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Sample records for leprae reca intein

  1. Dynamics differentiate between active and inactive inteins.

    Science.gov (United States)

    Cronin, Melissa; Coolbaugh, Michael J; Nellis, David; Zhu, Jianwei; Wood, David W; Nussinov, Ruth; Ma, Buyong

    2015-02-16

    The balance between stability and dynamics for active enzymes can be somewhat quantified by studies of intein splicing and cleaving reactions. Inteins catalyze the ligation of flanking host exteins while excising themselves. The potential for applications led to engineering of a mini-intein splicing domain, where the homing endonuclease domain of the Mycobacterium tuberculosis RecA (Mtu recA) intein was removed. The remaining domains were linked by several short peptides, but splicing activity in all was substantially lower than the full-length intein. Native splicing activity was restored in some cases by a V67L mutation. Using computations and experiments, we examine the impact of this mutation on the stability and conformational dynamics of the mini-intein splicing domain. Molecular dynamics simulations were used to delineate the factors that determine the active state, including the V67L mini-intein mutant, and peptide linker. We found that (1) the V67L mutation lowers the global fluctuations in all modeled mini-inteins, stabilizing the mini-intein constructs; (2) the connecting linker length affects intein dynamics; and (3) the flexibilities of the linker and intein core are higher in the active structure. We have observed that the interaction of the linker region and a turn region around residues 35-41 provides the pathway for the allostery interaction. Our experiments reveal that intein catalysis is characterized by non-linear Arrhenius plot, confirming the significant contribution of protein conformational dynamics to intein function. We conclude that while the V67L mutation stabilizes the global structure, cooperative dynamics of all intein regions appear more important for intein function than high stability. Our studies suggest that effectively quenching the conformational dynamics of an intein through engineered allosteric interactions could deactivate intein splicing or cleaving. Copyright © 2014 Elsevier Masson SAS. All rights reserved.

  2. Exploring Intein Inhibition by Platinum Compounds as an Antimicrobial Strategy.

    Science.gov (United States)

    Chan, Hon; Pearson, C Seth; Green, Cathleen M; Li, Zhong; Zhang, Jing; Belfort, Georges; Shekhtman, Alex; Li, Hongmin; Belfort, Marlene

    2016-10-21

    Inteins, self-splicing protein elements, interrupt genes and proteins in many microbes, including the human pathogen Mycobacterium tuberculosis Using conserved catalytic nucleophiles at their N- and C-terminal splice junctions, inteins are able to excise out of precursor polypeptides. The splicing of the intein in the mycobacterial recombinase RecA is specifically inhibited by the widely used cancer therapeutic cisplatin, cis-[Pt(NH 3 ) 2 Cl 2 ], and this compound inhibits mycobacterial growth. Mass spectrometric and crystallographic studies of Pt(II) binding to the RecA intein revealed a complex in which two platinum atoms bind at N- and C-terminal catalytic cysteine residues. Kinetic analyses of NMR spectroscopic data support a two-step binding mechanism in which a Pt(II) first rapidly interacts reversibly at the N terminus followed by a slower, first order irreversible binding event involving both the N and C termini. Notably, the ligands of Pt(II) compounds that are required for chemotherapeutic efficacy and toxicity are no longer bound to the metal atom in the intein adduct. The lack of ammine ligands and need for phosphine represent a springboard for future design of platinum-based compounds targeting inteins. Because the intein splicing mechanism is conserved across a range of pathogenic microbes, developing these drugs could lead to novel, broad range antimicrobial agents. © 2016 by The American Society for Biochemistry and Molecular Biology, Inc.

  3. Lepra familiar

    Directory of Open Access Journals (Sweden)

    Richard Arrieta

    2001-09-01

    Full Text Available La lepra es entre cinco y diez veces más frecuente entre los convivientes de enfermos multibacilares. El objetivo de este trabajo es presentar el estudio de una mujer de 22 años con lepra lepromatosa, que condujo a la investigación de la enfermedad en sus hijos, sobrinos y familiares. Mediante el interrogatorio de la enferma, consulta médica de convivientes y visita domiciliaria, se pudieron demostrar nueve casos adicionales de lepra, por clínica e histopatología. Entre los diez niños habitantes de la misma casa, con edades entre los 3 meses y los 10 años, 7 tenían diversas formas de lepra. Una hermana de 30 años y un cuñado del caso índice, convivientes, tenían lepra indeterminada (LI. Los niños con lepra eran tres hijos de la primera paciente, su hermana de ocho años de edad y tres sobrinos, hijos de los padres con LI Cinco niños tenían lepra paucibacilar y dos, lepra multibacilar, dimoria lepromatosa; estos últimos, hijos de la madre lepromatosa. Todos son desplazados y viven en condiciones de hacinamiento y pobreza extremas. Presentaban signos de desnutrición moderada. Sólo uno de los niños tenía cicatriz de BCG. Los pacientes recibieron tratamiento con poliquimioterapia y los libres de lepra, vacunación con BCG. La fuente de contagio en una madre lepromatosa y las condiciones de pobreza extrema favorecieron la presentación de este brote intrafamiliar de lepra con serio compromiso de los niños. Todo diagnóstico de lepra debe llevar a la búsqueda de otros casos entre contactos y convivientes para lograr la detección precoz, el tratamiento oportuno y la prevención de discapacidades, objetivos básicos del Programa Nacional de Control de la Lepra.

  4. Protein trans-splicing of multiple atypical split inteins engineered from natural inteins.

    Directory of Open Access Journals (Sweden)

    Ying Lin

    Full Text Available Protein trans-splicing by split inteins has many uses in protein production and research. Splicing proteins with synthetic peptides, which employs atypical split inteins, is particularly useful for site-specific protein modifications and labeling, because the synthetic peptide can be made to contain a variety of unnatural amino acids and chemical modifications. For this purpose, atypical split inteins need to be engineered to have a small N-intein or C-intein fragment that can be more easily included in a synthetic peptide that also contains a small extein to be trans-spliced onto target proteins. Here we have successfully engineered multiple atypical split inteins capable of protein trans-splicing, by modifying and testing more than a dozen natural inteins. These included both S1 split inteins having a very small (11-12 aa N-intein fragment and S11 split inteins having a very small (6 aa C-intein fragment. Four of the new S1 and S11 split inteins showed high efficiencies (85-100% of protein trans-splicing both in E. coli cells and in vitro. Under in vitro conditions, they exhibited reaction rate constants ranging from ~1.7 × 10(-4 s(-1 to ~3.8 × 10(-4 s(-1, which are comparable to or higher than those of previously reported atypical split inteins. These findings should facilitate a more general use of trans-splicing between proteins and synthetic peptides, by expanding the availability of different atypical split inteins. They also have implications on understanding the structure-function relationship of atypical split inteins, particularly in terms of intein fragment complementation.

  5. PRP8 intein in cryptic species of Histoplasma capsulatum: evolution and phylogeny

    Science.gov (United States)

    The PRP8 intein is the most widespread intein among the Kingdom Fungi. This genetic element occurs within the prp8 gene, and is transcribed and translated simultaneously with the gene. After translation, the intein excises itself from the Prp8 protein by an autocatalytic splicing reaction, subsequen...

  6. Bacteriophage-based genetic system for selection of nonsplicing inteins.

    Science.gov (United States)

    Cann, Isaac K O; Amaya, Kensey R; Southworth, Maurice W; Perler, Francine B

    2004-05-01

    A genetic selection system that detects splicing and nonsplicing activities of inteins was developed based on the ability to rescue a T4 phage strain with a conditionally inactive DNA polymerase. This phage defect can be complemented by expression of plasmid-encoded phage RB69 DNA polymerase. Insertion of an intein gene into the active site of the RB69 DNA polymerase gene renders polymerase activity and phage viability dependent on protein splicing. The effectiveness of the system was tested by screening for thermosensitive splicing mutants. Development of genetic systems with the potential of identifying protein splicing inhibitors is a first step towards controlling proliferation of pathogenic microbes harboring inteins in essential proteins.

  7. Intein-modified enzymes, their production and industrial applications

    Energy Technology Data Exchange (ETDEWEB)

    Apgar, James; Lessard, Philip; Raab, Michael R.; Shen, Binzhang; Lazar, Gabor; de la Vega, Humberto

    2016-10-11

    A method of predicting an intein insertion site in a protein that will lead to a switching phenotype is provided. The method includes identifying a plurality of C/T/S sites within the protein; selecting from the plurality of C/T/S/ sites those that are ranked 0.75 or higher by a support vector machine, within ten angstroms of the active site of the protein, and at or near a loop-.beta.-sheet junction or a loop-.alpha.-helix junction. A method of controlling protein activity and hosts including proteins with controlled activity are also provided. Also, intein modified proteins and plants containing intein modified proteins are provided.

  8. Identification of Immunotopes against Mycobacterium leprae as ...

    African Journals Online (AJOL)

    Purpose: To determine the surface epitopes of Mycobacterium leprae (M. leprae) and evaluate their efficacy in the production of anti-M. leprae antibodies in an animal model. Methods: Blood samples were obtained from 34 patients suffering from lepromatous leprosy. Antibodies were obtained from the samples, ...

  9. An intein with genetically selectable markers provides a new approach to internally label proteins with GFP

    Directory of Open Access Journals (Sweden)

    Davis Trisha N

    2011-06-01

    Full Text Available Abstract Background Inteins are proteins that catalyze their own removal from within larger precursor proteins. In the process they splice the flanking protein sequences, termed the N-and C-terminal exteins. Large inteins frequently have a homing endonuclease that is involved in maintaining the intein in the host. Splicing and nuclease activity are independent and distinct domains in the folded structure. We show here that other biochemical activities can be incorporated into an intein in place of the endonuclease without affecting splicing and that these activities can provide genetic selection for the intein. We have coupled such a genetically marked intein with GFP as the N-terminal extein to create a cassette to introduce GFP within the interior of a targeted protein. Results The Pch PRP8 mini-intein of Penicillium chrysogenum was modified to include: 1 aminoglycoside phosphotransferase; 2 imidazoleglycerol-phosphate dehydratase, His5 from S. pombe ; 3 hygromycin B phosphotransferase; and 4 the transcriptional activator LexA-VP16. The proteins were inserted at the site of the lost endonuclease. When expressed in E. coli, all of the modified inteins spliced at high efficiency. Splicing efficiency was also greater than 96% when expressed from a plasmid in S. cerevisiae. In addition the inteins conferred either G418 or hygromycin resistance, or histidine or leucine prototropy, depending on the inserted marker and the yeast genetic background. DNA encoding the marked inteins coupled to GFP as the N-terminal extein was PCR amplified with ends homologous to an internal site in the yeast calmodulin gene CMD1. The DNA was transformed into yeast and integrants obtained by direct selection for the intein's marker. The His5-marked intein yielded a fully functional calmodulin that was tagged with GFP within its central linker. Conclusions Inteins continue to show their flexibility as tools in molecular biology. The Pch PRP8 intein can successfully

  10. Reacciones por lepra en un centro de referencia nacional en Colombia

    Directory of Open Access Journals (Sweden)

    John Nova

    2013-03-01

    Full Text Available Introducción. Colombia es el país de América con mayor proporción de casos nuevos de lepra con discapacidad grave. Para disminuir tal discapacidad se requiere el control de las reacciones, principal causa del daño neural en esta enfermedad. Objetivo. Describir las características clínicas y epidemiológicas y el tratamiento de los pacientes con reacciones de tipo 1 y 2 que consultaron al Centro Dermatológico Federico Lleras Acosta. Materiales y métodos. Se trata de un estudio descriptivo que incluyó la población de pacientes con diagnóstico clínico de reacciones de tipo 1 y de tipo 2 por lepra, que acudieron al centro entre los años 2003 y 2009. Resultados. Se estudiaron 96 reacciones, 35 del tipo 1 y 61 del tipo 2. El 75 % de los pacientes provenía de los departamentos de Tolima, Cundinamarca, Santander y Boyacá. El 56 % de las reacciones de tipo 1 se presentaron antes de iniciar la poliquimioterapia para la lepra; el dermatólogo tratante consideró que las reacciones que se presentaron después de suspender la poliquimioterapia eran recaídas. El 94 % de las reacciones de tipo 1 se trataron con corticoides orales. El 97 % de los pacientes con reacciones de tipo 2 presentaron eritema nudoso, y todos se trataron con talidomida. Conclusiones. La clínica de la reacción de tipo 1 puede orientar al diagnóstico de la lepra en un paciente sin el antecedente de esta enfermedad (56 %. La reacción de tipo 1 que se inicia después de suspender la poliquimioterapia para la lepra, podría ser una manifestación de recaída de la enfermedad. La reacción de tipo 2 es más frecuente en hombres, con una relación hombre a mujer de 4:1. El 97 % de los pacientes con reacción de tipo 2 presentó eritema nudoso.   doi: http://dx.doi.org/10.7705/biomedica.v33i1.582

  11. NMR assignment of a KlbA intein precursor from Methanococcus jannaschii.

    Science.gov (United States)

    Johnson, Margaret A; Southworth, Maurice W; Perler, Francine B; Wüthrich, Kurt

    2007-07-01

    The backbone and side chain resonance assignments of a precursor of the KlbA intein from Methanococcus jannaschii have been determined, based on triple-resonance experiments with the uniformly [13C,15N]-labeled protein.

  12. Mycobacterium leprae virulence-associated peptides are indicators of exposure to M. leprae in Brazil, Ethiopia and Nepal

    DEFF Research Database (Denmark)

    Bobosha, Kidist; Tang, Sheila Tuyet; van der Ploeg-van Schip, Jolien J.

    2012-01-01

    Silent transmission of Mycobacterium leprae, as evidenced by stable leprosy incidence rates in various countries, remains a health challenge despite the implementation of multidrug therapy worldwide. Therefore, the development of tools for the early diagnosis of M. leprae infection should...

  13. Cloning and expression of aequorin photoprotein using intein tag

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    Elah sadat Seyed Hosseini

    2015-01-01

    Full Text Available Background: Intein (INT, is the internal parts of the protein which can be separated from the immature protein during protein splicing process. This sequence requires no specific enzyme or cofactor for separation. This protein sequence and their characteristic of self-cleavage by thiol induction, temperature and pH changes is used for protein purification. The advantage of this method compared to the other protein purification methods is that it doesn’t require any protease enzyme and protease removal steps that make this method important economically. In this study, aequorin photoprotein was hybridized with INT in molecular form and its expression was evaluated. Materials and Methods: In this study, aequorin coding gene that was cloned in pET21-a in the previous studies, was cloned in pTYB21 vector containing INT tag by specific primers and restriction enzymes. Then the resulting pTY-aequarin was transformed to the ER2566 expression strain and cloning accuracy was confirmed by electrophoresis, western blotting and sequencing. Results: The photoprotein aequorin was cloned into SapI/PstI restriction site of pTYB21 plasmid accurately and successfully. Aequorin- INT hybrid protein expression confirmed using traditional methods. Conclusion: The photoprotein aequorin constract in fused with INT confirmed by molecular methods. Also rate of Aequorin- INT expression determined about %25 of cell total protein.

  14. Conservation of intron and intein insertion sites: implications for life histories of parasitic genetic elements

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    Senejani Alireza G

    2009-12-01

    Full Text Available Abstract Background Inteins and introns are genetic elements that are removed from proteins and RNA after translation or transcription, respectively. Previous studies have suggested that these genetic elements are found in conserved parts of the host protein. To our knowledge this type of analysis has not been done for group II introns residing within a gene. Here we provide quantitative statistical support from an analyses of proteins that host inteins, group I introns, group II introns and spliceosomal introns across all three domains of life. Results To determine whether or not inteins, group I, group II, and spliceosomal introns are found preferentially in conserved regions of their respective host protein, conservation profiles were generated and intein and intron positions were mapped to the profiles. Fisher's combined probability test was used to determine the significance of the distribution of insertion sites across the conservation profile for each protein. For a subset of studied proteins, the conservation profile and insertion positions were mapped to protein structures to determine if the insertion sites correlate to regions of functional activity. All inteins and most group I introns were found to be preferentially located within conserved regions; in contrast, a bacterial intein-like protein, group II and spliceosomal introns did not show a preference for conserved sites. Conclusions These findings demonstrate that inteins and group I introns are found preferentially in conserved regions of their respective host proteins. Homing endonucleases are often located within inteins and group I introns and these may facilitate mobility to conserved regions. Insertion at these conserved positions decreases the chance of elimination, and slows deletion of the elements, since removal of the elements has to be precise as not to disrupt the function of the protein. Furthermore, functional constrains on the targeted site make it more difficult

  15. Recombinant production of peptide C-terminal α-amides using an engineered intein

    DEFF Research Database (Denmark)

    Albertsen, Louise; Shaw, Allan C; Norrild, Jens Chr.

    2013-01-01

    of the 198 amino acid intein with an eight amino acid linker. The optimized intein construct was used to produce the PYY derivative under high cell density cultivation conditions, generating the peptide thioester precursor in good yields and subsequent amidation provided the target peptide.......Peptides are of increasing interest as therapeutics in a wide range of diseases, including metabolic diseases such as diabetes and obesity. In the latter, peptide hormones such as peptide YY (PYY) and pancreatic peptide (PP) are important templates for drug design. Characteristic for these peptides...

  16. Exposición Temporal "La Lepra en Colombia"

    OpenAIRE

    Ricardo Rueda González; Hugo Sotomayor

    2001-01-01

    Generalidades sobre la lepra.


    La lepra es una enfermedad infecciosa crónica ocasionada por Mycobateriae /eprae. El principal reservorio y fuente de infección es el enfermo de lepra sin tratamiento. La lepra no es hereditaria, lo que se puede heredar es la susceptibilidad a padecerla.
    Hoy se acepta que la principal vía de transmisión con significación para la salud pública es la vía aére...

  17. pH-dependent activation of Streptomyces hygroscopicus transglutaminase mediated by intein.

    Science.gov (United States)

    Du, Kun; Liu, Zhongmei; Cui, Wenjing; Zhou, Li; Liu, Yi; Du, Guocheng; Chen, Jian; Zhou, Zhemin

    2014-01-01

    Microbial transglutaminase (MTG) from Streptomyces is naturally secreted as a zymogen (pro-MTG), which is then activated by the removal of its N-terminal proregion by additional proteases. Inteins are protein-intervening sequences that catalyze protein splicing without cofactors. In this study, a pH-dependent Synechocystis sp. strain PCC6803 DnaB mini-intein (SDB) was introduced into pro-MTG to simplify its activation process by controlling pH. The recombinant protein (pro-SDB-MTG) was obtained, and the activation process was determined to take 24 h at pH 7 in vitro. To investigate the effect of the first residue in MTG on the activity and the cleavage time, two variants, pro-SDB-MTG(D1S) and pro-SDB-MTG(ΔD1), were expressed, and the activation time was found to be 6 h and 30 h, respectively. The enzymatic property and secondary structure of the recombinant MTG and two variants were similar to those of the wild type, indicating that the insertion of mini-intein did not affect the function of MTG. This insignificant effect was further illustrated by molecular dynamics simulations. This study revealed a controllable and effective strategy to regulate the activation process of pro-MTG mediated by a mini-intein, and it may have great potential for industrial MTG production.

  18. Species-specific detection of Candida tropicalis using evolutionary conserved intein DNA sequences.

    Science.gov (United States)

    Rajasekharan, S K; Ray, A K; Ramesh, S; Kannappan Mohanvel, S

    2018-02-10

    Inteins (internal proteins) are self-splicing transportable genetic elements present in conserved regions of housekeeping genes. The study highlights the importance of intein as a potential diagnostic marker for species-specific identification of Candida tropicalis, a rapidly emerging opportunistic human pathogen. Initial steps of primer validation, sequence alignment, phylogenetic tree analysis, gel electrophoresis and real-time polymerase chain reaction (PCR) assays were performed to confirm the specificity of the designed primers. The primers were selective for C. tropicalis with 100% inclusivity and showed no cross-species or cross-genera matches. The established technique is a prototype for developing multifaceted PCR assays and for point-of-care testing in near future. Development of molecular markers for specific detection of microbial pathogens using real-time polymerase chain reaction (PCR) is an appealing and challenging technique. A real-time PCR is an emerging technology frequently used to detect the aetiologic agents. In recent times, designing species-specific primers for pathogen detection is gaining momentum. The method offers rapid, accurate and cost-effective strategy to identify the target, thus providing sufficient time to instigate appropriate chemotherapy. The study highlights the use of intein DNA sequence as molecular markers for species-specific identification of Candida tropicalis. The study also offers a prototype model for developing multifaceted PCR assays using intein DNA sequences, and provides a developmental starting point for point-of-care testing in near future. © 2018 The Society for Applied Microbiology.

  19. Protein trans-splicing on an M13 bacteriophage: towards directed evolution of a semisynthetic split intein by phage display.

    Science.gov (United States)

    Garbe, Daniel; Thiel, Ilka V; Mootz, Henning D

    2010-10-01

    Split inteins link their fused peptide or protein sequences with a peptide bond in an autocatalytic reaction called protein trans-splicing. This reaction is becoming increasingly important for a variety of applications in protein semisynthesis, polypeptide circularisation, construction of biosensors, or segmental isotopic labelling of proteins. However, split inteins exhibit greatly varying solubility, efficiency and tolerance towards the nature of the fused sequences as well as reaction conditions. We envisioned that phage display as an in vitro selection technique would provide a powerful tool for the directed evolution of split inteins with improved properties. As a first step towards this goal, we show that presentation of active split inteins on an M13 bacteriophage is feasible. Two different C-terminal intein fragments of the Ssp DnaB intein, artificially split at amino acid positions 104 and 11, were encoded in a phagemid vector in fusion to a truncated gpIII protein. For efficient production of hybrid phages, the presence of a soluble domain tag at their N-termini was necessary. Immunoblot analysis revealed that the hybrid phages supported protein trans-splicing with a protein or a synthetic peptide, respectively, containing the complementary intein fragment. Incorporation of biotin or desthiobiotin by this reaction provides a straightforward strategy for future enrichment of desired mutants from randomised libraries of the C-terminal intein fragments on streptavidin beads. Protein semisynthesis on a phage could also be exploited for the selection of chemically modified proteins with unique properties. © 2010 European Peptide Society and John Wiley & Sons, Ltd.

  20. Polymerase chain reaction for the detection of Mycobacterium leprae

    NARCIS (Netherlands)

    Hartskeerl, R. A.; de Wit, M. Y.; Klatser, P. R.

    1989-01-01

    A polymerase chain reaction (PCR) using heat-stable Taq polymerase is described for the specific detection of Mycobacterium leprae, the causative agent of leprosy. A set of primers was selected on the basis of the nucleotide sequence of a gene encoding the 36 kDa antigen of M. leprae. With this set

  1. Mycobacterium leprae virulence-associated peptides are indicators of exposure to M. leprae in Brazil, Ethiopia and Nepal

    DEFF Research Database (Denmark)

    Bobosha, Kidist; Tang, Sheila Tuyet; van der Ploeg-van Schip, Jolien J.

    2012-01-01

    Silent transmission of Mycobacterium leprae, as evidenced by stable leprosy incidence rates in various countries, remains a health challenge despite the implementation of multidrug therapy worldwide. Therefore, the development of tools for the early diagnosis of M. leprae infection should be emph...

  2. New recA mutations that dissociate the various RecA protein activities in Escherichia coli provide evidence for an additional role for RecA protein in UV mutagenesis

    Energy Technology Data Exchange (ETDEWEB)

    Dutreix, M.; Moreau, P.L.; Bailone, A.; Galibert, F.; Battista, J.R.; Walker, G.C.; Devoret, R.

    1989-05-01

    To isolate strains with new recA mutations that differentially affect RecA protein functions, we mutagenized in vitro the recA gene carried by plasmid mini-F and then introduced the mini-F-recA plasmid into a delta recA host that was lysogenic for prophage phi 80 and carried a lac duplication. By scoring prophage induction and recombination of the lac duplication, we isolated new recA mutations. A strain carrying mutation recA1734 (Arg-243 changed to Leu) was found to be deficient in phi 80 induction but proficient in recombination. The mutation rendered the host not mutable by UV, even in a lexA(Def) background. Yet, the recA1734 host became mutable upon introduction of a plasmid encoding UmuD*, the active carboxyl-terminal fragment of UmuD. Although the recA1734 mutation permits cleavage of lambda and LexA repressors, it renders the host deficient in the cleavage of phi 80 repressor and UmuD protein. Another strain carrying mutation recA1730 (Ser-117 changed to Phe) was found to be proficient in phi 80 induction but deficient in recombination. The recombination defect conferred by the mutation was partly alleviated in a cell devoid of LexA repressor, suggesting that, when amplified, RecA1730 protein is active in recombination. Since LexA protein was poorly cleaved in the recA1730 strain while phage lambda was induced, we conclude that RecA1730 protein cannot specifically mediate LexA protein cleavage. Our results show that the recA1734 and recA1730 mutations differentially affect cleavage of various substrates. The recA1730 mutation prevented UV mutagenesis, even upon introduction into the host of a plasmid encoding UmuD* and was dominant over recA+.

  3. Search for Mycobacterium leprae in wild mammals

    Directory of Open Access Journals (Sweden)

    Sílvia Cristina Barboza Pedrini

    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.

  4. Cloning and sequencing the recA+ genes of Acetobacter polyoxogenes and Acetobacter aceti: construction of recA- mutants of by transformation-mediated gene replacement.

    Science.gov (United States)

    Tayama, K; Fukaya, M; Takemura, H; Okumura, H; Kawamura, Y; Horinouchi, S; Beppu, T

    1993-05-15

    The recA+ gene of Acetobacter polyoxogenes was cloned as a gene that conferred methyl methanesulfonate resistance (MMSR) on the RecA- Escherichia coli HB101. The cloned recA+ gene also conferred (i) resistance to UV irradiation, (ii) enhanced intrachromosomal recombination, and (iii) permitted prophage phi 80 induction in E. coli recA- lysogens. Nucleotide sequence determination revealed that the recA product consists of 348 amino acids (aa) corresponding to 38 kDa, and shows significant similarity to RecA proteins from other Gram- bacteria. Next, a portion of recA from Acetobacter aceti was cloned by using polymerase chain reaction with oligodeoxyribonucleotide primers design based on the A. polyoxogenes recA sequence. Due to availability of efficient host-vector and transformation systems in A. aceti, recA mutants of A. aceti were obtained by transformation-mediated gene replacement with the cloned A. aceti recA gene which was inactivated by insertion of the kanamycin-resistance-encoding gene from pACYC177. The recA mutants obtained in this way showed similar phenotypes to those of E. coli recA strains, such as increased sensitivity to MMS and to UV irradiation, and decreased homologous recombination.

  5. The lack of therapeutic effects in mice of the combined gamma-irradiated Mycobacterium leprae and viable BCG against Mycobacterium leprae infection

    International Nuclear Information System (INIS)

    Saito, Hajime; Tomioka, Haruaki; Kitagawa, Toshiyuki

    1985-01-01

    Gamma-irradiated M. leprae in combination with BCG given once biweekly to mice from 2 weeks for up to 187 days after infection with M. leprae caused no significant growth inhibition of M. leprae, at the site of the infection. (author)

  6. Molecular cloning of the recA analog from the marine fish pathogen Vibrio anguillarum 775

    International Nuclear Information System (INIS)

    Singer, J.T.

    1989-01-01

    The recA analog from Vibrio anguillarum 775 was isolated by complementation of recA mutations in Escherichia coli, and its protein product was identified. The recA analog promoted recombination between two partially deleted lactose operons, stimulated both spontaneous and mitomycin C-induced phage production in RecA- lambda lysogens, and restored near wild-type levels of resistance to UV radiation and methyl methanesulfonate

  7. Protein C-Terminal Labeling and Biotinylation Using Synthetic Peptide and Split-Intein

    Science.gov (United States)

    Volkmann, Gerrit; Liu, Xiang-Qin

    2009-01-01

    Background Site-specific protein labeling or modification can facilitate the characterization of proteins with respect to their structure, folding, and interaction with other proteins. However, current methods of site-specific protein labeling are few and with limitations, therefore new methods are needed to satisfy the increasing need and sophistications of protein labeling. Methodology A method of protein C-terminal labeling was developed using a non-canonical split-intein, through an intein-catalyzed trans-splicing reaction between a protein and a small synthetic peptide carrying the desired labeling groups. As demonstrations of this method, three different proteins were efficiently labeled at their C-termini with two different labels (fluorescein and biotin) either in solution or on a solid surface, and a transferrin receptor protein was labeled on the membrane surface of live mammalian cells. Protein biotinylation and immobilization on a streptavidin-coated surface were also achieved in a cell lysate without prior purification of the target protein. Conclusions We have produced a method of site-specific labeling or modification at the C-termini of recombinant proteins. This method compares favorably with previous protein labeling methods and has several unique advantages. It is expected to have many potential applications in protein engineering and research, which include fluorescent labeling for monitoring protein folding, location, and trafficking in cells, and biotinylation for protein immobilization on streptavidin-coated surfaces including protein microchips. The types of chemical labeling may be limited only by the ability of chemical synthesis to produce the small C-intein peptide containing the desired chemical groups. PMID:20027230

  8. Structural studies on Mycobacterium tuberculosis RecA

    Indian Academy of Sciences (India)

    Structures of crystals of Mycobacterium tuberculosis RecA, grown and analysed under different conditions, provide insights into hitherto underappreciated details of molecular structure and plasticity. In particular, they yield information on the invariant and variable features of the geometry of the P-loop, whose binding to ATP ...

  9. Structural studies on Mycobacterium tuberculosis RecA: Molecular ...

    Indian Academy of Sciences (India)

    2015-01-11

    Jan 11, 2015 ... with the protein, characterized in the structures reported here, could be useful for design of inhibitors against. M. tuberculosis RecA. .... rotating anode generator, both using a MAR345 image plate. The crystal to detector ...... structures of Mycobacterium tuberculosis peptidyl-tRNA hydro- lase. J. Mol. Biol.

  10. Structural studies on Mycobacterium tuberculosis RecA: Molecular ...

    Indian Academy of Sciences (India)

    Structures of crystals of Mycobacterium tuberculosis RecA, grown and analysed under different conditions, provide insights into hitherto underappreciated details of molecular structure and plasticity. In particular, they yield information on the invariant and variable features of the geometry of the P-loop, whose binding to ATP ...

  11. La Palabra de Lucio Reca: El Sector Agropecuario

    Directory of Open Access Journals (Sweden)

    Marcelo Coser

    2007-12-01

    Full Text Available Marcelo Coser entrevista al Profesor Lucio Reca en el marco del programa“Prisma Económico” del Instituto de Economía y Finanzas (IEF de la UNC (Entrevista desarrollada el 30 de abril de 2008

  12. Structural studies on Mycobacterium tuberculosis RecA: Molecular ...

    Indian Academy of Sciences (India)

    2015-01-11

    Jan 11, 2015 ... recombination and in the repair of double-strand breaks (Bell. 2005; Cox ... stranded DNA. Its action is triggered by the hydrolysis of bound ATP (McGrew and Knight 2003). RecA is assembled into helical filaments in vitro and into bundles in vivo .... 12 h in the case of suramin, bis-ANS, congo red and bor-.

  13. Dihydropteroate Synthase of Mycobacterium leprae and Dapsone Resistance

    Science.gov (United States)

    Williams, Diana L.; Spring, Laynette; Harris, Eugene; Roche, Paul; Gillis, Thomas P.

    2000-01-01

    Two Mycobacterium leprae genes, folP1 and folP2, encoding putative dihydropteroate synthases (DHPS), were studied for enzymatic activity and for the presence of mutations associated with dapsone resistance. Each gene was cloned and expressed in a folP knockout mutant of Escherichia coli (C600ΔfolP::Kmr). Expression of M. leprae folP1 in C600ΔfolP::Kmr conferred growth on a folate-deficient medium, and bacterial lysates exhibited DHPS activity. This recombinant displayed a 256-fold-greater sensitivity to dapsone (measured by the MIC) than wild-type E. coli C600, and 50-fold less dapsone was required to block (expressed as the 50% inhibitory concentration [IC50]) the DHPS activity of this recombinant. When the folP1 genes of several dapsone-resistant M. leprae clinical isolates were sequenced, two missense mutations were identified. One mutation occurred at codon 53, substituting an isoleucine for a threonine residue (T53I) in the DHPS-1, and a second mutation occurred in codon 55, substituting an arginine for a proline residue (P55R). Transformation of the C600ΔfolP::Kmr knockout with plasmids carrying either the T53I or the P55R mutant allele did not substantially alter the DHPS activity compared to levels produced by recombinants containing wild-type M. leprae folP1. However, both mutations increased dapsone resistance, with P55R having the greatest affect on dapsone resistance by increasing the MIC 64-fold and the IC50 68-fold. These results prove that the folP1 of M. leprae encodes a functional DHPS and that mutations within this gene are associated with the development of dapsone resistance in clinical isolates of M. leprae. Transformants created with M. leprae folP2 did not confer growth on the C600ΔfolP::Kmr knockout strain, and DNA sequences of folP2 from dapsone-susceptible and -resistant M. leprae strains were identical, indicating that this gene does not encode a functional DHPS and is not involved in dapsone resistance in M. leprae. PMID:10817704

  14. An optimized intein-mediated protein ligation approach for the efficient cyclization of cysteine-rich proteins.

    Science.gov (United States)

    Tarasava, Katsiaryna; Freisinger, Eva

    2014-12-01

    Head-to-tail backbone cyclization of proteins is a widely used approach for the improvement of protein stability. One way to obtain cyclic proteins via recombinant expression makes use of engineered Intein tags, which are self-cleaving protein domains. In this approach, pH-induced self-cleavage of the N-terminal Intein tag generates an N-terminal cysteine residue at the target protein, which then attacks in an intramolecular reaction the C-terminal thioester formed by the second C-terminal Intein tag resulting in the release of the cyclic target protein. In the current work we aimed to produce a cyclic analog of the small γ-Ec-1 domain of the wheat metallothionein, which contains six cysteine residues. During the purification process we faced several challenges, among them premature cleavage of one or the other Intein tag resulting in decreasing yields and contamination with linear species. To improve efficiency of the system we applied a number of optimizations such as the introduction of a Tobacco etch virus cleavage site and an additional poly-histidine tag. Our efforts resulted in the production of a cyclic protein in moderate yields without any contamination with linear protein species. © The Author 2014. Published by Oxford University Press. All rights reserved. For Permissions, please e-mail: journals.permissions@oup.com.

  15. High-throughput purification of recombinant proteins using self-cleaving intein tags.

    Science.gov (United States)

    Coolbaugh, M J; Shakalli Tang, M J; Wood, D W

    2017-01-01

    High throughput methods for recombinant protein production using E. coli typically involve the use of affinity tags for simple purification of the protein of interest. One drawback of these techniques is the occasional need for tag removal before study, which can be hard to predict. In this work, we demonstrate two high throughput purification methods for untagged protein targets based on simple and cost-effective self-cleaving intein tags. Two model proteins, E. coli beta-galactosidase (βGal) and superfolder green fluorescent protein (sfGFP), were purified using self-cleaving versions of the conventional chitin-binding domain (CBD) affinity tag and the nonchromatographic elastin-like-polypeptide (ELP) precipitation tag in a 96-well filter plate format. Initial tests with shake flask cultures confirmed that the intein purification scheme could be scaled down, with >90% pure product generated in a single step using both methods. The scheme was then validated in a high throughput expression platform using 24-well plate cultures followed by purification in 96-well plates. For both tags and with both target proteins, the purified product was consistently obtained in a single-step, with low well-to-well and plate-to-plate variability. This simple method thus allows the reproducible production of highly pure untagged recombinant proteins in a convenient microtiter plate format. Copyright © 2016 Elsevier Inc. All rights reserved.

  16. Neuro-lepra: valor de la electromiografia

    Directory of Open Access Journals (Sweden)

    Ernesto Herskovits

    1971-09-01

    Full Text Available Dada la frecuencia con que la lepra afecta al sistema nervioso, consideramos de interés realizar un estudio electromiográfico en zonas corporales clínicamente sanas. Hemos elegido para tal fin 14 enfermos que no tenían lesión sensitivo-motora clínicamente perceptible en el nervio cubital izquierdo. Hemos estudiado tambén un grupo de control de 5 enfermos con lesión evidente del mismo nervio. Se ha comprobado que de los 14 enfermos que aparentemente no tenían lesión del nervio cubital izquierdo, en 12 de ellos surgieron alteraciones electromiográficas que señalan la lesión del nervio, aunque en um grado menor que en el grupo de control. Este hecho nos hace pensar que la agresión que sufre el sistema nervioso periférico es de una extensión mayor que lo hace suponer la clínica, o que las lesiones anatómicas no retrogradan como nos lo sugiere el examen de los pacientes.

  17. Genome-wide comparison of medieval and modern Mycobacterium leprae

    DEFF Research Database (Denmark)

    Schuenemann, Verena J; Singh, Pushpendra; Mendum, Thomas A

    2013-01-01

    Leprosy was endemic in Europe until the Middle Ages. Using DNA array capture, we have obtained genome sequences of Mycobacterium leprae from skeletons of five medieval leprosy cases from the United Kingdom, Sweden, and Denmark. In one case, the DNA was so well preserved that full de novo assembly...

  18. The Leprosy Agents Mycobacterium lepromatosis and Mycobacterium leprae in Mexico

    Science.gov (United States)

    Han, Xiang Y.; Sizer, Kurt Clement; Velarde-Félix, Jesús S.; Frias-Castro, Luis O.; Vargas-Ocampo, Francisco

    2011-01-01

    Summary Background Mycobacterium leprae was the only known cause of leprosy until 2008, when a new species, named Mycobacterium lepromatosis, was found to cause diffuse lepromatous leprosy (DLL), a unique form of leprosy endemic in Mexico. Methods We sought to differentiate the leprosy agents among 120 Mexican patients with various clinical forms of leprosy and to compare their relative prevalence and disease features. Archived skin biopsy specimens from these patients were tested for both M. leprae and M. lepromatosis using polymerase chain reaction-based species-specific assays. Results Eighty-seven (72.5%) patients were confirmed for etiologic species, including 55 with M. lepromatosis, 18 with M. leprae, and 14 with both organisms. The endemic regions of each agent differed but overlapped. Patients with M. lepromatosis were younger and from more states, and their clinical diagnoses included 13 DLL, 34 lepromatous leprosy (LL), and eight other forms of leprosy. By contrast, the diagnoses of patients with M. leprae included none DLL, 15 LL and three other forms. Thus, M. lepromatosis caused DLL specifically (p=0.023). Patients with M. lepromatosis also showed more variable skin lesions and the extremities were the commonest biopsy sites. Finally, patients with dual infections manifested all clinical forms and accounted for 16.1% of all species-confirmed cases. Conclusions M. lepromatosis is another cause of leprosy and is probably more prevalent than M. leprae in Mexico. It mainly causes LL and also specifically DLL. Dual infections caused by both species may occur in endemic area. PMID:22788812

  19. Genetic diversity of mycobacterium leprae isolates from Brazilian leprosy patients.

    Science.gov (United States)

    Fontes, Amanda Nogueira Brum; Sakamuri, Rama Murthy; Baptista, Ida Maria Foschiani Dias; Ura, Somei; Moraes, Milton Ozório; Martínez, Alejandra Nóbrega; Sarno, Euzenir Nunes; Brennan, Patrick J; Vissa, Varalakshmi D; Suffys, Philip Noel

    2009-09-01

    Leprosy is a chronic disease caused by infection with Mycobacterium leprae, an obligate intracellular parasite. A problem in studying the transmission of leprosy is the small amount of variation in bacterial genomic DNA. The discovery of variable number of tandem repeats (VNTRs) allowed the detection of strain variation in areas with a high prevalence of leprosy. Four genotypes of M. leprae based on three single-nucleotide polymorphism (SNPs) were also discovered to be useful for analysis of the global spread of leprosy. In this present study, we examined the allelic diversity of M. leprae at 16 select VNTR and three SNP loci using 89 clinical isolates obtained from patients mainly from the neighbouring states of São Paulo and Rio de Janeiro Brazil. By use of a PCR-RFLP-based procedure that allows the recognition of SNP types 3 and 4 without the need for the more expensive DNA sequencing steps, characterisation of the main M. leprae genotypes was easy. When applied on the study population, it was found that the SNP type 3 is most frequent in these two states of Brazil, and that VNTRs provided further discrimination of the isolates. Two Short Tandem Repeats (STRs) were monomorphic, with the remaining 14 STRs represented by two to 18 alleles. Epidemiological associations with township or state were not evident in this random collection and require further investigations. In phylogenetic trees, branches formed by all 16 STRs clearly separated SNP type 3 organisms from the other types while the allelic patterns of two minisatellite loci 27-5 and 12-5 were highly correlated with SNP type 3. This strain typing study provide the basis for comparison of M. leprae strain types within Brazil and with those from other countries, and informed selection of genomic markers and methods for future studies.

