou, Jian-Bin

    2012-11-01

    During the measurement and analysis process of X-ray fluorescence spectra, it is very helpful to improve the analyze speed, accuracy and automaticity of X-ray fluorescence spectra analysis by establishing detector response function(DRF), which represents the shape of full energy peak and can provide former basic data for subsequent X-ray analysis technique. For the theory and model of semiconductor DRF in X-ray energy spectrum measurements, methods of three typical detector response function model establishment, key parameters of full energy peak standard deviation and Fano factor calculation, etc. are discussed, and meanwhile, the summarization and contrast of existing studies are shown in this paper. Finally, the suggestion for modeling methods of DRF in X-ray fluorescence spectra measurements is provided. PMID:23387190

  13. Population density and efficiency in energy consumption: An empirical analysis of service establishments

    International Nuclear Information System (INIS)

    This study, using novel establishment-level microdata from the Energy Consumption Statistics, empirically analyzes the effect of urban density on energy intensity in the service sector. According to the analysis, the efficiency of energy consumption in service establishments is higher for densely populated cities. Quantitatively, after controlling for differences among industries, energy efficiency increases by approximately 12% when the density in a municipality population doubles. This result suggests that, given a structural transformation toward the service economy, deregulation of excessive restrictions hindering urban agglomeration, and investment in infrastructure in city centers would contribute to environmentally friendly economic growth.

  14. Proteomic analysis of the action of the Mycobacterium ulcerans toxin mycolactone: targeting host cells cytoskeleton and collagen.

    Directory of Open Access Journals (Sweden)

    José B Gama

    2014-08-01

    Full Text Available Buruli ulcer (BU is a neglected tropical disease caused by Mycobacterium ulcerans. The tissue damage characteristic of BU lesions is known to be driven by the secretion of the potent lipidic exotoxin mycolactone. However, the molecular action of mycolactone on host cell biology mediating cytopathogenesis is not fully understood. Here we applied two-dimensional electrophoresis (2-DE to identify the mechanisms of mycolactone's cellular action in the L929 mouse fibroblast proteome. This revealed 20 changed spots corresponding to 18 proteins which were clustered mainly into cytoskeleton-related proteins (Dync1i2, Cfl1, Crmp2, Actg1, Stmn1 and collagen biosynthesis enzymes (Plod1, Plod3, P4ha1. In line with cytoskeleton conformational disarrangements that are observed by immunofluorescence, we found several regulators and constituents of both actin- and tubulin-cytoskeleton affected upon exposure to the toxin, providing a novel molecular basis for the effect of mycolactone. Consistent with these cytoskeleton-related alterations, accumulation of autophagosomes as well as an increased protein ubiquitination were observed in mycolactone-treated cells. In vivo analyses in a BU mouse model revealed mycolactone-dependent structural changes in collagen upon infection with M. ulcerans, associated with the reduction of dermal collagen content, which is in line with our proteomic finding of mycolactone-induced down-regulation of several collagen biosynthesis enzymes. Our results unveil the mechanisms of mycolactone-induced molecular cytopathogenesis on exposed host cells, with the toxin compromising cell structure and homeostasis by inducing cytoskeleton alterations, as well as disrupting tissue structure, by impairing the extracellular matrix biosynthesis.

  15. Proteomic analysis of the action of the Mycobacterium ulcerans toxin mycolactone: targeting host cells cytoskeleton and collagen.

    Science.gov (United States)

    Gama, José B; Ohlmeier, Steffen; Martins, Teresa G; Fraga, Alexandra G; Sampaio-Marques, Belém; Carvalho, Maria A; Proença, Fernanda; Silva, Manuel T; Pedrosa, Jorge; Ludovico, Paula

    2014-08-01

    Buruli ulcer (BU) is a neglected tropical disease caused by Mycobacterium ulcerans. The tissue damage characteristic of BU lesions is known to be driven by the secretion of the potent lipidic exotoxin mycolactone. However, the molecular action of mycolactone on host cell biology mediating cytopathogenesis is not fully understood. Here we applied two-dimensional electrophoresis (2-DE) to identify the mechanisms of mycolactone's cellular action in the L929 mouse fibroblast proteome. This revealed 20 changed spots corresponding to 18 proteins which were clustered mainly into cytoskeleton-related proteins (Dync1i2, Cfl1, Crmp2, Actg1, Stmn1) and collagen biosynthesis enzymes (Plod1, Plod3, P4ha1). In line with cytoskeleton conformational disarrangements that are observed by immunofluorescence, we found several regulators and constituents of both actin- and tubulin-cytoskeleton affected upon exposure to the toxin, providing a novel molecular basis for the effect of mycolactone. Consistent with these cytoskeleton-related alterations, accumulation of autophagosomes as well as an increased protein ubiquitination were observed in mycolactone-treated cells. In vivo analyses in a BU mouse model revealed mycolactone-dependent structural changes in collagen upon infection with M. ulcerans, associated with the reduction of dermal collagen content, which is in line with our proteomic finding of mycolactone-induced down-regulation of several collagen biosynthesis enzymes. Our results unveil the mechanisms of mycolactone-induced molecular cytopathogenesis on exposed host cells, with the toxin compromising cell structure and homeostasis by inducing cytoskeleton alterations, as well as disrupting tissue structure, by impairing the extracellular matrix biosynthesis. PMID:25101965

  16. Comparative Proteomic Analysis of Aminoglycosides Resistant and Susceptible Mycobacterium tuberculosis Clinical Isolates for Exploring Potential Drug Targets.

    Directory of Open Access Journals (Sweden)

    Divakar Sharma

    Full Text Available Aminoglycosides, amikacin (AK and kanamycin (KM are second line anti-tuberculosis drugs used to treat tuberculosis (TB and resistance to them affects the treatment. Membrane and membrane associated proteins have an anticipated role in biological processes and pathogenesis and are potential targets for the development of new diagnostics/vaccine/therapeutics. In this study we compared membrane and membrane associated proteins of AK and KM resistant and susceptible Mycobacterium tuberculosis isolates by 2DE coupled with MALDI-TOF/TOF-MS and bioinformatic tools. Twelve proteins were found to have increased intensities (PDQuest Advanced Software in resistant isolates and were identified as ATP synthase subunit alpha (Rv1308, Trigger factor (Rv2462c, Dihydrolipoyl dehydrogenase (Rv0462, Elongation factor Tu (Rv0685, Transcriptional regulator MoxR1(Rv1479, Universal stress protein (Rv2005c, 35kDa hypothetical protein (Rv2744c, Proteasome subunit alpha (Rv2109c, Putative short-chain type dehydrogenase/reductase (Rv0148, Bacterioferritin (Rv1876, Ferritin (Rv3841 and Alpha-crystallin/HspX (Rv2031c. Among these Rv2005c, Rv2744c and Rv0148 are proteins with unknown functions. Docking showed that both drugs bind to the conserved domain (Usp, PspA and SDR domain of these hypothetical proteins and GPS-PUP predicted potential pupylation sites within them. Increased intensities of these proteins and proteasome subunit alpha might not only be neutralized/modulated the drug molecules but also involved in protein turnover to overcome the AK and KM resistance. Besides that Rv1876, Rv3841 and Rv0685 were found to be associated with iron regulation signifying the role of iron in resistance. Further research is needed to explore how these potential protein targets contribute to resistance of AK and KM.

  17. Comparative Proteomic Analysis of Aminoglycosides Resistant and Susceptible Mycobacterium tuberculosis Clinical Isolates for Exploring Potential Drug Targets

    Science.gov (United States)

    Sharma, Divakar; Kumar, Bhavnesh; Lata, Manju; Joshi, Beenu; Venkatesan, Krishnamurthy; Shukla, Sangeeta; Bisht, Deepa

    2015-01-01

    Aminoglycosides, amikacin (AK) and kanamycin (KM) are second line anti-tuberculosis drugs used to treat tuberculosis (TB) and resistance to them affects the treatment. Membrane and membrane associated proteins have an anticipated role in biological processes and pathogenesis and are potential targets for the development of new diagnostics/vaccine/therapeutics. In this study we compared membrane and membrane associated proteins of AK and KM resistant and susceptible Mycobacterium tuberculosis isolates by 2DE coupled with MALDI-TOF/TOF-MS and bioinformatic tools. Twelve proteins were found to have increased intensities (PDQuest Advanced Software) in resistant isolates and were identified as ATP synthase subunit alpha (Rv1308), Trigger factor (Rv2462c), Dihydrolipoyl dehydrogenase (Rv0462), Elongation factor Tu (Rv0685), Transcriptional regulator MoxR1(Rv1479), Universal stress protein (Rv2005c), 35kDa hypothetical protein (Rv2744c), Proteasome subunit alpha (Rv2109c), Putative short-chain type dehydrogenase/reductase (Rv0148), Bacterioferritin (Rv1876), Ferritin (Rv3841) and Alpha-crystallin/HspX (Rv2031c). Among these Rv2005c, Rv2744c and Rv0148 are proteins with unknown functions. Docking showed that both drugs bind to the conserved domain (Usp, PspA and SDR domain) of these hypothetical proteins and GPS-PUP predicted potential pupylation sites within them. Increased intensities of these proteins and proteasome subunit alpha might not only be neutralized/modulated the drug molecules but also involved in protein turnover to overcome the AK and KM resistance. Besides that Rv1876, Rv3841 and Rv0685 were found to be associated with iron regulation signifying the role of iron in resistance. Further research is needed to explore how these potential protein targets contribute to resistance of AK and KM. PMID:26436944

  18. Comparative Proteomic Analysis of Aminoglycosides Resistant and Susceptible Mycobacterium tuberculosis Clinical Isolates for Exploring Potential Drug Targets.

    Science.gov (United States)

    Sharma, Divakar; Kumar, Bhavnesh; Lata, Manju; Joshi, Beenu; Venkatesan, Krishnamurthy; Shukla, Sangeeta; Bisht, Deepa

    2015-01-01

    Aminoglycosides, amikacin (AK) and kanamycin (KM) are second line anti-tuberculosis drugs used to treat tuberculosis (TB) and resistance to them affects the treatment. Membrane and membrane associated proteins have an anticipated role in biological processes and pathogenesis and are potential targets for the development of new diagnostics/vaccine/therapeutics. In this study we compared membrane and membrane associated proteins of AK and KM resistant and susceptible Mycobacterium tuberculosis isolates by 2DE coupled with MALDI-TOF/TOF-MS and bioinformatic tools. Twelve proteins were found to have increased intensities (PDQuest Advanced Software) in resistant isolates and were identified as ATP synthase subunit alpha (Rv1308), Trigger factor (Rv2462c), Dihydrolipoyl dehydrogenase (Rv0462), Elongation factor Tu (Rv0685), Transcriptional regulator MoxR1(Rv1479), Universal stress protein (Rv2005c), 35kDa hypothetical protein (Rv2744c), Proteasome subunit alpha (Rv2109c), Putative short-chain type dehydrogenase/reductase (Rv0148), Bacterioferritin (Rv1876), Ferritin (Rv3841) and Alpha-crystallin/HspX (Rv2031c). Among these Rv2005c, Rv2744c and Rv0148 are proteins with unknown functions. Docking showed that both drugs bind to the conserved domain (Usp, PspA and SDR domain) of these hypothetical proteins and GPS-PUP predicted potential pupylation sites within them. Increased intensities of these proteins and proteasome subunit alpha might not only be neutralized/modulated the drug molecules but also involved in protein turnover to overcome the AK and KM resistance. Besides that Rv1876, Rv3841 and Rv0685 were found to be associated with iron regulation signifying the role of iron in resistance. Further research is needed to explore how these potential protein targets contribute to resistance of AK and KM.

  19. Biochemical analysis of the NAD+-dependent malate dehydrogenase, a substrate of several serine/threonine protein kinases of Mycobacterium tuberculosis.

    Directory of Open Access Journals (Sweden)

    Xiao Ming Wang

    Full Text Available PknD is one of the eleven eukaryotic-like serine/threonine protein kinases (STPKs of Mycobacterium tuberculosis (Mtb. In vitro phosphorylation assays with the active recombinant PknD showed that the intracellular protein NAD+-dependent malate dehydrogenase (MDH is a substrate of this kinase. MDH, an energy-supplying enzyme, catalyzes the interconversion of malate and oxaloacetate and plays crucial roles in several metabolic pathways including the citric acid cycle. The phosphorylation site was identified on threonine residues and the phosphorylation inhibited the MDH activity. In vitro, the recombinant MDH could also be phosphorylated by at least five other STPKs, PknA, PknE, PknH, PknJ, and PknG. Immunoprecipitation analysis revealed that MDH was hyperphosphorylated in the bacteria at the beginning of the stationary and under oxygen-limited conditions by STPKs other than PknD. On the contrary, when PknD-deficient mutant mycobacteria were grown in a phosphate-depleted medium, MDH was not detectably phosphorylated. These results suggest that although the MDH is a substrate of several mycobacterial STPKs, the activity of these kinases can depend on the environment, as we identified PknD as a key element in the MDH phosphorylation assay under phosphate-poor conditions.

  20. Proteomics Analysis of Three Different Strains of Mycobacterium tuberculosis under In vitro Hypoxia and Evaluation of Hypoxia Associated Antigen's Specific Memory T Cells in Healthy Household Contacts.

    Science.gov (United States)

    Devasundaram, Santhi; Gopalan, Akilandeswari; Das, Sulochana D; Raja, Alamelu

    2016-01-01

    In vitro mimicking conditions are thought to reflect the environment experienced by Mycobacterium tuberculosis inside the host granuloma. The majority of in vitro dormancy experimental models use laboratory-adapted strains H37Rv or Erdman instead of prevalent clinical strains involved during disease outbreaks. Thus, we included the most prevalent clinical strains (S7 and S10) of M. tuberculosis from south India in addition to H37Rv for our in vitro oxygen depletion (hypoxia) experimental model. Cytosolic proteins were prepared from hypoxic cultures, resolved by two-dimensional electrophoresis and protein spots were characterized by mass spectrometry. In total, 49 spots were characterized as over-expressed or newly emergent between the three strains. Two antigens (ESAT-6, Lpd) out of the 49 characterized spots were readily available in recombinant form in our lab. Hence, these two genes were overexpressed, purified and used for in vitro stimulation of whole blood collected from healthy household contacts (HHC) and active pulmonary tuberculosis patients (PTB). Multicolor flow cytometry analysis showed high levels of antigen specific CD4(+) central memory T cells in the circulation of HHC compared to PTB (p hypoxia in most prevalent clinical strains would indicate possible potential immunogens. In vitro hypoxia experiments with most prevalent clinical strains would also elucidate the probable true representative antigens involved in adaptive mechanisms. PMID:27667981

  1. Inactivation of Mycobacterium paratuberculosis and Mycobacterium tuberculosis in fresh soft cheese by gamma radiation

    Energy Technology Data Exchange (ETDEWEB)

    Badr, Hesham M., E-mail: heshambadr_aea@yahoo.co.uk [Atomic Energy Authority, Nuclear Research Center, Abou Zaabal, P.O. Box 13759 Cairo (Egypt)

    2011-11-15

    The effectiveness of gamma irradiation on the inactivation of Mycobacterium paratuberculosis, Mycobacterium bovis and Mycobacterium tuberculosis in fresh soft cheese that prepared from artificially inoculated milk samples was studied. Irradiation at dose of 2 kGy was sufficient for the complete inactivation of these mycobacteria as they were not detected in the treated samples during storage at 4{+-}1 {sup o}C for 15 days. Moreover, irradiation of cheese samples, that were prepared from un-inoculated milk, at this effective dose had no significant effects on their gross composition and contents from riboflavin, niacin and pantothenic acid, while significant decreases in vitamin A and thiamin were observed. In addition, irradiation of cheese samples had no significant effects on their pH and nitrogen fractions contents, except for the contents of ammonia, which showed a slight, but significant, increases due to irradiation. The analysis of cheese fats indicated that irradiation treatment induced significant increase in their oxidation parameters and contents from free fatty acids; however, the observed increases were relatively low. On the other hand, irradiation of cheese samples induced no significant alterations on their sensory properties. Thus, irradiation dose of 2 kGy can be effectively applied to ensure the safety of soft cheese with regards to these harmful mycobacteria. - Highlights: > We examined the effectiveness of gamma irradiation on inactivation of Mycobacterium paratuberculosis, Mycobacterium bovis and Mycobacterium tuberculosis in fresh soft cheese. > Irradiation at dose of 2 kGy was sufficient for complete inactivation of these mycobacteria. > Irradiation of cheese samples induced no significant alterations on their sensory properties.

  2. Establishment and application of the cyclic neutron activation analysis method on mini-reactor

    International Nuclear Information System (INIS)

    Background: Instrument neutron activation analysis is a nondestructive analytic method. Some elements after irradiation produce short half-life radio-nuclides (<60 s), and others produce both long-lived and short-lived nuclides. For these short-lived nuclides, the single measurement has big error. Purpose: In order to reduce the error, cyclic neutron activation analysis can be used to improve the sensitivity. Methods: A device was designed to be connected to the sample transporter, detector and irradiation pipeline in the reactor, which can automatically control the irradiation time and counting time. According to the nuclear parameters of certain elements, irradiation time and counting time and cycle times were determined by experiment. Cyclic activation analysis method was established at the mini-reactor. Results: This paper studied cyclic activation analysis conditions of 17 kinds of element, and applied to the determination of actual samples. Cyclic epithermal neutron activation analysis (CENAA) method was discussed too. By the analysis of national standard reference materials, the reliability of this method was confirmed. Conclusion: Cyclic neutron activation analysis (CNAA) is an effective analytic method for only short life nuclide elements. For both short and long lived nuclides of elements, the cyclic activation analysis method can make the analytical cycle shorten, from a few days or several weeks to within a few minutes. Cyclic activation analysis has the advantages of high sensitivity, and its precision and accuracy are better than single short irradiation activation analysis. (authors)

  3. Isolation of Mycobacterium kumamotonense from a patient with pulmonary infection and latent tuberculosis.

    Science.gov (United States)

    Kontos, Fanourios; Mavromanolakis, Dimitrios Nikitas; Zande, Marina Chari; Gitti, Zoe Georgios

    2016-01-01

    Mycobacterium kumamotonense is a novel, slow-growing non-chromogenic nontuberculous mycobacterium, which belongs to Mycobacterium terrae complex. We report, for the first time in Greece, the isolation of M. kumamotonense from an immunocompetent patient with pulmonary infection and latent tuberculosis. M. kumamotonense was identified by sequencing analysis of 16S rDNA and 65-kDa heat shock protein genes while by commercial molecular assays it was misidentified as Mycobacterium celatum. Antibiotic susceptibility testing was performed by the reference broth microdilution method. The strain was susceptible to amikacin, clarithromycin, rifampin, ciprofloxacin, moxifloxacin, rifabutin, ethambutol and linezolid. PMID:27080783

  4. Establishment of Grain Farmers' Supply Response Model and Empirical Analysis under Minimum Grain Purchase Price Policy

    OpenAIRE

    Shuang ZHANG

    2012-01-01

    Based on farmers' supply behavior theory and price expectations theory, this paper establishes grain farmers' supply response model of two major grain varieties (early indica rice and mixed wheat) in the major producing areas, to test whether the minimum grain purchase price policy can have price-oriented effect on grain production and supply in the major producing areas. Empirical analysis shows that the minimum purchase price published annually by the government has significant positive imp...

  5. Analysis on Systematic Water Scarcity Based on Establishment of Water Scarcity Classification System

    Institute of Scientific and Technical Information of China (English)

    2011-01-01

    It would be very helpful for making countermeasures against complex water scarcity by analysis on systematic water scarcity.Based on the previous researches on water scarcity classification,a classification system of water scarcity was established according to contributing factors,which comprises three water scarcity categories caused by anthropic factors,natural factors and mixed factors respectively.Accordingly,the concept of systematic water scarcity was proposed,which can be defined as one type of water...

  6. Evolution of Mycobacterium tuberculosis.

    Science.gov (United States)

    Behr, Marcel A

    2013-01-01

    Genomic studies have provided a refined understanding of the genetic diversity within the Mycobacterium genus, and more specifically within Mycobacterium tuberculosis. These results have informed a new perspective on the macro- and micro-evolution of the tubercle bacillus. In the first step, a M. kansasii-like opportunistic pathogen acquired new genes, through horizontal gene transfer, that enabled it to better exploit an intracellular niche and ultimately evolve into a professional pathogen. In the second step, different subspecies and strains of the M. tuberculosis complex emerged through mutation and deletion of unnecessary DNA. Understanding the differences between M. tuberculosis and related less pathogenic mycobacteria is expected to reveal key bacterial virulence mechanisms and provide opportunities to understand host resistance to mycobacterial infection. Understanding differences within the M. tuberculosis complex and the evolutionary forces shaping these differences is important for investigating the basis of its success as both a symbiont and a pathogen.

  7. Antipyretic therapy in critically ill patients with established sepsis: a trial sequential analysis.

    Directory of Open Access Journals (Sweden)

    Zhongheng Zhang

    Full Text Available antipyretic therapy for patients with sepsis has long been debated. The present study aimed to explore the beneficial effect of antipyretic therapy for ICU patients with sepsis.systematic review and trial sequential analysis of randomized controlled trials.Pubmed, Scopus, EBSCO and EMBASE were searched from inception to August 5, 2014.Mortality was dichotomized as binary outcome variable and odds ratio (OR was chosen to be the summary statistic. Pooled OR was calculated by using DerSimonian and Laird method. Statistical heterogeneity was assessed by using the statistic I2. Trial sequential analysis was performed to account for the small number of trials and patients.A total of 6 randomized controlled trials including 819 patients were included into final analysis. Overall, there was no beneficial effect of antipyretic therapy on mortality risk in patients with established sepsis (OR: 1.02, 95% CI: 0.50-2.05. The required information size (IS was 2582 and our analysis has not yet reached half of the IS. The Z-curve did not cross the O'Brien-Fleming α-spending boundary or reach the futility, indicating that the non-significant result was probably due to lack of statistical power.our study fails to identify any beneficial effect of antipyretic therapy on ICU patients with established diagnosis of sepsis. Due to limited number of total participants, more studies are needed to make a conclusive and reliable analysis.

  8. Establishment of a proximate analysis of coal based on the use of a thermobalance

    Energy Technology Data Exchange (ETDEWEB)

    Matsumoto, S.; Watanuki, O.

    1986-01-01

    Studies are reported aimed at the establishment of a new method for the proximate analysis of coal. The results obtained are expected to clear the way for the introduction of this thermogravimetric method. Firstly, in order to achieve a repeatability as good as that of the JIS method and such that operating problems do not result at power plants, measurement procedures were standardized on the basis of an analysis of the factors affecting repeatability. Secondly, a regression analysis was carried out to check the correspondence between the JIS method and the thermogravimetric method over a practical operating range and for all the types of coal presently received at power plants. This analysis confirmed the linearity of the relations between the two sets of results. 2 references, 4 figures.

  9. Prevalence and occurrence rate of Mycobacterium tuberculosis Haarlem family multi-drug resistant in the worldwide population: A systematic review and meta-analysis

    Directory of Open Access Journals (Sweden)

    Rashid Ramazanzadeh

    2015-01-01

    Full Text Available Background: Transmission of Mycobacterium tuberculosis (M. tuberculosis can occur in different ways. Furthermore, drug resistant in M. tuberculosis family is a major problem that creates obstacles in treatment and control of tuberculosis (TB in the world. One of the most prevalent families of M. tuberculosis is Haarlem, and it is associated with drug resistant. Our objectives of this study were to determine the prevalence and occurrence rate of M. tuberculosis Haarlem family multi-drug resistant (MDR in the worldwide using meta-analysis based on a systematic review that performed on published articles. Materials and Methods: Data sources of this study were 78 original articles (2002-2012 that were published in the literatures in several databases including PubMed, Science Direct, Google Scholar, Biological abstracts, ISI web of knowledge and IranMedex. The articles were systematically reviewed for prevalence and rate of MDR. Data were analyzed using meta-analysis and random effects models with the software package Meta R, Version 2.13 (P < 0.10. Results: Final analysis included 28601 persons in 78 articles. The highest and lowest occurrence rate of Haarlem family in M. tuberculosis was in Hungary in 2006 (66.20% with negative MDR-TB and in China in 2010 (0.8%, respectively. From 2002 to 2012, the lowest rate of prevalence was in 2010, and the highest prevalence rate was in 2012. Also 1.076% were positive for MDR and 9.22% were negative (confidence interval: 95%.0020. Conclusion: Many articles and studies are performed in this field globally, and we only chose some of them. Further studies are needed to be done in this field. Our study showed that M. tuberculosis Haarlem family is prevalent in European countries. According to the presence of MDR that was seen in our results, effective control programs are needed to control the spread of drug-resistant strains, especially Haarlem family.

  10. EFFECT OF SOME MEDICINAL PLANTS ON GROWTH OF MYCOBACTERIUM TUBERCULOSIS, MULTI DRUG RESISTANT MYCOBACTERIUM TUBERCULOSIS AND MYCOBACTERIUM OTHER THAN TUBERCULOSIS

    OpenAIRE

    Prashant Shukla; Ajay Sharma

    2013-01-01

    Six plants of medicinal uses were tried for their inhibitory effect on Mycobacterium tuberculosis (MTB), multi drug resistant Mycobacterium tuberculosis (MDR MTB) and Mycobacterium other than tuberculosis (MOTT). MTB, MDR MTB and MOTT were cultured in 12B medium vials for Bacterc 460 TB system and incubated at 37˚C. The vials were read in Bacterc 460 TB system. Garlic, Ocimum sanctum, onion and neem showed effectiveness towards Mycobacterium tuberculosis and multi drug resistant Mycobacterium...

  11. Analysis of mutations in the gyrA and gyrB genes and their association with the resistance of Mycobacterium tuberculosis to levofloxacin, moxifloxacin and gatifloxacin.

    Science.gov (United States)

    Nosova, Elena Yu; Bukatina, Anastasia A; Isaeva, Yulia D; Makarova, Marina V; Galkina, Ksenia Yu; Moroz, Arkadyi M

    2013-01-01

    The purpose of the present study was to analyse mutations in the gyrA and gyrB genes of Mycobacterium tuberculosis and define the possible correlation between these mutations and resistance to levofloxacin (LVX), moxifloxacin (MFX) and gatifloxacin (GAT), based on their MICs. One hundred and forty-two M. tuberculosis clinical isolates were collected from pulmonary tuberculosis patients in the Moscow region. All M. tuberculosis strains were tested for drug susceptibility to rifampicin and isoniazid using the BACTEC MGIT 960 System and to ofloxacin (OFX) using the absolute concentration method on solid Lowenstein-Jensen slants. All in all, 68 strains were selected at random (38 strains were resistant and 30 were susceptible to OFX) for further analysis using the TB-BIOCHIP-2 test system and DNA sequence analysis. The MICs of LVX, MFX and GAT for selected strains were determined using the BACTEC MGIT 960 System. Mutations in the gyrA gene were observed in 36 out of 38 (94.7 %) OFX-resistant M. tuberculosis strains. Asn538Asp and Asp500His substitutions in the gyrB gene only were found in two (5.3 %) strains. Twenty-nine out of 30 OFX-sensitive M. tuberculosis strains had no mutations in either gene. One (3.3 %) OFX-sensitive M. tuberculosis strain carried an Arg485His substitution in gyrB. The results of our investigation showed that there is no clear correlation between the type of mutation in the genes gyrA and gyrB, and the MIC levels of LVX, MFX and GAT for resistant strains. Mutations in gyrA and Asn538Asp, and Asp500His substitutions in gyrB were associated with cross-resistance of M. tuberculosis to fluoroquinolones. The substitution Arg485His in gyrB does not confer resistance to LVX, MFX and GAT in M. tuberculosis.

  12. Functional analysis of the newly established plants induced by nesting gulls on Riou archipelago (Marseille, France)

    Science.gov (United States)

    Vidal, Eric; Médail, Frédéric; Tatoni, Thierry; Vidal, Patrick; Roche, Philip

    1998-06-01

    The recent population explosion of Yellow-legged gulls ( Larus cachinnans), nesting on the Riou archipelago, off Marseille (France), has perturbed the flora and the vegetation of this site. The present study consists of a functional approach to the newly established plant species through an analysis of some of their vital attributes. Small islets appear to be more affected by floristic turnover than larger islands. In some cases, more than 50 % of the current flora was not present 35 years ago. The newly established taxa show special adaptations to the severe ecological pressure induced by gull colonies and to the characteristics of the Mediterranean climate. These plant species are mostly therophytes or hemicryptophytes and have a ruderal or a stress-tolerant strategy. Gulls' contribution to propagule dispersal from the continent appears to be very slight, dispersal by wind being the prevalent mode.

  13. Mycobacterium abscessus skin infection after tattooing--Case report.

    Science.gov (United States)

    Sousa, Pétra Pereira de; Cruz, Rossilene Conceição da Silva; Schettini, Antonio Pedro Mendes; Westphal, Danielle Cristine

    2015-01-01

    Mycobacterium abscessus is a rapidly growing mycobacterium that has been affecting people undergoing invasive procedures, such as videosurgery and mesotherapy. This bacterium has global distribution, being found in numerous niches. The frequency of published reports of infection by rapidly growing mycobacteria associated with tattooing procedures has increased in recent years. However, in Brazil there were no case reports of M. abscessus after tattooing in the literature until now. In this paper, we describe the case of a patient with a nine-month history of lesion on a tattoo site. The diagnosis of infection with Mycobacterium abscessus was established by correlation between dermatological and histopathological aspects, culture and molecular biology techniques. The patient had significant improvement of symptoms with the use of clarithromycin monotherapy. PMID:26560222

  14. Establishment of joint application system of safety analysis codes between Korea and Vietnam

    International Nuclear Information System (INIS)

    The following KAERI-VAEI collaboration works have been performed during the 2 year project ('09.4∼'11.4). 1) On the job training of Vietnam code users(1st training for 4 VAEI staff-3 months. 2nd training for 3 VAEI staff- 3 month), 2) Lecture of nuclear safety analysis (30 hrs basic course and 30 hrs advanced course), 3) Review of safety analysis method (IAEA safety concept and requirements), 4) Collaborative assessment of safety analysis code MARS (13 conceptual problem, 2 separate effect test problem, 1 integral effect test problem), 5) Input deck preparation of standard PWR (Preparation of APR1400 input deck and safety analysis of DBA). VAEI staffs have been familiarized to Korean PWR safety assessment technology through the collaboration assessment work using a computer code developed in Korea. The lectures for Vietnamese research will be contributed to the utilization and cultivation of Korean safety technology. The collaborated assessment works will be used for the establishment of MARS based safety analysis system which is independent from US safety assessment system

  15. 21 CFR 866.3370 - Mycobacterium tuberculosis immunofluorescent reagents.

    Science.gov (United States)

    2010-04-01

    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Mycobacterium tuberculosis immunofluorescent... § 866.3370 Mycobacterium tuberculosis immunofluorescent reagents. (a) Identification. Mycobacterium... used to identify Mycobacterium tuberculosis directly from clinical specimens. The identification...

  16. Septic arthritis caused by Mycobacterium marinum.

    Science.gov (United States)

    Riera, Jaume; Conesa, Xavier; Pisa, Jose; Moreno, Josefa; Siles, Eduard; Novell, Josep

    2016-01-01

    The incidence of infection by Mycobacterium marinum is rising, mainly due to the increasing popularity of home aquariums. The infection typically manifests as skin lesions, with septic arthritis being a rare presentation form. The disease is difficult to diagnose even when there is a high clinical suspicion, as culture in specific media may not yield positive findings. Thus, establishment of appropriate treatment is often delayed. Synovectomy, capsular thinning, and joint drainage together with prolonged, combined antibiotic therapy may be needed to cure the infection.

  17. Technical analysis for the establishment of the test and design requirement of FTL

    Energy Technology Data Exchange (ETDEWEB)

    Sim, B.S.; Wu, J.S.; Kim, H.R. [and others

    1999-03-01

    A task force team was made up to establish the design and test requirements of the fuel test loop(FTL) which will be installed in HANARO. The activities of the task force team include the review of the performance of the FTL, the establishment of user requirement, the review of interface between HANARO and FTL, the reevaluation of design concept and finally the provision of the viable design option through the impact analysis on design change. Based on the review, the task force team has finalized the design concept of the in-pile test section(IPS) which accommodates a maximum number of 7 fuel pins. Following the modification of the IPS, we have identified the required design modification of the out-file system(OPS) to reduce the investment cost FTL facility while maintaining the performance and not jeopardizing the safety of system. The result of the analysis will be used for the designchange of both IPS and OPS system of FTL and for the future works of the project. (author). 30 tabs., 104 figs.

  18. Establishment of a clean laboratory for ultra trace analysis of nuclear materials in safeguards environmental samples

    Energy Technology Data Exchange (ETDEWEB)

    Hanzawa, Yukiko; Magara, Masaaki; Watanabe, Kazuo [Japan Atomic Energy Research Inst., Tokai, Ibaraki (Japan). Tokai Research Establishment] (and others)

    2003-01-01

    The Japan Atomic Energy Research Institute has established a cleanroom facility with cleanliness of ISO Class 5: the Clean Laboratory for Environmental Analysis and Research (CLEAR). It was designed to be used for the analysis of nuclear materials in environmental samples mainly for the safeguards, in addition to the Comprehensive Nuclear-Test-Ban Treaty verification and research on environmental sciences. The CLEAR facility was designed to meet conflicting requirements of a cleanroom and for handling of nuclear materials according to Japanese regulations, i.e., to avoid contamination from outside and to contain nuclear materials inside the facility. This facility has been intended to be used for wet chemical treatment, instrumental analysis and particle handling. A fume-hood to provide a clean work surface for handling of nuclear materials was specially designed. Much attention was paid to the selection of construction materials for use to corrosive acids. The performance of the cleanroom and analytical background in the laboratory are discussed. This facility has satisfactory specification required for joining the International Atomic Energy Agency Network of Analytical Laboratories. It can be concluded that the CLEAR facility enables analysis of ultra trace amounts of nuclear materials at sub-pictogram level in environmental samples. (author)

  19. k0-INAA of Venezuelan ceramics and complete statistical analysis to establish their provenance

    International Nuclear Information System (INIS)

    A group of 46 archaeological figurines samples (ad 1300 and 1500) from Venezuelan mainland and northern island were analyzed by k0-instrumental neutron activation analysis (k0-INAA) to obtain their elemental content and give a step ahead to establish the provenance of the island figurines. In total 37 elemental concentrations were measured with uncertainties between 3 and 20 %. To make the study of provenance, a complete statistical analysis was achieved; Fisher linear discriminant, principal component analysis, hierarchical clustering and the Hotelling T2 test were used to this end. Furthermore, not only the 46 samples analyzed in this work by k0-INAA were used, but also 40 samples analyzed by PGNAA and reported by Sajo-Bohus et al. (JRNC 265(2):247-256, 2005) were included in the statistical analysis. It was done in order to increase the size of the data set, and then to obtain from the statistical techniques more reliable results. It was found that a very good differentiation exits between the figurines from the island and from the mainland supporting the idea that the raw materials of the figurines come from different places. (author)

  20. 不同毒力结核分支杆菌感染RAW264.7巨噬细胞不同时间细胞因子转录水平标准曲线的建立及应用%Establishment and Application a Standard Curve of Quantitative Real-time PCR for Cytokine Transcript Levels after Infected by Different Virulence Mycobacterium tuberculosis at Different Time Point

    Institute of Scientific and Technical Information of China (English)

    张妍; 宋纪伟; 曾范利; 时坤; 李健明; 刘杨; 刘菲; 孙凡婷; 杜锐

    2014-01-01

    In order to detect the change of cytokine transcript levels after infected by Mycobacterium tuberculosis, build recombinant standard plasmids of IL-6, IL-10,TNF-α, a quantitative real-time PCR standard curve was established for cytokine transcript levels. Application of the method was used to analyze cytokine transcript levels that different virulence Mycobacterium tuberculosis infecting RAW264. 7 macrophages at six time points. Comparison with the control group,H37Rv and BCG made changes in transcript levels of these three cytokines, while the IL-6 and TNF-α have more obvious varies. This study provided new ideas of mechanism between Mycobacterium tuberculosis and macrophages through different time points cytokines transcriptional level.%为检测不同毒力结核分枝杆菌感染巨噬细胞后细胞因子转录水平的变化,构建IL-6、IL-10、TNF-α重组标准质粒,建立了实时荧光定量PCR标准曲线。应用该方法对不同毒力结核分枝杆菌感染RAW264.7巨噬细胞6个时间点细胞因子的转录水平进行分析,结果显示H37Rv组、BCG组相比于对照组刺激后均使三种细胞因子的转录水平发生变化,其中IL-6和TNF-α发生明显变化。本试验通过对不同时间点细胞因子转录水平的分析为结核分枝杆菌对巨噬细胞凋亡机制的研究提供新思路。

  1. Utilizing job/task analysis to establish content validity in the design of training programs

    International Nuclear Information System (INIS)

    The decade of the 1980's has been a turbulent time for the Department of Energy. With concern mounting about the terrorist threat, a wave of congressional inquiries and internal inspections crossed the nation and engulfed many of the nuclear laboratories and facilities operated by DOE contractors. A typical finding was the need to improve, and increase, the training of the protective force. The immediate reaction resulted in a wide variety of responses, with most contractors feeling safer with too much, rather than not enough training. As soon as the initial pressures to upgrade subsided, a task force was established to evaluate the overall training needs. Representatives from the contractor facilities worked together to conduct a job analysis of the protective force. A generic task inventory was established, and validated at the different sites. This list has been invaluable for determining the tasks, conditions, and standards needed to develop well stated learning objectives. The enhanced training programs are being refined to ensure job content validity based on the data collected

  2. Establishment of a clean chemistry laboratory at JAERI. Clean laboratory for environmental analysis and research (CLEAR)

    Energy Technology Data Exchange (ETDEWEB)

    Hanzawa, Yukiko; Magara, Masaaki; Watanabe, Kazuo [Japan Atomic Energy Research Inst., Tokai, Ibaraki (Japan). Tokai Research Establishment] [and others

    2003-02-01

    The JAERI has established a facility with a cleanroom: the Clean Laboratory for Environmental Analysis and Research (CLEAR). This report is an overview of the design, construction and performance evaluation of the CLEAR in the initial stage of the laboratory operation in June 2001. The CLEAR is a facility to be used for analyses of ultra trace amounts of nuclear materials in environmental samples for the safeguards, for the CTBT verification and for researches on environmental sciences. One of the special features of the CLEAR is that it meets double requirements of a cleanroom and for handling of nuclear materials. As another feature of the CLEAR, much attention was paid to the construction materials of the cleanroom for trace analysis of metal elements using considerable amounts of corrosive acids. The air conditioning and purification system, specially designed experimental equipment to provide clean work surfaces, utilities and safety systems are also demonstrated. The potential contamination from the completed cleanroom atmosphere during the analytical procedure was evaluated. It can be concluded that the CLEAR has provided a suitable condition for reliable analysis of ultra trace amounts of nuclear materials and other heavy elements in environmental samples. (author)

  3. A framework for establishing the technical efficiency of Electricity Distribution Counties (EDCs) using Data Envelopment Analysis

    International Nuclear Information System (INIS)

    Highlights: • Six models are employed to establish the technical efficiency of Electricity Distribution Counties. • A diagnostic parameter is incorporated to account for differences across Electricity Distribution Counties. • The amalgamation of Electricity Distribution Counties leads to improved efficiency in the production of energy. - Abstract: European Energy market liberalization has entailed the restructuring of electricity power markets through the unbundling of electricity generation, transmission and distribution, supply activities and introducing competition into electricity generation. Under these new electricity market regimes, it is important to have an evaluation tool that is capable of examining the impacts of these market changes. The adoption of Data Envelopment Analysis as a form of benchmarking for electricity distribution regulation is one method to conduct this analysis. This paper applies a Data Envelopment Analysis framework to the electricity distribution network in Ireland to explore the merits of using this approach, to determine the technical efficiency and the potential scope for efficiency improvements through reorganizing and the amalgamation of the distribution network in Ireland. The results presented show that overall grid efficiency is improved through this restructuring. A diagnostic parameter is defined and pursued to account for aberrations across Electricity Distribution Counties as opposed to the traditionally employed environmental variables. The adoption of this diagnostic parameter leads to a more intuitive understanding of Electricity Distribution Counties

  4. Towards a framework for establishing rigour in a discourse analysis of midwifery professionalisation.

    Science.gov (United States)

    Nixon, Anne; Power, Charmaine

    2007-03-01

    This paper develops a framework for establishing rigour for a discourse analysis of professional transition in midwifery, theorised as a 'female professional project'. Discourse analysis has gained recognition as a useful approach in nursing and midwifery research. It provides an alternative to those qualitative approaches that propose to reveal a 'reality' from the perspective of the individual experience, and that this lived experience can be directly represented in language. There are multiple discourse analytic approaches, and often researchers are not explicit about what type they are employing. Furthermore, to date there are few clear guidelines for what constitutes rigour in a discourse analytic study. Rigour in qualitative research broadly falls into two categories: the replication perspective that argues for the maintenance of criteria of validity and reliability, usually with an underpinning assumption (often unquestioned) that a 'truth' can be revealed. The second is the parallel perspective that questions the ontological and epistemological assumptions of the replication perspective and argues for the development of criteria that reflect the assumed questionable nature of truth and reality. Transferring these criteria to a discourse analytic study revealed some problems. A common challenge identified in achieving rigour in discourse analysis is the maintenance of congruity between the epistemological and ontological basis of a piece of research and the actual analysis conducted or reported. A framework is proposed to address these incongruities and includes six elements. As nurses and midwives increasingly employ discourse analysis in their research, attempts to clarify how rigour will be determined are important if the outcomes of such research are to be seriously considered for their implications for theory, policy and practice.

  5. Establishment of Grain Farmers’ Supply Response Model and Empirical Analysis under Minimum Grain Purchase Price Policy

    Institute of Scientific and Technical Information of China (English)

    2012-01-01

    Based on farmers’ supply behavior theory and price expectations theory,this paper establishes grain farmers’ supply response model of two major grain varieties (early indica rice and mixed wheat) in the major producing areas,to test whether the minimum grain purchase price policy can have price-oriented effect on grain production and supply in the major producing areas. Empirical analysis shows that the minimum purchase price published annually by the government has significant positive impact on farmers’ grain supply in the major grain producing areas. In recent years,China steadily raises the level of minimum grain purchase price,which has played an important role in effectively protecting grain farmers’ interests,mobilizing the enthusiasm of farmers’ grain production,and ensuring the market supply of key grain varieties.

  6. Cloning and Binary Analysis of the PhoP Gene in the Mycobacterium Bird Regulation System%鸟分枝杆菌二元调控系统PhoP基因的克隆与分析

    Institute of Scientific and Technical Information of China (English)

    丁峰山; 何时义; 王爱妍; 付鑫; 杨东君; 凌敏

    2015-01-01

    目的:对鸟分枝杆菌调控因子PhoP的基因序列和氨基酸序列进行生物信息学分析。方法:利用PCR技术扩增鸟分枝杆菌HIV/AIDS临床分离株PhoP基因,与pEASY-T1载体连接后测序,运用生物信息学工具对其序列进行分析。结果:PCR产物显示一条<为720bp的条带,测序结果与GenBank数据库中鸟分枝杆菌104株PhoP基因序列完全一致。该分离株PhoP的基因和氨基酸序列与结核分枝杆菌H37Rv比对,同源性分别为86%和94%。分离株PhoP基因编码的氨基酸数目为239个氨基酸,其中从136到239位氨基酸为DNA结合区。结论:成功克隆出鸟分枝杆菌HIV/AIDS临床分离株的PhoP基因。该分离株PhoP的氨基酸序列与结核分枝杆菌具有高度的同源性。%Objective: To conduct the bioinformatics analysis of the Mycobacterium bird regulation PhoP gene sequences and the amino acid sequencing. Methods:PCR technique was used to amplify Mycobacterium HIV/AIDS PhoP gene. After connect-ed with pEASY-T1 vector, the sequence was tested. Bioinformatics tool was utilized to analyze the sequences. Results: PCR products showed an article of 720bp. Sequence results in the GenBank database bird showed that 104 strains of Mycobacterium PhoP gene sequences were exactly the same with those of the PhoP genes. The PhoP gene and amino acid sequences of isolates of Mycobacterium tuberculosis H37Rv were aligned, with the homology of 86% and 94% respectively. The number of amino acids isolated from PhoP gene coding is 239. And amino acids from 136 to 239 were DNA-binding domain. Conclusion:A clin-ical isolated PhoP gene of a Mycobacterium bird HIV/AIDS was successfully cloned. The PhoP amino acid sequences of it have a high degree of homology with the Mycobacterium tuberculosis.

  7. EFFECT OF SOME MEDICINAL PLANTS ON GROWTH OF MYCOBACTERIUM TUBERCULOSIS, MULTI DRUG RESISTANT MYCOBACTERIUM TUBERCULOSIS AND MYCOBACTERIUM OTHER THAN TUBERCULOSIS

    Directory of Open Access Journals (Sweden)

    Prashant Shukla

    2013-12-01

    Full Text Available Six plants of medicinal uses were tried for their inhibitory effect on Mycobacterium tuberculosis (MTB, multi drug resistant Mycobacterium tuberculosis (MDR MTB and Mycobacterium other than tuberculosis (MOTT. MTB, MDR MTB and MOTT were cultured in 12B medium vials for Bacterc 460 TB system and incubated at 37˚C. The vials were read in Bacterc 460 TB system. Garlic, Ocimum sanctum, onion and neem showed effectiveness towards Mycobacterium tuberculosis and multi drug resistant Mycobacterium tuberculosis to some extent but ginger showed no effect at all. None of the plants studied had any inhibitory effect on Mycobacterium other than tuberculosis. Aloe vera had opposite effect on the growth and it was found to be assisting the growth of Mycobacterium tuberculosis and multi drug resistant Mycobacterium tuberculosis. The tests performed were in-vitro and the authors conlude that in-vivo the results may vary.

  8. Analysis on the Drug resistance of 322 Cases of Mycobacterium Tuberculosis in A District%某区322株结核杆菌的耐药性分析

    Institute of Scientific and Technical Information of China (English)

    韩珍; 林日文; 李静

    2013-01-01

      Objective To understand the status of drug resistance in 322 Cases of Mycobacterium tuberculosis in sputum of patients with culture-positive strains isolated in Huizhou city tuberculosis institute, and to provide evidence for clinical rational chemotherapy. Methods In our hospital, 322 Cases of Mycobacterium tuberculosis in sputum with culture-positive strains isolated as the research object, with proportion method for first-line tuberculosis drugs medication to analysis the drug resistance. Results 36 cases of 322 mycobacterium tuberculosis were resistance in four kinds of anti-TB drugs, occupied 10.84%,in which S-resistance was highest, occupied 35.8%. Conclusion The drug resistance of tuberculosis mycobacterium in Huizhou area was serious, so we should strengthen the management of the clinical treatment of tuberculosis, the development of rational chemotherapy to prevent the spread of resistant bacteria.%  目的了解惠州市结核病防治院322株痰培养阳性结核杆菌的耐药情况,为临床合理用药提供理论依据。方法以我院322株痰培养阳性的结核杆菌为研究对象,用比例法进行一线结核用药的药敏实验。结果322株结核分枝杆菌对4种抗结核药物全部耐药者36例,占10.84%;以耐链霉素为最高,占35.8%。结论惠州市结核病耐药情况仍然十分严峻,应加强结核病临床治疗的管理,制定合理化疗方案,防止耐药菌传播。

  9. Establishment of LC-MS methods for the analysis of palmitoylated surfactant proteins.

    Science.gov (United States)

    Harayama, Takeshi; Shindou, Hideo; Kita, Yoshihiro; Otsubo, Eiji; Ikeda, Kazushige; Chida, Shoichi; Weaver, Timothy E; Shimizu, Takao

    2015-07-01

    The surfactant proteins (SPs), SP-B and SP-C, are important components of pulmonary surfactant involved in the reduction of alveolar surface tension. Quantification of SP-B and SP-C in surfactant drugs is informative for their quality control and the evaluation of their biological activity. Western blot analysis enabled the quantification of SP-B, but not SP-C, in surfactant drugs. Here, we report a new procedure involving chemical treatments and LC-MS to analyze SP-C peptides. The procedure enabled qualitative analysis of SP-C from different species with discrimination of the palmitoylation status and the artificial modifications that occur during handling and/or storage. In addition, the method can be used to estimate the total amount of SP-C in pulmonary surfactant drugs. The strategy described here might serve as a prototype to establish analytical methods for peptides that are extremely hydrophobic and behave like lipids. The new method provides an easy measurement of SP-C from various biological samples, which will help the characterization of various experimental animal models and the quality control of surfactant drugs, as well as diagnostics of human samples. PMID:26022805

  10. Prevalence and occurrence rate of Mycobacterium tuberculosis Haarlem family multi-drug resistant in the worldwide population: A systematic review and meta-analysis

    OpenAIRE

    Rashid Ramazanzadeh; Daem Roshani; Pegah Shakib; Samaneh Rouhi

    2015-01-01

    Background: Transmission of Mycobacterium tuberculosis (M. tuberculosis) can occur in different ways. Furthermore, drug resistant in M. tuberculosis family is a major problem that creates obstacles in treatment and control of tuberculosis (TB) in the world. One of the most prevalent families of M. tuberculosis is Haarlem, and it is associated with drug resistant. Our objectives of this study were to determine the prevalence and occurrence rate of M. tuberculosis Haarlem family multi-drug resi...

  11. Analysis of host responses to Mycobacterium tuberculosis antigens in a multi-site study of subjects with different TB and HIV infection states in sub-Saharan Africa.

    Directory of Open Access Journals (Sweden)

    Jayne S Sutherland

    Full Text Available BACKGROUND: Tuberculosis (TB remains a global health threat with 9 million new cases and 1.4 million deaths per year. In order to develop a protective vaccine, we need to define the antigens expressed by Mycobacterium tuberculosis (Mtb, which are relevant to protective immunity in high-endemic areas. METHODS: We analysed responses to 23 Mtb antigens in a total of 1247 subjects with different HIV and TB status across 5 geographically diverse sites in Africa (South Africa, The Gambia, Ethiopia, Malawi and Uganda. We used a 7-day whole blood assay followed by IFN-γ ELISA on the supernatants. Antigens included PPD, ESAT-6 and Ag85B (dominant antigens together with novel resuscitation-promoting factors (rpf, reactivation proteins, latency (Mtb DosR regulon-encoded antigens, starvation-induced antigens and secreted antigens. RESULTS: There was variation between sites in responses to the antigens, presumably due to underlying genetic and environmental differences. When results from all sites were combined, HIV- subjects with active TB showed significantly lower responses compared to both TST(- and TST(+ contacts to latency antigens (Rv0569, Rv1733, Rv1735, Rv1737 and the rpf Rv0867; whilst responses to ESAT-6/CFP-10 fusion protein (EC, PPD, Rv2029, TB10.3, and TB10.4 were significantly higher in TST(+ contacts (LTBI compared to TB and TST(- contacts fewer differences were seen in subjects with HIV co-infection, with responses to the mitogen PHA significantly lower in subjects with active TB compared to those with LTBI and no difference with any antigen. CONCLUSIONS: Our multi-site study design for testing novel Mtb antigens revealed promising antigens for future vaccine development. The IFN-γ ELISA is a cheap and useful tool for screening potential antigenicity in subjects with different ethnic backgrounds and across a spectrum of TB and HIV infection states. Analysis of cytokines other than IFN-γ is currently on-going to determine correlates of

  12. Clonal analysis of the T-cell response to in vivo expressed Mycobacterium tuberculosis protein Rv2034, using a CD154 expression based T-cell cloning method.

    Directory of Open Access Journals (Sweden)

    Susanna Commandeur

    Full Text Available Tuberculosis (TB, caused by Mycobacterium tuberculosis (Mtb, remains a leading cause of death worldwide. A better understanding of the role of CD4+ and CD8+ T cells, which are both important to TB protection, is essential to unravel the mechanisms of protection and to identify the key antigens seen by these T cells. We have recently identified a set of in vivo expressed Mtb genes (IVE-TB which is expressed during in vivo pulmonary infection in mice, and shown that their encoded antigens are potently recognized by polyclonal T cells from tuberculin skin test-positive, in vitro ESAT-6/CFP10-responsive individuals. Here we have cloned T cells specific for one of these newly identified in vivo expressed Mtb (IVE-TB antigens, Rv2034. T cells were enriched based on the expression of CD154 (CD40L, which represents a new method for selecting antigen-specific (low frequency T cells independent of their specific function. An Rv2034-specific CD4+ T-cell clone expressed the Th1 markers T-bet, IFN-γ, TNF-α, IL-2 and the cytotoxicity related markers granzyme B and CD107a as measured by flow cytometry. The clone specifically recognized Rv2034 protein, Rv2034 peptide p81-100 and Mtb lysate. Remarkably, while the recognition of the dominant p81-100 epitope was HLA-DR restricted, the T-cell clone also recognized a neighboring epitope (p88-107 in an HLA-DR- as well as HLA-DQ1-restricted fashion. Importantly, the T-cell clone was able to inhibit Mtb outgrowth from infected monocytes significantly. The characterization of the polyfunctional and Mtb inhibitory T-cell response to IVE-TB Rv2034 at the clonal level provides detailed further insights into the potential of IVE-TB antigens as new vaccine candidate antigens in TB. Our new approach allowed the identification of T-cell subsets that likely play a significant role in controlling Mtb infection, and can be applied to the analysis of T-cell responses in patient populations.

  13. Gene expression analysis approach to establish possible links between Parkinson's disease, cancer and cardiovascular diseases.

    Science.gov (United States)

    Karim, Sajjad; Mirza, Zeenat; Kamal, Mohammad A; Abuzenadah, Adel M; Al-Qahtani, Mohammed H

    2014-01-01

    Non-communicable chronic diseases have been apparently established as threat to human health, and are currently the world's main killer. Cardiovascular diseases (CVD), cancer, diabetes and neurodegenerative diseases are collectively amounting to more than 60% of non-communicable disease burden across world. Tremendous advancements in healthcare enabled us to fight several health problems primarily infectious diseases. However, this increased longevity where in many cases an individual suffers from several such chronic diseases simultaneously, making treatment complex. Finding whether diseases can coexist in an individual by chance or there exists a possible association between them is vital. Our goal is to establish possible existing link among CVD, cancer and Parkinson's disease (PD) for better understanding of the associated molecular network. In this study, we integrated multiple dataset retrieved from the National Centre for Biotechnology Information's Gene Expression Omnibus database, and took a systems-biology approach to compare and distinguish the molecular network associated with PD, cancer and CVD. We identified 230, 308 and 1619 differentially expressed genes for CVD, cancer and PD dataset respectively using cut off p value2. We integrated these data with known pathways using Ingenuity Pathway Analysis tool and found following common pathways associated with all three diseases to be most affected; epithelial adherens junction signaling, remodelling of epithelial adherens junctions, role of BRCA1 in DNA damage response, sphingomyelin metabolism, 3- phosphoinositide biosynthesis, acute myeloid leukemia signaling, type I diabetes mellitus signaling, agrin interactions at neuromuscular junction, role of IL-17A in arthritis, and antigen presentation pathways. In conclusion, CVD, cancer and PD appear tightly associated at molecular level.

  14. Characteristic and functional analysis of a newly established porcine small intestinal epithelial cell line.

    Directory of Open Access Journals (Sweden)

    Jing Wang

    Full Text Available The mucosal surface of intestine is continuously exposed to both potential pathogens and beneficial commensal microorganisms. Recent findings suggest that intestinal epithelial cells, which once considered as a simple physical barrier, are a crucial cell lineage necessary for maintaining intestinal immune homeostasis. Therefore, establishing a stable and reliable intestinal epithelial cell line for future research on the mucosal immune system is necessary. In the present study, we established a porcine intestinal epithelial cell line (ZYM-SIEC02 by introducing the human telomerase reverse transcriptase (hTERT gene into small intestinal epithelial cells derived from a neonatal, unsuckled piglet. Morphological analysis revealed a homogeneous cobblestone-like morphology of the epithelial cell sheets. Ultrastructural indicated the presence of microvilli, tight junctions, and a glandular configuration typical of the small intestine. Furthermore, ZYM-SIEC02 cells expressed epithelial cell-specific markers including cytokeratin 18, pan-cytokeratin, sucrase-isomaltase, E-cadherin and ZO-1. Immortalized ZYM-SIEC02 cells remained diploid and were not transformed. In addition, we also examined the host cell response to Salmonella and LPS and verified the enhanced expression of mRNAs encoding IL-8 and TNF-α by infection with Salmonella enterica serovars Typhimurium (S. Typhimurium. Results showed that IL-8 protein expression were upregulated following Salmonella invasion. TLR4, TLR6 and IL-6 mRNA expression were upregulated following stimulation with LPS, ZYM-SIEC02 cells were hyporeponsive to LPS with respect to IL-8 mRNA expression and secretion. TNFα mRNA levels were significantly decreased after LPS stimulation and TNF-α secretion were not detected challenged with S. Typhimurium neither nor LPS. Taken together, these findings demonstrate that ZYM-SIEC02 cells retained the morphological and functional characteristics typical of primary swine

  15. Mutational analysis of Mycobacterium UvrD1 identifies functional groups required for ATP hydrolysis, DNA unwinding, and chemomechanical coupling

    OpenAIRE

    Sinha, Krishna Murari; Glickman, Michael S.; Shuman, Stewart

    2009-01-01

    Mycobacterial UvrD1 is a DNA-dependent ATPase and a Ku-dependent 3’ to 5’ DNA helicase. The UvrD1 motor domain resembles that of the prototypal superfamily I helicases UvrD and PcrA. Here we performed a mutational analysis of UvrD1 guided by the crystal structure of a DNA-bound E. coli UvrD-ADP-MgF3 transition state mimetic. Alanine scanning and conservative substitutions identified five amino acids essential for both ATP hydrolysis and duplex unwinding, including those implicated in phosphoh...

  16. Mycobacterium avium subspecies paratuberculosis infects and multiplies in enteric glial cells

    Institute of Scientific and Technical Information of China (English)

    2007-01-01

    AIM: To establish the role of enteric glial cells duringinfection with Mycobacterium avium subspeciesparatuberculosis (MAP) in Crohn's disease.METHODS: In order to establish the role of enteric glial cells during infection with M. avium subspecies paratuberculosis (MAP) in Crohn's disease, Map adhesion experiments on enteric glial cells were performed as well as expression analysis of Map sigma factors during infection.RESULTS: In this study, for the first time, we found a high affinity of MAP to enteric glial cells and we analyzed the expression of MAP sigma factors under different conditions of growth.CONCLUSION: The fact that Map showed a high affinity to the glial cells raises concerns about the complicated etiology of the Crohn's disease. Elucidation of the mechanisms whereby inflammation alters enteric neural control of gut functions may lead to novel treatments for Crohn's disease.

  17. Bacteriophage- based tests for the detection of Mycobacterium tuberculosis in clinical specimens: a systematic review and meta- analysis

    Directory of Open Access Journals (Sweden)

    Pascopella Lisa

    2005-07-01

    Full Text Available Abstract Background Sputum microscopy, the most important conventional test for tuberculosis, is specific in settings with high burden of tuberculosis and low prevalence of non tuberculous mycobacteria. However, the test lacks sensitivity. Although bacteriophage-based tests for tuberculosis have shown promising results, their overall accuracy has not been systematically evaluated. Methods We did a systematic review and meta-analysis of published studies to evaluate the accuracy of phage-based tests for the direct detection of M. tuberculosis in clinical specimens. To identify studies, we searched Medline, EMBASE, Web of science and BIOSIS, and contacted authors, experts and test manufacturers. Thirteen studies, all based on phage amplification method, met our inclusion criteria. Overall accuracy was evaluated using forest plots, summary receiver operating (SROC curves, and subgroup analyses. Results The data suggest that phage-based assays have high specificity (range 0.83 to 1.00, but modest and variable sensitivity (range 0.21 to 0.88. The sensitivity ranged between 0.29 and 0.87 among smear-positive, and 0.13 to 0.78 among smear-negative specimens. The specificity ranged between 0.60 and 0.88 among smear-positive and 0.89 to 0.99 among smear-negative specimens. SROC analyses suggest that overall accuracy of phage-based assays is slightly higher than smear microscopy in direct head-to-head comparisons. Conclusion Phage-based assays have high specificity but lower and variable sensitivity. Their performance characteristics are similar to sputum microscopy. Phage assays cannot replace conventional diagnostic tests such as microscopy and culture at this time. Further research is required to identify methods that can enhance the sensitivity of phage-based assays without compromising the high specificity.