  20. Inteins, introns, and homing endonucleases: recent revelations about the life cycle of parasitic genetic elements

    Directory of Open Access Journals (Sweden)

    Hilario Elena

    2006-11-01

    Full Text Available Abstract Self splicing introns and inteins that rely on a homing endonuclease for propagation are parasitic genetic elements. Their life-cycle and evolutionary fate has been described through the homing cycle. According to this model the homing endonuclease is selected for function only during the spreading phase of the parasite. This phase ends when the parasitic element is fixed in the population. Upon fixation the homing endonuclease is no longer under selection, and its activity is lost through random processes. Recent analyses of these parasitic elements with functional homing endonucleases suggest that this model in its most simple form is not always applicable. Apparently, functioning homing endonuclease can persist over long evolutionary times in populations and species that are thought to be asexual or nearly asexual. Here we review these recent findings and discuss their implications. Reasons for the long-term persistence of a functional homing endonuclease include: More recombination (sexual and as a result of gene transfer than previously assumed for these organisms; complex population structures that prevent the element from being fixed; a balance between active spreading of the homing endonuclease and a decrease in fitness caused by the parasite in the host organism; or a function of the homing endonuclease that increases the fitness of the host organism and results in purifying selection for the homing endonuclease activity, even after fixation in a local population. In the future, more detailed studies of the population dynamics of the activity and regulation of homing endonucleases are needed to decide between these possibilities, and to determine their relative contributions to the long term survival of parasitic genes within a population. Two outstanding publications on the amoeba Naegleria group I intron (Wikmark et al. BMC Evol Biol 2006, 6:39 and the PRP8 inteins in ascomycetes (Butler et al.BMC Evol Biol 2006, 6:42 provide

  1. Branched intermediate formation is the slowest step in the protein splicing reaction of the Ala1 KlbA intein from Methanococcus jannaschii.

    Science.gov (United States)

    Saleh, Lana; Southworth, Maurice W; Considine, Nancy; O'Neill, Colleen; Benner, Jack; Bollinger, J Martin; Perler, Francine B

    2011-12-13

    We report the first detailed investigation of the kinetics of protein splicing by the Methanococcus jannaschii KlbA (Mja KlbA) intein. This intein has an N-terminal Ala in place of the nucleophilic Cys or Ser residue that normally initiates splicing but nevertheless splices efficiently in vivo [Southworth, M. W., Benner, J., and Perler, F. B. (2000) EMBO J.19, 5019-5026]. To date, the spontaneous nature of the cis splicing reaction has hindered its examination in vitro. For this reason, we constructed an Mja KlbA intein-mini-extein precursor using intein-mediated protein ligation and engineered a disulfide redox switch that permits initiation of the splicing reaction by the addition of a reducing agent such as dithiothreitol (DTT). A fluorescent tag at the C-terminus of the C-extein permits monitoring of the progress of the reaction. Kinetic analysis of the splicing reaction of the wild-type precursor (with no substitutions in known nucleophiles or assisting groups) at various DTT concentrations shows that formation of the branched intermediate from the precursor is reversible (forward rate constant of 1.5 × 10(-3) s(-1) and reverse rate constant of 1.7 × 10(-5) s(-1) at 42 °C), whereas the productive decay of this intermediate to form the ligated exteins is faster and occurs with a rate constant of 2.2 × 10(-3) s(-1). This finding conflicts with reports about standard inteins, for which Asn cyclization has been assigned as the rate-determining step of the splicing reaction. Despite being the slowest step of the reaction, branched intermediate formation in the Mja KlbA intein is efficient in comparison with those of other intein systems. Interestingly, it also appears that this intermediate is protected against thiolysis by DTT, in contrast to other inteins. Evidence is presented in support of a tight coupling between the N-terminal and C-terminal cleavage steps, despite the fact that the C-terminal single-cleavage reaction occurs in variant Mja KlbA inteins in

  2. Molecular cloning and characterization of the recA gene of Pseudomonas aeruginosa PAO

    Energy Technology Data Exchange (ETDEWEB)

    Kokjohn, T.A.; Miller, R.V.

    1985-08-01

    The recA gene of Pseudomonas aeruginosa PAO has been isolated and introduced into Escherichia coli K-12. Resistance to killing by UV irradiation was restored in several RecA-E. coli K-12 hosts by the P. aeruginosa gene, as was resistance to methyl methanesulfonate. Recombination proficiency was also restored, as measured by HfrH-mediated conjugation and by the ability to propagate Fec-phage lambda derivatives. The cloned P. aeruginosa recA gene restored both spontaneous and mitomycin C-stimulated induction of lambda prophage in lysogens of a recA strain of E. coli K-12.

  3. Structural and Functional Studies of H. seropedicae RecA Protein – Insights into the Polymerization of RecA Protein as Nucleoprotein Filament

    Energy Technology Data Exchange (ETDEWEB)

    Leite, Wellington C.; Galvão, Carolina W.; Saab, Sérgio C.; Iulek, Jorge; Etto, Rafael M.; Steffens, Maria B.R.; Chitteni-Pattu, Sindhu; Stanage, Tyler; Keck, James L.; Cox, Michael M. (UW); (UW-MED); (Ponta Grossa)

    2016-07-22

    The bacterial RecA protein plays a role in the complex system of DNA damage repair. Here, we report the functional and structural characterization of the Herbaspirillum seropedicae RecA protein (HsRecA). HsRecA protein is more efficient at displacing SSB protein from ssDNA than Escherichia coli RecA protein. HsRecA also promotes DNA strand exchange more efficiently. The three dimensional structure of HsRecA-ADP/ATP complex has been solved to 1.7 Å resolution. HsRecA protein contains a small N-terminal domain, a central core ATPase domain and a large C-terminal domain, that are similar to homologous bacterial RecA proteins. Comparative structural analysis showed that the N-terminal polymerization motif of archaeal and eukaryotic RecA family proteins are also present in bacterial RecAs. Reconstruction of electrostatic potential from the hexameric structure of HsRecA-ADP/ATP revealed a high positive charge along the inner side, where ssDNA is bound inside the filament. The properties of this surface may explain the greater capacity of HsRecA protein to bind ssDNA, forming a contiguous nucleoprotein filament, displace SSB and promote DNA exchange relative to EcRecA. In conclusion, our functional and structural analyses provide insight into the molecular mechanisms of polymerization of bacterial RecA as a helical nucleoprotein filament.

  4. Exposición Temporal "La Lepra en Colombia"

    Directory of Open Access Journals (Sweden)

    Ricardo Rueda González

    2001-12-01

    Full Text Available

    Generalidades sobre la lepra.


    La lepra es una enfermedad infecciosa crónica ocasionada por Mycobateriae /eprae. El principal reservorio y fuente de infección es el enfermo de lepra sin tratamiento. La lepra no es hereditaria, lo que se puede heredar es la susceptibilidad a padecerla.
    Hoy se acepta que la principal vía de transmisión con significación para la salud pública es la vía aérea, a través del tracto respiratorio superior, aunque se reconocen como eventuales puertas de entrada la piel y el tracto digestivo. Por esto, el principal factor de riesgo para adquirirla es el contacto o convivencia con un paciente bacilífero. El periodo de incubación para la forma multibacilar sería de 8 a 12 años. y para la forma paucibacilar entre 2 y 5 años.
    Se sabe que la forma tuberculoide predomina en personas con piel negra. En Colombia, en las región del Chocó, predominantemente habitados por personas de piel negra, predomina la lepra tuberculoide ( la menos contagiosa por ser paucibacilar. La transmisión por vía aérea, la colonización bacilar sistémica, originaría las formas multibacilares, mientras la entrada del bacilo por la vía cutánea predispondría a las formas paucibacilares.
    Aunque hoy se sabe que los estudios experimentales han demostrado la posibilidad de que algunos artrópodos hematófagos pueden trasmitir experimentalmente la lepra, esta evidencia no es suficiente para saber que la naturaleza utilice esta vía.
    La entrada del Mycobateriae /eprae al organismo en el 90% de las personas lleva a una infección subclínica y a la cura sin tratamiento. Esta resistencia natural  proviene de la respuesta inmune, de la magnitud y de la frecuencia de la exposición al bacilo. La susceptibilidad inmunológica del huésped, la magnitud del inoculó y las necesidad básicas insatisfechas, desnutrición, hacinamiento, mala higiene personal y de la

  5. Cost effective purification of intein based syntetic cationic antimicrobial peptide expressed in cold shock expression system using salt inducible E. coli GJ1158

    Directory of Open Access Journals (Sweden)

    Seetha Ram Kotra

    2014-03-01

    Full Text Available Objective:Synthetic cationic antimicrobial peptide (SC-AMP is an important and upcoming therapeutic molecule against onventional antibiotics. In this study, an attempt was made to purify the SC-AMP without the enzymatic cleavage of the affinity tag, by using an intein-based system. Methods:The intein sequence was amplified from pTYB11 vector using PCR methodologies and the N-terminal of intein was ligated with SC-AMP. The designed construct, intein-SC-AMP was cloned into MCS region of cold shock expression vector, pCOLDI and the recombinant peptide was purified on a chitin affinity column by cleaving intein with 50 mM DTT without applying enzymatic cleavage. Later the peptide was quantified and its antibacterial activity of the purified peptide was studied using well diffusion method. Results: Initially, intein-SC-AMP was expressed as a fusion protein in both IPTG inducible E. coli BL21(DE3 and salt inducible E. coli GJ1158. Single step purification using CBD (chitin binding domain - intein tag in salt inducible E. coli GJ1158, yields the SC-AMP in the soluble form at a oncentration of 208 mg/L. The antibacterial activity and minimal inhibitory concentration (MIC of the purified SC-AMP was studied against both Gram positive and Gram negative microorganisms. Conclusion: For the first time, single step purification of soluble SC-AMP was carried out using chitin-binding domain affinity tag in salt inducible E. coli GJ1158 without an application of enzymatic cleavage. J Microbiol Infect Dis 2014;4(1:13-19

  6. Rare Atypical Presentations in Type 2 Lepra Reaction: A Case Series

    OpenAIRE

    Mitra, Debdeep

    2017-01-01

    Abstract Background Leprosy or Hansen’s disease is a chronic mycobacterial infection caused by mycobacterium leprae. Reactions in Leprosy represent an acute inflammataory response during the natural chronic course of infection. Type 2 lepra reaction is a Th2-mediated type III hypersensitivity reaction in leprosy, with a characteristic cutaneous manifestation in the form of erythema nodosum leprosum (ENL). This presentation describes unusual presentations of Type 2 lepra reaction in five patie...

  7. ADAKAH KAITAN ANTARA PENYAKIT KUSTA KERBAU (Lepra bubalorum DAN PENYAKIT KUSTA MANUSIA (Lepra humanum DI SULAWESI ?

    Directory of Open Access Journals (Sweden)

    Iwan T. Budiarso

    2012-09-01

    Full Text Available Lepra bubalorum or leprosy in water buffaloes, is an exotic disease in buffaloes. It is a very interesting phenomenon in Veterinary Medicine because until to date it has been found only in Indonesia. Kok and Rusli (1926 has published their first report in 1926 describing the disease in Java. Since then the diagnosis of leprosy has been established by histological examinations in about 146 water buffaloes (Lobel, 1934. It has also been found in Holstein-Freisen cow (Ressang and Titus, I960, one case in Ongole breed and another in an unidentified cow breed. For leprosy infection in bovine, Kraneveld and Roza (1954 proposed to use the name Lepra bovina Lobel (1934 was the first scientist who brought the attention of this unusual disease to the international scientific world. His extensive works gave vs a clear description of the clinical signs and symptoms, histopathological changes and the microbiological arrangement of the acid fast baccili of the infectious agent in the affected tissues. The changes found in the cutaneous tissue of these animals resembled to those found in the skin of lepromatous lessions in humans. The rod-shape bacteria which is acid fast, are always present in the nodules of the diseases animals. In central Sulawesi (Celebes, leprosy in humans was found as the second most prevalant chronic diseases after pulmonary tuberculosis. It is interesting to know that apparently human leprosy was also found in great numbers in the same area where leprosy in water buffaloes was found frequently. This phenomenon is an interesting subject for investigation to see whether the genus of the bacteria found in humans and that in water buffaloes have the anthropozoonotic properties. *    Makalah ini disajikan pada Simposium "Kemajuan Dalam Penyakit Tropis dan Parasit" di Fakultas Kedokteran, Universitas Tarumanagara, Jakarta, 12 Desember 1992 **   Bagian Patologi Anatomi, Fakultas Kedokteran Universitas Tarumanagara, Jakarta, Indonesia. Up

  8. A study on PCR for detecting infection with M. leprae.

    Science.gov (United States)

    Qinxue, W; Xinyu, L; Wei, H; Tao, L; Yaoping, Y; Jinping, Z; Xiuling, C; Ganyun, Y

    1999-12-01

    So far, it has not been established a satisfactory method for early diagnosis and studying on epidemiology for leprosy, we want to develop a molecular biological method for solving this point. Based on the M. leprae gene coding groEL, 65 kD and 16S rRNA, three polymerase chain reactions were developed by using Plikaytis', Woods' and Pattyn's procedures. It was optimized that the experimental parameters for each PCR, and a comparative study on practivity among three PCRs was also conducted for practical purpose. For detecting infection with M. leprae, all of PCRs established by us were highly sensitive and specific, but for practical purpose, the Woods' PCR optimized by us ought to be chosen firstly.

  9. Characterization of the Pseudomonas aeruginosa recA analog and its protein product: rec-102 is a mutant allele of the P. aeruginosa PAO recA gene

    Energy Technology Data Exchange (ETDEWEB)

    Kokjohn, T.A.; Miller, R.V.

    1987-04-01

    We cloned a 2.3-kilobase-pair fragment of the Pseudomonas aeruginosa PAO chromosome which is capable of complementing recA mutations of Escherichia coli. The recA-complementing activity was further localized to a 1.5-kilobase-pair PvuII-HindIII fragment. Southern blot analysis under conditions of high stringency indicated that DNA sequence homology is shared by the E. coli recA gene and the P. aeruginosa recA analog. The cloned recA analog was shown to restore resistance to methyl methanesulfonate, nitrofurantoin, and UV irradiation to E. coli recA mutants. Upon introduction of the cloned P. aeruginosa gene, these mutants regained recombination proficiency in HfrH-mediated conjugation and the ability to induce lambda prophages and SOS functions (din gene transcription) after exposure to DNA-damaging agents. Lambda prophage carrying a cI ind mutation was not inducible, suggesting that the mechanism of induction of these SOS functions by the P. aeruginosa RecA analog is similar to that by the activated E. coli RecA protein. The product of the recA analog was identified in minicells as a protein of approximately 47,000 daltons. Western blot analysis using anti-E. coli RecA antibody demonstrated that this protein is antigenically cross-reactive with the E. coli recA protein. The recA-containing fragment was cloned into the broad-host-range vector pCP13 and introduced into Rec- strains of P. aeruginosa containing the rec-102 allele. The plasmid was shown to restore recombination proficiency in FP5-mediated conjugations and to restore resistance to UV irradiation and methyl methanesulfonate to these Rec- mutants. It was shown that a wild-type allele of rec-102 is necessary for UV-mediated induction of D3 and F116 prophages. The cloned recA analog restored the UV inducibility of these prophages in rec-102 mutants.

  10. Dynamics of RecA filaments on single-stranded DNA

    NARCIS (Netherlands)

    Van Loenhout, M.T.J.; Van der Heijden, T.; Kanaar, R.; Wyman, C.; Dekker, C.

    2009-01-01

    RecA, the key protein in homologous recombination, performs its actions as a helical filament on single-stranded DNA (ssDNA). ATP hydrolysis makes the RecA–ssDNA filament dynamic and is essential for successful recombination. RecA has been studied extensively by single-molecule techniques on

  11. Complementation pattern of lexB and recA mutations in Escherichia coli K12; mapping of tif-1, lexB and recA mutations

    International Nuclear Information System (INIS)

    Morand, P.; Goze, A.; Devoret, R.

    1977-01-01

    Three lexB mutations, whose phenotypes have been previously characterized, are studied here in relation to a few recA mutations as to their complementation pattern and relative location. The restoration of resistance to UV-light and to X-rays in the hetero-allelic diploid bacteria was used as a test for dominance and complementation. The wild type allele was always dominant over the mutant allele. Only partial complementation was found between lexB and two rexA alleles. There was no complementation between the recA alleles. All the data taken together strongly suggest that the complementations found are intragenic: lexB and recA mutations are in one gene. Mapping of lexB, recA and tif-1 mutations in relation to srl-1 and cysC by phage P1 transduction shows that lexB and the tif-1 mutations form a cluster proximal to srl-1 whereas recA mutations are located at the other extremity of the gene. Variability with temperature of cotransduction frequencies as well as their extended range of values prevent a meaningful calculation of the length of the recA gene. Our hypothesis is that the recA protein has two functional regions called A and B respectively defined at the genetical level by recA and lexB mutations and that it is, in vivo, an oligomeric protein forming a complex with the lexA protein. This complex is postulated to be multifunctional: recombination and control of exonuclease V are effected by the A region while the B region and lexA protein effect induced DNA repair and lysogenic induction. (orig.) [de

  12. Further biochemical characterization of Mycobacterium leprae laminin-binding proteins

    Directory of Open Access Journals (Sweden)

    M.A.M. Marques

    2001-04-01

    Full Text Available It has been demonstrated that the alpha2 chain of laminin-2 present on the surface of Schwann cells is involved in the process of attachment of Mycobacterium leprae to these cells. Searching for M. leprae laminin-binding molecules, in a previous study we isolated and characterized the cationic proteins histone-like protein (Hlp and ribosomal proteins S4 and S5 as potential adhesins involved in M. leprae-Schwann cell interaction. Hlp was shown to bind alpha2-laminins and to greatly enhance the attachment of mycobacteria to ST88-14 Schwann cells. In the present study, we investigated the laminin-binding capacity of the ribosomal proteins S4 and S5. The genes coding for these proteins were PCR amplified and their recombinant products were shown to bind alpha2-laminins in overlay assays. However, when tested in ELISA-based assays and in adhesion assays with ST88-14 cells, in contrast to Hlp, S4 and S5 failed to bind laminin and act as adhesins. The laminin-binding property and adhesin capacity of two basic host-derived proteins were also tested, and only histones, but not cytochrome c, were able to increase bacterial attachment to ST88-14 cells. Our data suggest that the alanine/lysine-rich sequences shared by Hlp and eukaryotic H1 histones might be involved in the binding of these cationic proteins to laminin.

  13. Using intein catalysis to probe the origin of major histocompatibility complex class I-presented peptides

    Science.gov (United States)

    Farfán-Arribas, Diego J.; Stern, Lawrence J.; Rock, Kenneth L.

    2012-01-01

    All vertebrate nucleated cells generate peptides from their expressed gene products and then display them at the cell surface bound to MHC class I molecules. This allows CD8+ T cells to detect and eliminate abnormal cells that are synthesizing foreign proteins, e.g., from viruses or mutations. To permit the immune system to more uniformly monitor a cell's proteins, regardless of their half-life or location, it has been thought that the products of rapid degradation of the mistakes of protein synthesis (defective ribosomal products, DRiPs) preferentially contribute to the class I-presented peptides. However, using intein catalysis to generate peptide sequences exclusively by posttranslational splicing of mature proteins, we show here that presented peptides can be generated from fully folded and functional proteins. Remarkably, the presentation of peptides from two model mature proteins is just as efficient as from newly synthesized proteins subject to errors in translation or folding. These results indicate that for the constructs we have analyzed, DRiPs are not a more efficient source of class I peptides for antigen presentation than the turnover of mature functional proteins. Accordingly, our data suggest that one of the major ways the immune system evaluates the health of cells is by monitoring the breakdown products of the proteome. PMID:23027972

  14. Intein-mediated Cre protein assembly for transgene excision in hybrid progeny of transgenic Arabidopsis.

    Science.gov (United States)

    Ge, Jia; Wang, Lijun; Yang, Chen; Ran, Lingyu; Wen, Mengling; Fu, Xianan; Fan, Di; Luo, Keming

    2016-10-01

    An approach for restoring recombination activity of complementation split-Cre was developed to excise the transgene in hybrid progeny of GM crops. Growing concerns about the biosafety of genetically modified (GM) crops has currently become a limited factor affecting the public acceptance. Several approaches have been developed to generate selectable-marker-gene-free GM crops. However, no strategy was reported to be broadly applicable to hybrid crops. Previous studies have demonstrated that complementation split-Cre recombinase restored recombination activity in transgenic plants. In this study, we found that split-Cre mediated by split-intein Synechocystis sp. DnaE had high recombination efficiency when Cre recombinase was split at Asp232/Asp233 (866 bp). Furthermore, we constructed two plant expression vectors, pCA-NCre-In and pCA-Ic-CCre, containing NCre866-In and Ic-CCre866 fragments, respectively. After transformation, parent lines of transgenic Arabidopsis with one single copy were generated and used for hybridization. The results of GUS staining demonstrated that the recombination activity of split-Cre could be reassembled in these hybrid progeny of transgenic plants through hybridization and the foreign genes flanked by two loxP sites were efficiently excised. Our strategy may provide an effective approach for generating the next generation of GM hybrid crops without biosafety concerns.

  15. Lepra lepromatosa . A propósito de un caso clínico (Leprae lepromatous. A case report

    Directory of Open Access Journals (Sweden)

    Anays Toro

    2014-08-01

    Full Text Available Resumen (español La lepra es una enfermedad infectocontagiosa, crónica, declarada por la OMS como exitosamente controlada desde hace varios años, cuyo agente causal es el bacilo ácido alcohol resistente (BAAR Mycobacterium leprae, La infección tiene un largo período de incubación y afecta principalmente piel, mucosas y nervios periféricos. Su presentación clínica comprende dos tipos polares, la Lepra lepromatosa (LL, la Lepra tuberculoide (LT y tres expresiones intermedias. En este trabajo, se presenta el caso de un paciente masculino de 36 años, quien desde hace dos años cursa con maculas cutáneas, hipercrómicas, que progresaron a nódulos, inicialmente en manos y pies, luego se extendieron al resto del cuerpo, sin respetar palma de manos y planta de pies. Precedido por una infección por Virus del Dengue (20 días antes. Al momento de ingreso el paciente cursaba con lesiones nodulares, multiformes, confluentes, induradas e incontables; sin alteraciones de la sensibilidad superficial o profunda, ni neuromusculares. Estudio radiológicos de tórax, abdomen y pelvis no evidenciaron lesiones osteomusculares o viscerales, ni compromiso ganglionar. La segunda biopsia reportó inflamación crónica, infiltrado linfocitario escaso, macrófagos espumosos e incontables BAAR, ligeramente curvados, intracelulares (macrófagos. Prueba de lepromina fue negativa. El diagnóstico final fue lepra Lepromatosa. Se prescribió multiterapia triple (Dapsona, Rifampicina, Clofazimina evidenciándose una mejoría en el número y tamaño de las lesiones a los dos meses de iniciada la terapia. El diagnóstico precoz a través de parámetros clínicos, histopatológico, inmunológicos y baciloscópicos son fundamentales en el amplio marco de las presentaciones clínicas en la Lepra. Abstract (english Leprosy is a chronic infectious disease, declared by the WHO as successfully controlled since several years ago, is causal by an acid

  16. Tuberculosis vaccine strain Mycobacterium bovis BCG Russia is a natural recA mutant

    Directory of Open Access Journals (Sweden)

    Böttger Erik C

    2008-07-01

    Full Text Available Abstract Background The current tuberculosis vaccine is a live vaccine derived from Mycobacterium bovis and attenuated by serial in vitro passaging. All vaccine substrains in use stem from one source, strain Bacille Calmette-Guérin. However, they differ in regions of genomic deletions, antigen expression levels, immunogenicity, and protective efficacy. Results As a RecA phenotype increases genetic stability and may contribute restricting the ongoing evolution of the various BCG substrains while maintaining their protective efficacy, we aimed to inactivate recA by allelic replacement in BCG vaccine strains representing different phylogenetic lineages (Pasteur, Frappier, Denmark, Russia. Homologous gene replacement was achieved successfully in three out of four strains. However, only illegitimate recombination was observed in BCG substrain Russia. Sequence analyses of recA revealed that a single nucleotide insertion in the 5' part of recA led to a translational frameshift with an early stop codon making BCG Russia a natural recA mutant. At the protein level BCG Russia failed to express RecA. Conclusion According to phylogenetic analyses BCG Russia is an ancient vaccine strain most closely related to the parental M. bovis. We hypothesize that recA inactivation in BCG Russia occurred early and is in part responsible for its high degree of genomic stability, resulting in a substrain that has less genetic alterations than other vaccine substrains with respect to M. bovis AF2122/97 wild-type.

  17. The RecX protein interacts with the RecA protein and modulates its activity in Herbaspirillum seropedicae

    International Nuclear Information System (INIS)

    Galvão, C.W.; Souza, E.M.; Etto, R.M.; Pedrosa, F.O.; Chubatsu, L.S.; Yates, M.G.; Schumacher, J.; Buck, M.; Steffens, M.B.R.

    2012-01-01

    DNA repair is crucial to the survival of all organisms. The bacterial RecA protein is a central component in the SOS response and in recombinational and SOS DNA repairs. The RecX protein has been characterized as a negative modulator of RecA activity in many bacteria. The recA and recX genes of Herbaspirillum seropedicae constitute a single operon, and evidence suggests that RecX participates in SOS repair. In the present study, we show that the H. seropedicae RecX protein (RecX Hs ) can interact with the H. seropedicae RecA protein (RecA Hs ) and that RecA Hs possesses ATP binding, ATP hydrolyzing and DNA strand exchange activities. RecX Hs inhibited 90% of the RecA Hs DNA strand exchange activity even when present in a 50-fold lower molar concentration than RecA Hs . RecA Hs ATP binding was not affected by the addition of RecX, but the ATPase activity was reduced. When RecX Hs was present before the formation of RecA filaments (RecA-ssDNA), inhibition of ATPase activity was substantially reduced and excess ssDNA also partially suppressed this inhibition. The results suggest that the RecX Hs protein negatively modulates the RecA Hs activities by protein-protein interactions and also by DNA-protein interactions

  18. Autogenous regulation and kinetics of induction of Pseudomonas aeruginosa recA transcription as analyzed with operon fusions

    International Nuclear Information System (INIS)

    Horn, J.M.; Ohman, D.E.

    1988-01-01

    A promoterless chloramphenicol acetyltransferase gene (cat) was used to construct recA-cat operon fusions to quantitatively examine the transcriptional regulation of the Pseudomonas aeruginosa recA gene in P. aeruginosa PAO. Wild-type P. aeruginosa containing the recA8-cat fusion was treated with methyl methanesulfonate (MMS) and showed immediate induction of chloramphenicol acetyltransferase (CAT) specific activity, whereas a recA::Tn501 mutant of P. aeruginosa containing recA8-cat showed no induction with MMS. This indicated that a functional copy of recA was required for derepression of recA transcription and that P. aeruginosa recA protein was a positive regulatory factor promoting its own expression. Compared with that in the wild type, the uninduced level of CAT in recA8-cat-containing cells was reduced by approximately one-half in the recA::Tn501 mutant, indicating that recA+-dependent spontaneous induction contributes to the uninduced levels of recA expression in P. aeruginosa. MMS (0.012%) caused recA-directed CAT synthesis to increase almost immediately, with maximum CAT activity, fourfold higher than uninduced levels, attained at 60 min postinduction. The kinetics of recA8-cat fusion activity were shown to be directly related to the MMS doses used. Another fusion called recAa1-cat, where cat was located between the two transcriptional terminators of the P. aeruginosa recA gene, also showed dose-dependent induction by MMS, but the CAT activity from recAa1-cat was only one-half of that obtained with recA8-cat under the same conditions. Treatment of recA+ P. aeruginosa containing recA8-cat with UV irradiation produced an immediate effect on recA8-cat transcription and showed little UV dose dependency at doses of 5 J/m2 or greater

  19. Sensitization or tolerance to Mycobacterium leprae antigen by route of injection

    International Nuclear Information System (INIS)

    Shepard, C.C.; Walker, L.L.; Van Landingham, R.M.; Ye, S.Z.

    1982-01-01

    Aqueous suspensions of heat-killed Mycobacterium leprae in a dose of 10(7) organisms were highly immunogenic when injected intradermally (i.d.). The same dose of bacteria did not sensitize when given intraperitoneally (i.p.) or intravenously (i.v.), and did so only minimally at best when given subcutaneously. The i.d. route was the most immunogenic for sheep erythrocytes also. M. leprae injected i.p. or i.v. stimulated immune tolerance to M. leprae challenge i.d. In older mice (greater than or equal to 8 weeks), the i.v. injections gave more complete tolerance. Mice that had been rendered tolerant by i.v. injections maintained their tolerance for at least 168 days. Prior UV irradiation of intact mice prevented sensitization by the i.d. route. In normal mice, living M. bovis BCG given i.d. produced good sensitization to M. leprae. Mice that had been made tolerant by i.v. injection of M. leprae could be partially sensitized to M. leprae by i.d. immunization with BCG; mixtures of living BCG and heat-killed M. leprae were no more effective than BCG alone. These findings appear to have relevance to the pathogenesis of lepromatous leprosy and its immunoprophylaxis

  20. RecA: Regulation and Mechanism of a Molecular Search Engine.

    Science.gov (United States)

    Bell, Jason C; Kowalczykowski, Stephen C

    2016-06-01

    Homologous recombination maintains genomic integrity by repairing broken chromosomes. The broken chromosome is partially resected to produce single-stranded DNA (ssDNA) that is used to search for homologous double-stranded DNA (dsDNA). This homology driven 'search and rescue' is catalyzed by a class of DNA strand exchange proteins that are defined in relation to Escherichia coli RecA, which forms a filament on ssDNA. Here, we review the regulation of RecA filament assembly and the mechanism by which RecA quickly and efficiently searches for and identifies a unique homologous sequence among a vast excess of heterologous DNA. Given that RecA is the prototypic DNA strand exchange protein, its behavior affords insight into the actions of eukaryotic RAD51 orthologs and their regulators, BRCA2 and other tumor suppressors. Copyright © 2016 Elsevier Ltd. All rights reserved.

  1. Alternate radiolabeled markers for detecting metabolic activity of Mycobacterium leprae residing in murine macrophages

    International Nuclear Information System (INIS)

    Prasad, H.K.; Hastings, R.C.

    1985-01-01

    This study demonstrated the utility of using 4% NaOH as a murine macrophage cell-solubilizing agent to discriminate between host macrophage metabolism and that of intracellular Mycobacterium leprae. A 4% concentration of NaOH had no deleterious effect on labeled mycobacteria. Thereby, alternate radiolabeled indicators of the metabolic activity of intracellular M. leprae could be experimented with. Significant incorporation of 14 C-amino acid mixture, [ 14 C]leucine, [ 14 C]uridine, and carrier-free 32 P was observed in cultures containing freshly extracted (''live'') strains of M. leprae as compared with control cultures containing autoclaved bacilli

  2. Identification of mimotopes of Mycobacterium leprae as potential diagnostic reagents

    Directory of Open Access Journals (Sweden)

    Alban Silvana M

    2013-01-01

    Full Text Available Abstract Background An early diagnostic test for detecting infection in leprosy is fundamental for reducing patients’ sequelae. The currently used lepromin is not adequate for disease diagnosis and, so far, no antigen to be used in intradermoreaction has proved to be sensitive and specific for that purpose. Aiming at identifying new reagents to be used in skin tests, candidate antigens were investigated. Methods Random peptide phage display libraries were screened by using antibodies from leprosy patients in order to identify peptides as diagnostic reagents. Results Seven different phage clones were identified using purified antibodies pooled from sera of leprosy patients. When the clones were tested with serum samples by ELISA, three of them, 5A, 6A and 1B, allowed detecting a larger number of leprosy patients when compared to controls. The corresponding peptides expressed by selected phage clones were chemically synthesized. A pilot study was undertaken to assess the use of peptides in skin tests. The intradermal challenge with peptides in animals previously sensitized with Mycobacterium leprae induced a delayed-type hypersensitivity with peptide 5A (2/5 and peptide 1B (1/5. In positive controls, there was a 3/5 reactivity for lepromin and a 4/5 reactivity of the sensitized animals with soluble extract of M. leprae. Conclusions The preliminary data suggest that may be possible to develop reagents with diagnostic potential based on peptide mimotopes selected by phage display using polyclonal human antibodies.

  3. Phylogenomics and antimicrobial resistance of the leprosy bacillus Mycobacterium leprae.

    Science.gov (United States)

    Benjak, Andrej; Avanzi, Charlotte; Singh, Pushpendra; Loiseau, Chloé; Girma, Selfu; Busso, Philippe; Fontes, Amanda N Brum; Miyamoto, Yuji; Namisato, Masako; Bobosha, Kidist; Salgado, Claudio G; da Silva, Moisés B; Bouth, Raquel C; Frade, Marco A C; Filho, Fred Bernardes; Barreto, Josafá G; Nery, José A C; Bührer-Sékula, Samira; Lupien, Andréanne; Al-Samie, Abdul R; Al-Qubati, Yasin; Alkubati, Abdul S; Bretzel, Gisela; Vera-Cabrera, Lucio; Sakho, Fatoumata; Johnson, Christian R; Kodio, Mamoudou; Fomba, Abdoulaye; Sow, Samba O; Gado, Moussa; Konaté, Ousmane; Stefani, Mariane M A; Penna, Gerson O; Suffys, Philip N; Sarno, Euzenir Nunes; Moraes, Milton O; Rosa, Patricia S; Baptista, Ida M F Dias; Spencer, John S; Aseffa, Abraham; Matsuoka, Masanori; Kai, Masanori; Cole, Stewart T

    2018-01-24

    Leprosy is a chronic human disease caused by the yet-uncultured pathogen Mycobacterium leprae. Although readily curable with multidrug therapy (MDT), over 200,000 new cases are still reported annually. Here, we obtain M. leprae genome sequences from DNA extracted directly from patients' skin biopsies using a customized protocol. Comparative and phylogenetic analysis of 154 genomes from 25 countries provides insight into evolution and antimicrobial resistance, uncovering lineages and phylogeographic trends, with the most ancestral strains linked to the Far East. In addition to known MDT-resistance mutations, we detect other mutations associated with antibiotic resistance, and retrace a potential stepwise emergence of extensive drug resistance in the pre-MDT era. Some of the previously undescribed mutations occur in genes that are apparently subject to positive selection, and two of these (ribD, fadD9) are restricted to drug-resistant strains. Finally, nonsense mutations in the nth excision repair gene are associated with greater sequence diversity and drug resistance.

  4. The RecX protein interacts with the RecA protein and modulates its activity in Herbaspirillum seropedicae

    Directory of Open Access Journals (Sweden)

    C.W. Galvão

    2012-12-01

    Full Text Available DNA repair is crucial to the survival of all organisms. The bacterial RecA protein is a central component in the SOS response and in recombinational and SOS DNA repairs. The RecX protein has been characterized as a negative modulator of RecA activity in many bacteria. The recA and recX genes of Herbaspirillum seropedicae constitute a single operon, and evidence suggests that RecX participates in SOS repair. In the present study, we show that the H. seropedicae RecX protein (RecX Hs can interact with the H. seropedicaeRecA protein (RecA Hs and that RecA Hs possesses ATP binding, ATP hydrolyzing and DNA strand exchange activities. RecX Hs inhibited 90% of the RecA Hs DNA strand exchange activity even when present in a 50-fold lower molar concentration than RecA Hs. RecA Hs ATP binding was not affected by the addition of RecX, but the ATPase activity was reduced. When RecX Hs was present before the formation of RecA filaments (RecA-ssDNA, inhibition of ATPase activity was substantially reduced and excess ssDNA also partially suppressed this inhibition. The results suggest that the RecX Hs protein negatively modulates the RecA Hs activities by protein-protein interactions and also by DNA-protein interactions.

  5. Characterization of a Mycobacterium leprae antigen related to the secreted Mycobacterium tuberculosis protein MPT32

    NARCIS (Netherlands)

    Wieles, B.; van Agterveld, M.; Janson, A.; Clark-Curtiss, J.; Rinke de Wit, T.; Harboe, M.; Thole, J.

    1994-01-01

    Secreted proteins may serve as major targets in the immune response to mycobacteria. To identify potentially secreted Mycobacterium leprae antigens, antisera specific for culture filtrate proteins of Mycobacterium tuberculosis were used to screen a panel of recombinant antigens selected previously

  6. Experimental Infection of Rhodnius prolixus (Hemiptera, Triatominae with Mycobacterium leprae Indicates Potential for Leprosy Transmission.

    Directory of Open Access Journals (Sweden)

    Arthur da Silva Neumann

    Full Text Available Leprosy is a chronic dermato-neurological disease caused by infection with Mycobacterium leprae. In 2013 almost 200,000 new cases of leprosy were detected around the world. Since the first symptoms take from years to decades to appear, the total number of asymptomatic patients is impossible to predict. Although leprosy is one of the oldest records of human disease, the mechanisms involved with its transmission and epidemiology are still not completely understood. In the present work, we experimentally investigated the hypothesis that the mosquitoes Aedes aegypti and Culex quinquefasciatus and the hemiptera Rhodnius prolixus act as leprosy vectors. By means of real-time PCR quantification of M. leprae 16SrRNA, we found that M. leprae remained viable inside the digestive tract of Rhodnius prolixus for 20 days after oral infection. In contrast, in the gut of both mosquito species tested, we were not able to detect M. leprae RNA after a similar period of time. Inside the kissing bug Rhodnius prolixus digestive tract, M. leprae was initially restricted to the anterior midgut, but gradually moved towards the hindgut, in a time course reminiscent of the life cycle of Trypanosoma cruzi, a well-known pathogen transmitted by this insect. The maintenance of M. leprae infectivity inside the digestive tract of this kissing bug is further supported by successful mice footpad inoculation with feces collected 20 days after infection. We conclude that Rhodnius prolixus defecate infective M. leprae, justifying the evaluation of the presence of M. leprae among sylvatic and domestic kissing bugs in countries endemic for leprosy.