  18. Mutational analysis of Mycobacterium UvrD1 identifies functional groups required for ATP hydrolysis, DNA unwinding, and chemomechanical coupling.

    Science.gov (United States)

    Sinha, Krishna Murari; Glickman, Michael S; Shuman, Stewart

    2009-05-19

    Mycobacterial UvrD1 is a DNA-dependent ATPase and a Ku-dependent 3' to 5' DNA helicase. The UvrD1 motor domain resembles that of the prototypal superfamily I helicases UvrD and PcrA. Here we performed a mutational analysis of UvrD1 guided by the crystal structure of a DNA-bound Escherichia coli UvrD-ADP-MgF(3) transition state mimetic. Alanine scanning and conservative substitutions identified amino acids essential for both ATP hydrolysis and duplex unwinding, including those implicated in phosphohydrolase chemistry via transition state stabilization (Arg308, Arg648, Gln275), divalent cation coordination (Glu236), or activation of the nucleophilic water (Glu236, Gln275). Other residues important for ATPase/helicase activity include Phe280 and Phe72, which interact with the DNA 3' single strand tail. ATP hydrolysis was uncoupled from duplex unwinding by mutations at Glu609 (in helicase motif V), which contacts the ATP ribose sugar. Introducing alanine in lieu of the adenine-binding "Q motif" glutamine (Gln24) relaxed the substrate specificity in NTP hydrolysis, e.g., eliciting a gain of function as a UTPase/TTPase, although the Q24A mutant still relied on ATP/dATP for duplex unwinding. Our studies highlight the role of the Q motif as a substrate filter and the contributions of adenosine-binding residues as couplers of NTP hydrolysis to motor activity. The Ku-binding function of UvrD1 lies within its C-terminal 270 amino acid segment. Here we found that deleting the 90 amino acid C-terminal domain, which is structurally uncharacterized, diminished DNA unwinding, without affecting ATP hydrolysis or binding to the DNA helicase substrate, apparently by affecting the strength of the UvrD1-Ku interaction. PMID:19317511

  19. Extraction of Mycobacterium tuberculosis DNA: a Question of Containment

    OpenAIRE

    Somerville, Wendy; Thibert, Louise; Schwartzman, Kevin; Behr, Marcel A.

    2005-01-01

    DNA fingerprinting of Mycobacterium tuberculosis by IS6110 restriction fragment length polymorphism analysis requires substantial high-quality DNA. We demonstrated that, despite extraction treatments that might be expected to inactivate this organism, M. tuberculosis remained viable during this process. These data suggest that the extraction of M. tuberculosis DNA should be performed within containment until complete.

  20. ANALYSIS OF THE POSSIBILITY FOR ESTABLISHING PROJECT MANAGEMENT OFFICE (PMO IN COMPANIES IN SERBIA

    Directory of Open Access Journals (Sweden)

    Dragana Milin

    2012-09-01

    Full Text Available Project Management Office (PMO is an organizational unit established to help project managers, project teams and the various levels of management in carrying out the principles of project management. The research was carried out in Serbia, in 2011, with the aim to establish which methodologies and techniques are used for project m anagement, and which of them are used the most frequently. Furthermore, the need for establishment of PMOs in Serbia is discussed. These offices should help in establishing a standardized methodology (at the organization level and thus overcome the obviou s poor use of any project management methodology at all.

  1. Analysis on drug resistance of non-tuberculosis mycobacterium in Sichuan province%非结核分枝杆菌耐药情况分析

    Institute of Scientific and Technical Information of China (English)

    李定越; 饶正远; 杨筠; 张旭东; 刘兴菊; 郑德福; 蒋绍双

    2012-01-01

    目的 探讨非结核分枝杆菌(non-tuberculosis mycobacterium,以下简称NTM)的流行病学特征,观察和分析NTM的耐药情况,为NTM肺病的防治策略提供科学依据.方法 对2006~2010年期间收集的分枝杆菌进行药物敏感性测定、菌种鉴定,对鉴定为NTM的病例进行综合分析.结果 5年间对765个菌株进行了结核分枝杆菌和非结核分枝杆菌的菌型鉴,确诊为NTM的为79株(10.32%),其中77株对异烟肼、对氨基水杨酸钠、乙胺丁醇、利福平、链霉素等常用抗结核药物存在不同程度的耐药,耐药率高达97.46%,且大多菌株同时对多种抗结核药物耐药.结论 NTM对抗结核药物具极高的耐药性.为此,对可疑肺结核患者应尽可能进行痰分离培养和多种抗结核药物敏感性测定.研究、探索抗NTM的新药及更为有效的诊断方法是当务之急.%Objective To discuss the epidemiology character of non-tuberculosis mycobacterium( hereinafter referred to as NTM) , analyze drug resistance and provide scientific evidence for the NTM control strategies. Methods The positive mycobacte-riums collected from 2006 to 2010 were detected by tests of drug susceptibility and strain identification, and we analyzed the cases with NTM. Results 765 strains of positive mycobacterium were separated into tuberculosis mycobacterium composite group and non-tuberculosis mycobacterium, 79 strains were identified as NTM, 10. 32% of total strains;77 strains had drug resistance with varying degrees to isoniazid, paminosalicylic acid, ethambutol, rifampicin, streptomycin. The rate of drug resistance was 97.46% , and most of the strains had drug resistance to various antituberculosis drugs. Conclusion NTM have strong drug resistance to antituberculosis drugs. So we need to conduct isolation and culture of the sputum of the suspicious patients with Tuberculosis, and detect drug susceptibility of the various antituberculosis drugs, to research new drugs against

  2. Analysis of newly established EST databases reveals similarities between heart regeneration in newt and fish

    Directory of Open Access Journals (Sweden)

    Weis Patrick

    2010-01-01

    Full Text Available Abstract Background The newt Notophthalmus viridescens possesses the remarkable ability to respond to cardiac damage by formation of new myocardial tissue. Surprisingly little is known about changes in gene activities that occur during the course of regeneration. To begin to decipher the molecular processes, that underlie restoration of functional cardiac tissue, we generated an EST database from regenerating newt hearts and compared the transcriptional profile of selected candidates with genes deregulated during zebrafish heart regeneration. Results A cDNA library of 100,000 cDNA clones was generated from newt hearts 14 days after ventricular injury. Sequencing of 11520 cDNA clones resulted in 2894 assembled contigs. BLAST searches revealed 1695 sequences with potential homology to sequences from the NCBI database. BLAST searches to TrEMBL and Swiss-Prot databases assigned 1116 proteins to Gene Ontology terms. We also identified a relatively large set of 174 ORFs, which are likely to be unique for urodele amphibians. Expression analysis of newt-zebrafish homologues confirmed the deregulation of selected genes during heart regeneration. Sequences, BLAST results and GO annotations were visualized in a relational web based database followed by grouping of identified proteins into clusters of GO Terms. Comparison of data from regenerating zebrafish hearts identified biological processes, which were uniformly overrepresented during cardiac regeneration in newt and zebrafish. Conclusion We concluded that heart regeneration in newts and zebrafish led to the activation of similar sets of genes, which suggests that heart regeneration in both species might follow similar principles. The design of the newly established newt EST database allows identification of molecular pathways important for heart regeneration.

  3. Scaling Analysis Techniques to Establish Experimental Infrastructure for Component, Subsystem, and Integrated System Testing

    Energy Technology Data Exchange (ETDEWEB)

    Sabharwall, Piyush [Idaho National Laboratory (INL), Idaho Falls, ID (United States); O' Brien, James E. [Idaho National Laboratory (INL), Idaho Falls, ID (United States); McKellar, Michael G. [Idaho National Laboratory (INL), Idaho Falls, ID (United States); Housley, Gregory K. [Idaho National Laboratory (INL), Idaho Falls, ID (United States); Bragg-Sitton, Shannon M. [Idaho National Laboratory (INL), Idaho Falls, ID (United States)

    2015-03-01

    Hybrid energy system research has the potential to expand the application for nuclear reactor technology beyond electricity. The purpose of this research is to reduce both technical and economic risks associated with energy systems of the future. Nuclear hybrid energy systems (NHES) mitigate the variability of renewable energy sources, provide opportunities to produce revenue from different product streams, and avoid capital inefficiencies by matching electrical output to demand by using excess generation capacity for other purposes when it is available. An essential step in the commercialization and deployment of this advanced technology is scaled testing to demonstrate integrated dynamic performance of advanced systems and components when risks cannot be mitigated adequately by analysis or simulation. Further testing in a prototypical environment is needed for validation and higher confidence. This research supports the development of advanced nuclear reactor technology and NHES, and their adaptation to commercial industrial applications that will potentially advance U.S. energy security, economy, and reliability and further reduce carbon emissions. Experimental infrastructure development for testing and feasibility studies of coupled systems can similarly support other projects having similar developmental needs and can generate data required for validation of models in thermal energy storage and transport, energy, and conversion process development. Experiments performed in the Systems Integration Laboratory will acquire performance data, identify scalability issues, and quantify technology gaps and needs for various hybrid or other energy systems. This report discusses detailed scaling (component and integrated system) and heat transfer figures of merit that will establish the experimental infrastructure for component, subsystem, and integrated system testing to advance the technology readiness of components and systems to the level required for commercial

  4. Establishment of a Multiplex Real-Time Fluorescence Quantitative PCR Assay for Detection of Brucella and Mycobacterium tuberculosis%梅迪-维斯纳病毒和羊痘病毒多联实时定量PCR检测方法的建立及初步应用

    Institute of Scientific and Technical Information of China (English)

    徐军; 孙志华; 刘娟; 孟茹; 戴莉; 段晓东; 叶志辉

    2012-01-01

    To establish a method of multiplex real-time fluorescence quantitative PCR assay for fast diagnosis of Maedi-Visna virus (MVV) and Capripox virus (CPV). We designed and synthesized primers of MVV and CPV genes according to gene sequence published in GenBank,established multiplex RTFQ-PCR,then tested its stability,specificity and sensitivity,and detected the clinic and imitation samples with this method. The Tm of multiplex RT-PCR to amplify brucella and mycobacterium tuberculosis was 89~90 ℃ and 91~92 'C. But the results of the amplification of other bacteria were negative. The lowest detection limit for DNA of Brucella,MVV and CPV was 25 copies/μL,40 copies/μL,80 copies/μL,respectively. In conclusion,the assay could be used to detect CPV simultaneously.%利用多联实时荧光定量PCR技术建立了一种梅迪-维斯纳病毒和羊痘病毒快速鉴别诊断方法.分别设计并合成梅迪-维斯纳病毒和羊痘病毒基因的引物,建立多联实时定量PCR快速鉴别诊断方法;对所建立的方法进行稳定性、特异性和敏感性试验;并用所建立的方法对临床样品进行检测.结果显示:设计的引物敏感性和特异性较好,该多联实时荧光定量PCR方法中梅迪-维斯纳病毒Tm值为89~90℃,羊痘病毒Tm值为91~92℃,对其他供试的菌株则为阴性,并且该方法对梅迪-维斯纳病毒的DNA最低检出量为25拷贝/μL,羊痘病毒为40拷贝/μL,两病原都存在时为80拷贝/μL.研究结果表明本实验建立的方法可用于同时检测梅迪-维斯纳病毒和羊痘病毒,为动物检疫提供了一种有效的检测方法.

  5. Feasibility Analysis of Establishing Multilateral Nuclear Approaches (MNAs in the Asian Region and the Middle East

    Directory of Open Access Journals (Sweden)

    Makiko Tazaki

    2014-12-01

    Full Text Available To establish frameworks for multilateral nuclear approaches (MNAs, we identified challenges and their possible solutions through case studies proposing to establish three different MNAs, comprising existing states in the Asian region and the Middle East, in accordance with twelve features deemed necessary for establishing MNAs. In all case studies, political instability of MNA member states and the region, as well as political conflicts between MNA member states and other states were seen as challenges hindering the establishment of MNAs. There are no simple measures to overcome such challenges, but additional case-by-case measures, including the direct involvement of international organizations, supplier states and nuclear weapon states, in MNAs, as well as the application of regional safeguards and regional systems of accounting for and control of nuclear material (RSAC within MNAs, may contribute toward mitigating the political challenges.

  6. Molecular characteristics of "Mycobacterium canettii" the smooth Mycobacterium tuberculosis bacilli.

    NARCIS (Netherlands)

    Fabre, M.; Hauck, Y.; Soler, C.; Koeck, J.L.; Ingen, J. van; Soolingen, D. van; Vergnaud, G.; Pourcel, C.

    2010-01-01

    Since the first discovery of the smooth tubercle (SmTB) bacilli "Mycobacterium canettii" less than 60 isolates have been reported, all but one originating from a limited geographical location, the Horn of Africa. In spite of its rarity, the SmTB lineage deserves special attention. Previous investiga

  7. Cladistic analysis of Hemirhipini with establishment of Propalaus gen. nov. (Coleoptera, Elateridae, agrypninae

    Directory of Open Access Journals (Sweden)

    Sônia A. Casari

    2008-01-01

    Full Text Available This article includes a cladistic analysis of the tribe Hemirhipini. Are included 20 Hemirhipini genera (sensu Casari-Chen 1994, Saltamartinus Casari (1996b (Hemirhipini, 6 genera excluded from Hemirhipini and kept in Agrypninae (formerly Pyrophorinae (Casari-Chen 1993 and also, Aphileus Candèze (1857, Pyrophorus Billberg (1820 and Thoramus Sharp (1877. The type-species of the majority of genera and all species of the American genera (except Saltamartinus viduus (Chevrolat 1867 are included. This analysis demonstrates that 30 genera belong to Hemirhipini: Abiphis Fleutiaux (1926, Alaolacon Candèze (1865, Alaomorphus Hauser (1900, Alaus Eschscholtz (1829, Aliteus Candèze (1857, Anthracalaus Fairmaire (1888, Aphileus Candèze (1857, Austrocalais Neboiss (1967, Calais Castelnau (1836, Catelanus Fleutiaux (1942, Chalcolepidius Eschscholtz (1829, Chalcolepis Candèze (1857, Conobajulus Van Zwaluwenburg (1940, Coryleus Fleutiaux (1942, Cryptalaus Ôhira (1967, Eleuphemus Hyslop (1921, Eumoeus Candèze (1874, Fusimorphus Fleutiaux (1942, Hemirhipus Latreille (1829, Lacais Fleutiaux (1942, Lycoreus Candèze (1857, Mocquerysia Fleutiaux (1899, Neocalais Girard (1971, Pherhimius Fleutiaux (1942, Phibisa Fleutiaux (1942, Propalaus gen. nov., Pseudocalais Girard (1971, Saltamartinus Casari (1996, Tetrigus Candèze (1857 and Thoramus Sharp (1877. The species included in Alaus do not make a monophyletic group and Propalaus gen. nov. is established to include Alaus alicii (Pjatakowa 1941 and A. haroldi (Candèze 1878. A description of Propalaus gen. nov. (type-species: Chalcolepidius haroldi Candèze, 1878 and a new key to Hemirhipini genera are also presented.Esse artigo apresenta uma análise cladística da tribo Hemirhipini. Estão incluídos na análise, 20 gêneros de Hemirhipini (sensu Casari-Chen 1994, Saltamartinus Casari (1996b (Hemirhipini, 6 gêneros excluídos de Hemirhipini e mantidos em Pyrophorinae (= Agrypninae (Casari-Chen 1993 e tamb

  8. Identification of Secretory Proteins in Mycobacterium tuberculosis Using Pseudo Amino Acid Composition.

    Science.gov (United States)

    Yang, Huan; Tang, Hua; Chen, Xin-Xin; Zhang, Chang-Jian; Zhu, Pan-Pan; Ding, Hui; Chen, Wei; Lin, Hao

    2016-01-01

    Tuberculosis is killing millions of lives every year and on the blacklist of the most appalling public health problems. Recent findings suggest that secretory protein of Mycobacterium tuberculosis may serve the purpose of developing specific vaccines and drugs due to their antigenicity. Responding to global infectious disease, we focused on the identification of secretory proteins in Mycobacterium tuberculosis. A novel method called MycoSec was designed by incorporating g-gap dipeptide compositions into pseudo amino acid composition. Analysis of variance-based technique was applied in the process of feature selection and a total of 374 optimal features were obtained and used for constructing the final predicting model. In the jackknife test, MycoSec yielded a good performance with the area under the receiver operating characteristic curve of 0.93, demonstrating that the proposed system is powerful and robust. For user's convenience, the web server MycoSec was established and an obliging manual on how to use it was provided for getting around any trouble unnecessary. PMID:27597968

  9. Diterpene production in Mycobacterium tuberculosis

    Science.gov (United States)

    Prach, Lisa; Kirby, James; Keasling, Jay D.; Alber, Tom

    2011-01-01

    Diterpenes are a structurally diverse class of molecules common in plants, although they are very rarely found in bacteria. We report the identification in Mycobacterium tuberculosis (Mtb) of three diterpenes proposed to promote phagolysosome maturation arrest. MS analysis reveals that these diterpenes are novel compounds not previously identified in other organisms. The diterpene with highest abundance in Mtb has a mass fragmentation pattern identical to edaxadiene, which is produced in vitro from geranylgeranyl diphosphate by the enzymes Rv3377c and Rv3378c [Mann FM et al. (2009) J Am Chem Soc 131, 17526–17527]. A second diterpene found in Mtb has a similar mass spectrum, and is always observed in the same proportion relative to edaxadiene, indicating that it is a side product of the Rv3378c reaction in vivo. We name this second diterpene olefin edaxadiene B. The least abundant of the three diterpenes in Mtb extracts is tuberculosinol, a dephosphorylated side-product of the edaxadiene pathway intermediate produced by Rv3377c [Nakano C et al. (2009) Chembiochem 10, 2060–2071; Nakano C et al. (2005) Chem Commun (Camb) 8, 1016–1018]. A frameshift in Rv3377c in Mtb completely eliminates diterpene production, whereas expression of Rv3377c and Rv3378c in the nonpathogenic M. smegmatis is sufficient to produce edaxadiene and edaxadiene B. These studies define the pathway of edaxadiene and edaxadiene B biosynthesis in vivo. Rv3377c and Rv3378c are unique to Mtb and M. bovis, making them candidates for selective therapeutics and diagnostics. PMID:20670276

  10. Genotypic characteristics of a Mycobacterium sp. isolated from yellowtail Seriola quinqueradiata and striped jack Pseudocaranx dentex in Japan.

    Science.gov (United States)

    Imajoh, Masayuki; Sugiura, Hidehiro; Hashida, Yumiko; Hatai, Kishio; Oshima, Syun-ichirou; Daibata, Masanori; Kawai, Kenji

    2013-01-01

    In Japan, a Mycobacterium marinum-like mycobacterium was isolated from the yellowtail, Seriola quinqueradiata. The species was identified as M. marinum by a commercial mycobacterial DNA-DNA hybridization kit. Nevertheless, PCR restriction analysis of the DNA of its RNA polymerase β-subunit gene definitively showed that this Mycobacterium sp. was M. ulcerans. PCR analysis revealed the genotypic characteristics of M. ulcerans in the Mycobacterium sp., only the mup053 gene sequence being absent, as has been found previously in other piscine mycobacteria such as M. marinum strains DL240490 and DL045 and M. pseudoshottsii. With one exception, this Mycobacterium sp. and M. pseudoshottsii had identical 16S rRNA gene sequences, which is also probably true of M. marinum strains DL240490 and DL045. Similarly, according to comparisons of the 16S rRNA gene, ITS region, and hsp65 gene sequences, this Mycobacterium sp. is more closely related to M. pseudoshottsii than to M. ulcerans or M. marinum. A PCR product of approximately 2000 bp was amplified from region of difference 9 in the Mycobacterium sp. The nucleotide sequence revealed insertion of IS2404, the sequence of which is 1366 bp long. The novel single nucleotide polymorphisms identified in this region distinguished this Mycobacterium sp. from M. marinum strain DL240490 and M. pseudoshottsii. The present findings raise the possibility that these species have a common ancestor. Further studies are required to improve our understanding of the relationship between their geographical origin and genetic diversity. PMID:23043488

  11. Analysis of ISO/IEC 17025 for establishment of KOLAS (Korea Laboratory Accreditation Scheme) quality assurance system

    International Nuclear Information System (INIS)

    Besides one existent accredited lab, radioactive material chemical analysis lab, five test laboratories and two calibration labs are under plan to acquire the accreditation from KOLAS. But the current Quality Manual was developed according to ISO Guide 25 that was superceded by ISO/IEC 17025. Since it is tailored to the radioactive material chemical analysis lab, a number of requirements of the Manual are not applicable to the labs other than radioactive material chemical analysis lab. Through the analysis of ISO/IEC 17025, a model of quality system was established which is not only consistent with ISO/IEC 17025 but reflective of the KAERI's situation

  12. Analysis of ISO/IEC 17025 for establishment of KOLAS (Korea Laboratory Accreditation Scheme) quality assurance system

    Energy Technology Data Exchange (ETDEWEB)

    Nam, Ji Hee

    2000-12-01

    Besides one existent accredited lab, radioactive material chemical analysis lab, five test laboratories and two calibration labs are under plan to acquire the accreditation from KOLAS. But the current Quality Manual was developed according to ISO Guide 25 that was superceded by ISO/IEC 17025. Since it is tailored to the radioactive material chemical analysis lab, a number of requirements of the Manual are not applicable to the labs other than radioactive material chemical analysis lab. Through the analysis of ISO/IEC 17025, a model of quality system was established which is not only consistent with ISO/IEC 17025 but reflective of the KAERI's situation.

  13. Measuring Organizational Learning Capability in Indian Managers and Establishing Firm Performance Linkage: An Empirical Analysis

    Science.gov (United States)

    Bhatnagar, Jyotsna

    2006-01-01

    Purpose: The purpose of this research is to measure Organizational Learning Capability (OLC) perception in the managers of public, private and multinational organizations and establish the link between OLC and firm performance. Design/methodology/approach: The data were collected from a sample of 612 managers randomly drawn from Indian industry,…

  14. Genomic and transcriptomic analysis of Laccaria bicolor CAZome reveals insights into polysaccharides remodelling during symbiosis establishment

    NARCIS (Netherlands)

    Veneault-Fourrey, Claire; Commun, Carine; Kohler, Annegret; Morin, Emmanuelle; Balestrini, Raffaella; Plett, Jonathan; Danchin, Etienne; Coutinho, Pedro; Wiebenga, A.; de Vries, Ronald P; Henrissat, Bernard; Martin, Francis; van den Brink, J.

    2014-01-01

    Ectomycorrhizal fungi, living in soil forests, are required microorganisms to sustain tree growth and productivity. The establishment of mutualistic interaction with roots to form ectomycorrhiza (ECM) is not well known at the molecular level. In particular, how fungal and plant cell walls are rearra

  15. An orphan gyrB in the Mycobacterium smegmatis genome uncovered by comparative genomics

    Indian Academy of Sciences (India)

    P. Jain; V. Nagaraja

    2002-11-01

    DNA gyrase is an essential topoisomerase found in all bacteria. It is encoded by gyrB and gyrA genes. These genes are organized differently in different bacteria. Direct comparison of Mycobacterium tuberculosis and Mycobacterium smegmatis genomes reveals presence of an additional gyrB in M. smegmatis flanked by novel genes. Analysis of the amino acid sequence of GyrB from different organisms suggests that the orphan GyrB in M. smegmatis may have an important cellular role.

  16. The establishment and application of high throughput screening model targeting Mycobacterium tuberculosis peptide deformylase%以肽脱甲酰基酶为靶点的抗结核药物高通量筛选模型的建立和应用

    Institute of Scientific and Technical Information of China (English)

    张丽蓉; 赵莉莉; 魏玉珍; 李秋萍; 王莉宁; 余利岩

    2012-01-01

    目的 建立以肽脱甲酰基酶(PDF)为靶点的抗结核药物高通量筛选模型,应用该模型筛选得到活性微生物发酵液粗提物样品.方法 以结核分枝杆菌 H37Rv 基因组为模板,扩增肽脱甲酰基酶的基因片段 def,构建表达载体 pET-28a-def,表达并纯化结核分枝杆菌 PDF 酶;基于 PDF 水解三肽底物for-Met-Ala-Ser 释放出游离 NH2,而游离 NH2 可与荧光胺反应产生荧光的原理,利用测定所产生荧光值的方法,建立高通量药物筛选模型;使用该模型对 12 400 个微生物发酵液粗提物样品进行筛选,同时以耻垢分枝杆菌为检定菌,平板纸片法检测样品的抗菌活性,并检测所得阳性样品的细胞毒性.结果 成功构建了表达载体 pET-28a-def;所建立的模型稳定可行,可用于以肽脱甲酰基酶为靶点的抗结核药物的高通量筛选;用该模型对 12 400 个微生物发酵液粗提物样品进行筛选,最终得到 8 个对肽脱甲酰基酶抑制活性和抗耻垢分枝杆菌活性均较好的阳性样品,阳性率 0.06%;其中 5 个样品的细胞毒性较小.结论 建立了灵敏度好、稳定性高的结核分枝杆菌肽脱甲酰基酶抑制剂高通量药物筛选模型,应用该模型所得到的阳性样品具有进一步深入研究的意义.%Objective To establish and validate a high throughput model for screening of Mycobacterium tuberculosis peptide deformylase (PDF) inhibitors as potential antituberculosis drugs, and to perform preliminary screening with microbial fermentation extracts.Methods The H37Rv PDF coding gene def was amplified with PCR and cloned into the expression vector pET-28a. The recombinant PDF was over-expressed and purified. A high throughput model was established based on the detection of fluorescence intensity, which is caused by the reaction of fluorescamine and free NH2 released by PDF from substrate for-Met-Ala-Ser. Using the assay, 12 400 microbial fermentation extracts were screened, and

  17. The regulation of sulfur metabolism in Mycobacterium tuberculosis.

    Directory of Open Access Journals (Sweden)

    Stavroula K Hatzios

    2011-07-01

    Full Text Available Mycobacterium tuberculosis (Mtb has evolved into a highly successful human pathogen. It deftly subverts the bactericidal mechanisms of alveolar macrophages, ultimately inducing granuloma formation and establishing long-term residence in the host. These hallmarks of Mtb infection are facilitated by the metabolic adaptation of the pathogen to its surrounding environment and the biosynthesis of molecules that mediate its interactions with host immune cells. The sulfate assimilation pathway of Mtb produces a number of sulfur-containing metabolites with important contributions to pathogenesis and survival. This pathway is regulated by diverse environmental cues and regulatory proteins that mediate sulfur transactions in the cell. Here, we discuss the transcriptional and biochemical mechanisms of sulfur metabolism regulation in Mtb and potential small molecule regulators of the sulfate assimilation pathway that are collectively poised to aid this intracellular pathogen in its expert manipulation of the host. From this global analysis, we have identified a subset of sulfur-metabolizing enzymes that are sensitive to multiple regulatory cues and may be strong candidates for therapeutic intervention.

  18. Collaborative Performance Analysis and the Establishement of Main Body of Agricutlural Technology Innovation

    Institute of Scientific and Technical Information of China (English)

    2011-01-01

    Relying on the relevant data from China Statistical Yearbook and Chinese Rural Statistical Yearbook,the coolabarative performance of main body of agricutlural technology innovation is evaluated by taking quaternary-parties innovation body as the sujects and by establishing the index system from the perspectives of input and output.The results show serious dislocation of main body and bad collabrative performance.On the basis,the quaternary-parties motivation system cocerning agricutlural enterpises,agricutlural science and technology acanamy,rural hosueholds and governments is established.The system is capable of integrating market,sciencetific reserch and diffusion collection into a whole part and solving the seperation of agricutlural innovation supply and demand.

  19. Establishment of Orthotopic Xuanwei Lung Cancer SCID Mouse Model 
and Analysis of Biological Properties

    OpenAIRE

    Yongchun ZHOU; Chen, Yan; Xicai WANG; Liu, Xin; Hutao SHI; Yao, Qian; Jin, Congguo; Wu, Zhiping; Huang, Yunchao

    2012-01-01

    Background and objective The incidence of Xuanwei lung cancer ranks first in China, and its pathogenesis requires in-depth investigation. This study aims to establish an orthotopic Xuanwei lung cancer severe combined immunodeficiency (SCID) mouse model and to provide a basic experimental platform for further study. Methods The Xuanwei lung cancer cell line XWLC-05 was inoculated into the lung tissue of SCID mice in high and low doses. The tumor formation rates, tumor characteristics, spontane...

  20. Security Analysis and Improvements of Session Key Establishment for Clustered Sensor Networks

    Directory of Open Access Journals (Sweden)

    Jiye Kim

    2016-01-01

    Full Text Available WSN (wireless sensor network is one of the main technologies in IoT (Internet of Things applications or services. To date, several schemes have been proposed to establish a pair-wise key between two nodes in WSN, and most of them are designed to establish long-term keys used throughout the network lifetime. However, in the near future, if WSN will be used for information infrastructures in various fields such as manufacturing, distribution, or public facilities management and its life cycle can be as long as that of other common networks, it will definitely be advantageous in terms of security to encrypt messages using session keys instead of long-term keys. In this paper, we propose a session key establishment scheme for clustered sensor networks that is based on elliptic curve Diffie-Hellman (ECDH key exchange and hash chain. The proposed scheme eliminates vulnerabilities of existing schemes for WSN and has improved security. The proposed scheme is efficient in terms of energy costs compared to related schemes.

  1. Polymorphisms of twenty regulatory proteins between Mycobacterium tuberculosis and Mycobacterium bovis

    Science.gov (United States)

    Mycobacterium tuberculosis and Mycobacterium bovis are responsible for tuberculosis in humans or animals, respectively. Both species are closely related and belong to the Mycobacterium tuberculosis complex (MTC). M. tuberculosis is the most ancient species from which M. bovis and the other members o...

  2. Mycobacterium avium Possesses Extracellular DNA that Contributes to Biofilm Formation, Structural Integrity, and Tolerance to Antibiotics.

    Directory of Open Access Journals (Sweden)

    Sasha J Rose

    Full Text Available Mycobacterium avium subsp. hominissuis is an opportunistic pathogen that is associated with biofilm-related infections of the respiratory tract and is difficult to treat. In recent years, extracellular DNA (eDNA has been found to be a major component of bacterial biofilms, including many pathogens involved in biofilm-associated infections. To date, eDNA has not been described as a component of mycobacterial biofilms. In this study, we identified and characterized eDNA in a high biofilm-producing strain of Mycobacterium avium subsp. hominissuis (MAH. In addition, we surveyed for presence of eDNA in various MAH strains and other nontuberculous mycobacteria. Biofilms of MAH A5 (high biofilm-producing strain and MAH 104 (reference strain were established at 22°C and 37°C on abiotic surfaces. Acellular biofilm matrix and supernatant from MAH A5 7 day-old biofilms both possess abundant eDNA, however very little eDNA was found in MAH 104 biofilms. A survey of MAH clinical isolates and other clinically relevant nontuberculous mycobacterial species revealed many species and strains that also produce eDNA. RAPD analysis demonstrated that eDNA resembles genomic DNA. Treatment with DNase I reduced the biomass of MAH A5 biofilms when added upon biofilm formation or to an already established biofilm both on abiotic surfaces and on top of human pharyngeal epithelial cells. Furthermore, co-treatment of an established biofilm with DNase 1 and either moxifloxacin or clarithromycin significantly increased the susceptibility of the bacteria within the biofilm to these clinically used antimicrobials. Collectively, our results describe an additional matrix component of mycobacterial biofilms and a potential new target to help treat biofilm-associated nontuberculous mycobacterial infections.

  3. Conditional silencing of topoisomerase I gene of Mycobacterium tuberculosis validates its essentiality for cell survival.

    Science.gov (United States)

    Ahmed, Wareed; Menon, Shruti; Godbole, Adwait Anand; Karthik, Pullela V D N B; Nagaraja, Valakunja

    2014-04-01

    Topoisomerases are an important class of enzymes for regulating the DNA transaction processes. Mycobacterium tuberculosis (Mtb) is one of the most formidable pathogens also posing serious challenges for therapeutic interventions. The organism contains only one type IA topoisomerase (Rv3646c), offering an opportunity to test its potential as a candidate drug target. To validate the essentiality of M. tuberculosis topoisomerase I (TopoI(Mt) ) for bacterial growth and survival, we have generated a conditionally regulated strain of topoI in Mtb. The conditional knockdown mutant exhibited delayed growth on agar plate. In liquid culture, the growth was drastically impaired when TopoI expression was suppressed. Additionally, novobiocin and isoniazid showed enhanced inhibitory potential against the conditional mutant. Analysis of the nucleoid revealed its altered architecture upon TopoI depletion. These studies establish the essentiality of TopoI for the M. tuberculosis growth and open up new avenues for targeting the enzyme.

  4. A random sequential mechanism of aminoglycoside acetylation by Mycobacterium tuberculosis Eis protein.

    Directory of Open Access Journals (Sweden)

    Oleg V Tsodikov

    Full Text Available An important cause of bacterial resistance to aminoglycoside antibiotics is the enzymatic acetylation of their amino groups by acetyltransferases, which abolishes their binding to and inhibition of the bacterial ribosome. Enhanced intracellular survival (Eis protein from Mycobacterium tuberculosis (Mt is one of such acetyltransferases, whose upregulation was recently established as a cause of resistance to aminoglycosides in clinical cases of drug-resistant tuberculosis. The mechanism of aminoglycoside acetylation by MtEis is not completely understood. A systematic analysis of steady-state kinetics of acetylation of kanamycin A and neomycin B by Eis as a function of concentrations of these aminoglycosides and the acetyl donor, acetyl coenzyme A, reveals that MtEis employs a random-sequential bisubstrate mechanism of acetylation and yields the values of the kinetic parameters of this mechanism. The implications of these mechanistic properties for the design of inhibitors of Eis and other aminoglycoside acetyltransferases are discussed.

  5. Granulomatous hypophysitis by Mycobacterium gordonae in a non HIV-infected patient

    Directory of Open Access Journals (Sweden)

    José Luis Ruiz-Sandoval

    2009-12-01

    Full Text Available Lymphocytic or granulomatous hypophysitis is a rare entity with a difficult diagnosis. Our objective was to report a patient with non-tuberculous granulomatous hypophysitis. An HIV-negative 45-year old man with confusional state, subacute ophthalmoplegia, and clinical and laboratory findings of panhypopituitarism was seen in the emergency unit. A cranial MRI showed a sellar mass suggestive of hypophysitis. After an unsuccessful attempt with steroids and antituberculous drugs the patient died. Post-mortem histopathology revealed granulomatous lesions and restriction fragment length polymorphism analysis confirmed the presence of Mycobacterium gordonae’s DNA. In conclusion, we should consider granulomatous hypophysitis in the differential diagnosis of non-secreting hypophyseal tumors. The etiology of a pituitary granuloma by a non-tuberculous mycobacteria is best reached by histopathological techniques and molecular assays. The optimal therapy is yet to be established.

  6. Establishment, drought tolerance and recovery, and canopy analysis of turfgrasses in the transition zone

    Science.gov (United States)

    Goldsby, Anthony Lee

    Increasing water scarcity may result in greater irrigation restrictions for turfgrass. Drought tolerance and recovery of Kentucky bluegrasses ( Poa. pratensis L.) (KBG) were evaluated during and after 88 and 60 day dry downs in 2010 and 2011, respectively, under a rainout shelter. Changes in green coverage were evaluated with digital images. Green coverage declined slowest during dry downs and increased fastest during recoveries in the cultivar 'Apollo', indicating it had superior drought tolerance. Electrolyte leakage, photosynthesis, and leaf water potential were evaluated in 7 KBG cultivars during and after the dry downs. Soil moisture at 5 and 20 cm was measured. There were generally no differences in physiological parameters among cultivars during or after dry down. The highest reduction in soil moisture at 5 and 20 cm was in Apollo, suggesting it had a better developed root system for mining water from the profile during drought. Weed prevention and turfgrass establishment of 'Legacy' buffalograss (Buchloe dactyloides [Nutt.] Engelm.) and 'Chisholm' zoysiagrass (Zoysia japonica Steud.) grown on turf reinforcement mats (TRM) was evaluated. 'Chisholm' zoysiagrass stolons grew under the TRM; as such, use of TRM for this cultivar is not practical. Buffalograss had 90% or greater coverage when established on TRM in 2010 and 65% or greater coverage in 2011; coverage was similar to that in oxadiazon-treated plots at the end of each year. 'Legacy' buffalograss plugs were established on TRM over plastic for 3 weeks, stored in TRM under tree shade for 7, 14, or 21 days, and evaluated for establishment after storage. In 2010, plugs on mats stored for 7 days had similar coverage to the control, but in 2011 displayed similar coverage to plugs stored on TRM for 14 or 21 day treatments. Green leaf are index (LAI) is an important indicator of turfgrass performance, but its measurement is time consuming and destructive. Measurements using hyperspectral radiometry were

  7. Analiza dejavnikov ustanavljanja podjetja = Analysis of Factors that Promote the Establishment of the Firm

    Directory of Open Access Journals (Sweden)

    Irena Kušče

    2011-06-01

    Full Text Available After the introductory presentation of basic concepts related to entrepreneurshipand its importance, and the entrepreneur in the entrepreneurialprocess, we will focus on analyzing the factors of businessstart-up. We found that in the domestic and foreign literature there are many classifications of factors that promote the establishmentof the firm, and many different models exist as well. Most of therecent literature advocates the influence of personal factors and environmentalfactors, but in practice most of the research is based solelyon personal factors of establishment. We decided to analyze both, aswe think that this is the only possible way to provide a comprehensivetreatment of the issue.

  8. Combating highly resistant emerging pathogen Mycobacterium abscessus and Mycobacterium tuberculosis with novel salicylanilide esters and carbamates.

    Science.gov (United States)

    Baranyai, Zsuzsa; Krátký, Martin; Vinšová, Jarmila; Szabó, Nóra; Senoner, Zsuzsanna; Horváti, Kata; Stolaříková, Jiřina; Dávid, Sándor; Bősze, Szilvia

    2015-08-28

    In the Mycobacterium genus over one hundred species are already described and new ones are periodically reported. Species that form colonies in a week are classified as rapid growers, those requiring longer periods (up to three months) are the mostly pathogenic slow growers. More recently, new emerging species have been identified to lengthen the list, all rapid growers. Of these, Mycobacterium abscessus is also an intracellular pathogen and it is the most chemotherapy-resistant rapid-growing mycobacterium. In addition, the cases of multidrug-resistant Mycobacterium tuberculosis infection are also increasing. Therefore there is an urgent need to find new active molecules against these threatening strains. Based on previous results, a series of salicylanilides, salicylanilide 5-chloropyrazinoates and carbamates was designed, synthesized and characterised. The compounds were evaluated for their in vitro activity on M. abscessus, susceptible M. tuberculosis H37Rv, multidrug-resistant (MDR) M. tuberculosis MDR A8, M. tuberculosis MDR 9449/2006 and on the extremely-resistant Praha 131 (XDR) strains. All derivatives exhibited a significant activity with minimum inhibitory concentrations (MICs) in the low micromolar range. Eight salicylanilide carbamates and two salicylanilide esters exhibited an excellent in vitro activity on M. abscessus with MICs from 0.2 to 2.1 μM, thus being more effective than ciprofloxacin and gentamicin. This finding is potentially promising, particularly, as M. abscessus is a threateningly chemotherapy-resistant species. M. tuberculosis H37Rv was inhibited with MICs from 0.2 μM, and eleven compounds have lower MICs than isoniazid. Salicylanilide esters and carbamates were found that they were effective also on MDR and XDR M. tuberculosis strains with MICs ≥1.0 μM. The in vitro cytotoxicity (IC50) was also determined on human MonoMac-6 cells, and selectivity index (SI) of the compounds was established. In general, salicylanilide

  9. In vitro propagation and biochemical analysis of field established wood apple (Aegle marmelos L.

    Directory of Open Access Journals (Sweden)

    Kuldeep YADAV

    2011-05-01

    Full Text Available This paper describes an improved and rapid protocol for multiple shoot regeneration from nodal segments of wood apple (Aegle marmelos L., a medicinal tree, cultured on Murashige and Skoog (1962 (MS medium supplemented with various concentrations of auxins and cytokinins individually and in various combinations. BAP was found to be more effective than kinetin for shoot multiplication. Nodal explants responded most favorably at low BAP (2.0 mg/l producing maximum number of shoots (8.0 and uniform shoots facilitating their simultaneous harvest for rooting. The medium supplemented with 2.0 mg/l BAP + 1.0 mg/l IAA was found to be most prolific combination of the treatments with regard to number and length of shoots. Creamish friable compact callus accompanying multiple shoots (8.0 was achieved from nodal segment on MS medium fortified with 2.0 mg/l BAP + 0.5 mg/l 2,4-D within 8 days of culture. The in vitro regenerated shoots were rooted best in half strength MS medium enriched with 1.0 mg/l IAA. The rooted plantlets were successfully established with 60% survival. Besides that the biochemical parameters, like chlorophyll, total sugars, reducing sugars and proteins were estimated in leaf tissue from both in vivo and in vitro raised plants in order to establish the sustainability of plants.

  10. AN ANALYSIS OF THE GAPS IN THE NEWLY ESTABLISHED SOUTH AFRICAN MILITARY OMBUD

    Directory of Open Access Journals (Sweden)

    Boitumelo Mmusinyane

    2013-06-01

    Full Text Available The South African Department of Defence and Military Veterans can be commended for having taken a bold step in an endeavour to establish an independent entity capable of conducting oversight of its military through the introduction of the Military Ombud Act. However, said Act seems not to adequately address pertinent issues experienced by the defence sector. These issues include who may submit a complaint, the independence of the Military Ombud (MO and its accountability structure. Unless the Bill deals with these issues, we are likely to see dispossession of the public protector’s investigation powers and the establishment of a mere toothless tiger. Under the current format of the Bill, the MO is likely to become the Minister’s mouthpiece. It would deepen and marginalise military complainants’ hope of finding a remedy in an independent structure capable of challenging some of the questionable military acts or omissions that have no substance while not achieving the exercise, enjoyment and fulfilment of military complainants’ human rights in accordance with the 1996 Constitution of the Republic of South Africa.

  11. Gene mutations analysis in resistant Mycobacterium tuberculosis isolates%耐药结核分枝杆菌基因突变分析

    Institute of Scientific and Technical Information of China (English)

    刘家云; 苏明权; 陈超扬; 郝晓柯; 徐修礼; 孙惠平; 龙铟; 钱妙玲; 张鹏亮; 樊新; 程晓东; 马越云

    2010-01-01

    Objective To investigate the relationship between the phenotypes and the patterns of genetic mutations in the corresponding resistance genes (rpoB, katG, inhA, ahpC, rrs, rpsL, embB and gyrA) in resistant Mycobacterium tuberculosis (MTB) isolates. Methods Rifampicin-resistant gene (rpoB), isoniazid-resistant genes (katG, inhA, ahpC), streptomycin-resistant genes (rrs, rpsL), ethambutol-resistant gene (embB) and quinolinone-resistant gene (gyrA) were amplified by PCR with sequence-specific primers, then mutants screened by single-stranded conformation polymorphism (SSCP) were sequenced. Results rpoB mutation with predominant Ser450Trp pattern was 94. 9% (56/59) in 59 rifampicin-resistant isolates;katG mutation rate was 38. 9% (35/90) and the main pattern was Ser315Thr, but only 3 inhA mutants and no ahpC mutation were determined in 90 isoniazid-resistant isolates;gyrA mutation with main Asp94Gly then Ala90Val pattern was 82.4% (28/34) in 34 quinolinone-resistant isolates;the total mutation rate was 77.4% in 31 streptomycin-resistant isolates, of which 15 isolates mutated in rrs with main pattern A514C or A1041G, 10 isolates mutated in rpsL Lys88Arg;and embB mutation with main Met306Val accounted for 19.4% (6/31) in 31 ethambutol-resistant isolates. Conclusions The results showed that resistance of resistant MTB may be complicated, and DNA sequencing-based mutation analysis could efficiently detect the molecular makers such as rpoB, katG, gyrA, rrs, rpsL and embB in resistant MTB isolates. Meanwhile, it is notable that the rpoB mutation pattern in our isolates is different from previous report, further effort are needed to confirm the characteristics. The spectrum of potential resistance-related mutations in MTB clinical isolates may lay substantial foundation for the rapid molecular diagnosis and rational use of drug to MTB patients.%目的 探讨结核分枝杆菌耐药表型与基因突变位点之间的相互关系.方法 采用序列特异性引物分别扩增92

  12. BOOTSTRAPPING AND MONTE CARLO METHODS OF POWER ANALYSIS USED TO ESTABLISH CONDITION CATEGORIES FOR BIOTIC INDICES

    Science.gov (United States)

    Biotic indices have been used ot assess biological condition by dividing index scores into condition categories. Historically the number of categories has been based on professional judgement. Alternatively, statistical methods such as power analysis can be used to determine the ...

  13. A strategic analysis of future growth options for an established process control company

    OpenAIRE

    Levesque, Sheila Lynne

    2007-01-01

    This project speaks to the prevailing business environment presently encountered at WESTCOAST Controls Ltd (WESTCOAST), a leading process control company in British Columbia, Canada. The scope of the project covers topics such as company overview, external industry analysis, and internal company analysis including strategic tools such as Porter's 5 Forces. The project concludes with a recommendation for the restructuring of the control systems & solutions division for improved performan...

  14. CRAB3: Establishing a new generation of services for distributed analysis at CMS

    Science.gov (United States)

    Cinquilli, M.; Spiga, D.; Grandi, C.; Hernàndez, J. M.; Konstantinov, P.; Mascheroni, M.; Riahi, H.; Vaandering, E.

    2012-12-01

    In CMS Computing the highest priorities for analysis tools are the improvement of the end users’ ability to produce and publish reliable samples and analysis results as well as a transition to a sustainable development and operations model. To achieve these goals CMS decided to incorporate analysis processing into the same framework as data and simulation processing. This strategy foresees that all workload tools (TierO, Tier1, production, analysis) share a common core with long term maintainability as well as the standardization of the operator interfaces. The re-engineered analysis workload manager, called CRAB3, makes use of newer technologies, such as RESTFul based web services and NoSQL Databases, aiming to increase the scalability and reliability of the system. As opposed to CRAB2, in CRAB3 all work is centrally injected and managed in a global queue. A pool of agents, which can be geographically distributed, consumes work from the central services serving the user tasks. The new architecture of CRAB substantially changes the deployment model and operations activities. In this paper we present the implementation of CRAB3, emphasizing how the new architecture improves the workflow automation and simplifies maintainability. In particular, we will highlight the impact of the new design on daily operations.

  15. Establishing guidance for the review of human reliability analysis in PSA

    International Nuclear Information System (INIS)

    PSI was commissioned to develop Guidelines for the Regulatory Review of the Human Reliability Analysis (HRA) within Probabilistic Safety Assessments (PSAs) for nuclear power plants. In the Guidelines, HRA quality is addressed in terms of 97 indicators. Each indicator is formulated as a question, described as a specific feature of the analysis, and then explained in detail. Two analysis stages are distinguished: the selection of the human errors to be modelled, and their quantification to determine their impact on the core damage frequency. Review findings are grouped under two headings: transparency and adequacy. An analysis is 'transparent' if an externally qualified person is able to reproduce the analysis results, and 'adequate' if such results reflect the plant-specific conditions related to safety. To allocate resources efficiently, the review is structured in two phases: (1) The Quick Review, which clarifies whether the HRA has a fundamental deficiency and, furthermore, if it points to information needs and areas of emphasis for the detailed review, and (2) The Detailed Review, which results in well-grounded findings, based on extended examinations and close-plant contacts. (authors)

  16. COST ANALYSIS OF LONG ESTABLISHED AND NEWER ORAL ANTIEPILEPTIC DRUGS AVAILABLE IN THE INDIAN MARKET

    Directory of Open Access Journals (Sweden)

    Phatak Abhishek M, Hotwani Jitendra H, Deshmukhkiran R, Panchal Sagar S, Naik Madhura S

    2015-10-01

    Full Text Available Background: Large number of pharmaceutical companies manufactures antiepileptic drugs in India. The price variations among the marketed drugs are wide. Aims: The present study was aimed to find the cost of different oral antiepileptic drugs available in Indian market as monotherapy, combination therapy and number of manufacturing companies for each, to evaluate difference in cost of different brands of same dosage of same active drug by calculating percentage variation of cost. Methods and Materials: Cost of a drug being manufactured by different companies, in the same strength and dosage forms was obtained from “Indian Drug Review” Vol. XXI, Issue No.4, 2014 and “Current Index of Medical Specialties” July-October 2014. The difference in the maximum and minimum price of the same drug manufactured by different pharmaceutical companies and percentage variation in price was calculated. Results: The percentage price variation noted of long-established drugs was – Phenytoin (50mg: 140%, Carbamazepine (100mg: 1033%, Phenobarbital (30mg : 730%, Valproic acid (300mg : 420%. Newer drugs –Levetiracetam (250mg: 75%, Lamotrigine (25mg: 66%, Topiramate (50mg: 108%, Zonisamide (100mg: 19%. Combination drugs – Phenobarbital + Phenytoin (30+100 mg: 354.55%. Conclusion: The percentage price variation of different brands of the same commonly used long-established oral antiepileptic drug manufactured in India is very wide. The formulation or brand of Antiepileptic drugs (AED’s should preferably not be changed since variations in bioavailability or different pharmacokinetic profiles may increase the potential for reduced effect or excessive side effects. Hence, manufacturing companies should aim to decrease the price variation while maintaining the therapeutic efficacy.

  17. Use of benefit-cost analysis in establishing Federal radiation protection standards: a review

    International Nuclear Information System (INIS)

    This paper complements other work which has evaluated the cost impacts of radiation standards on the nuclear industry. It focuses on the approaches to valuation of the health and safety benefits of radiation standards and the actual and appropriate processes of benefit-cost comparison. A brief historical review of the rationale(s) for the levels of radiation standards prior to 1970 is given. The Nuclear Regulatory Commission (NRC) established numerical design objectives for light water reactors (LWRs). The process of establishing these numerical design criteria below the radiation protection standards set in 10 CFR 20 is reviewed. EPA's 40 CFR 190 environmental standards for the uranium fuel cycle have lower values than NRC's radiation protection standards in 10 CFR 20. The task of allocating EPA's 40 CFR 190 standards to the various portions of the fuel cycle was left to the implementing agency, NRC. So whether or not EPA's standards for the uranium fuel cycle are more stringent for LWRs than NRC's numerical design objectives depends on how EPA's standards are implemented by NRC. In setting the numerical levels in Appendix I to 10 CFR 50 and 40 CFR 190 NRC and EPA, respectively, focused on the costs of compliance with various levels of radiation control. A major portion of the paper is devoted to a review and critique of the available methods for valuing health and safety benefits. All current approaches try to estimate a constant value of life and use this to vaue the expected number of lives saved. This paper argues that it is more appropriate to seek a value of a reduction in risks to health and life that varies with the extent of these risks. Additional research to do this is recommended

  18. Use of benefit-cost analysis in establishing Federal radiation protection standards: a review

    Energy Technology Data Exchange (ETDEWEB)

    Erickson, L.E.

    1979-10-01

    This paper complements other work which has evaluated the cost impacts of radiation standards on the nuclear industry. It focuses on the approaches to valuation of the health and safety benefits of radiation standards and the actual and appropriate processes of benefit-cost comparison. A brief historical review of the rationale(s) for the levels of radiation standards prior to 1970 is given. The Nuclear Regulatory Commission (NRC) established numerical design objectives for light water reactors (LWRs). The process of establishing these numerical design criteria below the radiation protection standards set in 10 CFR 20 is reviewed. EPA's 40 CFR 190 environmental standards for the uranium fuel cycle have lower values than NRC's radiation protection standards in 10 CFR 20. The task of allocating EPA's 40 CFR 190 standards to the various portions of the fuel cycle was left to the implementing agency, NRC. So whether or not EPA's standards for the uranium fuel cycle are more stringent for LWRs than NRC's numerical design objectives depends on how EPA's standards are implemented by NRC. In setting the numerical levels in Appendix I to 10 CFR 50 and 40 CFR 190 NRC and EPA, respectively, focused on the costs of compliance with various levels of radiation control. A major portion of the paper is devoted to a review and critique of the available methods for valuing health and safety benefits. All current approaches try to estimate a constant value of life and use this to vaue the expected number of lives saved. This paper argues that it is more appropriate to seek a value of a reduction in risks to health and life that varies with the extent of these risks. Additional research to do this is recommended. (DC)

  19. Establishment of animal model for the analysis of cancer cell metastasis during radiotherapy

    International Nuclear Information System (INIS)

    Γ-Ionizing radiation (IR) therapy is one of major therapeutic tools in cancer treatment. Nevertheless, γ-IR therapy failed due to occurrence of metastasis, which constitutes a significant obstacle in cancer treatment. The main aim of this investigation was to construct animal model which present metastasis during radiotherapy in a mouse system in vivo and establishes the molecular mechanisms involved. The C6L transfectant cell line expressing firefly luciferase (fLuc) was treated with γ-IR, followed by immunoblotting, zymography and invasion assay in vitro. We additionally employed the C6L transfectant cell line to construct xenografts in nude mice, which were irradiated with γ-IR. Irradiated xenograft-containing mice were analyzed via survival curves, measurement of tumor size, and bioluminescence imaging in vivo and ex vivo. Metastatic lesions in organs of mice were further assessed using RT-PCR, H & E staining and immunohistochemistry. γ-IR treatment of C6L cells induced epithelial-mesenchymal transition (EMT) and increased cell invasion. In irradiated xenograft-containing mice, tumor sizes were decreased dramatically and survival rates extended. Almost all non-irradiated xenograft-containing control mice had died within 4 weeks. However, we also observed luminescence signals in about 22.5% of γ-IR-treated mice. Intestines or lungs of mice displaying luminescence signals contained several lesions, which expressed the fLuc gene and presented histological features of cancer tissues as well as expression of EMT markers. These findings collectively indicate that occurrences of metastases during γ-IR treatment accompanied induction of EMT markers, including increased MMP activity. Establishment of a murine metastasis model during γ-IR treatment should aid in drug development against cancer metastasis and increase our understanding of the mechanisms underlying the metastatic process

  20. Establishment of Orthotopic Xuanwei Lung Cancer SCID Mouse Model 
and Analysis of Biological Properties

    Directory of Open Access Journals (Sweden)

    Yongchun ZHOU

    2012-08-01

    Full Text Available Background and objective The incidence of Xuanwei lung cancer ranks first in China, and its pathogenesis requires in-depth investigation. This study aims to establish an orthotopic Xuanwei lung cancer severe combined immunodeficiency (SCID mouse model and to provide a basic experimental platform for further study. Methods The Xuanwei lung cancer cell line XWLC-05 was inoculated into the lung tissue of SCID mice in high and low doses. The tumor formation rates, tumor characteristics, spontaneous metastases, and survival times of the mice were observed, taking a subcutaneously transplanted tumor as control. Results The tumor formation rates of the orthotopic transplantation of lung cancer cells in high and low doses were 81% and 83%, respectively, among which mice in the high-dose group appeared cachectic on day 13. Extensive invasion and adhesion were observed in the contralateral lung and thoracic cavity, but no distant metastasis was exhibited. Mice with low-dose cells in the orthotopic transplantation group appeared cachectic and distant metastasis occurred on day 25. The tumor formation rates in the subcutaneous inoculation group by the high and low doses of cells were 100% and 94.5%, respectively, and no distant metastasis was observed. The rate of metastasis within the orthotopic transplantation group and between the orthotopic and subcutaneous inoculation groups showed a significant difference (P<0.05. A significant difference was indicated by the survival rate within and between the groups (P<0.001. Conclusion We successfully established an orthotopic XWLC SCID mouse model, which lays the foundation for a more in-depth study.

  1. ESTABLISHING DESIGNED FINANCIAL CONTROL: AN EMPIRICAL STUDY ON URBAN LOCAL BODIES IN INDIA USING FACTOR AND CLUSTER ANALYSIS

    OpenAIRE

    Sidhakam BHATTACHARYYA; Bandyopadhyay, Gautam

    2013-01-01

    We have established in earlier study that our designed financial controls help the Indian ULBs (Urban Local Bodies) to increase recurrent surplus. Our further research, as described in this paper, reveals that the concept of designed controls conforms to the result of statistical techniques used. Nonparametric tests applied on the dataset allow us to proceed for application of multivariate techniques. Factor analysis divides the variables of revenue income and expenditure into two factors whi...

  2. Disinfectant Susceptibility of Mycobacterium avium

    OpenAIRE

    Taylor, Robert Henry

    1998-01-01

    Mycobacterium avium, an opportunistic human pathogen, infects between 25 and 50% of advanced-stage acquired immuno-deficiency syndrome (AIDS) patients in the United States. M. avium has been isolated from many environmental sources including: natural waters, soils, and aerosols. M. avium has also been recovered from within municipal and hospital drinking water systems. Rhesus macaques (Macaca mulatta) infected with the simian HIV analog, SIV, have been shown to acquire M. avium infections...

  3. Growth of Mycobacterium tuberculosis Biofilms

    OpenAIRE

    Kulka, Kathleen; Hatfull, Graham; Ojha, Anil K.

    2012-01-01

    Mycobacterium tuberculosis, the etiologic agent of human tuberculosis, has an extraordinary ability to survive against environmental stresses including antibiotics. Although stress tolerance of M. tuberculosis is one of the likely contributors to the 6-month long chemotherapy of tuberculosis 1, the molecular mechanisms underlying this characteristic phenotype of the pathogen remain unclear. Many microbial species have evolved to survive in stressful environments by self-assembling in highly o...

  4. Analysis of human protein replacement stable cell lines established using snoMEN-PR vector.

    Directory of Open Access Journals (Sweden)

    Motoharu Ono

    Full Text Available The study of the function of many human proteins is often hampered by technical limitations, such as cytotoxicity and phenotypes that result from overexpression of the protein of interest together with the endogenous version. Here we present the snoMEN (snoRNA Modulator of gene ExpressioN vector technology for generating stable cell lines where expression of the endogenous protein can be reduced and replaced by an exogenous protein, such as a fluorescent protein (FP-tagged version. SnoMEN are snoRNAs engineered to contain complementary sequences that can promote knock-down of targeted RNAs. We have established and characterised two such partial protein replacement human cell lines (snoMEN-PR. Quantitative mass spectrometry was used to analyse the specificity of knock-down and replacement at the protein level and also showed an increased pull-down efficiency of protein complexes containing exogenous, tagged proteins in the protein replacement cell lines, as compared with conventional co-expression strategies. The snoMEN approach facilitates the study of mammalian proteins, particularly those that have so far been difficult to investigate by exogenous expression and has wide applications in basic and applied gene-expression research.