  7. Long-term survival and virulence of Mycobacterium leprae in amoebal cysts.

    Directory of Open Access Journals (Sweden)

    William H Wheat

    2014-12-01

    Full Text Available Leprosy is a curable neglected disease of humans caused by Mycobacterium leprae that affects the skin and peripheral nerves and manifests clinically in various forms ranging from self-resolving, tuberculoid leprosy to lepromatous leprosy having significant pathology with ensuing disfiguration disability and social stigma. Despite the global success of multi-drug therapy (MDT, incidences of clinical leprosy have been observed in individuals with no apparent exposure to other cases, suggestive of possible non-human sources of the bacteria. In this study we show that common free-living amoebae (FLA can phagocytose M. leprae, and allow the bacillus to remain viable for up to 8 months within amoebic cysts. Viable bacilli were extracted from separate encysted cocultures comprising three common Acanthamoeba spp.: A. lenticulata, A. castellanii, and A. polyphaga and two strains of Hartmannella vermiformis. Trophozoites of these common FLA take up M. leprae by phagocytosis. M. leprae from infected trophozoites induced to encyst for long-term storage of the bacilli emerged viable by assessment of membrane integrity. The majority (80% of mice that were injected with bacilli extracted from 35 day cocultures of encysted/excysted A. castellanii and A. polyphaga showed lesion development that was similar to mice challenged with fresh M. leprae from passage mice albeit at a slower initial rate. Mice challenged with coculture-extracted bacilli showed evidence of acid-fast bacteria and positive PCR signal for M. leprae. These data support the conclusion that M. leprae can remain viable long-term in environmentally ubiquitous FLA and retain virulence as assessed in the nu/nu mouse model. Additionally, this work supports the idea that M. leprae might be sustained in the environment between hosts in FLA and such residence in FLA may provide a macrophage-like niche contributing to the higher-than-expected rate of leprosy transmission despite a significant decrease in

  8. Mycobacterium leprae phenolglycolipid-1 expressed by engineered M. bovis BCG modulates early interaction with human phagocytes.

    Directory of Open Access Journals (Sweden)

    Guillaume Tabouret

    2010-10-01

    Full Text Available The species-specific phenolic glycolipid 1 (PGL-1 is suspected to play a critical role in the pathogenesis of leprosy, a chronic disease of the skin and peripheral nerves caused by Mycobacterium leprae. Based on studies using the purified compound, PGL-1 was proposed to mediate the tropism of M. leprae for the nervous system and to modulate host immune responses. However, deciphering the biological function of this glycolipid has been hampered by the inability to grow M. leprae in vitro and to genetically engineer this bacterium. Here, we identified the M. leprae genes required for the biosynthesis of the species-specific saccharidic domain of PGL-1 and reprogrammed seven enzymatic steps in M. bovis BCG to make it synthesize and display PGL-1 in the context of an M. leprae-like cell envelope. This recombinant strain provides us with a unique tool to address the key questions of the contribution of PGL-1 in the infection process and to study the underlying molecular mechanisms. We found that PGL-1 production endowed recombinant BCG with an increased capacity to exploit complement receptor 3 (CR3 for efficient invasion of human macrophages and evasion of inflammatory responses. PGL-1 production also promoted bacterial uptake by human dendritic cells and dampened their infection-induced maturation. Our results therefore suggest that M. leprae produces PGL-1 for immune-silent invasion of host phagocytic cells.

  9. Mycobacterium leprae induces NF-κB-dependent transcription repression in human Schwann cells

    International Nuclear Information System (INIS)

    Pereira, Renata M.S.; Calegari-Silva, Teresa Cristina; Hernandez, Maristela O.; Saliba, Alessandra M.; Redner, Paulo; Pessolani, Maria Cristina V.; Sarno, Euzenir N.; Sampaio, Elizabeth P.; Lopes, Ulisses G.

    2005-01-01

    Mycobacterium leprae, the causative agent of leprosy, invades peripheral nerve Schwann cells, resulting in deformities associated with this disease. NF-κB is an important transcription factor involved in the regulation of host immune antimicrobial responses. We aimed in this work to investigate NF-κB signaling pathways in the human ST88-14 Schwannoma cell line infected with M. leprae. Gel shift and supershift assays indicate that two NF-κB dimers, p65/p50 and p50/p50, translocate to the nucleus in Schwann cells treated with lethally irradiated M. leprae. Consistent with p65/p50 and p50/p50 activation, we observed IκB-α degradation and reduction of p105 levels. The nuclear translocation of p50/p50 complex due to M. leprae treatment correlated with repression of NF-κB-driven transcription induced by TNF-α. Moreover, thalidomide inhibited p50 homodimer nuclear translocation induced by M. leprae and consequently rescues Schwann cells from NF-κB-dependent transcriptional repression. Here, we report for the first time that M. leprae induces NF-κB activation in Schwann cells and thalidomide is able to modulate this activation

  10. Mycobacterium leprae-Infected Macrophages Preferentially Primed Regulatory T Cell Responses and Was Associated with Lepromatous Leprosy.

    Directory of Open Access Journals (Sweden)

    Degang Yang

    2016-01-01

    Full Text Available The persistence of Mycobacterium leprae (M. leprae infection is largely dependent on the types of host immune responses being induced. Macrophage, a crucial modulator of innate and adaptive immune responses, could be directly infected by M. leprae. We therefore postulated that M. leprae-infected macrophages might have altered immune functions.Here, we treated monocyte-derived macrophages with live or killed M. leprae, and examined their activation status and antigen presentation. We found that macrophages treated with live M. leprae showed committed M2-like function, with decreased interleukin 1 beta (IL-1beta, IL-6, tumor necrosis factor alpha (TNF-alpha and MHC class II molecule expression and elevated IL-10 and CD163 expression. When incubating with naive T cells, macrophages treated with live M. leprae preferentially primed regulatory T (Treg cell responses with elevated FoxP3 and IL-10 expression, while interferon gamma (IFN-gamma expression and CD8+ T cell cytotoxicity were reduced. Chromium release assay also found that live M. leprae-treated macrophages were more resistant to CD8+ T cell-mediated cytotoxicity than sonicated M. leprae-treated monocytes. Ex vivo studies showed that the phenotype and function of monocytes and macrophages had clear differences between L-lep and T-lep patients, consistent with the in vitro findings.Together, our data demonstrate that M. leprae could utilize infected macrophages by two mechanisms: firstly, M. leprae-infected macrophages preferentially primed Treg but not Th1 or cytotoxic T cell responses; secondly, M. leprae-infected macrophages were more effective at evading CD8+ T cell-mediated cytotoxicity.

  11. Assembly of presynaptic filaments. Factors affecting the assembly of RecA protein onto single-stranded DNA

    DEFF Research Database (Denmark)

    Thresher, RJ; Christiansen, Gunna; Griffith, JD

    1988-01-01

    We have previously shown that the assembly of RecA protein onto single-stranded DNA (ssDNA) facilitated by SSB protein occurs in three steps: (1) rapid binding of SSB protein to the ssDNA; (2) nucleation of RecA protein onto this template; and (3) co-operative polymerization of additional Rec...... assembled onto ssDNA at net rates that varied from 250 to 900 RecA protein monomers per minute, with the rate inversely related to the concentration of SSB protein. Combined sucrose sedimentation and electron microscope analysis established that SSB protein was displaced from the ssDNA during RecA protein...

  12. Novel intein-containing DNA specific primers for rapid identification of Candida glabrata using Real-Time PCR assays.

    Science.gov (United States)

    Kumar, R Satish; Ramesh, S

    2014-12-01

    Candida glabrata is an opportunistic human pathogen known to cause systemic and vaginal candidiasis. Rapid detection of Candida glabrata is indispensable for appropriate selection of antifungal drugs for chemotherapy. The study describes a unique intein-containing DNA fragment for specific detection of C. glabrata. The designed oligonucleotides detected C. glabrata (Ct mean: 24.75 ± 1.1 and Tm: 70.08 ± 0.23°C) in Real-Time PCR assays. The fluorescent signals were negative when the primers were tested for cross-species and cross-genera amplifications. In conclusion, our study recommends a novel primer set for developing a quick identification system which does not require laborious and time-consuming experimentations. Copyright © 2014 Elsevier Masson SAS. All rights reserved.

  13. Distinct DNA repair pathways involving RecA and nonhomologous end joining in Mycobacterium smegmatis.

    OpenAIRE

    Korycka-Machala, M; Brzostek, A; Rozalska, S; Rumijowska-Galewicz, A; Dziedzic, R; Bowater, R; Dziadek, J

    2006-01-01

    Mycobacterium smegmatis was used to study the relationship between DNA repair processes involving RecA and nonhomologous end joining (NHEJ). The effect of gene deletions in recA and/or in two genes involved in NHEJ (ku and ligD) was tested on the ability of bacteria to join breaks in plasmids transformed into them and in their response to chemicals that damage DNA. The results provide in vivo evidence that only NHEJ is required for the repair of noncompatible DNA ends. By contrast, the respon...

  14. Distinct DNA repair pathways involving RecA and nonhomologous end joining in Mycobacterium smegmatis.

    Science.gov (United States)

    Korycka-Machala, Malgorzata; Brzostek, Anna; Rozalska, Sylwia; Rumijowska-Galewicz, Anna; Dziedzic, Renata; Bowater, Richard; Dziadek, Jaroslaw

    2006-05-01

    Mycobacterium smegmatis was used to study the relationship between DNA repair processes involving RecA and nonhomologous end joining (NHEJ). The effect of gene deletions in recA and/or in two genes involved in NHEJ (ku and ligD) was tested on the ability of bacteria to join breaks in plasmids transformed into them and in their response to chemicals that damage DNA. The results provide in vivo evidence that only NHEJ is required for the repair of noncompatible DNA ends. By contrast, the response of mycobacteria to mitomycin C preferentially involved a RecA-dependent pathway.

  15. A multicopy phr-plasmid increases the ultraviolet resistance of a recA strain of Escherichia coli

    International Nuclear Information System (INIS)

    Yamamoto, K.; Satake, M.; Shinagawa, H.

    1984-01-01

    It has been previously reported that the ultraviolet sensitivity of recA strains of Escherichia coli in the dark is suppressed by a plasmid pKY1 which carries the phr gene, suggesting that this is due to a novel effect of photoreactivating enzyme (PRE) of E. coli in the dark. In this work, it is observed that an increase of UV-resistance by pKY1 in the dark is not apparent in strains with a mutation in either uvrA, uvrB, uvrC, lexA, recBC or recF. The sensitivity of recA lexA and recA recBC multiple mutants to UV is suppressed by the plasmid but that of recA uvrA, recA uvrB and recA uvrC is not. Host-cell reactivation of UV-irradiated lambda phage is slightly more efficient in the recA/pKY1 strain compared with the parental recA strain. On the other hand, the recA and recA/pKY1 strains do not differ significantly in the following properties: Hfr recombination, induction of lambda by UV, and mutagenesis. It is suggested that dark repair of PRE is correlated with its capacity of excision repair. (Auth.)

  16. Optimal conditions for decorating outer surface of single-walled carbon nanotubes with RecA proteins

    Science.gov (United States)

    Oura, Shusuke; Umemura, Kazuo

    2016-03-01

    In this study, we estimated the optimal reaction conditions for decorating the outer surface of single-walled carbon nanotubes (SWNTs) with RecA proteins by comparison with hybrids of RecA and single-stranded DNA (ssDNA). To react SWNTs with RecA proteins, we first prepared ssDNA-SWNT hybrids. The heights of the ssDNA-SWNT hybrids increased as the amount of RecA used in the reaction increased, as determined from atomic force microscopy images. We further confirmed the increasing adsorption of RecA proteins onto ssDNA on SWNT surfaces by agarose gel electrophoresis. These results suggest that the combination of RecA proteins and ssDNA-SWNT hybrids forms RecA-ssDNA-SWNT hybrids. We also successfully controlled the amount of RecA adsorbed on the ssDNA-SWNT hybrids. Our results thus indicate the optimized reaction conditions for decorating the outer surface of SWNTs with RecA proteins, which is the key to the development of novel biosensors and nanomaterial-based bioelectronics.

  17. Characterization of murine B-cell epitopes on the Mycobacterium leprae proline-rich antigen by use of synthetic peptides

    NARCIS (Netherlands)

    Klatser, P. R.; de Wit, M. Y.; Kolk, A. H.; Hartskeerl, R. A.

    1991-01-01

    Using synthetic peptides representing overlapping sequences of the 100-amino-acid-long N-terminal region of the proline-rich antigen of Mycobacterium leprae (PRA), we have mapped the epitopes in the primary structure of PRA recognized by four monoclonal antibodies. The M. leprae-specific monoclonal

  18. Vigilancia de la lepra en situaciones de baja prevalencia

    Directory of Open Access Journals (Sweden)

    C. Edilberto González Ochoa

    2001-02-01

    Full Text Available La mayoría de los países latinoamericanos han logrado reducir la prevalencia de la lepra a menos de 1 caso por cada 10 000 habitantes. En estos países, la etapa siguiente es eliminar la enfermedad en el ámbito subnacional, en los territorios que tienen tasas mayores de 1 caso por 10 000. Elementos como la transición demográfica, la existencia de áreas con elevada transmisión y la necesidad de emplear indicadores más sensibles obligan a modificar las estrategias básicas, fortalecer los sistemas de vigilancia y reorientar recursos según sea necesario. Es importante renovar el empleo de tácticas como la identificación de las áreas críticas, las intervenciones diferenciadas, la concentración de indicadores y la conjugación de la vigilancia pasiva y activa. Esto puede formularse rediseñando los sistemas de vigilancia para integrar los componentes clínico, de laboratorio, de investigación epidemiológica y de suministros. Los resultados del proceso deben aportar un conjunto mínimo de indicadores que permitan monitorear y evaluar la efectividad y la eficiencia del plan de acción para la etapa posteliminación.

  19. RecA bundles mediate homology pairing between distant sisters during DNA break repair

    Science.gov (United States)

    Lesterlin, Christian; Ball, Graeme; Schermelleh, Lothar; Sherratt, David J.

    2014-02-01

    DNA double-strand break (DSB) repair by homologous recombination has evolved to maintain genetic integrity in all organisms. Although many reactions that occur during homologous recombination are known, it is unclear where, when and how they occur in cells. Here, by using conventional and super-resolution microscopy, we describe the progression of DSB repair in live Escherichia coli. Specifically, we investigate whether homologous recombination can occur efficiently between distant sister loci that have segregated to opposite halves of an E. coli cell. We show that a site-specific DSB in one sister can be repaired efficiently using distant sister homology. After RecBCD processing of the DSB, RecA is recruited to the cut locus, where it nucleates into a bundle that contains many more RecA molecules than can associate with the two single-stranded DNA regions that form at the DSB. Mature bundles extend along the long axis of the cell, in the space between the bulk nucleoid and the inner membrane. Bundle formation is followed by pairing, in which the two ends of the cut locus relocate at the periphery of the nucleoid and together move rapidly towards the homology of the uncut sister. After sister locus pairing, RecA bundles disassemble and proteins that act late in homologous recombination are recruited to give viable recombinants 1-2-generation-time equivalents after formation of the initial DSB. Mutated RecA proteins that do not form bundles are defective in sister pairing and in DSB-induced repair. This work reveals an unanticipated role of RecA bundles in channelling the movement of the DNA DSB ends, thereby facilitating the long-range homology search that occurs before the strand invasion and transfer reactions.

  20. Lepra: enfermedad milenaria y actual = Leprosy: an ancient and present-day disease

    Directory of Open Access Journals (Sweden)

    Cardona Castro, Nora María

    2011-03-01

    Full Text Available El desconocimiento de la lepra es común en la población general al igual que entre los médicos y el personal de la salud. Se cree que esta enfermedad ya no existe; tal vez su imagen bíblica y milenaria refuerce la idea de su eliminación. Sin embargo, la lepra continúa siendo un problema de salud pública en varios países; entre los más afectados están India y Brasil. Después del inicio de la poliquimioterapia (PQT en la novena década del siglo XX la prevalencia de la lepra disminuyó considerablemente pero no ocurrió lo mismo con la incidencia, lo que se atribuye al poco impacto de dicho tratamiento sobre el control de la transmisión y a la existencia de un reservorio aún no identificado con exactitud. Los convivientes de los leprosos tienen alto riesgo de sufrir la enfermedad en cualquier momento de la vida, pero hasta ahora no se ha podido determinar cuáles convivientes infectados desarrollarán la enfermedad. En Colombia se informan de 400 a 550 casos de lepra cada año, lo cual sugiere que la transmisión del Mycobacterium leprae continúa a pesar de que el país está considerado en la fase de poseliminación.Este artículo presenta una revisión histórica de la lepra desde los primeros informes disponibles hasta los avances moleculares más recientes. Incluye cómo ha evolucionado la comprensión de la enfermedad, su caracterización clínica, las medidas de control y saneamiento, el tratamiento y la epidemiología.

  1. Detection of Lsr2 gene of Mycobacterium leprae in nasal mucus

    Directory of Open Access Journals (Sweden)

    Luiz Antonio Custodio

    2012-06-01

    Full Text Available In the present study, nasal mucus from patients with leprosy were analyzed by PCR using specific primers for Lsr2 gene of Mycobacterium leprae. The presence of Lsr2 gene in the nasal mucus was detected in 25.80% of patients with paucibacillari leprosy, and 23.07% of contacts. Despite the absence of clinical features in the contact individuals, it was possible to detect the presence of Lsr2 gene in the nasal mucus of these individuals. Therefore, PCR detection of M. leprae targeting Lsr2 gene using nasal mucus samples could contribute to early diagnosis of leprosy.

  2. The majority of inducible DNA repair genes in Mycobacterium tuberculosis are induced independently of RecA.

    Science.gov (United States)

    Rand, Lucinda; Hinds, Jason; Springer, Burkhard; Sander, Peter; Buxton, Roger S; Davis, Elaine O

    2003-11-01

    In many species of bacteria most inducible DNA repair genes are regulated by LexA homologues and are dependent on RecA for induction. We have shown previously by analysing the induction of recA that two mechanisms for the induction of gene expression following DNA damage exist in Mycobacterium tuberculosis. Whereas one of these depends on RecA and LexA in the classical way, the other mechanism is independent of both of these proteins and induction occurs in the absence of RecA. Here we investigate the generality of each of these mechanisms by analysing the global response to DNA damage in both wild-type M. tuberculosis and a recA deletion strain of M. tuberculosis using microarrays. This revealed that the majority of the genes that were induced remained inducible in the recA mutant stain. Of particular note most of the inducible genes with known or predicted functions in DNA repair did not depend on recA for induction. Amongst these are genes involved in nucleotide excision repair, base excision repair, damage reversal and recombination. Thus, it appears that this novel mechanism of gene regulation is important for DNA repair in M. tuberculosis.

  3. Purification and characterization of a 36 kDa antigen of Mycobacterium leprae

    NARCIS (Netherlands)

    de Wit, M. Y.; Klatser, P. R.

    1988-01-01

    A 36 kDa antigen of Mycobacterium leprae was purified by phenol biphasic partition followed by preparative SDS-PAGE. The purified antigen appeared as a single band in SDS-PAGE and eluted as a single peak in ion-exchange chromatography. The antigen comprised epitopes which were cross-reactive with M.

  4. Polymerase chain reaction for detection of Mycobacterium leprae in nasal swab specimens

    NARCIS (Netherlands)

    de Wit, M. Y.; Douglas, J. T.; McFadden, J.; Klatser, P. R.

    1993-01-01

    The polymerase chain reaction based on the selective amplification of a 531-bp fragment of the gene encoding the proline-rich antigen of Mycobacterium leprae was applied to nasal swab specimens from leprosy patients, occupational contacts, and endemic and nonendemic controls. To prevent

  5. Detection and Strain Typing of Ancient Mycobacterium leprae from a Medieval Leprosy Hospital

    Science.gov (United States)

    Taylor, G. Michael; Tucker, Katie; Butler, Rachel; Pike, Alistair W. G.; Lewis, Jamie; Roffey, Simon; Marter, Philip; Lee, Oona Y-C; Wu, Houdini H. T.; Minnikin, David E.; Besra, Gurdyal S.; Singh, Pushpendra; Cole, Stewart T.; Stewart, Graham R.

    2013-01-01

    Nine burials excavated from the Magdalen Hill Archaeological Research Project (MHARP) in Winchester, UK, showing skeletal signs of lepromatous leprosy (LL) have been studied using a multidisciplinary approach including osteological, geochemical and biomolecular techniques. DNA from Mycobacterium leprae was amplified from all nine skeletons but not from control skeletons devoid of indicative pathology. In several specimens we corroborated the identification of M. leprae with detection of mycolic acids specific to the cell wall of M. leprae and persistent in the skeletal samples. In five cases, the preservation of the material allowed detailed genotyping using single-nucleotide polymorphism (SNP) and multiple locus variable number tandem repeat analysis (MLVA). Three of the five cases proved to be infected with SNP type 3I-1, ancestral to contemporary M. leprae isolates found in southern states of America and likely carried by European migrants. From the remaining two burials we identified, for the first time in the British Isles, the occurrence of SNP type 2F. Stable isotope analysis conducted on tooth enamel taken from two of the type 3I-1 and one of the type 2F remains revealed that all three individuals had probably spent their formative years in the Winchester area. Previously, type 2F has been implicated as the precursor strain that migrated from the Middle East to India and South-East Asia, subsequently evolving to type 1 strains. Thus we show that type 2F had also spread westwards to Britain by the early medieval period. PMID:23638071

  6. Mutation analysis of the Mycobacterium leprae folP1 gene and dapsone resistance.

    Science.gov (United States)

    Nakata, Noboru; Kai, Masanori; Makino, Masahiko

    2011-02-01

    Diaminodiphenylsulfone (dapsone) has long been used as a first-line drug worldwide for the treatment of leprosy. Diagnosis for dapsone resistance of Mycobacterium leprae by DNA tests would be of great clinical value, but the relationship between the nucleotide substitutions and susceptibility to dapsone must be clarified before use. In this study, we constructed recombinant strains of cultivable Mycobacterium smegmatis carrying the M. leprae folP1 gene with or without a point mutation, disrupting their own folP gene on the chromosome. Dapsone susceptibilities of the recombinant bacteria were measured to examine influence of the mutations. Dapsone MICs for most of the strains with mutations at codon 53 or 55 of M. leprae folP1 were 2 to 16 times as high as the MIC for the strain with the wild-type folP1 sequence, but mutations that changed Thr to Ser at codon 53 showed somewhat lower MIC values than the wild-type sequence. Strains with mutations at codon 48 or 54 showed levels of susceptibility to dapsone comparable to the susceptibility of the strain with the wild-type sequence. This study confirmed that point mutations at codon 53 or 55 of the M. leprae folP1 gene result in dapsone resistance.

  7. Mutation Analysis of the Mycobacterium leprae folP1 Gene and Dapsone Resistance▿

    Science.gov (United States)

    Nakata, Noboru; Kai, Masanori; Makino, Masahiko

    2011-01-01

    Diaminodiphenylsulfone (dapsone) has long been used as a first-line drug worldwide for the treatment of leprosy. Diagnosis for dapsone resistance of Mycobacterium leprae by DNA tests would be of great clinical value, but the relationship between the nucleotide substitutions and susceptibility to dapsone must be clarified before use. In this study, we constructed recombinant strains of cultivable Mycobacterium smegmatis carrying the M. leprae folP1 gene with or without a point mutation, disrupting their own folP gene on the chromosome. Dapsone susceptibilities of the recombinant bacteria were measured to examine influence of the mutations. Dapsone MICs for most of the strains with mutations at codon 53 or 55 of M. leprae folP1 were 2 to 16 times as high as the MIC for the strain with the wild-type folP1 sequence, but mutations that changed Thr to Ser at codon 53 showed somewhat lower MIC values than the wild-type sequence. Strains with mutations at codon 48 or 54 showed levels of susceptibility to dapsone comparable to the susceptibility of the strain with the wild-type sequence. This study confirmed that point mutations at codon 53 or 55 of the M. leprae folP1 gene result in dapsone resistance. PMID:21115799

  8. Detection and strain typing of ancient Mycobacterium leprae from a medieval leprosy hospital.

    Directory of Open Access Journals (Sweden)

    G Michael Taylor

    Full Text Available Nine burials excavated from the Magdalen Hill Archaeological Research Project (MHARP in Winchester, UK, showing skeletal signs of lepromatous leprosy (LL have been studied using a multidisciplinary approach including osteological, geochemical and biomolecular techniques. DNA from Mycobacterium leprae was amplified from all nine skeletons but not from control skeletons devoid of indicative pathology. In several specimens we corroborated the identification of M. leprae with detection of mycolic acids specific to the cell wall of M. leprae and persistent in the skeletal samples. In five cases, the preservation of the material allowed detailed genotyping using single-nucleotide polymorphism (SNP and multiple locus variable number tandem repeat analysis (MLVA. Three of the five cases proved to be infected with SNP type 3I-1, ancestral to contemporary M. leprae isolates found in southern states of America and likely carried by European migrants. From the remaining two burials we identified, for the first time in the British Isles, the occurrence of SNP type 2F. Stable isotope analysis conducted on tooth enamel taken from two of the type 3I-1 and one of the type 2F remains revealed that all three individuals had probably spent their formative years in the Winchester area. Previously, type 2F has been implicated as the precursor strain that migrated from the Middle East to India and South-East Asia, subsequently evolving to type 1 strains. Thus we show that type 2F had also spread westwards to Britain by the early medieval period.

  9. Toxoide de la difteria en el tratamiento de la lepra (1)

    OpenAIRE

    Chala H., J. Ignacio; Lleras Restrepo, Federico

    2011-01-01

    En abril de 1939 el doctor D. R. Collier de "Cheingmai Leper Asylum", Thailand, informó sobre los óptimos resultados obtenidos en el tratamiento de la lepra por medio de la "antitoxina" y "toxoide" de la difteria. Las investigaciones las llevó a cabo en un grupo de enfermos de ese leprosorio.

  10. Toxoide de la difteria en el tratamiento de la lepra (1

    Directory of Open Access Journals (Sweden)

    J. Ignacio Chala H.

    1943-09-01

    Full Text Available En abril de 1939 el doctor D. R. Collier de "Cheingmai Leper Asylum", Thailand, informó sobre los óptimos resultados obtenidos en el tratamiento de la lepra por medio de la "antitoxina" y "toxoide" de la difteria. Las investigaciones las llevó a cabo en un grupo de enfermos de ese leprosorio.

  11. Electron microscopic visualization of the RecA protein-mediated pairing and branch migration phases of DNA strand exchange

    DEFF Research Database (Denmark)

    Register, JC; Christiansen, Gunna; Griffith, J

    1987-01-01

    The RecA protein of Escherichia coli will drive the pairing and exchange of strands between homologous DNA molecules in a reaction stimulated by single-stranded binding protein. Here, reactions utilizing three homologous DNA pairs which can undergo both paranemic and plectonemic joining were...... indistinguishable from paranemic joints; (ii) complexes which appeared to be joined both paranemically and plectonemically were present in these reactions in roughly equal numbers; and (iii) in complexes undergoing strand exchange, both DNA partners were often enveloped within a RecA protein filament consisting...... of hundreds of RecA protein monomers and several kilobases of DNA. These observations suggest that, following RecA protein-ssDNA filament formation, strand exchange proceeds by a pathway that can be divided structurally into three phases: pairing, envelopment/exchange, and release of the products....

  12. Sequence homology and expression profile of genes associated with DNA repair pathways in Mycobacterium leprae.

    Science.gov (United States)

    Sharma, Mukul; Vedithi, Sundeep Chaitanya; Das, Madhusmita; Roy, Anindya; Ebenezer, Mannam

    2017-01-01

    Survival of Mycobacterium leprae, the causative bacteria for leprosy, in the human host is dependent to an extent on the ways in which its genome integrity is retained. DNA repair mechanisms protect bacterial DNA from damage induced by various stress factors. The current study is aimed at understanding the sequence and functional annotation of DNA repair genes in M. leprae. T he genome of M. leprae was annotated using sequence alignment tools to identify DNA repair genes that have homologs in Mycobacterium tuberculosis and Escherichia coli. A set of 96 genes known to be involved in DNA repair mechanisms in E. coli and Mycobacteriaceae were chosen as a reference. Among these, 61 were identified in M. leprae based on sequence similarity and domain architecture. The 61 were classified into 36 characterized gene products (59%), 11 hypothetical proteins (18%), and 14 pseudogenes (23%). All these genes have homologs in M. tuberculosis and 49 (80.32%) in E. coli. A set of 12 genes which are absent in E. coli were present in M. leprae and in Mycobacteriaceae. These 61 genes were further investigated for their expression profiles in the whole transcriptome microarray data of M. leprae which was obtained from the signal intensities of 60bp probes, tiling the entire genome with 10bp overlaps. It was noted that transcripts corresponding to all the 61 genes were identified in the transcriptome data with varying expression levels ranging from 0.18 to 2.47 fold (normalized with 16SrRNA). The mRNA expression levels of a representative set of seven genes ( four annotated and three hypothetical protein coding genes) were analyzed using quantitative Polymerase Chain Reaction (qPCR) assays with RNA extracted from skin biopsies of 10 newly diagnosed, untreated leprosy cases. It was noted that RNA expression levels were higher for genes involved in homologous recombination whereas the genes with a low level of expression are involved in the direct repair pathway. This study provided

  13. Sequence homology and expression profile of genes associated with dna repair pathways in Mycobacterium leprae

    Directory of Open Access Journals (Sweden)

    Mukul Sharma

    2017-01-01

    Full Text Available Background: Survival of Mycobacterium leprae, the causative bacteria for leprosy, in the human host is dependent to an extent on the ways in which its genome integrity is retained. DNA repair mechanisms protect bacterial DNA from damage induced by various stress factors. The current study is aimed at understanding the sequence and functional annotation of DNA repair genes in M. leprae. Methods: T he genome of M. leprae was annotated using sequence alignment tools to identify DNA repair genes that have homologs in Mycobacterium tuberculosis and Escherichia coli. A set of 96 genes known to be involved in DNA repair mechanisms in E. coli and Mycobacteriaceae were chosen as a reference. Among these, 61 were identified in M. leprae based on sequence similarity and domain architecture. The 61 were classified into 36 characterized gene products (59%, 11 hypothetical proteins (18%, and 14 pseudogenes (23%. All these genes have homologs in M. tuberculosis and 49 (80.32% in E. coli. A set of 12 genes which are absent in E. coli were present in M. leprae and in Mycobacteriaceae. These 61 genes were further investigated for their expression profiles in the whole transcriptome microarray data of M. leprae which was obtained from the signal intensities of 60bp probes, tiling the entire genome with 10bp overlaps. Results: It was noted that transcripts corresponding to all the 61 genes were identified in the transcriptome data with varying expression levels ranging from 0.18 to 2.47 fold (normalized with 16SrRNA. The mRNA expression levels of a representative set of seven genes ( four annotated and three hypothetical protein coding genes were analyzed using quantitative Polymerase Chain Reaction (qPCR assays with RNA extracted from skin biopsies of 10 newly diagnosed, untreated leprosy cases. It was noted that RNA expression levels were higher for genes involved in homologous recombination whereas the genes with a low level of expression are involved in the

  14. Purification and characterization of the RecA protein from Neisseria gonorrhoeae.

    Directory of Open Access Journals (Sweden)

    Elizabeth A Stohl

    2011-02-01

    Full Text Available The strict human pathogen Neisseria gonorrhoeae is the only causative agent of the sexually transmitted infection gonorrhea. The recA gene from N. gonorrhoeae is essential for DNA repair, natural DNA transformation, and pilin antigenic variation, all processes that are important for the pathogenesis and persistence of N. gonorrhoeae in the human population. To understand the biochemical features of N. gonorrhoeae RecA (RecA(Ng, we overexpressed and purified the RecA(Ng and SSB(Ng proteins and compared their activities to those of the well-characterized E. coli RecA and SSB proteins in vitro. We observed that RecA(Ng promoted more strand exchange at early time points than RecA(Ec through DNA homologous substrates, and exhibited the highest ATPase activity of any RecA protein characterized to date. Further analysis of this robust ATPase activity revealed that RecA(Ng is more efficient at displacing SSB from ssDNA and that RecA(Ng shows higher ATPase activity during strand exchange than RecA(Ec. Using substrates created to mimic the cellular processes of DNA transformation and pilin antigenic variation we observed that RecA(Ec catalyzed more strand exchange through a 100 bp heterologous insert, but that RecA(Ng catalyzed more strand exchange through regions of microheterology. Together, these data suggest that the processes of ATP hydrolysis and DNA strand exchange may be coupled differently in RecA(Ng than in RecA(Ec. This difference may explain the unusually high ATPase activity observed for RecA(Ng with the strand exchange activity between RecA(Ng and RecA(Ec being more similar.

  15. Investigating direct interaction between Escherichia coli topoisomerase I and RecA

    Science.gov (United States)

    Darici, Yesim; Tse-Dinh, Yuk-Ching

    2016-01-01

    Protein-protein interactions are of special importance in cellular processes, including replication, transcription, recombination, and repair. Escherichia coli topoisomerase I (EcTOP1) is primarily involved in the relaxation of negative DNA supercoiling. E. coli RecA, the key protein for homologous recombination and SOS DNA-damage response, has been shown to stimulate the relaxation activity of EcTOP1. The evidence for their direct protein-protein interaction has not been previously established. We report here the direct physical interaction between E. coli RecA and topoisomerase I. We demonstrated the RecA-topoisomerase I interaction via pull-down assays, and surface plasmon resonance measurements. Molecular docking supports the observation that the interaction involves the topoisomerase I N-terminal domains that form the active site. Our results from pull-down assays showed that ATP, although not required, enhances the RecA-EcTOP1 interaction. We propose that E. coli RecA physically interacts with topoisomerase I to modulate the chromosomal DNA supercoiling. PMID:27001450

  16. Factores asociados a recaídas por tuberculosis en Lima este - Perú

    Directory of Open Access Journals (Sweden)

    María Ríos Hipólito

    2002-01-01

    Full Text Available Objetivo: Determinar los factores de riesgo asociados a recaídas por tuberculosis en Lima Este - Perú, entre marzo y diciembre del 2000. Materiales y métodos: estudio caso-control. Se definió a los casos (184 como los pacientes que recibieron tratamiento con el esquema I alguna vez, egresaron como curados y volvieron a presentar otro episodio de tuberculosis BK positivo durante 1999. Los controles (368 fueron los pacientes nuevos con tuberculosis BK positivo tratados en 1998 que no recayeron. Resultados: se asociaron significativamente a las recaídas el sexo masculino, la edad mayor de 50 años, el consumo de drogas, la residencia en un área urbana, el hacinamiento, la percepción errada de la enfermedad (PEE y la desocupación, no así el contacto con un paciente tuberculoso. Luego del análisis multivariado, sólo se asociaron el área urbana, el hacinamiento, la PEE y el tratamiento irregular. Conclusiones: la residencia en un área urbana, el hacinamiento, la PEE y la irregularidad en el tratamiento son factores asociados significativamente a recaídas en pacientes con TBC pulmonar BK(+ de Lima Este, Perú.

  17. RecA Inhibitors Potentiate Antibiotic Activity and Block Evolution of Antibiotic Resistance.

    Science.gov (United States)

    Alam, Md Kausar; Alhhazmi, Areej; DeCoteau, John F; Luo, Yu; Geyer, C Ronald

    2016-03-17

    Antibiotic resistance arises from the maintenance of resistance mutations or genes acquired from the acquisition of adaptive de novo mutations or the transfer of resistance genes. Antibiotic resistance is acquired in response to antibiotic therapy by activating SOS-mediated DNA repair and mutagenesis and horizontal gene transfer pathways. Initiation of the SOS pathway promotes activation of RecA, inactivation of LexA repressor, and induction of SOS genes. Here, we have identified and characterized phthalocyanine tetrasulfonic acid RecA inhibitors that block antibiotic-induced activation of the SOS response. These inhibitors potentiate the activity of bactericidal antibiotics, including members of the quinolone, β-lactam, and aminoglycoside families in both Gram-negative and Gram-positive bacteria. They reduce the ability of bacteria to acquire antibiotic resistance mutations and to transfer mobile genetic elements conferring resistance. This study highlights the advantage of including RecA inhibitors in bactericidal antibiotic therapies and provides a new strategy for prolonging antibiotic shelf life. Copyright © 2016 Elsevier Ltd. All rights reserved.

  18. Cell survival, UV-reactivation and induction of prophage lambda in Escherichia coli K12 overproducing RecA protein

    International Nuclear Information System (INIS)

    Quillardet, P.; Moreau, P.L.; Devoret, R.; Ginsburg, H.; Mount, D.W.

    1982-01-01

    The effect of the cellular level of RecA protein on the ability of E. coli K12 bacteria to (I) survive UV-irradiation (II) promote UV-reactivation of UV-damaged phage lambda (III) induce prophage lambda was determined in bacterial mutants with discrete increasing levels of RecA protein. The various levels of RecA protein were obtained by combining lexA and recA alleles. Except for the double mutant lexA3 recAo98, whose repair ability was 25% less than that observed in wild type bacteria, bacterial survival was proportional to the level of ReCA protein measured after 90 min of incubation. In lexA3 recAo98 bacteria, RecA protein, at a constitutive high basal level, failed to compensate totally for the lack of LexA repressor cleavage; UV-reactivation of UV-damaged phage lambda was not restored; yet, prophage lambda was induced with 35% efficiency. Efficient UV-induction of prophage lambda is linked to the induction of lexA-controlled host processes that repair the UV-damaged prophage. (orig.)

  19. [Cloning of the recA gene of the propionic acid bacterium Propionibacterium shermanii in Escherichia coli cells].

    Science.gov (United States)

    Pankova, S V; Avilev, S K; Tarasov, V A; Ogarkova, O A

    1993-05-01

    Propionibacterium are producers of vitamin B-12 and organic acids and are of importance in national economy. Genetics of this organism was studied insufficiently. We constructed the genomic library of Propionibacterium shermanii in cells of Escherichia coli using the plasmid vector pVZ361 and identified recA gene. The vector gives a chance for direct selection of Str-resistant clones containing an insert in BamHI site. The recombinant plasmid carrying the recA gene of P. shermanii was isolated from the genomic library using complementation in E. coli. Strains E. coli C600 and HB101 were transformed by hybrid plasmids, and UV-light-resistant clones were identified. The clones were purified and subjected to treatment with 4-NQO and MMS. Diagrams reflecting survival dependence of the bacteria carrying recombinant plasmids and lacking them on the mutagen concentration and UV-light dose clearly confirmed functioning of P. shermanii recA gene in E. coli cells. The insert with recA gene underwent restriction analysis. The 1.7 kb fragment with recA gene was then transferred to pBI101 plasmid and the resultant recombinant plasmid was used in the SOS test. The mutagens (MMS, 4-NHQ) and UV-light induced the SOS response in E. coli HB101 (recA) carrying the recombinant plasmid.

  20. "Batallas contra la Lepra: Estado, Medicina y Ciencia en Colombia"

    Directory of Open Access Journals (Sweden)

    Carlos Lleras de la Fuente

    2003-03-01

    Full Text Available

    Hace pocos meses tuve el gusto de conocer a Diana Obregón Torres, de quien ya había oído hablar con ocasión de haber ella ganado el premio de Ciencias Sociales y Humanas de la Fundación
    Alejandro Ángel Escobar y de un reportaje que una amiga de mi hija le había hecho.
    Se me presentó en algún evento al cual ambos habíamos concurrido y tuve de ella una primera impresión equivocada: la de una persona suave, estudíosa, de bajo perfíl. Es este tipo de errores que
    lleva a que se desbaraten tantos matrimonios.
    La verdadera Diana es una investigadora rigurosa, erudita, disciplinada, que emite sus opiniones sobre todo y todos sin recato alguno y con una dureza que en muchos casos resulta urticante.
    Esa figura surgió del magnífico libro que el Fondo Editorial Universitario Eafit y el Banco de la República editaron en julio de este año, y cuyas 378 págínas de texto apasionante leí en un par de días, sin contar el vistazo que dí a las 37 páginas de la bibliografía las cuales, además y en notas de pie de página, respaldan la seriedad de su investigación.
    La lectura del líbro es, para un devorad'or de novelas de espionaje, de terror y policiacas, una triple ración de ese suspenso que, ya cerca del amanecer, no nos deja parar. Todos los elementos están en él
    presentes: el criminal es la lepra, que nos transporta al antíguo y al nuevo testamento; ha estado causando terror por varios milenios y nadie ha podido encarcelarla ni, por supuesto, aplicarle la tan desacreditada pena de muerte.
    Hoy día sigue tan campante y de vez en cuando se hace sentír desafíando las sulfonas y el aceite de chaulmugra. Es una enfermedad infecciosa, sin duda, pero el bacilo llamado "de Hansen" se ha defendido con éxito de los bacteriólogos quienes no han logrado producir la vacuna que permitiría erradicar este mal de la faz de la tierra...