  5. Analysis of Borrelia burgdorferi surface proteins as determinants in establishing host cell interactions

    Directory of Open Access Journals (Sweden)

    Virginia L Schmit

    2011-07-01

    Full Text Available Borrelia burgdorferi infection causes Lyme borreliosis in humans, a condition which can involve a systemic spread of the organism to colonize various tissues and organs. If the infection is left untreated by antimicrobials, it can lead to manifestations including, arthritis, carditis, and/or neurological problems. Identification and characterization of B. burgdorferi outer membrane proteins that facilitate cellular attachment and invasion to establish infection continue to be investigated. In this study, we sought to further define putative cell binding properties of surface-exposed B. burgdorferi proteins by observing whether cellular adherence could be blocked by antibodies. B. burgdorferi mixed separately with monoclonal antibodies against outer surface protein (Osp A, OspC, decorin-binding protein (Dbp A, BBA64, and RevA antigens were incubated with human umbilical vein endothelial cells (HUVEC and human neuroglial cells (H4. B. burgdorferi treated with anti-OspA, -DbpA, and –BBA64 monoclonal antibodies showed a significant decrease in cellular association compared to controls, whereas B. burgdorferi treated with anti-OspC and anti-RevA showed no reduction in cellular attachment. Additionally, temporal transcriptional analyses revealed upregulated expression of bba64, ospA, and dbpA during coincubation with cells. Together, the data provide evidence that OspA, DbpA, and BBA64 function in host cell adherence and infection mechanisms.

  6. Establishment of microsatellite-based triplex PCR for parentage analysis of Chinese shrimp Fenneropenaeus chinensis

    Institute of Scientific and Technical Information of China (English)

    GAO Huan; KONG Jie; LIU Ping; MENG Xianhong; LUAN Sheng; ZHANG Tianshi

    2007-01-01

    Through exploring the microsatellite primers from the random genome sequences of Chinese shrimp (Fenneropenaeus chinensis), some microsatellite primers were obtained with rich polymorphic genetic information, and a triplex PCR was established using three primers (RS1101, RS0683 and H081 primers). By adjusting the final concentration of Mg2+, dNTP and primers, and using a touch-town PCR program, the optimum amplification parameters of PCR system were obtained, which could successfully amplify the three primers in a PCR reaction. In the denatured PAGE gel, the amplified DNA fragments of three primers RS1 101,RS0683 and H081 could be easily identified each other. For the triplex PCR system, the PPE (probabilities of paternity exclusion) is 0.967 9,and the DP (discrimination power) is 0.999 327.Using the triplex PCR to test ten individuals of a parentage and their parents, an individual was excluded from the parentage in all of the three microsatellite loci, which might be mixed into the parentage for some unknown reason such as factitious misplay. The triplex PCR will be of great practical value in identifying the parentages of F. chinensis.

  7. Establishment of Comprehensive Evaluation Indicator System for Sustainable Development of Microcredit and Empirical Analysis

    Institute of Scientific and Technical Information of China (English)

    YANG Di-hang; LUO He-hua

    2012-01-01

    We establish the comprehensive evaluation indicator system for sustainable development of micro-credit,constituted by the profitability ratio,financial constitution ratio,loan quality ratio and operating efficiency ratio.Taking the case of micro-credit of China Foundation for Poverty Alleviation,we measure the current situation of sustainable development of micro-credit.As far as we are concerned,the composite index of sustainable development of micro-credit shows the growth trend on the whole.Finally,in order to further improve the sustainable development of microcredit,provide farmers with better financial services and enable more farmers to benefit from micro-credit,we put forth the following recommendations:strengthening technological innovation to reduce costs and improve operating efficiency and profitability of the micro-credit;strengthening risk control to improve the quality of loans;providing all-around non-financial services to lay the foundation for the sustainable development of microcredit;setting an appropriate level of interest rates to improve the profitability of the institutions.

  8. Establishing a Link Between Prescription Drug Abuse and Illicit Online Pharmacies: Analysis of Twitter Data

    OpenAIRE

    Katsuki, Takeo; Mackey, Tim Ken; Cuomo, Raphael

    2015-01-01

    Background Youth and adolescent non-medical use of prescription medications (NUPM) has become a national epidemic. However, little is known about the association between promotion of NUPM behavior and access via the popular social media microblogging site, Twitter, which is currently used by a third of all teens. Objective In order to better assess NUPM behavior online, this study conducts surveillance and analysis of Twitter data to characterize the frequency of NUPM-related tweets and also ...

  9. The Process of Wage Adjustment: An Analysis Using Establishment-Level Data

    OpenAIRE

    Bayo-Moriones, Alberto; Galdón-Sánchez, José Enrique; Martinez-de-Morentin, Sara

    2011-01-01

    This article presents a study of the influences on the factors that shape wage adjustments. The cost of living, comparability with other firms' wages, the fulfilment of collective agreements at sector level, the need to recruit and retain employees, the performance of the organisation, and the climate of industrial relations are included as factors of interest. The analysis was carried out using a sample of Spanish manufacturing plants. Our results show that the structural characteristics of ...

  10. Basic principles for establishing best practice materials characterisation methods based on Rietveld diffraction analysis

    International Nuclear Information System (INIS)

    The effectiveness of Rietveld modelling of materials microstructure character is often limited by practitioners being unfamiliar with the basis of non-linear least squares analysis and its limitations. This paper considers the basis of good practice with particular reference to (1) the critical importance of data quality and (2) careful management of the refinement process. The analytical descriptors considered are phase composition levels and lattice parameters. (Author)

  11. Road Incidents Within an Georeferenced Area Analysis to Establish Odds of Hospitalization. An Application

    Directory of Open Access Journals (Sweden)

    Fernanda Martínez Micakoski

    2015-07-01

    Full Text Available The association analysis identifies the existence of a relationship between two or more variables, this usinga statistical test allows determining the influence between an attribute measured in a traffic event and thehospitalization of the participant, allowing obtaining patterns related the seriousness of the incident in connection totheir need for health care. Since the causes and consequences of the incident differ according to the sector beingstudied, in this case Trenque Lauquen Provincia de Buenos Aires of Argentina, city of about 40,000 habitants, themethodology developed use a grouping in zones and a grouping of measurements of each incident in responseto the changes of Humans, Vehicles and Environments factors. The application of statistical analysis is validatedbased on the application of the model with data not used in the analysis. The results in all cases allow concludedthat the model provides accurate information for make a diagnosis or assess an intervention. The present studyprovides the characterization of each zone according to their most critical result, which is the health impact onthe person involved. Based on this description it is possible to tailor preventive strategies efficiently and link thespending with simple evaluation programs.

  12. ESTABLISHMENT OF A HUMAN T-LYMPHOMA CELL LINE(H-TL90) AND ANALYSIS OF ITS BIOLOGICAL CHARACTERISTICS

    Institute of Scientific and Technical Information of China (English)

    史历; 刘旭; 张月梅; 李有芳; 李殿俊; 王吾如

    1995-01-01

    We established a human T-lymphoma cell line from the cancerous ascites of a male patient with prostate cancer which was named H-TL90. This cell line was characterized by its histological features, and by chromosomal and immunological analysis. Immunophenotypic analysis revealed that the cells expressed surface antigen CD3- CD4- CD7+ CD8-. Biological analysis revealed that the cell can promote lymphocyte proliferation. This suggested that the cell line has an autosecretion function. Cytogenetic analysis revealed that H-TL90 was a hyperdlploid with 47 chromosomes and had characteristic translocation between chromosome 3 and 11, and the deletion of the long arm of chromosome 6. These results demonstrated the H-TL90 cell line can be a useful modal for the study of human T-lymphoma.

  13. Cloning, purification, crystallization and preliminary X-ray analysis of ESX-1-secreted protein regulator (EspR) from Mycobacterium tuberculosis

    International Nuclear Information System (INIS)

    ESX-1 secreted protein regulator (EspR, Rv3849) from M. tuberculosis has been purified and crystallized, and diffracted to 3.2 Å resolution at wavelength 0.97625 Å. ESX-1-secreted protein regulator (EspR; Rv3849) is a key regulator in Mycobacterium tuberculosis that delivers bacterial proteins into the host cell during infection. EspR binds directly to the Rv3616c-Rv3614c promoter and activates transcription and secretes itself from the bacterial cell by the ESX-1 system. The three-dimensional structure of EspR will aid in understanding the mechanisms by which it binds to the Rv3616c-Rv3614c promoter and is involved in transcriptional activation. This study will significantly aid in the development of EspR-based therapeutics against M. tuberculosis. The full-length EspR gene from M. tuberculosis (H37Rv strain) was cloned and overexpressed as a soluble protein in Escherichia coli. The protein was purified by affinity chromatography using His-tagged protein followed by size-exclusion chromatography. EspR was crystallized using polyethylene glycol 3350 as precipitant. The crystals diffracted to 3.2 Å resolution using synchrotron radiation of wavelength 0.97625 Å. The crystal belonged to space group P3121 and contained three monomers in the asymmetric unit. Native and heavy-atom-derivatized data sets were collected from EspR crystals for use in ab initio structure-solution techniques

  14. Structural analysis of sigma E interactions with core RNA polymerase and its cognate P-hsp20 promoter of Mycobacterium tuberculosis.

    Science.gov (United States)

    Gupta, Aayatti Mallick; Pal, Purab; Mandal, Sukhendu

    2016-04-01

    Alternate sigma factor plays an important role for the survival of Mycobacterium tuberculosis in adverse environmental condition. Stress-induced sigma factors are major cause for expression of genes involved in pathogenesis, dormancy and various unusual environmental conditions. In the present work, an attempt has been made to characterize one of such M. tuberculosis (Mtb) sigma factor, SigE. The structures of Mtb-SigE and Mtb-β have been predicted using comparative modelling techniques and validated. Effort has also been implied to understand the nature of interaction of SigE with the core RNA polymerase subunits which have well identified the amino acid residues in the binding interface and prompted the fact that Mtb-β' and Mtb-β interact with domain 2 and domain 4 of Mtb-SigE, respectively. Furthermore, intermolecular docking study predicted the interface between the Mtb-SigE and its putative promoter P-hsp20. The report confers the probable amino acid residues and the nitrogenous bases involved in the recognition of P-hsp20 by the sigma factor to initiate the transcription process. PMID:26006066

  15. Cost Analysis of Establishing a Relationship Between a Surgical Program in the US and Vietnam

    OpenAIRE

    Quyen D. Chu; Nguyen, Thu; Nguyen, Phuong; Ho, Hung S.

    2012-01-01

    “Twinning” refers to a constructive partnership between hospitals in developed and developing nations. Such an effort may contribute immensely to capacity building for the developing nation, but one of the reasons given for the lack of sustainability is cost. We share a detailed operating cost analysis of our recent experience with an institution in Vietnam. We were awarded a 1-year $54,000 grant from the Vietnam Education Foundation (VEF) to conduct a live tele-video conferencing course on t...

  16. Establishment of a Risk Assessment Framework for Analysis of the Spread of Highly Pathogenic Avian Influenza

    Institute of Scientific and Technical Information of China (English)

    LI Jing; WANG Jing-fei; WU Chun-yan; YANG Yan-tao; JI Zeng-tao; WANG Hong-bin

    2007-01-01

    To evaluate the risk of highly pathogenic avian influenza (HPAI) in mainland China, a risk assessment framework was built.Risk factors were determined by analyzing the epidemic data using the brainstorming method; the analytic hierarchy process was designed to weigh risk factors, and the integrated multicriteria analysis was used to evaluate the final result.The completed framework included the risk factor system, data standards for risk factors, weights of risk factors, and integrated assessment methods. This risk assessment framework can be used to quantitatively analyze the outbreak and spread of HPAI in mainland China.

  17. Establishment and validation of a dose-effect curve for {gamma}-rays by cytogenetic analysis

    Energy Technology Data Exchange (ETDEWEB)

    Barquinero, Joan F.; Caballin, Maria Rosa [Unitat d`Antropologia, Departament de Biologia Animal, Biologia Vegetal i Ecologia, Facultat de Ciencies, Universitat Autonoma de Barcelona, Bellaterra (Spain); Barrios, Leonardo; Ribas, Montserrat [Unitat de Biologia Cel.lular, Departament de Biologia Cel.lular i Fisiologia, Facultat de Ciencies, Universitat Autonoma de Barcelona, Bellaterra (Spain); Miro, Rosa [Institut de Biologia Fondamental `Vicent Villar Palasi`, Universitat Autonoma de Barcelona, Barcelona (Spain); Egozcue, Josep [Servei d`Oncologia, Hospital de la Santa Crue i Sant Pau, Universitat Autonome de Barcelona, Barcelona (Spain)

    1995-01-01

    A dose-effect curve obtained by analysis of dicentric chromosomes after irradiation of peripheral blood samples, from one donor, at 11 different doses of {gamma}-rays is presented. For the elaboration of this curve, more than 18,000 first division metaphases have been analyzed. The results fit very well to the linear-quadratic model. To validate the curve, samples from six individuals (three controls and three occupationally exposed persons) were irradiated at 2 Gy. The results obtained, when compared with the curve, showed that in all cases the 95% confidence interval included the 2 Gy dose, with estimated dose ranges from 1.82 to 2.19 Gy.

  18. Analysis of Breast Thermography Using Fractal Dimension to Establish Possible Difference between Malignant and Benign Patterns

    Directory of Open Access Journals (Sweden)

    Mahnaz Etehad Tavakol

    2010-01-01

    Full Text Available Early detection of breast cancer by means of thermal imaging has a long and extremely controversial history. Recently, the availability of highly sensitive infrared (IR cameras which can produce high-resolution diagnostic images of the temperature and vascular changes of breasts, as well as a better knowledge of advanced image processing techniques, has generated a renewed interest. The objective of this study is to investigate fractal analysis of breast thermal images and to develop an algorithm for detecting benignity and malignancy of breast diseases. The study is based on IR images captured by thermal camera, in which the resolution of the results is within the state of the art of IR camera. A total of 7 malignant cases and 8 benign cases have been considered. The breast images were first segmented by fuzzy c-means clustering. Then the first hottest regions for each image were identified and the fractal dimension of those regions was computed. It is shown that the fractal dimension results significantly differ between malignant and benign patterns, suggesting that fractal analysis may potentially improve the reliability of thermography in breast tumor detection.

  19. The Analysis of the Relationship between Communication Skills and the Establishment of Clark's Management Network among Sport Managers

    Directory of Open Access Journals (Sweden)

    Rasool NAZARI

    2016-03-01

    Full Text Available The purpose of this study was to analyze the relationship between communication skills and the establishment of Clark's Management Network among sport managers. This applied research is a kind of correlational – survey study. Statistical population consist ed of 140 sport managers working for sport organizations in Isfahan Province, which have been selected by stratified random sampling. Measurement tools included two questionnaires of Communication Skill (91% and Establishment of Clark's Management Network (87%. Descriptive and inferential statistical methods applied for statistical analysis using SPSS software. Results show that there is a relationship between communication skills and the establishment of Clark's Management Network among sport managers. I n addition, the feedback component has greater contribution to prediction of Clark's management network (p 0.05. In general, it can acknowledge that the communication skills are effective in the establishment of Clark's management network and cause to i mprove the management process of sport organizations' managers and to achieve to the organizational goals with high level of productivity.

  20. Establishing RNA interference as a reverse-genetic approach for gene functional analysis in protoplasts.

    Science.gov (United States)

    Zhai, Zhiyang; Sooksa-nguan, Thanwalee; Vatamaniuk, Olena K

    2009-02-01

    Double-stranded (ds)RNA interference (RNAi) is widely used for functional analysis of plant genes and is achieved via generating stable transformants expressing dsRNA in planta. This study demonstrated that RNAi can also be utilized to examine gene functions in protoplasts. Because protoplasts are nongrowing cells, effective RNAi-triggered gene silencing depends not only on a depletion of gene transcripts but also on turnover rates of corresponding polypeptides. Herein, we tested if transient RNAi in protoplasts would result in the depletion of a targeted polypeptide and, because protoplasts have a limited life span, if functional assays of RNAi knockout genes would be feasible in protoplasts. We showed that protoplasts transfection with an in vitro-synthesized dsRNA against Arabidopsis (Arabidopsis thaliana) beta-glutamylcysteine synthase (ECS1), a key enzyme in the synthesis of glutathione, resulted in a 95% depletion of ECS1 transcript, a 72% decrease of ECS1 polypeptide, and a 60% drop in glutathione content. These results were comparable with those obtained upon analysis of Arabidopsis seedlings bearing the cad2-1 mutant allele of ECS1. We also improved the procedure for RNAi inactivation of several genes simultaneously. Finally, because we isolated protoplasts from tissues of 14-d-old seedlings instead of 1-month-old mature plants, the described procedure is rapid (as it only takes 20 d from seed planting to functional studies), suitable for analyzing multiple genes in parallel, and independent of cloning dsRNAs into plant expression vectors. Therefore, RNAi in protoplasts complements existing genetic tools, as it allows rapid, cost- and space-efficient initial screening and selection of genes for subsequent in planta studies.

  1. Establishing the date of Maori environmental impact in New Zealand through pollen analysis and radiocarbon dating

    International Nuclear Information System (INIS)

    Full text: Over the last decade there has been an intense debate about whether New Zealand prehistory is long ( > 1500 years) or short ( < 800 years). Pollen and charcoal analyses have played a key role in this debate by helping to pinpoint the transition from relatively undisturbed environments to those deforested by anthropogenic fires. Problems with in situ contamination, reworking of sediments, confusion of natural with anthropogenic impacts, and different theoretical expectations of growth, spread and impact of early Maori populations have led to disparate conclusions. We review pollen based studies carried out on a variety of fossil sites, including peat bogs, swamps, estuaries and lakes, and contribute new results. Different sedimentary environments show varying susceptibilities to contamination and have resulted in a wide spread of ages for initial Maori impact. Datable materials least susceptible to contamination by old or young carbon are pure peat and macrofossils, whereas lake, swamp and silty sediments are most susceptible. Analysis of the radiocarbon ages obtained for the start of Maori deforestation show that ages falling in the 'long' prehistory period are exclusively derived from lake sediments and swamps. In contrast, the bulk of the ages falling in the 'short' prehistory period are from pure peat and selected plant fragments. We conclude from our analysis of radiocarbon ages for pollen based deforestation that the first evidence of Maori environmental impact began about 700-550 calendar years BP (1250-1400 AD). Finer age resolution is limited by dating techniques, site limitations and the uncertainty associated with identifying the first signs of human impact. The period we have identified corresponds with the oldest dated archaeological sites and supports the short prehistory hypothesis. We discuss how to distinguish reliable fossil sites from those that have a high risk of giving misleading results

  2. Establishing time-dependent model of deformation modulus caused by bedrock excavation rebound by inverse analysis method

    Institute of Scientific and Technical Information of China (English)

    2008-01-01

    Rock rebound relaxation deformation,or even rock burst,caused by the excavation of dam base and abutment or high rock slope affects their stability and results in the fall of mechanical properties of the rock.So an inverse analysis method was proposed in this paper to establish the time-dependent model of deformation modulus caused by excavation rebound.The basic principle is based on the combination of observed data of the excavation rebound deformation of dam abutment or rock slope,and the calculated rebound deformation by FEM under ground stress at the corresponding time in the excavation process.The norm of the residuals of observed data and calculated data are taken as the objective function.Accordingly,the time-dependent model of bedrock deformation modulus can be established.The method displays its significance in the design of excavation,construction and operation management of dam base and high slope.

  3. Analysis of Establishment and MHD Stability of a Free Curve-Surface Flow for Liquid Metal PFCs

    International Nuclear Information System (INIS)

    Full text: An innovation concept of three layer s guidable liquid metal free curve-surface flow is addressed and its establishment and MHD stability are also analyzed on theoretically Layer I is a basic conduction layer, layer II is a key adjust layer, layer III is the surface layer. To adjust layer I and II in suitable flowing conditions, the MHD effect stability surface layer III can be obtained. In meantime, the layer I and II can be as the heat sink and the coolant flow (it is also suitable to a flat surface flow to avoid rivulet flow). According to Newton's laws and fluid mechanical principles, the analysis results show that an MHD effect stability free curve surface flow can be established under a given curve surface in a gradient magnetic field. (author)

  4. Beta-lactamases of Mycobacterium tuberculosis and Mycobacterium kansasii.

    Science.gov (United States)

    Segura, C; Salvadó, M

    1997-09-01

    Re-emergence of infectious diseases caused by mycobacteria as well as the emergence of multiresistant strains of Mycobacterium has promoted the research on the use of beta-lactames in the treatment of such diseases. Mycobacteria produce beta-lactamases: M. tuberculosis produces a wide-spectrum beta-lactamase whose behaviour mimicks those of Gram-negative bacteria. M. kansasii produces also beta-lactamase which can be inhibited by clavulanic acid. An overview on beta-lactamases from both species is reported.

  5. Inhibition of Mycobacterium tuberculosis topoisomerase I by m-AMSA, a eukaryotic type II topoisomerase poison.

    Science.gov (United States)

    Godbole, Adwait Anand; Ahmed, Wareed; Bhat, Rajeshwari Subray; Bradley, Erin K; Ekins, Sean; Nagaraja, Valakunja

    2014-04-18

    m-AMSA, an established inhibitor of eukaryotic type II topoisomerases, exerts its cidal effect by binding to the enzyme-DNA complex thus inhibiting the DNA religation step. The molecule and its analogues have been successfully used as chemotherapeutic agents against different forms of cancer. After virtual screening using a homology model of the Mycobacterium tuberculosis topoisomerase I, we identified m-AMSA as a high scoring hit. We demonstrate that m-AMSA can inhibit the DNA relaxation activity of topoisomerase I from M. tuberculosis and Mycobacterium smegmatis. In a whole cell assay, m-AMSA inhibited the growth of both the mycobacteria.

  6. Comparative proteomic analysis of sequential isolates of Mycobacterium tuberculosis from a patient with pulmonary tuberculosis turning from drug sensitive to multidrug resistant

    Directory of Open Access Journals (Sweden)

    Amit Singh

    2015-01-01

    Full Text Available Background & objectives: Tuberculosis is a major health problem in India, and the emergence of multidrug resistant (MDR and extensively drug resistant (XDR strains of Mycobacterium tuberculosis (Mtb has further complicated the situation. Though several studies characterizing drug sensitive and drug resistant strains are available in literature, almost all studies are done on unrelated strains. Therefore, the objective of this study was to compare the proteomic data of four sequential isolates of Mtb from a single patient who developed MDR-TB during the course of anti-tuberculosis therapy (ATT. Methods: In this study, using two-dimensional (2D gel electrophoresis and MALDI-TOF mass spectrometry, we compared and analyzed the cell lysate proteins of Mtb sequential clinical isolates from a patient undergoing anti-TB treatment. The mRNA expression levels of selected identified proteins were determined by quantitative real-time polymerase chain reaction (qRT-PCR. Results: The genotypes of all four isolates remained homologous, indicating no re-infection. The initial isolate (before treatment was sensitive to all first-line drugs, but the consecutive isolates were found to be resistant to isoniazid (INH and rifampicin (RIF and developed mutations in the katG, inhA and rpoB. the intensities of 27 protein spots were found to be consistently overexpressed in INH and RIF resistant isolates. The most prominent and overexpressed proteins found during the development of drug resistance were GarA (Rv1827, wag31 (Rv2145c, Rv1437 and Rv2970c. Interpretation & conclusions: This preliminary proteomic study provides an insight about the proteins that are upregulated during drug resistance development. These upregulated proteins, identified here, could prove useful as immunodiagnostic and possibly drug resistant markers in future. However, more studies are required to confirm these findings.

  7. Uncertainty analysis as essential step in the establishment of the dynamic Design Space of primary drying during freeze-drying

    DEFF Research Database (Denmark)

    Mortier, Severine Therese F. C.; Van Bockstal, Pieter-Jan; Corver, Jos;

    2016-01-01

    -drying, there are only two main process variables to be set, i.e. the shelf temperature and the chamber pressure, however preferably in a dynamic way. This manuscript focuses on the essential use of uncertainty analysis for the determination and experimental verification of the dynamic primary drying Design Space...... for pharmaceutical freeze-drying. Traditionally, the chamber pressure and shelf temperature are kept constant during primary drying, leading to less optimal process conditions. In this paper it is demonstrated how a mechanistic model of the primary drying step gives the opportunity to determine the optimal dynamic...... of the Design Space, although it is often neglected. To quantitatively assess the inherent uncertainty on the parameters of the mechanistic model, an uncertainty analysis was performed to establish the borders of the dynamic Design Space, i.e. a time-varying shelf temperature and chamber pressure, associated...

  8. Analysis of antibiotics resistance of Mycobacterium tuberculosis during 2008-2010%2008-2010年结核分枝杆菌的耐药性分析

    Institute of Scientific and Technical Information of China (English)

    王湘,; 张丽霞

    2011-01-01

    摘要:目的 了解2008-2010年结核患者分离出的结核分枝杆菌对常用抗结核药物的耐药性变迁.方法 采用罗氏培养系统进行结核分枝杆菌培养,药敏试验采用绝对浓度法,回顾性分析天津市海河医院分离的455株痰结核分枝杆菌的耐药性.结果455株结核分枝杆菌对链霉素、左氧氟沙星、利福平、乙胺丁醇、异烟肼、阿米卡星、对氨基水杨酸的总耐药率分别为29.2%、24.6%、23.1%、18.2%、18.5%、13.4%、9.2%.结论结核分枝杆菌的耐药率仍然较高,规范、足量、联合用药,是减缓耐药率的主要途径.%OBJECTIVE To investigate the variance of antimicrobial resistance and drug susceptibility of seven anti-TB drugs in hospitalized tuberculosis patients during 2008 - 2010. METHODS Roche system was used to culture Mycobacterium tuberculosis and absolute concentration method was used to test the drug susceptibility. Drug resistance and variance of antimicrobial resistance rates of 455 strains were retrospectively analyzed. RESULTS Of 455 strains, the total drug resistance rates of streptomycin, levofloxacin, rifampicin, ethambutol, isoniazid, amikacin, para-aminosalicylic acid were 29. 2%, 24. 6%, 23. 1%, 18. 2%, 18. 5%, 13. 4% and 9. 2%, respectively. CONCLUSION The resistance rate of M. Tuberculosis is still high. Standard, united, full course of treatment medication is still the main way to reduce the drug resistance.

  9. Machine Learning Model Analysis and Data Visualization with Small Molecules Tested in a Mouse Model of Mycobacterium tuberculosis Infection (2014-2015).

    Science.gov (United States)

    Ekins, Sean; Perryman, Alexander L; Clark, Alex M; Reynolds, Robert C; Freundlich, Joel S

    2016-07-25

    The renewed urgency to develop new treatments for Mycobacterium tuberculosis (Mtb) infection has resulted in large-scale phenotypic screening and thousands of new active compounds in vitro. The next challenge is to identify candidates to pursue in a mouse in vivo efficacy model as a step to predicting clinical efficacy. We previously analyzed over 70 years of this mouse in vivo efficacy data, which we used to generate and validate machine learning models. Curation of 60 additional small molecules with in vivo data published in 2014 and 2015 was undertaken to further test these models. This represents a much larger test set than for the previous models. Several computational approaches have now been applied to analyze these molecules and compare their molecular properties beyond those attempted previously. Our previous machine learning models have been updated, and a novel aspect has been added in the form of mouse liver microsomal half-life (MLM t1/2) and in vitro-based Mtb models incorporating cytotoxicity data that were used to predict in vivo activity for comparison. Our best Mtb in vivo models possess fivefold ROC values > 0.7, sensitivity > 80%, and concordance > 60%, while the best specificity value is >40%. Use of an MLM t1/2 Bayesian model affords comparable results for scoring the 60 compounds tested. Combining MLM stability and in vitro Mtb models in a novel consensus workflow in the best cases has a positive predicted value (hit rate) > 77%. Our results indicate that Bayesian models constructed with literature in vivo Mtb data generated by different laboratories in various mouse models can have predictive value and may be used alongside MLM t1/2 and in vitro-based Mtb models to assist in selecting antitubercular compounds with desirable in vivo efficacy. We demonstrate for the first time that consensus models of any kind can be used to predict in vivo activity for Mtb. In addition, we describe a new clustering method for data visualization and apply this

  10. Identification of outer membrane proteins of Mycobacterium tuberculosis.

    Science.gov (United States)

    Song, Houhui; Sandie, Reatha; Wang, Ying; Andrade-Navarro, Miguel A; Niederweis, Michael

    2008-11-01

    The cell wall of mycobacteria includes an unusual outer membrane of extremely low permeability. While Escherichia coli uses more than 60 proteins to functionalize its outer membrane, only two mycobacterial outer membrane proteins (OMPs) are known. The porin MspA of Mycobacterium smegmatis provided the proof of principle that integral mycobacterial OMPs share the beta-barrel structure, the absence of hydrophobic alpha-helices and the presence of a signal peptide with OMPs of gram-negative bacteria. These properties were exploited in a multi-step bioinformatic approach to predict OMPs of M. tuberculosis. A secondary structure analysis was performed for 587 proteins of M. tuberculosis predicted to be exported. Scores were calculated for the beta-strand content and the amphiphilicity of the beta-strands. Reference OMPs of gram-negative bacteria defined threshold values for these parameters that were met by 144 proteins of unknown function of M. tuberculosis. Two of them were verified as OMPs by a novel two-step experimental approach. Rv1698 and Rv1973 were detected only in the total membrane fraction of M. bovis BCG in Western blot experiments, while proteinase K digestion of whole cells showed the surface accessibility of these proteins. These findings established that Rv1698 and Rv1973 are indeed localized in the outer membrane and tripled the number of known OMPs of M. tuberculosis. Significantly, these results provide evidence for the usefulness of the bioinformatic approach to predict mycobacterial OMPs and indicate that M. tuberculosis likely has many OMPs with beta-barrel structure. Our findings pave the way to identify the set of proteins which functionalize the outer membrane of M. tuberculosis. PMID:18439872

  11. Mycobacterium tuberculosis: factores de virulencia

    Directory of Open Access Journals (Sweden)

    Reinier Borrero

    2011-04-01

    Full Text Available Mycobacterium tuberculosis es el agente causal de la tuberculosis, una de las enfermedades infecciosas más letales en el mundo. La única vacuna disponible para su control es el BCG, sin embargo, falla en la protección contra la tuberculosis pulmonar, siendo esta la forma más frecuente y responsable de la diseminación. La identificación de factores de virulencia del microorganismo causal pudiera ayudar en el desarrollo de un nuevo candidato vacunal que sea capaz de neutralizar la acción de esos determinantes patogénicos. El empleo de diferentes modelos animales ha permitido reproducir las etapas de la enfermedad, así como medir o cuantificar la virulencia de las distintas cepas circulantes de Mycobacterium tuberculosis. Las mutaciones génicas y otras técnicas de biología molecular han posibilitado dilucidar los genes específicos involucrados en la virulencia de este microorganismo que codifican para múltiples y complejos factores de diferente naturaleza.

  12. Five-Year Outbreak of Community- and Hospital-Acquired Mycobacterium porcinum Infections Related to Public Water Supplies ▿ †

    OpenAIRE

    Brown-Elliott, Barbara A.; Wallace, Richard J.; Tichindelean, Carmen; Sarria, Juan C.; McNulty, Steven; Vasireddy, Ravikaran; Bridge, Linda; Mayhall, C. Glenn; Turenne, Christine; Loeffelholz, Michael

    2011-01-01

    Mycobacterium porcinum is a rarely encountered rapidly growing Mycobacterium (RGM). We identified M. porcinum from 24 patients at a Galveston university hospital (University of Texas Medical Branch) over a 5-year period. M. porcinum was considered a pathogen in 11 (46%) of 24 infected patients, including 4 patients with community-acquired disease. Retrospective patient data were collected, and water samples were cultured. Molecular analysis of water isolates, clustered clinical isolates, and ...

  13. Establishment and characterization of two primary breast cancer cell lines from young Indian breast cancer patients: mutation analysis.

    Science.gov (United States)

    Pandrangi, Santhi Latha; Raju Bagadi, Sarangadhara Appala; Sinha, Navin Kumar; Kumar, Manoj; Dada, Rima; Lakhanpal, Meena; Soni, Abha; Malvia, Shreshtha; Simon, Sheeba; Chintamani, Chintamani; Mohil, Ravindar Singh; Bhatnagar, Dinesh; Saxena, Sunita

    2014-01-01

    Two novel triple negative breast cancer cell lines, NIPBC-1 and NIPBC-2 were successfully established from primary tumors of two young breast cancer patients aged 39 and 38 years respectively, diagnosed as infiltrating duct carcinoma of breast. Characterization of these cell lines showed luminal origin with expression of epithelial specific antigen and cytokeratin 18 and presence of microfilaments and secretary vesicles, microvilli, tight junctions and desmosomes on ultra-structural analysis. Both the cell lines showed anchorage independent growth and invasion of matrigel coated membranes. Karyotype analysis showed aneuploidy, deletions and multiple rearrangements in chromosomes 7, 9, X and 11 and isochromosomes 17q in both the cell lines. P53 mutational analysis revealed no mutation in the coding region in both the cell lines; however NIPBC-2 cell line showed presence of heterozygous C/G polymorphism, g.417 C > G (NM_000546.5) resulting in Arg/Pro allele at codon 72 of exon 4. Screening for mutations in BRCA1&2 genes revealed presence of three heterozygous polymorphisms in exon 11 of BRCA1 and 2 polymorphisms in exons 11, and14 of BRCA2 gene in both the cell lines. Both the cell lines showed presence of CD 44+/24-breast cancer stem cells and capability of producing mammosphere on culture. The two triple negative breast cancer cell lines established from early onset breast tumors can serve as novel invitro models to study mechanisms underlying breast tumorigenesis in younger age group patients and also identification of new therapeutic modalities targeting cancer stem cells. PMID:24502646

  14. Detection of Mycobacterium tuberculosis and Mycobacterium avium Complexes by Real-Time PCR in Bovine Milk from Brazilian Dairy Farms.

    Science.gov (United States)

    Bezerra, André Vinícius Andrade; Dos Reis, Emily Marques; Rodrigues, Rogério Oliveira; Cenci, Alexander; Cerva, Cristine; Mayer, Fabiana Quoos

    2015-05-01

    Foodborne diseases are a public health problem worldwide. The consumption of contaminated raw milk has been recognized as a major cause of transmission of bovine tuberculosis to humans. Other mycobacteria that may be present in raw milk and may cause diseases are those belonging to the Mycobacterium avium complex. In this study, molecular biology tools were applied to investigate raw milk contamination with Mycobacterium spp. in family dairy farms from Rio Grande do Sul, southern Brazil. Furthermore, different variables related to the source of the milk, herd characteristics, and management were evaluated for their effect on milk contamination. Five hundred and two samples were analyzed, of which 354 were from the Northwest region (102 farms with samples from 93 bulk tanks and 261 animals) and 148 from the South region of the state (22 farms with samples from 23 bulk tanks and 125 animals). Among them, 10 (1.99%) and 7 (1.39%) were positive for Mycobacterium tuberculosis (9 confirmed as Mycobacterium bovis) and M. avium complexes, respectively. There was no difference in the frequencies of positive samples between the regions or the sample sources. Of the positive samples, 4 were collected from a bulk tank (1 positive for M. avium and 3 for M. tuberculosis). Moreover, 1 sample was positive concomitantly for M. tuberculosis and M. avium complexes. On risk analysis, no variable was associated with raw milk contamination by M. tuberculosis complex species. However, washing the udders of all animals and drying them with paper towels were weakly classified as risk factors for M. avium contamination. Positive samples were obtained from both animals and bulk tanks, which emphasizes the importance of tuberculosis control programs and provides evidence that milk monitoring can be used as a control practice. Moreover, the findings of this study reinforce the need for awareness of the problems of raw milk consumption among the general population.

  15. Stable isotope analysis of a newly established macrofaunal food web 1.5 years after the Hebei Spirit oil spill

    International Nuclear Information System (INIS)

    Highlights: • We examined trophic structure in a newly established community after an oil spill. • This is the most extensive in situ isotopic analysis on an oiled benthic community. • Consumer-food source δ13C and δ15N rejected influx of petroleum into the community. • A novel circular statistics rejected trophic niche change of major feeding guilds. • Prevalence of omnivory and trophic plasticity may promote the recovery process. - Abstract: We examined trophic relationships in a newly established community 1.5 years after the Hebei Spirit oil spill on the west coast of Korea. Carbon and nitrogen stable isotope ratios in consumers and their potential food sources were compared between the oil-spill site and reference site, located 13.5 km from the oil-spill spot. The isotopic mixing model and a novel circular statistics rejected the influx of petrogenic carbon into the community and identified spatial consistencies such as the high contributions of microphytobenthos, food-chain length, and the isotopic niche of each feeding guild between sites. We suggested that high level of trophic plasticity and the prevalence of omnivory of consumers may promote the robustness of food web against the oil contamination. Furthermore, we highlighted the need of holistic approaches including different functional groups to quantify changes in the food web structure and assess the influence of different perturbations including oil spill

  16. New-doses limits introduction analysis for the design and operation of teletherapy facilities established by IAEA

    International Nuclear Information System (INIS)

    A design of a typical teletherapy facility was made considering a Co-60 rotating unit and using critical parameters, taking into account as a design base the dose limits established in the Safety Series No. 9 (1), and Safety Series No.115-I (2), shielding thickness when the dose limits were changed. An increment in the required thickness of 1,35 CHR for controlled areas and 2,37 CHR for non-controlled areas were found. This work considered the selection of four different types of teletherapy facilities using Co-60 sources, with different design and type of used unit. An analysis of thickness was made taking into account both the original values for the design and the real operation values in each facility. In order to determine the necessary changes for the wall thicknesses when the new recommendations are applied. (authors). 4 refs., 3 tabs

  17. ANALYSIS OF THE FORMS OF WORK AS FOR OVERCOMING DEADAPTATION MANIFESTATIONS OF THE FUTURE EDUCATORS IN HIGHER EDUCATIONAL ESTABLISHMENT

    Directory of Open Access Journals (Sweden)

    Larisa Zdanevych

    2015-04-01

    Full Text Available The author reveals theoretical approaches to the problem, interprets the views of other researchers. The problem of higher educational establishment deadaptation is cleared up in the article, interpretation of the phenomenon of teenagers’ deadaptation in different spheres of psychological-pedagogical science is given. At the modern stage deadaptation is understood as a totality of features and manifestations, which affirm inconsistency of a person’s interaction with his or her environment. And as deadaptation ruins the results, which were achieved in the process of adaptation, it is necessary in the first place to found out the essence of this phenomenon. Literature analysis and practical experience of work in the pedagogical university allow us to determine some characteristics of social status of a student of the first years of study. The analysis of different forms of pedagogical work as foe the overcoming of deadaptation manifestations of the future educators in higher educational establishment is done. The author analyses the questionering conducted at the faculty of pre-school education, which  allows to affirm that at the beginning of study students overcome educational difficulties and aren’t very thoughtful as for the perspectives of their future professional activity. In this period great responsibility is on the academic group supervisors. To prevent the manifestations of deadaptation the supervisors should should create psychological comfort in the group for the students – desirable for students state, affirming harmony in the inner, psychological and social life. To avoid the manifestations of deadaptation of the students-freshmen and in order to accelerate their adaptation, we have created the program for special course for the teachers and supervisors of academic groups.

  18. Porins Increase Copper Susceptibility of Mycobacterium tuberculosis

    OpenAIRE

    Speer, Alexander; Rowland, Jennifer L.; Haeili, Mehri; Niederweis, Michael; Wolschendorf, Frank

    2013-01-01

    Copper resistance mechanisms are crucial for many pathogenic bacteria, including Mycobacterium tuberculosis, during infection because the innate immune system utilizes copper ions to kill bacterial intruders. Despite several studies detailing responses of mycobacteria to copper, the pathways by which copper ions cross the mycobacterial cell envelope are unknown. Deletion of porin genes in Mycobacterium smegmatis leads to a severe growth defect on trace copper medium but simultaneously increas...

  19. IDENTIFICATION OF MYCOBACTERIUM GENAVENSE IN A DIANA MONKEY (CERCOPITHECUS DIANA) BY POLYMERASE CHAIN REACTION AND HIGH-PERFORMANCE LIQUID CHROMATOGRAPHY.

    Science.gov (United States)

    Kelly, Kathleen M; Wack, Allison N; Bradway, Dan; Simons, Brian W; Bronson, Ellen; Osterhout, Gerard; Parrish, Nicole M; Montali, Richard J

    2015-06-01

    A 25-yr-old Diana monkey (Cercopithecus diana) with a 1.5-yr history of chronic colitis and diarrhea was found to have disseminated granulomatous disease with intralesional acid fast bacilli. Bacilli were identified as Mycobacterium genavense by polymerase chain reaction, sequencing of the 16S-23S ribosomal RNA intergenic spacer (ITS) gene, and mycolic acid analysis by high-performance liquid chromatography. Mycobacterium genavense is a common cause of mycobacteriosis in free-ranging and captive birds. In addition, recognition of opportunistic infection in human immunodeficiency virus-positive patients is increasing. Disease manifestations of M. genavense are similar to Mycobacterium avium complex (MAC) and include fever, wasting, and diarrhea with disseminated disease. Similar clinical signs and lesions were observed in this monkey. Mycobacterium genavense should be considered as a differential for disseminated mycobacterial disease in nonhuman primates as this agent can mimic MAC and related mycobacteria.

  20. Mycobacterium tuberculosis pili (MTP), a putative biomarker for a tuberculosis diagnostic test.

    Science.gov (United States)

    Naidoo, Natasha; Ramsugit, Saiyur; Pillay, Manormoney

    2014-05-01

    Novel biomarkers are urgently needed for point of care TB diagnostics. In this study, we investigated the potential of the pilin subunit protein encoded by the mtp gene as a diagnostic biomarker. BLAST analysis of the mtp gene on published genome databases, and amplicon sequencing were performed in Mycobacterium tuberculosis Complex (MTBC) strains and other organisms. The protein secondary structure of the amino acid sequences of non-tuberculous Mycobacteria that partially aligned with the mtp sequence was analysed with PredictProtein software. The mtp gene and corresponding amino acid sequence of MTBC were 100% homologous with H37Rv, in contrast to the partial alignment of the non-tuberculous Mycobacteria. The mtp gene was present in all 91 clinical isolates of MTBC. Except for 2 strains with point mutations, the sequence was 100% conserved among the clinical strains. The mtp gene could not be amplified in all non-tuberculous Mycobacteria and respiratory organisms. The predicted MTP protein structure of Mycobacterium avium, Mycobacterium ulcerans and Mycobacterium abscessus differed significantly from that of the M. tuberculosis, which was similar to Mycobacterium marinum. The absence of the mtp gene in non-tuberculous Mycobacteria and other respiratory bacteria suggests that its encoded product, the pilin subunit protein of M. tuberculosis may be a suitable marker for a point of care TB test.

  1. Adsorbability of Mycobacterium phlei on hematite surface

    Institute of Scientific and Technical Information of China (English)

    Huifen Yang; Qiang Zhang; Zhuan Jiang

    2007-01-01

    The adsorption of microorganisms on the mineral surface is the base of microorganisms that are considered as mineral processing reagents. The principles of the use of a highly hydrophobic and negatively charged bacterium, Mycobacterium phlei, as a floc-culating-flotating agent for finely divided hematite were investigated. The flocculating-floating recovery is strongly dependent on the pH and the dosage of the bacterium. Generally the pH should be controlled over the range of 5.5-7, and the dosage should be controlled about 16 mg/L. The infrared spectrometry analysis indicates that the six functional groups of M. phlei, substituted aromatic compound groups, -(CH2)n-groups, -CH2(-CH3)groups, carbonyl groups, aromatic hydrocarbon groups, and carboxyl groups, are on the hematite surface, among which the first five ones contribute physical adsorption and only the carboxyl groups provide chemisorption. Microscopic analysis reveals that the dimensions and tight aggregation degree of the floes of hematite particles formed by M. phlei are also impacted by the pH and the content of M. phlei in flotation.

  2. Disseminated Mycobacterium tuberculosis infection in a dog.

    Science.gov (United States)

    Martinho, Anna Paula Vitirito; Franco, Marília Masello Junqueira; Ribeiro, Márcio Garcia; Perrotti, Isabella Belletti Mutt; Mangia, Simone Henriques; Megid, Jane; Vulcano, Luiz Carlos; Lara, Gustavo Henrique Batista; Santos, Adolfo Carlos Barreto; Leite, Clarice Queico Fujimura; de Carvalho Sanches, Osimar; Paes, Antonio Carlos

    2013-03-01

    An uncommon disseminated Mycobacterium tuberculosis infection is described in a 12-year-old female dog presenting with fever, dyspnea, cough, weight loss, lymphadenopathy, melena, epistaxis, and emesis. The dog had a history of close contact with its owner, who died of pulmonary tuberculosis. Radiographic examination revealed diffuse radio-opaque images in both lung lobes, diffuse visible masses in abdominal organs, and hilar and mesenteric lymphadenopathy. Bronchial washing samples and feces were negative for acid-fast organisms. Polymerase chain reaction (PCR)-based species identification of bronchial washing samples, feces, and urine revealed M. tuberculosis using PCR-restriction enzyme pattern analysis-PRA. Because of public health concerns, which were worsened by the physical condition of the dog, euthanasia of the animal was recommended. Rough and tough colonies suggestive of M. tuberculosis were observed after microbiological culture of lung, liver, spleen, heart, and lymph node fragments in Löwenstein-Jensen and Stonebrink media. The PRA analysis enabled diagnosis of M. tuberculosis strains isolated from organs. PMID:23339199

  3. Bursitis Due to Mycobacterium goodii, a Recently Described, Rapidly Growing Mycobacterium

    OpenAIRE

    Friedman, N Deborah; Sexton, Daniel J.

    2001-01-01

    We report a case of olecranon bursitis due to Mycobacterium goodii in a 60-year-old man. Prior to recognition of his infection, he received intrabursal steroids and underwent olecranon bursectomy. His infection was cured with antimicrobial therapy consisting of doxycycline and ciprofloxacin. This case illustrates that previously unrecognized members of the Mycobacterium smegmatis group of mycobacteria have pathogenic potential.

  4. Isolamento de Mycobacterium bovis em cão Mycobacterium bovis isolation in a dog

    Directory of Open Access Journals (Sweden)

    P.M.P.C. Mota

    2001-08-01

    Full Text Available This report describes the isolation of Mycobacterium bovis from a dog with a history of co-habitation with bufallos infected with Mycobacterium bovis. After necropsy, the microrganism was isolated from a mesenteric lymphatic node in Stonebrink media and bacterial identification was confirmed by biochemical tests.

  5. Complete Genome Sequences of Field Isolates of Mycobacterium bovis and Mycobacterium caprae.

    Science.gov (United States)

    de la Fuente, José; Díez-Delgado, Iratxe; Contreras, Marinela; Vicente, Joaquín; Cabezas-Cruz, Alejandro; Manrique, Marina; Tobes, Raquel; López, Vladimir; Romero, Beatriz; Domínguez, Lucas; Garrido, Joseba M; Juste, Ramón; Gortazar, Christian

    2015-06-25

    Here we report the complete genome sequences of field isolates of Mycobacterium bovis and the related mycobacterial species, Mycobacterium caprae. The genomes of three M. bovis (MB1, MB3, MB4) and one M. caprae (MB2) field isolates with different virulence, prevalence, and host distribution phenotypes were sequenced.

  6. Copper Homeostasis in Mycobacterium tuberculosis

    Science.gov (United States)

    Shi, Xiaoshan; Darwin, K. Heran

    2015-01-01

    Copper (Cu) is a trace element essential for the growth and development of almost all organisms, including bacteria. However, Cu overload in most systems is toxic. Studies show Cu accumulates in macrophage phagosomes infected with bacteria, suggesting Cu provides an innate immune mechanism to combat invading pathogens. To counteract the host-supplied Cu, increasing evidence suggests that bacteria have evolved Cu resistance mechanisms to facilitate their pathogenesis. In particular, Mycobacterium tuberculosis (Mtb), the causative agent of tuberculosis, has evolved multiple pathways to respond to Cu. Here, we summarize what is currently known about Cu homeostasis in Mtb and discuss potential sources of Cu encountered by this and other pathogens in a mammalian host. PMID:25614981

  7. Genotyping of clinic of Mycobacterium tuberculosis isolates by multilocus variable number of tandem repeat analysis(MLVA)%结核分枝杆菌临床分离株MLVA法分型分析

    Institute of Scientific and Technical Information of China (English)

    文建强; 田卫花; 刘志广; 吕冰; 同重湘; 万康林; 杨枢敏

    2012-01-01

    目的 利用多位点数目可变串联重复序列技术,初步探索甘肃省结核分枝杆菌的基因型及其分布,为防治结核病提供科学依据.方法 选择15个VNTR位点,设计引物,采用PCR扩增、琼脂糖凝胶电泳检测,并利用BioNumerics 4.5软件进行DNA指纹图谱多态性分析.结果 多位点数目可变串联重复序列(MLVA)检测显示,215株结核分枝杆菌呈现7个基因群127种基因型,分别为a、b、c、d、e、f和g基因群,其中e群74.88%( 161/215)为主要流行型(spoligotyping鉴定为北京家族基因型);有抗结核治疗史患者菌株成簇率高于无抗结核治疗史患者,差异有统计学意义(x2 =3.91,P=0.046).结论 兰州地区结核分枝杆菌基因DNA指纹图谱呈现多态性,存在主要流行株,MLVA有助于为当地政府部门制定具体的结核病防治政策和公共卫生应急方案提供科学依据.%Objective To genotype clinic Mycobacterium tuberculosis isolates with multilocus variable number tandem repeat analysis(MLVA) and to provide scientific basis for prevention and control of tuberculosis. Methods Totally 15 variable number tandem repeats( VNTR) loci were analyzed with PCR and agarose gel electrophoresis. The gene diversity was analyzed with BioNumerics 3. 0 software. Results A total of 127 different allele profiles (including 90 unique patterns) were identified by 15 VNTR loci for all 215 isolates of 7 clusters. For all isolates 74. 88% (161/215) were dominant in Gansu province. Univariate analysis showed that the clustered strains were not significantly associated with the resistance to all of four drugs(rifampin,isonicotinyl hydrazide,ethambutol,and streptomycin) .patient's gender,age,regular on irregular therapy,and new cases or relapses (P>0.05 for all) .while significantly associated with antituberculosis therapy history(P> 0.05). Conclusion The strains of Mycobacterium tuberculosis isolated in Gansu province present definitely gene diversity.

  8. Structural features of lipoarabinomannan from Mycobacterium bovis BCG. Determination of molecular mass by laser desorption mass spectrometry.

    Science.gov (United States)

    Venisse, A; Berjeaud, J M; Chaurand, P; Gilleron, M; Puzo, G

    1993-06-15

    It was recently shown that mycobacterial lipoarabinomannan (LAM) can be classified into two types (Chatterjee, D., Lowell, K., Rivoire B., McNeil M. R., and Brennan, P. J. (1992) J. Biol. Chem. 267, 6234-6239) according to the presence or absence of mannosyl residues (Manp) located at the nonreducing end of the oligoarabinosyl side chains. These two types of LAM were found in a pathogenic Mycobacterium tuberculosis strain and in an avirulent M. tuberculosis strain, respectively, suggesting that LAM with Manp characterizes virulent and "disease-inducing strains." We now report the structure of the LAM from Mycobacterium bovis Bacille Calmette-Guérin (BCG) strain Pasteur, largely used throughout the world as vaccine against tuberculosis. Using an up-to-date analytical approach, we found that the LAM of M. bovis BCG belongs to the class of LAMs capped with Manp. By means of two-dimensional homonuclear and heteronuclear scalar coupling NMR analysis and methylation data, the sugar spin system assignments were partially established, revealing that the LAM contained two types of terminal Manp and 2-O-linked Manp. From the following four-step process: (i) partial hydrolysis of deacylated LAM (dLAM), (ii) oligosaccharide derivatization with aminobenzoic ethyl ester, (iii) HPLC purification, (iv) FAB/MS-MS analysis; it was shown that the dimannosyl unit alpha-D-Manp-(1-->2)-alpha-D-Manp is the major residue capping the termini of the arabinan of the LAM. In this report, LAM molecular mass determination was established using matrix-assisted UV-laser desorption/ionization mass spectrometry which reveals that the LAM molecular mass is around 17.4 kDa. The similarity of the LAM structures between M. bovis BCG and M. tuberculosis H37Rv is discussed in regard to their function in the immunopathology of mycobacterial infection.

  9. Computer-assisted prediction of HLA-DR binding and experimental analysis for human promiscuous Th1-cell peptides in the 24 kDa secreted lipoprotein (LppX) of Mycobacterium tuberculosis.

    Science.gov (United States)

    Al-Attiyah, R; Mustafa, A S

    2004-01-01

    The secreted 24 kDa lipoprotein (LppX) is an antigen that is specific for Mycobacterium tuberculosis complex and M. leprae. The present study was carried out to identify the promiscuous T helper 1 (Th1)-cell epitopes of the M. tuberculosis LppX (MT24, Rv2945c) antigen by using 15 overlapping synthetic peptides (25 mers overlapping by 10 residues) covering the sequence of the complete protein. The analysis of Rv2945c sequence for binding to 51 alleles of nine serologically defined HLA-DR molecules, by using a virtual matrix-based prediction program (propred), showed that eight of the 15 peptides of Rv2945c were predicted to bind promiscuously to >/=10 alleles from more than or equal to three serologically defined HLA-DR molecules. The Th1-cell reactivity of all the peptides was assessed in antigen-induced proliferation and interferon-gamma (IFN-gamma)-secretion assays with peripheral blood mononuclear cells (PBMCs) from 37 bacille Calmette-Guérin (BCG)-vaccinated healthy subjects. The results showed that 17 of the 37 donors, which represented an HLA-DR-heterogeneous group, responded to one or more peptides of Rv2945c in the Th1-cell assays. Although each peptide stimulated PBMCs from one or more donors in the above assays, the best positive responses (12/17 (71%) responders) were observed with the peptide p14 (aa 196-220). This suggested a highly promiscuous presentation of p14 to Th1 cells. In addition, the sequence of p14 is completely identical among the LppX of M. tuberculosis, M. bovis and M. leprae, which further supports the usefulness of Rv2945c and p14 in the subunit vaccine design against both tuberculosis and leprosy.

  10. Novel functions of (p)ppGpp and Cyclic di-GMP in mycobacterial physiology revealed by phenotype microarray analysis of wild-type and isogenic strains of Mycobacterium smegmatis.

    Science.gov (United States)

    Gupta, Kuldeepkumar Ramnaresh; Kasetty, Sanjay; Chatterji, Dipankar

    2015-04-01

    The bacterial second messengers (p)ppGpp and bis-(3'-5')-cyclic dimeric GMP (c-di-GMP) regulate important functions, such as transcription, virulence, biofilm formation, and quorum sensing. In mycobacteria, they regulate long-term survival during starvation, pathogenicity, and dormancy. Recently, a Pseudomonas aeruginosa strain lacking (p)ppGpp was shown to be sensitive to multiple classes of antibiotics and defective in biofilm formation. We were interested to find out whether Mycobacterium smegmatis strains lacking the gene for either (p)ppGpp synthesis (ΔrelMsm) or c-di-GMP synthesis (ΔdcpA) would display similar phenotypes. We used phenotype microarray technology to compare the growth of the wild-type and the knockout strains in the presence of several antibiotics. Surprisingly, the ΔrelMsm and ΔdcpA strains showed enhanced survival in the presence of many antibiotics, but they were defective in biofilm formation. These strains also displayed altered surface properties, like impaired sliding motility, rough colony morphology, and increased aggregation in liquid cultures. Biofilm formation and surface properties are associated with the presence of glycopeptidolipids (GPLs) in the cell walls of M. smegmatis. Thin-layer chromatography analysis of various cell wall fractions revealed that the levels of GPLs and polar lipids were reduced in the knockout strains. As a result, the cell walls of the knockout strains were significantly more hydrophobic than those of the wild type and the complemented strains. We hypothesize that reduced levels of GPLs and polar lipids may contribute to the antibiotic resistance shown by the knockout strains. Altogether, our data suggest that (p)ppGpp and c-di-GMP may be involved in the metabolism of glycopeptidolipids and polar lipids in M. smegmatis.

  11. Establishment of Exposure to Organophosphorus Warfare Agents by Means of SPME-GSMS Analysis of Bodily Fluids

    International Nuclear Information System (INIS)

    Reliable chemical analytical procedures for revealing an exposure to toxic chemicals, identifying the active substance, and assessing the degree of exposure are necessary as a component of medical and forensic activities in cases of the possible use of highly toxic chemicals in war conflicts and terrorism acts, as well as emergency situations in chemical industry, specifically at chemical weapons storage and destruction facilities. According to Chemical Weapons Convention, Part XI, Appendix 4, e-17, 'samples of importance in the investigation of alleged use include biomedical samples from human or animal sources (blood, urine, excreta, tissue etc.)'. Urinary metabolites, O-alkyl esters of methylphosphic acid, offer one of the simplest means of confirming an exposure to organophosphorus warfare agents (OPWA). Urine, unlike blood or tissues, does not require invasive collection demanding in terms of sterility. Excretion with urine is the major route of elimination of OPWA from an organism. According to published data, 90% of OPWA metabolites are excreted within 48-72 h after intoxication. We developed an SPME-GCMS procedure for the determination of O-alkyl esters methylphosphonic acid in urine, with the following detection limits,: isopropyl and isobutyl esters 5 ng/ml and pinacolyl ester 1 ng/ml. The procedure involves derivatization of the target compounds directly on the microfiber. The total analysis time is 1-1.5 h. In animal experiments in vivo we could establish the exposure to OPWA at a half-LD50 level within no less than 48 h after intoxication. In principle, OPWA metabolites could be detected in urine within two weeks after intoxication but at higher doses. Retrospective analysis of urinary metabolites in cases of the exposure to low doses of OPWA requires lower detection limits (0.1-1 ng/ml). Optimal objects for the retrospective analysis of OPWA in an organism are long-lived blood protein adducts. We developed a procedure for revealing an exposure to

  12. The Use of the Position Analysis Questionnaire (PAQ) for Establishing the Job Component Validity of Tests. Report No. 5. Final Report.

    Science.gov (United States)

    McCormick, Ernest J.; And Others

    The Position Analysis Questionnaire (PAQ), a structured job analysis questionnaire that provides for the analysis of individual jobs in terms of each of 187 job elements, was used to establish the job component validity of certain commercially-available vocational aptitude tests. Prior to the general analyses reported here, a statistical analysis…

  13. 奶牛分枝杆菌的分离鉴定与耐药性分析%Isolation and identification of Mycobacterium from cows and analysis of their drug resistance features

    Institute of Scientific and Technical Information of China (English)

    邴睿; 田莉莉; 曾巧英; 史兆国; 范伟兴

    2013-01-01

    为优化分枝杆菌PCR诊断方法并监测牛分枝杆菌耐药性,本研究采用罗氏培养基自奶牛淋巴结和肺中分离分枝杆菌,优化本课题组建立的两级多重PCR方法用于分离菌株的分子诊断.一级PCR为两重PCR,靶标16 S rRNA进行属和群的鉴定;二级PCR为七重PCR,根据结核分枝杆菌复合群(Mycobacte-rium tuberculosis complex,MTBC)成员染色体缺失区域的差异,设计了7对引物,依据群内不同种各自呈现特定的扩增谱型进行种的鉴定.对MTBC分离株,全菌水平用绝对浓度法对4种抗结核一线药物(RFP、INH、EMB、SM)进行药敏试验;分子水平用PCR扩增耐药基因rpoB、katG、inhA、embB、rrs,并测序分析.结果显示,本试验共分离出14株分枝杆菌,一级PCR鉴定均为分枝杆菌属成员,其中8株为MTBC成员.二级PCR中,这8株均呈现相同的扩增谱型,均为牛分枝杆菌谱型.药敏试验和耐药基因检测结果一致,8株牛分枝杆菌分离株对4种抗结核病一线药物全部敏感,基因序列分析未见突变.结果表明,本研究优化的两级多重PCR方法具有快速可靠和低成本的优势,是目前国内最新的分子诊断方法.%To optimize the rapid PCR detection method we established previously for Mycobacteria and surveyed the drug resistance of Mycobacteria,in present study,modified Lowenstein-Jensen solid medium was used to isolate Mycobacteria from the cow's lymph node and lung samples.Two level-multiplex PCR were optimized and used to identify isolates.The first level PCR was a duplex PCR targeting special regions corresponding to the genus Mycobacterium and Mycobacterium tuberculosis complex (MTBC) on 16 S rRNA,respectively.The second level PCR was a hepta-plex PCR with 7 pairs of primers targeting different genomic deletion regions of MTBC members,and species were differentiated by amplified profiles particularly corresponding to each species.Drug resistance features for 4 first-line anti

  14. Complete Genome Sequence of Mycobacterium phlei Type Strain RIVM601174

    KAUST Repository

    Abdallah, A. M.