  1. Morfologia do Mycobacterium leprae hominis e do M. leprae muris: estudo baseado na microscopia electrônica e de contraste de fases Morphology of Mycobacterium leprae hominis and M. leprae muris based on electron and phases contrast microscopy

    Directory of Open Access Journals (Sweden)

    H. C. de Souza-Araújo

    1955-12-01

    Full Text Available Hansen's Bacillus: By electron microscopy this bacillus shows membrane and halo, this being more visible when sorrounding the globi or bundles of bacilli; shows, also, free granules of various sizes which were before considered as dust of the dyes; shows external granules bound with the membrane and some times branching. By phases contrast microscopy examining leproma suspensions and subcataneous lymph at 400 x we saw many free granules with intense rotatory movement; granulated bacilli with screw, skip or stroke motion, producing slow progressive motion. All such elementes are surrounded by a halo, corresponding to the classical gloea. By a patient and delayed examination we were able to see that the internal granules are motile and help the progression of the bacilli, giving the impression that the cytoplasm is liquid. By a lasting observation we could see the larger granules form prolapse, like a pseudopode and abandon the bacilli and going in very rapid rotatory movement. There are branched bacilli; there are pedunculated fred granules like comets. The addition of a drop of formol at the preparation stops all movements. Stefansky's Bacillus: Repeated examination by RCA electron microscope, type EMU-25 of fresh suspensions of rat lepromas, led us to confirm the close relationship between human and murine leprosy agents. We examined also material from carabo (Lepra bubalorum from Java, but due to fixation, the material was unsuitable for comparative studies. The Stefansky's bacilli showed also emmbranes and halos, internal or external granules (smaller than those of Hansen's bacillus. The bacilli shaded by chromium look thicker and shorter than those of Hansen. Due to electron bombardment both, Hansen's and Stefansky's baccilli suffer considerable alterations in their structure, showing black barrs of chromatin condensation at their extremities as also in their centers. By phase microscopy the Stefansky's bacilli showed elements with 1, 2

  2. Rapid, radiolabeled-microculture method that uses macrophages for in vitro evaluation of Mycobacterium leprae viability and drug susceptibility.

    Science.gov (United States)

    Mittal, A; Sathish, M; Seshadri, P S; Nath, I

    1983-04-01

    This paper describes a microculture rapid assay using radiolabeling and mouse macrophages to determine the viability and the drug susceptibility or resistance of Mycobacterium leprae. Comparison of M. leprae resident macrophage cultures maintained in 96-well flat-bottomed plates showed results for viability and susceptibility or resistance to dapsone that were similar to results for concurrent cultures in Leighton tubes with greater numbers of bacilli and macrophages.

  3. Rapid, Radiolabeled-Microculture Method That Uses Macrophages for In Vitro Evaluation of Mycobacterium leprae Viability and Drug Susceptibility

    OpenAIRE

    Mittal, A.; Sathish, M.; Seshadri, P. S.; Nath, Indira

    1983-01-01

    This paper describes a microculture rapid assay using radiolabeling and mouse macrophages to determine the viability and the drug susceptibility or resistance of Mycobacterium leprae. Comparison of M. leprae resident macrophage cultures maintained in 96-well flat-bottomed plates showed results for viability and susceptibility or resistance to dapsone that were similar to results for concurrent cultures in Leighton tubes with greater numbers of bacilli and macrophages.

  4. qPCR detection of Mycobacterium leprae in biopsies and slit skin smear of different leprosy clinical forms

    OpenAIRE

    Michelle de Campos Soriani Azevedo; Natália Mortari Ramuno; Luciana Raquel Vincenzi Fachin; Mônica Tassa; Patrícia Sammarco Rosa; Andrea de Faria Fernandes Belone; Suzana Madeira Diório; Cleverson Teixeira Soares; Gustavo Pompermaier Garlet; Ana Paula Favaro Trombone

    2017-01-01

    Abstract Leprosy, whose etiological agent is Mycobacterium leprae, is a chronic infectious disease that mainly affects the skin and peripheral nervous system. The diagnosis of leprosy is based on clinical evaluation, whereas histopathological analysis and bacilloscopy are complementary diagnostic tools. Quantitative PCR (qPCR), a current useful tool for diagnosis of infectious diseases, has been used to detect several pathogens including Mycobacterium leprae. The validation of this technique ...

  5. Extraction and detection of Mycobacterium leprae DNA from ZNCF-stained skin smear slides for better identification of negative skin smears

    Directory of Open Access Journals (Sweden)

    Kamble R

    2010-01-01

    Full Text Available Abstract Identification of Mycobacterium leprae, which causes leprosy, is done by Ziehl Neelsen Carbol Fuchsin (ZNCF stained slit skin smear microscopy that aids in the diagnosis and quantification of approximate bacterial load carried by the patient. We attempted M. leprae DNA extraction from 46 stained slit skin smear negative slides, using Proteinase K and SDS lysis, followed by ethanol precipitation. M. leprae specific primers (16SrRNA were used for PCR-based amplification of DNA. We could detect M. leprae DNA in 15 (32.6% samples. The method can be useful in the diagnosis of apparently slit skin smear negative leprosy cases.

  6. Deinococcus radiodurans RecA nucleoprotein filaments characterized at the single-molecule level with optical tweezers

    Energy Technology Data Exchange (ETDEWEB)

    Pobegalov, Georgii, E-mail: george.pobegalov@nanobio.spbstu.ru [Peter the Great St.Petersburg Polytechnic University, Saint-Petersburg 195251 (Russian Federation); Cherevatenko, Galina; Alekseev, Aleksandr; Sabantsev, Anton; Kovaleva, Oksana; Vedyaykin, Alexey; Morozova, Natalia [Peter the Great St.Petersburg Polytechnic University, Saint-Petersburg 195251 (Russian Federation); Baitin, Dmitrii [Peter the Great St.Petersburg Polytechnic University, Saint-Petersburg 195251 (Russian Federation); Petersburg Nuclear Physics Institute, NRC Kurchatov Institute, Gatchina 188300 (Russian Federation); Khodorkovskii, Mikhail [Peter the Great St.Petersburg Polytechnic University, Saint-Petersburg 195251 (Russian Federation)

    2015-10-23

    Deinococcus radiodurans can survive extreme doses of ionizing radiation due to the very efficient DNA repair mechanisms that are able to cope even with hundreds of double-strand breaks. RecA, the critical protein of homologous recombination in bacteria, is one of the key components of the DNA-repair system. Repair of double-strand breaks requires RecA binding to DNA and assembly of the RecA nucleoprotein helical filaments. The Escherichia coli RecA protein (EcRecA) and its interactions with DNA have been extensively studied using various approaches including single-molecule techniques, while the D. radiodurans RecA (DrRecA) remains much less characterized. However, DrRecA shows some remarkable differences from E. coli homolog. Here we combine microfluidics and single-molecule DNA manipulation with optical tweezers to follow the binding of DrRecA to long double-stranded DNA molecules and probe the mechanical properties of DrRecA nucleoprotein filaments at physiological pH. Our data provide a direct comparison of DrRecA and EcRecA binding to double-stranded DNA under identical conditions. We report a significantly faster filaments assembly as well as lower values of persistence length and contour length for DrRecA nucleoprotein filaments compared to EcRecA. Our results support the existing model of DrRecA forming more frequent and less continuous filaments relative to those of EcRecA. - Highlights: • We investigate Deinococcus radiodurans RecA interactions with long double-stranded DNA at the single-molecule level. • At physiological pH D. radiodurans RecA forms nucleoprotein filaments significantly faster relative to Escherichia coli RecA. • D. radiodurans RecA-dsDNA nucleoprotein filaments are more flexible and slightly shorter compared to those of E. coli RecA.

  7. Polimorfismos en el gen promotor de IL-10 en una muestra de pacientes colombianos con lepra

    Directory of Open Access Journals (Sweden)

    Nora Cardona-Castro

    2012-03-01

    Conclusiones. El haplotipo que encontramos asociado con lepra, -1082A-819C-592C/-1082A-819C-592C, se ha relacionado con baja producción de IL-10. Funcionalmente, esta baja producción de IL-10 puede tener consecuencias en la respuesta inmunitaria, además de implicaciones clínicas. Se han reportado diferentes haplotipos de IL-10 como marcadores de vulnerabilidad y resistencia de lepra en otras poblaciones, lo cual sugiere que las diferencias en la distribución de diversos polimorfismos del gen de IL-10 entre grupos étnicos, es un factor importante al determinar la asociación entre enfermedad y genes.   DOI: http://dx.doi.org/10.7705/biomedica.v32i1.386

  8. Assembly of presynaptic filaments. Factors affecting the assembly of RecA protein onto single-stranded DNA

    DEFF Research Database (Denmark)

    Thresher, RJ; Christiansen, Gunna; Griffith, JD

    1988-01-01

    We have previously shown that the assembly of RecA protein onto single-stranded DNA (ssDNA) facilitated by SSB protein occurs in three steps: (1) rapid binding of SSB protein to the ssDNA; (2) nucleation of RecA protein onto this template; and (3) co-operative polymerization of additional Rec......M in the presence of 12 mM-Mg2+), and relatively low concentrations of SSB protein (1 monomer per 18 nucleotides). Assembly was depressed threefold when SSB protein was added to one monomer per nine nucleotides. These effects appeared to be exerted at the nucleation step. Following nucleation, RecA protein...... assembled onto ssDNA at net rates that varied from 250 to 900 RecA protein monomers per minute, with the rate inversely related to the concentration of SSB protein. Combined sucrose sedimentation and electron microscope analysis established that SSB protein was displaced from the ssDNA during RecA protein...

  9. The RecX protein interacts with the RecA protein and modulates its activity in Herbaspirillum seropedicae

    Energy Technology Data Exchange (ETDEWEB)

    Galvão, C.W. [Departamento de Biologia Estrutural, Molecular e Genética, Universidade Estadual de Ponta Grossa, Ponta Grossa, PR (Brazil); Souza, E.M. [Departamento de Bioquímica e Biologia Molecular, Universidade Federal do Paraná, Curitiba, PR (Brazil); Etto, R.M. [Departamento de Biologia Estrutural, Molecular e Genética, Universidade Estadual de Ponta Grossa, Ponta Grossa, PR (Brazil); Pedrosa, F.O.; Chubatsu, L.S.; Yates, M.G. [Departamento de Bioquímica e Biologia Molecular, Universidade Federal do Paraná, Curitiba, PR (Brazil); Schumacher, J.; Buck, M. [Department of Life Sciences, Imperial College London, London (United Kingdom); Steffens, M.B.R. [Departamento de Bioquímica e Biologia Molecular, Universidade Federal do Paraná, Curitiba, PR (Brazil)

    2012-10-15

    DNA repair is crucial to the survival of all organisms. The bacterial RecA protein is a central component in the SOS response and in recombinational and SOS DNA repairs. The RecX protein has been characterized as a negative modulator of RecA activity in many bacteria. The recA and recX genes of Herbaspirillum seropedicae constitute a single operon, and evidence suggests that RecX participates in SOS repair. In the present study, we show that the H. seropedicae RecX protein (RecX{sub Hs}) can interact with the H. seropedicae RecA protein (RecA{sub Hs}) and that RecA{sub Hs} possesses ATP binding, ATP hydrolyzing and DNA strand exchange activities. RecX{sub Hs} inhibited 90% of the RecA{sub Hs} DNA strand exchange activity even when present in a 50-fold lower molar concentration than RecA{sub Hs}. RecA{sub Hs} ATP binding was not affected by the addition of RecX, but the ATPase activity was reduced. When RecX{sub Hs} was present before the formation of RecA filaments (RecA-ssDNA), inhibition of ATPase activity was substantially reduced and excess ssDNA also partially suppressed this inhibition. The results suggest that the RecX{sub Hs} protein negatively modulates the RecA{sub Hs} activities by protein-protein interactions and also by DNA-protein interactions.

  10. Development and Characterization of recA Mutants of Campylobacter jejuni for Inclusion in Attenuated Vaccines

    Science.gov (United States)

    1994-02-01

    14410 S ..-.. GII 1/I/lIIIIIHIK’II VOL. 62. 1994 C. .EJ.L!- rec.4- GENE 427 MATERIALS AND METHODS sterile broth via a nasogastric tube after...wild type. UV kill curves on VC83 and the pUCI8 probe indicated that no vector sequences were VC83 recA confirmed that the mutant was more sensitive to...vated on Mueller-Hinton (MH) agar supplemented when h of incubation, cells were suspended in brucella broth to a indicated with kanamycin (50 p.i/ml

  11. Investigating structural changes induced by nucleotide binding to RecA using difference FTIR.

    OpenAIRE

    Butler, Blaine C; Hanchett, Ross H; Rafailov, Helena; MacDonald, Gina

    2002-01-01

    Nucleotide binding to RecA results in either the high-DNA affinity form (Adenosine 5'-triphosphate (ATP)-bound) or the more inactive protein conformation associated with a lower affinity for DNA (Adenosine 5'-diphosphate (ADP)-bound). Many of the key structural differences between the RecA-ATP and RecA-ADP bound forms have yet to be elucidated. We have used caged-nucleotides and difference FTIR in efforts to obtain a comprehensive understanding of the molecular changes induced by nucleotide b...

  12. Mycobacterium tuberculosis UvrD1 and UvrA proteins suppress DNA strand exchange promoted by cognate and noncognate RecA proteins.

    Science.gov (United States)

    Singh, Pawan; Patil, K Neelakanteshwar; Khanduja, Jasbeer Singh; Kumar, P Sanjay; Williams, Alan; Rossi, Franca; Rizzi, Menico; Davis, Elaine O; Muniyappa, K

    2010-06-15

    DNA helicases are present in all kingdoms of life and play crucial roles in processes of DNA metabolism such as replication, repair, recombination, and transcription. To date, however, the role of DNA helicases during homologous recombination in mycobacteria remains unknown. In this study, we show that Mycobacterium tuberculosis UvrD1 more efficiently inhibited the strand exchange promoted by its cognate RecA, compared to noncognate Mycobacterium smegmatis or Escherichia coli RecA proteins. The M. tuberculosis UvrD1(Q276R) mutant lacking the helicase and ATPase activities was able to block strand exchange promoted by mycobacterial RecA proteins but not of E. coli RecA. We observed that M. tuberculosis UvrA by itself has no discernible effect on strand exchange promoted by E. coli RecA but impedes the reaction catalyzed by the mycobacterial RecA proteins. Our data also show that M. tuberculosis UvrA and UvrD1 can act together to inhibit strand exchange promoted by mycobacterial RecA proteins. Taken together, these findings raise the possibility that UvrD1 and UvrA might act together in vivo to counter the deleterious effects of RecA nucleoprotein filaments and/or facilitate the dissolution of recombination intermediates. Finally, we provide direct experimental evidence for a physical interaction between M. tuberculosis UvrD1 and RecA on one hand and RecA and UvrA on the other hand. These observations are consistent with a molecular mechanism, whereby M. tuberculosis UvrA and UvrD1, acting together, block DNA strand exchange promoted by cognate and noncognate RecA proteins.

  13. Direct ATP photolabeling of Escherichia coli recA proteins: identification of regions required for ATP binding

    International Nuclear Information System (INIS)

    Banks, G.R.; Sedgwick, S.G.

    1986-01-01

    When the Escherichia coli RecA protein is UV irradiated in the presence of [alpha- 32 P]ATP, a labeled protein--ATP adduct is formed. All the experimental evidence indicates that, in forming such an adduct, the ATP becomes specifically immobilized in the catalytically relevant ATP binding site. The adduct can also be identified after irradiation of E. coli cell lysates in a similar manner. This direct ATP photolabeling of RecA proteins has been used to identify regions of the polypeptide chain involved in the binding of ATP. The photolabeling of a RecA protein that lacks wild-type carboxy-terminal amino acids is not detectable. A RecA protein in which the amino-terminal sequence NH2-Ala-Ile-Asp-Glu-Asn- is replaced by NH2-Thr-Met-Ile-Thr-Asn-Ser-Ser-Ser- is only about 5% as efficiently photolabeled as the wild-type protein. Both of these RecA protein constructions, however, contain all the elements previously implicated, directly or indirectly, in the binding of ATP. ATP-photolabeled RecA protein has also been chemically cleaved at specific amino acids in order to identify regions of the polypeptide chain to which the nucleotide becomes covalently photolinked. The evidence is consistent with a region comprising amino acids 116-170. Thus, this work and that of others suggest that several disparate regions of the unfolded polypeptide chain may combine to form the ATP binding site upon protein folding or may influence binding through long-range effects

  14. Leprosy and the testis La lepra y el testículo

    Directory of Open Access Journals (Sweden)

    Fernando López

    2011-09-01

    Full Text Available Introduction. Damage of testicles is frequent in lepromatous leprosy worsen by the presence of erythema nodosum leprosum.
    Objective. One patient is presented who developed lepromatous leprosy and erythema nodosum leprosum with important testicular compromise.
    Material and methods. A 28 year old man patient who had lepromatous leprosy since his 22 was studied. During a polychemotherapy treatment for the lepromatous leprosy, he presented chronic erythema nodosum leprosum that affected both testicles and did not respond to the conventional treatment. A left orchidectomy was practiced to treat the persistent pain.
    Results. The extracted testis evidenced the following findings: tubular atrophy, remarkable fibrosis, cumulus of foamy macrophages without rods, focal Leydig cell hyperplasia, linfocitary and granulomatous arteritis and endarteritis of small and medium size vessels. These changes were also observed in the epididymis. Two years after the polychemoterapy and the orchidectomy, the patient revealed azoospermy, normal total testosterone, discretely diminished free testosterone and elevated luteinizing hormone and follicle-stimulating hormone. No loss of libido or sexual activity. We reviewed general concepts about erythema nodosum leprosum and the pathologic changes produced by leprosy in the testis.
    Conclusion. Lepromatous leprosy may lead to hypogonadism. This should be considered by the leprosy programs in order to avoid and treat the consequences of the possible hypogonadism.
    Introducción. La afección testicular es frecuente en la lepra lepromatosa, daño que se incrementa cuando cursa con eritema nodoso leproso.
    Objetivo. Presentar un paciente con lepra lepromatosa y eritema nodoso leproso con severo compromiso testicular.
    Materiales y métodos. Estudiamos un hombre de 28 años con lepra lepromatosa desde los 22, que durante el tratamiento con poliquimioterapia para la lepra presentó eritema nodoso

  15. Serological monitoring of previously treated lepromatous patients during a course of multiple immunotherapy treatments with heat-killed Mycobacterium leprae and BCG

    NARCIS (Netherlands)

    Douglas, J. T.; Hirsch, D. S.; Fajardo, T. T.; Guido, L. S.; Klatser, P. R.

    1990-01-01

    Two-hundred and seventy lepromatous patients who had completed treatment received multiple treatments with heat-killed M. leprae and BCG and were monitored for changes in humoral responses to M. leprae-specific antigens. These patients were divided into four treatment groups: placebo (n = 69); BCG

  16. Correlation of bacterial viability with uptake of (14C) acetate into phenolic glycolipid-1 of Mycobacterium leprae within Schwannoma cells

    International Nuclear Information System (INIS)

    Mistry, Y.; Antia, N.H.; Mukherjee, R.

    1989-01-01

    The viability of Mycobacterium leprae, maintained within 33B Schwannoma cells, was estimated in terms of incorporation of ( 14 C) acetate into its specific phenolic glycolipid-1. This measure of viability was correlated with two other assays, viz., fluorescein diacetate/ethidium bromide staining and mouse footpad growth. Observation of a 2-fold increase in the number of intracellular Mycobacterium leprae over an experimental period of 12 days also corroborated this contention. Furthermore, on addition of anti-leprosy drugs to these intracellular Mycobacterium leprae there was significant decrease in phenolic glycolipid-1 synthesis indicative of loss of viability of the organisms. This study also established the importance of the host cell for active bacillary metabolism, as Mycobacterium leprae maintained in cell-free conditions showed no incorporation into phenolic glycolipid-1. Moreover, compromising the host's protein synthesis capacity with cycloheximide, also led to reduction in bacillary metabolism. As this system measures the metabolic synthesis of a unique Mycobacterium leprae component, it would be useful for development and screening of compounds acting against specific bacillary targets. (author). 19 refs., 5 tabs

  17. Mycobacterium leprae in six-banded (Euphractus sexcinctus and nine-banded armadillos (Dasypus novemcinctus in Northeast Brazil

    Directory of Open Access Journals (Sweden)

    Cristiane Cunha Frota

    2012-12-01

    Full Text Available Human beings are the main reservoir of the causative agent of leprosy, Mycobacterium leprae. In the Americas, nine-banded armadillos (Dasypus novemcinctus also act as a reservoir for the bacillus. In the state of Ceará (CE, which is located in Northeast Brazil and is an endemic area of leprosy, there are several species of armadillos, including D. novemcinctus and Euphractus sexcinctus (six-banded armadillo. Contact between humans and armadillos occur mainly through hunting, cleaning, preparing, cooking and eating. This study identified M. leprae DNA in the two main species of armadillos found in Northeast Brazil. A total of 29 wild armadillos (27 D. novemcinctus and 2 E. sexcinctus were captured in different environments of CE countryside. Samples from the ear, nose, liver and spleen from each of these animals were tested by a nested M. leprae-specific repetitive element polymerase chain reaction assay. The samples that tested positive were confirmed by DNA sequencing. M. leprae was detected in 21% (6/29 of the animals, including five D. novemcinctus and one E. sexcinctus. This is the first Brazilian study to identify the presence of a biomarker of M. leprae in wild armadillos (D. novemcinctus and E. sexcinctus in a leprosy hyperendemic area where there is continuous contact between humans and armadillos.

  18. Nasal PCR assay for the detection of Mycobacterium leprae pra gene to study subclinical infection in a community.

    Science.gov (United States)

    Arunagiri, Kamalanathan; Sangeetha, Gopalakrishnan; Sugashini, Padmavathy Krishnan; Balaraman, Sekar; Showkath Ali, M K

    2017-03-01

    Leprosy is a chronic infectious disease caused by Mycobacterium leprae. Identification of Mycobacterium leprae is difficult in part due to the inability of the leprosy bacillus to grow in vitro. A number of diagnostic methods for leprosy diagnosis have been proposed. Both serological tests and molecular probes have shown certain potential for detection and identification of Mycobacterium leprae in patients. In this study, we have investigated whether Mycobacterium leprae DNA from the nasal secretion of healthy household contacts and the non contacts could be detected through PCR amplification as a method to study the sub clinical infection in a community. A total of 200 samples, 100 each from contacts and non contacts representing all age groups and sex were included in this study. The M. leprae specific primer (proline-rich region) of pra gene was selected and PCR was performed using extracted DNA from the sample. A total of 13 samples were found to be positive for nasal PCR for pra gene among the male and female contacts out of which 7% were males and 6% were females. Even though several diagnostic tools are available to detect the cases of leprosy, they lack the specificity and sensitivity. PCR technology has demonstrated the improved diagnostic accuracy for epidemiological studies and requires minimal time. Although nasal PCR studies have been reported from many countries it is not usually recommended due to the high percentage of negative results in the contact. Copyright © 2017 Elsevier Ltd. All rights reserved.

  19. Genetic control of near-UV (300-400 nm) sensitivity independent of the recA gene in strains of Escherichia coli K12

    International Nuclear Information System (INIS)

    Tuveson, R.W.; Jonas, R.B.

    1979-01-01

    Stationary cells of isogenic pairs of Escherichia coli K12 strains presumably differing only in the recA function, were inactivated with near-UV (300-400 nm) radiation. Based on near-UV inactivation kinetics, the strains can be divided into two discrete categories in which near-UV sensitivity does not necessarily correlate with far-UV sensitivity conferred by two different recA alleles. Lack of overlap between near-UV and far-UV (recA) sensitivity can be explained by assuming that a different chromosomal gene (nur) controls near-UV sensitivity. Support for this hypothesis came from a mating experiment in which four selected recombinants, isogenic with respect to auxotrophic markers, were identified exhibiting all four possible combinations of far-UV (recA1 vs recA + ) and near-UV sensitivity (nur vs nur + ). Transduction with phase P1 showed that introduction of the recA1 allele into a recA + recipient did not affect the near-UV sensitivity of the recipient. Additional matings together with transduction experiments suggested that the nur gene is located at a position on the E. coli linkage map clearly separable from recA (minute 58). (author)

  20. In vitro incorporation of a cell-binding protein to a lentiviral vector using an engineered split intein enables targeted delivery of genetic cargo.

    Science.gov (United States)

    Chamoun-Emanuelli, Ana M; Wright, Gus; Roger, Smith; Münch, Robert C; Buchholz, Christian J; Chen, Zhilei

    2015-12-01

    Gene therapy represents a promising therapeutic paradigm for addressing many disorders, but the absence of a vector that can be robustly and reproducibly functionalized with cell-homing functionality to mediate the delivery of genetic cargo specifically to target cells following systemic administration has stood as a major impediment. In this study, a high-affinity protein-protein pair comprising a splicing-deficient naturally split intein was used as molecular Velcro to append a HER2/neu-binding protein (DARPin) onto the surface of a binding-deficient, fusion-competent lentivirus. HER2/neu-specific lentiviruses created using this in vitro pseudotyping approach were able to deliver their genetic reporter cargo specifically to cells that express the target receptor at high levels in a co-culture. We envision that the described technology could provide a powerful, broadly applicable platform for the incorporation of cell-targeting functionality onto viral vectors. © 2015 Wiley Periodicals, Inc.

  1. Métodos tintoriais utilizados na identificação do Mycobacterium leprae: revisão histológica Staining methods used in the identification of Mycobacterium leprae: historical review

    Directory of Open Access Journals (Sweden)

    Luiz Fernando de Góes Siqueira

    1984-06-01

    Full Text Available Foi feita revisão histórica sobre métodos tintoriais utilizados na identificação baciloscópica do Mycobacterium leprae. Ao lado da descrição de cada método, e suas variantes, é feita extensa revisão bibliográfica.A historical review of the staining methods utilized in the bacilloscopic identification of the Mycobacterium leprae was made. Beside the description of each method and its variants, an extensive bibliographical review is made.

  2. Dickeya species relatedness and clade structure determined by comparison of recA sequences.

    Science.gov (United States)

    Parkinson, Neil; Stead, David; Bew, Janice; Heeney, John; Tsror Lahkim, Leah; Elphinstone, John

    2009-10-01

    Using sequences from the recA locus, we have produced a phylogeny of 188 Dickeya strains from culture collections and identified species relatedness and subspecies clade structure within the genus. Of the six recognized species, Dickeya paradisiaca, D. chrysanthemi and D. zeae were discriminated with long branch lengths. The clade containing the D. paradisiaca type strain included just one additional strain, isolated from banana in Colombia. Strains isolated from Chrysanthemum and Parthenium species made up most of the clade containing the D. chrysanthemi type strain, and the host range of this species was extended to include potato. The D. zeae clade had the largest number of sequevars and branched into two major sister clades that contained all of the Zea mays isolates, and were identified as phylotypes PI and PII. The host range was increased from six to 13 species, including potato. The recA sequence of an Australian sugar-cane strain was sufficiently distinct to rank as a new species-level branch. In contrast to these species, Dickeya dadantii, D. dianthicola and D. dieffenbachiae were distinguished with shorter branch lengths, indicating relatively closer relatedness. The recA sequence for the type strain of D. dadantii clustered separately from other strains of the species. However, sequence comparison of three additional loci revealed that the D. dadantii type strain grouped together with the six other D. dadantii strains that were sequenced. Analysis of all four loci indicated that the D. dadantii strains were most closely related to D. dieffenbachiae. Three further branches (DUC-1, -2 and -3) were associated with these three species, which all diverged from a common origin and can be considered as a species complex. The large clade containing the D. dianthicola type strain comprised 58 strains and had little sequence diversity. One sequevar accounted for the majority of these strains, which were isolated nearly exclusively from eight hosts from Europe

  3. Expression of the recA gene of Pseudomonas aeruginosa PAO is inducible by DNA-damaging agents

    International Nuclear Information System (INIS)

    Miller, R.V.; Kokjohn, T.A.

    1988-01-01

    Western (immunoblot) analysis using Escherichia coli anti-RecA antiserum revealed that expression of the RecA protein of Pseudomonas aeruginosa PAO is induced upon exposure of the bacterium to UV irradiation or norfloxacin, a quinolone related to nalidixic acid

  4. Role of RecA and the SOS response in thymineless death in Escherichia coli.

    Directory of Open Access Journals (Sweden)

    Natalie C Fonville

    2010-03-01

    Full Text Available Thymineless death (TLD is a classic and enigmatic phenomenon, documented in bacterial, yeast, and human cells, whereby cells lose viability rapidly when deprived of thymine. Despite its being the essential mode of action of important chemotherapeutic agents, and despite having been studied extensively for decades, the basic mechanisms of TLD have remained elusive. In Escherichia coli, several proteins involved in homologous recombination (HR are required for TLD, however, surprisingly, RecA, the central HR protein and activator of the SOS DNA-damage response was reported not to be. We demonstrate that RecA and the SOS response are required for a substantial fraction of TLD. We show that some of the Rec proteins implicated previously promote TLD via facilitating activation of the SOS response and that, of the roughly 40 proteins upregulated by SOS, SulA, an SOS-inducible inhibitor of cell division, accounts for most or all of how SOS causes TLD. The data imply that much of TLD results from an irreversible cell-cycle checkpoint due to blocked cell division. FISH analyses of the DNA in cells undergoing TLD reveal blocked replication and apparent DNA loss with the region near the replication origin underrepresented initially and the region near the terminus lost later. Models implicating formation of single-strand DNA at blocked replication forks, a SulA-blocked cell cycle, and RecQ/RecJ-catalyzed DNA degradation and HR are discussed. The data predict the importance of DNA damage-response and HR networks to TLD and chemotherapy resistance in humans.

  5. High frequency of M. leprae DNA detection in asymptomatic household contacts.

    Science.gov (United States)

    Gama, Rafael Silva; Gomides, Thalisson Artur Ribeiro; Gama, Chaiana Fróes Magalhães; Moreira, Suelen Justo Maria; de Neves Manta, Fernanda Saloum; de Oliveira, Lorena Bruna P; Marçal, Pedro Henrique Ferreira; Sarno, Euzenir Nunes; Moraes, Milton Ozório; Garcia, Raúl Marcel González; de Oliveira Fraga, Lucia Alves

    2018-04-02

    Characterization of the Mycobacterium leprae genome has made possible the development of Polymerase Chain Reaction (PCR) systems that can amplify different genomic regions. Increased reliability and technical efficiency of quantitative PCR (qPCR) makes it a promising tool for early diagnosis of leprosy. Index cases that are multibacillary spread the bacillus silently, even before they are clinically diagnosed. Early detection and treatment could prevent transmission in endemic areas. In this study, the qPCR technique is used to detect DNA of M. leprae in samples of slit skin smears (SSS) of the ear lobe and blood of leprosy patients and their asymptomatic household contacts residing in Governador Valadares, MG, Brazil, a hyperendemic area for leprosy. A total of 164 subjects participated in the study: 43 index cases, 113 household contacts, and, as negative controls, 8 individuals who reported no contact with patients nor history of leprosy in the family. The qPCR was performed to amplify 16S rRNA fragments and was specifically designed for M. leprae. Of asymptomatic household contacts, 23.89% showed bacillary DNA by qPCR in samples of SSS and blood. Also, 48.84% of patients diagnosed with leprosy were positive for qPCR while the bacillary load was positive in only 30.23% of patients. It is important to note that most patients were already receiving treatment when the collection of biological material for qPCR was performed. The level of bacillary DNA from household contacts was similar to the DNA levels detected in the group of paucibacillary patients. Considering that household contacts comprise a recognizable group of individuals with a high risk of disease, as they live in close proximity to a source of infection, qPCR can be used to estimate the risk of progress towards leprosy among household contacts and as a routine screening method for a chemoprophylactic protocol.

  6. Comportamiento de la lepra en la provincia de Las Tunas, 2003-2012

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    Martha O León Cabrales

    2014-08-01

    Full Text Available Se realizó un estudio descriptivo de corte transversal, para determinar algunas características epidemiológicas de la incidencia de la lepra, que es una enfermedad transmisible, tan antigua como el hombre mismo. El universo estuvo constituido por los 103 casos notificados con lepra en la provincia de Las Tunas, en el período de enero de 2003 a diciembre de 2012. La información se obtuvo por las encuestas epidemiológicas existentes en el Departamento de Estadística de la Dirección Provincial de Salud y en el Centro Provincial de Higiene, Epidemiología y Microbiología. Se creó una base de datos en Epinfo versión 3.3.3, donde se tabularon los datos de las encuestas. El análisis de los resultados se expresó en números absolutos, tasas y porcentajes para su mejor interpretación, obteniéndose como resultado que la tasa de detección de casos tiene un comportamiento irregular, el año de mayor incidencia fue el 2009, con 20 casos. Se notificaron tres casos de lepra infantil; las formas paucibacilares representaron el 51,5%; el modo de detección más frecuente fue el espontáneo. Existe transmisión activa y todo ello puede ser reflejo de la ausencia de un trabajo consolidado en el programa de control de la enfermedad

  7. A novel role for RecA under non-stress: promotion of swarming motility in Escherichia coli K-12

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    Blázquez Jesús

    2007-03-01

    Full Text Available Abstract Background Bacterial motility is a crucial factor in the colonization of natural environments. Escherichia coli has two flagella-driven motility types: swimming and swarming. Swimming motility consists of individual cell movement in liquid medium or soft semisolid agar, whereas swarming is a coordinated cellular behaviour leading to a collective movement on semisolid surfaces. It is known that swimming motility can be influenced by several types of environmental stress. In nature, environmentally induced DNA damage (e.g. UV irradiation is one of the most common types of stress. One of the key proteins involved in the response to DNA damage is RecA, a multifunctional protein required for maintaining genome integrity and the generation of genetic variation. Results The ability of E. coli cells to develop swarming migration on semisolid surfaces was suppressed in the absence of RecA. However, swimming motility was not affected. The swarming defect of a ΔrecA strain was fully complemented by a plasmid-borne recA gene. Although the ΔrecA cells grown on semisolidsurfaces exhibited flagellar production, they also presented impaired individual movement as well as a fully inactive collective swarming migration. Both the comparative analysis of gene expression profiles in wild-type and ΔrecA cells grown on a semisolid surface and the motility of lexA1 [Ind-] mutant cells demonstrated that the RecA effect on swarming does not require induction of the SOS response. By using a RecA-GFP fusion protein we were able to segregate the effect of RecA on swarming from its other functions. This protein fusion failed to regulate the induction of the SOS response, the recombinational DNA repair of UV-treated cells and the genetic recombination, however, it was efficient in rescuing the swarming motility defect of the ΔrecA mutant. The RecA-GFP protein retains a residual ssDNA-dependent ATPase activity but does not perform DNA strand exchange. Conclusion

  8. Mycobacterium leprae downregulates the expression of PHEX in Schwann cells and osteoblasts

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    Sandra R Boiça Silva

    2010-08-01

    Full Text Available Neuropathy and bone deformities, lifelong sequelae of leprosy that persist after treatment, result in significant impairment to patients and compromise their social rehabilitation. Phosphate-regulating gene with homologies to endopeptidase on the X chromosome (PHEX is a Zn-metalloendopeptidase, which is abundantly expressed in osteoblasts and many other cell types, such as Schwann cells, and has been implicated in phosphate metabolism and X-linked rickets. Here, we demonstrate that Mycobacterium leprae stimulation downregulates PHEX transcription and protein expression in a human schwannoma cell line (ST88-14 and human osteoblast lineage. Modulation of PHEX expression was observed to a lesser extent in cells stimulated with other species of mycobacteria, but was not observed in cultures treated with latex beads or with the facultative intracellular bacterium Salmonella typhimurium. Direct downregulation of PHEX by M. leprae could be involved in the bone resorption observed in leprosy patients. This is the first report to describe PHEX modulation by an infectious agent.

  9. La lepra, un problema de salud global Leprosy is a problem of global health

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    Eduardo Rivero Reyes

    2009-03-01

    Full Text Available Le lepra constituye una enfermedad conocida desde el año 2000 a.n.e., que causó verdaderos estragos a poblaciones enteras y azotó ininterrumpidamente a todos los continentes. Ha sido considerada una enfermedad mutilante, incurable, repulsiva y estigmatizante, lo que ha generado un trato inhumano hacia las personas que la padecen, constituyendo aún hoy un problema de salud importante para algunos países. En el presente artículo se describen algunos apuntes sobre la evolución histórica de la lepra en Cuba y a nivel mundial.Leprosy is a known disease from 2000 b.o.e, causing real havocs in complete populations and lashed continuously to all continents. Has been considered a mutilating, incurable, repulsive and stigmatizing disease that has generated a inhuman deal towards leprous persons, even nowadays is very important health problem for some countries. In present paper some notes on historical evolution of leprosy in Cuba and at world scale are described.

  10. Fatores de risco que favorecem a recaída no alcoolismo Risk factors that favor the relapse in alcoholism

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    Armando M. Alonso Álvarez

    2007-01-01

    Full Text Available OBJETIVO: Existe uma prevalência estimada de 11,2% de alcoolismo no Brasil. As tentativas de muitos alcoolistas para abandonar o consumo resultam fracassos e recaídas. A freqüência de recaída, segundo estudos, oscila entre 10% e 30%, assinalando-se diferentes causas. O objetivo da presente pesquisa é conhecer os fatores que favorecem as recaídas. MÉTODOS: Aplicaram-se entrevistas e questionários "Fatores de Recaída" e "Razões para Beber" a 105 alcoolistas primários de ambos os sexos, e com predomínio de alcoolismo puro. RESULTADOS: Os principais fatores de recaída são a pressão social, os conflitos interpessoais, os estados emocionais negativos, assim como a própria dependência fisiológica e psicológica. CONCLUSÕES: De maneira geral, os sujeitos apresentaram grande quantidade de fatores e razões que atuam simultaneamente favorecendo a recaída.OBJETIVE: There is an estimate of 11.2% of alcohol prevalence in Brazil. The attempts of many alcoholics for abandoning the addiction result in failures and relapse. The number of relapse oscillates between 10 and 30%, designating different causes. The objective of the present research is to know the factors that favor relapses. METHODS: interviews and the questionnaires "Relapse Factors" and "Reasons for Drink" were applied, to 105 alcoholics of both sexes, with primary and typical alcoholism. RESULTS: Initially, social demographics data are presented followed by a characterization of the information gotten in the interview and each questionnaire, which are integrated and submitted in discussion. CONCLUSIONS: the main factors are: the social pressure, the interpersonal conflicts, the negative emotional states as well as the proper physiological and psychological dependence. In general way, they had presented a great amount of factors and reasons that simultaneously act favoring the relapse.

  11. Unique TTC repeat base pair loss mutation in cases of pure neural leprosy: A survival strategy of Mycobacterium leprae?

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    Abhishek De

    2015-01-01

    Full Text Available Background: Genomic reduction helps obligate intracellular microbes to survive difficult host niches. Adaptation of Mycobacterium leprae in cases of pure neural leprosy (PNL in the intracellular niche of peripheral nerves can be associated with some gene loss. Recently, a stable but variable number of tandem repefzats (TTC have been reported in strains of M. leprae. FolP and rpoB genes are the two common mutation sites which deal with the susceptibility of the bacteria to drugs. Aim: We attempted to find if genomic reduction of M. leprae in context of these TTC repeats or mutations in folP1 and rpoB can be the reason for the restriction of M. leprae in the nerves in PNL. Materials and Methods: DNA extracts taken from fine needle aspiration of affected nerves of 24 PNL cases were studied for tandem repeats with 21TTC primer in multiplex-PCR. Mutations were also studied by PCR Amplification of SRDR (Sulphone Resistance Determining Region of the folP1 and multiple primer PCR amplification refractory mutation system (MARS of the rpoB. Results: Of the 24 PNL, only 1 patient showed mutation in the rpoB gene and none in the folp1 gene. Studying the mutation in TTC region of the M. leprae gene we found that all the cases have a loss of a few bases in the sequence. Conclusion: We can conclude that there is consistent loss in the bases in the TTC region in all cases of pure neural Hansen and we postulate that it may be an adaptive response of the bacteria to survive host niche resulting in its restriction to peripheral nerves.