    2012-05-24

    Mycobacterium phlei is a rapidly growing nontuberculous Mycobacterium species that is typically nonpathogenic, with few reported cases of human disease. Here we report the whole genome sequence of M. phlei type strain RIVM601174.

  15. Leg mechanics contribute to establishing swing phase trajectories during memory-guided stepping movements in walking cats: a computational analysis

    Directory of Open Access Journals (Sweden)

    Keir Gordon Pearson

    2015-09-01

    Full Text Available When quadrupeds stop walking after stepping over a barrier with their forelegs, the memory of barrier height and location is retained for many minutes. This memory is subsequently used to guide hind leg movements over the barrier when walking is resumed. The upslope of the initial trajectory of hind leg paw movements is strongly dependent on the initial location of the paw relative to the barrier. In this study, we have attempted to determine whether mechanical factors contribute significantly in establishing the slope of the paw trajectories by creating a 4-link biomechanical model of a cat hind leg and driving this model with a variety of joint-torque profiles, including average torques for a range on initial paw positions relative to the barrier. Torque profiles for individual steps were determined by an inverse dynamic analysis of leg movements in three normal cats. Our study demonstrates that limb mechanics can contribute to establishing the dependency of trajectory slope on the initial position of the paw relative to the barrier. However, an additional contribution of neuronal motor commands was indicated by the fact that the simulated slopes of paw trajectories were significantly less that the observed slopes. A neuronal contribution to the modification of paw trajectories was also revealed by our observations that both the magnitudes of knee flexor muscle EMG bursts and the initial knee flexion torques depended on initial paw position. Previous studies have shown that a shift in paw position prior to stepping over a barrier changes the paw trajectory to be appropriate for the new paw position. Our data indicate that both mechanical and neuronal factors contribute to this updating process, and that any shift in leg position during the delay period modifies the working memory of barrier location.

  16. Genotyping of multi-drug resistant Mycobacterium tuberculosis clinical isolates from Fujian province with multiple loci variable number tandem repeat analysis%福建省耐多药结核分枝杆菌MLVA分型分析

    Institute of Scientific and Technical Information of China (English)

    陈求扬; 赵雁林; 梁庆福; 林建; 林淑芳; 赵永; 魏淑贞; 逄宇; 郑金凤; 王玉锋

    2013-01-01

    目的 了解福建省耐多药结核分枝杆菌分子流行病学特征,为控制耐多药肺结核提供参考依据.方法 采用多位点数目可变串联重复序列基因分型(MLVA)方法,对30个监测点纳入监测的所有耐多药结核分枝杆菌分离菌株DNA进行检测,使用BioNumerics(Version 4.5)软件进行聚类分析.结果 76株耐多药结核分枝杆菌被分为Ⅰ、Ⅱ、Ⅲ三大基因群,分别包含Ⅰ群5株(6.6%)、Ⅱ群68株(89.5%)、Ⅲ群3株(3.9%);在株水平基因分型上,有19株菌成7簇,各包含2~4株菌,成簇菌株来源于同一县区或不同县区.结论 福建省耐多药结核分枝杆菌菌株主要流行株为Ⅱ群菌株;部分菌株存在县区内,甚至跨县域的近期传播流行.%Objective To explore the characteristics of molecular epidemiology of multi-drug Mycobacterium tuberculosis clinical isolates in Fujian province,and to provide reference for multi-drug-resistant tuberculosis(MDR-TB) control. Methods All MDR-TB isolates were selected from 30 survey sites,and the bacterial DNA of these strains were detected by PCR to amplify the loci simultaneously with multiple variable number tandem repeat analysis (MLVA) ,and the clustering of genotypes was analyzed with BioNumerics (Version 4.5). Results Through MLVA, 76 strains of multi- drug resistant Mycobacterium tuberculosis were divided into 3 genogroups,i. e.Ⅰ, Ⅱ , Ⅲ genogroup. The Ⅰ genogroup consists of 5 strains (6. 6% ), Ⅱ genogroup consists of 68 strains ( 89. 5% ), and Ⅲ genogroup consists of 3 strains (3. 9% ). At the strain level, 19 isolates were categorized into 7 clusters,each cluster including 2-4 strains,and the clustering strains were collected from the same county or different county. Conclusion The strain of Ⅱ genogroup is the main epidemic strain in Fujian province, and some genotypes of strains spread recently in some county, even spread across counties. We should strengthen the control of drug resistant

  17. Rv2031c of Mycobacterium tuberculosis: a master regulator of Rv2028-Rv2031 (HspX operon

    Directory of Open Access Journals (Sweden)

    Khurram eMushtaq

    2015-04-01

    Full Text Available AbstractGenes belonging to the same operon are transcribed as a single mRNA molecule in all prokaryotes. The genes of the same operon are presumed to be involved in similar metabolic and physiological processes. Hence, computational analysis of constituent proteins could provide important clues to the functional relationships within the operonic genes. This tends to be more fruitful in the case of Mycobacterium tuberculosis (Mtb, considering the number of hypothetical genes with unknown functions and interacting partners. Dramatic advances in the past decade have increased our knowledge of the mechanisms that tubercle bacilli employ to survive within the host. But the phenomenon of Mtb latency continues to baffle all. Rv2031c belonging to dormancy regulon of Mtb is predominantly expressed during latency, with myriad immunological roles. Thus we attempted to analyze the operon comprising Rv2031c protein to gain insights into its role during latency. In the current study, we have carried out computational analysis of proteins encoded by genes known to be a part of this operon. Our study includes phylogenetic analysis, modeling of protein 3D structures, and protein interaction network analysis. We describe the mechanistic role in the establishment of latency and regulation of DevS/DevR component system. Additionally, we have identified the probable role of these proteins in carbohydrate metabolism, erythromycin tolerance and nucleotide synthesis. Hence, these proteins can modulate the metabolism of mycobacterium inside the host cells and can be important for its survival in latency. The functional characterization and interactome of this important operon can give insight into its role during latency along with the exploitation of constituent proteins as drug targets and vaccine candidates.

  18. Genotyping of Mycobacterium tuberculosis strain isolated from Sichuan province by variable number tandem repeat analysis method%四川省结核分枝杆菌VNTR分型研究

    Institute of Scientific and Technical Information of China (English)

    董海燕; 杨筠; 赵秀芹; 李定越; 陈建; 刘志广; 龙波; 万康林

    2012-01-01

    among Beijing genotype strains. The capability of the VNTR analysis method to differentiate the Mycobacterium tuberculosis strains was improved with increasing VNTR locus. The HGI of 10-locus set was similar to that of 15-locus set. In addition, VNTR showed variation in the ability to differentiate Beijing genotype strains from other geographical areas. Conclusions The discriminatory power of different locus was different, and the same VNTR locus variation in the ability to differentiate Beijing gentry strains from different geographical areas. The 10-locus set was shown to be ideal for use in first-time molecular typing in Sichuan province. The analysis of VNTR data in this study might be useful to select appropriate VNTR loci for the gene-typing of M. tuberculosis in China.

  19. Culture of mycobacterium tuberculosis and analysis of factors related to drug resistance in acounty of lu'an%六安市某县结核病细菌学检测及耐药相关因素分析

    Institute of Scientific and Technical Information of China (English)

    汪全治; 马功燕; 金德兵; 张家武; 孟令宏

    2015-01-01

    Objective To explore the situation and related factors of drug resistance in tuberculosis(TB)patients in lu'an,and to provide evidence for the strategy developing of prevention and control of TB. Methods A county of lu'an was selected by random sampling, and all registered pulmonary TB patients from Jan 2013 to Dec 2013 were enrolled in the study,receiving such laboratory examinations as sputum smear,bacterial culture,drug sensitive test and strain identification. Univariate and multivariate analysis were performed on the relat-ed factors of drug resistance. Results A total of 148 TB patients infected with mycobacterium TB were included,and the total drug resistant rate,extensive resistant rate and multi-drug resistant rate of mycobacterium TB were 25%(37 / 148),4. 58%(3 / 148),8. 1%(12 / 148), respectively. On the other hand,the total drug resistant rate,extensive resistant rate and multi-drug resistant rate of non-mycobacterium TB were 100%(5 / 5),80%(4 / 5)and 80%(4 / 5). Univariate and multivariate analysis showed that,receiving more than once TB treatment was risk factors of mono-resistant and multi drug resistant TB,and 20-50 years old was likely to be a independent risk factor of multi drug re-sistant TB. The smear positive rate was 16. 27%(95 / 584),and the culture positive rate was 26. 20%(153 / 584). Conclusion The high prevalence of drug resistance has been a major challenge for TB control in luˊan. To strengthen the control of untreated and young patients,as to increase the treatment compliance and treatment success rate,reduce the occurrence of drug resistance to TB,is critically important. Meanwhile,to intensify the bacteriology detection of TB is necessary,thereby increasing detective rate of drug resistance and culture positive pulmonary TB.%目的:了解六安市某县结核病耐药状况及相关因素,为制订全市耐药结核病控制策略提供参考依据。方法随机抽取我市辖区1县(区),将2013年1月至12月所有登

  20. Pathogenesis, Immunology, and Diagnosis of Latent Mycobacterium tuberculosis Infection

    Directory of Open Access Journals (Sweden)

    Suhail Ahmad

    2011-01-01

    Full Text Available Phagocytosis of tubercle bacilli by antigen-presenting cells in human lung alveoli initiates a complex infection process by Mycobacterium tuberculosis and a potentially protective immune response by the host. M. tuberculosis has devoted a large part of its genome towards functions that allow it to successfully establish latent or progressive infection in the majority of infected individuals. The failure of immune-mediated clearance is due to multiple strategies adopted by M. tuberculosis that blunt the microbicidal mechanisms of infected immune cells and formation of distinct granulomatous lesions that differ in their ability to support or suppress the persistence of viable M. tuberculosis. In this paper, current understanding of various immune processes that lead to the establishment of latent M. tuberculosis infection, bacterial spreading, persistence, reactivation, and waning or elimination of latent infection as well as new diagnostic approaches being used for identification of latently infected individuals for possible control of tuberculosis epidemic are described.

  1. Rapid and accurate identification of Mycobacterium tuberculosis complex and common non-tuberculous mycobacteria by multiplex real-time PCR targeting different housekeeping genes.

    Science.gov (United States)

    Nasr Esfahani, Bahram; Rezaei Yazdi, Hadi; Moghim, Sharareh; Ghasemian Safaei, Hajieh; Zarkesh Esfahani, Hamid

    2012-11-01

    Rapid and accurate identification of mycobacteria isolates from primary culture is important due to timely and appropriate antibiotic therapy. Conventional methods for identification of Mycobacterium species based on biochemical tests needs several weeks and may remain inconclusive. In this study, a novel multiplex real-time PCR was developed for rapid identification of Mycobacterium genus, Mycobacterium tuberculosis complex (MTC) and the most common non-tuberculosis mycobacteria species including M. abscessus, M. fortuitum, M. avium complex, M. kansasii, and the M. gordonae in three reaction tubes but under same PCR condition. Genetic targets for primer designing included the 16S rDNA gene, the dnaJ gene, the gyrB gene and internal transcribed spacer (ITS). Multiplex real-time PCR was setup with reference Mycobacterium strains and was subsequently tested with 66 clinical isolates. Results of multiplex real-time PCR were analyzed with melting curves and melting temperature (T (m)) of Mycobacterium genus, MTC, and each of non-tuberculosis Mycobacterium species were determined. Multiplex real-time PCR results were compared with amplification and sequencing of 16S-23S rDNA ITS for identification of Mycobacterium species. Sensitivity and specificity of designed primers were each 100 % for MTC, M. abscessus, M. fortuitum, M. avium complex, M. kansasii, and M. gordonae. Sensitivity and specificity of designed primer for genus Mycobacterium was 96 and 100 %, respectively. According to the obtained results, we conclude that this multiplex real-time PCR with melting curve analysis and these novel primers can be used for rapid and accurate identification of genus Mycobacterium, MTC, and the most common non-tuberculosis Mycobacterium species.

  2. Genomes of Two Clinical Isolates of Mycobacterium tuberculosis from Odisha, India

    Science.gov (United States)

    Majid, Mohammad; Qureshi, Asifa; Yerra, Priyadarshini; Kumar, Ashutosh; Kumar, Mandala Kiran; Tiruvayipati, Suma; Baddam, Ramani; Shaik, Sabiha; Srikantam, Aparna

    2014-01-01

    We report whole-genome sequences of two clinical isolates of Mycobacterium tuberculosis isolated from patients in Odisha, India. The sequence analysis revealed that these isolates are of an ancestral type and might represent some of the “pristine” isolates in India that have not admixed with other lineages. PMID:24652981

  3. Differential T-cell recognition of native and recombinant Mycobacterium tuberculosis GroES

    DEFF Research Database (Denmark)

    Rosenkrands, I; Weldingh, K; Ravn, P;

    1999-01-01

    Mycobacterium tuberculosis GroES was purified from culture filtrate, and its identity was confirmed by immunoblot analysis and N-terminal sequencing. Comparing the immunological recognition of native and recombinant GroES, we found that whereas native GroES elicited a strong proliferative response...

  4. Establishment of a maintenance plan based on quantitative analysis in the context of RCM in a JIT production scenario

    International Nuclear Information System (INIS)

    This paper presents a quantitative method for supporting the preparation or review of an equipment maintenance plan in a Just-in-time production scenario. The proposed method includes the following steps: (i) identifying the parts that influence reliability; (ii) surveying the failure rates and times to repair the parts; (iii) classification of parts according to the effect of their failures; (iv) surveying the line occupation parameters; (v) identifying the probability distributions for time to failure, time to repair, and line occupation; (vi) simulating the production and maintenance using the Monte Carlo approach; (vii) conducting a sensitivity analysis concerning variations in demand, MTTF, and MTTR; and (viii) establishing optimized intervals for preventive maintenance. The method is illustrated through an application in a labeling and filling gallons line at a paints and dyes production company. This method allowed the identification of critical parts as it relates to the productive scenario in question. The results can support companies in their decision making regarding the need and/or type of maintenance investment that would best fit an expected demand scenario

  5. Establishment and in-house validation of stem-loop RT PCR method for MicroRNA398 expression analysis

    Directory of Open Access Journals (Sweden)

    Timotijević Gordana S.

    2015-01-01

    Full Text Available MicroRNAs (miRNAs belong to the class of small non-coding RNAs which have important roles throughout development as well as in plant response to diverse environmental stresses. Some of plant miRNAs are essential for regulation and maintenance of nutritive homeostasis when nutrients are in excess or shortage comparing to optimal concentration for certain plant species. Better understanding of miRNAs functions implies development of efficient technology for profiling their gene expression. We set out to establish validate the methodology for miRNA gene expression analysis in cucumber grown under suboptimal mineral nutrient regimes, including iron deficiency. Reverse transcription by “stem-loop” primers in combination with Real time PCR method is one of potential approaches for quantification of miRNA gene expression. In this paper we presented a method for “stem loop” primer design specific for miR398, as well as reaction optimization and determination of Real time PCR efficiency. Proving the accuracy of this method was imperative as “stem loop” RT which consider separate transcription of target and endogenous control. The method was verified by comparison of the obtained data with results of miR398 expression achieved using a commercial kit based on simultaneous conversion of all RNAs in cDNAs. [Projekat Ministarstva nauke Republike Srbije, br. 173005 i br. ON-173028

  6. Combination of multiplex PCR with denaturing high-performance liquid chromatography for rapid detection of Mycobacterium genus and simultaneous identification of the Mycobacterium tuberculosis complex.

    Science.gov (United States)

    Chen, Ru; Gao, Xiao-Bo; Liu, Zhi-Hui; Shen, Xiao-Bing; Guo, Ai-Zhen; Duan, Yan-Yu; Liu, Zhi-Ling; Wu, Xiao-Wei; Zhu, Dao-Zhong

    2013-09-01

    A new assay with the combination of multiplex polymerase chain reaction and denaturing high-performance liquid chromatography analysis was developed for simultaneous detection of Mycobacterium genus and identification of the Mycobacterium tuberculosis complex (MTC). Targeting at genus-specific 16S rRNA sequence of Mycobacterium and specific insertion elements IS6110 and IS1081 of MTC, the assay was validated with 84 strains covering 23 mycobacteria species and 30 strains of non-mycobacteria species. No cross reactivity was observed. Clinical application was carried out on 198 specimens (155 human sputum and 43 bovine tissue samples) and compared with culture. The multiplex assay detected all culture-positive (36 in number) and 35.2% (57/162) culture-negative specimens. The molecular assay was fast that could be completed within 1 h on purified DNA, with the limit of detection as 0.8-1.6 pg per reaction on DNA template. This work provided a useful laboratory tool for rapid identification of Mycobacterium and differentiation of MTC and nontuberculous mycobacteria.

  7. Asymmetric growth and division in Mycobacterium spp.: compensatory mechanisms for non-medial septa.

    Science.gov (United States)

    Singh, Bhupender; Nitharwal, Ram Gopal; Ramesh, Malavika; Pettersson, B M Fredrik; Kirsebom, Leif A; Dasgupta, Santanu

    2013-04-01

    Mycobacterium spp., rod-shaped cells belonging to the phylum Actinomycetes, lack the Min- and Noc/Slm systems responsible for preventing the placement of division sites at the poles or over the nucleoids to ensure septal assembly at mid-cell. We show that the position for establishment of the FtsZ-ring in exponentially growing Mycobacterium marinum and Mycobacterium smegmatis cells is nearly random, and that the cells often divide non-medially, producing two unequal but viable daughters. Septal sites and cellular growth disclosed by staining with the membrane-specific dye FM4-64 and fluorescent antibiotic vancomycin (FL-Vanco), respectively, showed that many division sites were off-centre, often over the nucleoids, and that apical cell growth was frequently unequal at the two poles. DNA transfer through the division septum was detected, and translocation activity was supported by the presence of a putative mycobacterial DNA translocase (MSMEG2690) at the majority of the division sites. Time-lapse imaging of single live cells through several generations confirmed both acentric division site placement and unequal polar growth in mycobacteria. Our evidence suggests that post-septal DNA transport and unequal polar growth may compensate for the non-medial division site placement in Mycobacterium spp. PMID:23387305

  8. Expanding the mycobacterial diversity of metalworking fluids (MWFs): evidence showing MWF colonization by Mycobacterium abscessus.

    Science.gov (United States)

    Kapoor, Renuka; Yadav, Jagjit S

    2012-02-01

    Nontuberculous mycobacteria (NTM) have been associated with hypersensitivity pneumonitis in machinists. Only two species of NTM, namely Mycobacterium immunogenum and Mycobacterium chelonae, have been reported thus far to have the ability to colonize contaminated metalworking fluids (MWFs). Here, we report, for the first time, the presence and characterization (phenotypic and genotypic) of a third species, Mycobacterium abscessus, colonizing these harsh alkaline machining fluids. Two Mycobacterium morphotypes, smooth (S) and rough (R), were isolated (two isolates each) from an in-use industrial MWFs. Biocide susceptibility analysis using triclosan as a model yielded the same minimal inhibitory concentration for the two morphotypes. PCR-restriction analysis-based speciation of the morphotypes confirmed their identity as M. abscessus. Genotyping based on partial DNA sequences corresponding to the variable regions of the hsp65 gene and 16S-23S rRNA operon internal transcribed spacer region and randomly amplified polymorphic DNA-PCR analysis showed that both morphotypes belong to a single genotype. In addition, we isolated and confirmed two novel mycobacterial genotypes, one each of M. immunogenum and M. chelonae from additional in-use MWF screening. Taken together, this study expands the known mycobacterial species- and strain-diversity colonizing MWF. Furthermore, the study emphasizes the need for including M. abscessus species in the existing mycobacterial screening of contaminated MWF. PMID:22092754

  9. Antigenic characterization of dimorphic surface protein in Mycobacterium tuberculosis.

    Science.gov (United States)

    Matsuba, Takashi; Siddiqi, Umme Ruman; Hattori, Toshio; Nakajima, Chie; Fujii, Jun; Suzuki, Yasuhiko

    2016-05-01

    The Mycobacterium tuberculosis Rv0679c protein is a surface protein that contributes to host cell invasion. We previously showed that a single nucleotide transition of the Rv0679c gene leads to a single amino acid substitution from asparagine to lysine at codon 142 in the Beijing genotype family. In this study, we examined the immunological effect of this substitution. Several recombinant proteins were expressed in Escherichia coli and Mycobacterium smegmatis and characterized with antisera and two monoclonal antibodies named 5D4-C2 and 8G10-H2. A significant reduction of antibody binding was detected by enzyme-linked immunosorbent assay (ELISA) and western blot analysis in the Lys142-type protein. This reduction of 8G10-H2 binding was more significant, with the disappearance of a signal in the proteins expressed by recombinant mycobacteria in western blot analysis. In addition, epitope mapping analysis of the recombinant proteins showed a linear epitope by 5D4-C2 and a discontinuous epitope by 8G10-H2. The antibody recognizing the conformational epitope detected only mycobacterial Asn142-type recombinant protein. Our results suggest that a single amino acid substitution of Rv0679c has potency for antigenic change in Beijing genotype strains. PMID:27190237

  10. Uncertainty analysis as essential step in the establishment of the dynamic Design Space of primary drying during freeze-drying.

    Science.gov (United States)

    Mortier, Séverine Thérèse F C; Van Bockstal, Pieter-Jan; Corver, Jos; Nopens, Ingmar; Gernaey, Krist V; De Beer, Thomas

    2016-06-01

    Large molecules, such as biopharmaceuticals, are considered the key driver of growth for the pharmaceutical industry. Freeze-drying is the preferred way to stabilise these products when needed. However, it is an expensive, inefficient, time- and energy-consuming process. During freeze-drying, there are only two main process variables to be set, i.e. the shelf temperature and the chamber pressure, however preferably in a dynamic way. This manuscript focuses on the essential use of uncertainty analysis for the determination and experimental verification of the dynamic primary drying Design Space for pharmaceutical freeze-drying. Traditionally, the chamber pressure and shelf temperature are kept constant during primary drying, leading to less optimal process conditions. In this paper it is demonstrated how a mechanistic model of the primary drying step gives the opportunity to determine the optimal dynamic values for both process variables during processing, resulting in a dynamic Design Space with a well-known risk of failure. This allows running the primary drying process step as time efficient as possible, hereby guaranteeing that the temperature at the sublimation front does not exceed the collapse temperature. The Design Space is the multidimensional combination and interaction of input variables and process parameters leading to the expected product specifications with a controlled (i.e., high) probability. Therefore, inclusion of parameter uncertainty is an essential part in the definition of the Design Space, although it is often neglected. To quantitatively assess the inherent uncertainty on the parameters of the mechanistic model, an uncertainty analysis was performed to establish the borders of the dynamic Design Space, i.e. a time-varying shelf temperature and chamber pressure, associated with a specific risk of failure. A risk of failure acceptance level of 0.01%, i.e. a 'zero-failure' situation, results in an increased primary drying process time

  11. Tuberculosis patients co-infected with Mycobacterium bovis and Mycobacterium tuberculosis in an urban area of Brazil

    Directory of Open Access Journals (Sweden)

    Marcio Roberto Silva

    2013-05-01

    Full Text Available In this cross-sectional study, mycobacteria specimens from 189 tuberculosis (TB patients living in an urban area in Brazil were characterised from 2008-2010 using phenotypic and molecular speciation methods (pncA gene and oxyR pseudogene analysis. Of these samples, 174 isolates simultaneously grew on Löwenstein-Jensen (LJ and Stonebrink (SB-containing media and presented phenotypic and molecular profiles of Mycobacterium tuberculosis, whereas 12 had molecular profiles of M. tuberculosis based on the DNA analysis of formalin-fixed paraffin wax-embedded tissue samples (paraffin blocks. One patient produced two sputum isolates, the first of which simultaneously grew on LJ and SB media and presented phenotypic and molecular profiles of M. tuberculosis, and the second of which only grew on SB media and presented phenotypic profiles of Mycobacterium bovis. One patient provided a bronchial lavage isolate, which simultaneously grew on LJ and SB media and presented phenotypic and molecular profiles of M. tuberculosis, but had molecular profiles of M. bovis from paraffin block DNA analysis, and one sample had molecular profiles of M. tuberculosis and M. bovis identified from two distinct paraffin blocks. Moreover, we found a low prevalence (1.6% of M. bovis among these isolates, which suggests that local health service procedures likely underestimate its real frequency and that it deserves more attention from public health officials.

  12. Mycobacterium marinum: a potential immunotherapy for Mycobacterium tuberculosis infection

    Directory of Open Access Journals (Sweden)

    Tian WW

    2013-07-01

    Full Text Available Wei-wei Tian,1 Qian-qiu Wang,1 Wei-da Liu,2 Jian-ping Shen,1 Hong-sheng Wang11Laboratory of Mycobacterial Disease, Institute of Dermatology, Chinese Academy of Medical Sciences and Peking Union Medical College, Jiangsu Key Laboratory of Molecular Biology for Skin Diseases and STIs, Nanjing, Jiangsu, People’s Republic of China; 2Department of Mycology, Institute of Dermatology, Chinese Academy of Medical Sciences and Peking Union Medical College, Jiangsu Key Laboratory of Molecular Biology for Skin Diseases and STIs, Nanjing, Jiangsu, People’s Republic of ChinaPurpose: The aim of the present study was to investigate the immune response induced by Mycobacterium marinum infection in vitro and the potential of M. marinum as an immunotherapy for M. tuberculosis infection.Methods: The potential human immune response to certain bacillus infections was investigated in an immune cell–bacillus coculture system in vitro. As a potential novel immunotherapy, M. marinum was studied and compared with two other bacilli, Bacillus Calmette-Guérin (BCG and live attenuated M. tuberculosis. We examined the changes in both the bacilli and immune cells, especially the time course of the viability of mycobacteria in the coculture system and host immune responses including multinuclear giant cell formation by Wright–Giemsa modified staining, macrophage polarization by cell surface antigen expression, and cytokines/chemokine production by both mRNA expression and protein secretion.Results: The M. marinum stimulated coculture group showed more expression of CD209, CD68, CD80, and CD86 than the BCG and M. tuberculosis (an attenuated strain, H37Ra groups, although the differences were not statistically significant. Moreover, the M. marinum group expressed more interleukin (IL-1B and IL-12p40 on day 3 (IL-1B: P = 0.003 and 0.004, respectively; IL-12p40: P = 0.001 and 0.011, respectively, a higher level of CXCL10 on day 1 (P = 0.006 and 0.026, respectively, and

  13. Bridging East and West : The Establishment of the International Institute for Applied Systems Analysis (IIASA) in the United States Foreign Policy of Bridge Building, 1964-1972.

    OpenAIRE

    Riska-Campbell, Leena

    2011-01-01

    The dissertation examines the foreign policies of the United States through the prism of science and technology. In the focal point of scrutiny is the policy establishing the International Institute for Applied Systems Analysis (IIASA) and the development of the multilateral part of bridge building in American foreign policy during the 1960s and early 1970s. After a long and arduous negotiation process, the institute was finally established by twelve national member organizations from the fol...

  14. Amplicon DNA melting analysis for the simultaneous detection of Brucella spp and Mycobacterium tuberculosis complex. Potential use in rapid differential diagnosis between extrapulmonary tuberculosis and focal complications of brucellosis.

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    Rocio Sanjuan-Jimenez

    Full Text Available Some sites of extrapulmonary tuberculosis and focal complications of brucellosis are very difficult to differentiate clinically, radiologically, and even histopathologically. Conventional microbiological methods for the diagnosis of extrapulmonary tuberculosis and complicated brucellosis not only lack adequate sensitivity, they are also time consuming, which could lead to an unfavourable prognosis. The aim of this work was to develop a multiplex real-time PCR assay based on SYBR Green I to simultaneously detect Brucella spp and Mycobacterium tuberculosis complex and evaluate the efficacy of the technique with different candidate genes. The IS711, bcsp31 and omp2a genes were used for the identification of Brucella spp and the IS6110, senX3-regX3 and cfp31 genes were targeted for the detection of the M. tuberculosis complex. As a result of the different combinations of primers, nine different reactions were evaluated. A test was defined as positive only when the gene combinations were capable of co-amplifying both pathogens in a single reaction tube and showed distinguishable melting temperatures for each microorganism. According to the melting analysis, only three combinations of amplicons (senX3-regX3+bcsp31, senX3-regX3+IS711 and IS6110+IS711 were visible. Detection limits of senX3-regX3+bcsp31 and senX3-regX3+IS711 were of 2 and 3 genome equivalents for M. tuberculosis complex and Brucella while for IS6110+IS711 they were of 200 and 300 genome equivalents, respectively. The three assays correctly identified all the samples, showing negative results for the control patients. The presence of multicopy elements and GC content were the components most influencing the efficiency of the test; this should be taken into account when designing a multiplex-based SYBR Green I assay. In conclusion, multiplex real time PCR assays based on the targets senX3-regX3+bcsp31 and senX3-regX3+IS711 using SYBR Green I are highly sensitive and reproducible. This may

  15. Effect of growth in biofilms upon antibiotic and chlorine susceptibility of Mycobacterium avium and Mycobacterium intracellulare

    OpenAIRE

    Steed, Keesha

    2003-01-01

    ABSTRACT Mycobacterium avium and Mycobacterium intracellulare are environmental opportunistic pathogens whose source for human infection is water and soil. M. avium and M. intracellulare cause pulmonary infections (tuberculosis) in immunocompetent individuals and bacteremia in immunodeficient individuals (e.g. AIDS). One factor likely influencing the lack of success of antibiotic therapy in patients would be their ability to form biofilms. Growth in biofilms might result in antimicrob...

  16. Genomic Analysis of a Mycobacterium Bovis Bacillus Calmette-Guérin Strain Isolated from an Adult Patient with Pulmonary Tuberculosis

    Science.gov (United States)

    Zhu, Yongqiang; Yu, Xia; Cao, Jun; Wang, Rui; Lv, Xinyan; He, Jin; Guo, Aizhen; Huang, Hairong; Zheng, Huajun; Liu, Siguo

    2015-01-01

    For years, bacillus Calmette-Guérin (BCG) has served as the unique vaccine against tuberculosis and has generally been regarded as safe. However, a clinical strain labeled 3281 that was isolated from a TB patient was identified to be BCG. Via the combination of next-generation sequencing (NGS) and comparative genomic analysis, unique 3281 genetic characteristics were revealed. A region containing the dnaA and dnaN genes that is closely related to the initial chromosome replication was found to repeat three times on the BCG Pasteur-specific tandem duplication region DU1. Due to the minimum number of epitopes in BCG strains, 3281 was inferred to have a high possibility for immune evasion. Additionally, variations in the virulence genes and predictions for potential virulence factors were analyzed. Overall, we report a pathogen that has never previously been thought to be pathogenic and initial insights that are focused on the genetic characteristics of virulent BCG. PMID:25876043

  17. Search for Mycobacterium leprae in wild mammals

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    Sílvia Cristina Barboza Pedrini

    2010-02-01

    Full Text Available Leprosy is still a worldwide public health problem. Brazil and India show the highest prevalence rates of the disease. Natural infection of armadillos Dasypus novemcinctus with Mycobacterium leprae has been reported in some regions of the United States. Identification of bacilli is difficult, particularly due to its inability to grow in vitro. The use of molecular tools represents a fast and sensitive alternative method for diagnosis of mycobacteriosis. In the present study, the diagnostic methods used were bacilloscopy, histopathology, microbiology, and PCR using specific primers for M. leprae repetitive sequences. PCR were performed using genomic DNA extracted from 138 samples of liver, spleen, lymph nodes, and skin of 44 D. novemcinctus, Euphractus sexcinctus, Cabassous unicinctus, and C. tatouay armadillos from the Middle Western region of the state of São Paulo and from the experimental station of Embrapa Pantanal, located in Pantanal da Nhecolândia of Mato Grosso do Sul state. Also, the molecular analysis of 19 samples from internal organs of other road killed species of wild animals, such as Nasua nasua (ring-tailed coati, Procyon cancrivoros (hand-skinned, Cerdocyon thous (dog-pity-bush, Cavia aperea (restless cavy, Didelphis albiventris (skunk, Sphigurrus spinosus (hedgehog, and Gallictis vittata (ferret showed PCR negative data. None of the 157 analyzed samples had shown natural mycobacterial infection. Only the armadillo inoculated with material collected from untreated multibacillary leprosy patient presented PCR positive and its genomic sequencing revealed 100% identity with M. leprae. According to these preliminary studies, based on the used methodology, it is possible to conclude that wild mammals seem not to play an important role in the epidemiology of leprosy in the Middle Western region of the São Paulo state and in the Pantanal of Mato Grosso do Sul state.

  18. Controlling strategy of dormant Mycobacterium tuberculosis

    Institute of Scientific and Technical Information of China (English)

    Gan Yiling; Guo Shuliang

    2014-01-01

    Objective This study aimed to review the available literatures on control of latent tuberculosis (TB) infection and propose a new control strategy to shorten the course of TB chemotherapy.Data sources The data used in this review were mainly obtained from articles listed in PubMed.The search terms were "therapy (treatment) of tuberculosis," "therapy (treatment) of latent TB infection," and "vaccine of TB."Study selection Articles regarding treatment and vaccine of TB were selected and reviewed.Results The most crucial reason causing the prolonged course of TB chemotherapy is the dormant state of Mycobacterium tuberculosis (M.tuberculosis).Nevertheless,there are,to date,no effective drugs that can directly kill the dormant cells of M.tuberculosis in clinical therapy.In accordance with the growth cycle of dormant M.tuberculosis in the body,the methods for controlling dormant M.tuberculosis include direct killing with drugs,prevention of dormant M.tuberculosis resuscitation with vaccines,and resuscitating dormant M.tuberculosis with preparations or drugs and then thoroughly killing these resuscitated M.tuberculosis by using anti-TB therapy.Conclusions The comprehensive analysis of the above three methods suggests that the drugs directly killing dormant cells are in clinical trials,TMC207 is the most beneficial for controlling TB.Because the side effect of vaccines is less and their action period is long,prevention of dormant cells resuscitation with vaccines is promising.The last control method makes it probable that when a huge number of active cells of M.tuberculosis have been killed and eradicated after 1-month short chemotherapy,only a strong short-term subsequent chemotherapy can completely kill and eradicate the remaining M.tuberculosis.This control strategy is expected to significantly shorten the course of TB chemotherapy and bring a new change and breakthrough in TB treatment.

  19. Mycobacterium tuberculosis monoarthritis in a child

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    Rosenberg Alan M

    2008-09-01

    Full Text Available Abstract A child with isolated Mycobacterium tuberculosis monoarthritis, with features initially suggesting oligoarthritis subtype of juvenile idiopathic arthritis, is presented. This patient illustrates the need to consider the possibility of tuberculosis as the cause of oligoarthritis in high-risk pediatric populations even in the absence of a tuberculosis contact history and without evidence of overt pulmonary disease.

  20. Histopathologic features of Mycobacterium ulcerans infection

    NARCIS (Netherlands)

    Guarner, J; Bartlett, J; Whitney, EAS; Raghunathan, PL; Stienstra, Y; Asamoa, K; Etuaful, S; Klutse, E; Quarshie, E; van der Werf, TS; van der Graaf, WTA; King, CH; Ashford, DA

    2003-01-01

    Because of the emergence of Buruli ulcer disease, the World Health Organization launched a Global Buruli Ulcer Initiative in 1998. This indolent skin infection is caused by Mycobacterium ulcerans. During a study of risk factors for the disease in Ghana, adequate excisional skin-biopsy specimens were

  1. Peritoneal tuberculosis due to Mycobacterium caprae

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    T. Nebreda

    2016-01-01

    Full Text Available The incidence of tuberculosis in humans due to Mycobacterium caprae is very low and is almost confined to Europe. We report a case of a previously healthy 41-year-old Moroccan with a 6 month history of abdominal pain, weight loss, fatigue and diarrhea. A diagnosis of peritoneal tuberculosis due to M. caprae was made.

  2. Otomastoiditis Caused by Mycobacterium abscessus, the Netherlands

    NARCIS (Netherlands)

    J. van Ingen; F. Looijmans; P. Mirck; R. Dekhuijzen; M. Boeree; D. van Soolingen

    2010-01-01

    To the Editor: Nontuberculous mycobacteria (NTM) are increasingly recognized as human pathogens (1). Otomastoiditis is a rare extrapulmonary NTM disease type first described in 1976; Mycobacterium chelonae-M. abscessus group bacteria, which are rapidly growing NTM, are the most frequent causative ag

  3. Investigating Mycobacterium chelonae-abscessus Complex

    Centers for Disease Control (CDC) Podcasts

    2011-11-17

    Keith Simmon, scientist at Isentio US discusses research that was done while he was at ARUP laboratories, discusses a new classification of Mycobacterium chelonae-abscessus complex.  Created: 11/17/2011 by National Center for Emerging and Zoonotic Infectious Diseases (NCEZID).   Date Released: 11/22/2011.

  4. Disseminated Mycobacterium chimaera Infection After Cardiothoracic Surgery

    Science.gov (United States)

    Tan, Nicholas; Sampath, Rahul; Abu Saleh, Omar M.; Tweet, Marysia S.; Jevremovic, Dragan; Alniemi, Saba; Wengenack, Nancy L.; Sampathkumar, Priya; Badley, Andrew D.

    2016-01-01

    Ten case reports of disseminated Mycobacterium chimaera infections associated with cardiovascular surgery were published from Europe. We report 3 cases of disseminated M chimaera infections with histories of aortic graft and/or valvular surgery within the United States. Two of 3 patients demonstrated ocular involvement, a potentially important clinical finding.

  5. Analysis and Trend Determination of the Evolution of Tourist Accommodation Establishments (Adjusted Data Based Seasonally in the European Union (28 with Analytical Methods

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    Rodica Pripoaie

    2015-05-01

    Full Text Available This work presents the comparative analysis and trend determination of the evolution tourist accommodation establishments in the European Union (28, adjusted data based seasonally, in the period May 2014 - December 2014 used the Analytical Methods. The principal causes of the evolution tourist accommodation establishments were: the general economic evolution of industries and GDP per capita, the relatively low revenue or low development of the infrastructure. Trend determination of the evolution tourist accommodation establishments in the European Union (28 with analytical methods requires least squares method. On the base the results of the absolute deviations between empirical and theoretical values for the linear, curvilinear and modified exponential regression, will choose the best trend equation for the smallest variation. The best trend model for evolution tourist accommodation establishments in EU (28 is modelled using linear regression equation.

  6. Sub-speciation of Mycobacterium tuberculosis complex from tuberculosis patients in Japan.

    Science.gov (United States)

    Ueyama, Masako; Chikamatsu, Kinuyo; Aono, Akio; Murase, Yoshiro; Kuse, Naoyuki; Morimoto, Kozo; Okumura, Masao; Yoshiyama, Takashi; Ogata, Hideo; Yoshimori, Kozo; Kudoh, Shoji; Azuma, Arata; Gemma, Akihiko; Mitarai, Satoshi

    2014-01-01

    Mycobacterium tuberculosis is the major causative agent of tuberculosis in humans. It is well known that Mycobacterium bovis and other species in the M. tuberculosis complex (MTC) can cause respiratory diseases as zoonosis. We analyzed the MTC isolates collected from tuberculosis patients from Japan in 2002 using a multiplex PCR system that detected cfp32, RD9 and RD12. A total of 970 MTC isolates that were representative of the tuberculosis cases throughout Japan, were examined using this method. As a result, 966 (99.6%) M. tuberculosis, two Mycobacterium africanum and two Mycobacterium canettii were identified using a multiplex PCR system, while no M. bovis was detected. Two isolates that lacked RD9 were initially considered to be M. canettii, but further analysis of the hsp65 sequence revealed them to be M. tuberculosis. Also two M. africanum were identified as M. tuberculosis using the -215 narG nucleotide polymorphism. Though PCR-linked methods have been used for a rapid differentiation of MTC and NTM, from our cases we suggest careful interpretation of RD based identification.

  7. Dielectrophoretic characterization of antibiotic-treated Mycobacterium tuberculosis complex cells.

    Science.gov (United States)

    Inoue, Shinnosuke; Lee, Hyun-Boo; Becker, Annie L; Weigel, Kris M; Kim, Jong-Hoon; Lee, Kyong-Hoon; Cangelosi, Gerard A; Chung, Jae-Hyun

    2015-10-01

    Multi-drug resistant tuberculosis (MDR-TB) has become a serious concern for proper treatment of patients. As a phenotypic method, dielectrophoresis can be useful but is yet to be attempted to evaluate Mycobacterium tuberculosis complex cells. This paper investigates the dielectrophoretic behavior of Mycobacterium bovis (Bacillus Calmette-Guérin, BCG) cells that are treated with heat or antibiotics rifampin (RIF) or isoniazid (INH). The experimental parameters are designed on the basis of our sensitivity analysis. The medium conductivity (σ(m)) and the frequency (f) for a crossover frequency (f(xo1)) test are decided to detect the change of σ(m)-f(xo1) in conjunction with the drug mechanism. Statistical modeling is conducted to estimate the distributions of viable and nonviable cells from the discrete measurement of f (xo1). Finally, the parameters of the electrophysiology of BCG cells, C(envelope) and σ(cyto), are extracted through a sampling algorithm. This is the first evaluation of the dielectrophoresis (DEP) approach as a means to assess the effects of antimicrobial drugs on M. tuberculosis complex cells.

  8. Identification, activity and disulfide connectivity of C-di-GMP regulating proteins in Mycobacterium tuberculosis.

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    Kajal Gupta

    Full Text Available C-di-GMP, a bacterial second messenger plays a key role in survival and adaptation of bacteria under different environmental conditions. The level of c-di-GMP is regulated by two opposing activities, namely diguanylate cyclase (DGC and phosphodiesterase (PDE-A exhibited by GGDEF and EAL domain, respectively in the same protein. Previously, we reported a bifunctional GGDEF-EAL domain protein, MSDGC-1 from Mycobacterium smegmatis showing both these activities (Kumar and Chatterji, 2008. In this current report, we have identified and characterized the homologous protein from Mycobacterium tuberculosis (Rv 1354c named as MtbDGC. MtbDGC is also a bifunctional protein, which can synthesize and degrade c-di-GMP in vitro. Further we expressed Mtbdgc in M. smegmatis and it was able to complement the MSDGC-1 knock out strain by restoring the long term survival of M. smegmatis. Another protein Rv 1357c, named as MtbPDE, is an EAL domain protein and degrades c-di-GMP to pGpG in vitro. Rv1354c and 1357c have seven cysteine amino acids in their sequence, distributed along the full length of the protein. Disulfide bonds play an important role in stabilizing protein structure and regulating protein function. By proteolytic digestion and mass spectrometric analysis of MtbDGC, connectivity between cysteine pairs Cys94-Cys584, Cys2-Cys479 and Cys429-Cys614 was determined, whereas the third cysteine (Cys406 from N terminal was found to be free in MtbDGC protein, which was further confirmed by alkylation with iodoacetamide labeling. Bioinformatics modeling investigations also supported the pattern of disulfide connectivity obtained by Mass spectrometric analysis. Cys406 was mutated to serine by site directed mutagenesis and the mutant MtbC406S was not found to be active and was not able to synthesize or degrade c-di-GMP. The disulfide connectivity established here would help further in understanding the structure - function relationship in MtbDGC.

  9. Proteins with complex architecture as potential targets for drug design: a case study of Mycobacterium tuberculosis.

    Directory of Open Access Journals (Sweden)

    Bálint Mészáros

    2011-07-01

    Full Text Available Lengthy co-evolution of Homo sapiens and Mycobacterium tuberculosis, the main causative agent of tuberculosis, resulted in a dramatically successful pathogen species that presents considerable challenge for modern medicine. The continuous and ever increasing appearance of multi-drug resistant mycobacteria necessitates the identification of novel drug targets and drugs with new mechanisms of action. However, further insights are needed to establish automated protocols for target selection based on the available complete genome sequences. In the present study, we perform complete proteome level comparisons between M. tuberculosis, mycobacteria, other prokaryotes and available eukaryotes based on protein domains, local sequence similarities and protein disorder. We show that the enrichment of certain domains in the genome can indicate an important function specific to M. tuberculosis. We identified two families, termed pkn and PE/PPE that stand out in this respect. The common property of these two protein families is a complex domain organization that combines species-specific regions, commonly occurring domains and disordered segments. Besides highlighting promising novel drug target candidates in M. tuberculosis, the presented analysis can also be viewed as a general protocol to identify proteins involved in species-specific functions in a given organism. We conclude that target selection protocols should be extended to include proteins with complex domain architectures instead of focusing on sequentially unique and essential proteins only.

  10. Enfermedad por Mycobacterium simiae y "Mycobacterium sherrisii" en la Argentina Disease due to Mycobacterium simiae and "Mycobacterium sherrisii" in Argentina

    Directory of Open Access Journals (Sweden)

    Lucía Barrera

    2010-08-01

    Full Text Available Se presenta información reunida retrospectivamente sobre casos de micobacteriosis originados por Mycobacterium simiae (n = 4 y "M. sherrisii" (n = 6. Los casos ocurrieron entre pacientes con sida (n = 6, historia de silicosis (n = 2 o tuberculosis previa (n = 1. Un caso se perdió luego de diagnosticado y nueve fueron tratados con esquemas terapéuticos basados en claritromicina, etambutol y quinolonas. La respuesta fue muy pobre: cinco pacientes fallecieron (cuatro eran HIV positivos, tres permanecieron crónicos y sólo uno curó. Estas micobacterias originaron 2.1% de los casos de micobacteriosis registrados en un período de ocho años. La distinción de estas micobacterias raras de otras más frecuentes por métodos moleculares rápidos, parece ser clínicamente útil para advertir sobre la dificultad que puede presentar el tratamiento. Sin embargo, la diferenciación genotípica entre M. simiae y "M. sherrisii" parecería no ser clínicamente relevante, dado que no quedaron expuestas características que distingan a los pacientes afectados por los dos microorganismos tan estrechamente relacionados.A revision of mycobacterial disease due to M simiae (n = 4 and "M. sherrisii" (n = 6 identified during an eight-year period is presented. Cases occurred among patients with AIDS (n = 6, previous history of silicosis (n = 2 or tuberculosis (n = 2. One case was lost to follow-up and the remaining nine responded poorly to chemotherapy based on clarithromycin, ethambutol and fluoroquinolones. Five patients died of whom four were HIV-positive, three remained chronic and one was cured. These microorganisms originated 2.1% of mycobacterioses cases detected in an eight-year period. Timely identification of this group of uncommon mycobacteria by molecular methods seems to be clinically relevant in order to warn of difficulties inherent to the treatment. However, the distinction between both closely related microorganisms might not be crucial for case

  11. Summary and statistical analysis of environmental monitoring data in the Oarai Research Establishment, Japan Atomic Energy Research Institute

    Energy Technology Data Exchange (ETDEWEB)

    Tamura, Katsuhiro; Kitano, Kyoshiro [Japan Atomic Energy Research Inst., Tokai, Ibaraki (Japan). Tokai Research Establishment; Sibanuma, Yukio; Takasaki, Koichi; Ohhata, Tsutomu

    1998-03-01

    In the Oarai Research Establishment, Japan Atomic Energy Research Institute (JAERI), the environmental monitoring has been conducted for about 29 years since April 1968. The results are discussed for evaluation of long-term and short-term fluctuation in the radiological conditions in the Oarai area. This report summarises the data of the environmental monitoring in Oarai, and statistical analyses were made of the data collected from 1985 through 1994. (author)

  12. Challenges to the establishment of CCAMLR Marine Protected Areas (MPA): A stakeholder analysis of interests and positions

    OpenAIRE

    Lahl, Rebecca

    2015-01-01

    Marine protected areas (MPAs) are used in spatial management for fisheries and conservation purpose. Since the alarming reports on the status of the world’s oceans, MPAs have been on the international agenda for over a decade as they promise various ecological and socioeconomic benefits. The CCAMLR (Commission for the Conservation of Antarctic Marine Living Resources) is the fisheries management regime in the Southern Ocean that is committed to establishing MPAs. Member states have however re...

  13. Analysis of Ethambutol resistance of Mycobacterium tuberculosis derived from tuberculosis patients in Suzhou city%苏州市结核病乙胺丁醇耐药情况分析

    Institute of Scientific and Technical Information of China (English)

    唐佩军; 沈兴华; 虞忻; 王霞芳; 王雪峰; 吴妹英

    2012-01-01

    目的 了解苏州市地区感染结核分枝杆菌(Mycobacterium tuberculosis,MTB)的结核病患者耐乙胺丁醇情况.方法 对2008年9月~2011年5月期间在苏州大学附属传染病医院就诊的肺结核病患者的临床分离株进行菌型鉴定后,用药敏罗氏培养基进行药物敏感性试验,分析结核病患者对一线抗结核药物乙胺丁醇的耐药情况.结果 苏州市地区近三年期结核病耐乙胺丁醇总体发生率为10.7%,其在耐多药病例中的发生率为56.1%;2010~2011年度耐乙胺丁醇发生率明显高于2008~2009年度(χ2=5.54,P<0.05),并且其在耐多药中的发生率也呈显著性差异(χ2=4.30,P<0.05).结论 苏州大学附属传染病医院收治的结核病乙胺丁醇耐药总体发生率和在耐多药中的发生率呈上升趋势.%Objective To investigate the ethambutol resistance of of Mycobacterium tuberculosis derived from tuberculosis patients diagnosed at the affiliated infectious hospital of soochow university in Suzhou. Methods From September 2008 to May 2011, the type of clinical isolates from the hospital were demonstrated and then the ethambutol resistance of Mycobacterium tuberculosis from the patients was analyzed by drug sensitive test. Results The total ethambutol-resistance rate was 10.7% and the ethambutol-resistance cases occupied a percentage of 56. 1 % in the multi-drug resistant tuberculosis. The total ethambutol-resistance rate and its percentage occupied in the multi-drag resistant cases arising during 2010 ~2011 was respectively 12. 3% and 61. 6% , which was profoundly higher than that occurred in 2008 ~ 2009. Conclusion The total ethambutol-resistance rate of Mycobacterium tuberculosis and its rate occurred in the multi-drag resistant cases from the tuberculosis patients in Suzhou city is high and present increasing.

  14. Mycobacterium pseudoshottsii sp. nov., a slowly growing chromogenic species isolated from Chesapeake Bay striped bass (Morone saxatilis)

    Science.gov (United States)

    Rhodes, M.W.; Kator, H.; McNabb, A.; Deshayes, C.; Reyrat, J.-M.; Brown-Elliott, B. A.; Wallace, R.; Trott, K.A.; Parker, J.M.; Lifland, B.; Osterhout, G.; Kaattari, I.; Reece, K.; Vogelbein, W.; Ottinger, C.A.

    2005-01-01

    A group of slowly growing photochromogenic mycobacteria was isolated from Chesapeake Bay striped bass (Morone saxatilis) during an epizootic of mycobacteriosis. Growth characteristics, acid-fastness and 16S rRNA gene sequencing results were consistent with those of the genus Mycobacterium. Biochemical reactions, growth characteristics and mycolic acid profiles (HPLC) resembled those of Mycobacterium shottsii, a non-pigmented mycobacterium also isolated during the same epizootic. Sequencing of the 16S rRNA genes, the gene encoding the exported repeated protein (erp) and the gene encoding the 65 kDa heat-shock protein (hsp65) and restriction enzyme analysis of the hsp65 gene demonstrated that this group of isolates is unique. Insertion sequences associated with Mycobacterium ulcerans, IS2404 and IS2606, were detected by PCR. These isolates could be differentiated from other slowly growing pigmented mycobacteria by their inability to grow at 37 ??C, production of niacin and urease, absence of nitrate reductase, negative Tween 80 hydrolysis and resistance to isoniazid (1 ??g ml-1), p-nitrobenzoic acid, thiacetazone and thiophene-2-carboxylic hydrazide. On the basis of this polyphasic study, it is proposed that these isolates represent a novel species, Mycobacterium pseudoshottsii sp. nov. The type strain, L15T, has been deposited in the American Type Culture Collection as ATCC BAA-883T and the National Collection of Type Cultures (UK) as NCTC 13318T. ?? 2005 IUMS.

  15. Establishment and Analysis of the Sales Model of Fresh Agriculture Food Based on Business to Business E-commerce Platform

    Directory of Open Access Journals (Sweden)

    Xuehui Jiang

    2015-08-01

    Full Text Available This study introduced the network sales model of agricultural products from the view of business to business e-commerce and analyzed the features, we found the sales model was featured by pattern diversity which was however of low level; the existing e-commerce operation scale of agricultural products is small and the anti-risk capacity is weak; its resource integration capacity is insufficient and thus it can’t make full use of the advantages of e-commerce. Based on this, this study established sales chain of fresh agriculture products based on business to business e-commerce, i.e., to trade on the internet; the products are delivered to consumers by third-party according to the delivery of producing area and wholesale market of sales area and then collected by consumers themselves. Meanwhile, we should establish information center of price demand for online trading market and modern refrigeration base of wholesale market, accelerate the development of specialized rural cooperative or industrial company and assure the long-term operation of e-commerce of agricultural products circulation in the future.

  16. Analysis of risk factors and the establishment of a risk model for peripherally inserted central catheter thrombosis

    Institute of Scientific and Technical Information of China (English)

    Fang Hu; Ruo-Nan Hao; Jie Zhang; Zhi-Cheng Ma

    2016-01-01

    Objective: To investigate the main risk factors of peripherally inserted central catheter (PICC) related upper extremity deep venous thrombosis and establish the risk predictive model of PICC-related upper extremity deep venous thrombosis. Methods: Patients with PICC who were hospitalized between January 2014 and July 2015 were studied retrospectively; they were divided into a thrombosis group (n ¼ 52), with patients who had a venous thrombosis complication after PICC, and a no-thrombosis group (n ¼ 144), with patients without venous thrombosis. To compare between the two groups, significantly different variables were selected to perform multivariate logistic regression to establish the risk-predictive model. Results: The PICC catheter history, catheter tip position, and diameter of blood vessel were the key factors for thrombosis. The logistic regression predictive model was as follows:Y ¼ 3.338 þ 2.040 ? PICC catheter history þ1.964? catheter tip position ?1.572? diameter of vessel. The area under the receiver operating characteristic curve for the model was 0.872, 95%CI (0.817e0.927). The cut-off point was 0.801, the sensitivity of the model was 0.832, and the specificity was 0.745. Conclusions: The PICC catheterization history, catheter tip position, the diameter of blood vessel were the key factors for thrombosis. The logistic regression risk model based on these factors is reliable for predicting PICC-related upper extremity deep venous thrombosis.

  17. Powerful methods to establish chromosomal markers in Lactococcus lactis: an analysis of pyrimidine salvage pathway mutants obtained by positive selections

    DEFF Research Database (Denmark)

    Martinussen, Jan; Hammer, Karin

    1995-01-01

    phosphoribosyltransferase (upp), uridindcytidine kinase (udk), pyrimidine nucleoside phosphorylase (pdp), cytidine/deoxycytidine deaminase (dd), thymidine kinase (tdk) and purine nucleoride phosphorylase (pup). Based on an analysis of the mutants obtained, the pathways by which L. lactis metabolizes uracil...

  18. A STRATEGIC ANALYSIS OF THE FUTURE EXPANSION OPTIONS FOR AN ESTABLISHED GAS COOKING APPLIANCES MANUFACTURER IN VIETNAM

    OpenAIRE

    Tang, Yuen San

    2009-01-01

    This paper speaks to the prevailing business environment presently encountered at Kein Hing Industry Sendirian Berhad (KHI), a leading gas stove manufacturer based in Selangor, Malaysia. KHI manufacturers gas stoves under the brand name ?Zenne?. KHI aims to expand into Vietnam. The scope of this paper covers topics such as company overview, external industry analysis, and internal company analysis using strategic tools such as Michael Porter?s five forces framework. This paper will evaluate e...

  19. Frecuencia de micobacterias ambientales en Chile en el año 2008 Frequency of environmental mycobacterium in Chile. 2008

    Directory of Open Access Journals (Sweden)

    Angélica Scappaticcio B

    2011-09-01

    confirmed by Ziehl Neelsen and identification of mycobacteria species or complex were identified by traditional tests according to Runyon classification and biochemical tests, genetic probes and pattern analysis restriction (PRA. Results: 585 cultures were appropriated for inclusion in the study. In 91.3% (n = 534 of the cases Mycobacterium tuberculosis was isolated while 0.3% was Mycobacterium bovis subspecie BCG (n = 3 and 8.4% (n = 48 corresponded to environmental mycobacterium. Of the latter, Mycobacterium kansasii (2.6%, Mycobacterium avium-intracellulare (1.5% and Mycobacterium chelonae (1.0% were the most commonly isolated. Conclusion: According to the figures of this study and comparing them with studies ofprevious years (1988 and 1998 it is concluded that the number of environmental mycobacterium isolated has been relatively constant during the last decade, as well as the species, more commonly isolated.

  20. Mycobacterium fortuitum lipoid pneumonia in a dog.

    Science.gov (United States)

    Leissinger, M K; Garber, J B; Fowlkes, N; Grooters, A M; Royal, A B; Gaunt, S D

    2015-03-01

    A 1-year old female spayed German Shepherd dog was evaluated for acute onset of dyspnea. Pyogranulomatous inflammation and green globoid structures were present on aspirates of the affected lung. Impression smears and histopathology confirmed pyogranulomatous pneumonia, with large amounts of lipid corresponding to the green structures noted cytologically, and identified poorly staining bacterial rods within lipid vacuoles. Special stains confirmed the presence of acid-fast bacterial rods, and polymerase chain reaction and DNA sequencing identified the organism as Mycobacterium fortuitum. M. fortuitum pneumonia is well described in humans and has previously been reported in 4 dogs and 1 cat. Lipid was a prominent cytologic and histologic feature, as is often described in humans and in the single feline case report. Additionally, this case highlights the variable cytologic appearance of lipid, as well as Mycobacterium spp, which are classically nonstaining with Wright-Giemsa. PMID:24788402

  1. Factors that Influence Enterprises’ Enthusiasm for Continuously Establishing the Traceability System: An Empirical Analysis of 81 Enterprises in Sichuan

    Institute of Scientific and Technical Information of China (English)

    2012-01-01

    Agro-product enterprises are the mainstay of the traceability system,and are of great importance. We analyze the factors that influence the enthusiasm of agro-product enterprises for continuously establishing traceability system from characteristics of enterprises,environment and performance,using the data of 81 agro-product enterprises in Sichuan province. It shows that the enthusiasm is greatly influenced by enterprises’ characters and performance,such as age,ownership,business pattern of enterprises,market breadth,condition of enterprises’ traceability system,income,acceptance of consumers,sales volume. Finally we put forward some recommendations,such as consummating the relative policies,increasing the publicity and perfecting performance assessment system.

  2. Establishment of a paclitaxel resistant human breast cancer cell strain (MCF-7/Taxol) and intracellular paclitaxel binding protein analysis.

    Science.gov (United States)

    Zuo, K-Q; Zhang, X-P; Zou, J; Li, D; Lv, Z-W

    2010-01-01

    Multidrug resistance of tumours is one of the most important factors that leads to chemotherapy failure. A multidrug-resistant breast cancer cell line, MCF-7/Taxol, was established from the drug-sensitive parent cell line MCF-7. The biological properties of MCF-7/Taxol, including its drug resistance profile and profile of paclitaxel binding proteins, were analysed and compared with the parent cell line. A number of paclitaxel binding proteins were present in MCF-7 cells but absent from MCF-7/Taxol cells, namely heat shock protein 90, actinin and dermcidin precursor. The identification of differential paclitaxel binding proteins between the multidrug-resistant MCF-7/Taxol cell line and the parent drug-sensitive cell line MCF-7 provides insight into possible mechanisms involved in resistance to these chemotherapy drugs.

  3. A Critical Analysis of the new Typology of Acts in the Draft Treaty Establishing a Constitution for Europe

    Directory of Open Access Journals (Sweden)

    Herwig C. H. Hofmann

    2003-09-01

    Full Text Available This article describes and critically analyses the proposed new typology of acts in the draft treaty establishing a constitution for Europe and its implications for the EU legal system. It comments on the categories of act on the three levels of constitutional law, legislation and implementation. It highlights the importance of the correlation between the catalogue of fundamental rights on one hand and the definition of legislation on the other, which will reform the relation between legislative and executive powers in the Union. The article also uncovers several shortcomings in the proposed typology of acts including the problematic relation between delegated regulations and implementing regulations as well as the lack of adjustment of the proposed system of legal acts to the special nature of the EU.

  4. The preliminary analysis of establishing the cost control system of AP1000 for the Haiyang nuclear power project

    International Nuclear Information System (INIS)

    The AP1000 technology has been first applied to Nuclear Power Plant construction in China. Haiyang Project is the second plant which applies the new technology, and it is the key to the success of the project, that how to control the cost. The cost control of AP1000 is to manage and monitor all the cost of the project, including the cost of project management, design, procurement, construction, and startup/commissioning. For the Haiyang Project, the cost control system should be established to ensure that the evaluation of the procurement order should be covered in the original budget, and all potential commitments are evaluated and approved within the confinement of cost control, and reduce the risk of the first reactor and get the most profit. (authors)

  5. Stable isotope analysis of a newly established macrofaunal food web 1.5 years after the Hebei Spirit oil spill.

    Science.gov (United States)

    Han, Eunah; Park, Hyun Je; Bergamino, Leandro; Choi, Kwang-Sik; Choy, Eun Jung; Yu, Ok Hwan; Lee, Tae Won; Park, Heung-Sik; Shim, Won Joon; Kang, Chang-Keun

    2015-01-15

    We examined trophic relationships in a newly established community 1.5 years after the Hebei Spirit oil spill on the west coast of Korea. Carbon and nitrogen stable isotope ratios in consumers and their potential food sources were compared between the oil-spill site and reference site, located 13.5 km from the oil-spill spot. The isotopic mixing model and a novel circular statistics rejected the influx of petrogenic carbon into the community and identified spatial consistencies such as the high contributions of microphytobenthos, food-chain length, and the isotopic niche of each feeding guild between sites. We suggested that high level of trophic plasticity and the prevalence of omnivory of consumers may promote the robustness of food web against the oil contamination. Furthermore, we highlighted the need of holistic approaches including different functional groups to quantify changes in the food web structure and assess the influence of different perturbations including oil spill. PMID:25467873

  6. Mycobacterium fortuitum abdominal wall abscesses following liposuction

    OpenAIRE

    Al Soub, Hussam; Al-Maslamani, Eman; Al-Maslamani, Mona

    2008-01-01

    We describe here a case of abdominal abscesses due to Mycobacterium fortuitum following liposuction. The abscesses developed three months after the procedure and diagnosis was delayed for five months. The clues for diagnosis were persistent pus discharge in spite of broad spectrum antibiotics and failure to grow any organisms on routine culture. This condition has been rarely reported; however, the increasing number of liposuction procedures done and awareness among physicians will probably r...

  7. Autophagy in Mycobacterium tuberculosis and HIV infections

    OpenAIRE

    Espert, Lucile; Beaumelle, Bruno; Vergne, Isabelle

    2015-01-01

    Human Immunodeficiency Virus (HIV) and Mycobacterium tuberculosis (M.tb) are among the most lethal human pathogens worldwide, each being responsible for around 1.5 million deaths annually. Moreover, synergy between acquired immune deficiency syndrome (AIDS) and tuberculosis (TB) has turned HIV/M.tb co-infection into a major public health threat in developing countries. In the past decade, autophagy, a lysosomal catabolic process, has emerged as a major host immune defense mechanism against in...

  8. Mycobacterium kansasii Pulmonary Diseases in Korea

    OpenAIRE

    Yim, Jae-Joon; Park, Young-Kil; Lew, Woo Jin; Bai, Gill-Han; Han, Sung Koo; Shim, Young-Soo

    2005-01-01

    Mycobacterium kansasii is one of the most common cause of pulmonary diseases due to nontuberculous mycobacteria. We investigated the changing in the number of isolation of M. kansasii and the clinical characteristics of M. kansasii pulmonary disease in Korea. Through searching the database of the Korean Institute of Tuberculosis, we identified the cases of isolated M. kansasii from 1992 to 2002. The number of M. kansasii isolation had increased from once in 1992 to 62 in 2002. Fifteen patient...

  9. A Dermal Piercing Complicated by Mycobacterium fortuitum

    Directory of Open Access Journals (Sweden)

    Trisha Patel

    2013-01-01

    Full Text Available Background. Dermal piercings have recently become a fashion symbol. Common complications include hypertrophic scarring, rejection, local infection, contact allergy, and traumatic tearing. We report a rare case of Mycobacterium fortuitum following a dermal piercing and discuss its medical implications and treatments. Case. A previously healthy 19-year-old woman presented complaining of erythema and edema at the site of a dermal piercing on the right fourth dorsal finger. She was treated with a 10-day course of trimethoprim-sulfamethoxazole and one course of cephalexin by her primary care physician with incomplete resolution. The patient stated that she had been swimming at a local water park daily. A punch biopsy around the dermal stud was performed, and cultures with sensitivities revealed Mycobacterium fortuitum. The patient was treated with clarithromycin and ciprofloxacin for two months receiving full resolution. Discussion. Mycobacterium fortuitum is an infrequent human pathogen. This organism is a Runyon group IV, rapidly growing nontuberculous mycobacteria, often found in water,soil, and dust. Treatment options vary due to the size of the lesion. Small lesions are typically excised, while larger lesions require treatment for 2–6 months with antibiotics. We recommend a high level of suspicion for atypical mycobacterial infections in a piercing resistant to other therapies.

  10. Mycobacterium paratuberculosis. Factors that influence mycobactin dependence.

    Science.gov (United States)

    Lambrecht, R S; Collins, M T

    1992-01-01

    Mycobacterium paratuberculosis does not produce any detectable mycobactin, an iron-binding compound that is synthesized by most Mycobacterium spp. and necessary for the growth of all mycobacteria. This study examined the influence of various culture conditions on mycobactin dependence in M. paratuberculosis. Using a radiometric growth assay, we found the minimal concentration of mycobactin J necessary for growth of M. paratuberculosis to be 0.006 microM, whereas 1.2 microM (1 microgram/ml) was required for optimal growth. In media without mycobactin at iron concentrations less than or equal to 100 microM, growth of M. paratuberculosis occurred at pH 5.0, but not pH 6.8. Iron concentrations greater than 100 microM did not significantly increase growth at pH 5.0, but at pH 6.8 the growth rate increased with increasing amounts of iron reaching a rate equal to control cultures containing mycobactin. Mycobacterium paratuberculosis appeared to lose mycobactin dependence when subcultured; however, this was subsequently shown to be a result of mycobactin carried over from primary medium. Removal of this contaminating cell-wall-associated mycobactin reestablished mycobactin dependence. We conclude that mycobactin dependence must be carefully determined because it is a key test used in identification of M. paratuberculosis and may be easily influenced by media pH, iron concentration, and mycobactin carryover from primary media. PMID:1582168

  11. Latin-American-Mediterranean lineage of Mycobacterium tuberculosis: Human traces across pathogen's phylogeography.

    Science.gov (United States)

    Mokrousov, Igor; Vyazovaya, Anna; Iwamoto, Tomotada; Skiba, Yuriy; Pole, Ilva; Zhdanova, Svetlana; Arikawa, Kentaro; Sinkov, Viacheslav; Umpeleva, Tatiana; Valcheva, Violeta; Alvarez Figueroa, Maria; Ranka, Renate; Jansone, Inta; Ogarkov, Oleg; Zhuravlev, Viacheslav; Narvskaya, Olga

    2016-06-01

    Currently, Mycobacterium tuberculosis isolates of Latin-American Mediterranean (LAM) family may be detected far beyond the geographic areas that coined its name 15years ago. Here, we established the framework phylogeny of this geographically intriguing and pathobiologically important mycobacterial lineage and hypothesized how human demographics and migration influenced its phylogeography. Phylogenetic analysis of LAM isolates from all continents based on 24 variable number of tandem repeats (VNTR) loci and other markers identified three global sublineages with certain geographic affinities and defined by large deletions RD115, RD174, and by spoligotype SIT33. One minor sublineage (spoligotype SIT388) appears endemic in Japan. One-locus VNTR signatures were established for sublineages and served for their search in published literature and geographic mapping. We suggest that the LAM family originated in the Western Mediterranean region. The most widespread RD115 sublineage seems the most ancient and encompasses genetically and geographically distant branches, including extremely drug resistant KZN in South Africa and LAM-RUS recently widespread across Northern Eurasia. The RD174 sublineage likely started its active spread in Brazil; its earlier branch is relatively dominated by isolates from South America and the derived one is dominated by Portuguese and South/Southeastern African isolates. The relatively most recent SIT33-sublineage is marked with enigmatic gaps and peaks across the Americas and includes South African clade F11/RD761, which likely emerged within the SIT33 subpopulation after its arrival to Africa. In addition to SIT388-sublineage, other deeply rooted, endemic LAM sublineages may exist that remain to be discovered. As a general conclusion, human mass migration appears to be the major factor that shaped the M. tuberculosis phylogeography over large time-spans. PMID:27001605

  12. Microbial degradation of the multiply branched alkane 2,6,10,15,19, 23-hexamethyltetracosane (Squalane) by Mycobacterium fortuitum and Mycobacterium ratisbonense.

    Science.gov (United States)

    Berekaa, M M; Steinbüchel, A

    2000-10-01

    Among several bacterial species belonging to the general Gordonia, Mycobacterium, Micromonospora, Pseudomonas, and Rhodococcus, only two mycobacterial isolates, Mycobacterium fortuitum strain NF4 and the new isolate Mycobacterium ratisbonense strain SD4, which was isolated from a sewage treatment plant, were capable of utilizing the multiply branched hydrocarbon squalane (2,6,10,15,19, 23-hexamethyltetracosane) and its analogous unsaturated hydrocarbon squalene as the sole carbon source for growth. Detailed degradation studies and high-pressure liquid chromatography analysis showed a clear decrease of the concentrations of squalane and squalene during biomass increase. These results were supported by resting-cell experiments using strain SD4 and squalane or squalene as the substrate. The degradation of acyclic isoprenoids and alkanes as well as of acids derived from these compounds was also investigated. Inhibition of squalane and squalene degradation by acrylic acid indicated the possible involvement of beta-oxidation in the degradation route. To our knowledge, this is the first report demonstrating the biodegradation of squalane by using defined axenic cultures.

  13. A robust SNP barcode for typing Mycobacterium tuberculosis complex strains

    Science.gov (United States)

    Coll, Francesc; McNerney, Ruth; Guerra-Assunção, José Afonso; Glynn, Judith R.; Perdigão, João; Viveiros, Miguel; Portugal, Isabel; Pain, Arnab; Martin, Nigel; Clark, Taane G.

    2014-01-01

    Strain-specific genomic diversity in the Mycobacterium tuberculosis complex (MTBC) is an important factor in pathogenesis that may affect virulence, transmissibility, host response and emergence of drug resistance. Several systems have been proposed to classify MTBC strains into distinct lineages and families. Here, we investigate single-nucleotide polymorphisms (SNPs) as robust (stable) markers of genetic variation for phylogenetic analysis. We identify ~92k SNP across a global collection of 1,601 genomes. The SNP-based phylogeny is consistent with the gold-standard regions of difference (RD) classification system. Of the ~7k strain-specific SNPs identified, 62 markers are proposed to discriminate known circulating strains. This SNP-based barcode is the first to cover all main lineages, and classifies a greater number of sublineages than current alternatives. It may be used to classify clinical isolates to evaluate tools to control the disease, including therapeutics and vaccines whose effectiveness may vary by strain type. PMID:25176035

  14. First isolation ofMycobacterium setense from hospital water

    Institute of Scientific and Technical Information of China (English)

    Davood Azadi; Abass Daei Naser; Hasan Shojaei

    2016-01-01

    Objective:To present the findings of a study on isolation of four unrelated environmental strains ofMycobacterium setense (M. setense) from hospital environment and help to assess the natural habitat and the mode of transmission in man. Methods: The water samples were collected from hospital departments and cultured on Löwenstein-Jensen and Sauton's media. The isolates, i.e.,AW3-2,AW5,AW11 andAW18 were subjected to identification by conventional and molecular tests including sequencing analysis of16S rRNA. Results: The water isolates revealed the phenotypic and molecular features which were consistent withM. setense including a genus specific amplicon of thehsp65 gene and 99.6% similarities with those of M. setenseCIP:109395T16S rRNA gene sequences. Conclusions: The current report will contribute to a better understanding of the pathogenesis and path of transmission of this opportunistic pathogen to human.

  15. Co-infecção por Mycobacterium tuberculosis e vírus da imunodeficiência humana: uma análise epidemiológica em Taubaté (SP Co-infection with Mycobacterium tuberculosis and human immunodeficiency virus: an epidemiological analysis in the city of Taubaté, Brazil

    Directory of Open Access Journals (Sweden)

    Luiz Gustavo Miranda de Carvalho

    2006-10-01

    Full Text Available OBJETIVO: Discutir os principais aspectos da co-infecção por Mycobacterium tuberculosis e vírus da imunodeficiência humana no município de Taubaté (SP nos anos de 2001 e 2002. MÉTODOS: Este trabalho apresenta o levantamento epidemiológico dos casos de tuberculose ocorridos em Taubaté em 2001 e 2002. RESULTADOS: Foram analisados 250 casos de tuberculose, dos quais 70 corresponderam a casos de sorologia positiva para o vírus da imunodeficiência humana (28%, 95 de sorologia negativa (38%, e para 85 pacientes a sorologia não foi realizada (34%. Com relação ao primeiro grupo houve predomínio do sexo masculino e da faixa etária de 30 a 40 anos, a forma clínica de tuberculose mais comum foi a pulmonar (65,71% e a taxa de cura foi de 59,38% . No grupo de pacientes para os quais não se comprovou a co-infecção também predominou o sexo masculino e ocorrência na mesma faixa etária, sendo a forma clínica mais comum também a pulmonar (70,55% e a taxa de cura foi de 81,63%. CONCLUSÃO: Concluiu-se que o vírus da imunodeficiência humana é importante na epidemiologia da tuberculose e, portanto, o teste sorológico para o vírus da imunodeficiência humana deve ser realizado quando se diagnostica a presença de tuberculose.OBJECTIVE: To discuss the main aspects of co-infection with Mycobacterium tuberculosis and human immunodeficiency virus in the city of Taubaté (located in the state of São Paulo, Brazil in 2001 and 2002. METHODS: This study presents epidemiological data on tuberculosis cases occurring in Taubaté in 2001 and 2002. RESULTS: Of the 250 cases of tuberculosis analyzed, 70 (28% presented human immunodeficiency virus seropositivity, 95 (38% presented human immunodeficiency virus seronegativity, and 85 (34% were patients who had not been submitted to serological testing. In the first group (tuberculosis and human immunodeficiency virus seropositivity, males from 30 to 40 years of age predominated, the most common clinical

  16. Dynamic Response of Mycobacterium vanbaalenii PYR-1 to BP Deepwater Horizon Crude Oil

    OpenAIRE

    Kim, Seong-Jae; Kweon, Ohgew; Sutherland, John B.; Kim, Hyun-Lee; Jones, Richard C; Burback, Brian L.; Graves, Steven W.; Psurny, Edward; Cerniglia, Carl E.

    2015-01-01

    We investigated the response of the hydrocarbon-degrading Mycobacterium vanbaalenii PYR-1 to crude oil from the BP Deepwater Horizon (DWH) spill, using substrate depletion, genomic, and proteome analyses. M. vanbaalenii PYR-1 cultures were incubated with BP DWH crude oil, and proteomes and degradation of alkanes and polycyclic aromatic hydrocarbons (PAHs) were analyzed at four time points over 30 days. Gas chromatography-mass spectrometry (GC-MS) analysis showed a chain length-dependent patte...

  17. Construction and application of a co-expression network in Mycobacterium tuberculosis

    OpenAIRE

    Jun Jiang; Xian Sun; Wei Wu; Li Li; Hai Wu; Lu Zhang; Guohua Yu; Yao Li

    2016-01-01

    Because of its high pathogenicity and infectivity, tuberculosis is a serious threat to human health. Some information about the functions of the genes in Mycobacterium tuberculosis genome was currently available, but it was not enough to explore transcriptional regulatory mechanisms. Here, we applied the WGCNA (Weighted Gene Correlation Network Analysis) algorithm to mine pooled microarray datasets for the M. tuberculosis H37Rv strain. We constructed a co-expression network that was subdivide...

  18. Mycobacterium avium subsp. paratuberculosis Strains Isolated from Crohn's Disease Patients and Animal Species Exhibit Similar Polymorphic Locus Patterns

    OpenAIRE

    Ghadiali, Alifiya H.; Strother, Megan; Naser, Saleh A.; Manning, Elizabeth J. B.; Sreevatsan, Srinand

    2004-01-01

    Analysis of short sequence repeats of Mycobacterium avium subsp. paratuberculosis isolated from Crohn's disease patients identified two alleles, both of which clustered with strains derived from animals with Johne's disease. Identification of a limited number of genotypes among human strains implies the existence of human disease-associated genotypes and strain sharing with animals.

  19. Establishment of cDNA Microarray Analysis at the Genomic Medicine Research Core Laboratory (GMRCL) of Chang Gung Memorial Hospital .

    OpenAIRE

    Tzu-Hao Wang; Yun-Shien Lee; En-Shih Chen; Wei-Hsiang Kong; Lung-Kun Chen; Ding-Wei Hsueh; Min-Li Wei; Hsing-Shih Wang; Ying-Shiung Lee

    2004-01-01

    Background: Advances in molecular and computational biology have led to the developmentof powerful, high-throughput methods for analysis of differential geneexpression, which are opening up new opportunities in genomic medicine.DNA microarray technology has been enthusiastically integrated into basicbiomedical research and will eventually become a molecular monitoring toolfor various clinical courses.Methods: As a core research facility of Chang Gung University (CGU) and ChangGung Memorial Ho...

  20. Identifying past fire regimes throughout the Holocene in Ireland using new and established methods of charcoal analysis

    Science.gov (United States)

    Hawthorne, Donna; Mitchell, Fraser J. G.

    2016-04-01

    Globally, in recent years there has been an increase in the scale, intensity and level of destruction caused by wildfires. This can be seen in Ireland where significant changes in vegetation, land use, agriculture and policy, have promoted an increase in fires in the Irish landscape. This study looks at wildfire throughout the Holocene and draws on lacustrine charcoal records from seven study sites spread across Ireland, to reconstruct the past fire regimes recorded at each site. This work utilises new and accepted methods of fire history reconstruction to provide a recommended analytical procedure for statistical charcoal analysis. Digital charcoal counting was used and fire regime reconstructions carried out via the CharAnalysis programme. To verify this record new techniques are employed; an Ensemble-Member strategy to remove the objectivity associated with parameter selection, a Signal to Noise Index to determine if the charcoal record is appropriate for peak detection, and a charcoal peak screening procedure to validate the identified fire events based on bootstrapped samples. This analysis represents the first study of its kind in Ireland, examining the past record of fire on a multi-site and paleoecological timescale, and will provide a baseline level of data which can be built on in the future when the frequency and intensity of fire is predicted to increase.

  1. Transition Metal–α-Amino Acid Complexes with Antibiotic Activity against Mycobacterium spp.

    OpenAIRE

    Karpin, George W.; Merola, Joseph S.; Joseph O. Falkinham

    2013-01-01

    Synthetic iridium-, rhodium-, and ruthenium-amino acid complexes with hydrophobic l-amino acids have antibiotic activity against Mycobacterium spp., including Mycobacterium bovis BCG and the rapidly growing species Mycobacterium abscessus and Mycobacterium chelonae. Concentrations of transition metal-amino acid complexes demonstrating hemolysis or cytotoxicity were 10- to 25-fold higher than were the MICs.

  2. Surviving within the amoebal exocyst: the Mycobacterium avium complex paradigm

    Directory of Open Access Journals (Sweden)

    Drancourt Michel

    2010-04-01

    Full Text Available Abstract Background Most of environmental mycobacteria have been previously demonstrated to resist free-living amoeba with subsequent increased virulence and resistance to antibiotics and biocides. The Mycobacterium avium complex (MAC comprises of environmental organisms that inhabit a wide variety of ecological niches and exhibit a significant degree of genetic variability. We herein studied the intra-ameobal location of all members of the MAC as model organisms for environmental mycobacteria. Results Type strains for M. avium, Mycobacterium intracellulare, Mycobacterium chimaera, Mycobacterium colombiense, Mycobacterium arosiense, Mycobacterium marseillense, Mycobacterium timonense and Mycobacterium bouchedurhonense were co-cultivated with the free-living amoeba Acanthamoeba polyphaga strain Linc-AP1. Microscopic analyses demonstrated the engulfment and replication of mycobacteria into vacuoles of A. polyphaga trophozoites. Mycobacteria were further entrapped within amoebal cysts, and survived encystment as demonstrated by subculturing. Electron microscopy observations show that, three days after entrapment into A. polyphaga cysts, all MAC members typically resided within the exocyst. Conclusions Combined with published data, these observations indicate that mycobacteria are unique among amoeba-resistant bacteria, in residing within the exocyst.

  3. Risk factors for Mycobacterium tuberculosis infection among children in Greenland

    DEFF Research Database (Denmark)

    Søborg, Bolette; Andersen, Aase Bengaard; Melbye, Mads;

    2011-01-01

    To examine the risk factors for Mycobacterium tuberculosis infection (MTI) among Greenlandic children for the purpose of identifying those at highest risk of infection.......To examine the risk factors for Mycobacterium tuberculosis infection (MTI) among Greenlandic children for the purpose of identifying those at highest risk of infection....

  4. Complete Genome Sequence of Mycobacterium vaccae Type Strain ATCC 25954

    KAUST Repository

    Ho, Y. S.

    2012-10-26

    Mycobacterium vaccae is a rapidly growing, nontuberculous Mycobacterium species that is generally not considered a human pathogen and is of major pharmaceutical interest as an immunotherapeutic agent. We report here the annotated genome sequence of the M. vaccae type strain, ATCC 25954.

  5. Establishment of Intestinal Bacteriology

    OpenAIRE

    Mitsuoka, Tomotari

    2014-01-01

    Research on intestinal bacteria began around the end of the 19th century. During the last 5 decades of the 20th century, research on the intestinal microbiota made rapid progress. At first, in my work, I first developed a method of comprehensive analysis of the intestinal microbiota, and then I established classification and identification methods for intestinal anaerobes. Using these methods I discovered a number of ecological rules governing the intestinal microbiota and the role of the int...

  6. A PULMONARY INFECTION CAUSED BY MYCOBACTERIUM PEREGRINUM– A CASE REPORT.

    Directory of Open Access Journals (Sweden)

    Tatina T. Todorova

    2015-12-01

    Full Text Available Mycobacterium peregrinum is a member of the group of rapidly growing Nontuberculous Mycobacteria (NTM. It can be found in high frequency in natural and laboratory environments and is considered to be uncommonrare pathogen for both immunocompetent and immunosuppressed individuals. Currently, pulmonary infections caused by Mycobacterium peregrinum are unusual and diagnosed only in limited number of cases. Here, we present a clinical case of elderly man (72 years with 1 month history of non-specific respiratory symptomatic. The patient was without underlying immunosuppressive condition or lung disease. Chest X-ray demonstrated persistent pleural effusion, opacities and cavitations in the right lobe. One of the sputum culturesgrewa rapidly growing mycobacterium and the isolated strain was found to be Mycobacterium peregrinumas identified by molecular genetic detection (PCR and DNA strip technology. To our knowledge, this is the third case in the world to report Mycobacterium peregrinumas a possible causative agent of pulmonary infection.

  7. The benchmark analysis of gastric, colorectal and rectal cancer pathways: toward establishing standardized clinical pathway in the cancer care.

    Science.gov (United States)

    Ryu, Munemasa; Hamano, Masaaki; Nakagawara, Akira; Shinoda, Masayuki; Shimizu, Hideaki; Miura, Takeshi; Yoshida, Isao; Nemoto, Atsushi; Yoshikawa, Aki

    2011-01-01

    Most clinical pathways in treating cancers in Japan are based on individual physician's personal experiences rather than on an empirical analysis of clinical data such as benchmark comparison with other hospitals. Therefore, these pathways are far from being standardized. By comparing detailed clinical data from five cancer centers, we have observed various differences among hospitals. By conducting benchmark analyses, providing detailed feedback to the participating hospitals and by repeating the benchmark a year later, we strive to develop more standardized clinical pathways for the treatment of cancers. The Cancer Quality Initiative was launched in 2007 by five cancer centers. Using diagnosis procedure combination data, the member hospitals benchmarked their pre-operative and post-operative length of stays, the duration of antibiotics administrations and the post-operative fasting duration for gastric, colon and rectal cancers. The benchmark was conducted by disclosing hospital identities and performed using 2007 and 2008 data. In the 2007 benchmark, substantial differences were shown among five hospitals in the treatment of gastric, colon and rectal cancers. After providing the 2007 results to the participating hospitals and organizing several brainstorming discussions, significant improvements were observed in the 2008 data study. The benchmark analysis of clinical data is extremely useful in promoting more standardized care and, thus in improving the quality of cancer treatment in Japan. By repeating the benchmark analyses, we can offer truly clinical evidence-based higher quality standardized cancer treatment to our patients.

  8. Requirements for Defining Utility Drive Cycles: An Exploratory Analysis of Grid Frequency Regulation Data for Establishing Battery Performance Testing Standards

    Energy Technology Data Exchange (ETDEWEB)

    Hafen, Ryan P.; Vishwanathan, Vilanyur V.; Subbarao, Krishnappa; Kintner-Meyer, Michael CW

    2011-10-19

    Battery testing procedures are important for understanding battery performance, including degradation over the life of the battery. Standards are important to provide clear rules and uniformity to an industry. The work described in this report addresses the need for standard battery testing procedures that reflect real-world applications of energy storage systems to provide regulation services to grid operators. This work was motivated by the need to develop Vehicle-to-Grid (V2G) testing procedures, or V2G drive cycles. Likewise, the stationary energy storage community is equally interested in standardized testing protocols that reflect real-world grid applications for providing regulation services. As the first of several steps toward standardizing battery testing cycles, this work focused on a statistical analysis of frequency regulation signals from the Pennsylvania-New Jersey-Maryland Interconnect with the goal to identify patterns in the regulation signal that would be representative of the entire signal as a typical regulation data set. Results from an extensive time-series analysis are discussed, and the results are explained from both the statistical and the battery-testing perspectives. The results then are interpreted in the context of defining a small set of V2G drive cycles for standardization, offering some recommendations for the next steps toward standardizing testing protocols.

  9. The location of midfacial landmarks according to the method of establishing the midsagittal reference plane in three-dimensional computed tomography analysis of facial asymmetry

    Energy Technology Data Exchange (ETDEWEB)

    Kim, Min Sun; Lee, Eun Joo; Lee, Jae Seo; Kang, Byung Cheock; Yoon, Suk Ja [Dental Science Research Institute, Chonnam National University, Gwangju (Korea, Republic of); Song, In Ja [Dept. of Nursing, Kwangju Women' s University, Gwangju (Korea, Republic of)

    2015-12-15

    The purpose of this study was to evaluate the influence of methods of establishing the midsagittal reference plane (MRP) on the locations of midfacial landmarks in the three-dimensional computed tomography (CT) analysis of facial asymmetry. A total of 24 patients (12 male and 12 female; mean age, 22.5 years; age range, 18.2-29.7 years) with facial asymmetry were included in this study. The MRP was established using two different methods on each patient's CT image. The x-coordinates of four midfacial landmarks (the menton, nasion, upper incisor, and lower incisor) were obtained by measuring the distance and direction of the landmarks from the MRP, and the two methods were compared statistically. The direction of deviation and the severity of asymmetry found using each method were also compared. The x-coordinates of the four anatomic landmarks all showed a statistically significant difference between the two methods of establishing the MRP. For the nasion and lower incisor, six patients (25.0%) showed a change in the direction of deviation. The severity of asymmetry also changed in 16 patients (66.7%). The results of this study suggest that the locations of midfacial landmarks change significantly according to the method used to establish the MRP.

  10. Glucose phosphorylation is required for Mycobacterium tuberculosis persistence in mice.

    Directory of Open Access Journals (Sweden)

    Joeli Marrero

    2013-01-01

    Full Text Available Mycobacterium tuberculosis (Mtb is thought to preferentially rely on fatty acid metabolism to both establish and maintain chronic infections. Its metabolic network, however, allows efficient co-catabolism of multiple carbon substrates. To gain insight into the importance of carbohydrate substrates for Mtb pathogenesis we evaluated the role of glucose phosphorylation, the first reaction in glycolysis. We discovered that Mtb expresses two functional glucokinases. Mtb required the polyphosphate glucokinase PPGK for normal growth on glucose, while its second glucokinase GLKA was dispensable. (13C-based metabolomic profiling revealed that both enzymes are capable of incorporating glucose into Mtb's central carbon metabolism, with PPGK serving as dominant glucokinase in wild type (wt Mtb. When both glucokinase genes, ppgK and glkA, were deleted from its genome, Mtb was unable to use external glucose as substrate for growth or metabolism. Characterization of the glucokinase mutants in mouse infections demonstrated that glucose phosphorylation is dispensable for establishing infection in mice. Surprisingly, however, the glucokinase double mutant failed to persist normally in lungs, which suggests that Mtb has access to glucose in vivo and relies on glucose phosphorylation to survive during chronic mouse infections.

  11. Genomic homogeneity between Mycobacterium avium subsp. avium and Mycobacterium avium subsp. paratuberculosis belies their divergent growth rates

    Directory of Open Access Journals (Sweden)

    Li Ling-Ling

    2003-05-01

    Full Text Available Abstract Background Mycobacterium avium subspecies avium (M. avium is frequently encountered in the environment, but also causes infections in animals and immunocompromised patients. In contrast, Mycobacterium avium subspecies paratuberculosis (M. paratuberculosis is a slow-growing organism that is the causative agent of Johne's disease in cattle and chronic granulomatous infections in a variety of other ruminant hosts. Yet we show that despite their divergent phenotypes and the diseases they present, the genomes of M. avium and M. paratuberculosis share greater than 97% nucleotide identity over large (25 kb genomic regions analyzed in this study. Results To characterize genome similarity between these two subspecies as well as attempt to understand their different growth rates, we designed oligonucleotide primers from M. avium sequence to amplify 15 minimally overlapping fragments of M. paratuberculosis genomic DNA encompassing the chromosomal origin of replication. These strategies resulted in the successful amplification and sequencing of a contiguous 11-kb fragment containing the putative Mycobacterium paratuberculosis origin of replication (oriC. This fragment contained 11 predicted open reading frames that showed a conserved gene order in the oriC locus when compared with several other Gram-positive bacteria. In addition, a GC skew analysis identified the origin of chromosomal replication which lies between the genes dnaA and dnaN. The presence of multiple DnaA boxes and the ATP-binding site in dnaA were also found in M. paratuberculosis. The strong nucleotide identity of M. avium and M. paratuberculosis in the region surrounding the origin of chromosomal replication led us to compare other areas of these genomes. A DNA homology matrix of 2 million nucleotides from each genome revealed strong synteny with only a few sequences present in one genome but absent in the other. Finally, the 16s rRNA gene from these two subspecies is 100

  12. Establishment of a brain perfusion single photon emission computed tomography database in children by statistical imaging analysis

    International Nuclear Information System (INIS)

    We made a 3-dimensional, 99mTc-ECD brain perfusion single photon emission computed tomography (SPECT) database in children by a statistical analysis. We selected 52 individuals between 1 and 15 years of age whose brain SPECT and brain MRI findings were normal, and divided them into three age groups: 1-5, 6-10 and 11-15 years. By comparing databases obtained for each group, an age-dependent change of regional cerebral blood flow was investigated. The results showed that the relative blood flow increases in the frontal lobe and cerebeller hemisphere and decrease in the occipital lobe with increasing age, findings consistent with those of previous reports. This database enabled us to easily find 3-dimensional brain perfusion abnormality in individual patient by SPECT, and may help elucidate the pathophysiology of many brain disorders. (author)

  13. Establishment of earth tides effect on water level fluctuations in an unconfined hard rock aquifer using spectral analysis

    CERN Document Server

    Maréchal, Jean-Christophe; Ahmed, Shakeel; Lachassagne, Patrick

    2010-01-01

    Short-interval water level measurements using automatic water level recorder in a deep well in an unconfined crystalline rock aquifer at the campus of NGRI, near Hyderabad shows a cyclic fluctuation in the water levels. The observed values clearly show the principal trend due to rainfall recharge. Spectral analysis was carried out to evaluate correlation of the cyclic fluctuation to the synthetic earth tides as well as groundwater withdrawal time series in the surrounding. It was found that these fluctuations have considerably high correlation with earth tides whereas groundwater pumping does not show any significant correlation with water table fluctuations. It is concluded that earth tides cause the fluctuation in the water table. These fluctuations were hitherto unobserved during manual observations made over larger time intervals. It indicates that the unconfined aquifer is characterised by a low porosity.

  14. A Study on establishing Top-Down model - development of Global CGE model for Kyoto mechanism analysis

    Energy Technology Data Exchange (ETDEWEB)

    Cho, Gyeong Lyeong [Korea Energy Economics Institute, Euiwang (Korea)

    1999-07-01

    In Korea, it is true that studies on the Convention on Climate Change is behind compared with the developed countries. As a result, a clear goal on negotiation and a reasonable alternative to persuade other countries were not prepared and a negative and passive attitude of negotiation was showed when entering into negotiations with other countries. However, fortunately, the Convention on Climate Change is not completely reached an agreement. It means that there still remains time to contrive for terms favorable to Korea. To do this, a precise negotiating strategy should be prepared based on a reasonable and systematic study and with diplomatic negotiating power to assist it. Also, it should prepare for a next stage after the negotiation through the analyses of several scenarios. Developing a model for a simulation analysis is absolutely needed to set a more systematic and reasonable negotiation strategy for the Convention on Climate Change. (author). 32 refs., 14 fig., 36 tabs.

  15. Disseminated Mycobacterium gordonae and Mycobacterium mantenii infection with elevated anti-IFN-gamma neutralizing autoantibodies

    NARCIS (Netherlands)

    Hase, I.; Morimoto, K.; Sakagami, T.; Kazumi, Y.; Ishii, Y.; Ingen, J. van

    2015-01-01

    A case of disseminated nontuberculous mycobacteria(l) (NTM) infection in a patient with positive neutralizing anti-interferon-gamma (IFN-gamma) autoantibodies involving bone, bronchus, systemic lymph nodes, and skin is reported. The causative NTMs were two different strains: Mycobacterium gordonae,

  16. Meta analysis on the co-infection between Mycobacterium tuberculosis and HIV/AIDS in China%中国结核分枝杆菌/艾滋病病毒双重感染率分析

    Institute of Scientific and Technical Information of China (English)

    刘姣; 吕冰; 颜艳

    2013-01-01

    Objective To understand and estimate the rate of co-infection between Mycobacterium tuberculosis (M.tuberculosis) and the human immunodeficiency virus (HIV).Methods Based on the related literature regarding Chinese M.tuberculosis and HIV co-infection being published between 2000 and 2012 in PubMed,China National Knowledge Infrastructure (CNKI),Wan Fang Databases and the Chinese Science & Technology Journal Database (VIP),we extracted related information and applied the generic inverse variance model to estimate the following parameters as:co-infection rate,differences on gender,region and subgroups.We also used the STROBE Statement and observational quality evaluation standard to evaluate the quality of literature,and employed sensitivity analyses to evaluate the impact of research quality.Results 39 papers were included in this Meta-analysis,which contained a total sample size of 168 286.The co-infection rate of HIV in TB patients was 0.86% (95%CI:0.80%-0.93%),when using the generic inverse variance model.According to subgroups analyses,at α =0.05 level,men had a higher co-infection rate (1.53%) than women (0.54%) and areas with high infection rate of HIV/AIDS (0.91%) were more commonly seen than low infection rate areas of HIV/AIDS (0.71%) and with significant differences.The co-infection rate of M.tuberculosis in patients infected with HIV was 14.44% (95%CI:13.62%-15.30%).Data from subgroup analyses also showed that at the α=0.05 level,the co-infection rates from hospital samples (28.64%) were higher than those from the population samples' (4.74%).The co-infection rate (15.79%) from the western region was higher than that in the Central and Eastern regions (13.88%and 6.47% respectively),with significant differences.After excluding the papers that only met a few items of the STRODE Statement,results of the Meta-analysis were very close to the original results.Conclusion The co-infection rate of both M.tuberculosis and HIV was

  17. 龟分枝杆菌和脓肿分枝杆菌药物敏感性分析及E试验法应用评价%Drug sensitivity analysis of Mycobacterium chelonae and Mycobacterium abscessus and evaluation of Etest for susceptibility testing

    Institute of Scientific and Technical Information of China (English)

    桂静; 王峰; 洪创跃; 李金莉; 梁静

    2013-01-01

    目的 分析深圳地区龟分枝杆菌和脓肿分枝杆菌耐药谱,评价E试验法检测龟分枝杆菌和脓肿分枝杆菌药物敏感性的临床适用性.方法 用GenoType Mycobacterium CM分枝杆菌菌种鉴定系统筛选16株龟分枝杆菌临床菌株、20株脓肿分枝杆菌临床菌株,分别用微量肉汤稀释法和E试验法检测其对阿米卡星、头孢西丁、环丙沙星、克拉霉素、多西环素、亚胺培南、利奈唑胺、磺胺甲噁唑/甲氧苄啶和妥布霉素9种抗生素的MIC,以微量肉汤稀释法为标准,评价E试验法的MIC值符合率和MIC解释的一致率.两组间率的比较采用x2检验.结果 龟分枝杆菌和脓肿分枝杆菌对阿米卡星和头孢西丁均敏感(36/36),对克拉霉素的耐药率较低(1/36),对环丙沙星、多西环素、亚胺培南和磺胺甲噁唑/甲氧苄啶的耐药率较高(29/36).龟分枝杆菌和脓肿分枝杆菌对利奈唑胺的耐药例数分别为2/16和12/20,对妥布霉素的耐药例数分别为7/16和16/20.E试验法检测9种抗生素的MIC值分布与微量肉汤稀释法MIC值分布的总体符合率为46%(149/324),MIC解释一致率最高的分别为阿米卡星、克拉霉素、多西环素和亚胺培南(35/36),其次为磺胺甲噁唑/甲氧苄啶(34/36),2例(2/36)解释结果有重大误差;一致率较高的为环丙沙星(31/36)和妥布霉素(26/36);一致率最低的为头孢西丁和利奈唑胺(14/36),易导致MIC解释出现假阳性.结论 阿米卡星、头孢西丁和克拉霉素对龟分枝杆菌和脓肿分枝杆菌有良好的体外抗菌作用,利奈唑胺、妥布霉素对部分龟分枝杆菌有良好的体外抗菌作用.除头孢西丁和利奈唑胺外,阿米卡星、克拉霉素、多西环素、环丙沙星和妥布霉素均适用E试验法检测龟分枝杆菌和脓肿分枝杆菌的体外药物敏感性,需慎重采用E试验法检测磺胺甲噁唑/甲氧苄啶的药物敏感性.%Objective To study the drug resistance

  18. Genomics of glycopeptidolipid biosynthesis in Mycobacterium abscessus and M. chelonae

    Directory of Open Access Journals (Sweden)

    Etienne Gilles

    2007-05-01

    Full Text Available Abstract Background The outermost layer of the bacterial surface is of crucial importance because it is in constant interaction with the host. Glycopeptidolipids (GPLs are major surface glycolipids present on various mycobacterial species. In the fast-grower model organism Mycobacterium smegmatis, GPL biosynthesis involves approximately 30 genes all mapping to a single region of 65 kb. Results We have recently sequenced the complete genomes of two fast-growers causing human infections, Mycobacterium abscessus (CIP 104536T and M. chelonae (CIP 104535T. We show here that these two species contain genes corresponding to all those of the M. smegmatis "GPL locus", with extensive conservation of the predicted protein sequences consistent with the production of GPL molecules indistinguishable by biochemical analysis. However, the GPL locus appears to be split into several parts in M. chelonae and M. abscessus. One large cluster (19 genes comprises all genes involved in the synthesis of the tripeptide-aminoalcohol moiety, the glycosylation of the lipopeptide and methylation/acetylation modifications. We provide evidence that a duplicated acetyltransferase (atf1 and atf2 in M. abscessus and M. chelonae has evolved through specialization, being able to transfer one acetyl at once in a sequential manner. There is a second smaller and distant (M. chelonae, 900 kb; M. abscessus, 3 Mb cluster of six genes involved in the synthesis of the fatty acyl moiety and its attachment to the tripeptide-aminoalcohol moiety. The other genes are scattered throughout the genome, including two genes encoding putative regulatory proteins. Conclusion Although these three species produce identical GPL molecules, the organization of GPL genes differ between them, thus constituting species-specific signatures. An hypothesis is that the compact organization of the GPL locus in M. smegmatis represents the ancestral form and that evolution has scattered various pieces throughout the

  19. Establishing mass spectrum of $S=-1$ hyperon resonances via a dynamical coupled-channels analysis of $K^-p$ reactions

    CERN Document Server

    Kamano, Hiroyuki

    2016-01-01

    We report our recent effort for the extraction of resonance parameters (complex pole mass and residues etc.) associated with Lambda* and Sigma* hyperons. This was accomplished via a comprehensive partial-wave analysis of the data for K^- p --> barK N, pi Sigma, pi Lambda, eta Lambda, K Xi reactions from the thresholds up to W=2.1 GeV within a dynamical coupled-channels approach. The results suggest a possible existence of new narrow J^P=3/2^+ \\Lambda resonance with pole mass 1671^{+2}_{-8} -i(5^{+11}_{-2}) MeV, located close to the eta Lambda threshold. This resonance is found to be responsible for reproducing the data for K^-p --> eta Lambda differential cross sections near the threshold, and thus the data seem favor its existence. The extracted poles for J^P=1/2^- Lambda resonances below the barK N threshold, including Lambda(1405), are also presented.

  20. Instrumental neutron activation analysis as an analytical tool supporting the establishment of guidelines and databases for workers' health awareness programmes

    International Nuclear Information System (INIS)

    dangerous diseases that are easily identified. The main problem is that the majority of workers are exposed to low levels of toxic chemicals that can be lethal in the long term, owing to chronic diseases. Most often the onset of the diseases goes unnoticed, and the presence of a lung cancer or heart disease is attributed to non-occupational causes. As a result, these cases of illness do not become part of the compiled data. Besides, there is no specialized and complete literature concerning occupational aetiology, nor is there an evaluation of the onset of disease linked to long term exposure to low levels of toxic agents. With the aim of giving support to the Workers' Health Awareness Programme of the Secretaria Municipal de Saude (Municipal Department of Health) of Belo Horizonte, capital of Minas Gerais state, an assessment was done in galvanizing factories by means of airborne particulate matter collected in air filters and in hair and toenails as biomonitors. This project was approved by the Ethics Committee of the Federal University of Minas Gerais, COEP-UFMG. All research involving human beings has to be submitted to this committee in order to protect the population studied. The k0 instrumental neutron activation analysis (INAA) technique was chosen to be applied for the determination of elements in air filters and in hair and toenail samples, as it can determine several elements in the same sample almost simultaneously, with a low detection limit and without any chemical procedure as is required in the majority of non-nuclear techniques, and also because it requires only a small amount of sample for the analysis. The k0 method was applied to all samples, demonstrating its quality as a versatile technique, and it was confirmed to be one of the most advantageous and suitable nuclear analytical techniques

  1. Validation of a Multiplex Real-Time PCR Assay for Detection of Mycobacterium spp., Mycobacterium tuberculosis Complex, and Mycobacterium avium Complex Directly from Clinical Samples by Use of the BD Max Open System.

    Science.gov (United States)

    Rocchetti, Talita T; Silbert, Suzane; Gostnell, Alicia; Kubasek, Carly; Widen, Raymond

    2016-06-01

    A multiplex real-time PCR was validated on the BD Max open system to detect different Mycobacterium tuberculosis complex, Mycobacterium avium complex, and Mycobacterium spp. directly from clinical samples. The PCR results were compared to those with traditional cultures. The multiplex PCR assay was found to be a specific and sensitive method for the rapid detection of mycobacteria directly from clinical specimens.

  2. Mycobacterium fortuitum abdominal wall abscesses following liposuction

    Directory of Open Access Journals (Sweden)

    Al Soub Hussam

    2008-01-01

    Full Text Available We describe here a case of abdominal abscesses due to Mycobacterium fortuitum following liposuction. The abscesses developed three months after the procedure and diagnosis was delayed for five months. The clues for diagnosis were persistent pus discharge in spite of broad spectrum antibiotics and failure to grow any organisms on routine culture. This condition has been rarely reported; however, the increasing number of liposuction procedures done and awareness among physicians will probably result in the identification of more cases. Combination antibiotic therapy with surgical drainage in more extensive diseases is essential for cure.

  3. Dormancy models for Mycobacterium tuberculosis: A minireview.

    Science.gov (United States)

    Alnimr, Amani M

    2015-01-01

    Dormancy models for Mycobacterium tuberculosis play important roles in understanding various aspects of tuberculosis pathogenesis and in the testing of novel therapeutic regimens. By simulating the latent tuberculosis infection, in which the bacteria exist in a non-replicative state, the models demonstrate reduced susceptibility to antimycobacterial agents. This minireview outlines the models available for simulating latent tuberculosis both in vitro and in several animal species. Additionally, this minireview discusses the advantages and disadvantages of these models for investigating the bacterial subpopulations and susceptibilities to sterilization by various antituberculosis drugs. PMID:26413043

  4. Dormancy models for Mycobacterium tuberculosis: A minireview

    Directory of Open Access Journals (Sweden)

    Amani M. Alnimr

    2015-09-01

    Full Text Available Dormancy models for Mycobacterium tuberculosis play important roles in understanding various aspects of tuberculosis pathogenesis and in the testing of novel therapeutic regimens. By simulating the latent tuberculosis infection, in which the bacteria exist in a non-replicative state, the models demonstrate reduced susceptibility to antimycobacterial agents. This minireview outlines the models available for simulating latent tuberculosis both in vitro and in several animal species. Additionally, this minireview discusses the advantages and disadvantages of these models for investigating the bacterial subpopulations and susceptibilities to sterilization by various antituberculosis drugs.

  5. The Evolving Mcart Multimodal Imaging Core: Establishing a Protocol for Computed Tomography and Echocardiography in the Rhesus Macaque to Perform Longitudinal Analysis of Radiation-Induced Organ Injury.

    Science.gov (United States)

    de Faria, Eduardo B; Barrow, Kory R; Ruehle, Bradley T; Parker, Jordan T; Swartz, Elisa; Taylor-Howell, Cheryl; Kieta, Kaitlyn M; Lees, Cynthia J; Sleeper, Meg M; Dobbin, Travis; Baron, Adam D; Mohindra, Pranshu; MacVittie, Thomas J

    2015-11-01

    Computed Tomography (CT) and Echocardiography (EC) are two imaging modalities that produce critical longitudinal data that can be analyzed for radiation-induced organ-specific injury to the lung and heart. The Medical Countermeasures Against Radiological Threats (MCART) consortium has a well established animal model research platform that includes nonhuman primate (NHP) models of the acute radiation syndrome and the delayed effects of acute radiation exposure. These models call for a definition of the latency, incidence, severity, duration, and resolution of different organ-specific radiation-induced subsyndromes. The pulmonary subsyndromes and cardiac effects are a pair of interdependent syndromes impacted by exposure to potentially lethal doses of radiation. Establishing a connection between these will reveal important information about their interaction and progression of injury and recovery. Herein, the authors demonstrate the use of CT and EC data in the rhesus macaque models to define delayed organ injury, thereby establishing: a) consistent and reliable methodology to assess radiation-induced damage to the lung and heart; b) an extensive database in normal age-matched NHP for key primary and secondary endpoints; c) identified problematic variables in imaging techniques and proposed solutions to maintain data integrity; and d) initiated longitudinal analysis of potentially lethal radiation-induced damage to the lung and heart.

  6. Direct identification and discernment of Mycobacterium avium and Mycobacterium intracellulare using a real-time RNA isothermal amplification and detection method.

    Science.gov (United States)

    Cui, Zhenling; Li, Yuanyuan; Cheng, Song; Yang, Hua; Lu, Junmei; Zhu, Honglei; Hu, Zhongyi

    2015-12-01

    The purpose of this work was to establish a real-time simultaneous amplification and testing method for identification and discernment of Mycobacterium avium and Mycobacterium intracellulare (SAT-MAC assay) and to evaluate the efficiency with which this method can detect isolated strains and clinical sputum specimens. The specific 16S rRNA sequences of M. avium and M. intracellulare were used as targets to design RNA probes and a reverse transcription primer containing T7 promoter. RNA isothermal amplification and real-time fluorescence detection were performed at 42 °C. SAT-MAC assay, culture tests on Lowenstein-Jensen (L-J) culture medium and PCR-sequencing were used to test the clinical isolated strains and sputum specimens. The limit of detection (LOD) of M. avium and M. intracellulare by SAT-MAC was found to be 30 CFU/mL and 20 CFU/mL. SAT-MAC showed high specificity in 21 species of mycobacteria standard strains and 5 species of non-mycobacteria bacteria. Using PCR-sequencing as the reference method, both rates of SAT-MAC assay for identifying M. avium and M. intracellulare from clinical isolates were 100% (259/259). Consistent with the results of L-J culture combined PCR-sequencing, the coincidence rate of SAT-MAC assay in clinical sputum specimens was 100% (369/369) for M. avium and 99.19% (366/369) for Mycobacterium intracellular. The SAT-MAC assay can identify and distinguish M. avium and M. intracellulare rapidly and accurately. It may be suitable for use in clinical microbiology laboratories.

  7. 荧光定量PCR技术检测结核杆菌临床应用分析%Analysis on the Clinical Application of Fluorescence Quantitative PCR Assay in Detection of Mycobacterium Tuberculosis

    Institute of Scientific and Technical Information of China (English)

    王雅宁

    2012-01-01

    Objective To investigate the clinical application value of fluorescence quantitative PCR (FQ — PCR) technigue in the detection of mycobacterium tuberculosis. Methods Sixty — eight patients clinically diagnosed as pulmonary tuberculosis in the First Hospital of Baoding, Hebei Province, were chosen as the research objects. Mycobacterium tuberculosis in peripheral blood and one portion of sputum was determined by FQ- PCR. Another portion of sputum was subject to sputum smear. Clinical diagnosis was used as the gold standard. The results were analyzed by χ2 test. Results The positive rate of mycobacterium tuberculosis detected by sputum smear, sputum FQ- PCR and peripheral blood FQ- PCR in 68 patients with pulmonary tuberculosis was 14.7%, 41.2% and 44.1%, respectively. The positive rates by FQ- PCR were significantly higher than that by sputum smear (P < 0.05). The positive rates by sputum FQ - PCR and peripheral blood FQ - PCR in 58 cases with negative detection results by sputum smear was 31.0% and 41.3% . The coincidence rate by FQ - PCR in sputum and in paired peripheral blood was 44. 1 % . Conclusions Fluorescence quantitative PCR assay is superior to conventional sputum smear in mycobacte rium tuber culosis detection. It can confirm and identify the positive results of sputum smear. To simultaneously conduct FQ-PCR assay in sputum and peripheral blood can have complementary effect and improve the detection accuracy.%目的 探讨荧光定量PCR技术在检测结核杆菌的临床应用价值. 方法 将本院临床诊断确诊为肺结核的68例患者为研究对象,采用荧光PCR技术测定两组痰液及外周血中的结核杆菌,以临床诊断结果为金标准,采用x2检验对结果进行分析比较. 结果 68例结核病患者中痰涂片、痰定量PCR、外周血定量PCR检测的阳性率为14.7%、41.2%、44.1%,定量PCR检测的阳性率显著高于痰涂片(P<0.05).痰涂片阴性的58例患者中痰及外周

  8. 实时荧光定量 PCR法分析结核分枝杆菌对异烟肼耐药的分析%Analysis of Mycobacterium tuberculosis resistant to isoniazid by real-time quantitative PCR

    Institute of Scientific and Technical Information of China (English)

    孙京涛; 孙丽娜

    2016-01-01

    目的:利用实时荧光定量PCR技术快速检测异烟肼耐药结核分枝杆菌。方法收集到医院就诊的结核病疑似患者痰液样本,提取痰液样本的总DNA,利用实时荧光定量PCR( Real-time PCR)技术对结核分枝杆菌感染进行快速筛查,并与传统药敏试验进行比较,对两者的灵敏度、特异性、一致性进行比较分析。结果检测346例结核病人临床分离培养样本,药敏试验检出257例异烟肼敏感标本,101例异烟肼耐药标本;实时荧光定量PCR 法共检测出异烟肼敏感和耐药标本225例98例,灵敏度为86.64%,特异性为93.92%,一致率为93.12%。结论跟传统药物敏感性实验相比,实时荧光定量PCR法检测速度快速、特异性强、灵敏度较高,可用于结核分枝杆菌耐异烟肼突变的快速检测,适于耐多药结核病的快速筛查。%Objective To rapid detect isoniazid resistant Mycobacterium tuberculosis by using real -time flu-orescence quantitative PCR technology .Methods The sputum samples of suspected TB patients to extract total DNA by real-time fluorescence quantitative PCR technique for rapid screening of Mycobacterium tuberculosis infection , and then it was compared with conventional drug susceptibility test in sensitivity , specificity , and consistency .Results There were 257 cases of isoniazid sensitive samples and 101 cases of isoniazid resistant samples by drug sensitivity test, and 225 cases of isoniazid sensitive samples and 98 cases of isoniazid resistant samples by real-time fluorescence quantitative PCR.The sensitivity was 86.64%, the specificity was 93.92%, and the concordance was 93.12%. Conclusion Compared with the traditional drug sensitivity test , real-time fluorescence quantitative PCR method has the advantages of quick detection and high specificity and sensitivity for Mycobacterium tuberculosis isoniazid resistant mutation detection , and it is also suitable for rapid screening

  9. Association between Mycobacterium avium subsp. paratuberculosis infection and culling in dairy cattle herds

    Directory of Open Access Journals (Sweden)

    R Arrazuría

    2014-01-01

    Full Text Available The present study was designed to analyse the causes for culling in dairy herds with different Mycobacterium avium subsp. paratuberculosis infection status and to compare these causes with those observed over the general dairy cattle population. During 2009, causes for culling were registered in two different groups of farms: (1 farms with seropositive cows for three consecutive years (2007-2009 but where Mycobacterium avium subsp. paratuberculosis has not been isolated from any of the fecal samples collected and (2 farms with Mycobacterium avium subsp. paratuberculosis seropositive cows for three consecutive years (2007-2009 and where the bacteria has been isolated from at least one fecal sample. Causes for animal loss were compared between both groups and between them and the general dairy cattle population by means of regression analysis. The distribution of culling reasons was different between infected herds (both bacteriologically positive and negative and the general population. The percentage of losses seemed to be higher in infected herds from the first parity on. The most remarkable difference among groups was observed in losses due to "death/urgent slaughter".

  10. Re-establishing dignity

    DEFF Research Database (Denmark)

    Høy, Bente; Hall, E.O.C

    2012-01-01

    nurses’ experiences of caring for older hospital patients. The study was based on Benner and Wrubel’s phenomenological notion that caring as a special kind of involvement, a grasp of a situation in terms of its meaning, is primary in nursing. A qualitative secondary analysis of data from an interview...... study with 29 nurses and nurse assistants was performed following Van Manen’s hermeneutic phenomenology. Findings show that caring mainly concerns ‘re-establishing dignity’ especially through ‘seeing the patient as a unique person’, ‘assisting in getting rid of the bed’ and ‘supporting patient...

  11. Establishment of a System for Monitoring and Control of Cross Contamination in the Laboratory of Microbiological Analysis of Food during 2009

    Directory of Open Access Journals (Sweden)

    Corpa- Iguarán Eduardo Javid

    2012-03-01

    Full Text Available Within the activities for quality control in the laboratory, the final results of a particular analyteare now recognized as intermediates, due to the relevance given to quality assurance, as the ultimategoal of programs for quality management. This concept makes it necessary to establishmentof integral tools, to detect events such as cross-contamination, and measures to avoid affectingthe analysis method. Objective: to established a system for monitoring and control of crosscontaminationin the laboratory of microbiological analysis of food. Materials and methods:Flowcharts were developed to control the proceedings on the populations of aerobe mesophilicsand molds originated from pollution in the environment, surfaces, sterile material and culturemedia, which included a decision tree designed to perform control actions based on tolerance intervals,which were established as objective tool to decision-making leading to normalize countsof microbial populations in question. Results: Warning limits stricter were obtained for aerobicmesophilic and mold populations in the different controls, except for environment of media preparationand the corresponding for sterile material. Conclusion: The process developed allowedto complement the system of internal quality control in the laboratory, to provide of an objectivetool for closing nonconformities because of cross-contamination.

  12. Establishment of water source discrimination model in coal mine by using hydrogeochemistry and statistical analysis: a case study from Renlou Coal Mine in northern Anhui Province, China

    Institute of Scientific and Technical Information of China (English)

    SUN Lin-hua; GUI He-rong

    2012-01-01

    The demand for energy consumption promotes to find more coal in deep underground up to 1000 m and brings more serious situation of water disaster.As one of the major methods for water disaster control,hydrogeochemistry attracts a series of studies related to water source discrimination.In this paper,a simple method for constructing the water source discrimination model based on major ions and multivariate statistical analysis was reported using the following procedures:① collection of data and interpretation,② analysis of controlling factors based on the chemical composition of groundwater,③ "pure" sample chosen,and ④ discrimination model establishment.After the processes,two functions and a diagram were established for three aquifers (the Quaternary,Coal bearing,and Taiyuan Fm.) from the Renlou Coal Mine in northern Anhui Province,China.The method can be applied in almost all coal mines and can be used for evaluating the contribution ratios if the water is collected from a mixing source.

  13. Volatile emissions from Mycobacterium avium subsp. paratuberculosis mirror bacterial growth and enable distinction of different strains.