  12. Human Genetic Ancestral Composition Correlates with the Origin of Mycobacterium leprae Strains in a Leprosy Endemic Population.

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    Nora Cardona-Castro

    Full Text Available Recent reports have suggested that leprosy originated in Africa, extended to Asia and Europe, and arrived in the Americas during European colonization and the African slave trade. Due to colonization, the contemporary Colombian population is an admixture of Native-American, European and African ancestries. Because microorganisms are known to accompany humans during migrations, patterns of human migration can be traced by examining genomic changes in associated microbes. The current study analyzed 118 leprosy cases and 116 unrelated controls from two Colombian regions endemic for leprosy (Atlantic and Andean in order to determine possible associations of leprosy with patient ancestral background (determined using 36 ancestry informative markers, Mycobacterium leprae genotype and/or patient geographical origin. We found significant differences between ancestral genetic composition. European components were predominant in Andean populations. In contrast, African components were higher in the Atlantic region. M. leprae genotypes were then analyzed for cluster associations and compared with the ancestral composition of leprosy patients. Two M. leprae principal clusters were found: haplotypes C54 and T45. Haplotype C54 associated with African origin and was more frequent in patients from the Atlantic region with a high African component. In contrast, haplotype T45 associated with European origin and was more frequent in Andean patients with a higher European component. These results suggest that the human and M. leprae genomes have co-existed since the African and European origins of the disease, with leprosy ultimately arriving in Colombia during colonization. Distinct M. leprae strains followed European and African settlement in the country and can be detected in contemporary Colombian populations.

  13. Overexpression of Salmonella enterica serovar Typhi recA gene confers fluoroquinolone resistance in Escherichia coli DH5α

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    M.A.M. Yassien

    2015-11-01

    Full Text Available A spontaneous fluoroquinolone-resistant mutant (STM1 was isolated from its parent Salmonella enterica serovar Typhi (S. Typhi clinical isolate. Unlike its parent isolate, this mutant has selective resistance to fluoroquinolones without any change in its sensitivity to various other antibiotics. DNA gyrase assays revealed that the fluoroquinolone resistance phenotype of the STM1 mutant did not result from alteration of the fluoroquinolone sensitivity of the DNA gyrase isolated from it. To study the mechanism of fluoroquinolone resistance, a genomic library from the STM1 mutant was constructed in Escherichia coli DH5α and two recombinant plasmids were obtained. Only one of these plasmids (STM1-A conferred the selective fluoroquinolone resistance phenotype to E. coli DH5α. The chromosomal insert from STM1-A, digested with EcoRI and HindIII restriction endonucleases, produced two DNA fragments and these were cloned separately into pUC19 thereby generating two new plasmids, STM1-A1 and STM1-A2. Only STM1-A1 conferred the selective fluoroquinolone resistance phenotype to E. coli DH5α. Sequence and subcloning analyses of STM1-A1 showed the presence of an intact RecA open reading frame. Unlike that of the wild-type E. coli DH5α, protein analysis of a crude STM1-A1 extract showed overexpression of a 40 kDa protein. Western blotting confirmed the 40 kDa protein band to be RecA. When a RecA PCR product was cloned into pGEM-T and introduced into E. coli DH5α, the STM1-A11 subclone retained fluoroquinolone resistance. These results suggest that overexpression of RecA causes selective fluoroquinolone resistance in E. coli DH5α.

  14. recA and catalase in H sub 2 O sub 2 -mediated toxicity in Neisseria gonorrhoeae

    Energy Technology Data Exchange (ETDEWEB)

    Hassett, D.J.; Charniga, L.; Cohen, M.S. (Univ. of North Carolina, Chapel Hill (USA))

    1990-12-01

    Neisseria gonorrhoeae cells defective in the biosynthesis of the recA gene product are no more sensitive to hydrogen peroxide than wild-type cells. Although gonococci possess nearly 100-fold-greater catalase levels than Escherichia coli, they are more susceptible to hydrogen peroxide than this organism. The natural niche of gonococci undoubtedly results in exposure to oxidant stress; however, they do not demonstrate particularly efficient antioxidant defense systems.

  15. Factores psicosociales en la recaída de la dependencia al alcohol: Un análisis de ruta

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    Fabiola Alejandra Useche Torrealba

    2017-12-01

    Full Text Available Introducción: El consumo de alcohol causa alrededor del 6% de muertes a nivel mundial y se espera que el 70% de los pacientes en recuperación recaigan en los primeros seis meses de tratamiento. Esto, aunado a la aceptación social de esta sustancia y su fácil acceso, hacen necesario estudiar las variables asociadas a la recaída. Objetivo: Estudiar la influencia de variables sociodemográficas y psicosociales, sobre la recaída de la dependencia al alcohol. Métodos: Se realizó una investigación de campo, transversal y explicativa-correlacional, con la participación de 180 miembros de Alcohólicos Anónimos de Venezuela. Resultados: Participaron 87 mujeres y 93 hombres, con un promedio de 50 años. El índice de recaída fue de 18%, con un promedio de 130 meses de abstinencia. Se encontraron altos niveles de apoyo social, autoeficacia y autoestima, así como valores intermedios de estrés, impulsividad y resiliencia. Las variables estudiadas se asociaron con el tiempo en abstinencia (R = 0.615; p < 0.05 explicando el 35% de la varianza total. La edad (β = 0.57; p = 0.00, autoestima (β = -0.156; p = 0.02, apoyo social (β = 0.148; p = 0.02 y sexo (β = -0.135; p = 0.03 fueron las variables con mayor eficacia en la ruta principal. Conclusiones: El modelo propuesto se cumplió parcialmente. Es necesario profundizar el estudio de relaciones no planteadas entre las variables moderadoras, y si estas tienen otras que las modifiquen. Se recomienda enfatizar el abordaje de las variables que resultaron relevantes en la prevención de la recaída.

  16. Overexpression of Salmonella enterica serovar Typhi recA gene confers fluoroquinolone resistance in Escherichia coli DH5α.

    Science.gov (United States)

    Yassien, M A M; Elfaky, M A

    2015-11-01

    A spontaneous fluoroquinolone-resistant mutant (STM1) was isolated from its parent Salmonella enterica serovar Typhi (S. Typhi) clinical isolate. Unlike its parent isolate, this mutant has selective resistance to fluoroquinolones without any change in its sensitivity to various other antibiotics. DNA gyrase assays revealed that the fluoroquinolone resistance phenotype of the STM1 mutant did not result from alteration of the fluoroquinolone sensitivity of the DNA gyrase isolated from it. To study the mechanism of fluoroquinolone resistance, a genomic library from the STM1 mutant was constructed in Escherichia coli DH5α and two recombinant plasmids were obtained. Only one of these plasmids (STM1-A) conferred the selective fluoroquinolone resistance phenotype to E. coli DH5α. The chromosomal insert from STM1-A, digested with EcoRI and HindIII restriction endonucleases, produced two DNA fragments and these were cloned separately into pUC19 thereby generating two new plasmids, STM1-A1 and STM1-A2. Only STM1-A1 conferred the selective fluoroquinolone resistance phenotype to E. coli DH5α. Sequence and subcloning analyses of STM1-A1 showed the presence of an intact RecA open reading frame. Unlike that of the wild-type E. coli DH5α, protein analysis of a crude STM1-A1 extract showed overexpression of a 40 kDa protein. Western blotting confirmed the 40 kDa protein band to be RecA. When a RecA PCR product was cloned into pGEM-T and introduced into E. coli DH5α, the STM1-A11 subclone retained fluoroquinolone resistance. These results suggest that overexpression of RecA causes selective fluoroquinolone resistance in E. coli DH5α.

  17. Evaluation of Mycobacterium leprae antigens in the serological monitoring of a clofazimine-based chemotherapeutic study of dapsone resistant lepromatous leprosy patients in Cebu, Philippines

    NARCIS (Netherlands)

    Douglas, J. T.; Hirsch, D. S.; Fajardo, T. T.; Cellona, R. V.; Abalos, R. M.; de la Cruz, E. C.; Madarang, M. G.; de Wit, M. Y.; Klatser, P. R.

    1989-01-01

    Thirty-one dapsone resistant lepromatous leprosy patients receiving clofazimine based therapy were serologically monitored throughout their 5-year period of treatment. Sequentially collected sera were used to examine 4 Mycobacterium leprae antigens to evaluate their usefulness in ELISA's for

  18. Correlated motion of protein subdomains and large-scale conformational flexibility of RecA protein filament

    Science.gov (United States)

    Yu, Garmay; A, Shvetsov; D, Karelov; D, Lebedev; A, Radulescu; M, Petukhov; V, Isaev-Ivanov

    2012-02-01

    Based on X-ray crystallographic data available at Protein Data Bank, we have built molecular dynamics (MD) models of homologous recombinases RecA from E. coli and D. radiodurans. Functional form of RecA enzyme, which is known to be a long helical filament, was approximated by a trimer, simulated in periodic water box. The MD trajectories were analyzed in terms of large-scale conformational motions that could be detectable by neutron and X-ray scattering techniques. The analysis revealed that large-scale RecA monomer dynamics can be described in terms of relative motions of 7 subdomains. Motion of C-terminal domain was the major contributor to the overall dynamics of protein. Principal component analysis (PCA) of the MD trajectories in the atom coordinate space showed that rotation of C-domain is correlated with the conformational changes in the central domain and N-terminal domain, that forms the monomer-monomer interface. Thus, even though C-terminal domain is relatively far from the interface, its orientation is correlated with large-scale filament conformation. PCA of the trajectories in the main chain dihedral angle coordinate space implicates a co-existence of a several different large-scale conformations of the modeled trimer. In order to clarify the relationship of independent domain orientation with large-scale filament conformation, we have performed analysis of independent domain motion and its implications on the filament geometry.

  19. Characterization of the Pseudomonas aeruginosa recA gene: the Les- phenotype

    International Nuclear Information System (INIS)

    Kokjohn, T.A.; Miller, R.V.

    1988-01-01

    The Les- phenotype (lysogeny establishment deficient) is a pleiotropic effect of the lesB908 mutation of Pseudomonas aeruginosa PAO. lesB908-containing strains are also (i) deficient in general recombination, (ii) sensitive to UV irradiation, and (iii) deficient in UV-stimulated induction of prophages. The P. aeruginosa recA-containing plasmid pKML3001 complemented each of these pleiotropic characteristics of the lesB908 mutation, supporting the hypothesis that lesB908 is an allele of the P. aeruginosa recA gene. The phenotypic effects of the lesB908 mutation may be best explained by the hypothesis that the lesB908 gene product is altered in such a way that it has lost synaptase activity but possesses intrinsic protease activity in the absence of DNA damage. The Les- phenotype is a result of the rapid destruction of newly synthesized phage repressor, resulting in lytic growth of the infecting virus. This hypothesis is consistent with the observations that increasing the number of copies of the phage repressor gene by increasing the multiplicity of infection (i.e., average number of phage genomes per cell) or by introducing the cloned phage repressor gene into a lesB908 mutant will also suppress the Les- phenotype in a phage-specific fashion

  20. DNA fingerprinting of Mycobacterium leprae strains using variable number tandem repeat (VNTR) - fragment length analysis (FLA).

    Science.gov (United States)

    Jensen, Ronald W; Rivest, Jason; Li, Wei; Vissa, Varalakshmi

    2011-07-15

    The study of the transmission of leprosy is particularly difficult since the causative agent, Mycobacterium leprae, cannot be cultured in the laboratory. The only sources of the bacteria are leprosy patients, and experimentally infected armadillos and nude mice. Thus, many of the methods used in modern epidemiology are not available for the study of leprosy. Despite an extensive global drug treatment program for leprosy implemented by the WHO, leprosy remains endemic in many countries with approximately 250,000 new cases each year. The entire M. leprae genome has been mapped and many loci have been identified that have repeated segments of 2 or more base pairs (called micro- and minisatellites). Clinical strains of M. leprae may vary in the number of tandem repeated segments (short tandem repeats, STR) at many of these loci. Variable number tandem repeat (VNTR) analysis has been used to distinguish different strains of the leprosy bacilli. Some of the loci appear to be more stable than others, showing less variation in repeat numbers, while others seem to change more rapidly, sometimes in the same patient. While the variability of certain VNTRs has brought up questions regarding their suitability for strain typing, the emerging data suggest that analyzing multiple loci, which are diverse in their stability, can be used as a valuable epidemiological tool. Multiple locus VNTR analysis (MLVA) has been used to study leprosy evolution and transmission in several countries including China, Malawi, the Philippines, and Brazil. MLVA involves multiple steps. First, bacterial DNA is extracted along with host tissue DNA from clinical biopsies or slit skin smears (SSS). The desired loci are then amplified from the extracted DNA via polymerase chain reaction (PCR). Fluorescently-labeled primers for 4-5 different loci are used per reaction, with 18 loci being amplified in a total of four reactions. The PCR products may be subjected to agarose gel electrophoresis to verify the

  1. Intracellular d-Serine Accumulation Promotes Genetic Diversity via Modulated Induction of RecA in Enterohemorrhagic Escherichia coli.

    Science.gov (United States)

    Connolly, James P R; Roe, Andrew J

    2016-12-15

    We recently discovered that exposure of enterohemorrhagic Escherichia coli (EHEC) to d-serine resulted in accumulation of this unusual amino acid, induction of the SOS regulon, and downregulation of the type III secretion system that is essential for efficient colonization of the host. Here, we have investigated the physiological relevance of this elevated SOS response, which is of particular interest given the presence of Stx toxin-carrying lysogenic prophages on the EHEC chromosome that are activated during the SOS response. We found that RecA elevation in response to d-serine, while being significant, was heterogeneous and not capable of activating stx expression or stx phage transduction to a nonlysogenic recipient. This "SOS-like response" was, however, capable of increasing the mutation frequency associated with low-level RecA activity, thus promoting genetic diversity. Furthermore, this response was entirely dependent on RecA and enhanced in the presence of a DNA-damaging agent, indicating a functional SOS response, but did not result in observable cleavage of the LexA repressor alone, indicating a controlled mechanism of induction. This work demonstrates that environmental factors not usually associated with DNA damage are capable of promoting an SOS-like response. We propose that this modulated induction of RecA allows EHEC to adapt to environmental insults such as d-serine while avoiding unwanted phage-induced lysis. The SOS response is a global stress network that is triggered by the presence of DNA damage due to breakage or stalled replication forks. Activation of the SOS response can trigger the replication of lytic bacteriophages and promote genetic diversification through error-prone polymerases. We have demonstrated that the host-associated metabolite d-serine contributes to Escherichia coli niche specification and accumulates inside cells that cannot catabolize it. This results in a modulated activation of the SOS antirepressor RecA that is

  2. Mycobacterium leprae in Colombia described by SNP7614 in gyrA, two minisatellites and geography

    Science.gov (United States)

    Cardona-Castro, Nora; Beltrán-Alzate, Juan Camilo; Romero-Montoya, Irma Marcela; Li, Wei; Brennan, Patrick J; Vissa, Varalakshmi

    2013-01-01

    New cases of leprosy are still being detected in Colombia after the country declared achievement of the WHO defined ‘elimination’ status. To study the ecology of leprosy in endemic regions, a combination of geographic and molecular tools were applied for a group of 201 multibacillary patients including six multi-case families from eleven departments. The location (latitude and longitude) of patient residences were mapped. Slit skin smears and/or skin biopsies were collected and DNA was extracted. Standard agarose gel electrophoresis following a multiplex PCR-was developed for rapid and inexpensive strain typing of M. leprae based on copy numbers of two VNTR minisatellite loci 27-5 and 12-5. A SNP (C/T) in gyrA (SNP7614) was mapped by introducing a novel PCR-RFLP into an ongoing drug resistance surveillance effort. Multiple genotypes were detected combining the three molecular markers. The two frequent genotypes in Colombia were SNP7614(C)/27-5(5)/12-5(4) [C54] predominantly distributed in the Atlantic departments and SNP7614 (T)/27-5(4)/12-5(5) [T45] associated with the Andean departments. A novel genotype SNP7614 (C)/27-5(6)/12-5(4) [C64] was detected in cities along the Magdalena river which separates the Andean from Atlantic departments; a subset was further characterized showing association with a rare allele of minisatellite 23-3 and the SNP type 1 of M. leprae. The genotypes within intra-family cases were conserved. Overall, this is the first large scale study that utilized simple and rapid assay formats for identification of major strain types and their distribution in Colombia. It provides the framework for further strain type discrimination and geographic information systems as tools for tracing transmission of leprosy. PMID:23291420

  3. Administration of Mycobacterium leprae rHsp65 aggravates experimental autoimmune uveitis in mice.

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    Eliana B Marengo

    Full Text Available The 60 kDa heat shock protein family, Hsp60, constitutes an abundant and highly conserved class of molecules that are highly expressed in chronic-inflammatory and autoimmune processes. Experimental autoimmune uveitis [EAU] is a T cell mediated intraocular inflammatory disease that resembles human uveitis. Mycobacterial and homologous Hsp60 peptides induces uveitis in rats, however their participation in aggravating the disease is poorly known. We here evaluate the effects of the Mycobacterium leprae Hsp65 in the development/progression of EAU and the autoimmune response against the eye through the induction of the endogenous disequilibrium by enhancing the entropy of the immunobiological system with the addition of homologous Hsp. B10.RIII mice were immunized subcutaneously with interphotoreceptor retinoid-binding protein [IRBP], followed by intraperitoneally inoculation of M. leprae recombinant Hsp65 [rHsp65]. We evaluated the proliferative response, cytokine production and the percentage of CD4(+IL-17(+, CD4(+IFN-gamma(+ and CD4(+Foxp3(+ cells ex vivo, by flow cytometry. Disease severity was determined by eye histological examination and serum levels of anti-IRBP and anti-Hsp60/65 measured by ELISA. EAU scores increased in the Hsp65 group and were associated with an expansion of CD4(+IFN-gamma(+ and CD4(+IL-17(+ T cells, corroborating with higher levels of IFN-gamma. Our data indicate that rHsp65 is one of the managers with a significant impact over the immune response during autoimmunity, skewing it to a pathogenic state, promoting both Th1 and Th17 commitment. It seems comprehensible that the specificity and primary function of Hsp60 molecules can be considered as a potential pathogenic factor acting as a whistleblower announcing chronic-inflammatory diseases progression.

  4. Role of Subunit Exchange and Electrostatic Interactions on the Chaperone Activity of Mycobacterium leprae HSP18.

    Science.gov (United States)

    Nandi, Sandip Kumar; Panda, Alok Kumar; Chakraborty, Ayon; Sinha Ray, Sougata; Biswas, Ashis

    2015-01-01

    Mycobacterium leprae HSP18, a major immunodominant antigen of M. leprae pathogen, is a small heat shock protein. Previously, we reported that HSP18 is a molecular chaperone that prevents aggregation of different chemically and thermally stressed client proteins and assists refolding of denatured enzyme at normal temperature. We also demonstrated that it can efficiently prevent the thermal killing of E. coli at higher temperature. However, molecular mechanism behind the chaperone function of HSP18 is still unclear. Therefore, we studied the structure and chaperone function of HSP18 at normal temperature (25°C) as well as at higher temperatures (31-43°C). Our study revealed that the chaperone function of HSP18 is enhanced significantly with increasing temperature. Far- and near-UV CD experiments suggested that its secondary and tertiary structure remain intact in this temperature range (25-43°C). Besides, temperature has no effect on the static oligomeric size of this protein. Subunit exchange study demonstrated that subunits of HSP18 exchange at 25°C with a rate constant of 0.018 min(-1). Both rate of subunit exchange and chaperone activity of HSP18 is found to increase with rise in temperature. However, the surface hydrophobicity of HSP18 decreases markedly upon heating and has no correlation with its chaperone function in this temperature range. Furthermore, we observed that HSP18 exhibits diminished chaperone function in the presence of NaCl at 25°C. At elevated temperatures, weakening of interactions between HSP18 and stressed client proteins in the presence of NaCl results in greater reduction of its chaperone function. The oligomeric size, rate of subunit exchange and structural stability of HSP18 were also found to decrease when electrostatic interactions were weakened. These results clearly indicated that subunit exchange and electrostatic interactions play a major role in the chaperone function of HSP18.

  5. Pesquisa de Mycobacterium leprae em biópsias de mucosa oral por meio da reação em cadeia da polimerase Molecular detection of Mycobacterium leprae by polymerase chain reaction in oral mucosa biopsy specimens

    Directory of Open Access Journals (Sweden)

    Geraldo Gomes dos Santos

    2007-06-01

    Full Text Available FUNDAMENTOS - A hanseníase é endêmica na América do Sul, sendo responsável por 3% do total dos casos mundiais e, particularmente, no Brasil, por 85% dos casos sul-americanos. Seu agente pode ser encontrado na mucosa oral sem qualquer alteração evidente, e apenas testes laboratoriais muito sensíveis podem detectar sua presença. OBJETIVOS - Determinar se o genoma do Mycobacterium leprae pode ser encontrado pelo teste da PCR em biópsias com punch da mucosa oral de pacientes com hanseníase. MATERIAL E MÉTODOS - Realizou-se biópsia da mucosa oral normal de sete pacientes com hanseníase multibacilar. Cinco estavam em tratamento durante o estudo, e apenas um, ainda sem tratamento, teve o diagnóstico confirmado pela hematoxilina-eosina e coloração de Fite-Faraco para M. leprae. As peças foram incluídas em parafina e submetidas à PCR para pesquisa de M. leprae. RESULTADOS - Seis dos sete casos foram positivos para M. leprae, e um para Mycobacterium sp., demonstrando-se alta sensibilidade e especificidade do método. CONCLUSÃO - A PCR é método rápido, fácil e confiável para a investigação de rotina da infecção por micobactéria, mesmo quando a doença ainda é assintomática. O diagnóstico pode ser obtido a partir de simples biópsia ambulatorial.BACKGROUND - Hansen's disease is endemic in South America, which accounts for 3% of total world cases, and particularly in Brazil, which accounts for 85% of all South American cases. The bacteria can be found in the oral mucosa with no evident signs of infection, and only very sensitive laboratory assays can detect its presence. OBJECTIVES - The aim of this study was to ascertain if the M. leprae genome can be detected by PCR in small punch biopsy specimens from the oral mucosa of patients with Hansen's disease. METHODS - The normal oral mucosas of seven multibacillary Hansen's disease patients were biopsied. Five of them were under treatment at the time of the study. Diagnosis of

  6. Zinc blocks SOS-induced antibiotic resistance via inhibition of RecA in Escherichia coli.

    Directory of Open Access Journals (Sweden)

    Bryan E Bunnell

    Full Text Available Zinc inhibits the virulence of diarrheagenic E. coli by inducing the envelope stress response and inhibiting the SOS response. The SOS response is triggered by damage to bacterial DNA. In Shiga-toxigenic E. coli, the SOS response strongly induces the production of Shiga toxins (Stx and of the bacteriophages that encode the Stx genes. In E. coli, induction of the SOS response is accompanied by a higher mutation rate, called the mutator response, caused by a shift to error-prone DNA polymerases when DNA damage is too severe to be repaired by canonical DNA polymerases. Since zinc inhibited the other aspects of the SOS response, we hypothesized that zinc would also inhibit the mutator response, also known as hypermutation. We explored various different experimental paradigms to induce hypermutation triggered by the SOS response, and found that hypermutation was induced not just by classical inducers such as mitomycin C and the quinolone antibiotics, but also by antiviral drugs such as zidovudine and anti-cancer drugs such as 5-fluorouracil, 6-mercaptopurine, and azacytidine. Zinc salts inhibited the SOS response and the hypermutator phenomenon in E. coli as well as in Klebsiella pneumoniae, and was more effective in inhibiting the SOS response than other metals. We then attempted to determine the mechanism by which zinc, applied externally in the medium, inhibits hypermutation. Our results show that zinc interferes with the actions of RecA, and protects LexA from RecA-mediated cleavage, an early step in initiation of the SOS response. The SOS response may play a role in the development of antibiotic resistance and the effect of zinc suggests ways to prevent it.

  7. qPCR detection of Mycobacterium leprae in biopsies and slit skin smear of different leprosy clinical forms

    Directory of Open Access Journals (Sweden)

    Michelle de Campos Soriani Azevedo

    2017-01-01

    Full Text Available Leprosy, whose etiological agent is Mycobacterium leprae, is a chronic infectious disease that mainly affects the skin and peripheral nervous system. The diagnosis of leprosy is based on clinical evaluation, whereas histopathological analysis and bacilloscopy are complementary diagnostic tools. Quantitative PCR (qPCR, a current useful tool for diagnosis of infectious diseases, has been used to detect several pathogens including Mycobacterium leprae. The validation of this technique in a robust set of samples comprising the different clinical forms of leprosy is still necessary. Thus, in this study samples from 126 skin biopsies (collected from patients on all clinical forms and reactional states of leprosy and 25 slit skin smear of leprosy patients were comparatively analyzed by qPCR (performed with primers for the RLEP region of M. leprae DNA and routine bacilloscopy performed in histological sections or in slit skin smear. Considering clinical diagnostic as the gold standard, 84.9% of the leprosy patients were qPCR positive in skin biopsies, resulting in 84.92% sensitivity, with 84.92 and 61.22% positive (PPV and negative (NPV predictive values, respectively. Concerning bacilloscopy of histological sections (BI/H, the sensitivity was 80.15% and the PPV and NPV were 80.15 and 44.44%, respectively. The concordance between qPCR and BI/H was 87.30%. Regarding the slit skin smear, 84% of the samples tested positive in the qPCR. Additionally, qPCR showed 100% specificity, since all samples from different mycobacteria, from healthy individuals, and from other granulomatous diseases presented negative results. In conclusion, the qPCR technique for detection of M. leprae using RLEP primers proved to be specific and sensitive, and qPCR can be used as a complementary test to diagnose leprosy irrespective of the clinical form of disease.

  8. qPCR detection of Mycobacterium leprae in biopsies and slit skin smear of different leprosy clinical forms.

    Science.gov (United States)

    Azevedo, Michelle de Campos Soriani; Ramuno, Natália Mortari; Fachin, Luciana Raquel Vincenzi; Tassa, Mônica; Rosa, Patrícia Sammarco; Belone, Andrea de Faria Fernandes; Diório, Suzana Madeira; Soares, Cleverson Teixeira; Garlet, Gustavo Pompermaier; Trombone, Ana Paula Favaro

    Leprosy, whose etiological agent is Mycobacterium leprae, is a chronic infectious disease that mainly affects the skin and peripheral nervous system. The diagnosis of leprosy is based on clinical evaluation, whereas histopathological analysis and bacilloscopy are complementary diagnostic tools. Quantitative PCR (qPCR), a current useful tool for diagnosis of infectious diseases, has been used to detect several pathogens including Mycobacterium leprae. The validation of this technique in a robust set of samples comprising the different clinical forms of leprosy is still necessary. Thus, in this study samples from 126 skin biopsies (collected from patients on all clinical forms and reactional states of leprosy) and 25 slit skin smear of leprosy patients were comparatively analyzed by qPCR (performed with primers for the RLEP region of M. leprae DNA) and routine bacilloscopy performed in histological sections or in slit skin smear. Considering clinical diagnostic as the gold standard, 84.9% of the leprosy patients were qPCR positive in skin biopsies, resulting in 84.92% sensitivity, with 84.92 and 61.22% positive (PPV) and negative (NPV) predictive values, respectively. Concerning bacilloscopy of histological sections (BI/H), the sensitivity was 80.15% and the PPV and NPV were 80.15 and 44.44%, respectively. The concordance between qPCR and BI/H was 87.30%. Regarding the slit skin smear, 84% of the samples tested positive in the qPCR. Additionally, qPCR showed 100% specificity, since all samples from different mycobacteria, from healthy individuals, and from other granulomatous diseases presented negative results. In conclusion, the qPCR technique for detection of M. leprae using RLEP primers proved to be specific and sensitive, and qPCR can be used as a complementary test to diagnose leprosy irrespective of the clinical form of disease. Copyright © 2016 Sociedade Brasileira de Infectologia. Published by Elsevier Editora Ltda. All rights reserved.

  9. Single nucleotide polymorphism-based molecular typing of M. leprae from multicase families of leprosy patients and their surroundings to understand the transmission of leprosy.

    Science.gov (United States)

    Turankar, R P; Lavania, M; Chaitanya, V S; Sengupta, U; Darlong, J; Darlong, F; Siva Sai, K S R; Jadhav, R S

    2014-03-01

    The exact mode of transmission of leprosy is not clearly understood; however, many studies have demonstrated active transmission of leprosy around a source case. Families of five active leprosy cases and their household contacts were chosen from a high endemic area in Purulia. Fifty-two soil samples were also collected from different areas of their houses. DNA was extracted from slit-skin smears (SSS) and soil samples and the Mycobacterium leprae-specific RLEP (129 bp) region was amplified using PCR. Molecular typing of M. leprae was performed for all RLEP PCR-positive samples by single nucleotide polymorphism (SNP) typing and confirmation by DNA sequencing. SSS of these five patients and six out of the total 28 contacts were PCR positive for RLEP whereas 17 soil samples out of 52 showed the presence of M. leprae DNA. SNP typing of M. leprae from all RLEP PCR-positive subjects (patients and smear-positive contacts) and 10 soil samples showed the SNP type 1 genotype. M. leprae DNA from the five leprosy patients and the six contacts was further subtyped and the D subtype was noted in all patients and contacts, except for one contact where the C subtype was identified. Typing followed by subtyping of M. leprae clearly revealed that either the contacts were infected by the patients or both patients and contacts had the same source of infection. It also revealed that the type of M. leprae in the soil in the inhabited areas where patients resided was also of the same type as that found in patients. © 2013 The Authors Clinical Microbiology and Infection © 2013 European Society of Clinical Microbiology and Infectious Diseases.

  10. UltramicroELISA para la detección de anticuerpos IgM anti M. leprae UltramicroELISA assay for the detection of human IgM antibodies to M. leprae

    Directory of Open Access Journals (Sweden)

    José Laferte

    1991-12-01

    Full Text Available La disponibilidad del sistema Ultramicroanalítico (SUMA y de un antígeno especie-específico del M. leprae obtenido mediante síntesis química, permitió la normalización y validación de un ultramicroELISA para la detección de anticuerpos IgM específicos a esta micobacteria. El análisis de 433 sueros de banco de sangre y 265 sueros usados para validar el método y clasificados en un grupo control de donantes de banco de sangre (100, un grupo de pacientes tuberculosos (50, un grupo de enfermos de lepra (65 y un grupo de contactos de estos enfermos (50, mostró la especificidad del ensayo para evidenciar la infección con el M. leprae. Los resultados obtenidos del estudio adicional de 140 muestras de suero de contactos de enfermos estuvieron estrechamente correlacionados (r = 0,98 con los resultados obtenidos por la técnica de microELISA convencional. La utilización del SUMA no solo permite un notable ahorro de reactivos si no además facilita la lectura, cálculo, validación y almacenamiento automático de los resultados.The availability of an ultramicroanalitic system (SUMA and specie-specific antigen of M. leprae obtained by chemical synthesis, have made possible the standardization and validation of an ultramicroELISA assay for detecting specific human IgM antibodies to this mycobacterium. The specificity of this test to demonstrate the infection with M. leprae was corroborated through a screening of 433 blood bank serum samples and other 265 from diferent groups (100, control group, 50 tuberculosis patients, 65 leprosy patients, 50 from household. The results obtained in the aditional study of 140 household sero showed a high correlation (r = 0.98 with the conventional microELISA method. The use of SUMA allows saving reagents and time since sample handling, plate reading, print out and storing the data are computer assisted.

  11. Comparative evolution of the recA gene of surface and deep subsurface microorganisms (an evolutionary clock of intermediate rate). Final report

    Energy Technology Data Exchange (ETDEWEB)

    Miller, R.V.

    1998-04-01

    Because of the ability of the recA protein product to maintain both DNA integrity and increase genetic diversity, this gene may be essential to the survival of microorganisms following the damaging effects of numerous environmental stresses such as exposure to solar UV radiation, exposure to gamma radiation, starvation, and changing environments. While the various activities and amino-acid sequence of recA have been highly conserved among the eubacteria and archaea, little is known as to whether a strict structure-function relationship has been conserved. In other words, are the same regions of this highly plastic, functionally heterogeneous protein involved in the same catalytic capacities throughout the bacterial kingdom? While it is reasonable to assume that this type of conservation has also occurred, we felt it necessary to test the assumption by demonstrating that mutations in different genera of bacteria which eliminate similar functions (i.e., lead to similar phenotypes) are caused by changes in the amino-acid sequence in the same regions of their recA proteins. Therefore, we located the changes in nucleotide sequence in two recA mutants of P. aeruginosa which displayed mutant phenotypes in recombination and UV resistance. Our assumption was that if structure-function relationships held, these mutations would be found in areas already identified as essential for the function of the E. coli recA protein.

  12. La recaída en fumadores que dejan de fumar con un tratamiento psicológico: ¿una cuestión de sexo?

    Directory of Open Access Journals (Sweden)

    Úrsula Martínez

    Full Text Available Resumen Aunque contamos con tratamientos eficaces para la dependencia del tabaco, la recaída continúa siendo un fenómeno frecuente en aquellos que consiguen dejar de fumar. El objetivo del presente estudio fue analizar si existían diferencias en función del sexo en la curva de recaída en hombres y mujeres que participaron en un tratamiento psicológico cognitivo-conductual para dejar de fumar. La muestra estuvo formada por 266 personas (59.8 % mujeres que dejaron de fumar al finalizar el tratamiento. Los resultados indicaron que a medida que se incrementaba el tiempo transcurrido tras la intervención, disminuía el porcentaje de participantes abstinentes. No obstante, no se encuentraron diferencias significativas entre hombres y mujeres en la curva de recaída, lo cual podría explicarse por la inclusión en el tratamiento de técnicas psicológicas eficaces para el afrontamiento de algunas de las principales causas de la recaída (e.g., estado de ánimo negativo. Dada la importancia que tiene la recaída en el proceso de dejar de fumar, es necesario profundizar en los distintos determinantes que la desencadenan.

  13. Avances en el tratamiento de los pacientes con Leucemia Promielocítica Aguda en Recaída

    Directory of Open Access Journals (Sweden)

    A. Sanz Miguel

    2006-04-01

    Full Text Available Hasta la demostración de la actividad sobresaliente del trióxido de arsénico (ATO en recaídas de pacientes con leucemia promielocítica (LPA, el tratamiento de rescate en esta enfermedad consistía en la readministración de ácido holo-trans retinóico (ATRA y quimioterapia para inducir la remisión, generalmente conteniendo citarabina a altas dosis, seguido de consolidación y/o trasplante de progenitores hematopoyéticos (TPH.

  14. Manejo terapéutico en reinfección postratamiento, recaídas no respondedores

    OpenAIRE

    León de Mezerville-Cantillo

    2008-01-01

    A pesar del uso universal de la vacuna, la hepatitis B sigue siendo un problema de salud pública en diferentes áreas del mundo. Aún cuando se consiga una supresión eficaz de la replicación viral mediante tratamiento con fármacos, existe la potencial reactivación al suspender el mismo. El enfoque terapéutico para los no respondedores o recaídas post tratamiento, se podría llevar a cabo con combinaciones de medicamentos o el uso de otro tipo de fármaco con mayor potencia.

  15. Type 1- and type 2-like lesional skin-derived Mycobacterium leprae-responsive T cell clones are characterized by coexpression of IFN-gamma/TNF-alpha and IL-4/IL-5/IL-13, respectively

    NARCIS (Netherlands)

    Verhagen, C. E.; van der Pouw Kraan, T. C.; Buffing, A. A.; Chand, M. A.; Faber, W. R.; Aarden, L. A.; Das, P. K.

    1998-01-01

    In an earlier study, we generated a large number of Mycobacterium leprae-responsive and M. leprae-nonresponsive T cell clones (TCC) from the lesional skin of immunologic unstable borderline leprosy patients. In that study, we divided TCC into type 1- and type 2-like on the basis of their IFN-gamma

  16. Circumscription of the genus Lepra, a recently resurrected genus to accommodate the "Variolaria"-group of Pertusaria sensu lato (Pertusariales, Ascomycota).

    Science.gov (United States)

    Wei, Xinli; Schmitt, Imke; Hodkinson, Brendan; Flakus, Adam; Kukwa, Martin; Divakar, Pradeep K; Kirika, Paul M; Otte, Jürgen; Meiser, Anjuli; Lumbsch, H Thorsten

    2017-01-01

    Pertusarialean lichens include more than 300 species belonging to several independent phylogenetic lineages. Only some of these phylogenetic clades have been comprehensively sampled for molecular data, and formally described as genera. Here we present a taxonomic treatment of a group of pertusarialean lichens formerly known as "Pertusaria amara-group", "Monomurata-group", or "Variolaria-group", which includes widespread and well-known taxa such as P. amara, P. albescens, or P. ophthalmiza. We generated a 6-locus data set with 79 OTUs representing 75 species. The distinction of the Variolaria clade is supported and consequently, the resurrection of the genus Lepra is followed. Thirty-five new combinations into Lepra are proposed and the new species Lepra austropacifica is described from mangroves in the South Pacific. Lepra is circumscribed to include species with disciform ascomata, a weakly to non-amyloid hymenial gel, strongly amyloid asci without clear apical amyloid structures, containing 1 or 2, single-layered, thin-walled ascospores. Chlorinated xanthones are not present, but thamnolic and picrolichenic acids occur frequently, as well as orcinol depsides. Seventy-one species are accepted in the genus. Although the distinction of the genus from Pertusaria is strongly supported, the relationships of Lepra remain unresolved and the genus is tentatively placed in Pertusariales incertae sedis.

  17. Identification of Bacillus Probiotics Isolated from Soil Rhizosphere Using 16S rRNA, recA, rpoB Gene Sequencing and RAPD-PCR.

    Science.gov (United States)

    Mohkam, Milad; Nezafat, Navid; Berenjian, Aydin; Mobasher, Mohammad Ali; Ghasemi, Younes

    2016-03-01

    Some Bacillus species, especially Bacillus subtilis and Bacillus pumilus groups, have highly similar 16S rRNA gene sequences, which are hard to identify based on 16S rDNA sequence analysis. To conquer this drawback, rpoB, recA sequence analysis along with randomly amplified polymorphic (RAPD) fingerprinting was examined as an alternative method for differentiating Bacillus species. The 16S rRNA, rpoB and recA genes were amplified via a polymerase chain reaction using their specific primers. The resulted PCR amplicons were sequenced, and phylogenetic analysis was employed by MEGA 6 software. Identification based on 16S rRNA gene sequencing was underpinned by rpoB and recA gene sequencing as well as RAPD-PCR technique. Subsequently, concatenation and phylogenetic analysis showed that extent of diversity and similarity were better obtained by rpoB and recA primers, which are also reinforced by RAPD-PCR methods. However, in one case, these approaches failed to identify one isolate, which in combination with the phenotypical method offsets this issue. Overall, RAPD fingerprinting, rpoB and recA along with concatenated genes sequence analysis discriminated closely related Bacillus species, which highlights the significance of the multigenic method in more precisely distinguishing Bacillus strains. This research emphasizes the benefit of RAPD fingerprinting, rpoB and recA sequence analysis superior to 16S rRNA gene sequence analysis for suitable and effective identification of Bacillus species as recommended for probiotic products.

  18. Mycobacterium leprae in the periodontium, saliva and skin smears of leprosy patients = Mycobacterium leprae no periodonto, saliva e raspados intradérmicos de sujeitos com hanseníase

    Directory of Open Access Journals (Sweden)

    Abdalla, Ligia Fernandes

    2010-01-01

    Full Text Available Objetivo: verificar através da baciloscopia e da reação em cadeia da polimerase (PCR a presença do M. leprae no periodonto, saliva e raspados intradérmicos de pacientes com hanseníase. Metodologia: Realizou-se um estudo transversal do tipo detecção de casos numa instituição referência de hanseníase no Amazonas. Resultados: Foram avaliados 48 pacientes, sendo 15 multibacilares (MB e 33 paucibacilares (PB. Os pacientes MB tiveram o diagnóstico confirmado pela baciloscopia e PCR dos raspados intradérmicos, enquanto que 16 (48,5% dos PB foram positivos apenas na PCR. Quatro pacientes PB negativos na PCR de raspados intradérmicos foram positivos no periodonto e na saliva, 1 positivo na saliva e 2 no periodonto. Nenhuma amostra do periodonto e da saliva foi positiva na baciloscopia. Conclusão: Não houve relação entre a doença periodontal e a presença do M. leprae; a baciloscopia não mostrou ser uma técnica eficiente para análise da saliva e periodonto; a técnica de PCR de raspado dérmico mostrou ser um método mais eficaz no diagnóstico dos PB do que a baciloscopia; a positividade da PCR para detecção do M. leprae nos PB pode ser aumentada coletando raspado intradérmico, periodonto e saliva

  19. Interruption of persistent exposure to leprosy combined or not with recent BCG vaccination enhances the response to Mycobacterium leprae specific antigens.

    Science.gov (United States)

    de Carvalho, Fernanda Marques; Rodrigues, Luciana Silva; Duppre, Nádia Cristina; Alvim, Iris Maria Peixoto; Ribeiro-Alves, Marcelo; Pinheiro, Roberta Olmo; Sarno, Euzenir Nunes; Pessolani, Maria Cristina Vidal; Pereira, Geraldo Moura Batista

    2017-05-01

    Household contacts of multibacillary leprosy patients (HCMB) constitute the group of individuals at the highest risk of developing leprosy. Early diagnosis and treatment of their index cases combined with Bacille Calmette-Guerin (BCG) immunization remain important strategies adopted in Brazil to prevent HCMB from evolving into active disease. In the present study, we assessed the impact of these measures on the immune response to Mycobacterium leprae in HCMB. Peripheral blood mononuclear cells (PBMC) from HCMB (n = 16) were obtained at the beginning of leprosy index case treatment (T0). At this time point, contacts were vaccinated (n = 13) or not (n = 3) in accordance with their infancy history of BCG vaccination and PBMCs were recollected at least 6 months later (T1). As expected, a significant increase in memory CD4 and CD8 T cell frequencies responsive to M. leprae whole-cell sonicate was observed in most contacts. Of note, higher frequencies of CD4+ T cells that recognize M. leprae specific epitopes were also detected. Moreover, increased production of the inflammatory mediators IL1-β, IL-6, IL-17, TNF, IFN-γ, MIP1-β, and MCP-1 was found at T1. Interestingly, the increment in these parameters was observed even in those contacts that were not BCG vaccinated at T0. This result reinforces the hypothesis that the continuous exposure of HCMB to live M. leprae down regulates the specific cellular immune response against the pathogen. Moreover, our data suggest that BCG vaccination of HCMB induces activation of T cell clones, likely through "trained immunity", that recognize M. leprae specific antigens not shared with BCG as an additional protective mechanism besides the expected boost in cell-mediated immunity by BCG homologues of M. leprae antigens.

  20. Mycobacterium leprae in the periodontium, saliva and skin smears of leprosy patients = Mycobacterium leprae no periodonto, saliva e raspados intradérmicos de sujeitos com hanseníase

    OpenAIRE

    Abdalla, Ligia Fernandes

    2010-01-01

    Objetivo: verificar através da baciloscopia e da reação em cadeia da polimerase (PCR) a presença do M. leprae no periodonto, saliva e raspados intradérmicos de pacientes com hanseníase. Metodologia: Realizou-se um estudo transversal do tipo detecção de casos numa instituição referência de hanseníase no Amazonas. Resultados: Foram avaliados 48 pacientes, sendo 15 multibacilares (MB) e 33 paucibacilares (PB). Os pacientes MB tiveram o diagnóstico confirmado pela baciloscopia e PCR dos raspados ...

  1. Computational Modelling of Dapsone Interaction With Dihydropteroate Synthase in Mycobacterium leprae; Insights Into Molecular Basis of Dapsone Resistance in Leprosy.

    Science.gov (United States)

    Chaitanya V, Sundeep; Das, Madhusmita; Bhat, Pritesh; Ebenezer, Mannam

    2015-10-01

    The molecular basis for determination of resistance to anti-leprosy drugs is the presence of point mutations within the genes of Mycobacterium leprae (M. leprae) that encode active drug targets. The downstream structural and functional implications of these point mutations on drug targets were scarcely studied. In this study, we utilized computational tools to develop native and mutant protein models for 5 point mutations at codon positions 53 and 55 in 6-hydroxymethyl-7, 8-dihydropteroate synthase (DHPS) of M. leprae, an active target for dapsone encoded by folp1 gene, that confer resistance to dapsone. Molecular docking was performed to identify variations in dapsone interaction with mutant DHPS in terms of hydrogen bonding, hydrophobic interactions, and energy changes. Schrodinger Suite 2014-3 was used to build homology models and in performing molecular docking. An increase in volume of the binding cavities of mutant structures was noted when compared to native form indicating a weakening in interaction (60.7 Å(3) in native vs. 233.6 Å(3) in Thr53Ala, 659.9 Å(3) in Thr53Ile, 400 Å(3) for Thr53Val, 385 Å(3) for Pro55Arg, and 210 Å(3) for Pro55Leu). This was also reflected by changes in hydrogen bonds and decrease in hydrophobic interactions in the mutant models. The total binding energy (ΔG) decreased significantly in mutant forms when compared to the native form (-51.92 Kcal/mol for native vs. -35.64, -35.24, -46.47, -47.69, and -41.36 Kcal/mol for mutations Thr53Ala, Thr53Ile, Thr53Val, Pro55Arg, and Pro55Leu, respectively. In brief, this analysis provided structural and mechanistic insights to the degree of dapsone resistance contributed by each of these DHPS mutants in leprosy. © 2015 Wiley Periodicals, Inc.

  2. Thio and hydrogen peroxide modofication of recA induction in UV-irradiated wild-type and catalase-deficient Escherichia coli K12

    International Nuclear Information System (INIS)

    Claycamp, H.G.; Kam-Kuen Ho; DeRose, C.; Iowa Univ., Iowa City, IA

    1990-01-01

    Induction of recA in Escherichia coli, monitored as β-D-galactosidase activity in recA-lacZ fusion strains, was shown to be elevated and prolonged by dithiothreitol (DTT) treatment after far-UV radiation. Pretreatment of UV-irradiated coltures using DTT led to a shortened recA response and little increase of β-Gal yield. Similar studies were performed using a catalase-deficient recA-lacZ strain in which the major feature was elevated levels of recA-lacZ induction. Catalase activity in UV-irradiated wild-type cells was reduced by DTT treatment to levels as low as in a katE mutant strain, leading to similar recA-lacZ induction patterns between the strains. Neither DTT nor H 2 O 2 treatment of cells could induce significant recA transcription in the absence of UV-radiation, implying that both agents modify recA activity primarily by interfering with repair of recA-inducing DNA lesions. The results confirm previous studies suggesting that modification of DNA repair is probably a significant portion of thiol radiation protection. (author). 36 refs.; 7 figs.; 1 tab

  3. High-temperature piezoelectric crystals ReCa4O(BO3)3: a review.

    Science.gov (United States)

    Yu, Fapeng; Hou, Shuai; Zhao, Xian; Zhang, Shujun

    2014-08-01

    High-temperature sensors are desirable for structural health monitoring and/or nondestructive evaluation of next-generation turbines, more efficient jet engines, and the furnace components of electrical power plants. Of all the investigated high-temperature piezoelectric materials, rare-earth calcium oxyborate crystals ReCa4O(BO3)3 (ReCOB, Re: rare-earth) exhibit attractive advantages for high-temperature piezoelectric sensing. In this paper, the electroelastic properties of different ReCOB piezoelectric crystals are investigated. The crosstalk between various vibration modes are discussed, from which the optimized crystal cuts are achieved. Furthermore, temperature dependences of the electrical resistivity, dielectric, elastic, piezoelectric, and electromechanical properties of ReCOB crystals are studied. Finally, the thermal properties, including thermal expansion, specific heat, and thermal conductivity at elevated temperatures are studied and compared with commercially available high-temperature piezoelectric crystals.

  4. Rare-Earth Calcium Oxyborate Piezoelectric Crystals ReCa4O(BO33: Growth and Piezoelectric Characterizations

    Directory of Open Access Journals (Sweden)

    Fapeng Yu

    2014-07-01

    Full Text Available Rare-earth calcium oxyborate crystals, ReCa4O(BO33 (ReCOB, Re = Er, Y, Gd, Sm, Nd, Pr, and La , are potential piezoelectric materials for ultrahigh temperature sensor applications, due to their high electrical resistivity at elevated temperature, high piezoelectric sensitivity and temperature stability. In this paper, different techniques for ReCOB single-crystal growth are introduced, including the Bridgman and Czochralski pulling methods. Crystal orientations and the relationships between the crystallographic and physical axes of the monoclinic ReCOB crystals are discussed. The procedures for dielectric, elastic, electromechanical and piezoelectric property characterization, taking advantage of the impedance method, are presented. In addition, the maximum piezoelectric coefficients for different piezoelectric vibration modes are explored, and the optimized crystal cuts free of piezoelectric cross-talk are obtained by rotation calculations.

  5. Expression of RecA and cell-penetrating peptide (CPP) fusion protein in bacteria and in mammalian cells.

    Science.gov (United States)

    Chang, Xiubao; Hou, Yuexian

    2018-01-01

    Genome editing is a powerful tool to modify a specific gene and to correct a disease-causing mutation. Recently developed new techniques, such as zinc-finger nucleases (ZFNs), transcription activator-like effector nucleases (TALEN) and clustered regularly interspaced short palindromic repeats/Cas9 (CRISPR/Cas9), significantly facilitate the progression in this field. However, mutations associated with the double strand DNA breaks (DSBs) introduced by these systems hampered their direct usage in clinic. In order to prevent the mutations caused by DSBs, we have designed a novel mean to induce homology-directed recombination (HDR) without DSBs, i.e., the fusion protein of RecA with cell-penetrating peptide (CPP). The involvement of RecA in these fusion proteins will play important roles in formation of the nucleoprotein filament with single strand DNA (ssDNA) in vitro and promoting HDR in vivo ; whereas the involvement of CPP in these fusion proteins will mainly play a role in facilitating cellular intake/uptake of the nucleoprotein filaments. Our results indicated that certain amount of the fusion proteins expressed in bacteria is in soluble fraction, whereas majority of the fusion proteins expressed in baby hamster kidney (BHK) cells is in soluble fraction. Interestingly, expression of these fusion proteins in bacteria completely blocked cell growth, whereas expression of them in BHK cells significantly inhibited cell growth, implying that these fusion proteins may bind to ssDNA regions, such as ssDNA regions in DNA replication forks, and inhibit cell growth. These results suggest that we have functional RecA.CPP fusion proteins ready to test our novel idea of inducing HDR without DSB.

  6. El estigma en la representación social de la lepra

    Directory of Open Access Journals (Sweden)

    Alexis Romero-Salazar

    Full Text Available La representación social de la lepra en el paciente activo bajo control se constituye de elementos provenientes del discurso médico y del discurso que, en torno a la afección, circula en la sociedad. Tal sincretismo se expresa, en cada contexto, de una manera específica, sobre los componentes de la representación - nociones, creencias, actitudes y conductas - haciendo de ella una realidad compleja. En razón del peso que tienen las creencias, cuya profundidad y arraigo están vinculadas a la antigüedad de la enfermedad y a las prácticas confinatorias vigentes hasta épocas recientes, sería comprensible el predominio de los elementos ligados al sentido común o saber popular. No obstante, siendo la representación una construcción colectiva, sus rasgos fundamentales estarían definidos por las específicas características del grupo. Al respecto, los principales hallazgos de la investigación se refieren, primero, al escaso conocimiento de los pacientes activos en el Servicio de Dermatología Sanitaria de Maracaibo (Venezuela en torno a la enfermedad y a la existencia de creencias y actitudes inadecuadas para los programas de control: segundo, al predominio, en la dimensión cognoscitiva de la representación, de nociones y creencias en relación al papel de la suerte o del destino, de la promiscuidad sexual o de la herencia en la transmisión; tercero, a la tendencia a la participación del paciente en la reproducción de un estereotipo descalificador y al ocultamiento, aunado a un exagerado miedo al rechazo.

  7. Administration of M. leprae Hsp65 interferes with the murine lupus progression.

    Directory of Open Access Journals (Sweden)

    Eliana B Marengo

    Full Text Available The heat shock protein [Hsp] family guides several steps during protein synthesis, are abundant in prokaryotic and eukaryotic cells, and are highly conserved during evolution. The Hsp60 family is involved in assembly and transport of proteins, and is expressed at very high levels during autoimmunity or autoinflammatory phenomena. Here, the pathophysiological role of the wild type [WT] and the point mutated K(409A recombinant Hsp65 of M. leprae in an animal model of Systemic Lupus Erythematosus [SLE] was evaluated in vivo using the genetically homogeneous [NZBxNZW]F(1 mice. Anti-DNA and anti-Hsp65 antibodies responsiveness was individually measured during the animal's life span, and the mean survival time [MST] was determined. The treatment with WT abbreviates the MST in 46%, when compared to non-treated mice [p<0.001]. An increase in the IgG2a/IgG1 anti-DNA antibodies ratio was also observed in animals injected with the WT Hsp65. Incubation of BALB/c macrophages with F(1 serum from WT treated mice resulted in acute cell necrosis; treatment of these cells with serum from K(409A treated mice did not cause any toxic effect. Moreover, the involvement of WT correlates with age and is dose-dependent. Our data suggest that Hsp65 may be a central molecule intervening in the progression of the SLE, and that the point mutated K(409A recombinant immunogenic molecule, that counteracts the deleterious effect of WT, may act mitigating and delaying the development of SLE in treated mice. This study gives new insights into the general biological role of Hsp and the significant impact of environmental factors during the pathogenesis of this autoimmune process.

  8. Leprosy and the elusive M. leprae: colonial and imperial medical exchanges in the nineteenth century A lepra e o evasivo M. leprae: a troca de informações médicas nos períodos colonial e imperial do século XIX

    Directory of Open Access Journals (Sweden)

    Jo Robertson

    2003-01-01

    Full Text Available In the 1800s, humoral understandings of leprosy successively give way to disease models based on morbid anatomy, physiopathology, and bacteriology. Linkages between these disease models were reinforced by the ubiquitous seed/soil metaphor deployed both before and after the identification of M. leprae. While this metaphor provided a continuous link between medical descriptions, Henry Vandyke Carter's On leprosy (1874 marks a convergence of different models of disease. Simultaneously, this metaphor can be traced in popular and medical debates in the late nineteenth century, accompanying fears of a resurgence of leprosy in Europe. Later the mapping of the genome ushers in a new model of disease but, ironically, while leprosy research draws its logic from a view of the world in which a seed and soil metaphor expresses many different aspects of the activity of the disease, the bacillus itself continues to be unreceptive to cultivation.No século XIX, abordagens humorais da lepra deram origem a sucessivos modelos da doença baseados na anatomia patológica, na fisiopatologia e na bacteriologia. As relações entre esses modelos da doença foram reforçadas pela onipresente metáfora 'da semente e do solo', difundida tanto antes quanto depois da identificação do M. leprae. À época em que a metáfora fornecia um elo de ligação contínuo entre as várias descrições médicas da doença, Henry Vandyke Carter publicava On leprosy (1874, estabelecendo uma convergência de seus diferentes modelos. Simultaneamente, a metáfora se fazia presente nos debates médicos e populares de fins do século XIX, juntamente com o medo do surgimento da lepra na Europa. Mais recentemente, o mapeamento do genoma humano determinou a formulação de um novo modelo para a doença. Mas, ironicamente, enquanto as pesquisas concernentes a ela se apóiam numa visão de mundo em que a metáfora da semente e do solo ainda expressa diferentes aspectos da ação da doença, o pr

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

    Science.gov (United States)

    Phelan, Jody; Maitra, Arundhati; McNerney, Ruth; Nair, Mridul; Gupta, Antima; Coll, Francesc; Pain, Arnab; Bhakta, Sanjib; Clark, Taane G

    2015-09-01

    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 6.02Mb 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. Copyright © 2015 Asian-African Society for Mycobacteriology. Published by Elsevier Ltd. All rights reserved.

  10. Lepra en la infancia: Desafío diagnóstico Leprosy in childhood: Diagnostic challenge

    Directory of Open Access Journals (Sweden)

    AE Táquez

    2011-12-01

    Full Text Available La lepra en la infancia cursa con una diversidad de manifestaciones clínicas e histopatológicas, que hacen necesario un minucioso examen cutáneo en todo niño, que presente lesiones dermatológicas sugestivas y una fuente infecciosa sospechosa. Para un oportuno diagnóstico es indispensable que el médico tenga siempre presente la enfermedad, así como destreza al realizar el examen clínico, ya que muchas lesiones cutáneas suelen ser asintomáticas y con frecuencia simulan otros cuadros dermatológicos. Presentamos tres casos de pacientes erróneamente diagnosticados, tratados por otras dermatosis y quienes finalmente estaban afectados de lepra.Leprosy in childhood course with a diversity of clinical and histopathological signs that make necessary a detailed cutaneous inspection in every child that presents suggestive dermatological lesions, have had a suspected infectious contact. For an appropriate diagnosis, is very important to keep in mind the disease, as well as medical clinical skill because many skin and nerve lesions can be asymptomatic and frequently look like other dermatological pathology. The authors report three cases of children misdiagnosed and treated for other dermatosis and whom finally were found leprosy´s patients.

  11. 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.

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

    Directory of Open Access Journals (Sweden)

    Jody Phelan

    2015-01-01

    Full Text Available 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 emb B 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.

  13. Two cases of leprosy from Žatec (Bohemia), dated to the turn of the 12th century and confirmed by DNA analysis for Mycobacterium leprae

    Czech Academy of Sciences Publication Activity Database

    Likovský, Jakub; Urbanová, M.; Hájek, Martin; Černý, Viktor; Čech, Petr

    2006-01-01

    Roč. 33, č. 9 (2006), s. 1276-1283 ISSN 0305-4403 Institutional research plan: CEZ:AV0Z80020508 Keywords : Leprosy * mediaeval * aDNA * Mycobacterium leprae * Paleopathology Subject RIV: AC - Archeology, Anthropology, Ethnology Impact factor: 1.322, year: 2006

  14. Evaluation of Mycobacterium leprae antigens in the monitoring of a dapsone-based chemotherapy of previously untreated lepromatous patients in Cebu, Philippines

    NARCIS (Netherlands)

    Klatser, P. R.; de Wit, M. Y.; Fajardo, T. T.; Cellona, R. V.; Abalos, R. M.; de la Cruz, E. C.; Madarang, M. G.; Hirsch, D. S.; Douglas, J. T.

    1989-01-01

    Thirty-five previously untreated lepromatous patients receiving dapsone-based therapy were monitored throughout their 5-year period of treatment by serology and by pathology. Sequentially collected sera were used to evaluate the usefulness of four Mycobacterium leprae antigens as used in ELISA to

  15. Mycobacteria contain two groEL genes: the second Mycobacterium leprae groEL gene is arranged in an operon with groES

    NARCIS (Netherlands)

    Rinke de Wit, T. F.; Bekelie, S.; Osland, A.; Miko, T. L.; Hermans, P. W.; van Soolingen, D.; Drijfhout, J. W.; Schöningh, R.; Janson, A. A.; Thole, J. E.

    1992-01-01

    In contrast to other bacterial species, mycobacteria were thus far considered to contain groEL and groES genes that are present on separate loci on their chromosomes, Here, by screening a Mycobacterium leprae lambda gt11 expression library with serum from an Ethiopian lepromatous leprosy patient,

  16. Genotyping of Mycobacterium leprae present on Ziehl-Neelsen-stained microscopic slides and in skin biopsy samples from leprosy patients in different geographic regions of Brazil

    Directory of Open Access Journals (Sweden)

    Amanda Nogueira Brum Fontes

    2012-12-01

    Full Text Available We analysed 16 variable number tandem repeats (VNTR and three single-nucleotide polymorphisms (SNP in Mycobacterium leprae present on 115 Ziehl-Neelsen (Z-N-stained slides and in 51 skin biopsy samples derived from leprosy patients from Ceará (n = 23, Pernambuco (n = 41, Rio de Janeiro (n = 22 and Rondônia (RO (n = 78. All skin biopsies yielded SNP-based genotypes, while 48 of the samples (94.1% yielded complete VNTR genotypes. We evaluated two procedures for extracting M. leprae DNA from Z-N-stained slides: the first including Chelex and the other combining proteinase and sodium dodecyl sulfate. Of the 76 samples processed using the first procedure, 30.2% were positive for 16 or 15 VNTRs, whereas of the 39 samples processed using the second procedure, 28.2% yielded genotypes defined by at least 10 VNTRs. Combined VNTR and SNP analysis revealed large variability in genotypes, but a high prevalence of SNP genotype 4 in the Northeast Region of Brazil. Our observation of two samples from RO with an identical genotype and seven groups with similar genotypes, including four derived from residents of the same state or region, suggest a tendency to form groups according to the origin of the isolates. This study demonstrates the existence of geographically related M. leprae genotypes and that Z-N-stained slides are an alternative source for M. leprae genotyping.

  17. HLA class-II-restricted Mycobacterium leprae-reactive T-cell clones from leprosy patients established with a minimal requirement for autologous mononuclear cells

    NARCIS (Netherlands)

    Haanen, J. B.; Ottenhoff, T. H.; Voordouw, A.; Elferink, B. G.; Klatser, P. R.; Spits, H.; de Vries, R. R.

    1986-01-01

    This report describes an effective method for the cloning of Mycobacterium leprae-reactive T lymphocytes with Epstein-Barr-virus transformed autologous B cells as antigen-presenting cells. The two advantages of this method are that it drastically reduces the number of autologous peripheral blood

  18. qPCR-High resolution melt analysis for drug susceptibility testing of Mycobacterium leprae directly from clinical specimens of leprosy patients.

    Science.gov (United States)

    Araujo, Sergio; Goulart, Luiz Ricardo; Truman, Richard W; Goulart, Isabela Maria B; Vissa, Varalakshmi; Li, Wei; Matsuoka, Masanori; Suffys, Philip; Fontes, Amanda B; Rosa, Patricia S; Scollard, David M; Williams, Diana L

    2017-06-01

    Real-Time PCR-High Resolution Melting (qPCR-HRM) analysis has been recently described for rapid drug susceptibility testing (DST) of Mycobacterium leprae. The purpose of the current study was to further evaluate the validity, reliability, and accuracy of this assay for M. leprae DST in clinical specimens. The specificity and sensitivity for determining the presence and susceptibility of M. leprae to dapsone based on the folP1 drug resistance determining region (DRDR), rifampin (rpoB DRDR) and ofloxacin (gyrA DRDR) was evaluated using 211 clinical specimens from leprosy patients, including 156 multibacillary (MB) and 55 paucibacillary (PB) cases. When comparing the results of qPCR-HRM DST and PCR/direct DNA sequencing, 100% concordance was obtained. The effects of in-house phenol/chloroform extraction versus column-based DNA purification protocols, and that of storage and fixation protocols of specimens for qPCR-HRM DST, were also evaluated. qPCR-HRM results for all DRDR gene assays (folP1, rpoB, and gyrA) were obtained from both MB (154/156; 98.7%) and PB (35/55; 63.3%) patients. All PCR negative specimens were from patients with low numbers of bacilli enumerated by an M. leprae-specific qPCR. We observed that frozen and formalin-fixed paraffin embedded (FFPE) tissues or archival Fite's stained slides were suitable for HRM analysis. Among 20 mycobacterial and other skin bacterial species tested, only M. lepromatosis, highly related to M. leprae, generated amplicons in the qPCR-HRM DST assay for folP1 and rpoB DRDR targets. Both DNA purification protocols tested were efficient in recovering DNA suitable for HRM analysis. However, 3% of clinical specimens purified using the phenol/chloroform DNA purification protocol gave false drug resistant data. DNA obtained from freshly frozen (n = 172), formalin-fixed paraffin embedded (FFPE) tissues (n = 36) or archival Fite's stained slides (n = 3) were suitable for qPCR-HRM DST analysis. The HRM-based assay was also able to

  19. qPCR-High resolution melt analysis for drug susceptibility testing of Mycobacterium leprae directly from clinical specimens of leprosy patients.

    Directory of Open Access Journals (Sweden)

    Sergio Araujo

    2017-06-01

    Full Text Available Real-Time PCR-High Resolution Melting (qPCR-HRM analysis has been recently described for rapid drug susceptibility testing (DST of Mycobacterium leprae. The purpose of the current study was to further evaluate the validity, reliability, and accuracy of this assay for M. leprae DST in clinical specimens.The specificity and sensitivity for determining the presence and susceptibility of M. leprae to dapsone based on the folP1 drug resistance determining region (DRDR, rifampin (rpoB DRDR and ofloxacin (gyrA DRDR was evaluated using 211 clinical specimens from leprosy patients, including 156 multibacillary (MB and 55 paucibacillary (PB cases. When comparing the results of qPCR-HRM DST and PCR/direct DNA sequencing, 100% concordance was obtained. The effects of in-house phenol/chloroform extraction versus column-based DNA purification protocols, and that of storage and fixation protocols of specimens for qPCR-HRM DST, were also evaluated. qPCR-HRM results for all DRDR gene assays (folP1, rpoB, and gyrA were obtained from both MB (154/156; 98.7% and PB (35/55; 63.3% patients. All PCR negative specimens were from patients with low numbers of bacilli enumerated by an M. leprae-specific qPCR. We observed that frozen and formalin-fixed paraffin embedded (FFPE tissues or archival Fite's stained slides were suitable for HRM analysis. Among 20 mycobacterial and other skin bacterial species tested, only M. lepromatosis, highly related to M. leprae, generated amplicons in the qPCR-HRM DST assay for folP1 and rpoB DRDR targets. Both DNA purification protocols tested were efficient in recovering DNA suitable for HRM analysis. However, 3% of clinical specimens purified using the phenol/chloroform DNA purification protocol gave false drug resistant data. DNA obtained from freshly frozen (n = 172, formalin-fixed paraffin embedded (FFPE tissues (n = 36 or archival Fite's stained slides (n = 3 were suitable for qPCR-HRM DST analysis. The HRM-based assay was also able

  20. Factores de riesgo que influyen en la recaída de consumo de drogas lícitas e ilícitas en adolescentes atendidos en el Instituto sobre Alcoholismo y Farmacodependencia

    OpenAIRE

    Tatiana Blanco-Álvarez; Silvia Jiménez-Morales

    2015-01-01

    El objetivo de la presente investigación fue determinar los factores de riesgo asociados con las recaídas en el consumo de drogas en adolescentes que han recibido tratamiento en el IAFA. Metodología: Estudio cuantitativo, con alcance correlacional y transversal. Instrumentos: Entrevista de Recaídas para Usuarios de Conductas Adictivas, Inventario de Situaciones Precipitantes de Recaídas, Escala de Control Atencional, Inventario de Habilidades de Afrontamiento, Escala de Depresión, Ansiedad y ...

  1. Conocimientos de la población sobre lepra Knowledge of the population about leprosy

    Directory of Open Access Journals (Sweden)

    Isora Montenegro Valera

    2006-12-01

    Full Text Available Se realizó un estudio observacional descriptivo de tipo transversal para investigar el nivel de conocimientos, que sobre la lepra, tiene la población en el municipio de Limonar, durante el período de Marzo a Diciembre de 2002. Participaron en el estudio 395 pacientes mayores de 15 años, que fueron seleccionados de la población del municipio mediante un diseño multietápico que incluyó la estratificación y el conglomerado. Los datos fueron procesados en el sistema estadístico SPSS-10. Se utilizaron técnicas estadísticas como el Chi cuadrado para explorar la asociación significativa entre variables. Se obtuvo como resultado que existe desconocimiento por parte de la población acerca de la enfermedad, ya que solamente el 17,97 % de la población mostró conocimientos adecuados, y se encontró relación significativa entre este, el sexo femenino y la escolaridad. Sobre la base de los resultados se recomiendan las audiencias diana para una efectiva intervención educativa en la Atención Primaria de Salud en este municipio.An observational, descriptive and cross-sectional study was undertaken to investigate the level of knowledge of the population about leprosy in Limonar municipality from March to December, 2002. 395 patients over 15 participated in the study. They were selected from the population of the municipality by a multistage design that included stratification and cluster. The data were processed by the SPSS-10 statistical system. Statistical techniques as the Chi square test were used to explore the significant association among the variables. It was concluded that there is lack of knowledge about the disease, since only 17.97 % of the population showed an adequate knowledge. A significant relation was found among knowledge, the female sex and the educational level. According to these results, the target hearings are recommended for an effective educative intervention at the primary health care level in this municipality.

  2. Neuro-lepra: valor de la electromiografia Neuro-leprosy: electromyographic studies

    Directory of Open Access Journals (Sweden)

    Ernesto Herskovits

    1971-09-01

    Full Text Available Dada la frecuencia con que la lepra afecta al sistema nervioso, consideramos de interés realizar un estudio electromiográfico en zonas corporales clínicamente sanas. Hemos elegido para tal fin 14 enfermos que no tenían lesión sensitivo-motora clínicamente perceptible en el nervio cubital izquierdo. Hemos estudiado tambén un grupo de control de 5 enfermos con lesión evidente del mismo nervio. Se ha comprobado que de los 14 enfermos que aparentemente no tenían lesión del nervio cubital izquierdo, en 12 de ellos surgieron alteraciones electromiográficas que señalan la lesión del nervio, aunque en um grado menor que en el grupo de control. Este hecho nos hace pensar que la agresión que sufre el sistema nervioso periférico es de una extensión mayor que lo hace suponer la clínica, o que las lesiones anatómicas no retrogradan como nos lo sugiere el examen de los pacientes.Considering the frequency of the peripheral nervous system envolvement in leprosy 14 patients without clinical signs indicating impairment of the left ulnar nerve were submitted to electromyographic studies. All were chronic cases in which the disease had an evolution of three years for the most recent one, the longest during thirty one years. All patients were under leprosy treatment: nine had lepromatous leprosy, four had tuberculoid form, one had a dimorfous form. At the same time, as a control group, were studied 5 patients presenting clinical signis of injury of the left ulnar nerve. An electromiograph DISA with 3 channels, a Multistin estimulator and concentric electrodes were employed. In all the 19 cases the espontaneous activity, the type of recruiting reaction and the conduction velocity were analysed. Results were synthetized in Tables 1 and 2. The finding of electromyographic abnormalities in clinically healthy territores of 12/14 patients examined lead to the conclusion that in leprosy the agression to the peripheral nervous system is more extensive than

  3. How long should older people take antidepressants to prevent relapse? Por quanto tempo os idosos devem tomar antidepressivos para evitar recaídas?

    Directory of Open Access Journals (Sweden)

    Jon Spear

    2002-04-01

    Full Text Available Patients with depressive disorder have a high risk of relapse after recovery from a depressive episode. Can the relapse of depressive disorder be prevented or delayed for older adults? This paper reviews the evidence from randomised clinical trials and open label trials of the effectiveness of maintenance antidepressant therapy for older adults with depressive disorder. It also examines the evidence for the effectiveness of psychosocial and psychotherapeutic interventions. The paper concludes with recommendations for clinical practice and future research.Pacientes com transtorno depressivo apresentam alto risco de recorrência e recaída. É possível prevenir a recaída ou a recorrência do episódio depressivo ou retardá-lo em fases tardias da vida? Este artigo revisa ensaios clínicos aleatorizados e não-aleatorizados com o objetivo de estabelecer se o tratamento antidepressivo de manutenção reduz o risco de recaída e recorrência de depressão em idosos. O artigo também examina a evidência atualmente disponível sobre a eficácia das intervenções psicossociais e psicoterapêuticas. O artigo conclui com recomendações para a prática clínica e pesquisas futuras.

  4. Role of RecA protein in untargeted UV mutagenesis of bacteriophage lambda: evidence for the requirement for the dinB gene

    International Nuclear Information System (INIS)

    Brotcorne-Lannoye, A.; Maenhaut-Michel, G.

    1986-01-01

    Untargeted UV mutagenesis of bacteriophage lambda--i.e., the increased recovery of lambda mutants when unirradiated lambda infects UV-irradiated Escherichia coli--is thought to be mediated by a transient decrease in DNA replication fidelity, generating mutations in the newly synthesized strands. Using the bacteriophage lambda cI857----lambda c mutation system, we provide evidence that the RecA protein, shown previously to be required for this mutagenic pathway, is no longer needed when the LexA protein is inactivated by mutation. We suggest that the error-prone DNA replication responsible for UV-induced untargeted mutagenesis is turned on by the presence of replication-blocking lesions in the host cell DNA and that the RecA protein is required only to derepress the relevant din gene(s). This is in contrast to mutagenesis of irradiated bacteria or irradiated phage lambda, in which activated RecA protein has a second role in mutagenesis in addition to the cleavage of the LexA protein. Among the tested din genes, the dinB gene product (in addition to the uvrA and uvrB gene products) was found to be required for untargeted mutagenesis of bacteriophage lambda. To our knowledge, a phenotype associated with the dinB gene has not been reported previously

  5. Tratamento eclético da lepra: 20 anos de experiência

    Directory of Open Access Journals (Sweden)

    H. C. de Souza-Araújo

    1949-06-01

    a false measure of the severity of the disease; others remained very high notwithstanding the great improvement of the disease. All patients with more than 12 months treatment became practically symptom free. Lepra reaction amongst them was rare and always started by embolic rash, being controlled by destruction of such skin lesions by galvanocauterisation. In a few cases the lepromata infiltrated with "Subintrol" (a 3 p. c. special chaulmoogra soap prepared by Dr. ASTROGILDO MACHADO were completely destroyd and never relapsed. In October 1946 Dr. ERNEST MUIR saw here a few cases treated by the author's method and suggested the combination of Diasone with galvanocauterisation which is being done now with satifactory results. The second part of this paper, reporting many leprosy cases treated by the so-called ecclectical method, which are symptom free and negativated since five to ten years, will be published as soon as the sulfone-therapy be summarised in some reliable scientific report to be compared with.

  6. Prophage induction and differential RecA and UmuDAb transcriptome regulation in the DNA damage responses of Acinetobacter baumannii and Acinetobacter baylyi.

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    Janelle M Hare

    Full Text Available The SOS response to DNA damage that induces up to 10% of the prokaryotic genome requires RecA action to relieve LexA transcriptional repression. In Acinetobacter species, which lack LexA, the error-prone polymerase accessory UmuDAb is instead required for ddrR induction after DNA damage, suggesting it might be a LexA analog. RNA-Seq experiments defined the DNA damage transcriptome (mitomycin C-induced of wild type, recA and umuDAb mutant strains of both A. baylyi ADP1 and A. baumannii ATCC 17978. Of the typical SOS response genes, few were differentially regulated in these species; many were repressed or absent. A striking 38.4% of all ADP1 genes, and 11.4% of all 17978 genes, were repressed under these conditions. In A. baylyi ADP1, 66 genes (2.0% of the genome, including a CRISPR/Cas system, were DNA damage-induced, and belonged to four regulons defined by differential use of recA and umuDAb. In A. baumannii ATCC 17978, however, induction of 99% of the 152 mitomycin C-induced genes depended on recA, and only 28 of these genes required umuDAb for their induction. 90% of the induced A. baumannii genes were clustered in three prophage regions, and bacteriophage particles were observed after mitomycin C treatment. These prophages encoded esvI, esvK1, and esvK2, ethanol-stimulated virulence genes previously identified in a Caenorhabditis elegans model, as well as error-prone polymerase alleles. The induction of all 17978 error-prone polymerase alleles, whether prophage-encoded or not, was recA dependent, but only these DNA polymerase V-related genes were de-repressed in the umuDAb mutant in the absence of DNA damage. These results suggest that both species possess a robust and complex DNA damage response involving both recA-dependent and recA-independent regulons, and further demonstrates that although umuDAb has a specialized role in repressing error-prone polymerases, additional regulators likely participate in these species' transcriptional

  7. New Insights into the Geographic Distribution of Mycobacterium leprae SNP Genotypes Determined for Isolates from Leprosy Cases Diagnosed in Metropolitan France and French Territories.

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    Florence Reibel

    Full Text Available Between 20 and 30 bacteriologically confirmed cases of leprosy are diagnosed each year at the French National Reference Center for mycobacteria. Patients are mainly immigrants from various endemic countries or living in French overseas territories. We aimed at expanding data regarding the geographical distribution of the SNP genotypes of the M. leprae isolates from these patients.Skin biopsies were obtained from 71 leprosy patients diagnosed between January 2009 and December 2013. Data regarding age, sex and place of birth and residence were also collected. Diagnosis of leprosy was confirmed by microscopic detection of acid-fast bacilli and/or amplification by PCR of the M. leprae-specific RLEP region. Single nucleotide polymorphisms (SNP, present in the M. leprae genome at positions 14 676, 1 642 875 and 2 935 685, were determined with an efficiency of 94% (67/71. Almost all patients were from countries other than France where leprosy is still prevalent (n = 31 or from French overseas territories (n = 36 where leprosy is not totally eradicated, while only a minority (n = 4 was born in metropolitan France but have lived in other countries. SNP type 1 was predominant (n = 33, followed by type 3 (n = 17, type 4 (n = 11 and type 2 (n = 6. SNP types were concordant with those previously reported as prevalent in the patients' countries of birth. SNP types found in patients born in countries other than France (Comoros, Haiti, Benin, Congo, Sri Lanka and French overseas territories (French Polynesia, Mayotte and La Réunion not covered by previous work correlated well with geographical location and history of human settlements.The phylogenic analysis of M. leprae strains isolated in France strongly suggests that French leprosy cases are caused by SNP types that are (a concordant with the geographic origin or residence of the patients (non-French countries, French overseas territories, metropolitan France or (b more likely random in regions where

  8. New Insights into the Geographic Distribution of Mycobacterium leprae SNP Genotypes Determined for Isolates from Leprosy Cases Diagnosed in Metropolitan France and French Territories.

    Science.gov (United States)

    Reibel, Florence; Chauffour, Aurélie; Brossier, Florence; Jarlier, Vincent; Cambau, Emmanuelle; Aubry, Alexandra

    2015-01-01

    Between 20 and 30 bacteriologically confirmed cases of leprosy are diagnosed each year at the French National Reference Center for mycobacteria. Patients are mainly immigrants from various endemic countries or living in French overseas territories. We aimed at expanding data regarding the geographical distribution of the SNP genotypes of the M. leprae isolates from these patients. Skin biopsies were obtained from 71 leprosy patients diagnosed between January 2009 and December 2013. Data regarding age, sex and place of birth and residence were also collected. Diagnosis of leprosy was confirmed by microscopic detection of acid-fast bacilli and/or amplification by PCR of the M. leprae-specific RLEP region. Single nucleotide polymorphisms (SNP), present in the M. leprae genome at positions 14 676, 1 642 875 and 2 935 685, were determined with an efficiency of 94% (67/71). Almost all patients were from countries other than France where leprosy is still prevalent (n = 31) or from French overseas territories (n = 36) where leprosy is not totally eradicated, while only a minority (n = 4) was born in metropolitan France but have lived in other countries. SNP type 1 was predominant (n = 33), followed by type 3 (n = 17), type 4 (n = 11) and type 2 (n = 6). SNP types were concordant with those previously reported as prevalent in the patients' countries of birth. SNP types found in patients born in countries other than France (Comoros, Haiti, Benin, Congo, Sri Lanka) and French overseas territories (French Polynesia, Mayotte and La Réunion) not covered by previous work correlated well with geographical location and history of human settlements. The phylogenic analysis of M. leprae strains isolated in France strongly suggests that French leprosy cases are caused by SNP types that are (a) concordant with the geographic origin or residence of the patients (non-French countries, French overseas territories, metropolitan France) or (b) more likely random in regions where diverse

  9. DNA degradation, UV sensitivity and SOS-mediated mutagenesis in strains of Escherichia coli deficient in single-strand DNA binding protein: Effects of mutations and treatments that alter levels of exonuclease V or RecA protein

    International Nuclear Information System (INIS)

    Lieberman, H.B.; Witkin, E.M.