    Directory of Open Access Journals (Sweden)

    Phillip Trefz

    Full Text Available Control of paratuberculosis in livestock is hampered by the low sensitivity of established direct and indirect diagnostic methods. Like other bacteria, Mycobacterium avium subsp. paratuberculosis (MAP emits volatile organic compounds (VOCs. Differences of VOC patterns in breath and feces of infected and not infected animals were described in first pilot experiments but detailed information on potential marker substances is missing. This study was intended to look for characteristic volatile substances in the headspace of cultures of different MAP strains and to find out how the emission of VOCs was affected by density of bacterial growth. One laboratory adapted and four field strains, three of MAP C-type and one MAP S-type were cultivated on Herrold's egg yolk medium in dilutions of 10(-0, 10(-2, 10(-4 and 10(-6. Volatile substances were pre-concentrated from the headspace over the MAP cultures by means of Solid Phase Micro Extraction (SPME, thermally desorbed from the SPME fibers and separated and identified by means of GC-MS. Out of the large number of compounds found in the headspace over MAP cultures, 34 volatile marker substances could be identified as potential biomarkers for growth and metabolic activity. All five MAP strains could clearly be distinguished from blank culture media by means of emission patterns based on these 34 substances. In addition, patterns of volatiles emitted by the reference strain were significantly different from the field strains. Headspace concentrations of 2-ethylfuran, 2-methylfuran, 3-methylfuran, 2-pentylfuran, ethyl acetate, 1-methyl-1-H-pyrrole and dimethyldisulfide varied with density of bacterial growth. Analysis of VOCs emitted from mycobacterial cultures can be used to identify bacterial growth and, in addition, to differentiate between different bacterial strains. VOC emission patterns may be used to approximate bacterial growth density. In a perspective volatile marker substances could be used to

  14. Mycobacterium tuberculosis is resistant to streptolydigin.

    Science.gov (United States)

    Speer, Alexander; Rowland, Jennifer L; Niederweis, Michael

    2013-07-01

    Drug resistant strains of Mycobacterium tuberculosis (Mtb) undermine tuberculosis (TB) control. Streptolydigin is a broadly effective antibiotic which inhibits RNA polymerase, similarly to rifampicin, a key drug in current TB chemotherapeutic regimens. Due to a vastly improved chemical synthesis streptolydigin and derivatives are being promoted as putative TB drugs. The microplate Alamar Blue assay revealed that Streptococcus salivarius and Mycobacterium smegmatis were susceptible to streptolydigin with minimum inhibitory concentrations (MICs) of 1.6 mg/L and 6.25 mg/L, respectively. By contrast, the MICs of streptolydigin and two derivatives, streptolydiginone and dihydrostreptolydigin, against Mtb were ≥ 100 mg/L demonstrating that Mtb is resistant to streptolydigin in contrast to previous reports. Further, a porin mutant of M. smegmatis is resistant to streptolydigin indicating that porins mediate uptake of streptolydigin across the outer membrane. Since the RNA polymerase is a validated drug target in Mtb and porins are required for susceptibility of M. smegmatis, the absence of MspA-like porins probably contributes to the resistance of Mtb to streptolydigin. This study shows that streptolydigin is not a suitable drug in TB treatment regimens. PMID:23591156

  15. Porins increase copper susceptibility of Mycobacterium tuberculosis.

    Science.gov (United States)

    Speer, Alexander; Rowland, Jennifer L; Haeili, Mehri; Niederweis, Michael; Wolschendorf, Frank

    2013-11-01

    Copper resistance mechanisms are crucial for many pathogenic bacteria, including Mycobacterium tuberculosis, during infection because the innate immune system utilizes copper ions to kill bacterial intruders. Despite several studies detailing responses of mycobacteria to copper, the pathways by which copper ions cross the mycobacterial cell envelope are unknown. Deletion of porin genes in Mycobacterium smegmatis leads to a severe growth defect on trace copper medium but simultaneously increases tolerance for copper at elevated concentrations, indicating that porins mediate copper uptake across the outer membrane. Heterologous expression of the mycobacterial porin gene mspA reduced growth of M. tuberculosis in the presence of 2.5 μM copper by 40% and completely suppressed growth at 15 μM copper, while wild-type M. tuberculosis reached its normal cell density at that copper concentration. Moreover, the polyamine spermine, a known inhibitor of porin activity in Gram-negative bacteria, enhanced tolerance of M. tuberculosis for copper, suggesting that copper ions utilize endogenous outer membrane channel proteins of M. tuberculosis to gain access to interior cellular compartments. In summary, these findings highlight the outer membrane as the first barrier against copper ions and the role of porins in mediating copper uptake in M. smegmatis and M. tuberculosis. PMID:24013632

  16. Images of mycobacterium for nuclear reactions

    International Nuclear Information System (INIS)

    According to the World Health Organization (WHO) tuberculosis is responsible for 2.9 million deaths annually worldwide. The necessity for optimizing time to detect the tuberculosis bacillus (mycobacterium tuberculosis) in the sputum samples of affected individuals (TB patients) led to the development of a methodology based on the doping with boron of the bacillus, submission of the samples to thermal neutron beam and ionizing particles, generating nuclear reactions of the types: 10B (n,α)7Li and 10 B(α, p)13C. Images of these bacilli are obtained by means of the nuclear tracks produced in the CR-39 detector for particles products of these nuclear reactions, α and p. When the CR-39 is submitted to a chemical attack the traces are developed and the images of the microorganisms registered in the detector can be observed with a conventional light microscope, characterizing them by morphology. The use of this methodology results in images of the mycobacterium tuberculosis becoming more defined and enlarged than those obtained by bacilloscopy, in which the sample is submitted to the method of coloration of Ziehl-Neelsen (ZN) and observed in light microscopy. (author)

  17. Mycobacterium abscessus phospholipase C expression is induced during coculture within amoebae and enhances M. abscessus virulence in mice.

    Science.gov (United States)

    Bakala N'Goma, Jean Claude; Le Moigne, Vincent; Soismier, Nathalie; Laencina, Laura; Le Chevalier, Fabien; Roux, Anne-Laure; Poncin, Isabelle; Serveau-Avesque, Carole; Rottman, Martin; Gaillard, Jean-Louis; Etienne, Gilles; Brosch, Roland; Herrmann, Jean-Louis; Canaan, Stéphane; Girard-Misguich, Fabienne

    2015-02-01

    Mycobacterium abscessus is a pathogenic, rapidly growing mycobacterium involved in pulmonary and cutaneo-mucous infections worldwide, to which cystic fibrosis patients are exquisitely susceptible. The analysis of the genome sequence of M. abscessus showed that this bacterium is endowed with the metabolic pathways typically found in environmental microorganisms that come into contact with soil, plants, and aquatic environments, where free-living amoebae are frequently present. M. abscessus also contains several genes that are characteristically found only in pathogenic bacteria. One of them is MAB_0555, encoding a putative phospholipase C (PLC) that is absent from most other rapidly growing mycobacteria, including Mycobacterium chelonae and Mycobacterium smegmatis. Here, we report that purified recombinant M. abscessus PLC is highly cytotoxic to mouse macrophages, presumably due to hydrolysis of membrane phospholipids. We further showed by constructing and using an M. abscessus PLC knockout mutant that loss of PLC activity is deleterious to M. abscessus intracellular survival in amoebae. The importance of PLC is further supported by the fact that M. abscessus PLC was found to be expressed only in amoebae. Aerosol challenge of mice with M. abscessus strains that were precultured in amoebae enhanced M. abscessus lung infectivity relative to M. abscessus grown in broth culture. Our study underlines the importance of PLC for the virulence of M. abscessus. Despite the difficulties of isolating M. abscessus from environmental sources, our findings suggest that M. abscessus has evolved in close contact with environmental protozoa, which supports the argument that amoebae may contribute to the virulence of opportunistic mycobacteria.

  18. Lymphatic endothelial cells are a replicative niche for Mycobacterium tuberculosis

    Science.gov (United States)

    Lerner, Thomas R.; de Souza Carvalho-Wodarz, Cristiane; Repnik, Urska; Russell, Matthew R.G.; Borel, Sophie; Diedrich, Collin R.; Rohde, Manfred; Wainwright, Helen; Collinson, Lucy M.; Wilkinson, Robert J.; Griffiths, Gareth; Gutierrez, Maximiliano G.

    2016-01-01

    In extrapulmonary tuberculosis, the most common site of infection is within the lymphatic system, and there is growing recognition that lymphatic endothelial cells (LECs) are involved in immune function. Here, we identified LECs, which line the lymphatic vessels, as a niche for Mycobacterium tuberculosis in the lymph nodes of patients with tuberculosis. In cultured primary human LECs (hLECs), we determined that M. tuberculosis replicates both in the cytosol and within autophagosomes, but the bacteria failed to replicate when the virulence locus RD1 was deleted. Activation by IFN-γ induced a cell-autonomous response in hLECs via autophagy and NO production that restricted M. tuberculosis growth. Thus, depending on the activation status of LECs, autophagy can both promote and restrict replication. Together, these findings reveal a previously unrecognized role for hLECs and autophagy in tuberculosis pathogenesis and suggest that hLECs are a potential niche for M. tuberculosis that allows establishment of persistent infection in lymph nodes. PMID:26901813

  19. Bioluminescence for assessing drug potency against nonreplicating Mycobacterium tuberculosis.

    Science.gov (United States)

    Vocat, Anthony; Hartkoorn, Ruben C; Lechartier, Benoit; Zhang, Ming; Dhar, Neeraj; Cole, Stewart T; Sala, Claudia

    2015-07-01

    Targeting dormant Mycobacterium tuberculosis represents a challenge to antituberculosis drug discovery programs. We previously reported and validated the use of the streptomycin (STR)-dependent M. tuberculosis 18b strain as a tool for assessing drug potency against nonreplicating bacteria both in vitro and in vivo. In this study, we generated a luminescent 18b strain, named 18b-Lux, by transforming the bacteria with a vector expressing the luxCDABE operon from Photorhabdus luminescens. Luciferase expression was demonstrated under replicating conditions, and, more importantly, luminescence levels significantly above background were detected following STR removal. The sensitivity of STR-starved 18b-Lux to approved and candidate antituberculosis therapeutic agents was evaluated by means of a luciferase assay in a 96-well format. Results mirrored the data obtained with the standard resazurin reduction microplate assay, and the luminescence readout allowed time course assessments of drug efficacy in vitro. Specifically, we proved that bedaquiline, the rifamycins, and sutezolid displayed time-dependent activity against dormant bacteria, while pyrazinamide and SQ109 showed bactericidal effects at the highest concentrations tested. Overall, we established the optimal conditions for an inexpensive, simple, and very sensitive assay with great potential for future applications. PMID:25896710

  20. An acidic sphingomyelinase Type C activity from Mycobacterium tuberculosis.

    Science.gov (United States)

    Castro-Garza, Jorge; González-Salazar, Francisco; Quinn, Frederick D; Karls, Russell K; De La Garza-Salinas, Laura Hermila; Guzmán-de la Garza, Francisco J; Vargas-Villarreal, Javier

    2016-01-01

    Sphingomyelinases (SMases) catalyze the hydrolysis of sphingomyelin to ceramide and phosphorylcholine. Sphingolipids are recognized as diverse and dynamic regulators of a multitude of cellular processes mediating cell cycle control, differentiation, stress response, cell migration, adhesion, and apoptosis. Bacterial SMases are virulence factors for several species of pathogens. Whole cell extracts of Mycobacterium tuberculosis strains H37Rv and CDC1551 were assayed using [N-methyl-(14)C]-sphingomyelin as substrate. Acidic Zn(2+)-dependent SMase activity was identified in both strains. Peak SMase activity was observed at pH 5.5. Interestingly, overall SMase activity levels from CDC1551 extracts are approximately 1/3 of those of H37Rv. The presence of exogenous SMase produced by M. tuberculosis during infection may interfere with the normal host inflammatory response thus allowing the establishment of infection and disease development. This Type C activity is different from previously identified M. tuberculosis SMases. Defining the biochemical characteristics of M. tuberculosis SMases helps to elucidate the roles that these enzymes play during infection and disease. PMID:26948102

  1. ESX-5-deficient Mycobacterium marinum is hypervirulent in adult zebrafish

    KAUST Repository

    Weerdenburg, Eveline M.

    2012-02-15

    ESX-5 is a mycobacterial type VII protein secretion system responsible for transport of numerous PE and PPE proteins. It is involved in the induction of host cell death and modulation of the cytokine response in vitro. In this work, we studied the effects of ESX-5 in embryonic and adult zebrafish using Mycobacterium marinum. We found that ESX-5-deficient M.marinum was slightly attenuated in zebrafish embryos. Surprisingly, the same mutant showed highly increased virulence in adult zebrafish, characterized by increased bacterial loads and early onset of granuloma formation with rapid development of necrotic centres. This early onset of granuloma formation was accompanied by an increased expression of pro-inflammatory cytokines and tissue remodelling genes in zebrafish infected with the ESX-5 mutant. Experiments using RAG-1-deficient zebrafish showed that the increased virulence of the ESX-5 mutant was not dependent on the adaptive immune system. Mixed infection experiments with wild-type and ESX-5 mutant bacteria showed that the latter had a specific advantage in adult zebrafish and outcompeted wild-type bacteria. Together our experiments indicate that ESX-5-mediated protein secretion is used by M.marinum to establish a moderate and persistent infection. © 2012 Blackwell Publishing Ltd.

  2. Dominant modern sublineages and a new modern sublineage of Mycobacterium tuberculosis Beijing family clinical isolates in Heilongjiang Province, China.

    Science.gov (United States)

    Li, Di; Dong, Cai-Bo; Cui, Jia-Yi; Nakajima, Chie; Zhang, Chun-Lei; Pan, Xin-Ling; Sun, Gao-Xiang; Dai, En-Yu; Suzuki, Yasuhiko; Zhuang, Min; Ling, Hong

    2014-10-01

    Mycobacterium tuberculosis Beijing family includes a variety of sublineages. Knowledge of the distribution of a certain sublineage of the Beijing family may help to understand the mechanisms of its rapid spread and to establish an association between a certain genotype and the disease outcome. We have previously found that M. tuberculosis Beijing family clinical isolates represent approximately 90% of the clinical isolates from Heilongjiang Province, China. To clarify the distribution of M. tuberculosis Beijing family sublineages in Heilongjiang Province, China and to investigate the regularity rule for their evolution, we examined single nucleotide polymorphisms (SNPs) of 250 M. tuberculosis Beijing family clinical isolates using 10 SNP loci that have been identified as appropriate for defining Beijing sublineages. After determining the sequence type (ST) of each isolate, the sublineages of all M. tuberculosis Beijing family isolates were determined, and phylogenetic analysis was performed. We found that 9 out of the 10 SNP loci displayed polymorphisms, but locus 1548149 did not. In total, 92.8% of the isolates in Heilongjiang Province are modern sublineages. ST10 is the most prevalent sublineage (ST10 and ST22 accounted for 63.2% and 23.6% of all the Beijing family isolates, respectively). A new ST, accounting for 4% of the Beijing family isolates in this area, was found for the first time. Each new ST isolate showed a unique VNTR pattern, and none were clustered. The present findings suggest that controlling the spread of these modern sublineages is important in Heilongjiang Province and in China.

  3. Sensitivity and specificity of Cobas TaqMan MTB real-time polymerase chain reaction for culture-proven Mycobacterium tuberculosis: meta-analysis of 26999 specimens from 17 Studies.

    Science.gov (United States)

    Horita, Nobuyuki; Yamamoto, Masaki; Sato, Takashi; Tsukahara, Toshinori; Nagakura, Hideyuki; Tashiro, Ken; Shibata, Yuji; Watanabe, Hiroki; Nagai, Kenjiro; Nakashima, Kentaro; Ushio, Ryota; Ikeda, Misako; Sakamaki, Kentaro; Yoshiyama, Takashi; Kaneko, Takeshi

    2015-12-09

    Since 2010, studies on the diagnostic accuracy of COBAS TaqMan MTB (CTM) have been frequently reported with an unignorable discrepancy. The key inclusion criterion for this systematic review was original studies that could provide sufficient data for calculating the sensitivity and the specificity of CTM for M tuberculosis (TB) or M tuberculosis complex. The reference test was Mycobacterium culture. We used bivariate model for meta-analyses. Of the 201 candidate articles, we finally identified 17 eligible articles.Concerning the respiratory specimens, 1900 culture positive specimens and 20983 culture negative specimens from 15 studies were assessed. This provided the summary estimate sensitivity of 0.808 (95% CI 0.758-0.850) and the summary estimate specificity of 0.990 (95% CI 0.981-0.994). The area under curve was 0.956. The diagnostic odds ratio was 459 (95% CI 261-805, I(2) 26%). For the smear positive respiratory specimens, the sensitivity was 0.952 (95% CI 0.926-0.969) and the specificity was 0.916 (95% CI 0.797-0.968). For the smear negative respiratory specimens, the sensitivity and the specificity were 0.600 (95% CI 0.459-0.726) and 0.989 (95% CI 0.981-0.993), respectively. The diagnostic accuracy was poorer for the non-respiratory specimens, than for the respiratory specimens, but was acceptable. We believe that the information obtained from this study will aid physicians' decision making.

  4. Novel multiplex real-time PCR diagnostic assay for identification and differentiation of Mycobacterium tuberculosis, Mycobacterium canettii, and Mycobacterium tuberculosis complex strains.

    Science.gov (United States)

    Reddington, Kate; O'Grady, Justin; Dorai-Raj, Siobhan; Maher, Majella; van Soolingen, Dick; Barry, Thomas

    2011-02-01

    Tuberculosis (TB) in humans is caused by members of the Mycobacterium tuberculosis complex (MTC). Rapid detection of the MTC is necessary for the timely initiation of antibiotic treatment, while differentiation between members of the complex may be important to guide the appropriate antibiotic treatment and provide epidemiological information. In this study, a multiplex real-time PCR diagnostics assay using novel molecular targets was designed to identify the MTC while simultaneously differentiating between M. tuberculosis and M. canettii. The lepA gene was targeted for the detection of members of the MTC, the wbbl1 gene was used for the differentiation of M. tuberculosis and M. canettii from the remainder of the complex, and a unique region of the M. canettii genome, a possible novel region of difference (RD), was targeted for the specific identification of M. canettii. The multiplex real-time PCR assay was tested using 125 bacterial strains (64 MTC isolates, 44 nontuberculosis mycobacteria [NTM], and 17 other bacteria). The assay was determined to be 100% specific for the mycobacteria tested. Limits of detection of 2.2, 2.17, and 0.73 cell equivalents were determined for M. tuberculosis/M. canettii, the MTC, and M. canettii, respectively, using probit regression analysis. Further validation of this diagnostics assay, using clinical samples, should demonstrate its potential for the rapid, accurate, and sensitive diagnosis of TB caused by M. tuberculosis, M. canettii, and the other members of the MTC.

  5. Genome-wide discovery of small RNAs in Mycobacterium tuberculosis.

    Directory of Open Access Journals (Sweden)

    Paolo Miotto

    Full Text Available Only few small RNAs (sRNAs have been characterized in Mycobacterium tuberculosis and their role in regulatory networks is still poorly understood. Here we report a genome-wide characterization of sRNAs in M. tuberculosis integrating experimental and computational analyses. Global RNA-seq analysis of exponentially growing cultures of M. tuberculosis H37Rv had previously identified 1373 sRNA species. In the present report we show that 258 (19% of these were also identified by microarray expression. This set included 22 intergenic sRNAs, 84 sRNAs mapping within 5'/3' UTRs, and 152 antisense sRNAs. Analysis of promoter and terminator consensus sequences identified sigma A promoter consensus sequences for 121 sRNAs (47%, terminator consensus motifs for 22 sRNAs (8.5%, and both motifs for 35 sRNAs (14%. Additionally, 20/23 candidates were visualized by Northern blot analysis and 5' end mapping by primer extension confirmed the RNA-seq data. We also used a computational approach utilizing functional enrichment to identify the pathways targeted by sRNA regulation. We found that antisense sRNAs preferentially regulated transcription of membrane-bound proteins. Genes putatively regulated by novel cis-encoded sRNAs were enriched for two-component systems and for functional pathways involved in hydrogen transport on the membrane.

  6. Multiplex-PCR for differentiation of Mycobacterium bovis from Mycobacterium tuberculosis complex.

    Science.gov (United States)

    Spositto, F L E; Campanerut, P A Z; Ghiraldi, L D; Leite, C Q F; Hirata, M H; Hirata, R D C; Siqueira, V L D; Cardoso, R Fressatti

    2014-01-01

    We evaluated a multiplex-PCR to differentiate Mycobacterium bovis from M. tuberculosis Complex (MTC) by one step amplification based on simultaneous detection of pncA 169 C > G change in M. bovis and the IS6110 present in MTC species. Our findings showed the proposed multiplex-PCR is a very useful tool for complementation in differentiating M. bovis from other cultured MTC species.

  7. HANFORD DOUBLE SHELL TANK (DST) THERMAL & SEISMIC PROJECT ESTABLISHMENT OF METHODOLOGY FOR TIME DOMAIN SOIL STRUCTURE INTERACTION ANALYSIS OF HANFORD DST

    Energy Technology Data Exchange (ETDEWEB)

    MACKEY, T.C.

    2006-03-14

    the frequency domain, but frequency domain analysis is limited to systems with linear responses. The nonlinear character of the coupled SSI model and tank structural model requires that the seismic analysis be solved in the time domain. However, time domain SSI analysis is somewhat nontraditional and requires that the appropriate methodology be developed and demonstrated. Moreover, the analysis of seismically induced fluid-structure interaction between the explicitly modeled waste and the primary tank must be benchmarked against known solutions to simpler problems before being applied to the more complex analysis of the DSTs. The objective of this investigation is to establish the methodology necessary to perform the required SSI analysis of the DSTs in the time domain. Specifically, the analysis establishes the capabilities and limitations of the time domain codes ANSYS and Dytran for performing seismic SSI analysis of the DSTs. The benchmarking of the codes Dytran and ANSYS for performing seismically induced fluid-structure interaction (FSI) between the contained waste and the DST primary tank are documented in Abatt (2006) and Carpenter and Abatt (2006), respectively. The results of those two studies show that both codes have the capability to analyze the fluid-structure interaction behavior of the primary tank and contained waste. As expected, Dytran appears to have more robust capabilities for FSI analysis. The ANSYS model used in that study captures much of the FSI behavior, but does have some limitations for predicting the convective response of the waste and possibly the response of the waste in the knuckle region of the primary tank. While Dytran appears to have somewhat stronger capabilities for the analysis of the FSI behavior in the primary tank, it is more practical for the overall analysis to use ANSYS. Thus, Dytran served the purpose of helping to identify limitations in the ANSYS FSI analysis so that those limitations can be addressed in the structural

  8. Chronic breast abscess due to Mycobacterium fortuitum: a case report

    Directory of Open Access Journals (Sweden)

    MacNeill Fiona A

    2011-05-01

    Full Text Available Abstract Introduction Mycobacterium fortuitum is a rapidly growing group of nontuberculous mycobacteria more common in patients with genetic or acquired causes of immune deficiency. There have been few published reports of Mycobacterium fortuitum associated with breast infections mainly associated with breast implant and reconstructive surgery. Case presentation We report a case of a 51-year-old Caucasian woman who presented to our one-stop breast clinic with a two-week history of left breast swelling and tenderness. Following triple assessment and subsequent incision and drainage of a breast abscess, the patient was diagnosed with Mycobacterium fortuitum and treated with antibiotic therapy and surgical debridement. Conclusion This is a rare case of a spontaneous breast abscess secondary to Mycobacterium fortuitum infection. Recommended treatment is long-term antibacterial therapy and surgical debridement for extensive infection or when implants are involved.

  9. Complete Genome Sequence of Mycobacterium xenopi Type Strain RIVM700367

    KAUST Repository

    Abdallah, A. M.

    2012-05-24

    Mycobacterium xenopi is a slow-growing, thermophilic, water-related Mycobacterium species. Like other nontuberculous mycobacteria, M. xenopi more commonly infects humans with altered immune function, such as chronic obstructive pulmonary disease patients. It is considered clinically relevant in a significant proportion of the patients from whom it is isolated. We report here the whole genome sequence of M. xenopi type strain RIVM700367.

  10. A Mutant of Mycobacterium smegmatis Defective in Dipeptide Transport

    OpenAIRE

    Bhatt, Achal; Green, Renee; Coles, Roswell; Condon, Michael; Connell, Nancy D.

    1998-01-01

    A mutant of Mycobacterium smegmatis unable to use the dipeptide carnosine (β-alanyl-l-histidine) as a sole carbon or nitrogen source was isolated. Carnosinase activity and the ability to grow on β-Ala and/or l-His were similar in the mutant and the wild type. However, the mutant showed significant impairment in the uptake of carnosine. This study is the first description of a peptide utilization mutant of a mycobacterium.

  11. Chlorhexidine decontamination of sputum for culturing Mycobacterium tuberculosis

    OpenAIRE

    Asmar, Shady; Drancourt, Michel

    2015-01-01

    Background: Culture of Mycobacterium tuberculosis is the gold standard method for the laboratory diagnosis of pulmonary tuberculosis, after effective decontamination. Results: We evaluated squalamine and chlorhexidine to decontaminate sputum specimens for the culture of mycobacteria. Eight sputum specimens were artificially infected with 105 colony-forming units (cfu)/mL Mycobacterium tuberculosis and Staphylococcus aureus, Pseudomonas aeruginosa and Candida albicans as contaminants. In the s...

  12. Antibacterial Activity of Medicinal Aqueous Plant Extracts against Mycobacterium tuberculosis

    OpenAIRE

    Muna Mohammed Buzayan; Fauzia Rajab El-Garbulli

    2012-01-01

    Tuberculosis (TB) remains a serious health problem in many regions of the world, and the development of resistance to antibiotics by this microbe created the need for new drugs to replace those which have lost effectiveness. This study assesses the medicinal anti-Mycobacterium tuberculosis properties of natural products obtained from plants collected from Eastern Libya. In this study aqueous extracts of nine different plants were assayed for their Mycobacterium tuberculosis inhibitory activit...

  13. Establishing ISO 14001 Environmental Management System Analysis in a Government%创建机关ISO 14001环境管理体系探析

    Institute of Scientific and Technical Information of China (English)

    辛志伟; 顾培亮

    2001-01-01

    This paper give deep analysis of environmental elements identified which is the core in the process of establishing ISO 14001 environmental management system of Tianjin Environmental Protection Bureau as a machinery of government. General situation, characteristic, meaning of establishing of the system be introduced and analyzed. This provide a model for environmental protection bureau as a machinery of government of countrywide which estsblishing ISO 14001 environmental management system is needed.%围绕天津市环境保护局创建机关ISO 14001环境管理体系过程中的环境因素识别这一核心问题,进行了深入研究和探讨。并对体系的概况、特点和创建体系的意义等进行了概述和分析。为全国环保机关建立ISO 14001环境管理体系提供了借鉴。

  14. Establishment of turbidity forecasting model and early-warning system for source water turbidity management using back-propagation artificial neural network algorithm and probability analysis.

    Science.gov (United States)

    Yang, Tsung-Ming; Fan, Shu-Kai; Fan, Chihhao; Hsu, Nien-Sheng

    2014-08-01

    The purpose of this study is to establish a turbidity forecasting model as well as an early-warning system for turbidity management using rainfall records as the input variables. The Taipei Water Source Domain was employed as the study area, and ANOVA analysis showed that the accumulative rainfall records of 1-day Ping-lin, 2-day Ping-lin, 2-day Fei-tsui, 2-day Shi-san-gu, 2-day Tai-pin and 2-day Tong-hou were the six most significant parameters for downstream turbidity development. The artificial neural network model was developed and proven capable of predicting the turbidity concentration in the investigated catchment downstream area. The observed and model-calculated turbidity data were applied to developing the turbidity early-warning system. Using a previously determined turbidity as the threshold, the rainfall criterion, above which the downstream turbidity would possibly exceed this respective threshold turbidity, for the investigated rain gauge stations was determined. An exemplary illustration demonstrated the effectiveness of the proposed turbidity early-warning system as a precautionary alarm of possible significant increase of downstream turbidity. This study is the first report of the establishment of the turbidity early-warning system. Hopefully, this system can be applied to source water turbidity forecasting during storm events and provide a useful reference for subsequent adjustment of drinking water treatment operation. PMID:24691737

  15. The first case of cutaneous infection with Mycobacterium parascrofulaceum

    Directory of Open Access Journals (Sweden)

    Zong WK

    2012-08-01

    Full Text Available Wenkai Zong,* Xiaodong Zhang,* Hongsheng Wang, Xiu Lian Xu, Qiuling Wang, Weiwei Tian, Ya LI Jin, Qinxue Wu, Meiyu Tang Institute of Dermatology, Chinese Academy of Medical Sciences and Peking Union Medical College, National Center for STD and Leprosy Control, Chinese Center for Disease Control and Prevention, Nanjing, People’s Republic of China*These authors contributed equally to this workAbstract: The authors present the first, to the best of their knowledge, reported case of cutaneous infection caused by Mycobacterium parascrofulaceum. A 42-year-old woman presented with asymptomatic reddish papules, nodules, plaques, and patches on the right side of her face and on her forehead that had persisted for 5 years, with the lesions gradually increasing in size over that time. No previous intervening medical treatment had been applied. No history or evidence of immunosuppression was found. A skin biopsy was performed for routine histological examination. Samples of lesioned skin were inoculated on Löwenstein–Jensen medium to determine the presence of acid-fast bacilli. Ziehl–Neelsen staining was used to confirm the presence of the organism. In vitro drug susceptibility testing was conducted using the microtiter plate method. Mycobacterium was identified by polymerase chain reaction–restriction fragment length polymorphism analysis and sequencing of the hsp65 and 16S rDNA genes. Cultures for aerobic and anaerobic bacteria, as well as fungus, were also conducted. Routine histopathology revealed granulomatous changes without caseation. Ziehl–Neelsen staining showed that the organisms in both the lesions and the cultures were acid-fast bacilli. The cultured colonies were grown in Löwenstein–Jensen medium and incubated at two different temperatures (32°C and 37°C for 2–3 weeks, developing pigmentation both in the dark and in the light. In vitro drug susceptibility tests showed that the organism was sensitive to clarithromycin and

  16. Rapid differentiation of Mycobacterium xenopi from mycobacteria of the Mycobacterium avium-intracellulare complex by pyrolysis mass spectrometry.

    OpenAIRE

    Sisson, P. R.; Freeman, R.; Magee, J G; Lightfoot, N F

    1992-01-01

    Thirty four cultures of slow growing, Tween-80 negative mycobacteria were analysed by pyrolysis mass spectrometry. The results showed that pyrolysis mass spectrometry could positively distinguish strains of Mycobacterium xenopi from those of the Mycobacterium avium-intracellulare (MAI) complex. Pyrolysis mass spectrometry may be a useful technique for the rapid characterisation of non-tuberculous mycobacteria in such clinical settings as their isolation from immunocompromised patients-for exa...

  17. Septic arthritis caused by Mycobacterium fortuitum and Mycobacterium abscessus in a prosthetic knee joint: case report and review of literature.

    Science.gov (United States)

    Wang, Shu-Xiang; Yang, Chang-Jen; Chen, Yu-Chuan; Lay, Chorng-Jang; Tsai, Chen-Chi

    2011-01-01

    Nontuberculous mycobacterium (NTM) is an infrequent cause of prosthetic knee joint infections. Simultaneous infection with different NTM species in a prosthetic knee joint has not been previously reported. A case of prosthetic knee joint infection caused by Mycobacterium abscessus and M. fortuitum is described in this report. The patient was successfully treated with adequate antibiotics and surgery. The clinical features of sixteen previously reported cases of prosthetic knee joint infection caused by NTM are reviewed.

  18. Evaluation of the Abbott LCx Mycobacterium tuberculosis Assay for Direct Detection of Mycobacterium tuberculosis Complex in Human Samples

    OpenAIRE

    Garrino, M. G.; Glupczynski, Gerald; Degraux, J; Nizet, H; Delmée, Michel

    1999-01-01

    Seven hundred thirty-seven clinical samples from 460 patients were processed for direct detection of Mycobacterium tuberculosis complex by a semiautomated ligase chain reaction commercial assay, the LCx Mycobacterium tuberculosis Assay (LCx assay) from Abbott Laboratories. Results were compared to those of direct microscopy and standard microbiological culture. Of 26 patients (5.7%) with a culture positive for M. tuberculosis, 22 (84.6%) were found positive by the LCx assay. The sensitivity o...

  19. Case report of fatal Mycobacterium tilburgii infection.

    Science.gov (United States)

    Akpinar, Timur; Bakkaloglu, Oguz K; Ince, Burak; Tufan, Fatih; Kose, Murat; Poda, Mehves; Tascioglu, Didem; Koksalan, O Kaya; Saka, Bulent; Erten, Nilgun; Buyukbabani, Nesimi; Kilicaslan, Zeki; Tascioglu, Cemil

    2015-07-01

    There are few reports concerning Mycobacterium tilburgii infection in humans because this bacterium is non-cultivatable. Herein, using new molecular techniques, we report the case of an immunocompromised patient with fatal disseminated lymphadenitis that was caused by M. tilburgii.26 years old Caucasian HIV negative female patient presented with abdominal pain. Her clinical assessment revealed disseminated lymphadenitis, that was acid fast bacilli positive. Further molecular evaluation showed the causative agent as M. tilburgii. Despite anti mycobacterial therapy and careful management of intervening complications patient died because of an intraabdominal sepsis. This is the first fatal M. tilburgii infection in the literature. This case points the importance of careful management of patient's immune status and intervening infections besides implementation of effective drug treatment.

  20. The zoonotic importance of Mycobacterium bovis.

    Science.gov (United States)

    Moda, G; Daborn, C J; Grange, J M; Cosivi, O

    1996-04-01

    The zoonotic importance of Mycobacterium bovis has been the subject of renewed interest in the wake of the increasing incidence of tuberculosis in the human population. This paper considers some of the conditions under which transmission of M. bovis from animals to humans occurs and reviews current information on the global distribution of the disease. The paper highlights the particular threat posed by this zoonotic disease in developing countries and lists the veterinary and human public health measures that need to be adopted if the disease is to be contained. The association of tuberculosis with malnutrition and poverty has long been recognized and the need to address these basic issues are as crucial as specific measures against the disease itself.

  1. Evaluation of DNA microarray for detection of rifampin and isoniazid resistance in Mycobacterium tuberculosis isolates

    Institute of Scientific and Technical Information of China (English)

    王峰

    2013-01-01

    Objective To evaluate the performance of DNA microarray for rapid detection resistance to rifampin and isoniazid in Mycobacterium tuberculosis clinical isolates and identify suitable target sites for molecular genetic test. Methods Twenty-four clinical Mycobacterium

  2. 创建天津国际卫生机场的SWOT分析%SWOT analysis on the establishment of Tianjin international sanitary airport

    Institute of Scientific and Technical Information of China (English)

    赵增强; 李力军; 徐惠诚; 李瑞; 牛强

    2015-01-01

    目的:分析天津机场创建国际卫生机场的前景。方法运用SWOT分析法全面分析天津创建国际卫生机场面临的优势、劣势、机遇及风险。结果天津机场基础设施较为完善,检验检疫机构技术储备充分,基本满足创建标准要求。结论创卫工作是未来发展大势所趋,口岸检验检疫机构应积极争取地方政府支持,努力调动创建主体积极性。%Objective This paper analyzes the prospects for creating Tianjin international sanitary airport. Methods With SWOT analysis method,this paper makes a comprehensive analysis of the prospects of Tianjin international sanitary airport. Results With the sufficient technical support from CIQ and the newly-built and sound infrastructure, Tianjin airport has basically meet the requirement of international sanitary airport. Conclusion Establishing inter-national airport is a tendency in the future, inspection and quarantine agencies should actively seek the support of local government and try to mobilize the enthusiasm of airport operator.

  3. Detonation nanodiamonds for rapid detection of clinical isolates of Mycobacterium tuberculosis complex in broth culture media.

    Science.gov (United States)

    Soo, Po-Chi; Kung, Ching-Jen; Horng, Yu-Tze; Chang, Kai-Chih; Lee, Jen-Jyh; Peng, Wen-Ping

    2012-09-18

    Routinely used molecular diagnostic methods for mycobacterium identification are expensive and time-consuming. To tackle this problem, we develop a method to streamline identification of Mycobacterium tuberculosis complex (MTBC) in broth culture media by using detonation nanodiamonds (DNDs) as a platform to effectively capture the antigen secreted by MTBC which is cultured in BACTEC MGIT 960, followed by the analysis of matrix-assisted laser desorption/ionization mass spectrometry (MALDI-TOF MS). The 5 nm DNDs can capture the MTBC secretory antigen without albumin interference. With on diamond digestion, we confirm the DND captured antigen is cell filtrate protein 10 (CFP-10) because its Mascot analysis shows a score of 68. The dot blotting method further verifies a positive reaction with anti-CFP-10, indicating that CFP-10 is secreted in the medium of mycobacterium growth indicator tube (MGIT) and captured by DNDs. The minimal CFP-10 protein detection limit was 0.09 μg/mL. Furthermore, our approach can avoid the false-positive identification of MTBC by immunological methods due to cross-reactivity. Five hundred consecutive clinical specimens subjected to routine mycobacteria identification in hospital were used in this study, and the sensitivity of our method is 100% and the specificity is 98%. The analysis of each MTBC sample from culture solution can be finished within 1 h and thus shortens the turnaround time of MTBC identification of gold standard culture methods. In sum, DND MALDI-TOF MS for the detection of MTBC is rapid, specific, safe, reliable, and inexpensive. PMID:22905748

  4. Mycobacterium tuberculosis septum site determining protein, Ssd encoded by rv3660c, promotes filamentation and elicits an alternative metabolic and dormancy stress response

    Directory of Open Access Journals (Sweden)

    Crew Rebecca

    2011-04-01

    Full Text Available Abstract Background Proteins that are involved in regulation of cell division and cell cycle progression remain undefined in Mycobacterium tuberculosis. In addition, there is a growing appreciation that regulation of cell replication at the point of division is important in establishing a non-replicating persistent state. Accordingly, the objective of this study was to use a systematic approach consisting of consensus-modeling bioinformatics, ultrastructural analysis, and transcriptional mapping to identify septum regulatory proteins that participate in adaptive metabolic responses in M. tuberculosis. Results Septum site determining protein (Ssd, encoded by rv3660c was discovered to be an ortholog of septum site regulating proteins in actinobacteria by bioinformatics analysis. Increased expression of ssd in M. smegmatis and M. tuberculosis inhibited septum formation resulting in elongated cells devoid of septa. Transcriptional mapping in M. tuberculosis showed that increased ssd expression elicited a unique response including the dormancy regulon and alternative sigma factors that are thought to play a role in adaptive metabolism. Disruption of rv3660c by transposon insertion negated the unique transcriptional response and led to a reduced bacterial length. Conclusions This study establishes the first connection between a septum regulatory protein and induction of alternative metabolism consisting of alternative sigma factors and the dormancy regulon that is associated with establishing a non-replicating persistent intracellular lifestyle. The identification of a regulatory component involved in cell cycle regulation linked to the dormancy response, whether directly or indirectly, provides a foundation for additional studies and furthers our understanding of the complex mechanisms involved in establishing a non-replicating state and resumption of growth.

  5. A robust SNP barcode for typing Mycobacterium tuberculosis complex strains

    KAUST Repository

    Coll, Francesc

    2014-09-01

    Strain-specific genomic diversity in the Mycobacterium tuberculosis complex (MTBC) is an important factor in pathogenesis that may affect virulence, transmissibility, host response and emergence of drug resistance. Several systems have been proposed to classify MTBC strains into distinct lineages and families. Here, we investigate single-nucleotide polymorphisms (SNPs) as robust (stable) markers of genetic variation for phylogenetic analysis. We identify ∼92k SNP across a global collection of 1,601 genomes. The SNP-based phylogeny is consistent with the gold-standard regions of difference (RD) classification system. Of the ∼7k strain-specific SNPs identified, 62 markers are proposed to discriminate known circulating strains. This SNP-based barcode is the first to cover all main lineages, and classifies a greater number of sublineages than current alternatives. It may be used to classify clinical isolates to evaluate tools to control the disease, including therapeutics and vaccines whose effectiveness may vary by strain type. © 2014 Macmillan Publishers Limited.

  6. Detecting robust time-delayed regulation in Mycobacterium tuberculosis

    Directory of Open Access Journals (Sweden)

    Rajapakse Jagath C

    2009-12-01

    Full Text Available Abstract Background Time delays are often found in gene regulation though most techniques of building gene regulatory networks are not capable of capturing such phenomena. Here we look at the delays in the DNA repair system of Mycobacterium tuberculosis which is unusually slow in the bacteria. We propose a method based on a skip-chain model to study this phenomena in gene networks. The Viterbi paths of the underlying Markov chains find the most likely regulatory interactions among genes, taking care of very long delays. Using the derived networks, we discuss the delayed regulations and robustness of the DNA damage seen in the bacterium. Results We evaluated our method on time-course gene expressions after DNA damage with Mitocyin C. Several time-delayed interactions were observed with our analysis. The presence of hubs in the networks indicates that a small number of transcriptional factors regulate the rest of the system. We demonstrate the use of priors to overcome over-fitting problem in the generation of networks. We compare our results with the gene networks derived with dynamic Bayesian networks (DBN. Conclusion Different transcription networks are active at different stages, and constant feedback and regulation is maintained throughout the activities of a biological pathway. Skip-chain models are capable of capturing, long distant and the time-delayed regulations. Use of a Dirichlet prior over parameters and Gibbs prior over structure can greatly reduce the over-fitting in the new model.

  7. Isolation of Mycobacterium fortuitum from fish tanks in Alborz, Iran.

    Directory of Open Access Journals (Sweden)

    Shirin Akbari

    2014-08-01

    Full Text Available Fish mycobacteriosis is caused by the non-tuberculous mycobacteria. Infected fish are normally the primary source of infection, although non-tuberculous Mycobacteria can be found in the environment. The present study was designed to investigate the few recently found suspected cases of mycobacteriosis in Iranian ornamental fish tanks.Pathological specimens including granolumas from autopsied fish were used to inoculate Lowenstein-Jensen medium. Genomic material was extracted from all acid-fast positive cultures. The mycobacterial identity of bacterial isolates was authenticated using a PCR assessment targeting a 543 bp-long stretch of 16Sr RNA gene. Further more, a PCR assessment targeting a 294 bp-long stretch of heat shock protein hsp65 was performed and the amplicons were sequenced to identify the isolates.Characteristic mycobacterial bacilli were identified both in light and fluorescent microscopy of bacterial culture from all the suspected specimens. PCR-amplification of DNA templates from all isolates successfully resulted in production of the expected products. Existence of Mycobacterium fortuitum was confirmed by comparison analysis of nucleotide sequencing at hsp65 gene.The present work clearly shows mycobacteria are important in pathology of ornamental fish diseases. People who are keeping fish as pet in their homes should be cantioned about the bacterial contamination risks arise from close contact with exotic ornamental species of fish.

  8. Comparative Proteomic Profiling of Mycobacterium bovis and BCG Vaccine Strains

    KAUST Repository

    Gao, Ge

    2013-09-01

    BCG is the only licensed human vaccine currently available against TB. Derived from a virulent strain of M. bovis, the vaccine was thought to have struck a balance between reduced virulence and preserved immunogenicity. Nowadays, BCG vaccine strains used in different countries and vaccination programs show clear variations in their genomes and immune protective properties. The aim of this study was to characterize the proteomic profile on Mycobacterium bovis and five BCG strains Pasteur, Tokyo, Danish, Phipps and Birkhaug by Tandem Mass Tag® (TMT®)-labeling quantitative proteomic approach. In total, 420 proteins were identified and 377 of them were quantitated for their relative abundance. We reported the number and relationship of differential expressed proteins in BCG strains compared to M. bovis and investigated their functions by bioinformatics analysis. Several interesting up-regulated and down-regulated protein targets were found. The identified proteins and their quantitative expression profiles provide a basis for further understanding of the cellular biology of M. bovis and BCG vaccine strains, and hopefully would assist in the design of better anti-TB vaccine and drugs.

  9. Evaluation of the mutations detection of rpoB gene in Mycobacterium tuberculosis clinical isolates by probe melting analysis based real-time PCR%荧光PCR探针熔解曲线法检测结核分枝杆菌耐利福平突变研究

    Institute of Scientific and Technical Information of China (English)

    马艳艳; 李辉; 赵东阳; 李静; 李庆阁

    2011-01-01

    Objective To evaluate the analysis abilities and valuation of the melting analysis based real-time PCR for detecting the mutations of ropB gene in mycobacterium tuberculosis from clinical samples at frontier port. Methods The probe melting analysis assay was adopted for the sensitivity and specificity in detecting the isolates from clinical samples against the standard of drug susceptibility testing (DST). Results A total of 347 mycobacterium tuberculosis isolates were detected by two methods, in which, 271 rifampin sensitivity isolates and 76 rifampin resistant isolates were gained by DST. Meanwhile, 269 rifampin sensitivity isolates and 78 rifampin resistant isolates were gained by the probe melting analysis assay, the sensitivity and specificity of melting analysis assay was 93.42% and 97.42%, respectively. The concordance of the two methods was 96.54%. Conclusion The probe melting analysis assay own the characteristics of short-time, higher sensitivity and specificity for detecting rifampin resistance in tuberculosis using clinical sample and is available to use for rapid screening multidrug drug resistant tuberculosis at frontier ports.%目的 对荧光PCR熔解曲线法检测结核分枝杆菌耐利福平突变方法进行临床研究,评价检测能力及在国境口岸的应用价值.方法 应用荧光PCR熔解曲线法检测结核病人临床分离结核分枝杆菌,以传统药物敏感性试验为标准,获得该方法的灵敏性、特异性.结果 对347例结核病人临床分离培养样本,传统药物敏感性试验检出271例利福平敏感标本,76例利福平耐药标本.荧光PCR熔解曲线法检出269例利福平敏感标本,78例利福平耐药标本,灵敏性为93.42%,特异性为97.42%,符合率为96.54%.结论 荧光PCR熔解曲线法检测速度快速、灵敏度高、特异性强,可用于结核分枝杆菌利福平耐药突变的快速检测,适于国境口岸对耐多药结核病的快速筛查.

  10. 78 FR 36698 - Microbiology Devices; Reclassification of Nucleic Acid-Based Systems for Mycobacterium tuberculosis

    Science.gov (United States)

    2013-06-19

    ... Nucleic Acid-Based Systems for Mycobacterium tuberculosis Complex in Respiratory Specimens AGENCY: Food... Mycobacterium tuberculosis complex in respiratory specimens from class III (premarket approval) into class II... Mycobacterium tuberculosis Complex in Respiratory Specimens'' are necessary, in addition to general controls,...

  11. 77 FR 16126 - Microbiology Devices; Reclassification of Nucleic Acid-Based Systems for Mycobacterium tuberculosis

    Science.gov (United States)

    2012-03-19

    ...-Based Systems for Mycobacterium tuberculosis Complex AGENCY: Food and Drug Administration, HHS. ACTION...-based in vitro diagnostic devices for the detection of Mycobacterium tuberculosis complex in respiratory... Acid-Based In Vitro Diagnostic Devices for the Detection of Mycobacterium tuberculosis Complex...

  12. Purification, crystallization and preliminary X-ray crystallographic studies of Rv3705c from Mycobacterium tuberculosis

    Energy Technology Data Exchange (ETDEWEB)

    Lu, Feifei [East China University of Science and Technology, 130 Meilong Road, Shanghai 200237, People’s Republic of (China); Gao, Feng [Institute of Biophysics, Chinese Academy of Sciences, Beijing 100101, People’s Republic of (China); Li, Honglin [East China University of Science and Technology, 130 Meilong Road, Shanghai 200237, People’s Republic of (China); Gong, Weimin [Institute of Biophysics, Chinese Academy of Sciences, Beijing 100101, People’s Republic of (China); Zhou, Lin, E-mail: gdtb-bg@vip.163.com [Center for Tuberculosis Control of Guangdong Province, Guangzhou, People’s Republic of (China); Bi, Lijun, E-mail: gdtb-bg@vip.163.com [East China University of Science and Technology, 130 Meilong Road, Shanghai 200237, People’s Republic of (China)

    2014-07-23

    The cloning, expression, purification, crystallization and preliminary X-ray diffraction analysis of Rv3705c from M. tuberculosis are described. The conserved protein Rv3705c from Mycobacterium tuberculosis has been cloned, expressed, purified and crystallized by the sitting-drop vapour-diffusion method using PEG 3350 as a precipitant. The Rv3705c crystals exhibited space group P6{sub 1}22 or P6{sub 5}22, with unit-cell parameters a = b = 198.0, c = 364.1 Å, α = β = 90, γ = 120°, and diffracted to a resolution of 3.3 Å.

  13. Enhancement of beta-sitosterol transformation in Mycobacterium vaccae with increased cell wall permeability.

    Science.gov (United States)

    Korycka-Machała, M; Rumijowska-Galewicz, A; Lisowska, K; Ziolkowskit, A; Sedlacze, L

    2001-01-01

    Mycobacterium vaccae exposed to compounds which are known to disorganise the cell wall composition and architecture (protamine, glycine) showed increased specific activity in beta-sitosterol biotransformation to androstene derivatives, intennediates in the production of most medical steroids. GC/MS analysis of free lipid fatty acids revealed higher content of unsaturated compounds, mainly C16:1 and C18:1 in protamine- and glycine-treated cells than that in control cells, which seems to change the permeability features of the cell wall barrier, facilitating hydrophobic beta-sitosterol diffusion.

  14. Substantial molecular evolution and mutation rates in prolonged latent Mycobacterium tuberculosis infection in humans

    DEFF Research Database (Denmark)

    Lillebaek, Troels; Norman, Anders; Rasmussen, Erik Michael;

    2016-01-01

    confirmed cases, and used whole genome sequencing to investigate the mutational processes that occur over decades in latent Mtb. We found an estimated mutation rate between 0.2 and 0.3 over 33 years, suggesting that latent Mtb accumulates mutations at rates similar to observations from cases of active......The genome of Mycobacterium tuberculosis (Mtb) of latently infected individuals may hold the key to understanding the processes that lead to reactivation and progression to clinical disease. We report here analysis of pairs of Mtb isolates from putative prolonged latent TB cases. We identified two...

  15. Drug and Multidrug Resistance among Mycobacterium leprae Isolates from Brazilian Relapsed Leprosy Patients

    OpenAIRE

    Rocha, Adalgiza da Silva; Cunha, Maria das Graças; Diniz, Lucia Martins; Salgado, Claudio; Aires, Maria Araci P.; Nery, José Augusto; Gallo, Eugênia Novisck; Miranda, Alice; Magnanini, Monica M. F.; Matsuoka, Masanori; Sarno, Euzenir Nunes; Suffys, Philip Noel; Maria Leide W. de Oliveira

    2012-01-01

    Skin biopsy samples from 145 relapse leprosy cases and from five different regions in Brazil were submitted for sequence analysis of part of the genes associated with Mycobacterium leprae drug resistance. Single nucleotide polymorphisms (SNPs) in these genes were observed in M. leprae from 4 out of 92 cases with positive amplification (4.3%) and included a case with a mutation in rpoB only, another sample with SNPs in both folP1 and rpoB, and two cases showing mutations in folP1, rpoB, and gy...

  16. Trypanosoma Cruzi Cyp51 Inhibitor Derived from a Mycobacterium Tuberculosis Screen Hit

    Energy Technology Data Exchange (ETDEWEB)

    Chen, Chiung-Kuang; Doyle, Patricia S.; Yermalitskaya, Liudmila V.; Mackey, Zachary B.; Ang, Kenny K.H.; McKerrow, James H.; Podust, Larissa M.; (Vanderbilt); (UCSF)

    2009-02-18

    The two front-line drugs for chronic Trypanosoma cruzi infections are limited by adverse side-effects and declining efficacy. One potential new target for Chagas disease chemotherapy is sterol 14{alpha}-demethylase (CYP51), a cytochrome P450 enzyme involved in biosynthesis of membrane sterols. In a screening effort targeting Mycobacterium tuberculosis CYP51 (CYP51{sub Mt}), we previously identified the N-[4-pyridyl]-formamide moiety as a building block capable of delivering a variety of chemotypes into the CYP51 active site. In that work, the binding modes of several second generation compounds carrying this scaffold were determined by high-resolution co-crystal structures with CYP51{sub Mt}. Subsequent assays against the CYP51 orthologue in T. cruzi, CYP51{sub Tc}, demonstrated that two of the compounds tested in the earlier effort bound tightly to this enzyme. Both were tested in vitro for inhibitory effects against T. cruzi and the related protozoan parasite Trypanosoma brucei, the causative agent of African sleeping sickness. One of the compounds had potent, selective anti-T. cruzi activity in infected mouse macrophages. Cure of treated host cells was confirmed by prolonged incubation in the absence of the inhibiting compound. Discrimination between T. cruzi and T. brucei CYP51 by the inhibitor was largely based on the variability (phenylalanine versus isoleucine) of a single residue at a critical position in the active site. CYP51{sub Mt}-based crystal structure analysis revealed that the functional groups of the two tightly bound compounds are likely to occupy different spaces in the CYP51 active site, suggesting the possibility of combining the beneficial features of both inhibitors in a third generation of compounds to achieve more potent and selective inhibition of CYP51{sub Tc}. Enzyme sterol 14{alpha}-demethylase (CYP51) is a well-established target for anti-fungal therapy and is a prospective target for Chagas disease therapy. We previously identified a

  17. Mutations in katG gene sequences in isoniazid-resistant clinical isolates of Mycobacterium tuberculosis are rare.

    OpenAIRE

    Pretorius, G.S.; van Helden, P. D.; Sirgel, F; Eisenach, K D; Victor, T C

    1995-01-01

    In this study, a battery of oligonucleotides was directed toward the katG gene and PCR-single-stranded conformation polymorphism (SSCP) analysis was used to search for katG gene deviations in clinical isolates of Mycobacterium tuberculosis from different geographical regions. Since a complete deletion of the katG gene was not found, it is suggested that deletion is not a major mechanism of isoniazid (isonicotinic acid hydrazide; INH) resistance in these isolates. However, 7 of 39 isolates (4 ...

  18. Pathogen-Specific Epitopes as Epidemiological Tools for Defining the Magnitude of Mycobacterium leprae Transmission in Areas Endemic for Leprosy

    OpenAIRE

    Martins, Marcia V. S. B.; Guimarães, Marjorie M. da S.; Spencer, John S.; Mariana A V B Hacker; Luciana S Costa; Fernanda M Carvalho; Annemieke Geluk; van der Ploeg-van Schip, Jolien J.; Pontes, Maria A. A.; Heitor S Gonçalves; de Morais, Janvier P.; Bandeira, Tereza J.P.G.; Pessolani, Maria C. V.; Brennan, Patrick J.; Pereira, Geraldo M. B.

    2012-01-01

    During recent years, comparative genomic analysis has allowed the identification of Mycobacterium leprae-specific genes with potential application for the diagnosis of leprosy. In a previous study, 58 synthetic peptides derived from these sequences were tested for their ability to induce production of IFN-γ in PBMC from endemic controls (EC) with unknown exposure to M. leprae, household contacts of leprosy patients and patients, indicating the potential of these synthetic peptides for the dia...

  19. Simultaneous detection and strain differentiation of Mycobacterium tuberculosis complex in paraffin wax embedded tissues and in stained microscopic preparations.

    OpenAIRE

    Zanden, A.G. van der; Hoentjen, A H; Heilmann, F G; Weltevreden, E F; Schouls, L. M.; van Embden, J D

    1998-01-01

    AIMS: To detect and differentiate Mycobacterium tuberculosis simultaneously by polymerase chain reaction (PCR) in clinical samples prepared for histopathological analysis and for microscopic detection of acid fast bacteria. METHODS: Paraffin wax embedded tissue samples and Ziehl-Neelsen (ZN) and auramine stained microscopic preparations from culture positive tuberculosis patients were subjected to DNA extraction and amplification by PCR. PCR was performed with primers specific for direct repe...

  20. Syndrome of selective IgM deficiency with severe T cell deficiency associated with disseminated cutaneous mycobacterium avium intracellulaire infection

    OpenAIRE

    Gharib, Asal; Louis, Ankmalika Gupta; Agrawal, Sudhanshu; Gupta, Sudhir

    2015-01-01

    Cutaneous non-disseminated, non-tuberculous mycobacterial infections have been reported in both immunocompetent and immunocompromised subjects. Systemic Mycobacterium avium intracellulaire (MAI) have been reported in non-HIV patients with Idiopathic CD4 lymphocytopenia. We report a comprehensive immunological analysis in syndrome of selective IgM deficiency and T lymphocytopenia (both CD4+ and CD8+) with disseminated cutaneous MAI infection. Naïve (TN) and Central memory (TCM) subsets of both...

  1. Analysis of Mycobacterium tuberculosis DNA detection results of 3860 different samples%3860例不同标本结核分枝杆菌DNA检测结果分析

    Institute of Scientific and Technical Information of China (English)

    李锐成; 刘昕阳; 邹菊贤; 闫琳; 杨文青; 张惠中

    2013-01-01

    目的:了解实时荧光定量PCR检测TB-DNA的临床意义及不同标本类型TB-DNA检出率的差异.方法:采用实时荧光定量PCR法对患者的血清、尿液、痰液、全血、胸腹水、脑脊液、穿刺液、灌洗液进行结核菌DNA检测.结果:3860份标本的总阳性率为2.82%,血清标本的阳性率在所有标本类型中最低,仅为0.58%,穿刺液和尿液的阳性率较高,分别为10.60%和23.18%.结论:实时荧光定量TB-DNA具有快速、简便、敏感性高、特异性强的特点,适宜多种标本类型的检测,是临床上早期诊断结核菌感染的有效方法.%Objective:To investigate the clinic significance of real-time fluorescent quantitative PCR in detection of Mycobacterium tuberculosis DNA (TB-DNA),and the differential detection rate of 8 types of samples.Methods:The samples of serum,urine,sputum,whole blood,irrigating solution,puncture fluid,cerebrospinal fluid,pleural fluid and ascites were detected for TB-DNA by real-time fluorescent quantitative PCR assay.Results:Totally 3860 different samples were detected and the total positive rate was 2.82%.Serum had the lowest positive rate among the 8 types of samples,only 0.58%.Puncture fluid and urine had higher positive rate with 10.60% and 23.18% respectively.Conclusion:Real-time fluorescent quantitative PCR is fast,simple,high sensitive and specific,and suitable for detection of different samples,which is one of the effective methods for detection of TB-DNA in early diagnosis of tuberculosis.

  2. Virulence-related Mycobacterium avium subsp hominissuis MAV_2928 gene is associated with vacuole remodeling in macrophages

    Directory of Open Access Journals (Sweden)

    Vogt Steven

    2010-04-01

    Full Text Available Abstract Background Mycobacterium avium subsp hominissuis (previously Mycobacterium avium subsp avium is an environmental organism associated with opportunistic infections in humans. Mycobacterium hominissuis infects and replicates within mononuclear phagocytes. Previous study characterized an attenuated mutant in which the PPE gene (MAV_2928 homologous to Rv1787 was inactivated. This mutant, in contrast to the wild-type bacterium, was shown both to have impaired the ability to replicate within macrophages and to have prevented phagosome/lysosome fusion. Results MAV_2928 gene is primarily upregulated upon phagocytosis. The transcriptional profile of macrophages infected with the wild-type bacterium and the mutant were examined using DNA microarray, which showed that the two bacteria interact uniquely with mononuclear phagocytes. Based on the results, it was hypothesized that the phagosome environment and vacuole membrane of the wild-type bacterium might differ from the mutant. Wild-type bacterium phagosomes expressed a number of proteins different from those infected with the mutant. Proteins on the phagosomes were confirmed by fluorescence microscopy and Western blot. The environment in the phagosome of macrophages infected with the mutant differed from the environment of vacuoles with M. hominissuis wild-type in the concentration of zinc, manganese, calcium and potassium. Conclusion The results suggest that the MAV_2928 gene/operon might participate in the establishment of bacterial intracellular environment in macrophages.

  3. Identification of specific metabolites in culture supernatant of Mycobacterium tuberculosis using metabolomics: exploration of potential biomarkers.