    1983-01-01

    Certain strains suppress the temperature-sensitivity caused by ssb-1, which encodes a mutant ssDNA binding protein (SSB). At 42 0 C, such strains are extremely UV-sensitive, degrade their DNA extensively after UV irradiation, and are defficient in UV mutability and UV induction of recA protein synthesis. We transduced recC22, which eliminates Exonuclease V activity, and recAo281, which causes operator-constitutive synthesis of recA protein, into such an ssb-1 strain. Both double mutants degraded their DNA extensively at 42 0 C after UV irradiation, and both were even more UV-sensitive than the ssb-1 single mutant. We conclude that one or more nucleases other than Exonuclease V degrades DNA in the ssb recC strain, and that recA protein, even if synthesized copiously, can function efficiently in recombinational DNA repair and in control of post-UV DNA degradation only if normal SSB is also present. Pretreatment with nalidixic acid at 30 0 C restored normal UV mutability at 42 0 C, but did not increase UV resistance, in an ssb-1 strain. Another ssb allele, ssb-113, which blocks SOS induction at 30 0 C, increases spontaneous mutability more than tenfold. The ssb-113 allele was transduced into the SOS-constitutive recA730 strain SC30. This double mutant expressed the same elevated spontaneous and UV-induced mutability at 30 0 C as the ssb + recA730 strain, and was three times more UV-resistant than its ssb-113 recA + parent. We conclude that ssb-1 at 42 0 C and ssb-113 at 30 0 C block UV-induced activation of recA protease, but that neither allele interferes with subsequent steps in SOS-mediated mutagenesis. (orig.)

  10. RecET driven chromosomal gene targeting to generate a RecA deficient Escherichia coli strain for Cre mediated production of minicircle DNA

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    Coutelle Charles

    2006-03-01

    Full Text Available Abstract Background Minicircle DNA is the non-replicating product of intramolecular site-specific recombination within a bacterial minicircle producer plasmid. Minicircle DNA can be engineered to contain predominantly human sequences which have a low content of CpG dinucleotides and thus reduced immunotoxicity for humans, whilst the immunogenic bacterial origin and antibiotic resistance marker gene sequences are entirely removed by site-specific recombination. This property makes minicircle DNA an excellent vector for non-viral gene therapy. Large-scale production of minicircle DNA requires a bacterial strain expressing tightly controlled site-specific recombinase, such as Cre recombinase. As recombinant plasmids tend to be more stable in RecA-deficient strains, we aimed to construct a recA- bacterial strain for generation of minicircle vector DNA with less chance of unwanted deletions. Results We describe here the construction of the RecA-deficient minicircle DNA producer Escherichia coli HB101Cre with a chromosomally located Cre recombinase gene under the tight control of the araC regulon. The Cre gene expression cassette was inserted into the chromosomal lacZ gene by creating transient homologous recombination proficiency in the recA- strain HB101 using plasmid-born recET genes and homology-mediated chromosomal "pop-in, pop-out" of the plasmid pBAD75Cre containing the Cre gene and a temperature sensitive replication origin. Favourably for the Cre gene placement, at the "pop-out" step, the observed frequency of RecET-led recombination between the proximal regions of homology was 10 times higher than between the distal regions. Using the minicircle producing plasmid pFIXluc containing mutant loxP66 and loxP71 sites, we isolated pure minicircle DNA from the obtained recA- producer strain HB101Cre. The minicircle DNA preparation consisted of monomeric and, unexpectedly, also multimeric minicircle DNA forms, all containing the hybrid loxP66

  11. Evolución de la endemia de la lepra en Brasil Time trends of Hansen's disease in Brasil

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    Maria da Conceição Cavalcanti Magalhães

    2005-12-01

    Full Text Available La presente investigación se propone abordar la distribución territorial de la lepra en Brasil con el objetivo de identificar las regularidades de su diferenciación espacial y aportar conocimientos que permitan progresar en la comprensión de los procesos de su producción social. Fueron calculadas las tasas anuales de detección de casos, brutos y específicos, para las regiones y estados del país. La evolución espacio-temporal fue realizada mediante la construcción de series históricas. Se expone como substrato de los análisis una síntese sobre la distribución histórico-geográfica de la endemia en Brasil. La serie histórica del período 1960 - 2002, permite comparar las tasas de detección de las regiones, en la primera década, con las tasas más bajas, a excepción de la región Norte, que se destaca com la mayor detección. Todas las regiones elevan la detección a partir de la década del 70, siendo la Sul la de detección más baja. Las regiones Norte y Centro Oeste, mantienen las más altas detecciones de casos y los mayores incrementos, lo cual evidencia la desigual evolución regional de la endemia en el país. La permanencia histórica de elevada incidencia de la lepra en la región Norte, en el estado de Maranhão y Pernambuco en la región Nordeste, del estado de Paraná en la región Sul, así como relativamente bajas incidencias en el resto de esta última región, sugieren la existencia de contextos geográficos de diferente vulnerabilidad a la producción social de la lepra, y de nuevos componentes en la organización espacial en algunos territorios.The present investigation intends to study the territorial distribution of leprosy in Brazil in order to identify the regularities of its differentiation, and to contribute to the knowledge that allows progress in the understanding of the processes of its social production. Time trends of global and specific leprosy detection rates were calculated for regions and states

  12. Recaída neurológica en perros con linfoma tratados con quimioterapia: revisión de 13 casos

    OpenAIRE

    Martínez de Merlo, E. M.

    2011-01-01

    Se realiza un estudio retrospectivo de 13 casos de perros tratados con quimioterapia por linfoma que sufren recaída con afectación del sistema nervioso central o periférico, con el fin de describir los hallazgos encontrados y respuesta al tratamiento de los mismos. No se ha encontrado correlación estadística entre sexo, edad, estadio clínico o forma de presentación de la enfermedad neoplásica con la aparición de signos neurológicos. Sin embargo, se ha demostrado una predisposición racial a de...

  13. The approaches to mathematical modeling of recA, umuD genes expression in bacteria Escherichia coli after UV-irradiation

    International Nuclear Information System (INIS)

    Belov, O.V.

    2006-01-01

    The modern data of recA, umuD genes expression of the system of SOS-repair at classical object of radiation genetic researches - bacteria Escherichia coli, after ultraviolet irradiation are presented. Essentially a new method of analysis of SOS-genes expression is considered. It was shown that using this method it is possible to determine the character of induction of some SOS-genes more precisely. The possible approach to the mathematical description of SOS-response of cells by construction of the system of the differential equations is presented

  14. Exploring the Molecular Basis for Selective Binding of Homoserine Dehydrogenase from Mycobacterium leprae TN toward Inhibitors: A Virtual Screening Study

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    Dongling Zhan

    2014-01-01

    Full Text Available Homoserine dehydrogenase (HSD from Mycobacterium leprae TN is an antifungal target for antifungal properties including efficacy against the human pathogen. The 3D structure of HSD has been firmly established by homology modeling methods. Using the template, homoserine dehydrogenase from Thiobacillus denitrificans (PDB Id 3MTJ, a sequence identity of 40% was found and molecular dynamics simulation was used to optimize a reliable structure. The substrate and co-factor-binding regions in HSD were identified. In order to determine the important residues of the substrate (l-aspartate semialdehyde (l-ASA binding, the ASA was docked to the protein; Thr163, Asp198, and Glu192 may be important because they form a hydrogen bond with HSD through AutoDock 4.2 software. neuraminidaseAfter use of a virtual screening technique of HSD, the four top-scoring docking hits all seemed to cation–π ion pair with the key recognition residue Lys107, and Lys207. These ligands therefore seemed to be new chemotypes for HSD. Our results may be helpful for further experimental investigations.

  15. Exploring the molecular basis for selective binding of homoserine dehydrogenase from Mycobacterium leprae TN toward inhibitors: a virtual screening study.

    Science.gov (United States)

    Zhan, Dongling; Wang, Dongmei; Min, Weihong; Han, Weiwei

    2014-01-24

    Homoserine dehydrogenase (HSD) from Mycobacterium leprae TN is an antifungal target for antifungal properties including efficacy against the human pathogen. The 3D structure of HSD has been firmly established by homology modeling methods. Using the template, homoserine dehydrogenase from Thiobacillus denitrificans (PDB Id 3MTJ), a sequence identity of 40% was found and molecular dynamics simulation was used to optimize a reliable structure. The substrate and co-factor-binding regions in HSD were identified. In order to determine the important residues of the substrate (L-aspartate semialdehyde (L-ASA)) binding, the ASA was docked to the protein; Thr163, Asp198, and Glu192 may be important because they form a hydrogen bond with HSD through AutoDock 4.2 software. neuraminidaseAfter use of a virtual screening technique of HSD, the four top-scoring docking hits all seemed to cation-π ion pair with the key recognition residue Lys107, and Lys207. These ligands therefore seemed to be new chemotypes for HSD. Our results may be helpful for further experimental investigations.

  16. SOS-like induction in Bacillus subtilis: induction of the RecA protein analog and a damage-inducible operon by DNA damage in Rec+ and DNA repair-deficient strains

    International Nuclear Information System (INIS)

    Lovett, C.M. Jr.; Love, P.E.; Yasbin, R.E.; Roberts, J.W.

    1988-01-01

    We quantitated the induction of the Bacillus subtilis Rec protein (the analog of Escherichia coli RecA protein) and the B. subtilis din-22 operon (representative of a set of DNA damage-inducible operons in B. subtilis) following DNA damage in Rec+ and DNA repair-deficient strains. After exposure to mitomycin C or UV irradiation, each of four distinct rec (recA1, recB2, recE4, and recM13) mutations reduced to the same extent the rates of both Rec protein induction (determined by densitometric scanning of immunoblot transfers) and din-22 operon induction (determined by assaying beta-galactosidase activity in din-22::Tn917-lacZ fusion strains). The induction deficiencies in recA1 and recE4 strains were partially complemented by the E. coli RecA protein, which was expressed on a plasmid in B. subtilis; the E. coli RecA protein had no effect on either induction event in Rec+, recB2, or recM13 strains. These results suggest that (i) the expression of both the B. subtilis Rec protein and the din-22 operon share a common regulatory component, (ii) the recA1 and recE4 mutations affect the regulation and/or activity of the B. subtilis Rec protein, and (iii) an SOS regulatory system like the E. coli system is highly conserved in B. subtilis. We also showed that the basal level of B. subtilis Rec protein is about 4,500 molecules per cell and that maximum induction by DNA damage causes an approximately fivefold increase in the rate of Rec protein accumulation

  17. Over a Decade of recA and tly Gene Sequence Typing of the Skin Bacterium Propionibacterium acnes: What Have We Learnt?

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    Andrew McDowell

    2017-12-01

    Full Text Available The Gram-positive, anaerobic bacterium Propionibacterium acnes forms part of the normal microbiota on human skin and mucosal surfaces. While normally associated with skin health, P. acnes is also an opportunistic pathogen linked with a range of human infections and clinical conditions. Over the last decade, our knowledge of the intraspecies phylogenetics and taxonomy of this bacterium has increased tremendously due to the introduction of DNA typing schemes based on single and multiple gene loci, as well as whole genomes. Furthermore, this work has led to the identification of specific lineages associated with skin health and human disease. In this review we will look back at the introduction of DNA sequence typing of P. acnes based on recA and tly loci, and then describe how these methods provided a basic understanding of the population genetic structure of the bacterium, and even helped characterize the grapevine-associated lineage of P. acnes, known as P. acnes type Zappe, which appears to have undergone a host switch from humans-to-plants. Particular limitations of recA and tly sequence typing will also be presented, as well as a detailed discussion of more recent, higher resolution, DNA-based methods to type P. acnes and investigate its evolutionary history in greater detail.

  18. Current strategy for leprosy control in Brazil: time to pursue alternative preventive strategies? Estrategia actual para el control de la lepra en Brasil: ¿es hora de investigar otras estrategias de prevención?

    OpenAIRE

    Sérgio S. Cunha; Laura C. Rodrigues; Nádia Cristina Duppre

    2004-01-01

    La estrategia actual para el control de la lepra en Brasil se basa en dos actividades principales: la detección precoz de casos y el tratamiento de casos con farmacoterapia combinada. Además de dichas medidas, se realizan esfuerzos complementarios para identificar los contactos domésticos para el diagnóstico precoz y la vacunación con el bacilo de Calmette-Guérin (BCG). Sin embargo, la eficacia de estas acciones a la hora de reducir la incidencia de la lepra es aún discutible. Esto genera dud...

  19. Estudio de resistencia a la rifampicina y la dapsona en tres pacientes con recurrencia de lepra Study of rifampin and dapsone resistance in three patients with recurring leprosy

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    Elkin Hernández

    2008-02-01

    Full Text Available OBJETIVO: Detectar la presencia de cepas de Mycobacterium leprae resistentes a la rifampicina y la dapsona en tres pacientes con recurrencia de lepra y sospecha clínica de resistencia antimicrobiana, mediante la aplicación de técnicas moleculares. MÉTODOS: Se realizó un estudio descriptivo retrospectivo en tres pacientes multibacilares del Sanatorio de Agua de Dios, Cundinamarca, Colombia, que habían presentado recidivas de lepra documentadas por su historia clínica, baciloscopia y biopsia. Se obtuvieron biopsias de lesiones cutáneas que se procesaron para la extracción y purificación del ADN bacilar. Se amplificaron regiones de los genes rpoB y folP1 asociadas con la resistencia antimicrobiana, mediante la reacción en cadena de la polimerasa "touch-down" y se secuenciaron los productos amplificados mediante el método de Sanger. RESULTADOS: Se detectó una mutación puntual en el nucleótido 1367 del gen rpoB en dos de las muestras estudiadas. No se encontró la mutación estudiada en el gen folP1 en ninguno de los tres pacientes. CONCLUSIONES: La mutación identificada demostró la presencia de bacilos de M. leprae resistentes a la rifampicina en dos de los tres pacientes estudiados con recurrencia de la enfermedad. No se detectó la mutación indicadora de resistencia a la dapsona en ninguno de los tres pacientes.OBJECTIVE: To detect the presence of rifampin- and dapsone-resistant strains of Mycobacterium leprae in three patients with recurring leprosy and clinically-suspected antimicrobial resistance through molecular techniques. METHODS: A retrospective, descriptive study was conducted of three multibacillary patients at the "Agua de Dios" Sanitarium in Cundinamarca, Colombia, that presented leprosy relapses that were documented by medical history, bacilloscopy, and biopsy. Biopsies were taken of the skin lesions and the bacteria were subject to DNA extraction and purification. Regions of the rpoB and folP1 genes associated with

  20. Hanseníase: a realidade do ser adolescente Lepra: la realidad para el adolescente Leprosy: the reality for the adolescent

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    Keila Maria de Azevedo Ponte

    2005-06-01

    Full Text Available A hanseníase é uma doença contagiosa, estigmatizante, de grande potencial incapacitante, e os adolescentes por estarem numa fase de mudanças e de adaptação, esta doença pode interferir na construção de sua vida. O estudo objetiva caracterizar os adolescentes portadores de hanseníase segundo aspectos sócio-demograficos; realizar análise epidemiológico-operacional da hanseníase; verificar o conhecimento dos adolescentes sobre a hanseníase e as reações manifestadas pelos mesmos após sua descoberta; Identificar as mudanças ocorridas após a doença na vida do adolescente e as dificuldades vivenciadas pelo mesmo após a da doença. Trata-se de uma pesquisa exploratória e descritiva, constituída de 31 adolescentes portadores de hanseníase, assistido pela Estratégia Saúde da Família no Município de Sobral- Ceará. Os resultados sinalizam a necessidade de uma assistência integral e continuada ao adolescente portador de hanseníase, evitando que essa doença provoque mudanças significativas em sua vida, dificultando na construção de sua nova identidade.La lepra es una enfermedad contagiosa, de un gran potencial incapacitante y los adolescentes por estaren viviendo en una fase de cambios e adaptación esta enfermedad puede interferir en la construcción de sus vidas. Esta investigación apunta caracterizar los adolescentes portadores de lepra según aspectos sociales-demográficos; hacer un análisis epidémico-operacional de la enfermedad lepra; verificar el conocimiento de los adolescentes sobre la lepra y las relaciones manifestadas por los mismos después de su descubierta; identificar los cambios ocurridos en la vida del adolescente después de la descubierta de la enfermedad. Tratase de una investigación exploratoria y descriptiva donde participarón 31 adolescentes portadores de lepra ayudados por la Estrategia Salud de la Familia en el municipio de Sobral - Ceará. Los resultados señalan la necesidad de una

  1. Possibilidades de transmissão e vias de inoculação da lepra murina em ratos e outros animais

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    Herminio Linhares

    1943-06-01

    Full Text Available 1 O A. revê as vias de infecção naturais e os processos de inoculações empregados em ratos, no estudo da lepra murina. 2 Na natureza, o contacto prolongado de animal sadio com doente e a infecção por via gástrica devem ser os modos comuns de contaminação. 3 Foram encontrados dentro do Polyplax spinulosa (Burmeister capturados em ratos leprosos, bacilos ácido álcool resistentes. Tentativas de cultura com êste material, foram infrutíferas. 4 O A. infectou ratos colocando no estômago, por meio de sondas de vidro, material leproso. Em cinco animais, todos se infectaram. 5 Por via subcutânea e por via intraperitoneal, a infecção se processa em quase 100% dos casos. 6 Foi possível infectar gambás (Didelphis aurita com lepra murina. Êsses animais provavelmente são mais suscetíveis à lepra dos ratos que à humana. 7 Conseguiu-se infectar pinto por inoculação de emulsão de lepra murina no músculo do peito, por via intraperitoneal e por via gástrica. 8 Pombos também se infectaram após inoculação no músculo do peito e por via venosa.1 The A. reviews the routes of natural transmission of rat leprosy and the experimentally induced disease. 2 The infection in the natural disease must be made by contact with an infected rat or through the gastro-intestinal route by eating infected tissue. 3 They were found acid-fast bacilli in lice (Polyplax spinulosa caught on rats dying of leprosy; but it was impossible to obtain cultures in Löwenstein medium, from these lice. 4 Rat leprosy emulsion introduced into the stomach, may infect rats. Five rats fed with infected material became infected. 5 After subcutaneous or intraperitoneal inoculation there were obtained infection in almost 100% of cases. 6 It was possible to infect Didelphis aurita after inoculation of infected rat material. These animals most likely are more susceptible to rat leprosy than to human leprosy. 7 It was possible to infect chicks by inoculation in chest muscle

  2. Discovery of a potential lead compound for treating leprosy with dapsone resistance mutation in M. leprae folP1.

    Science.gov (United States)

    Nisha, J; Ramanathan, K; Nawaz Khan, F; Dhanasekaran, D; Shanthi, V

    2016-06-21

    Dapsone resistance is a serious impediment to the implementation of the present leprosy control strategies. In the recent past, many studies have been undertaken to address the antibiotic activity and binding pattern of dapsone against both native and mutant (Pro55Leu) folP1. Yet, there is no well-developed structural basis for understanding drug action and there is dire need for new antibacterial therapies. In the present study, molecular simulation techniques were employed alongside experimental strategies to address and overcome the mechanism of dapsone resistance. In essence, we report the identification of small molecule compounds to effectively and specifically inhibit the growth of M. leprae through targeting dihydropteroate synthase, encoded by folP1 which is involved in folic acid synthesis. Initially, ADME and toxicity studies were employed to screen the lead compounds, using dapsone as standard drug. Subsequently, molecular docking was employed to understand the binding efficiency of dapsone and its lead compounds against folP1. Further, the activity of the screened lead molecule was studied by means of molecular dynamics simulation techniques. Furthermore, we synthesized 4-(2-fluorophenylsulfonyl)benzenamine, using (2-fluorophenyl)boronic acid and 4-aminobenzenesulfonyl chloride, and the compound structure was confirmed by (1)H NMR and (13)C NMR spectroscopic techniques. Most importantly, the antibacterial activity of the compound was also examined and compared against dapsone. Overall, the result from our analysis suggested that CID21480113 (4-(2-fluorophenylsulfonyl)benzenamine) could be developed into a promising lead compound and could be effective in treating dapsone resistant leprosy cases.

  3. Drug targeted virtual screening and molecular dynamics of LipU protein of Mycobacterium tuberculosis and Mycobacterium leprae.

    Science.gov (United States)

    Kaur, Gurkamaljit; Pandey, Bharati; Kumar, Arbind; Garewal, Naina; Grover, Abhinav; Kaur, Jagdeep

    2018-03-20

    The lipolytic protein LipU was conserved in mycobacterium sp. including M. tuberculosis (MTB LipU) and M. leprae (MLP LipU). The MTB LipU was identified in extracellular fraction and was reported to be essential for the survival of mycobacterium. Therefore to address the problem of drug resistance in pathogen, LipU was selected as a drug target and the viability of finding out some FDA approved drugs as LipU inhibitors in both the cases was explored. Three dimensional (3D) model structures of MTB LipU and MLP LipU were generated and stabilized through molecular dynamics (MD). FDA approved drugs were screened against these proteins. The result showed that the top-scoring compounds for MTB LipU were Diosmin, Acarbose and Ouabain with the Glide XP score of -12.8, -11.9 and-11.7 kcal/mol respectively, whereas for MLP LipU protein, Digoxin (-9.2 kcal/mol), Indinavir (-8.2 kcal/mol) and Travoprost (-8.2 kcal/mol) showed highest affinity. These drugs remained bound in the active site pocket of MTB LipU and MLP LipU structure and interaction grew stronger after dynamics. RMSD, RMSF and Rg were found to be persistent throughout the simulation period. Hydrogen bonds along with large number of hydrophobic interactions stabilized the complex structures. Binding free energies obtained through Prime/MM-GBSA were found in the significant range from -63.85 kcal/mol to -34.57 kcal/mol for MTB LipU and -71.33 kcal/mol to -23.91 kcal/mol for MLP LipU. The report suggested high probability of these drugs to demolish the LipU activity and could be probable drug candidates to combat TB and leprosy disease.

  4. Crystal structure of the left-handed archaeal RadA helical filament: identification of a functional motif for controlling quaternary structures and enzymatic functions of RecA family proteins

    Science.gov (United States)

    Chen, Li-Tzu; Ko, Tzu-Ping; Chang, Yuan-Chih; Lin, Kuei-An; Chang, Chia-Seng; Wang, Andrew H.-J.; Wang, Ting-Fang

    2007-01-01

    The RecA family of proteins mediates homologous recombination, an evolutionarily conserved pathway that maintains genomic stability by protecting against DNA double strand breaks. RecA proteins are thought to facilitate DNA strand exchange reactions as closed-rings or as right-handed helical filaments. Here, we report the crystal structure of a left-handed Sulfolobus solfataricus RadA helical filament. Each protomer in this left-handed filament is linked to its neighbour via interactions of a β-strand polymerization motif with the neighbouring ATPase domain. Immediately following the polymerization motif, we identified an evolutionarily conserved hinge region (a subunit rotation motif) in which a 360° clockwise axial rotation accompanies stepwise structural transitions from a closed ring to the AMP–PNP right-handed filament, then to an overwound right-handed filament and finally to the left-handed filament. Additional structural and functional analyses of wild-type and mutant proteins confirmed that the subunit rotation motif is crucial for enzymatic functions of RecA family proteins. These observations support the hypothesis that RecA family protein filaments may function as rotary motors. PMID:17329376

  5. Factores de riesgo que influyen en la recaída de consumo de drogas lícitas e ilícitas en adolescentes atendidos en el Instituto sobre Alcoholismo y Farmacodependencia

    Directory of Open Access Journals (Sweden)

    Tatiana Blanco-Álvarez

    2015-12-01

    Full Text Available El objetivo de la presente investigación fue determinar los factores de riesgo asociados con las recaídas en el consumo de drogas en adolescentes que han recibido tratamiento en el IAFA. Metodología: Estudio cuantitativo, con alcance correlacional y transversal. Instrumentos: Entrevista de Recaídas para Usuarios de Conductas Adictivas, Inventario de Situaciones Precipitantes de Recaídas, Escala de Control Atencional, Inventario de Habilidades de Afrontamiento, Escala de Depresión, Ansiedad y Estrés, Escala Multidimensional de Apoyo Social Recibido, Escala de Dificultades en la Regulación Emocional, y Cuestionario Breve de Confianza Situacional. Muestra: 107 adolescentes que reciben atención ambulatoria en el Centro de Menores del Instituto de Alcoholismo y Farmacodependencia. Conclusiones: las variables intrapersonales como regulación emocional (conductas dirigidas a metas, control atencional (atención focalizada, estados emocionales negativos y búsqueda de sensaciones positivas son factores de riesgo determinantes en las recaídas de consumo de drogas.

  6. Evaluation of Three Mycobacterium leprae Monoclonal Antibodies in Mucus and Lymph Samples from Ziehl-Neelsen Stain Negative Leprosy Patients and their Household Contacts in an Indian Community

    Directory of Open Access Journals (Sweden)

    Nora Cardona-Castro

    1998-07-01

    Full Text Available Mucus and lymph smears collected from leprosy patients (9 and their household contacts (44 in the Caño Mochuelo Indian Reservation, Casanare, Colombia, were examined with monoclonal antibodies (MoAb against Mycobacterium leprae. The individuals studied were: 5 borderline leprosy (BB patients, 4 with a lepromatous leprosy (LL, all of whom were undergoing epidemiological surveillance after treatment and 44 household contacts: 21 of the LL and 23 contacts of the BB patients. The MoAb were reactive with the following M. leprae antigens: 65 kd heat shock protein, A6; soluble antigen G7 and complete antigen, E11. All the samples were tested with each of the MoAb using the avidin-biotin-peroxidase technique and 3,3 diaminobenzidine as chromogen. The patients and household contacts studied were all recorded as Ziehl-Neelsen stain negative. The MoAb which showed optimal reaction was G7, this MoAb permited good visualization of the bacilli. Five patients with BB diagnosis and one with LL were positive for G7; of the BB patients' household contacts, 9 were positive for G7; 7 of the LL patients' household contacts were positive for the same MoAb. MoAb G7 allowed the detection of bacillar Mycobacterium spp. compatible structures in both patients and household contacts. G7 permited the visualization of the complete bacillus and could be used for early diagnosis and follow-up of the disease in patients.

  7. Lepra y estados reaccionales. A propósito de un caso y revisión bibliográfica

    Directory of Open Access Journals (Sweden)

    Petty Bonivento

    2013-10-01

    Full Text Available  Resumen La lepra es una infección bacteriana crónica causada por el Mycobacterium leprae, bacilo ácido-alcohol resistente obligado a vivir en el espacio intracelular preferentemente en las células de Schwann y macrófagos, que afecta piel, nervios y ojos principalmente. La existencia de la patología se conoce desde la antigüedad, pero aun continúa siendo un grave problema de salud pública a nivel mundial sobre todo en áreas endémicas. Su espectro clínico está dado por la respuesta que genera el sistema inmune en contra del bacilo. Actualmente se conocen distintas formas de presentación de la enfermedad, entre ellas, los polos determinados como el tubercúloide y lepromatoso, la forma indeterminada y la lepra borderline. El diagnostico temprano es la principal herramienta para lograr un tratamiento adecuado, prevenir las discapacidades y rehabilitar al paciente enfermo. El tratamiento fijado por la OMS desde 1984 consiste en una terapia multimedicamentosa basada en la clasificación paucibacilar o multibacilar, según el número de lesiones cutáneas y la cantidad de bacilos presentes en la biopsia. Se reporta el caso de un paciente femenino de 51 años, con diagnóstico de Lepra Borderline Lepromatosa que presentó reacción leprosa tipo 1 y alergia medicamentosa. Se discute las pruebas diagnosticas realizadas y la terapéutica empleada. (DUAZARY 2010, 71 - 78AbstractLeprosy is a chronic bacterial infection caused by Mycobacterium Leprae, an acid-fast bacillus forced to live in the intracellular space mainly in Schwann cells and macrophages, which affects the skin, nerves and eyes. The existence of the disease knows since antiquity, but still remains a serious public health problem worldwide especially in endemic areas. Its clinical spectrum is given by the response generated for the immune system against the bacillus. Today we know different forms of disease presentation, including the determined poles as the tuberculoid and

  8. ¿Es la resistencia de Mycobacterium leprae a los medicamentos un verdadero motivo de preocupación? Primera aproximación a la vigilancia molecular de pacientes colombianos multibacilares con tratamiento previo para lepra y sin él

    Directory of Open Access Journals (Sweden)

    Martha Inírida Guerrero

    2014-04-01

    Full Text Available Introducción. Colombia no dispone de información sobre farmacorresistencia primaria y secundaria de Mycobacterium leprae al esquema de terapia múltiple de la Organización Mundial de la Salud (OMS y las autoridades de salud pública del mundo han emitido varias recomendaciones, entre las cuales está organizar de inmediato la vigilancia a la resistencia empleando métodos moleculares simples. Objetivo. Determinar la prevalencia de la resistencia de M. leprae a rifampicina, ofloxacina y dapsona en pacientes del Centro Dermatológico Federico Lleras Acosta con tratamiento previo y sin él durante el período de 1985 a 2004. Materiales y métodos. Se realizó un estudio retrospectivo. Mediante muestreo electivo se incluyeron biopsias de pacientes multibacilares: 381 de pacientes nuevos y 560 de pacientes previamente tratados. Se obtuvieron con micrótomo seis cortes de cada biopsia de piel incluida en parafina, y se realizó la extracción de ADN de M. leprae. Se llevó a cabo la amplificación de tres blancos moleculares mediante PCR y se obtuvieron los patrones de resistencia a los medicamentos dapsona, rifampicina y ofloxacina por hibridación inversa. Se recolectaron datos epidemiológicos, clínicos y demográficos para llevar a cabo los análisis. Resultados. De las 941 muestras estudiadas, 4,14 % era resistente a uno o más fármacos, y se detectaron 5,77 y 3,04 % con genotipos resistentes en pacientes nuevos y previamente tratados, respectivamente. La resistencia total para cada fármaco fue de 0,43 % a dapsona, 3,19 % a rifampicina y 1,17 % a ofloxacina. Se encontró una diferencia estadísticamente significativa para rifampicina y para la población total al comparar los resultados de los pacientes no tratados con los de los pacientes tratados previamente. Dos tercios de las muestras resistentes lo fueron a rifampicina sola o combinada. Conclusiones. Los esquemas de terapia múltiple estándar siguen siendo efectivos para los casos de

  9. Papel del factor de crecimiento epidérmico recombinante humano “Heberprot-P” en un paciente con Lepra Multibacilar

    Directory of Open Access Journals (Sweden)

    Wilver Velázquez Chacón

    2014-08-01

    Full Text Available Se presenta el caso de un paciente masculino de 59 años de edad, tratado por vías convencionales a causa de lesiones ulcerativas de miembros inferiores, de más de siete años de evolución, sin diagnóstico y tratamiento. La evolución ulterior permitió reconocer la aparición subsecuente de lepra multibacilar. La basiloscopía fue positiva, indicando el diagnóstico de las lesiones ulcerativas, que se trataron con el uso de factor de crecimiento epidérmico recombinante humano en su formulación Heberprot-P, permitiendo la granulación de las lesiones en toda su extensión en un plazo corto, de 16 días, sin que se presentaran eventos adversos graves. Su resultado final fue satisfactorio

  10. La proteína asociada a SLAM (SAP regula la expresión de IFN-g en lepra The SLAM-associated protein (SAP regulates IFN-g expression in leprosy

    Directory of Open Access Journals (Sweden)

    María F. Quiroga

    2004-10-01

    Full Text Available La inmunidad protectora contra Mycobacterium leprae requiere IFN-g. Los pacientes con lepra tuberculoide producen localmente citoquinas Th1, mientras que los pacientes lepromatosos producen citoquinas Th2. La molécula linfocitaria activadora de señales (SLAM y la proteína asociada a SLAM (SAP participan en la diferenciación celular que conduce a producción de patrones específicos de citoquinas. A fin de investigar la vía SLAM/SAP en la infección por M. leprae, determinamos expresión de ARN mensajero (ARNm de SAP, IFN-g y SLAM en pacientes con lepra. Observamos que la expresión de SLAM correlacionó en forma directa con la expresión de IFN-g, mientras que la expresión de SAP correlacionó inversamente con la expresión de ambas proteínas. Así, nuestros resultados indican que SAP interferiría con las respuestas de citoquinas Th1 mientras que SLAM contribuiría con la respuesta Th1 en lepra, señalando a la vía SLAM/SAP como potencial blanco modulador de citoquinas en enfermedades con respuestas Th2 disfuncionales.Tuberculoid leprosy patients locally produce Th1 cytokines, while lepromatous patients produce Th2 cytokines. Signaling lymphocytic activation molecule (SLAM and the SLAM-associated protein (SAP participate in the differentiation process that leads to the production of specific patterns of cytokines by activated T cells. To investigate the SLAM/SAP pathway in M. leprae infection, we determined the expression of SAP, IFN-g and SLAM RNA messenger in leprosy patients. We found a direct correlation of SLAM expression with IFN-g expression, whereas the expression of SAP was inversely correlated with the expression of both SLAM and IFN-g. Therefore, our data indicate that SAP might interfere with Th1 cytokine responses while SLAM expression may contribute to Th1 responses in leprosy. This study further suggests that the SLAM/SAP pathway might be a focal point for therapeutic modulation of T cell cytokine responses in diseases

  11. Nota preliminar sobre la histopatología de las manifestaciones cutáneas de la "forma de lucio", de la lepra

    Directory of Open Access Journals (Sweden)

    Manuel Martínez Báez

    1942-03-01

    Full Text Available Por En el año de 1851, el ilustre médico mexicano don Rafael Lucio, en colaboración con el doctor don Ignacio Alvarado, publico su "Opúsculo sobre el Mal de San Lázaro o Elefanciasis de los griegos", en el cual, como se dice en  la introducción de tan notable obra, Lucio condensó el resultado de las observaciones hechas durante ocho años desde la dirección, a él encomendada, del "Hospital de los Lazarinos" en esta ciudad de México. EI opúsculo citado contiene la descripción de las formas de lepra estudiadas por el autor del mismo, con observaciones sagaces y curiosas sobre la evolución del padecimiento. EI autor pone especial interes en la descripción de una de las formas que suele revestir la lepra entre nosotros, "la que está caracterizada por la presencia de manchas rojas y dolorosas en la piel".  En 130 misma introducción, Don Rafael Lucio expresa que quiso que otro profesor en Medicina se asociase a él, por algunos meses, para observar a los pacientes, para "sacar 180 historias", de la mayor parte de los mismos y para que, comparando sus observaciones con las hechas, antes por el propio doctor Lucio, comprobase la verdad de lo asentado por este último. El opúsculo a que me refiero fue leído en esta Academia, en las sesiones de 31 de diciembre de 1851 y de enero de 1852.

  12. Paracoccidioidomicosis perianal asociada a lepra lepromatosa: Presentación de un caso clínico Perianal paracoccidioidomycosis associated with lepromatous leprosy: A clinical case report

    Directory of Open Access Journals (Sweden)

    M A Reyes

    2008-06-01

    Full Text Available Se presenta el caso de un paciente alcoholista con una ulceración perianal y manifestaciones cutáneas de enfermedad de Hansen. La biopsia de la lesión perianal y otros estudios arribaron al diagnóstico de una forma diseminada de paracoccidioidomicosis, así como también las biopsias cutáneas y los estudios baciloscópicos fueron diagnósticos de lepra lepromatosa. La respuesta a la terapéutica fue satisfactoria con desaparición de las lesiones cutáneas. La disminución de la respuesta inmunológica celular genera un terreno propicio para la infección de estos gérmenes y al compartir área endémica la asociación entre ambas patologías puede ocurrir.Lepra and Paracoccidioidomycosis are endemic diseases in Argentina. We report a case of a patient with an unusual perianal ulceration and cutaneous manifestations of Hansen's disease. The biopsy of perianal lesion and subsequent studies revealed a disseminated form of paracoccidioidomycosis, as well as skin biopsy and baciloscopic finding diagnostic of Lepromatous Leprosy. The main portal of entry of paracoccidioides is the lung. Hematogenous dissemination of the fungus may occur at this time, with the establishment of metastatic foci in any organ. Anal and perianal lesions are present only in 1.3 to 2.4% of the patients. The pathogenesis of anal lesions remains unclear, it may be secondary from a systemic or a local disease. The patient response to the therapeutic was notable, with disappearance of lesions up to the third month of started itraconazole orally 400 mg/day leading just atrophy scars in perianal areas. The treatment of Hansen's disease was made according to OMS guidelines for multibacillary disease.

  13. Construction of a ColD cda Promoter-Based SOS-Green Fluorescent Protein Whole-Cell Biosensor with Higher Sensitivity toward Genotoxic Compounds than Constructs Based on recA, umuDC, or sulA Promoters

    DEFF Research Database (Denmark)

    Norman, Anders; Hansen, Lars Hestbjerg; Sørensen, Søren Johannes

    2005-01-01

    -methyl-N'-nitro-N-nitrosoguanidine (MNNG) revealed that the promoter for the ColD plasmid-borne cda gene had responses 12, 5, and 3 times greater than the recA, sulA, and umuDC promoters, respectively, and also considerably higher sensitivity. Furthermore, we showed that when the SOS-GFP construct was introduced into an E. coli host...

  14. Contextos de abstinência e de recaída na recuperação da dependência química

    Directory of Open Access Journals (Sweden)

    Rigotto Simone Demore

    2002-01-01

    Full Text Available Foram entrevistados nove homens e três mulheres, residentes na região da cidade de Caxias do Sul - RS, todos diagnosticados como dependentes de substâncias, segundo critérios do DSM-IV, para que descrevessem suas experiências de abstinência e recaída nas tentativas de recuperação da dependência química. A análise qualitativa das entrevistas orientou-se pelos movimentos reflexivos de descrição, redução e interpretação fenomenológica. A experiência da abstinência foi atribuída aos seguintes constituintes e contextos experienciais: consciência do problema aditivo por parte do dependente, resgate de vínculos familiares, recomposição de auto-estima, afastamento de ambientes favorecedores da adição, e envolvimento em práticas religiosas. A ausência dos constituintes e contextos identificados na experiência de abstinência caracterizou a manutenção do consumo. Os elos experienciais interpretados como essenciais à experiência de abstinência foram as redes interpessoais de apoio - constituídas por profissionais, familiares e novos amigos - e o envolvimento como colaboradores na recuperação de outros dependentes químicos.

  15. Contextos de abstinência e de recaída na recuperação da dependência química

    Directory of Open Access Journals (Sweden)

    Simone Demore Rigotto

    Full Text Available Foram entrevistados nove homens e três mulheres, residentes na região da cidade de Caxias do Sul - RS, todos diagnosticados como dependentes de substâncias, segundo critérios do DSM-IV, para que descrevessem suas experiências de abstinência e recaída nas tentativas de recuperação da dependência química. A análise qualitativa das entrevistas orientou-se pelos movimentos reflexivos de descrição, redução e interpretação fenomenológica. A experiência da abstinência foi atribuída aos seguintes constituintes e contextos experienciais: consciência do problema aditivo por parte do dependente, resgate de vínculos familiares, recomposição de auto-estima, afastamento de ambientes favorecedores da adição, e envolvimento em práticas religiosas. A ausência dos constituintes e contextos identificados na experiência de abstinência caracterizou a manutenção do consumo. Os elos experienciais interpretados como essenciais à experiência de abstinência foram as redes interpessoais de apoio - constituídas por profissionais, familiares e novos amigos - e o envolvimento como colaboradores na recuperação de outros dependentes químicos.