    Science.gov (United States)

    Lau, Susanna K P; Lam, Ching-Wan; Curreem, Shirly O T; Lee, Kim-Chung; Lau, Candy C Y; Chow, Wang-Ngai; Ngan, Antonio H Y; To, Kelvin K W; Chan, Jasper F W; Hung, Ivan F N; Yam, Wing-Cheong; Yuen, Kwok-Yung; Woo, Patrick C Y

    2015-01-01

    Although previous studies have reported the use of metabolomics for Mycobacterium species differentiation, little is known about the potential of extracellular metabolites of Mycobacterium tuberculosis (MTB) as specific biomarkers. Using an optimized ultrahigh performance liquid chromatography-electrospray ionization-quadruple time of flight-mass spectrometry (UHPLC-ESI-Q-TOF-MS) platform, we characterized the extracellular metabolomes of culture supernatant of nine MTB strains and nine non-tuberculous Mycobacterium (NTM) strains (four M. avium complex, one M. bovis Bacillus Calmette-Guérin (BCG), one M. chelonae, one M. fortuitum and two M. kansasii). Principal component analysis readily distinguished the metabolomes between MTB and NTM. Using multivariate and univariate analysis, 24 metabolites with significantly higher levels in MTB were identified. While seven metabolites were identified by tandem mass spectrometry (MS/MS), the other 17 metabolites were unidentified by MS/MS against database matching, suggesting that they may be potentially novel compounds. One metabolite was identified as dexpanthenol, the alcohol analog of pantothenic acid (vitamin B5), which was not known to be produced by bacteria previously. Four metabolites were identified as 1-tuberculosinyladenosine (1-TbAd), a product of the virulence-associated enzyme Rv3378c, and three previously undescribed derivatives of 1-TbAd. Two derivatives differ from 1-TbAd by the ribose group of the nucleoside while the other likely differs by the base. The remaining two metabolites were identified as a tetrapeptide, Val-His-Glu-His, and a monoacylglycerophosphoglycerol, phosphatidylglycerol (PG) (16∶0/0∶0), respectively. Further studies on the chemical structure and biosynthetic pathway of these MTB-specific metabolites would help understand their biological functions. Studies on clinical samples from tuberculosis patients are required to explore for their potential role as diagnostic biomarkers. PMID

  4. Combining groundwater quality analysis and a numerical flow simulation for spatially establishing utilization strategies for groundwater and surface water in the Pingtung Plain

    Science.gov (United States)

    Jang, Cheng-Shin; Chen, Ching-Fang; Liang, Ching-Ping; Chen, Jui-Sheng

    2016-02-01

    Overexploitation of groundwater is a common problem in the Pingtung Plain area of Taiwan, resulting in substantial drawdown of groundwater levels as well as the occurrence of severe seawater intrusion and land subsidence. Measures need to be taken to preserve these valuable groundwater resources. This study seeks to spatially determine the most suitable locations for the use of surface water on this plain instead of extracting groundwater for drinking, irrigation, and aquaculture purposes based on information obtained by combining groundwater quality analysis and a numerical flow simulation assuming the planning of manmade lakes and reservoirs to the increase of water supply. The multivariate indicator kriging method is first used to estimate occurrence probabilities, and to rank townships as suitable or unsuitable for groundwater utilization according to water quality standards for drinking, irrigation, and aquaculture. A numerical model of groundwater flow (MODFLOW) is adopted to quantify the recovery of groundwater levels in townships after model calibration when groundwater for drinking and agricultural demands has been replaced by surface water. Finally, townships with poor groundwater quality and significant increases in groundwater levels in the Pingtung Plain are prioritized for the groundwater conservation planning based on the combined assessment of groundwater quality and quantity. The results of this study indicate that the integration of groundwater quality analysis and the numerical flow simulation is capable of establishing sound strategies for joint groundwater and surface water use. Six southeastern townships are found to be suitable locations for replacing groundwater with surface water from manmade lakes or reservoirs to meet drinking, irrigation, and aquaculture demands.

  5. Establishment of An Air Pollutant Emission Inventory and Uncertainty Analysis%大气污染物排放清单的建立及不确定性

    Institute of Scientific and Technical Information of China (English)

    薛亦峰; 闫静; 宋光武; 李金玉

    2012-01-01

    On the basis of the investigation and the study on both domestic and foreign air pollutant emission inventories,the study summarized the basic procedures for an inventory,introduced the process of the establishment of an air pollutant emission inventory,inventory management,improvement,quality assurance and control plan.As an important aspect of improving the inventory,uncertainty analysis was necessary.This study described the source of its uncertainty and quantitative or qualitative evaluation methods.%本文在调研国内外大气污染物排放清单的基础上,总结了清单编制的基本程序,介绍大气污染物排放清单的建立,包括技术路线和计算方法,以及清单的管理、改善及质量保证与控制计划。不确定性分析是完善清单的重要方面,本文阐述了清单不确定性的来源及定量、定性的评价方法。

  6. Micrococcin P1 - A bactericidal thiopeptide active against Mycobacterium tuberculosis.

    Science.gov (United States)

    Degiacomi, Giulia; Personne, Yoann; Mondésert, Guillaume; Ge, Xueliang; Mandava, Chandra Sekhar; Hartkoorn, Ruben C; Boldrin, Francesca; Goel, Pavitra; Peisker, Kristin; Benjak, Andrej; Barrio, Maria Belén; Ventura, Marcello; Brown, Amanda C; Leblanc, Véronique; Bauer, Armin; Sanyal, Suparna; Cole, Stewart T; Lagrange, Sophie; Parish, Tanya; Manganelli, Riccardo

    2016-09-01

    The lack of proper treatment for serious infectious diseases due to the emergence of multidrug resistance reinforces the need for the discovery of novel antibiotics. This is particularly true for tuberculosis (TB) for which 3.7% of new cases and 20% of previously treated cases are estimated to be caused by multi-drug resistant strains. In addition, in the case of TB, which claimed 1.5 million lives in 2014, the treatment of the least complicated, drug sensitive cases is lengthy and disagreeable. Therefore, new drugs with novel targets are urgently needed to control resistant Mycobacterium tuberculosis strains. In this manuscript we report the characterization of the thiopeptide micrococcin P1 as an anti-tubercular agent. Our biochemical experiments show that this antibiotic inhibits the elongation step of protein synthesis in mycobacteria. We have further identified micrococcin resistant mutations in the ribosomal protein L11 (RplK); the mutations were located in the proline loop at the N-terminus. Reintroduction of the mutations into a clean genetic background, confirmed that they conferred resistance, while introduction of the wild type RplK allele into resistant strains re-established sensitivity. We also identified a mutation in the 23S rRNA gene. These data, in good agreement with previous structural studies suggest that also in M. tuberculosis micrococcin P1 functions by binding to the cleft between the 23S rRNA and the L11 protein loop, thus interfering with the binding of elongation factors Tu and G (EF-Tu and EF-G) and inhibiting protein translocation. PMID:27553416

  7. Novel multiplex real-time PCR diagnostic assay for identification and differentiation of Mycobacterium tuberculosis, Mycobacterium canettii, and Mycobacterium tuberculosis complex strains.

    NARCIS (Netherlands)

    Reddington, K.; O'Grady, J.; Dorai-Raj, S.; Maher, M.; Soolingen, D. van; Barry, T.

    2011-01-01

    Tuberculosis (TB) in humans is caused by members of the Mycobacterium tuberculosis complex (MTC). Rapid detection of the MTC is necessary for the timely initiation of antibiotic treatment, while differentiation between members of the complex may be important to guide the appropriate antibiotic treat

  8. Mycobacterium avium subsp. hominissuis Infection in Swine Associated with Peat Used for Bedding

    Directory of Open Access Journals (Sweden)

    Tone Bjordal Johansen

    2014-01-01

    Full Text Available Mycobacterium avium subsp. hominissuis is an environmental bacterium causing opportunistic infections in swine, resulting in economic losses. Additionally, the zoonotic aspect of such infections is of concern. In the southeastern region of Norway in 2009 and 2010, an increase in condemnation of pig carcasses with tuberculous lesions was seen at the meat inspection. The use of peat as bedding in the herds was suspected to be a common factor, and a project examining pigs and environmental samples from the herds was initiated. Lesions detected at meat inspection in pigs originating from 15 herds were sampled. Environmental samples including peat from six of the herds and from three peat production facilities were additionally collected. Samples were analysed by culture and isolates genotyped by MLVA analysis. Mycobacterium avium subsp. hominissuis was detected in 35 out of 46 pigs, in 16 out of 20 samples of peat, and in one sample of sawdust. MLVA analysis demonstrated identical isolates from peat and pigs within the same farms. Polyclonal infection was demonstrated by analysis of multiple isolates from the same pig. To conclude, the increase in condemnation of porcine carcasses at slaughter due to mycobacteriosis seemed to be related to untreated peat used as bedding.

  9. Relatedness of Mycobacterium avium subspecies hominissuis clinical isolates of human and porcine origins assessed by MLVA.

    Science.gov (United States)

    Leão, Célia; Canto, Ana; Machado, Diana; Sanches, Ilda Santos; Couto, Isabel; Viveiros, Miguel; Inácio, João; Botelho, Ana

    2014-09-17

    Mycobacterium avium subsp. hominissuis (MAH) is an important opportunistic pathogen, infecting humans and animals, notably pigs. Several methods have been used to characterize MAH strains. RFLP and PFGE typing techniques have been used as standard methods but are technically demanding. In contrast, the analysis of VNTR loci is a simpler, affordable and highly reliable PCR-based technique, allowing a numerical and reproductive digitalization of typing data. In this study, the analysis of Mycobacterium avium tandem repeats (MATRs) loci was adapted to evaluate the genetic diversity of epidemiological unrelated MAH clinical strains of human (n=28) and porcine (n=69) origins, collected from diverse geographical regions across mainland Portugal. These MAH isolates were found to be genetically diverse and genotypes are randomly distributed across the country. Some of the human strains shared identical VNTR profiles with porcine isolates. Our study shows that the VNTR genotyping using selected MATR loci is a useful analysis technique for assessing the genetic diversity of MAH isolates from Portugal. This typing method could be successfully applied in other countries toward the implementation of a worldwide open-access database of MATR-VNTR profiles of MAH isolates, allowing a better assessment of the global epidemiology traits of this important pathogenic species.

  10. Advances in establishment and analysis of three-dimensional tumor spheroid-based functional assays for target validation and drug evaluation

    Directory of Open Access Journals (Sweden)

    Vinci Maria

    2012-03-01

    Full Text Available Abstract Background There is overwhelming evidence that in vitro three-dimensional tumor cell cultures more accurately reflect the complex in vivo microenvironment than simple two-dimensional cell monolayers, not least with respect to gene expression profiles, signaling pathway activity and drug sensitivity. However, most currently available three-dimensional techniques are time consuming and/or lack reproducibility; thus standardized and rapid protocols are urgently needed. Results To address this requirement, we have developed a versatile toolkit of reproducible three-dimensional tumor spheroid models for dynamic, automated, quantitative imaging and analysis that are compatible with routine high-throughput preclinical studies. Not only do these microplate methods measure three-dimensional tumor growth, but they have also been significantly enhanced to facilitate a range of functional assays exemplifying additional key hallmarks of cancer, namely cell motility and matrix invasion. Moreover, mutual tissue invasion and angiogenesis is accommodated by coculturing tumor spheroids with murine embryoid bodies within which angiogenic differentiation occurs. Highly malignant human tumor cells were selected to exemplify therapeutic effects of three specific molecularly-targeted agents: PI-103 (phosphatidylinositol-3-kinase (PI3K-mammalian target of rapamycin (mTOR inhibitor, 17-N-allylamino-17-demethoxygeldanamycin (17-AAG (heat shock protein 90 (HSP90 inhibitor and CCT130234 (in-house phospholipase C (PLCγ inhibitor. Fully automated analysis using a Celigo cytometer was validated for tumor spheroid growth and invasion against standard image analysis techniques, with excellent reproducibility and significantly increased throughput. In addition, we discovered key differential sensitivities to targeted agents between two-dimensional and three-dimensional cultures, and also demonstrated enhanced potency of some agents against cell migration

  11. The relative frequency of Mycobacterium tuberculosis and Mycobacterium avium infections in HIV positive patients, Ahvaz, Iran

    Institute of Scientific and Technical Information of China (English)

    Khosravi AD; Alavi SM; Hashemzade M; Abasi E; Seghatoleslami S

    2012-01-01

    Objective:To estimate the prevalence of Mycobacterium tuberculosis (M. tuberculosis) and Mycobacterium avium (M. avium) infections in HIV-positive patients suspected to have pulmonary and extrapulmonary mycobacterial co-infection using PCR technique. Methods:Totally 50 samples comprising sputum, pleural fluid and CSF taken from HIV positive patients suspected to have mycobacterial infection, were processed. The demographic information and results of acid fast staining and culture were recorded for each patient. The PCR for detecting of M. tuberculosis comprised of specific primers targeting IS6110 gene sequence. For detecting of M. avium, PCR with primers that amplifies the mig gene were used. Results:From 50 samples processed, 45 were sputum (90%), 3 pleural fluid (6%) and 2 CSF (4%). In total, 8 (16%) were culture positive, 7 had positive acid fast staining (14%) and 13 samples (26%) were positive using PCR technique. All the positive samples were sputum and belonged to patients with pulmonary infection. Of these, 9 were positive for M. tuberculosis (69.2%) and 4 were identified as M. avium (30.8%), which 2 out of 13 positive samples showed mixed infections by both mycobacteria. Conclusions:The PCR shows the highest detection rate (26%) of mycobacteria compared with culture and acid fast staining. The majority of infections were with M. tuberculosis (18%) and this shows the importance of this mycobacterial co-infection in HIV positive patients in the region of study.

  12. Differences in T-cell responses between Mycobacterium tuberculosis and Mycobacterium africanum-infected patients.

    Science.gov (United States)

    Tientcheu, Leopold D; Sutherland, Jayne S; de Jong, Bouke C; Kampmann, Beate; Jafali, James; Adetifa, Ifedayo M; Antonio, Martin; Dockrell, Hazel M; Ota, Martin O

    2014-05-01

    In The Gambia, Mycobacterium tuberculosis (Mtb) and Mycobacterium africanum (Maf) are major causes of tuberculosis (TB). Maf is more likely to cause TB in immune suppressed individuals, implying differences in virulence. Despite this, few studies have assessed the underlying immunity to the two pathogens in human. In this study, we analyzed T-cell responses from 19 Maf- and 29 Mtb-infected HIV-negative patients before and after TB chemotherapy following overnight stimulation of whole blood with TB-specific antigens. Before treatment, percentages of early secreted antigenic target-6(ESAT-6)/culture filtrate protein-10(CFP-10) and purified protein derivative-specific single-TNF-α-producing CD4(+) and CD8(+) T cells were significantly higher while single-IL-2-producing T cells were significantly lower in Maf- compared with Mtb-infected patients. Purified protein derivative-specific polyfunctional CD4(+) T cells frequencies were significantly higher before than after treatment, but there was no difference between the groups at both time points. Furthermore, the proportion of CD3(+) CD11b(+) T cells was similar in both groups pretreatment, but was significantly lower with higher TNF-α, IL-2, and IFN-γ production in Mtb- compared with that of Maf-infected patients posttreatment. Our data provide evidence of differences in T-cell responses to two mycobacterial strains with differing virulence, providing some insight into TB pathogenesis with different Mtb strains that could be prospectively explored as biomarkers for TB protection or susceptibility.

  13. Pengembangan Media Padat untuk Menumbuhkan Mycobacterium bovis (DEVELOPMENT OF SOLID MEDIUM FOR MYCOBACTERIUM BOVIS CULTIVATION

    Directory of Open Access Journals (Sweden)

    Mazdani Ulfah Daulay

    2016-01-01

    Full Text Available Mycobacterial culture provides definitive diagnosis of tuberculosis (TB, but commercially readyto-use culture media for Mycobacterium bovis are rarely available. The aims of this study were todevelop and to evaluate the ability of M. Bovis to grow in Modified Ogawa Agar (MOA in comparisonwith the available culture media, such as Löwenstein Jensen (LJ and Modified Ogawa (MO. Eachmedia were inculation with 0.1 ml suspension of 105 CFU/mL M. bovis and M. phlei in PhosphateBuffer Saline (PBS and each media was replicated in five tubes. Mycobacterium phlei grew in everymedium since day 4. M. bovis grew in media LJ and MO since day 17, but failed to grow in mediumMOA. The recovery rate of M. phlei in LJ and MOA were significantly different. The ability of MOA tocultivate M. phlei was different from LJ. Colonies of M. phlei in MOA were easier to be harvested, muchsimpler to prepare, and more feasible than medium LJ. The recovery rate of M. bovis in media LJ andMO were not significantly different, but medium MO were much simpler to prepare and more feasiblethan medium LJ. Media MOA were able to cultivate M. phlei, but proven unable to cultivate M. bovisin this research.

  14. Divergent immune responses to Mycobacterium avium subsp. paratuberculosis infection correlate with kinome responses at the site of intestinal infection.

    Science.gov (United States)

    Määttänen, Pekka; Trost, Brett; Scruten, Erin; Potter, Andrew; Kusalik, Anthony; Griebel, Philip; Napper, Scott

    2013-08-01

    Mycobacterium avium subsp. paratuberculosis is the causative agent of Johne's disease (JD) in cattle. M. avium subsp. paratuberculosis infects the gastrointestinal tract of calves, localizing and persisting primarily in the distal ileum. A high percentage of cattle exposed to M. avium subsp. paratuberculosis do not develop JD, but the mechanisms by which they resist infection are not understood. Here, we merge an established in vivo bovine intestinal segment model for M. avium subsp. paratuberculosis infection with bovine-specific peptide kinome arrays as a first step to understanding how infection influences host kinomic responses at the site of infection. Application of peptide arrays to in vivo tissue samples represents a critical and ambitious step in using this technology to understand host-pathogen interactions. Kinome analysis was performed on intestinal samples from 4 ileal segments subdivided into 10 separate compartments (6 M. avium subsp. paratuberculosis-infected compartments and 4 intra-animal controls) using bovine-specific peptide arrays. Kinome data sets clustered into two groups, suggesting unique binary responses to M. avium subsp. paratuberculosis. Similarly, two M. avium subsp. paratuberculosis-specific immune responses, characterized by different antibody, T cell proliferation, and gamma interferon (IFN-γ) responses, were also observed. Interestingly, the kinomic groupings segregated with the immune response groupings. Pathway and gene ontology analyses revealed that differences in innate immune and interleukin signaling and particular differences in the Wnt/β-catenin pathway distinguished the kinomic groupings. Collectively, kinome analysis of tissue samples offers insight into the complex cellular responses induced by M. avium subsp. paratuberculosis in the ileum and provides a novel method to understand mechanisms that alter the balance between cell-mediated and antibody responses to M. avium subsp. paratuberculosis infection. PMID

  15. Whole blood assay to access T cell-immune responses to Mycobacterium tuberculosis antigens in healthy Brazilian individuals

    Directory of Open Access Journals (Sweden)

    Paulo RZ Antas

    2004-02-01

    Full Text Available The production of interferon gamma (IFNgamma guarantees effective T cell-mediated immunity against Mycobacterium tuberculosis infection. In the present study, we simply compare the in vitro immune responses to Mycobacterium antigens in terms of IFNg production in a total of 10 healthy Brazilian volunteers. Whole blood and mononuclear cells were cultivated in parallel with PPD, Ag85B, and M. bovis hsp65, and five-days supernatants were harvested for cytokine detection by ELISA. The inter-assay result was that the overall profile of agreement in response to antigens was highly correlated (r² = 0.9266; p = 0.0102. Potential analysis is in current progress to dictate the usefulness of this method to access the immune responses also in tuberculosis patients and its contacts.

  16. Analysis on the current situation of non-tuberculosis mycobacterium in Ningbo%宁波地区非结核分枝杆菌流行状况分析

    Institute of Scientific and Technical Information of China (English)

    车洋; 于梅; 平国华

    2012-01-01

    Objective: To analyze the current situation and trend of non - tuberculosis mycobacterium in Ningbo and to provide information for diagnosis, treatment and policy on tuberculosis control. Methods: Relevant data regarding Mycobacteria culture, species identification and drug - resistance from our patients under suspicion of having pulmonary tuberculosis seen at TB control lab, was analyzed retrospectively during 2005 -2007. Results: A total number of 1917 strains of Mycobacteria were isolated and 335 strains were identified as NTM, which accounted for 17.48% of the isolated strains during 3 years. The annual isolation rates of NTM were between 13. 20% and 20.53%. When compared with 2005, the rates of isolation on NTM had increased 7. 33% in 2007. 166 strains were resistant at least to rifampin and isoniazid out of 335 NTM strains tested for drug susceptibility to isoniazid, ri-fampin, streptomycin and ethambutol. The average rate of multi - drugs resistance of these strains was 49. 55%. People at 25 years old or older, especially the 25 ~ 45 years old, were more susceptible to NTM than other age groups. Sex ratio was 2. 32:1. Contusion; The current situation and trend of NTM in Ningbo were nearly the same as National Tuberculosis Epidemiology. However, the rising tendency of rate of isolation, mainly consisted of opportunistic pathogens and the surprisingly high rate of drug - resistance to NTM both call for special attention. Studies regarding the epidemiology of NTM shoud be posed and implemented in the National TB Control Program.%目的:探讨宁波地区非结核分枝杆菌(NTM)流行状况及趋势.方法:回顾分析2005年-2007年宁波市结核病防治所参比实验室分枝杆菌耐药性监测资料.结果:1917株分枝杆菌中,非结核分枝杆菌为335株,分离率为17.48%.年分离率为13.20% ~20.53%,3年上升幅度为7.33%;对335株非结核分枝杆菌进行异烟肼(H)、利福平(R)、乙胺丁醇(E)和链霉素(S)4种抗结

  17. Mycobacterium kansasii pulmonary diseases in Korea.

    Science.gov (United States)

    Yim, Jae-Joon; Park, Young-Kil; Lew, Woo Jin; Bai, Gill-Han; Han, Sung Koo; Shim, Young-Soo

    2005-12-01

    Mycobacterium kansasii is one of the most common cause of pulmonary diseases due to nontuberculous mycobacteria. We investigated the changing in the number of isolation of M. kansasii and the clinical characteristics of M. kansasii pulmonary disease in Korea. Through searching the database of the Korean Institute of Tuberculosis, we identified the cases of isolated M. kansasii from 1992 to 2002. The number of M. kansasii isolation had increased from once in 1992 to 62 in 2002. Fifteen patients with M. kansasii pulmonary disease were identified during the period January 1997 to December 2002. Twelve patients (80%) were male and fourteen (93%) were from highly industrialized areas. The most common symptom was a cough. Seven patients (47%) had a cavitary lesion and right upper lobe was most commonly involved. Patients responded well to isoniazid and rifampicin based regimens both bacteriologically and radiographically. In conclusion, M. kansasii isolation has increased, especially in highly industrialized areas, as well as other nontuberculous mycobacteria in Korea. PMID:16361804

  18. Autophagy in Mycobacterium tuberculosis and HIV infections

    Directory of Open Access Journals (Sweden)

    Lucile eEspert

    2015-06-01

    Full Text Available Human Immunodeficiency Virus (HIV and Mycobacterium tuberculosis (M.tb are among the most lethal human pathogens worldwide, each being responsible for around 1.5 million deaths annually. Moreover, synergy between acquired immune deficiency syndrome (AIDS and tuberculosis (TB has turned HIV/M.tb co-infection into a major public health threat in developing countries. In the past decade, autophagy, a lysosomal catabolic process, has emerged as a major host immune defense mechanism against infectious agents like M.tb and HIV. Nevertheless, in some instances, autophagy machinery appears to be instrumental for HIV infection. Finally, there is mounting evidence that both pathogens deploy various countermeasures to thwart autophagy. This mini-review proposes an overview of the roles and regulations of autophagy in HIV and M.tb infections with an emphasis on microbial factors. We also discuss the role of autophagy manipulation in the context of HIV/M.tb co-infection. In future, a comprehensive understanding of autophagy interaction with these pathogens will be critical for development of autophagy-based prophylactic and therapeutic interventions for AIDS and TB.

  19. Fish tank granuloma caused by Mycobacterium marinum.

    Directory of Open Access Journals (Sweden)

    Ting-Shu Wu

    Full Text Available INTRODUCTION: Mycobacterium marinum causes skin and soft tissue, bone and joint, and rare disseminated infections. In this study, we aimed to investigate the relationship between treatment outcome and antimicrobial susceptibility patterns. A total of 27 patients with M. marinum infections were enrolled. METHODS: Data on clinical characteristics and therapeutic methods were collected and analyzed. We also determined the minimum inhibitory concentrations of 7 antibiotics against 30 isolates from these patients. RESULTS: Twenty-seven patients received antimycobacterial agents with or without surgical debridement. Eighteen patients were cured, 8 failed to respond to treatment, and one was lost to follow-up. The duration of clarithromycin (147 vs. 28; p = 0.0297, and rifampicin (201 vs. 91; p = 0.0266 treatment in the cured patients was longer than that in the others. Surgical debridement was performed in 10 out of the 18 cured patients, and in 1 of another group (p = 0.0417. All the 30 isolates were susceptible to clarithromycin, amikacin, and linezolid; 29 (96.7% were susceptible to ethambutol; 28 (93.3% were susceptible to sulfamethoxazole; and 26 (86.7% were susceptible to rifampicin. However, only 1 (3.3% isolate was susceptible to doxycycline. DISCUSSION: Early diagnosis of the infection and appropriate antimicrobial therapy with surgical debridement are the mainstays of successful treatment. Clarithromycin and rifampin are supposed to be more effective agents.

  20. Therapeutic keratectomy for Mycobacterium abscessus keratitis after LASIK.

    Science.gov (United States)

    Sun, Yi-Chen; Wang, I-Jong; Chen, Wei-Li; Hu, Fung-Rong

    2003-11-01

    We report successful treatment of a case of Mycobacterium abscessus keratitis after laser in situ keratomileusis (LASIK) with therapeutic lamellar keratectomy. A 34-year-old woman developed a 2 x 2 mm feathery infiltration within the interface inferior to the pupil margin with mild inflammation of the conjunctiva in her left eye 40 days after LASIK surgery. Bacterial culture from the infiltrates of the interface of the stromal bed revealed Mycobacterium abscessus. After combination antibiotic therapy including amikacin and ciprofoxacin was given for 6 weeks, infiltration persisted despite the development of necrosis in the flap tissue. Therapeutic lamellar keratectomy combined with flap removal was performed. No recurrence was found 1 year after the surgery. Therapeutic lamellar keratectomy with flap removal can provide an effective treatment modality for the management of post-LASIK Mycobacterium abscessus keratitis that is unresponsive to medical treatment. PMID:14724729

  1. Fatal aortic pseudoaneurysm from disseminated Mycobacterium kansasii infection: case report.

    Science.gov (United States)

    Ehsani, Laleh; Reddy, Sujan C; Mosunjac, Mario; Kraft, Colleen S; Guarner, Jeannette

    2015-03-01

    Mycobacterium kansasii is a photochromogenic, slow-growing mycobacterium species that can cause pulmonary infection in patients with predisposing lung diseases, as well as extrapulmonary or disseminated disease in immunosuppressed patients. We describe a patient with a myelodysplastic syndrome, disseminated M kansasii infection, and ruptured aortic aneurysm. He had a recent diagnosis of mycobacterium cavitary lung lesions and was transferred to our facility for possible surgical intervention of an aortic aneurysm. Few hours after admission, the patient suddenly collapsed and died despite resuscitation efforts. A complete autopsy was performed and showed ruptured ascending aortic pseudoaneurysm with hemopericardium, disseminated necrotizing and nonnecrotizing granulomas with acid-fast bacilli in the aortic wall, lungs, heart, liver, spleen, and kidneys. Further genetic studies were consistent with monocytopenia and mycobacterial infection syndrome. PMID:25537975

  2. Establishment Registration & Device Listing

    Data.gov (United States)

    U.S. Department of Health & Human Services — This searchable database contains establishments (engaged in the manufacture, preparation, propagation, compounding, assembly, or processing of medical devices...

  3. Utilization of a ts-sacB selection system for the generation of a Mycobacterium avium serovar-8 specific glycopeptidolipid allelic exchange mutant

    OpenAIRE

    Belisle John T; Inamine Julia M; Eckstein Torsten M; Lee Sun-Hwa; Irani Vida R; Maslow Joel N

    2004-01-01

    Abstract Background Mycobacterium avium are ubiquitous environmental organisms and a cause of disseminated infection in patients with end-stage AIDS. The glycopeptidolipids (GPL) of M. avium are proposed to participate in the pathogenesis of this organism, however, establishment of a clear role for GPL in disease production has been limited by the inability to genetically manipulate M. avium. Methods To be able to study the role of the GPL in M. avium pathogenesis, a ts-sacB selection system,...

  4. PRESENCE OF MYCOBACTERIUM AVIUM SUBSP. PARATUBERCULOSIS IN ALPACAS (LAMA PACOS) INHABITING THE CHILEAN ALTIPLANO.

    Science.gov (United States)

    Salgado, Miguel; Sevilla, Iker; Rios, Carolina; Crossley, Jorge; Tejeda, Carlos; Manning, Elizabeth

    2016-03-01

    Mycobacterium avium subsp. paratuberculosis (MAP) is the etiologic agent of paratuberculosis. The organism causes disease in both domestically managed and wild ruminant species. South American camelids have a long, shared history with indigenous people in the Andes. Over the last few decades, increasing numbers of alpacas were exported to numerous countries outside South America. No paratuberculosis surveillance has been reported for these source herds. In this study, individual fecal samples from 85 adult alpacas were collected from six separate herds in the Chilean Altiplano. A ParaTB mycobacterial growth indicator tube (MGIT) liquid culture of each individual fecal sample, followed by real-time polymerase chain reaction (PCR) protocol was used for confirmation. DNA extracts from a subset of confirmed MAP isolates were subjected to mycobacterial interspersed repetitive units-variable number of tandem repeats (MIRU-VNTR) typing. Fifteen alpaca were fecal culture test-positive. Five false-positive culture samples were negative on PCR analysis for Mycobacterium avium subsp. avium (MAA), Mycobacterium bovis (M. bovis), and the 16 S rDNA gene. Three MAP isolates subset-tested belonged to the same MIRU-VNTR type, showing four repeats for TR292 (locus 1) in contrast to the three repeats typical of the MAP reference strain K10. The number of repeats found in the remaining loci was identical to that of the K10 strain. It is not known how nor when MAP was introduced into the alpaca population in the Chilean Altiplano. The most plausible hypothesis to explain the presence of MAP in these indigenous populations is transmission by contact with infected domestic small ruminant species that may on occasion share pastures or range with alpacas. Isolation of this mycobacterial pathogen from such a remote region suggests that MAP has found its way beyond the confines of intensively managed domestic agriculture premises.

  5. Genome-wide sequence variations among Mycobacterium avium subspecies paratuberculosis.

    Directory of Open Access Journals (Sweden)

    Chung-Yi eHsu

    2011-12-01

    Full Text Available Mycobacterium avium subspecies paratuberculosis (M. ap, the causative agent of Johne’s disease (JD, infects many farmed ruminants, wildlife animals and humans. To better understand the molecular pathogenesis of these infections, we analyzed the whole genome sequences of several M. ap and M. avium subspecies avium (M. avium strains isolated from various hosts and environments. Using Next-generation sequencing technology, all 6 M. ap isolates showed a high percentage of homology (98% to the reference genome sequence of M. ap K-10 isolated from cattle. However, 2 M. avium isolates (DT 78 and Env 77 showed significant sequence diversity from the reference strain M. avium 104. The genomes of M. avium isolates DT 78 and Env 77 exhibited only 87% and 40% homology, respectively, to the M. avium 104 reference genome. Within the M. ap isolates, genomic rearrangements (insertions/deletions, Indels were not detected, and only unique single nucleotide polymorphisms (SNPs were observed among the 6 M. ap strains. While most of the SNPs (~100 in M. ap genomes were non-synonymous, a total of ~ 6000 SNPs were detected among M. avium genomes, most of them were synonymous suggesting a differential selective pressure between M. ap and M. avium isolates. In addition, SNPs-based phylo-genomic analysis showed that isolates from goat and Oryx are closely related to the cattle (K-10 strain while the human isolate (M. ap 4B is closely related to the environmental strains, indicating environmental source to human infections. Overall, SNPs were the most common variations among M. ap isolates while SNPs in addition to Indels were prevalent among M. avium isolates. Genomic variations will be useful in designing host-specific markers for the analysis of mycobacterial evolution and for developing novel diagnostics directed against Johne’s disease in animals.

  6. Cerebrospinal fluid neopterin analysis in neuropediatric patients: establishment of a new cut off-value for the identification of inflammatory-immune mediated processes.

    Directory of Open Access Journals (Sweden)

    Marta Molero-Luis

    Full Text Available OBJECTIVE: A high level of cerebrospinal fluid (CSF neopterin is a marker of central nervous system inflammatory-immune mediated processes. We aimed to assess data from 606 neuropediatric patients, describing the clinical and biochemical features of those neurological disorders presenting CSF neopterin values above a new cut-off value that was defined in our laboratory. METHODS: To establish the new CSF neopterin cut-off value, we studied two groups of patients: Group 1 comprised 68 patients with meningoencephalitis, and Group 2 comprised 52 children with a confirmed peripheral infection and no central nervous system involvement. We studied 606 CSF samples from neuropediatric patients who were classified into 3 groups: genetic diagnosis (A, acquired/unknown etiologic neurologic diseases (B and inflammatory-immune mediated processes (C. RESULTS: The CSF neopterin cut-off value was 61 nmol/L. Out of 606 cases, 56 presented a CSF neopterin level above this value. Group C had significantly higher CSF neopterin, protein and leukocyte values than the other groups. Sixteen of twenty-three patients in this group had a CSF neopterin level above the cut-off, whereas three and seven patients presented increased leukocyte and protein values, respectively. A significant association was found among CSF neopterin, proteins and leukocytes in the 606 patients. White matter disturbances were associated with high CSF neopterin concentrations. CONCLUSIONS: Although children with inflammatory-immune mediated processes presented higher CSF neopterin values, patients with other neurological disorders also showed increased CSF neopterin concentrations. These results stress the importance of CSF neopterin analysis for the identification of inflammatory-immune mediated processes.

  7. Establishment study of the in vivo imaging analysis with small animal imaging modalities (micro-PET and micro-SPECT/CT) for bio-drug development

    Energy Technology Data Exchange (ETDEWEB)

    Jang, Beomsu; Park, Sanghyeon; Park, Jeonghoon; Jo, Sungkee; Jung, Uhee; Kim, Seolwha; Lee, Yunjong; Choi, Daeseong

    2011-01-15

    In this study, we established the image acquisition and analysis procedures of micro-PET, SPECT/CT using the experimental animal (mouse) for the development of imaging assessment method for the bio-drug. We examined the micro-SPECT/CT, PET imaging study using the Siemens Inveon micro-multimodality system (SPECT/CT) and micro-PET with {sup 99m}Tc-MDP, DMSA, and {sup 18}F-FDG. SPECT imaging studies using 3 types of pinhole collimators. 5-MWB collimator was used for SPECT image study. To study whole-body distribution, {sup 99m}Tc-MDP SPECT image study was performed. We obtained the fine distribution image. And the CT images was obtained to provide the anatomical information. And then these two types images are fused. To study specific organ uptake, we examined {sup 99}mTc-DMSA SPECT/CT imaging study. We also performed the PET image study using U87MG tumor bearing mice and {sup 18}F-FDG. The overnight fasting, warming and anesthesia with 2% isoflurane pretreatment enhance the tumor image through reducing the background uptake including brown fat, harderian gland and skeletal muscles. Also we got the governmental approval for use of x-ray generator for CT and radioisotopes as sealed and open source. We prepared the draft of process procedure for the experimental animal imaging facility. These research results can be utilized as a basic image study protocols and data for the image assessment of drugs including biological drug.

  8. Chronic mycobacterial meningitis due to Mycobacterium chelonae: a case report.

    Science.gov (United States)

    Salmanzadeh, Shokrallah; Honarvar, Negin; Goodarzi, Hamed; Khosravi, Azar Dokht; Nashibi, Roohangiz; Serajian, Amir Arsalan; Hashemzadeh, Mohammad

    2014-10-01

    We report a case of chronic meningitis due to Mycobacterium chelonae. This organism is a rapidly growing Mycobacterium (RGM) and can be found worldwide in environmental sources such as soil, dust, and water. M. chelonae is an uncommon cause of meningitis; the majority of infections caused by this organism are localized cutaneous or soft tissue infections, and rarely lung infections. The organism is indistinguishable phenotypically, so we applied PCR based on the rpoB gene sequence followed by restriction fragment length polymorphism (RFLP) for molecular identification. The subsequent sequencing of RFLP products revealed 99.7% similarity with M. chelonae.

  9. Function and antigen recognition pattern of L3T4+ T-cell clones from Mycobacterium tuberculosis-immune mice.

    OpenAIRE

    Kaufmann, S H; Flesch, I

    1986-01-01

    T-cell clones were established from Mycobacterium tuberculosis-immunized mice. These clones had the phenotype Thy-1+ L3T4+ Lyt-2- and were restricted by the H-2I-A locus. After antigen stimulation, the T-cell clones secreted interleukin-2 and gamma interferon. Factors produced by these T-cell clones activated normal bone marrow macrophages for antimycobacterial activity in vitro. Furthermore, the T-cell clones could adoptively confer delayed-type hypersensitivity on normal recipient mice. The...

  10. Novel antigens for detection of cell mediated immune responses to Mycobacterium avium subsp. paratuberculosis infection in cattle

    DEFF Research Database (Denmark)

    Mikkelsen, Heidi; Aagaard, Claus; Nielsen, Søren Saxmose;

    2011-01-01

    Paratuberculosis is a chronic infection of the intestine of ruminants caused by Mycobacterium avium subsp. paratuberculosis (MAP). Early stage MAP infection can be detected by measuring specific cell mediated immune responses, using the whole blood interferon-γ (IFN-γ) assay. Available IFN-γ assays...... included blood samples from 26 heifers from a MAP infected herd, collected three times with four to five-week intervals, and blood samples from 60 heifers of a non-infected herd collected once. Heifers of the non-infected herd were used to establish cut-off values for each antigen. The case definition...

  11. Novel Real-Time Simultaneous Amplification and Testing Method To Accurately and Rapidly Detect Mycobacterium tuberculosis Complex

    OpenAIRE

    Cui, Zhenling; Wang, Yongzhong; Fang, Liang; Zheng, Ruijuan; Huang, Xiaochen; Liu, Xiaoqin; Zhang, Gang; Rui, Dongmei; Ju, Jinliang; Hu, Zhongyi

    2012-01-01

    The aim of this study was to establish and evaluate a simultaneous amplification and testing method for detection of the Mycobacterium tuberculosis complex (SAT-TB assay) in clinical specimens by using isothermal RNA amplification and real-time fluorescence detection. In the SAT-TB assay, a 170-bp M. tuberculosis 16S rRNA fragment is reverse transcribed to DNA by use of Moloney murine leukemia virus (M-MLV) reverse transcriptase, using specific primers incorporating the T7 promoter sequence, ...

  12. 浅析绵阳生态城市建设%Primary Analysis for the Establishment of Ecological City of Mianyang

    Institute of Scientific and Technical Information of China (English)

    张新合

    2005-01-01

    This paper probes into the establishment of ecological city of Mianyang and directive thoughts, measures,and countermoves are put forward based on analyzing the current situation about ecological environment of Mianyang City.

  13. Analysis on the Influencing Factors of the Quality Traceable System Established by Edible Agricultural Products Enterprises—Taking Sichuan as an example

    OpenAIRE

    Xie, Xiao; Wu, Xiu-min; Zhao, Zhi-jing

    2011-01-01

    According to the investigation data from 81 edible agricultural products enterprises in Sichuan Province, the influence factors of establishing quality tracing system are empirically analyzed from four aspects, including the enterprise features, the attitudes of operators, management and market competition by using the Logistic model. The results show that the enterprise can establish the quality tracing system after the comprehensive function of a series of internal and external factors. The...

  14. Drug resistant Mycobacterium tuberculosis of the Beijing genotype does not spread in Sweden.

    Directory of Open Access Journals (Sweden)

    Solomon Ghebremichael

    Full Text Available BACKGROUND: Drug resistant (DR and multi-drug resistant (MDR tuberculosis (TB is increasing worldwide. In some parts of the world 10% or more of new TB cases are MDR. The Beijing genotype is a distinct genetic lineage of Mycobacterium tuberculosis, which is distributed worldwide, and has caused large outbreaks of MDR-TB. It has been proposed that certain lineages of M. tuberculosis, such as the Beijing lineage, may have specific adaptive advantages. We have investigated the presence and transmission of DR Beijing strains in the Swedish population. METHODOLOGY/PRINCIPAL FINDINGS: All DR M. tuberculosis complex isolates between 1994 and 2008 were studied. Isolates that were of Beijing genotype were investigated for specific resistance mutations and phylogenetic markers. Seventy (13% of 536 DR strains were of Beijing genotype. The majority of the patients with Beijing strains were foreign born, and their country of origin reflects the countries where the Beijing genotype is most prevalent. Multidrug-resistance was significantly more common in Beijing strains than in non-Beijing strains. There was a correlation between the Beijing genotype and specific resistance mutations in the katG gene, the mabA-inhA-promotor and the rpoB gene. By a combined use of RD deletions, spoligotyping, IS1547, mutT gene polymorphism and Rv3135 gene analysis the Beijing strains could be divided into 11 genomic sublineages. Of the patients with Beijing strains 28 (41% were found in altogether 10 clusters (2-5 per cluster, as defined by RFLP IS6110, while 52% of the patients with non-Beijing strains were in clusters. By 24 loci MIRU-VNTR 31 (45% of the patients with Beijing strains were found in altogether 7 clusters (2-11 per cluster. Contact tracing established possible epidemiological linkage between only two patients with Beijing strains. CONCLUSIONS/SIGNIFICANCE: Although extensive outbreaks with non-Beijing TB strains have occurred in Sweden, Beijing strains have not

  15. Analysis on multi-drug resistance of Mycobacterium tuberculosis in Xinjiang%新疆地区93株耐多药结核分枝杆菌耐药情况分析

    Institute of Scientific and Technical Information of China (English)

    李君莲; 张敬蕊; 李桂莲; 李东军; 张媛媛; 万康林; 綦迎成

    2012-01-01

    Objective To analyze multi-drug resistance (MDR) of Mycobacterium tuberculosis (M. tuberculosis) isolated from Xinjiang, to provide evidence for the treatment of multi-drug resistant tuberculosis (MDR-TB). Methods A total of 93 MDR M. tuberculosis clinical strains which were identified by routine drug susceptibility testing with proportion method were collected from 2009 to 2011 for the drug sensitivity to 14 drugs including streptomycin, ethambutol, ofloxacin, levofloxacin, amikacin, capreomycin, protionamide, pasiniazide, clarithromycin, ciprofloxacin, rifabutin, clofazimine, linezolid and sulfamethoxazole using HX-21 bacterial identification and drug susceptibility test plate. Results For the first-line drugs, the resistant rates to streptomycin and ethambutol were 48. 39% (45/93) and 15.05% (14/93), and for the second-line drugs, the highest resistance rate was 51.61% (48/93) to rifabutin, while the lowest rate was 1.08% (1/93) to capreomycin. There were 10 (10. 75%) strains resistant to both isoniazid and rifampicin. Except for isoniazid and rifampicin, the highest double-resistant rate to other drugs was 25.81% (24/93). The cross-resistant rates to aminoglycosides streptomycin and amikacin, to ofloxacin and ciprofloxacin and to rifampicin and rifabutm were 6.45% (6/93), 12.90% (12/93) and 51.61% (48/93) respectively. There were 6 strains identified as extensive-drug resistant which accounted for 6. 45% (6/93). Conclusions There are relatively high multi-drug and cross-drug resistant rates among clinically isolated M. tuberculosis strains in Xinjiang especially to. those first and second-line drugs and to rifampicin and rifabutin. Therefore patients with MDR-TB should be treated carefully to avoid drug resistance and cross-resistance.%目的 分析新疆地区耐多药结核分枝杆菌的耐药情况,为指导耐多药结核病的治疗提供依据.方法 收集新疆地区2009-2011年经常规药敏试验发现的耐多药结核分枝杆菌(MDR-TB

  16. Analysis of the drug-resistant situation and antibiogram of Mycobacterium tuberculosis in Fanyu%番禺地区结核分枝杆菌耐药状况及耐药谱分析

    Institute of Scientific and Technical Information of China (English)

    郭廷学; 陈敏

    2015-01-01

    目的:探析番禺地区结核分枝杆菌的耐药情况与耐药谱。方法选择2011年2月至2014年2月期间番禺地区就诊的结核病患者1033例,并对其分离株进行菌种鉴定。检测其对利福平( RIF)、异烟肼( INH)、链霉素( SM)及乙胺丁醇( EMB)的耐药性。并进一步分析对INH耐药的结核分枝杆菌的耐药谱分布情况。结果 1033株结核菌株中,总耐药率为42.01%(434/1033),其中,初始患者耐药率低于复治患者(40.32%比48.20%),两者耐药率具有统计学差异(χ2=4.440,P =0.035)。男性患者耐药率低于女性患者(37.18%比57.55%),两者耐药率具有统计学差异(χ2=31.826,P <0.01)。30˜60岁年龄段患者耐药率最高,<30岁、30˜60岁和>60岁3个年龄段耐药率(28.78%比48.31%比27.68%)比较差异有统计学意义(χ2=48.852,P <0.01)。所检测的RIF、INH、SM及EMB耐药顺序为INH (14.98%)>RIF(13.36%)>SM(9.11%)>EMB(4.56%)。155株INH耐药株中,同时耐RIF的有46(29.68%)株,耐多药率为4.45%(46/1033)。结论番禺地区结核分枝杆菌的耐药情况较严重,与患者年龄、性别等有关,应加强对耐药杆菌的检测,采取有效的防控措施。%Objective To investigate drug -resistant situation and antibiogram of Mycobacterium tuberculosis( M. tuberculosis)in Fanyu. Methods During the time of February 2011 to February 2014,1 033 tuberculosis patients were selected and the isolates obtained from the patients were identified. The resistance of these isolates to rifampicin,isoniazid,streptomycin and ethambutol were tested and the antibio-gram were analyzed. Results The total drug-resistance rate of M. tuberculosis isolates was 42. 01%(434/1 033). The initial drug-resist-ance rate(40. 32%)was lower than acquired drug-resistance rate(48. 20%),the difference was statistically significant(χ2 =4. 440,P =0. 035). The drug-resistance rate

  17. Clinical analysis of 114 AIDS patients with positive blood culture for Mycobacterium tuberculosis%114例血培养结核分枝杆菌阳性AIDS患者临床分析

    Institute of Scientific and Technical Information of China (English)

    李雪琴; 刘升; 邓建宁; 黄爱春; 卢祥婵

    2013-01-01

    目的 分析血培养结核分枝杆菌阳性AIDS患者的临床资料,为AIDS合并结核病的诊治提供依据.方法 回顾性分析114例血培养结核分枝杆菌阳性AIDS患者的临床资料.结果 114例中男95例,女19例,从出现临床症状到就诊的中位时间为36d.临床表现主要有发热(102例)、消瘦(97例)、咳嗽(78例)、咳痰(63例)、盗汗(51例)、腹泻(45例)和浅表淋巴结肿大(45例).CD4+ T淋巴细胞为4~501个/mm3,其中<100个/mm3有105例(92.1%).结核病耐药41例(36.0%),其中单耐药11例(9.6%),耐多药及广泛耐药14例(12.3%).腹主动脉旁淋巴结肿大44例(38.6%).胸腔积液31例(27.2%),心包积液18例(15.8%).胸片检查异常99例(86.8%).经治疗后,35例(30.7%)死亡,其中33例(94.3%)CD4+T淋巴细胞<100个/mm3.结论 血培养结核分枝杆菌阳性多见于CD4+ T淋巴细胞<100个/mm3的男性AIDS患者,病程约1个月,临床表现主要为发热、消瘦、咳嗽、咳痰及淋巴结肿大,可有胸腔积液,大部分患者肺部有病灶.结核病耐药发生率高,晚期患者病死率高.%Objective To analyze the clinical data of AIDS patients with positive blood culture for Mycobacterium tuberculosis,so as to provide an evidence for the diagnosis and treatment of AIDS complicated by tuberculosis.Methods The chnical data of 114 AIDS patients with positive blood culture for M.tuberculosis were analyzed retrospectively.Results Of 114 AIDS patients,95 were males,and 19 were females.The median duration from the presence of clinical symptoms to clinic visit was 36 days.The main clinical manifestations were fever in 102 patients,marasmus in 97 patients,cough in 78 patients,expectoration in 63 patients,night sweat in 51 patients,diarrhea in 45 patients and superficial lymph node enlargement in 45 patients.The CD4+ T lymphocyte count ranged from 4 cells/mm3 to 501 cells/mm3,and 105 patients (92.1%) had a CD4+ T lymphocyte count of less than 100 cells/mm3

  18. Inhibiting Mycobacterium tuberculosis within and without.

    Science.gov (United States)

    Cole, Stewart T

    2016-11-01

    Tuberculosis remains a scourge of global health with shrinking treatment options due to the spread of drug-resistant strains of Mycobacterium tuberculosis Intensive efforts have been made in the past 15 years to find leads for drug development so that better, more potent drugs inhibiting new targets could be produced and thus shorten treatment duration. Initial attempts focused on repurposing drugs that had been developed for other therapeutic areas but these agents did not meet their goals in clinical trials. Attempts to find new lead compounds employing target-based screens were unsuccessful as the leads were inactive against M. tuberculosis Greater success was achieved using phenotypic screening against live tubercle bacilli and this gave rise to the drugs bedaquiline, pretomanid and delamanid, currently in phase III trials. Subsequent phenotypic screens also uncovered new leads and targets but several of these targets proved to be promiscuous and inhibited by a variety of seemingly unrelated pharmacophores. This setback sparked an interest in alternative screening approaches that mimic the disease state more accurately. Foremost among these were cell-based screens, often involving macrophages, as these should reflect the bacterium's niche in the host more faithfully. A major advantage of this approach is its ability to uncover functions that are central to infection but not necessarily required for growth in vitro For instance, inhibition of virulence functions mediated by the ESX-1 secretion system severely attenuates intracellular M. tuberculosis, preventing intercellular spread and ultimately limiting tissue damage. Cell-based screens have highlighted the druggability of energy production via the electron transport chain and cholesterol metabolism. Here, I review the scientific progress and the pipeline, but warn against over-optimism due to the lack of industrial commitment for tuberculosis drug development and other socio-economic factors.This article is

  19. Mycolic Acid Cyclopropanation is Essential for Viability, Drug Resistance, and Cell Wall Integrity of Mycobacterium tuberculosis

    Energy Technology Data Exchange (ETDEWEB)

    Barkan, Daniel; Liu, Zhen; Sacchettini, James C.; Glickman, Michael S.; (MSKCC); (TAM)

    2009-12-01

    Mycobacterium tuberculosis infection remains a major global health problem complicated by escalating rates of antibiotic resistance. Despite the established role of mycolic acid cyclopropane modification in pathogenesis, the feasibility of targeting this enzyme family for antibiotic development is unknown. We show through genetics and chemical biology that mycolic acid methyltransferases are essential for M. tuberculosis viability, cell wall structure, and intrinsic resistance to antibiotics. The tool compound dioctylamine, which we show acts as a substrate mimic, directly inhibits the function of multiple mycolic acid methyltransferases, resulting in loss of cyclopropanation, cell death, loss of acid fastness, and synergistic killing with isoniazid and ciprofloxacin. These results demonstrate that mycolic acid methyltransferases are a promising antibiotic target and that a family of virulence factors can be chemically inhibited with effects not anticipated from studies of each individual enzyme.

  20. Broncho-pleural fistula with hydropneumothorax at CT: Diagnostic implications in mycobacterium avium complex lung disease with pleural involvement

    Energy Technology Data Exchange (ETDEWEB)

    Yoon, Hyun Jung; Chung, Myung Jin; Lee, Kyung Soo; Park, Hye Yun; Koh, Won Jung [Samsung Medical Center, Sungkyunkwan University School of Medicine, Seoul (Korea, Republic of); Kim, Jung Soo [Division of Pulmonary and Critical Care Medicine, Department of Internal Medicine, Inha University Hospital, Inha University School of Medicine, Incheon (Korea, Republic of)

    2016-04-15

    To determine the patho-mechanism of pleural effusion or hydropneumothorax in Mycobacterium avium complex (MAC) lung disease through the computed tomographic (CT) findings. We retrospectively collected data from 5 patients who had pleural fluid samples that were culture-positive for MAC between January 2001 and December 2013. The clinical findings were investigated and the radiological findings on chest CT were reviewed by 2 radiologists. The 5 patients were all male with a median age of 77 and all had underlying comorbid conditions. Pleural fluid analysis revealed a wide range of white blood cell counts (410-100690/µL). The causative microorganisms were determined as Mycobacterium avium and Mycobacterium intracellulare in 1 and 4 patients, respectively. Radiologically, the peripheral portion of the involved lung demonstrated fibro-bullous changes or cavitary lesions causing lung destruction, reflecting the chronic, insidious nature of MAC lung disease. All patients had broncho-pleural fistulas (BPFs) and pneumothorax was accompanied with pleural effusion. In patients with underlying MAC lung disease who present with pleural effusion, the presence of BPFs and pleural air on CT imaging are indicative that spread of MAC infection is the cause of the effusion.

  1. The virtual observatory service TheoSSA: Establishing a database of synthetic stellar flux standards . II. NLTE spectral analysis of the OB-type subdwarf Feige 110

    Science.gov (United States)

    Rauch, T.; Rudkowski, A.; Kampka, D.; Werner, K.; Kruk, J. W.; Moehler, S.

    2014-06-01

    Context. In the framework of the Virtual Observatory (VO), the German Astrophysical VO (GAVO) developed the registered service TheoSSA (Theoretical Stellar Spectra Access). It provides easy access to stellar spectral energy distributions (SEDs) and is intended to ingest SEDs calculated by any model-atmosphere code, generally for all effective temperatures, surface gravities, and elemental compositions. We will establish a database of SEDs of flux standards that are easily accessible via TheoSSA's web interface. Aims: The OB-type subdwarf Feige 110 is a standard star for flux calibration. State-of-the-art non-local thermodynamic equilibrium stellar-atmosphere models that consider opacities of species up to trans-iron elements will be used to provide a reliable synthetic spectrum to compare with observations. Methods: In case of Feige 110, we demonstrate that the model reproduces not only its overall continuum shape from the far-ultraviolet (FUV) to the optical wavelength range but also the numerous metal lines exhibited in its FUV spectrum. Results: We present a state-of-the-art spectral analysis of Feige 110. We determined , log g = 6.00 ± 0.20, and the abundances of He, N, P, S, Ti, V, Cr, Mn, Fe, Co, Ni, Zn, and Ge. Ti, V, Mn, Co, Zn, and Ge were identified for the first time in this star. Upper abundance limits were derived for C, O, Si, Ca, and Sc. Conclusions: The TheoSSA database of theoretical SEDs of stellar flux standards guarantees that the flux calibration of astronomical data and cross-calibration between different instruments can be based on models and SEDs calculated with state-of-the-art model-atmosphere codes. Based on observations with the NASA/ESA Hubble Space Telescope, obtained at the Space Telescope Science Institute, which is operated by the Association of Universities for Research in Astronomy, Inc., under NASA contract NAS5-26666. Based on observations made with the NASA-CNES-CSA Far Ultraviolet Spectroscopic Explorer. Table 2, Figs. 3 and

  2. Characterization of Mycobacterium tuberculosis Beijing isolates from the Mediterranean area

    Directory of Open Access Journals (Sweden)

    Samper S

    2010-05-01

    Full Text Available Abstract Background The Beijing lineage of Mycobacterium tuberculosis is causing concern due to its global distribution and its involvement in severe outbreaks. Studies focused on this lineage are mainly restricted to geographical settings where its prevalence is high, whereas those in other areas are scarce. In this study, we analyze Beijing isolates in the Mediterranean area, where this lineage is not prevalent and is mainly associated with immigrant cases. Results Only 1% (N = 26 of the isolates from two population-based studies in Spain corresponded to Beijing strains, most of which were pan-susceptible and from Peruvian and Ecuadorian patients. Restriction fragment length polymorphism typing with the insertion sequence IS6110 identified three small clusters (2-3 cases. Mycobacterial interspersed repetitive unit-variable number tandem repeat typing (MIRU-15 offered low discriminatory power, requiring the introduction of five additional loci. A selection of the Beijing isolates identified in the Spanish sample, together with a sample of Beijing strains from Italy, to broaden the analysis context in the Mediterranean area, were assayed in an infection model with THP-1 cells. A wide range of intracellular growth rates was observed with only two isolates showing an increased intracellular replication, in both cases associated with contained production of TNF-α. No correlation was observed between virulence and the Beijing phylogenetic group, clustered/orphan status, or resistance. The Beijing strain responsible for extensive spread on Gran Canaria Island was also identified in Madrid, but did not lead to secondary cases and did not show high infectivity in the infection model. Conclusions The Beijing lineage in our area is a non-homogeneous family, with only certain highly virulent representatives. The specific characterization of Beijing isolates in different settings could help us to accurately identify the virulent representatives before

  3. Characterization of IS1245 for Strain Typing of Mycobacterium avium

    Science.gov (United States)

    Pestel-Caron, Martine; Arbeit, Robert D.

    1998-01-01

    IS1245 is an insertion element widely prevalent among isolates of Mycobacterium avium. We used PvuII Southern blots to analyze IS1245 polymorphisms among 159 M. avium isolates (141 clinical isolates from 40 human immunodeficiency virus-infected patients plus 18 epidemiologically related environmental isolates) that represented 40 distinct M. avium strains, as resolved by previous studies by pulsed-field gel electrophoresis (PFGE). All 40 strains carried DNA homologous to IS1245 and thus were typeable. Twenty-five (63%) strains had ≥10 copies of the element, 6 (15%) had 4 to 9 copies, and 9 (23%) had only 1 to 3 copies. Among the last group of nine strains (each of which was distinct by PFGE analysis), IS1245 typing resolved only four patterns and thus provided poor discriminatory power. To evaluate the in vivo stability of IS1245, we analyzed 32 strains for which sets of 2 to 19 epidemiologically related isolates were available. For 19 (59%) of these sets, all isolates representing the same strain had indistinguishable IS1245 patterns. Within eight (25%) sets, one or more isolates had IS1245 patterns that differed by one or two fragments from the modal pattern for the isolates of that strain. Five (16%) sets included isolates whose patterns differed by three or more fragments; on the basis of IS1245 typing those isolates would have been designated distinct strains. IS1245 was stable during in vitro passage, suggesting that the variations observed represented natural translocations of the element. IS1245 provides a useful tool for molecular strain typing of M. avium but may have limitations for analyzing strains with low copy numbers or for resolving extended epidemiologic relationships. PMID:9650925

  4. Biochemical characterization of recombinant phosphoglucose isomerase of Mycobacterium tuberculosis

    International Nuclear Information System (INIS)

    Phosphoglucose isomerase (PGI) is a well-characterized ubiquitous enzyme involved in the glycolytic pathway. It catalyzes the reversible isomerization of D-glucopyranose-6-phosphate and D-fructofuranose-6-phosphate and is present in all living cells. However, there is interspecies variation at the level of the primary structure which sometimes produces heterogeneity at the structural and functional levels. In order to evaluate and characterize the mycobacterial PGI, the gene encoding the PGI from Mycobacterium tuberculosis H37Rv was cloned in pET-22b(+) vector and expressed in Escherichia coli. The target DNA was PCR amplified from the bacterial artificial chromosome using specific primers and cloned under the control of T7 promoter. Upon induction with IPTG, the recombinant PGI (rPGI) expressed partly as soluble protein and partly as inclusion bodies. The rPGI from the soluble fraction was purified to near homogeneity by ion-exchange chromatography. Mass spectrum analysis of the purified rPGI revealed its mass to be 61.45 kDa. The purified rPGI was enzymatically active and the specific activity was 600 U/mg protein. The K m of rPGI was determined to be 0.318 mM for fructose-6-phosphate and the K i was 0.8 mM for 6-phosphogluconate. The rPGI exhibited optimal activity at 37 deg C and pH 9.0, and did not require mono- or divalent cations for its activity

  5. Molecular epidemiology of Mycobacterium tuberculosis in Gansu province of China

    Institute of Scientific and Technical Information of China (English)

    TIAN Li-li; ZHU Bing-dong; SI Hong-yan; MU Tao-jun; FAN Wen-bing; WANG Jing; JIANG Wei-min; LI Qing; YANG Biao; ZHANG Ying

    2012-01-01

    Background Mycobacterial interspersed repetitive units-variable number tandem repeat (MIRU-VNTR) and Beijing family typing based on detecting the deletion of RD105 sequence are two common genotyping methods used to study the molecular epidemiologic characteristics of Mycobacterium (M.) tuberculosis.We collected 218 strains of M.tuberculosis between 2004 and 2006 in the Linxia Hui Autonomous Prefecture of Gansu province in Northwest China.Methods MIRU-VNTR analysis and Beijing family typing based on detecting the deletion of RD105 sequence were used to type the 218 strains,and their typing power was evaluated to look for practical and efficient genotyping methods suitable for the region.Results The MIRU typing yielded 115 distinct genotypes,including 98 unique isolates and 17 different clusters containing 120 isolates (55.05%); the cluster rate was 47.25%.By detecting the deletion of RD105 sequence,188 of 218 (86.23%) isolates belonged to Beijing family.Combination of Beijing family typing and MIRU typing yielded 118 distinct patterns,including 101 unique isolates and 17 clusters containing 117 isolates (54.13%).The largest cluster contained 58 strains with MIRU genotype of 223325173533 which contained 50 strains belonging to Beijing family and 8 strains belonging to non-Beijing family.Conclusions The Beijing family strains occupied a large proportion and the Beijing family MIRU genotype 223325173533 is a dominant strain in Linxia of Gansu.Combining detecting the deletion of RD105 and MIRU typing together provides a simple,fast,and effective method which is low in cost and might be practical and suitable for M.tuberculosis genotyping in China.

  6. Optical mapping of the Mycobacterium avium subspecies paratuberculosis genome

    Directory of Open Access Journals (Sweden)

    Schwartz David C

    2009-01-01

    Full Text Available Abstract Background Infection of cattle with Mycobacterium avium subspecies paratuberculosis (M. ap causes severe economic losses to the dairy industry in the USA and worldwide. In an effort to better examine diversity among M. ap strains, we used optical mapping to profile genomic variations between strains of M. ap K-10 (sequenced strain and M. ap ATCC 19698 (type strain. Results The assembled physical restriction map of M. ap ATCC 19698 showed a genome size of 4,839 kb compared to the sequenced K-10 genome of 4,830 kb. Interestingly, alignment of the optical map of the M. ap ATCC 19698 genome to the complete M. ap K-10 genome sequence revealed a 648-kb inversion around the origin of replication. However, Southern blotting, PCR amplification and sequencing analyses of the inverted region revealed that the genome of M. ap K-10 differs from the published sequence in the region starting from 4,197,080 bp to 11,150 bp, spanning the origin of replication. Additionally, two new copies of the coding sequences > 99.8% were identified, identical to the MAP0849c and MAP0850c genes located immediately downstream of the MAP3758c gene. Conclusion The optical map of M. ap ATCC 19698 clearly indicated the miss-assembly of the sequenced genome of M. ap K-10. Moreover, it identified 2 new genes in M. ap K-10 genome. This analysis strongly advocates for the utility of physical mapping protocols to complement genome sequencing projects.

  7. Molecular Epidemiology of Mycobacterium bovis in Humans and Cattle.

    Science.gov (United States)

    El-Sayed, A; El-Shannat, S; Kamel, M; Castañeda-Vazquez, M A; Castañeda-Vazquez, H

    2016-06-01

    Bovine tuberculosis (bTB), caused by Mycobacterium bovis (M. bovis), is a serious re-emerging disease in both animals and humans. The evolution of the Multi- and Extensively drug-resistant M. bovis strains (MDR-TB and XDR-TB) represents a global threat to public health. Worldwide, the disease is responsible for great economic losses in the veterinary field, serious threat to the ecosystem, and about 3.1% of human TB cases, up to 16% in Tanzania. Only thorough investigation to understand the pathogen's epidemiology can help in controlling the disease and minimizing its threat. For this purpose, various tools have been developed for use in advanced molecular epidemiological studies of bTB, either alone or in combination with standard conventional epidemiological approaches. These techniques enable the analysis of the intra- and inter-species transmission dynamics of bTB. The delivered data can reveal detailed insights into the source of infection, correlations among human and bovine isolates, strain diversity and evolution, spread, geographical localization, host preference, tracing of certain virulence factors such as antibiotic resistance genes, and finally the risk factors for the maintenance and spread of M. bovis. They also allow for the determination of epidemic and endemic strains. This, in turn, has a significant diagnostic impact and helps in vaccine development for bTB eradication programs. The present review discusses many topics including the aetiology, epidemiology and importance of M. bovis, the prevalence of bTB in humans and animals in various countries, the molecular epidemiology of M. bovis, and finally applied molecular epidemiological techniques. PMID:26684712

  8. Modeling tuberculous meningitis in zebrafish using Mycobacterium marinum

    Directory of Open Access Journals (Sweden)

    Lisanne M. van Leeuwen

    2014-09-01

    Full Text Available Tuberculous meningitis (TBM is one of the most severe extrapulmonary manifestations of tuberculosis, with a high morbidity and mortality. Characteristic pathological features of TBM are Rich foci, i.e. brain- and spinal-cord-specific granulomas formed after hematogenous spread of pulmonary tuberculosis. Little is known about the early pathogenesis of TBM and the role of Rich foci. We have adapted the zebrafish model of Mycobacterium marinum infection (zebrafish–M. marinum model to study TBM. First, we analyzed whether TBM occurs in adult zebrafish and showed that intraperitoneal infection resulted in granuloma formation in the meninges in 20% of the cases, with occasional brain parenchyma involvement. In zebrafish embryos, bacterial infiltration and clustering of infected phagocytes was observed after infection at three different inoculation sites: parenchyma, hindbrain ventricle and caudal vein. Infection via the bloodstream resulted in the formation of early granulomas in brain tissue in 70% of the cases. In these zebrafish embryos, infiltrates were located in the proximity of blood vessels. Interestingly, no differences were observed when embryos were infected before or after early formation of the blood-brain barrier (BBB, indicating that bacteria are able to cross this barrier with relatively high efficiency. In agreement with this observation, infected zebrafish larvae also showed infiltration of the brain tissue. Upon infection of embryos with an M. marinum ESX-1 mutant, only small clusters and scattered isolated phagocytes with high bacterial loads were present in the brain tissue. In conclusion, our adapted zebrafish–M. marinum infection model for studying granuloma formation in the brain will allow for the detailed analysis of both bacterial and host factors involved in TBM. It will help solve longstanding questions on the role of Rich foci and potentially contribute to the development of better diagnostic tools and therapeutics.

  9. Mycobacterium tuberculosis whole genome sequencing and protein structure modelling provides insights into anti-tuberculosis drug resistance

    KAUST Repository

    Phelan, Jody

    2016-03-23

    Background Combating the spread of drug resistant tuberculosis is a global health priority. Whole genome association studies are being applied to identify genetic determinants of resistance to anti-tuberculosis drugs. Protein structure and interaction modelling are used to understand the functional effects of putative mutations and provide insight into the molecular mechanisms leading to resistance. Methods To investigate the potential utility of these approaches, we analysed the genomes of 144 Mycobacterium tuberculosis clinical isolates from The Special Programme for Research and Training in Tropical Diseases (TDR) collection sourced from 20 countries in four continents. A genome-wide approach was applied to 127 isolates to identify polymorphisms associated with minimum inhibitory concentrations for first-line anti-tuberculosis drugs. In addition, the effect of identified candidate mutations on protein stability and interactions was assessed quantitatively with well-established computational methods. Results The analysis revealed that mutations in the genes rpoB (rifampicin), katG (isoniazid), inhA-promoter (isoniazid), rpsL (streptomycin) and embB (ethambutol) were responsible for the majority of resistance observed. A subset of the mutations identified in rpoB and katG were predicted to affect protein stability. Further, a strong direct correlation was observed between the minimum inhibitory concentration values and the distance of the mutated residues in the three-dimensional structures of rpoB and katG to their respective drugs binding sites. Conclusions Using the TDR resource, we demonstrate the usefulness of whole genome association and convergent evolution approaches to detect known and potentially novel mutations associated with drug resistance. Further, protein structural modelling could provide a means of predicting the impact of polymorphisms on drug efficacy in the absence of phenotypic data. These approaches could ultimately lead to novel resistance

  10. Blood Establishment Registration Database

    Data.gov (United States)

    U.S. Department of Health & Human Services — This application provides information for active, inactive, and pre-registered firms. Query options are by FEI, Applicant Name, Establishment Name, Other Names,...

  11. Establishment of WTGs

    International Nuclear Information System (INIS)

    This paper briefly outlines the reasons for establishing a wind turbine generator and the consequent disadvantages. Consideration is then given to the determining factors affecting the economics of wind turbine generators with special reference to Denmark. (UK)

  12. Mycobacterium tuberculosis population structures differ significantly on two Indonesian Islands.

    NARCIS (Netherlands)

    Parwati, I.; Crevel, R. van; Sudiro, M.; Alisjahbana, B.; Pakasi, T.; Kremer, K.; Zanden, A. van der; Soolingen, D. van

    2008-01-01

    Comparison of Mycobacterium tuberculosis genotype distributions in different areas might help to find determinants of the emergence of certain genotypes, such as the Beijing family. In this study, M. tuberculosis isolates originating from patients from two Indonesian islands were genotyped, and poss

  13. Sensitivity of Mycobacterium bovis to common beef processing interventions

    Science.gov (United States)

    Introduction. Cattle infected with Mycobacterium bovis, the causative agent of bovine tuberculosis and a relevant zoonosis to humans, may be sent to slaughter before diagnosis of infection because of slow multiplication of the pathogen. Purpose. This study evaluates multiple processing interventi...

  14. Resistance of Mycobacterium chelonei-like organisms to formaldehyde.

    OpenAIRE

    Hayes, P S; McGiboney, D L; Band, J. D.; Feeley, J C

    1982-01-01

    Mycobacterium chelonei-like organisms have been isolated from patients in two outbreaks of peritonitis involving chronic peritoneal dialysis machines routinely disinfected with 2 to 3% formaldehyde. Susceptibility studies revealed that water-adapted M. chelonei-like organism strains could survive 2 h of exposure to 10% formaldehyde.

  15. Osteomyelitis Because of Mycobacterium Xenopi in an Immunocompetent Child.

    Science.gov (United States)

    Kuntz, Martin; Seidl, Maximilian; Henneke, Philipp

    2016-01-01

    We present the case of a 6-year-old, immunocompetent boy with chronic osteomyelitis of the calcaneus caused by Mycobacterium xenopi. Of note, typical histopathology was not visible on the first biopsy and developed only later over a period of 6 weeks, highlighting the difficult differential diagnosis of osteomyelitis caused by nontuberculous mycobacteria. PMID:26418244

  16. Mycobacterium bovis hip bursitis in a lung transplant recipient.

    Science.gov (United States)

    Dan, J M; Crespo, M; Silveira, F P; Kaplan, R; Aslam, S

    2016-02-01

    We present a report of extrapulmonary Mycobacterium bovis infection in a lung transplant recipient. M. bovis is acquired predominantly by zoonotic transmission, particularly from consumption of unpasteurized foods. We discuss epidemiologic exposure, especially as relates to the Mexico-US border, clinical characteristics, resistance profile, and treatment. PMID:26671334

  17. Mycobacterium marinum Infection After Exposure to Coal Mine Water.

    Science.gov (United States)

    Huaman, Moises A; Ribes, Julie A; Lohr, Kristine M; Evans, Martin E

    2016-01-01

    Mycobacterium marinum infection has been historically associated with exposure to aquariums, swimming pools, fish, or other marine fauna. We present a case of M marinum left wrist tenosynovitis and elbow bursitis associated with a puncture injury and exposure to coal mine water in Illinois. PMID:26835478

  18. Transmissie van Mycobacterium bovis tussen mens en dier

    NARCIS (Netherlands)

    Vries, de G.; Beer, de J.; Bakker, D.; Soolingen, D.

    2015-01-01

    Nederland is officieel vrij van rundertuberculose. Toch komt af en toe nog Mycobacterium bovis-tuberculose voor bij relatief jonge autochtone Nederlanders. Ook zijn er recent nog wel boviene-uitbraken geweest. Dat roept de vraag op of er ook nu nog transmissie is van M.bovis tussen mens en dier. Daa

  19. Serodiagnosis of Mycobacterium abscessus complex infection in cystic fibrosis

    DEFF Research Database (Denmark)

    Qvist, Tavs; Pressler, Tania; Taylor-Robinson, David;

    2015-01-01

    Early signs of pulmonary disease with Mycobacterium abscessus complex (MABSC) can be missed in patients with cystic fibrosis (CF). A serological method could help stratify patients according to risk. The objective of this study was to test the diagnostic accuracy of a novel method for investigating...

  20. Early clearance of Mycobacterium tuberculosis: a new frontier in prevention

    NARCIS (Netherlands)

    Verrall, A.J.; Netea, M.G.; Alisjahbana, B.; Hill, P.C.; Crevel, R. van

    2014-01-01

    Early clearance (EC) is the successful eradication of inhaled Mycobacterium tuberculosis before an adaptive immune response develops. Evidence for EC comes from case contact studies that consistently show that a proportion of heavily exposed individuals do not develop M. tuberculosis infection. Furt

  1. In Vitro Killing of Mycobacterium ulcerans by Acidified Nitrite

    Science.gov (United States)

    Phillips, R.; Kuijper, S.; Benjamin, N.; Wansbrough-Jones, M.; Wilks, M.; Kolk, A. H. J.

    2004-01-01

    Mycobacterium ulcerans, which causes Buruli ulcer, was exposed to acidified nitrite or to acid alone for 10 or 20 min. Killing was rapid, and viable counts were reduced below detectable limits within 10 min of exposure to 40 mM acidified nitrite. M. ulcerans is highly susceptible to acidified nitrite in vitro. PMID:15273132

  2. Mycobacterium avium infection improved by microbial substitution of fungal infection

    OpenAIRE

    Yano, Shuichi

    2010-01-01

    We reported a case of Mycobacterium avium infection in which disease activity appeared to have been suppressed after fungal infection. After the increase in β-D-glucan, her symptoms of fever and chest pain disappeared. We think this phenomenon may be microbial substitution and mild fungal infection may improve the activity due to M avium.

  3. Differentiation of Mycobacterium chelonei from M. fortuitum by ciprofloxacin susceptibility.

    OpenAIRE

    Collins, C. H.; Yates, M D; Uttley, A H

    1985-01-01

    Seventy-five strains of Mycobacterium fortuitum were inhibited by 3.0 mg/l ciprofloxacin but 36 strains of M. chelonei were resistant. The results correlated well with those obtained by the nitratase test. The ciprofloxacin sensitivity test is a useful supplement to the tests used to identify these two species.

  4. Complete Genome Sequence of Mycobacterium ulcerans subsp. shinshuense

    Science.gov (United States)

    Yoshida, Mitsunori; Nakanaga, Kazue; Ogura, Yoshitoshi; Toyoda, Atsushi; Ooka, Tadasuke; Kazumi, Yuko; Mitarai, Satoshi; Ishii, Norihisa; Hayashi, Tetsuya

    2016-01-01

    Mycobacterium ulcerans subsp. shinshuense produces mycolactone and causes Buruli ulcer. Here, we report the complete sequence of its genome, which comprises a 5.9-Mb chromosome and a 166-kb plasmid (pShT-P). The sequence will represent the essential data for future phylogenetic and comparative genome studies of mycolactone-producing mycobacteria. PMID:27688344

  5. Mycobacterium avium subsp. paratuberculosis infection, immunology and pathology of livestock

    Science.gov (United States)

    Mycobacterium avium subsp. paratuberculosis (MAP) infection in ruminants leads to a chronic and progressive enteric disease (Johne’s disease) that results in loss of intestinal function, poor body condition, and eventual death. Transmission is primarily through a fecal-oral route in neonates but con...

  6. Autophagy modulates the Mycobacterium tuberculosis-induced cytokine response

    NARCIS (Netherlands)

    Kleinnijenhuis, J.; Oosting, M.; Plantinga, T.S.; Meer, J.W.M. van der; Joosten, L.A.B.; Crevel, R. van; Netea, M.G.

    2011-01-01

    Both autophagy and pro-inflammatory cytokines are involved in the host defence against mycobacteria, but little is known regarding the effect of autophagy on Mycobacterium tuberculosis (MTB)-induced cytokine production. In the present study, we assessed the effect of autophagy on production of monoc

  7. Mycobacterium tuberculosis Can Utilize Heme as an Iron Source▿

    OpenAIRE

    Jones, Christopher M.; Niederweis, Michael

    2011-01-01

    Most iron in mammals is found within the heme prosthetic group. Consequently, many bacterial pathogens possess heme acquisition systems to utilize iron from the host. Here, we demonstrate that Mycobacterium tuberculosis can utilize heme as an iron source, suggesting that M. tuberculosis possesses a yet-unknown heme acquisition system.

  8. Complete Genome Sequence of Mycobacterium ulcerans subsp. shinshuense.

    Science.gov (United States)

    Yoshida, Mitsunori; Nakanaga, Kazue; Ogura, Yoshitoshi; Toyoda, Atsushi; Ooka, Tadasuke; Kazumi, Yuko; Mitarai, Satoshi; Ishii, Norihisa; Hayashi, Tetsuya; Hoshino, Yoshihiko

    2016-01-01

    Mycobacterium ulcerans subsp. shinshuense produces mycolactone and causes Buruli ulcer. Here, we report the complete sequence of its genome, which comprises a 5.9-Mb chromosome and a 166-kb plasmid (pShT-P). The sequence will represent the essential data for future phylogenetic and comparative genome studies of mycolactone-producing mycobacteria. PMID:27688344

  9. Adaptation and evolution of drug-resistant Mycobacterium tuberculosis

    NARCIS (Netherlands)

    I.L. Bergval

    2013-01-01

    Many studies have been conducted on drug resistance and the evolution of Mycobacterium tuberculosis. Notwithstanding, many molecular mechanisms facilitating the emergence, adaptation and spread of drug-resistant tuberculosis have yet to be discovered. This thesis reports studies of the adaptive mech

  10. Variable host-pathogen compatibility in Mycobacterium tuberculosis.

    NARCIS (Netherlands)

    Gagneux, Sebastien; DeRiemer, Kathryn; Van, Tran; Kato-Maeda, Midori; Jong, Bouke C de; Narayanan, Sujatha; Nicol, Mark; Niemann, Stefan; Kremer, Kristin; Gutierrez, M Cristina; Hilty, Markus; Hopewell, Philip C; Small, Peter M

    2006-01-01

    Mycobacterium tuberculosis remains a major cause of morbidity and mortality worldwide. Studies have reported human pathogens to have geographically structured population genetics, some of which have been linked to ancient human migrations. However, no study has addressed the potential evolutionary c

  11. Human Mycobacterium bovis Infections in London and Southeast England

    OpenAIRE

    Stone, M J; Brown, T. J.; Drobniewski, F. A.

    2012-01-01

    Variable-number tandem repeat (VNTR) and spoligotyping analyses were used to assess transmission of Mycobacterium bovis between humans. VNTR was more discriminatory than spoligotyping. Low case numbers, despite a substantial animal reservoir, and resolution of all isolates provided no evidence of recent human-to-human transmission or recent significant infection from animals.

  12. Syntrophic biodegradation of butachlor by Mycobacterium sp. J7A and Sphingobium sp. J7B isolated from rice paddy soil.

    Science.gov (United States)

    Kim, Nam Hyun; Kim, Dong-Uk; Kim, Ijung; Ka, Jong-Ok

    2013-07-01

    Two bacterial strains involved in syntrophic degradation of chloroacetamide herbicide butachlor were isolated from a rice paddy soil. Analysis of 16S rRNA gene sequences indicated that the two isolates were related to members of the genera Mycobacterium and Sphingobium, respectively. Thus, a pair consisted of Mycobacterium sp. J7A and Sphingobium sp. J7B could rapidly degrade butachlor (100 mg L(-1)) at 28 °C within 24 h, while each isolate alone was not able to completely degrade butachlor. The isolate Mycobacterium sp. J7A was observed to grow slightly on butachlor, possibly utilizing the alkyl side chain of butachlor as its carbon and energy source, but the isolate Sphingobium sp. J7B alone could not grow on butachlor at all. Gas chromatography-mass spectrometry on catabolic intermediates revealed that the strain J7A produced and accumulated 2-chloro-N-(2,6-diethylphenyl) acetamide (CDEPA) during growth on butachlor. This intermediate was not further degraded by strain J7A, but strain J7B was observed to be able to completely degrade and grow on it through 2,6-diethylaniline (DEA). The results showed that butachlor was completely degraded by the two isolates by syntrophic metabolism, in which strain Mycobacterium sp. J7A degraded butachlor to CDEPA, which was subsequently degraded by strain Sphingobium sp. J7B through DEA.

  13. Business management replication from established market to emerging market: An analysis of the Indian Premier League from the Bangladesh market perspective

    Directory of Open Access Journals (Sweden)

    Shams, S M Riad

    2012-04-01

    Full Text Available This paper attempts to analyze the critical factors that would be crucial to replicating business management strategies from established market to emerging market. A case study is conducted on the Indian Premier League market and Bangladesh market, while these markets are considered as established and emerging markets of the Twenty20 cricket business consecutively. Seven critical factors are identified from the study as crucial in case of replicating business management strategies from established market to emerging market. However, the implementation of the critical factors should follow only the given situation, underlying the environment of the emerging market. The findings of the study shall shed light to managerial perception from the perspective of emerging market business strategies, while the strategies are learnt from another market.

  14. Massive gene acquisitions in Mycobacterium indicus pranii provide a perspective on mycobacterial evolution.

    Science.gov (United States)

    Saini, Vikram; Raghuvanshi, Saurabh; Khurana, Jitendra P; Ahmed, Niyaz; Hasnain, Seyed E; Tyagi, Akhilesh K; Tyagi, Anil K

    2012-11-01

    Understanding the evolutionary and genomic mechanisms responsible for turning the soil-derived saprophytic mycobacteria into lethal intracellular pathogens is a critical step towards the development of strategies for the control of mycobacterial diseases. In this context, Mycobacterium indicus pranii (MIP) is of specific interest because of its unique immunological and evolutionary significance. Evolutionarily, it is the progenitor of opportunistic pathogens belonging to M. avium complex and is endowed with features that place it between saprophytic and pathogenic species. Herein, we have sequenced the complete MIP genome to understand its unique life style, basis of immunomodulation and habitat diversification in mycobacteria. As a case of massive gene acquisitions, 50.5% of MIP open reading frames (ORFs) are laterally acquired. We show, for the first time for Mycobacterium, that MIP genome has mosaic architecture. These gene acquisitions have led to the enrichment of selected gene families critical to MIP physiology. Comparative genomic analysis indicates a higher antigenic potential of MIP imparting it a unique ability for immunomodulation. Besides, it also suggests an important role of genomic fluidity in habitat diversification within mycobacteria and provides a unique view of evolutionary divergence and putative bottlenecks that might have eventually led to intracellular survival and pathogenic attributes in mycobacteria. PMID:22965120

  15. Insight into the evolution and origin of leprosy bacilli from the genome sequence of Mycobacterium lepromatosis

    Science.gov (United States)

    Singh, Pushpendra; Benjak, Andrej; Schuenemann, Verena J.; Herbig, Alexander; Avanzi, Charlotte; Busso, Philippe; Nieselt, Kay; Krause, Johannes; Vera-Cabrera, Lucio; Cole, Stewart T.

    2015-01-01

    Mycobacterium lepromatosis is an uncultured human pathogen associated with diffuse lepromatous leprosy and a reactional state known as Lucio's phenomenon. By using deep sequencing with and without DNA enrichment, we obtained the near-complete genome sequence of M. lepromatosis present in a skin biopsy from a Mexican patient, and compared it with that of Mycobacterium leprae, which has undergone extensive reductive evolution. The genomes display extensive synteny and are similar in size (∼3.27 Mb). Protein-coding genes share 93% nucleotide sequence identity, whereas pseudogenes are only 82% identical. The events that led to pseudogenization of 50% of the genome likely occurred before divergence from their most recent common ancestor (MRCA), and both M. lepromatosis and M. leprae have since accumulated new pseudogenes or acquired specific deletions. Functional comparisons suggest that M. lepromatosis has lost several enzymes required for amino acid synthesis whereas M. leprae has a defective heme pathway. M. lepromatosis has retained all functions required to infect the Schwann cells of the peripheral nervous system and therefore may also be neuropathogenic. A phylogeographic survey of 227 leprosy biopsies by differential PCR revealed that 221 contained M. leprae whereas only six, all from Mexico, harbored M. lepromatosis. Phylogenetic comparisons indicate that M. lepromatosis is closer than M. leprae to the MRCA, and a Bayesian dating analysis suggests that they diverged from their MRCA approximately 13.9 Mya. Thus, despite their ancient separation, the two leprosy bacilli are remarkably conserved and still cause similar pathologic conditions. PMID:25831531

  16. Detection of Mycobacterium avium subsp. paratuberculosis in Cow Milk Using Culture and PCR methods

    Directory of Open Access Journals (Sweden)

    rezavand, B.,

    2011-12-01

    Full Text Available Mycobacterium avium subspecies paratuberculosis (MAP is the cause of John’s disease also calledparatuberculosis. This is economically one of the important infectious diseases in cattle and ruminanthusbandry. This disease is manifested as granulomatosis entrocolitis, lymphadenitis and inflammation locallymphatic vessels. The typical sign of this disease is progressive loss of weight. Considering the importanceof detection of this disease in this study, two methods, culture and PCR, were used for the identification ofthis microorganism. In this study 100 milk samples from apparently healthy cows and 100 milk samplesfrom cows that have been suspicious of John’s disease were taken from in Sarab, East Azarbaijan, Iran.Direct microscope observation after ziehl-neelsen staining was done. Then, bacterial culture on specificmedium was carried out, and finally, identification of Mycobacterium avium subsp. paratuberculosis wasexamined using PCR and specific primers. Using direct observation, culture and PCR analyses showed thatfrom 100 healthy cow milk samples, 8, 9 and 12 samples were positive MAP for each method respectively.The results of direct observation, culture and PCR analysis on affected cows were 15, 40 and 44, respectively. The results of this study showed that culture and PCR analyses methods are important in the identification of the causes of this disease. Therefore, considering the frequency of the disease in the studied region, either of those methods can be used in the microorganism identification.

  17. A new piperidinol derivative targeting mycolic acid transport in Mycobacterium abscessus.

    Science.gov (United States)

    Dupont, Christian; Viljoen, Albertus; Dubar, Faustine; Blaise, Mickaël; Bernut, Audrey; Pawlik, Alexandre; Bouchier, Christiane; Brosch, Roland; Guérardel, Yann; Lelièvre, Joël; Ballell, Lluis; Herrmann, Jean-Louis; Biot, Christophe; Kremer, Laurent

    2016-08-01

    The natural resistance of Mycobacterium abscessus to most commonly available antibiotics seriously limits chemotherapeutic treatment options, which is particularly challenging for cystic fibrosis patients infected with this rapid-growing mycobacterium. New drugs with novel molecular targets are urgently needed against this emerging pathogen. However, the discovery of such new chemotypes has not been appropriately performed. Here, we demonstrate the utility of a phenotypic screen for bactericidal compounds against M. abscessus using a library of compounds previously validated for activity against M. tuberculosis. We identified a new piperidinol-based molecule, PIPD1, exhibiting potent activity against clinical M. abscessus strains in vitro and in infected macrophages. Treatment of infected zebrafish with PIPD1 correlated with increased embryo survival and decreased bacterial burden. Whole genome analysis of M. abscessus strains resistant to PIPD1 identified several mutations in MAB_4508, encoding a protein homologous to MmpL3. Biochemical analyses demonstrated that while de novo mycolic acid synthesis was unaffected, PIPD1 strongly inhibited the transport of trehalose monomycolate, thereby abrogating mycolylation of arabinogalactan. Mapping the mutations conferring resistance to PIPD1 on a MAB_4508 tridimensional homology model defined a potential PIPD1-binding pocket. Our data emphasize a yet unexploited chemical structure class against M. abscessus infections with promising translational development possibilities. PMID:27121350

  18. Genotype heterogeneity of Mycobacterium tuberculosis within geospatial hotspots suggests foci of imported infection in Sydney, Australia.

    Science.gov (United States)

    Gurjav, Ulziijargal; Jelfs, Peter; Hill-Cawthorne, Grant A; Marais, Ben J; Sintchenko, Vitali

    2016-06-01

    In recent years the State of New South Wales (NSW), Australia, has maintained a low tuberculosis incidence rate with little evidence of local transmission. Nearly 90% of notified tuberculosis cases occurred in people born in tuberculosis-endemic countries. We analyzed geographic, epidemiological and genotypic data of all culture-confirmed tuberculosis cases to identify the bacterial and demographic determinants of tuberculosis hotspot areas in NSW. Standard 24-loci mycobacterium interspersed repetitive unit-variable number tandem repeat (MIRU-24) typing was performed on all isolates recovered between 2009 and 2013. In total 1692/1841 (91.9%) cases with confirmed Mycobacterium tuberculosis infection had complete MIRU-24 and demographic data and were included in the study. Despite some year-to-year variability, spatio-temporal analysis identified four tuberculosis hotspots. The incidence rate and the relative risk of tuberculosis in these hotspots were 2- to 10-fold and 4- to 8-fold higher than the state average, respectively. MIRU-24 profiles of M. tuberculosis isolates associated with these hotspots revealed high levels of heterogeneity. This suggests that these spatio-temporal hotspots, within this low incidence setting, can represent areas of predominantly imported infection rather than clusters of cases due to local transmission. These findings provide important epidemiological insight and demonstrate the value of combining tuberculosis genotyping and spatiotemporal data to guide better-targeted public health interventions. PMID:26187743

  19. Anti-Inflammatory and Antiapoptotic Responses to Infection: A Common Denominator of Human and Bovine Macrophages Infected with Mycobacterium avium Subsp. paratuberculosis

    Directory of Open Access Journals (Sweden)

    Naiara Abendaño

    2013-01-01

    Full Text Available Mycobacterium avium subsp. paratuberculosis (Map is the causative agent of a chronic intestinal inflammation in ruminants named Johne's disease or paratuberculosis and a possible etiopathological agent of human Crohn's disease (CD. Analysis of macrophage transcriptomes in response to Map infection is expected to provide key missing information in the understanding of the role of this pathogen in establishing an inappropriate and persistent infection in a susceptible host and of the molecular mechanisms that might underlie the early phases of CD. In this paper we summarize transcriptomic studies of human and bovine peripheral blood mononuclear cells (PBMC, monocyte-derived macrophages (MDMs, and macrophages-like cell lines in vitro infected with Map. Most studies included in this paper consistently reported common gene expression signatures of bovine and human macrophages in response to Map such as enhanced expression of the anti-inflammatory cytokines IL-10 and IL-6, which promote bacterial survival. Overexpression of IL-10 could be responsible for the Map-associated reduction in the expression of the proapoptotic TNF-α gene observed in bovine and human macrophages.

  20. Biodegradation of 2-ethylhexyl nitrate by Mycobacterium austroafricanum IFP 2173

    International Nuclear Information System (INIS)

    Compounds such as 2-ethylhexyl nitrate (2-EHN) are added to diesel fuel to improve ignition and boost cetane number. The production of 2-EHN reaches around 100000 tons per year in France, principally. Risks associated to its utilization are however poorly known because, in case of accidental release in the environment, nothing is known about its biodegradation. In this study, we aimed at (i) identifying bacterial strains able to degrade 2-EHN and compare their capabilities, (ii) elucidating the degradation pathway, and (iii) identifying the enzymes involved. Biodegradation of 2-EHN was first tested in biphasic cultures under conditions that reduce the toxicity and increase the availability of the hydrophobic substrate. Using optimized culture conditions, we showed that several strains of Mycobacterium austroafricanum were able to degrade 2-EHN. One of the most efficient strain (IFP 2173) which could grow at 2-EHN concentrations up to 6 g.L-1, was chosen to investigate the degradation pathway. On the basis of carbon balance determination and gas chromatographic (GC) analysis on the culture medium, I found that the degradation of 2-EHN was incomplete and gave rise to the accumulation of a metabolite. This metabolite was identified as β-methyl-γ-butyrolactone by GC-MS and LC-MS/MS analysis. The structure of the lactone indicated that 2-EHN was degraded through a pathway involving the hydroxylation of the methyl group of the main carbon chain, its oxidation into aldehyde an acid and a subsequent cycle of b-oxidation. Enzymes involved in the 2-EHN biodegradation pathway were looked for by a proteomic approach. Analyses by two-dimensional gel electrophoresis showed that, when exposed to 2-EHN, strain IFP 2173 triggered the synthesis of a bunch of enzymes specialized in fatty acid metabolism such as β-oxidation enzymes, as well as alcohol and aldehyde dehydrogenases. An exhaustive analysis of the IFP 2173 proteome resulted in the identification of more than 200

  1. Dual channel real-time PCR melting curve analysis-based assay for detecting of anti-tuberculosis drug-resistant mutations in Mycobacterium tuberculosis%双通道实时荧光PCR熔解曲线法检测结核分枝杆菌药物耐药相关基因突变

    Institute of Scientific and Technical Information of China (English)

    柳清云; 罗涛; 李静; 梅建; 高谦

    2013-01-01

    目的 基于双标记荧光探针熔解曲线分析技术,建立一种利用实时荧光PCR快速检测结核分枝杆菌耐药突变的方法.方法 根据结核分枝杆菌一线药物常见耐药突变位点(包括rpoB 81 bp耐药决定区、inhA启动子、katG315、ahpC启动子以及embB306)设计6条荧光双标记探针和对应引物,通过PCR扩增耐药突变位点所在基因片段,在扩增完成后通过熔解曲线检测分析实现对耐药突变的快速检测.通过对2008年上海市疾控中心收集的76株临床耐多药(MDR)菌株进行检测,验证本方法的敏感度和特异度.结果 本方法成功从76株MDR菌株中检测出相关耐药突变,各种突变对应ATm值范围为1.8~14.4℃.将检测结果和测序结果对比表明该方法检测敏感度和特异度都为100%(rpoB,80/80:inhA,7/7;katG315,59/59;ahpC,8/8;embB306,27/27).本方法可以成功从最低浓度为100拷贝/μl的结核分枝杆菌DNA样本中准确地检测耐药突变.结论 双通道实时荧光PCR熔解曲线法可以快速灵敏地检测结核分枝杆菌常见耐药突变.该方法具有检测迅速准确、结果易判读、交叉污染概率低等特点,可用于快速检测临床结核耐药相关的基因突变,并对结核耐药情况进行评估.%Objective Based on dual channel melting curve analysis-based assay,we developed a method to rapidly detect the drug-resistant mutations in Mycobacterium tuberculosis through real-time PCR.Methods According to the common first-line drug-resistant mutations of Mycobacterium tuberculosis,we designed six dual-labeled fluorescence probes to rapidly detect the drug-resistant mutations through realtime PCR melting curve after amplifications of drug-resistant related gene region of DNA.The targets include rpoB 81 bp core region,katG315,inhA promoter,ahpC promoter and embB306.To validate the sensitivity and specificity of our method,we performed real-time PCR assays to detect drug-resistant mutations in 76

  2. Infections with Mycobacterium tuberculosis and Mycobacterium avium among HIV-infected patients after the introduction of highly active antiretroviral therapy. EuroSIDA Study Group JD

    DEFF Research Database (Denmark)

    Kirk, O; Gatell, J M; Mocroft, A;

    2000-01-01

    the introduction of HAART, using data from the EuroSIDA study, a European, multicenter observational cohort of more than 7,000 patients. Overall incidences of Mycobacterium tuberculosis (TB) and Mycobacterium avium complex (MAC) were 0.8 and 1.4 cases/100 person-years of follow-up (PYF), decreasing from 1.8 (TB...

  3. The Analysis of the Relationship between Organizational Structure and Information Technology (IT): And the Barriers to Its Establishment at the University of Isfahan from the Faculty Member's Viewpoints

    Science.gov (United States)

    Peyman, Yarmohammadzadeh; Mohsen, Allammeh Sayyed; Hassan, Ghalavandi; Aboulghassim, Farhang; Zaman, Ajdari

    2011-01-01

    The purpose of the present study is to investigate the relationship between organizational structure between IT and the barriers to its establishment in University of Isfahan from faculty member's viewpoints in 2007-2008. The questionnaires were prepared and examined based on the organization dimensions of organizational structures (formality,…

  4. Analysis on the Influencing Factors of the Quality Traceable System Established by Edible Agricultural Products Enterprises-Taking Sichuan as an Example

    Institute of Scientific and Technical Information of China (English)

    2011-01-01

    According to the investigation data from 81 edible agricultural products enterprises in Sichuan Province,the influence factors of establishing quality tracing system are empirically analyzed from four aspects,including the enterprise features,the attitudes of operators,management and market competition by using the Logistic model.The results show that the enterprise can establish the quality tracing system after the comprehensive function of a series of internal and external factors.The four factors include peer pressure;age of enterprise;export and the expectation on improving economic interests have the biggest impact on enterprise to establish the quality tracing system.The factors,including the quality safety certificate obtained by the enterprises,export of products,sampling frequency,peer pressure,the pressure from consumers and media,the expectation on improving the competition of products,the expectation on improving economic interests,play a promotion role in helping enterprises to establish quality tracing system.The countermeasures and suggestion are put forward from strengthening the social responsibility of enterprises;intensifying the law enforcement of government and expanding the experimental enterprises with the quality tracing system.

  5. 瓜实蝇DNA甲基化的MSAP体系建立与优化%Establishment and optimization of MSAP Analysis System for DNA Methylation in Bactrocera cucurbitae

    Institute of Scientific and Technical Information of China (English)

    龚治; 周世豪; 马华博; 张亚楠; 符悦冠

    2016-01-01

    The melon fly [Bactrocera cucurbitae (Coquillett)] is one of the most important vegetables insect pest in China. However, the DNA methylation in melon fly has not been reported yet. Methylation sensitive amplified polymorphism (MSAP) is an important research tecnnique to analysis the DNA methylation. The MSAP can be established through the enzyme digestion reaction, ligation, PCR amplification system, and primer screening, such as 10 U restriction endonuclease is put in the 20 μL enzyme digestion system, and then react with 600 ng of genome DNA at 37℃ overnight; 1U of T4 ligase is put in the 20 μL ligation system, 50 pmol of HpaⅡ-MspⅠ-adapter and 5 pmol of EcoR I-adapter which react at 16℃ for 12 hours. Diluent ligation products were used in PCR to pre-amplification and selected amplification, the result is tested by silver staining and 6% denaturing polyacrylamide gel electrophoresis. Six pairs of primers which suited for DNA methylation sensitive amplified polymorphism were selected out by this system. The MSAP system provides the technical support for the study of the epigenetics research of melon fly.%瓜实蝇[Bactrocera cucurbitae (Coquillett)]是中国重要的蔬菜害虫,但其DNA甲基化研究尚未见报道。甲基化敏感扩增多态性是研究DNA甲基化的重要技术之一。通过对酶切反应、连接、 PCR扩增和引物筛选等条件优化,建立瓜实蝇MSAP反应体系,即:①20μL酶切体系中加入10 U的限制性内切酶与600 ng基因组DNA,于37℃酶切反应过夜;②20μL连接体系中加入T4连接酶1 U, HpaⅡ-MspⅠ-adapter接头50 pmol, EcoR I-adapter接头5 pmol,并于16℃反应12 h;③连接产物稀释后进行PCR预扩增和选择性扩增,再经6%变性聚丙烯酰胺凝胶电泳和银染检测结果。通过该体系筛选出适用于瓜实蝇基因组DNA甲基化多态性研究的6对引物;瓜实蝇MSAP体系为瓜实蝇的表观遗传学研究提供了技术支持。

  6. Flow cytometry as a tool to identify Mycobacterium tuberculosis interaction with the immune system and drug susceptibility

    Directory of Open Access Journals (Sweden)

    Maria da Gloria Bonecini-Almeida

    2000-08-01

    Full Text Available Flow cytometric analysis is a useful and widely employed tool to identify immunological alterations caused by different microorganisms, including Mycobacterium tuberculosis. However, this tool can be used for several others analysis. We will discuss some applications for flow cytometry to the study of M. tuberculosis, mainly on cell surface antigens, mycobacterial secreted proteins, their interaction with the immune system using inflammatory cells recovered from peripheral blood, alveolar and pleura spaces and the influence of M. tuberculosis on apoptosis, and finally the rapid determination of drug susceptibility. All of these examples highlight the usefulness of flow cytometry in the study of M. tuber-culosis infection.

  7. Development of a new, combined rapid method using phage and PCR for detection and identification of viable Mycobacterium paratuberculosis bacteria within 48 hours.

    Science.gov (United States)

    Stanley, Emma C; Mole, Richard J; Smith, Rebecca J; Glenn, Sarah M; Barer, Michael R; McGowan, Michael; Rees, Catherine E D

    2007-03-01

    The FASTPlaqueTB assay is an established diagnostic aid for the rapid detection of Mycobacterium tuberculosis from human sputum samples. Using the FASTPlaqueTB assay reagents, viable Mycobacterium avium subsp. paratuberculosis cells were detected as phage plaques in just 24 h. The bacteriophage used does not infect M. avium subsp. paratuberculosis alone, so to add specificity to this assay, a PCR-based identification method was introduced to amplify M. avium subsp. paratuberculosis-specific sequences from the DNA of the mycobacterial cell detected by the phage. To give further diagnostic information, a multiplex PCR method was developed to allow simultaneous amplification of either M. avium subsp. paratuberculosis or M. tuberculosis complex-specific sequences from plaque samples. Combining the plaque PCR technique with the phage-based detection assay allowed the rapid and specific detection of viable M. avium subsp. paratuberculosis in milk samples in just 48 h.

  8. Characterization of the first report of Mycobacterium timonense infecting an HIV patient in an Ecuadorian hospital.

    Science.gov (United States)

    Zurita, J; Ortega-Paredes, D; Mora, M; Espinel, N; Parra, H; Febres, L; Zurita-Salinas, C

    2014-12-01

    Mycobacterium timonense is a non-tuberculous mycobacteria (NTM) described in southern France in 2009, and to our knowledge, not reported again as a human pathogen in indexed literature. The aim of this work was to characterize the first clinical isolate of M. timonense in Ecuador. Time of growth, biochemical tests, thin layer growth test, PCR-RFLP analysis of the hsp65 gene and MALDI-TOF spectra analysis were not able to identify the species. The species identification was achieved through sequencing of rrs, hsp65 and rpoB genes. The results highlight the necessity to set up a sequencing method to identify emerging NTM in Ecuadorian clinical facilities. PMID:24813256

  9. Segmentation of touching mycobacterium tuberculosis from Ziehl-Neelsen stained sputum smear images

    Science.gov (United States)

    Xu, Chao; Zhou, Dongxiang; Liu, Yunhui

    2015-12-01

    Touching Mycobacterium tuberculosis objects in the Ziehl-Neelsen stained sputum smear images present different shapes and invisible boundaries in the adhesion areas, which increases the difficulty in objects recognition and counting. In this paper, we present a segmentation method of combining the hierarchy tree analysis with gradient vector flow snake to address this problem. The skeletons of the objects are used for structure analysis based on the hierarchy tree. The gradient vector flow snake is used to estimate the object edge. Experimental results show that the single objects composing the touching objects are successfully segmented by the proposed method. This work will improve the accuracy and practicability of the computer-aided diagnosis of tuberculosis.

  10. Proteomic profiling of Mycobacterium tuberculosis identifies nutrient-starvation-responsive toxin-antitoxin systems

    DEFF Research Database (Denmark)

    Albrethsen, Jakob; Agner, Jeppe; Piersma, Sander R;

    2013-01-01

    In order to successfully enter the latent stage, Mycobacterium tuberculosis must adapt to conditions such as nutrient limitation and hypoxia. In vitro models that mimic latent infection are valuable tools for describing the changes in metabolism that occur when the bacterium exists in a non......-growing form. We used two complementary proteomic approaches, label-free LC-MS/MS analysis and two-dimensional difference gel electrophoresis, to determine the proteome profile of extracellular proteins from M. tuberculosis cultured under nutrient starvation. Through the label-free LC-MS/MS analysis......, significant differences in the overall metabolism during nutrient starvation were detected. Notably, members of the toxin-antitoxin systems were present in larger quantities in nutrient-starved cultures, supporting a role for these global modules as M. tuberculosis switches its metabolism into dormancy...

  11. Secular Religious Establishment

    DEFF Research Database (Denmark)

    Lægaard, Sune

    2013-01-01

    Secularism as a political doctrine claims that religion and politics should be separated. The compatibility question is whether secularism can accept some forms of religious establishment in the form of institutional linkages between state and organised religion. I argue that the answer to the co...

  12. Mycobacterium avium Subsp. avium Infection in Four Veal Calves: Differentiation from Intestinal Tuberculosis

    OpenAIRE

    2014-01-01

    Mycobacterium avium subsp. avium (Maa) is an intracellular pathogen belonging to the Mycobacterium avium-intracellulare complex (MAC). Reservoirs of MAC are the natural environment, wildlife and domestic animals. In adult bovine, MAC infections are typically caused by Mycobacterium avium subsp. paratuberculosis (Map). Maa infections in bovine are rarely reported but may cause clinical disease and pathological lesions similar to those observed in paratuberculosis or those induced by members of...

  13. High-catalase strains of Mycobacterium kansasii isolated from water in Texas.

    OpenAIRE

    Steadham, J E

    1980-01-01

    Isolation techniques with membrane-filtered potable water samples resulted in the isolation of potentially pathogenic high-catalase strains of Mycobacterium kansasii from 8 of 19 representative outlets in a small central Texas town. Mycobacterium gordonae was isolated from all samples, and Mycobacterium fortuitum was isolated from two samples. Data on chlorine levels are presented along with a possible explanation for the unusually high numbers of mycobacteria in these potable water samples. ...

  14. Relationships between Mycobacterium Isolates from Patients with Pulmonary Mycobacterial Infection and Potting Soils▿ †

    OpenAIRE

    De Groote, Mary Ann; Pace, Norman R.; Fulton, Kayte; Joseph O. Falkinham

    2006-01-01

    High numbers of mycobacteria, including known pathogenic species such as Mycobacterium avium, Mycobacterium intracellulare, and Mycobacterium chelonae, were recovered from aerosols produced by pouring commercial potting soil products and potting soil samples provided by patients with pulmonary mycobacterial infections. The dominant mycobacteria in the soil samples corresponded to the dominant species implicated clinically. Profiles of large restriction fragments obtained by pulsed-field gel e...

  15. Comparative Ser/Thr/Tyr phosphoproteomics between two Mycobacterial species: The fast growing Mycobacterium smegmatis and the slow growing Mycobacterium bovis BCG.

    Directory of Open Access Journals (Sweden)

    Kehilwe Confidence Nakedi

    2015-04-01

    Full Text Available Ser/Thr/Tyr protein phosphorylation plays a critical role in regulating mycobacterial growth and development. Understanding the mechanistic link between protein phosphorylation signaling network and mycobacterial growth rate requires a global view of the phosphorylation events taking place at a given time under defined conditions. In the present study we employed a phosphopeptide enrichment and high throughput mass spectrometry-based strategy to investigate and qualitatively compare the phosphoproteome of two mycobacterial model organisms: the fast growing Mycobacterium smegmatis and the slow growing Mycobacterium bovis BCG. Cells were harvested during exponential phase and our analysis detected a total of 185 phospho-sites in M. smegmatis, of which 106 were confidently localized (localization probability (LP =0.75; PEP=0.01. By contrast, in M. bovis BCG the phosphoproteome comprised 442 phospho-sites, of which 289 were confidently localized. The percentage distribution of Ser/Thr/Tyr phosphorylation was 39.47, 57.02 and 3.51 % for M. smegmatis and 35, 61.6 and 3.1% for M. bovis BCG. Moreover, our study identified a number of conserved Ser/Thr phosphorylated sites and conserved Tyr phosphorylated sites across different mycobacterial species. Overall a qualitative comparison of the fast and slow growing mycobacteria suggests that the phosphoproteome of M. smegmatis is a simpler version of that of M. bovis BCG. In particular, M. bovis BCG exponential cells exhibited a much more complex and sophisticated protein phosphorylation network regulating important cellular cycle events such as cell wall biosynthesis, elongation, cell division including immediately response to stress. The differences in the two phosphoproteomes are discussed in light of different mycobacterial growth rates.

  16. Multidrug-Resistant Mycobacterium tuberculosis of the Latin American Mediterranean Lineage, Wrongly Identified as Mycobacterium pinnipedii (Spoligotype International Type 863 [SIT863]), Causing Active Tuberculosis in South Brazil

    KAUST Repository

    Dalla Costa, Elis R.

    2015-09-23

    We recently detected the spoligotype patterns of strains of Mycobacterium pinnipedii, a species of the Mycobacterium tuberculosis complex, in sputum samples from nine cases with pulmonary tuberculosis residing in Porto Alegre, South Brazil. Because this species is rarely encountered in humans, we further characterized these nine isolates by additional genotyping techniques, including 24-locus mycobacterial interspersed repetitive-unit–variable-number tandem-repeat (MIRU-VNTR) typing, verification of the loci TbD1, RD9, pks15/1, RDRio, and fbpC, the insertion of IS6110 at a site specific to the M. tuberculosis Latin American Mediterranean (LAM) lineage, and whole-genome sequencing. The combined analysis of these markers revealed that the isolates are in fact M. tuberculosis and more specifically belong to the LAM genotype. Most of these isolates (n = 8) were shown to be multidrug resistant (MDR), which prompted us to perform partial sequencing of the rpoA, rpoB, rpoC, katG, and inhA genes. Seven isolates (77.8%) carried the S315T mutation in katG, and one of these (11%) also presented the C(−17)T single-nucleotide polymorphism (SNP) in inhA. Interestingly, six of the MDR isolates also presented an undescribed insertion of 12 nucleotides (CCA GAA CAA CCC) in codon 516 of rpoB. No putative compensatory mutation was found in either rpoA or rpoC. This is the first report of an M. tuberculosis LAM family strain with a convergent M. pinnipedii spoligotype. These spoligotypes are observed in genotype databases at a modest frequency, highlighting that care must be taken when identifying isolates in the M. tuberculosis complex on the basis of single genetic markers.

  17. Pneumopatia causada por Mycobacterium kansasii Lung disease caused by Mycobacterium kansasii

    Directory of Open Access Journals (Sweden)

    Nelson Morrone

    2003-12-01

    Full Text Available INTRODUÇÃO: O Mycobacterium kansasii é uma micobactéria não tuberculosa que pode causar colonização ou infecção pulmonar. OBJETIVO: Relatar experiência com doença pulmonar causada pelo M. kansasii em uma série de seis pacientes diagnosticados ao longo de cinco anos. MÉTODO: Entre junho de 1995 e junho de 2000 foram admitidos 1.349 pacientes no Dispensário do Ipiranga Ari Nogueira da Silva-Sanatorinhos, com o diagnóstico de tuberculose pulmonar, dos quais seis tiveram cultura positiva para M. kansasii. RESULTADOS: Cinco pacientes eram homens e a idade variou entre 25 e 77 anos. Todos apresentavam pneumopatia crônica e eram sintomáticos respiratórios com teste negativo para síndrome de imunodeficiência humana. As radiografias de tórax eram compatíveis com a presença de doença pulmonar prévia: cavidades de paredes finas foram notadas em todos e espessamento pleural subjacente às cavidades foi observado em dois pacientes. Todos foram tratados inicialmente com isoniazida, rifampicina, pirazinamida (INH-RMP-PZA e etambutol (EMB foi introduzido precocemente em dois pacientes por intolerância à pirazinamida, enquanto que em outros dois a introdução foi feita ao ser conhecido o resultado da cultura. Todos os pacientes foram tratados por mais de nove meses, tendo sido observada recidiva em um deles. Um paciente com silicose faleceu após dois anos por insuficiência respiratória, depois de ter sido considerado curado. CONCLUSÕES: A micobacteriose por M. kansasii foi encontrada com baixa freqüência, podendo estar relacionada às características dos pacientes encaminhados ao nosso serviço. O esquema INH-RMP-PZA, com substituição eventual da PZA por etambutol, mostrou sucesso terapêutico.BACKGROUND: Mycobacterium kansasii is a nontuberculous mycobacterium that can colonize the lungs and cause pulmonary infection. OBJECTIVE: To report authors' study of 6 patients with pulmonary disease caused by M. kansasii infection in

  18. Xpert MTB/RIF test for rapid diagnosis of Mycobacterium tuberculosis and simultaneous detection of multidrugresistant tuberculous bacillus

    Directory of Open Access Journals (Sweden)

    Chuang CAI

    2012-10-01

    Full Text Available Tuberculosis (TB is endemic in China with high prevalence of multiple-drug resistant tuberculous bacilli (MDRTB. The incidence of new cases of TB reaches 1,300 thousand annually. Among them, 5.7 percent are MDR-TB. Staining for acidfast bacilli in sputum and clinicoradiological examination have been the main diagnostic tools for TB, particularly pulmonary TB. However, the positive rate of sputum Ziehl-Neelsen stain for sputum is disappointedly low, merely 28% in newly-diagnosed TB. Moreover, the radiological manifestations of the patients suspected of TB are often non-specific. All these facts call for a simple, accurate and rapid diagnostic method to overcome this bottleneck, which hinders the success of satisfactory TB control in China. Employing both hemi-nested RT-PCR and beacon technology with fluorescent probes, the MTB/RIF diagnostic assay specifically amplifies, thus helps detect the rpoB gene, which is unique to Mycobacterium tuberculosis and also a biomolecularmarker of rifampin resistance. As a semi-quantitative analysis, the quantity of Mycobacterium tuberculosis in samples is reflected by the threshold of PCR cycles during MTB/RIF assay. With Mycobacterium tuberculosis culture as the standard reference, for sputum samples from patients suspected of suffering from pulmonary TB, overall diagnostic sensitivity of MTB/RIF assay is 73.1%–90.0% with a specificity of 99.0%–99.5%. For detection of rpoB gene mutations responsible for rifampin-resistance, the sensitivity is 97.2% and specificity is 98.3%. Following sample loading, the system can automatically complete the diagnostic process and report the results within 2 hours. Targeting the rpoB gene specifically, there is no cross-reaction with non-tuberculosis mycobacteria or other common respiratory pathogens. In addition to sputum samples, the system can be used to detect Mycobacterium tuberculosis in various body fluids (including pleural effusion, urine, cerebrospinal fluid and

  19. 结核病3种实验诊断方法比较与分枝杆菌耐药性分析%Comparison of three methods in the detection of tuberculosis and analysis of drug resistance in clinical isolates of Mycobacterium

    Institute of Scientific and Technical Information of China (English)

    陈建波; 黄同花; 陈郁筠; 吴驰; 肖颜玉; 罗凯; 叶飞娣

    2012-01-01

    Objective To compare of three methods in tuberculosis detection and analyze the drug resistance of Mycobacterium isolats. Methods The results of smear-microscopy, Mycobacterium culture, and PCR detection of patients with definite diagnosis of tuberculosis in Shenzhen Third People s Hospital between 2009 and 2010 were analyzed, including the species of Mycobacterium and drug resistance. Results Total 640 hospitalized patients with well medical records were enrolled in this study,including 537 cases of pulmonary tuberculosis, 82 of tuberculosis pleurisy and 21 of tuberculosis meningitis, the positive rates of smear-microscopy of whom were 44. 1% ,5. 3% and 13. 3% ,of Mycobacterium-culture were 59. 9% , 12. 9% and 30. 0% , and of PCR detection were 52. 2% , 16. 7% and 26. 3% respectively. Species of Mycobacterium were made up with 67. 0% of Mycobacterium tuberculosis, 16. 8% of Mycobacterium bovis and 16. 2% of Mycobacterium other than tuberculosis, which were confirmed by Mycobacterium isolates identification. The drug resistance of Mycobacterium tuberculosis to Rifampin, Isoniazid, Kthambutol and Streptomycin were 16. 7%, 15. 1%,4. 8% and 15. 9% respectively. Conclusion The main laboratory diagnostic methods of tuberculosis were smear-microscopy, Mycrobacterium-culture and PCR detection at present in Shenzhen, and most common pathogens that cause tuberculosis were Mycobacterium tuberculosis with high level drug resistance to three mainly used anti-tuberculosis drugs presently. In addition,about 40% tuberculosis patients were negative with Mycobacterium,bringing difficulties to identify active tuberculosis and old inactive tuberculosis, indicating that new and advanced diagnostic methods should be further researched.%目的 比较结核病3种实验诊断方法,分析目前深圳市分枝杆菌耐药情况.方法 统计2009~2010年深圳市第三人民医院临床确诊结核病患者痰标本涂片镜检、培养和PCR检测结果,并对分枝杆菌菌种及对

  20. Genetic diversity of Mycobacterium tuberculosis isolated from tuberculosis patients in the Serengeti ecosystem in Tanzania.

    Science.gov (United States)

    Mbugi, Erasto V; Katale, Bugwesa Z; Siame, Keith K; Keyyu, Julius D; Kendall, Sharon L; Dockrell, Hazel M; Streicher, Elizabeth M; Michel, Anita L; Rweyemamu, Mark M; Warren, Robin M; Matee, Mecky I; van Helden, Paul D

    2015-03-01

    This study was part of a larger cross-sectional survey that was evaluating tuberculosis (TB) infection in humans, livestock and wildlife in the Serengeti ecosystem in Tanzania. The study aimed at evaluating the genetic diversity of Mycobacterium tuberculosis isolates from TB patients attending health facilities in the Serengeti ecosystem. DNA was extracted from 214 sputum cultures obtained from consecutively enrolled newly diagnosed untreated TB patients aged ≥18 years. Spacer oligonucleotide typing (spoligotyping) and Mycobacterium Interspersed Repetitive Units and Variable Number Tandem Repeat (MIRU-VNTR) were used to genotype M. tuberculosis to establish the circulating lineages. Of the214 M. tuberculosis isolates genotyped, 55 (25.7%) belonged to the Central Asian (CAS) family, 52 (24.3%) were T family (an ill-defined family), 38 (17.8%) belonged to the Latin American Mediterranean (LAM) family, 25 (11.7%) to the East-African Indian (EAI) family, 25 (11.7%) comprised of different unassigned ('Serengeti') strain families, while 8 (3.7%) belonged to the Beijing family. A minority group that included Haarlem, X, U and S altogether accounted for 11 (5.2%) of all genotypes. MIRU-VNTR typing produced diverse patterns within and between families indicative of unlinked transmission chains. We conclude that, in the Serengeti ecosystem only a few successful families predominate namely CAS, T, LAM and EAI families. Other types found in lower prevalence are Beijing, Haarlem, X, S and MANU. The Haarlem, EAI_Somalia, LAM3 and S/convergent and X2 subfamilies found in this study were not reported in previous studies in Tanzania.