  16. The Lon protease-like domain in the bacterial RecA paralog RadA is required for DNA binding and repair.

    Science.gov (United States)

    Inoue, Masao; Fukui, Kenji; Fujii, Yuki; Nakagawa, Noriko; Yano, Takato; Kuramitsu, Seiki; Masui, Ryoji

    2017-06-09

    Homologous recombination (HR) plays an essential role in the maintenance of genome integrity. RecA/Rad51 paralogs have been recognized as an important factor of HR. Among them, only one bacterial RecA/Rad51 paralog, RadA, is involved in HR as an accessory factor of RecA recombinase. RadA has a unique Lon protease-like domain (LonC) at its C terminus, in addition to a RecA-like ATPase domain. Unlike Lon protease, RadA's LonC domain does not show protease activity but is still essential for RadA-mediated DNA repair. Reconciling these two facts has been difficult because RadA's tertiary structure and molecular function are unknown. Here, we describe the hexameric ring structure of RadA's LonC domain, as determined by X-ray crystallography. The structure revealed the two positively charged regions unique to the LonC domain of RadA are located at the intersubunit cleft and the central hole of a hexameric ring. Surprisingly, a functional domain analysis demonstrated the LonC domain of RadA binds DNA, with site-directed mutagenesis showing that the two positively charged regions are critical for this DNA-binding activity. Interestingly, only the intersubunit cleft was required for the DNA-dependent stimulation of ATPase activity of RadA, and at least the central hole was essential for DNA repair function. Our data provide the structural and functional features of the LonC domain and their function in RadA-mediated DNA repair. © 2017 by The American Society for Biochemistry and Molecular Biology, Inc.

  17. La eliminación de la lepra de las Américas: situación actual y perspectivas

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    Clovis Lombardi

    1998-09-01

    Full Text Available La lepra, enfermedad que antes evocaba una imagen sombría e inspiraba terror, ahora se puede curar gracias al esquema politerapéutico a base de rifampicina, clofazimina y dapsona que se ha venido usando desde 1981. En 1991 la Asamblea Mundial de la Salud, alentada por la eficacia de este régimen, fijó la meta de eliminar la enfermedad como problema de salud pública mundial y nacional para el año 2000. Esta meta, que equivale a reducir la prevalencia a menos de un caso por 10 000 habitantes, no debe confundirse con la de erradicar la enfermedad, que implica interrumpir por completo su transmisión. La eliminación de la lepra es una meta asequible que dependerá del uso enérgico y a gran escala del régimen poliquimioterapéutico. El presente trabajo describe y examina las iniciativas que se han puesto en marcha en América Latina para lograr la meta y los resultados observados hasta el momento. También se exploran los factores que inciden en la factibilidad de erradicar la enfermedad.Leprosy, a disease that used to be shrouded in darkness and fear, can now be cured thanks to a multidrug treatment schedule with rifampicin, clofazimine, and dapsone which has been in use since 1981. In 1991 the World Health Assembly, enouraged by the efficacy of this treatment regimen, established the goal of eliminating the disease as a public health problem globally and nationally by the year 2000. This goal, which calls for reducing disease prevalence to less than one case per 10 000 inhabitants, should not be confused with the goal of eradicating the disease, which implies a complete interruption of its transmission. Eliminating leprosy is an attainable goal which will depend on the forceful and massive use of the multidrug treatment regimen. This paper describes and discusses the various initiatives that have been launched in Latin America for the purpose of achieving this goal and the results obtained so far. It also explores the factors that impact

  18. Estudo de caso: prevenção de recaída para criança e adolescente enuréticos com remissão espontânea

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    Rodrigo Fernando Pereira

    2006-08-01

    Full Text Available A literatura aponta o tratamento comportamental com alarme como o mais eficaz para os casos de enurese noturna. Quando um procedimento comportamental adicional, como a superaprendizagem, é adicionado a este tratamento, as taxas de recaída são reduzidas. Neste procedimento há um gradual aumento de ingestão de água imediatamente antes de dormir. Possivelmente o maior controle obtido deve-se a essa combinação atuar tanto nos níveis plasmáticos de vasopressina como na capacidade funcional da bexiga. Os dois casos apresentados são de enuréticos que haviam adquirido o controle recentemente, mas suas famílias demandavam um atendimento para diminuir as chances de recaída. Utilizou-se, então, o alarme conjugado à superaprendizagem desde o início do atendimento, com a finalidade de se estabelecer um controle completo. No primeiro caso, de uma adolescente de dezoito anos, não houve molhadas durante o tratamento. No segundo, de um garoto de oito anos, as molhadas voltaram com a ingestão de líquido e tornaram a cessar, possivelmente por ter sido obtido um controle maior por parte da criança. Acredita-se que o procedimento descrito possa ser útil nos casos de crianças que deixaram recentemente de ser enuréticas ou não alcançam o critério para diagnóstico. Palavras-chave: enurese noturna; prevenção de recaída; superaprendizagem.

  19. Metodologia diagnóstica e tratamento da recaída após transplante de medula óssea em paciente com leucemia promielocítica aguda

    Directory of Open Access Journals (Sweden)

    Pallotta Ronald

    2000-01-01

    Full Text Available O transplante de medula óssea, que é um consagrado tratamento de várias doenças oncológicas, hematológicas e imunológicas, tem a recaída como um das principais causas de falha. As variáveis de seleção, condicionamento e de seguimento devem ser analisadas, pois influenciam de maneira fundamental tal falha. O diagnóstico precoce da recaída tem como pilar de sustentação a capacidade de detecção a doença residual mínima. A metodologia empregada, com a citometria de fluxo, citogénetica convencional, fluorescent in situ hybridization e com o polimerase chain reaction, permite uma detecção cada vez mais precoce deste clone residual que é responsável pela volta da doença de base. L.M.S.O, de 54 anos, teve o diagnóstico de leucemia promielocítica em 1997. Foi submetida a um transplante de medula óssea autogênico em 01/09/98 como terapêutica de consolidação. Dois anos após o transplante de medula óssea apresentou recaída molecular diagnosticada por RT PCR realizado sequencialmente. O uso de ácido all-trans-retinóico foi capaz de induzir a nova remissão molecular. O all-trans-retinóico, que tem um mecanismo de ação não bem estabelecido na indução da remissão completa na LMA M3, pode ser utilizado em recaídas. No caso apresentado o diagnóstico precoce a partir de um esquema de avaliação pós-transplante bem programado permitiu um bom resultado. Conclui-se que as variáveis de seleção, condicionamento e seguimento devem ser muito bem estudadas para planejar um acompanhamento adequado e prolongado após o transplante de medula óssea, permitindo cada vez mais melhores resultados com o procedimento.

  20. Educación en la promoción de estilos de vida saludables para la prevención de recaídas en pacientes con tuberculosis pulmonar bk (+), en el hospital Carlos Monge Medrano Juliaca 2010.

    OpenAIRE

    Maldonado Zanabria, Dionicia

    2011-01-01

    El presente estudio se realizó con el objetivo de evaluar la efectividad de la Educación en la Promoción de estilos de vida saludable para la prevención de recaídas en pacientes con Tuberculosis Pulmonar BK (+) en el Hospital Carlos Monge Medran o Juliaca 2010. El estudio es de carácter Cuasi-Experimental, con diseño de pre y post test; la muestra estuvo constituida por 18 pacientes. Para recolección de datos se utilizó como instrumentos un cuestionario. La prueba de hipótesis se realizó con ...

  1. Antígeno soluble Mycobacterium leprae y PPD en escolares de la Isla de Pascua y Santiago de Chile, 1987

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    Hugo Hurtado G.

    1989-12-01

    Full Text Available Se midió la reacción cruzada de los antígenos proteina pura derivada de tuberculina (PPD y antígeno soluble de Mycobacterium leprae (ASML mediante dos muestras de niños entre 6-14 años: una en un área endémica (Isla de Pascua y la otra en una no endémica (Santiago de Chile continental. Cada grupo de niños se inoculó simultáneamente con PPD y ASML en los antebrazos derecho e izquierdo respectivamente. Las lecturas fueron hechas 48 horas después. Los análisis estadísticos mostraron una evidente reacción cruzada de los dos antígenos. Se presentó también una significativa mayor reacción en el área endémica (Pascua que en Santiago tanto para PPD como para ASML.

  2. Research Article Special Issue

    African Journals Online (AJOL)

    2016-05-15

    May 15, 2016 ... (NMR), immunological properties survey, enzymology and medical applications such as pharmaceutical proteins as well as ... ELIZA1 (Enzyme –linked immune sorbent assay) and quantitative study (Murby M,. 1991). These tags are areas in ..... Mycobacterium tuberculosis (mtu RecA). This Intein is applied ...

  3. Fatores de risco e proteção à recaída na percepção de usuários de substâncias psicoativas

    Directory of Open Access Journals (Sweden)

    Meire Luci da Silva

    2014-01-01

    Full Text Available El objetivo de este estudio fue identificar, en la percepción de usuarios de sustancias psicoactivas, factores de riesgo y protección a la recaída. Participaron 50 usuarios en tratamiento en una Comunidad Terapéutica, en São Paulo, Brasil, en 2013. Investigación cuantitativa con cuestionarios auto administrado con preguntas cerradas y análisis a través de estadística descriptiva. Se identificaron como factores de riesgo: falta de apoyo familiar, conflictos familiares, sentimientos negativos, contexto social, abandono de grupos de apoyo, insatisfacción con tratamiento y dificultades financieras. Factores de protección destacados: religiosidad y grupos de apoyo, siendo el apoyo profesional una de las últimas redes de apoyo. Se constató ambigüedad de familia y amigos mientras factor de riesgo y protección. Se espera que factores de riesgos y protección identificados contribuyan para políticas de prevención a recaídas, permitiendo mejoramiento de tratamientos centrados en reconocimiento de factores de protección, desarrollo de habilidades y estrategias de afrontamiento.

  4. Expression of Mycobacterium tuberculosis Ku and Ligase D in Escherichia coli results in RecA and RecB-independent DNA end-joining at regions of microhomology.

    Science.gov (United States)

    Malyarchuk, Svitlana; Wright, Douglas; Castore, Reneau; Klepper, Emily; Weiss, Bernard; Doherty, Aidan J; Harrison, Lynn

    2007-10-01

    Unlike Escherichia coli, Mycobacterium tuberculosis (Mt) expresses a Ku-like protein and an ATP-dependent DNA ligase that can perform non-homologous end-joining (NHEJ). We have expressed the Mt-Ku and Mt-Ligase D in E. coli using an arabinose-inducible promoter and expression vectors that integrate into specific sites in the E. coli chromosome. E. coli strains have been generated that express the Mt-Ku and Mt-Ligase D on a genetic background that is wild-type for repair, or deficient in either the RecA or RecB protein. Transformation of these strains with linearized plasmid DNA containing a 2bp overhang has demonstrated that expression of both the Mt-Ku and Mt-Ligase D is required for DNA end-joining and that loss of RecA does not prevent this double-strand break repair. Analysis of the re-joined plasmid has shown that repair is predominantly inaccurate and results in the deletion of sequences. Loss of RecB did not prevent the formation of large deletions, but did increase the amount of end-joining. Sequencing the junctions has revealed that the majority of the ligations occurred at regions of microhomology (1-4bps), eliminating one copy of the homologous sequence at the junction. The Mt-Ku and Mt-Ligase D can therefore function in E. coli to re-circularize linear plasmid.

  5. Implication of the E. coli K12 uvrA and recA genes in the repair of 8-methoxypsoralen-induced mono adducts and crosslinks on plasmid DNA

    International Nuclear Information System (INIS)

    Paramio, J.M.; Bauluz, C.; Vidania, R. de

    1986-01-01

    Genotoxicity of psoralen damages on plasmid DNA has been studied. pBR322 DNA was randomly modified with several concentrations of 8-methoxypsoralen plus 365 nm-UV light. After transformation into E. coli strains (wild-type, uvrA and recA) plasmid survival and mutagenesis were analyzed. To study the influence of the SOS response on plasmid recovery, preirradiation of the cells was performed. In absence of cell preirradiation, crosslinks were not repaired in any strain. Mono adducts were also lethal but in part removed by the excision-repair pathway. Preirradiation of the cells significantly. increased plasmid recovery in recA+ celia. In uvrA- only the mutagenic pathway seemed to be involved in the repair of the damaged DNA. Wild type strain showed the highest increase in plasmid survival, involving the repair of mono adducts and some fraction of crosslinks mainly through an error-free repair pathway. This suggests an enhancement of the excision repair promoted by the induction of SOS functions. (Author) 32 refs

  6. Cloning of an E. coli RecA and yeast RAD51 homolog, radA, an allele of the uvsC in Aspergillus nidulans and its mutator effects.

    Science.gov (United States)

    Seong, K Y; Chae, S K; Kang, H S

    1997-04-30

    An E. coli RecA and yeast RAD51 homolog from Aspergillus nidulans, radA, has been cloned by screening genomic and cDNA libraries with a PCR-amplified probe. This probe was generated using primers carrying the conserved sequences of eukaryotic RecA homologs. The deduced amino acid sequence revealed two conserved Walker-A and -B type nucleotide-binding domains and exhibited 88%, 60%, and 53% identity with Mei-3 of Neurospora crassa, rhp51+ of Schizosaccharomyces pombe, and Rad51 of Saccharomyces cerevisiae, respectively. radA null mutants constructed by replacing the whole coding region with a selection marker showed high methyl methanesulfonate (MMS) sensitivity. Heterozygous diploids of radA disruptant with the uvsC114 mutant failed to complement with respect to MMS-sensitivity, indicating that radA is an allele of uvsC. In selecting spontaneous forward selenate resistant mutations, mutator effects were observed in radA null mutants similarly to those shown in uvsC114 mutant strains.

  7. The crystal structure of M. leprae ML2640c defines a large family of putative S-adenosylmethionine-dependent methyltransferases in mycobacteria

    Science.gov (United States)

    Graña, Martin; Haouz, Ahmed; Buschiazzo, Alejandro; Miras, Isabelle; Wehenkel, Annemarie; Bondet, Vincent; Shepard, William; Schaeffer, Francis; Cole, Stewart T.; Alzari, Pedro M.

    2007-01-01

    Mycobacterium leprae protein ML2640c belongs to a large family of conserved hypothetical proteins predominantly found in mycobacteria, some of them predicted as putative S-adenosylmethionine (AdoMet)-dependent methyltransferases (MTase). As part of a Structural Genomics initiative on conserved hypothetical proteins in pathogenic mycobacteria, we have determined the structure of ML2640c in two distinct crystal forms. As expected, ML2640c has a typical MTase core domain and binds the methyl donor substrate AdoMet in a manner consistent with other known members of this structural family. The putative acceptor substrate-binding site of ML2640c is a large internal cavity, mostly lined by aromatic and aliphatic side-chain residues, suggesting that a lipid-like molecule might be targeted for catalysis. A flap segment (residues 222–256), which isolates the binding site from the bulk solvent and is highly mobile in the crystal structures, could serve as a gateway to allow substrate entry and product release. The multiple sequence alignment of ML2640c-like proteins revealed that the central α/β core and the AdoMet-binding site are very well conserved within the family. However, the amino acid positions defining the binding site for the acceptor substrate display a higher variability, suggestive of distinct acceptor substrate specificities. The ML2640c crystal structures offer the first structural glimpses at this important family of mycobacterial proteins and lend strong support to their functional assignment as AdoMet-dependent methyltransferases. PMID:17660248

  8. Apreciaciones críticas a los programas de control y eliminación de la lepra en el Perú y sus consecuencias para el Perú y América

    Directory of Open Access Journals (Sweden)

    Zuño Burstein

    Full Text Available Se presenta un análisis crítico de las medidas de control sanitario que, históricamente, se llevaron a cabo en el Perú hasta llegar a la situación actual, que sitúa al Perú en la fase oficialmente considerada de “eliminación” de la lepra como problema de salud pública. Se discute, además, su validación, señalándose que el descuido de la vigilancia sanitaria, la desarticulación de los organismos de control especializado, el olvido de parte del personal médico sanitario de su diagnóstico precoz, la presencia de casos lepromatosos bacilíferos no diagnosticados en la capital de la república y la indudable prevalencia escondida, permiten concluir que existe el peligro para el país y la región que se reproduzca una recrudescencia de la lepra, si no se toman las medidas sanitarias pertinentes y oportunas

  9. As dimensões da sustentabilidade dos Sistemas Agroflorestais – SAFs: um estudo no Projeto de Reflorestamento Consorciado e Adensado – RECA, Ponta do Abunã – RO

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    Ana Íris Tomás Vasconcelos

    2016-04-01

    Full Text Available The study aimed to determine the contribution of Joint Venturer for Economic Reforestation (Reflorestamento Econômico Consorciado e Adensado, RECA, which is an agroforestry system, organized in communities which produce peach palm, Brazil nut, cupuaçu and other forest products from the perspective of four sustainability dimensions: institutional, economic, environmental and social. The methodology chosen, was a content analysis through the evaluation of the project field notes, which were analyzed using the software AtlasTi in the light of the categories that were established and presented, considering those four dimensions. After analyzing the results, they showed that there was: improvement in the associates incomes, maturing of the community for collective work, sustainability-oriented practices, performance differences among the group members as well as product and byproducts development. After observing positive impacts on the aspect of sustainability, the carry out of additional studies to assess the institutional dimension in the project was suggested.

  10. Resistance to nitrofurantoin and UV-irradiation in recA; uvrA; and uvrA, lexA, Escherichia coli mutants conferred by an R-plasmid from an Escherichia coli clinical isolate

    International Nuclear Information System (INIS)

    Obaseiki-Ebor, E.E.

    1984-01-01

    There have been some reports of R-plasmids conferring nitrofuran resistance by decreasing the reduction of nitrofurantoin. The mechanism by which these R-plasmids mediate nitrofurantoin resistance is still not properly understood. Since DNA repair mutants are very sensitive to nitrofurantoin, it was therefore of interest to see whether R-plasmids conferring nitrofurantoin resistance affected the nitrofurantoin sensitivity of recA; uvrA and uvrA, lexA strains of E. coli K-12. Protection against UV-irradiation was also estimated. The experiments showed that the nitrofurantoin resistance conferred by R-plasmid pBN105 was not due to defective nitrofurantoin reduction or altered permeability of the cell. Because it is known that repair-deficient bacteria have increased susceptibility to nitrofurantoin, it may be suggested that the mechanisms of UV and nitrofurantoin protection conferred by pBN105 to the DNA repair mutant strains are related. (Auth.)

  11. Mycobacterium tuberculosis nucleoid-associated DNA-binding protein H-NS binds with high-affinity to the Holliday junction and inhibits strand exchange promoted by RecA protein.

    Science.gov (United States)

    Sharadamma, N; Harshavardhana, Y; Singh, Pawan; Muniyappa, K

    2010-06-01

    A number of studies have shown that the structure and composition of bacterial nucleoid influences many a processes related to DNA metabolism. The nucleoid-associated proteins modulate not only the DNA conformation but also regulate the DNA metabolic processes such as replication, recombination, repair and transcription. Understanding of how these processes occur in the context of Mycobacterium tuberculosis nucleoid is of considerable medical importance because the nucleoid structure may be constantly remodeled in response to environmental signals and/or growth conditions. Many studies have concluded that Escherichia coli H-NS binds to DNA in a sequence-independent manner, with a preference for A-/T-rich tracts in curved DNA; however, recent studies have identified the existence of medium- and low-affinity binding sites in the vicinity of the curved DNA. Here, we show that the M. tuberculosis H-NS protein binds in a more structure-specific manner to DNA replication and repair intermediates, but displays lower affinity for double-stranded DNA with relatively higher GC content. Notably, M. tuberculosis H-NS was able to bind Holliday junction (HJ), the central recombination intermediate, with substantially higher affinity and inhibited the three-strand exchange promoted by its cognate RecA. Likewise, E. coli H-NS was able to bind the HJ and suppress DNA strand exchange promoted by E. coli RecA, although much less efficiently compared to M. tuberculosis H-NS. Our results provide new insights into a previously unrecognized function of H-NS protein, with implications for blocking the genome integration of horizontally transferred genes by homologous and/or homeologous recombination.

  12. Evolutionary relationships among salivarius streptococci as inferred from multilocus phylogenies based on 16S rRNA-encoding, recA, secA, and secY gene sequences

    Directory of Open Access Journals (Sweden)

    Boissinot Maurice

    2009-10-01

    Full Text Available Abstract Background Streptococci are divided into six phylogenetic groups, i.e, anginosus, bovis, mitis, mutans, pyogenic, and salivarius, with the salivarius group consisting of only three distinct species. Two of these species, Streptococcus salivarius and Streptococcus vestibularis, are members of the normal human oral microflora whereas the third, Streptococcus thermophilus, is found in bovine milk. Given that S. salivarius and S. vestibularis share several physiological characteristics, in addition to inhabiting the same ecosystem, one would assume that they would be more closely related to each other than to S. thermophilus. However, the few phylogenetic trees published so far suggest that S. vestibularis is more closely related to S. thermophilus. To determine whether this phylogenetic relationship is genuine, we performed phylogenetic inferences derived from secA and secY, the general secretion housekeeping genes, recA, a gene from a separate genetic locus that encodes a major component of the homologous recombinational apparatus, and 16S rRNA-encoding gene sequences using other streptococcal species as outgroups. Results The maximum likelihood (ML and maximum parsimony (MP phylogenetic inferences derived from the secA and recA gene sequences provided strong support for the S. vestibularis/S. thermophilus sister-relationship, whereas 16S rRNA-encoding and secY-based analyses could not discriminate between alternate topologies. Phylogenetic analyses derived from the concatenation of these sequences unambiguously supported the close affiliation of S. vestibularis and S. thermophilus. Conclusion Our results corroborated the sister-relationship between S. vestibularis and S. thermophilus and the concomitant early divergence of S. salivarius at the base of the salivarius lineage.

  13. Evolutionary relationships among salivarius streptococci as inferred from multilocus phylogenies based on 16S rRNA-encoding, recA, secA, and secY gene sequences.

    Science.gov (United States)

    Pombert, Jean-François; Sistek, Viridiana; Boissinot, Maurice; Frenette, Michel

    2009-10-30

    Streptococci are divided into six phylogenetic groups, i.e, anginosus, bovis, mitis, mutans, pyogenic, and salivarius, with the salivarius group consisting of only three distinct species. Two of these species, Streptococcus salivarius and Streptococcus vestibularis, are members of the normal human oral microflora whereas the third, Streptococcus thermophilus, is found in bovine milk. Given that S. salivarius and S. vestibularis share several physiological characteristics, in addition to inhabiting the same ecosystem, one would assume that they would be more closely related to each other than to S. thermophilus. However, the few phylogenetic trees published so far suggest that S. vestibularis is more closely related to S. thermophilus. To determine whether this phylogenetic relationship is genuine, we performed phylogenetic inferences derived from secA and secY, the general secretion housekeeping genes, recA, a gene from a separate genetic locus that encodes a major component of the homologous recombinational apparatus, and 16S rRNA-encoding gene sequences using other streptococcal species as outgroups. The maximum likelihood (ML) and maximum parsimony (MP) phylogenetic inferences derived from the secA and recA gene sequences provided strong support for the S. vestibularis/S. thermophilus sister-relationship, whereas 16S rRNA-encoding and secY-based analyses could not discriminate between alternate topologies. Phylogenetic analyses derived from the concatenation of these sequences unambiguously supported the close affiliation of S. vestibularis and S. thermophilus. Our results corroborated the sister-relationship between S. vestibularis and S. thermophilus and the concomitant early divergence of S. salivarius at the base of the salivarius lineage.

  14. Prevención de recaídas en la esquizofrenia: propuesta de un programa de intervención durante la fase prodrómica (Relapse prevention in schizophrenia: a proposal of intervention during the prodromal phase

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    Juan Francisco Godoy García

    2016-04-01

    Full Text Available La esquizofrenia es un trastorno psicológico crónico y grave que afecta al 1% de la población mundial adulta. El curso del pródromo comienza en la mayoría de los casos con la experimentación de síntomas clínicos negativos, seguidos de síntomas positivos, que se van agravando en frecuencia, duración e intensidad conforme se aproxima el episodio de psicosis. Se propone una intervención multicomponente destinada a pacientes con esquizofrenia y a sus familiares, con 24 sesiones dirigidas a la psicoeducación y adherencia al tratamiento, detección e intervención sobre signos tempranos y fortalecimiento de la autoeficacia. Dicha intervención pretende ayudarles a detectar y modificar los principales marcadores prodrómicos, entrenando a los pacientes y a sus familiares en habilidades útiles para prevenir o afrontar la recaída, con el fin de evitar o retrasar la nueva crisis psicótica. Consecuentemente, cabe esperar que los pacientes mejoren la comprensión de su enfermedad y su adherencia a la medicación, reconozcan y actúen sobre los primeros signos de la posible recaída, incrementen su autoeficacia, se reduzcan las tasas de recaída y, en definitiva, mejore tanto su estado clínico como su calidad de vida. En cuanto a los familiares, se persigue, por un lado, ayudar al paciente en todo lo anterior y, por otro, aliviar su carga como cuidadores, mejorando su calidad de vida. Abstract: Schizophrenia is a chronic, severe mental disorder affecting 1% of the adult population worldwide (WHO, 2014. The course of the prodromal stage generally begins with experiencing negative clinical symptoms, followed by positive symptoms, which usually increase in frequency, duration and intensity as the episode of psychosis approaches. A 24-session multi-component intervention -psychoeducation and adherence to treatment, early detection and intervention on initial signs and self-efficacy strengthening- is proposed herein, which is aimed at patients

  15. A monument to Lazarus: the leprosy hospital of Rio de Janeiro Um monumento a Lázaro: o hospital de lepra do Rio de Janeiro

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    Thomas Hunter Smith III

    2003-01-01

    Full Text Available Soon after the Portuguese made landfall in 1500, Europeans and, later, African slaves introduced leprosy, and Saint Lazarus, the patron saint of its victims, into Brazil. Social and political pressure mounted by the middle of the eighteenth century in the city of Rio de Janeiro to remove those unfortunates from the city's streets even before the move of Brazil's capital in 1763. Frei Antônio, the bishop of Rio, founded the venerable hospital that year in the neighborhood of São Cristóvão. He requested that the Irmandade do Santíssimo Sacramento da Candelária provide oversight and administration. The brotherhood continues to honor its covenant of 239 years ago. The history of this hospital provides insight into the complex relationships that existed between the citizenry and church and state. Rio's leprosy hospital, now the Hospital Frei Antônio, had an important role in the evolution of the health care professions, progress in medical science, and the genesis of the hygienic movement in Brazil. This study also contributes to the history of a disease that persists in 2002 Brazil as a public health issue.Logo após a chegada dos portugueses, em 1500, os europeus e, mais tarde, os escravos africanos introduziram a lepra no Brasil e, com ela, São Lázaro, o patrono de suas vítimas. No Rio de Janeiro, cresceram as pressões para que as vítimas fossem removidas das ruas da cidade, antes mesmo da mudança da capital do Brasil, em 1763. Neste ano, Frei Antônio, o bispo do Rio de Janeiro, fundou o venerável hospital no bairro de São Cristóvão. Solicitou que a Irmandade do Santíssimo Sacramento da Candelária cuidasse da assistência e administração do hospital. A irmandade continua a honrar o acordo feito há 239 anos. A história desse hospital é uma fonte de conhecimento sobre as relações complexas existentes entre os cidadãos, a Igreja e o Estado. O hospital atualmente chamado Frei Antônio teve importante papel na evolução de

  16. Fatores de risco para transmissão da Hanseníase Factores de riesgo para la transmisión de la Lepra Risk factors for Leprosy transmission

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    Andréia Soprani dos Santos

    2008-11-01

    Full Text Available Estudo caso controle que objetivou identificar fatores individuais de risco relacionados à transmissão da doença. O grupo caso, composto por 90 pacientes de hanseníase notificados no SINAN entre 2003 e 2006; o grupo controle, constituído por 270 indivíduos sadios, pareados por sexo e faixa etária. Houve associação significativa entre a ocorrência da doença e a presença atual (OR= 2,9 e antiga (OR=5,0 de hanseníase entre parentes co-sangüíneos. Sabendo-se que o exame único dos contatos, no ato do diagnóstico, detecta uma parcela mínima dos casos, propõe-se a realização de exames periódicos dos contatos de hansenianos a fim de detectar os novos casos.Estudio caso controle que objetivó identificar factores individuales del riesgo relacionados a transmisión de la enfermedad. El grupo caso, compuesto por 90 pacientes con lepra notificados en el SINAN entre 2003 y 2006; y el grupo controle, constituido por 270 individuos saludables, pareados por sexo y edad. Hubo asociación significativa entre la ocurrencia de la enfermedad y la presencia actual (OR= 2,9 y antigua (OR=5,0 de la lepra entre parientes co-sanguíneos. Sabiéndose que el examen único de los contactos en el acto del diagnóstico detecta una parcela mínima de los casos, se propone a la realización del examen periódico de los contactos de leprosos a fin de detectar los nuevos casos.This case-control study aimed to identify individual risk factors regarding the transmission of leprosy. The group case represented by 90 cases of leprosy sick people pontificated at SINAN during 2003 and 2006 and a group control constituted by 270 healthy people, paired by gender and age. There was significant statistical between occurrence of the disease and its current presence (OR: 2,9 and old cases (OR= 5,0 of leprosy among co-sanguine relatives. Knowing that the only exam of the contact in the act of the diagnostic detects a minimum part of the new cases, it is proposed to

  17. Epidemiología de la lepra a través del estudio de la frecuentación de el hospital especializado de Trillo durante el período 1943-1995

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    Urbina Torija Juan Román

    1997-01-01

    Full Text Available FUNDAMENTO: El estudio pretende describir el perfil sociodemográfico y clínico del enfermo de lepra hospitalizado y comprobar si su tipología ha cambiado en la historia del centro. MÉTODOS: Se trata de un estudio descriptivo y retrospectivo mediante la revisión de una muestra de las historias clínicas abiertas en el centro desde su origen en 1943 hasta 1995. Se eligieron mediante muestreo aleatorio sistemático 366 historias, de las que se cumplimentó un cuestionario que recogía variables sociodemográficas y clínicas. RESULTADOS: La mayoría de los enfermos fueron varones (71.9%, IC: 67.3-76.4, de edad media de 39.6 años (IC: 37.9-41.4 solteros (46.2%, IC: 41-51.3 con bajo nivel cultural (analfabetismo: 54.1%, IC: 40-69 y una ocupación relacionada con la agricultura (35.5%, IC:30.6-40.4 en regiones del sur de España (enfermos procedentes de Andalucía 52.8%, IC: 45.8-54. La enfermedad presentó antecedentes familiares en un 31.1% (IC:26.4-35.9 de los afectados y formas clínicas graves multibacilares (lepra lepromatosa 66.1%, IC:61.2-71 que afectaba a miembros inferiores en un 72.1%, cabeza en el 63.1% y miembros superiores 64.4%. La mortalidad global fue de 31.1% en los ingresados (IC: 26.4-35.9. A lo largo del período de estudio el enfermo ingresado ha envejecido y suavizado su clínica y mortalidad. La estancia hospitalaria ha sido de 7.1 años (IC: 6.1-8.1 en general, aunque al final del estudio se aproxima a tiempos de ingreso menores, acordes con la duración del tratamiento de la enfermedad (2.2 años. CONCLUSIONES: Un varó, joven, con economía precarias y residente en el sur de nuestro país, parece ser el perfil del enfermo, que con formas anatomoclínicas severas inicialmente y una evolución posterior favorable, puede representar esta enfermedad en un país con una endemia autóctona y una epidemiología característica, que está en fase de pre-erradicaciòn.

  18. Ensaio clínico randomizado e metanálise sobre a eficácia da buspirona e da carbamazepina no tratamento de prevenção de recaìdas em pacientes com dependência alcoólica

    OpenAIRE

    Santos, Sandro Iêgo da Silva

    2013-01-01

    Introdução. Poucos estudos investigaram os potenciais benefícios da carbamazepina e da buspirona no tratamento de prevenção de recaídas do alcoolismo, e o efeito desses fármacos sobre o consumo de álcool permanece inconcluso. Ademais, os tratamentos farmacológicos preconizados atualmente não são bem sucedidos em todos os casos, restando grupos de pacientes que não se beneficiam dos tratamentos disponíveis, para os quais essas drogas poderiam ser uma possível alternativa tera...

  19. Assistência de enfermagem ao portador de Hanseníase: abordagem transcultural Asistencia de enfermería al portador de Lepra: abordaje transcultural Nursing assistance to a Leprosy-infected patient: transcultural approach

    Directory of Open Access Journals (Sweden)

    Fernando José Guedes da Silva Júnior

    2008-11-01

    Full Text Available Trata-se de um estudo de caso, desenvolvido em um Centro de Saúde em Teresina - PI, que abordou a problemática da assistência de enfermagem prestada a uma paciente com Hanseníase Multibacilar, enfocando o cuidado de Enfermagem Transcultural, Diagnósticos e Intervenções de Enfermagem segundo a Taxonomia II da NANDA. Realizou-se a entrevista semi-estruturada e observação participante, que possibilitaram a coleta dos dados, os quais foram tratados baseados nos padrões normativos, valores e prática diárias, modos de cuidado popular e cuidados requeridos no sistema profissional. Neste estudo, constatou-se que o planejamento da assistência de enfermagem visou, principalmente, contribuir para a adesão ao tratamento da doença, diminuindo os riscos potenciais e utilizando a preservação, negociação e re-padronização do sistema profissional. Observou-se também a adesão ao tratamento e a realização de autocuidado voltado para a hanseníase de forma culturalmente satisfatória.Es un estudio de caso, desarrollado en un centro de salud en Teresina-PI, que abordó la problemática de la asistencia de enfermería dada a una paciente con lepra multibacilar, enfocando el cuidado de Enfermería Transcultural, Diagnósticos, e Intervenciones de Enfermería segundo la Taxonomía II de NANDA. Se realizó una entrevista semi-estructurada y observación participante, que posibilitaron la colecta de los datos, los cuales fueron tratados basados em los padrones normativos, valores y prácticas diárias, modos de cuidado popular y cuidados requeridos en el sistema profesional. En este estudio, se constató que el planeamiento de la asistencia de enfermería objetivó, principalmente, contribuir para la adhesión al tratamiento de la enfermidad, disminuyendo los riscos potenciales y utilizando la preservación, negociación y repradonización del sistema profesional. Se observó también la adhesión al tratamiento y la realización del auto cuidado

  20. Muscle involvement in leprosy: study of the anterior tibial muscle in 40 patients Alterações musculares na lepra: estudo do músculo tibial anterior em 40 pacientes

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    LINEU CESAR WERNECK

    1999-09-01

    Full Text Available The involvement of skeletal striated muscle in leprosy is considered secondary due to peripheral neuropathy, but some studies point it to a primary muscle lesion. In order to investigate the muscle involvement in leprosy, we studied 40 patients (lepromatous 23, tuberculoid 13, borderline 2 and indeterminate 2. The motor nerve conduction of the peroneal nerves had a reduction of the velocity, decreased compound muscle action potential and sometimes absence of potentials. The electromyographic study of the anterior tibial muscle showed signs of recent and chronic denervation in 77.5% of the cases and no myopathic potentials. The anterior tibial muscle biopsy revealed denervation in 45% of the cases, interstitial inflammatory myopathy in 30% and mixed (myopathic and neuropathic pattern in 12.5%. Acid fast bacillus was detected in 25% of the cases, always in the interstitial tissue. Inflammatory reaction was present in the interstitial space and in patients with the lepromatous type. The histological findings clearly defined the presence of the so-called "Leprous Interstitial Myositis" on the top of denervation signs.O envolvimento do músculo estriado na lepra é considerado secundário à lesão dos nervos periféricos, mas alguns estudos relataram acometimento muscular primário. A fim de verificar esta controvérsia estudamos 40 pacientes com lepra, sendo 23 da forma lepromatosa, 13 da tuberculoide, 2 borderline e 2 indeterminada. Realizamos a neurocondução do nervo peroneiro, junto com eletromiografia e biópsia do músculo tibial anterior. Encontramos redução de velocidade de condução, da amplitude e algumas vezes ausência de potenciais no nervo peroneiro. A eletromiografia do tibial anterior mostrou sinais de desinervação recente e crônica em 77,5% dos casos e não foi encontrada evidência de padrão "miopático". A biópsia do músculo tibial anterior revelou desinervação em 45% dos casos, miopatia inflamatória intersticial em

  1. Avaliação e controle de contatos faltosos de doentes com hanseníase Evaluación y control de contactos faltantes de enfermos con lepra Evaluation and control of missing contacts of leprosy patients

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    Carmen Silvia de Campos Almeida Vieira

    2008-11-01

    Full Text Available Estudo descritivo, com variáveis qualitativas, realizado no Ambulatório Regional de Especialidades de Taubaté, com objetivo de resgatar e avaliar contatos faltosos de doentes de hanseníase. Analisou-se 36 prontuários de casos novos, no período de janeiro de 2003 a julho de 2004. Identificou-se 92 contatos, sendo 64,1% faltosos, tendo 25% entre 20 a 29 anos e 58,5% do sexo feminino. Destes, 61% foram submetidos à consulta de enfermagem no domicilio. Relataram não adesão ao controle por esquecimento (66,6%, e falta de tempo (11,1%; 4 contatos tinham sintomas de hanseníase, e 1 foi confirmado (forma transmissível - Dimorfa. Sem atingir 100% da cobertura de busca ativa, sugerimos, para assegurar a meta de eliminação da doença, parcerias e descentralização das ações de controle.Estudio descriptivo, con cualitativas variable, realizado en el Ambulatorio de Especialidades Regional de Taubaté, a fin de canjear y evaluar las deficiencias de enfermos de lepra. Se analizaron los registros de 36 nuevos casos, entre enero de 2003 y julio de 2004. Se identificaron 92 contactos, sendo 64,1% deficiencias, tendo 25% de 20 a 29 años y 58,5% mujeres. De estos, 61% fueron sometidos a consulta en la casa de los enfermos . Informó de la no pertenencia al control en el olvido (66,6%, y la falta de tiempo (11,1%, 4 contactos había síntomas de la lepra, y se confirmó 1 (formulario transmisibles - Dimorfa. Sin lograr una cobertura del 100% del activo de búsqueda, le sugerimos que, para garantizar el objetivo de eliminar la enfermedad, las asociaciones y la descentralización de las actividades de control.Descriptive study, with qualitative variables, carried on in the Ambulatory of Specialties Region of Taubaté, in order to evaluate the missing contacts of leprosy patients. Thirty-six records of new cases were analyzed, from January 2003 to July 2004. Ninety-two contacts were identified, being 64.1% missing ones, having 25% from 20 to 29 years

  2. Análise da freqüência de recaídas de malária por Plasmodium vivax em região não endêmica (São Paulo, Brasil Analysis of the frequency of relapses due to malaria caused by Plasmodium vivax in a non endemie area (São Paulo, Brazil

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    Marcos Boulos

    1991-04-01

    Full Text Available Em virtude da existência de poucas informações, devidamente registradas, sobre freqüência e épocas de recaídas de malária por Plasmodium vivax, contraída no Brasil, foi analisada casuística observada em região não endêmica e constituída por pacientes corretamente tratados. O índice de recaídas documentadas em São Paulo, foi alto (24,5%, com desenvolvimento precoce na maioria das oportunidades, ou seja, em tempo inferior a três meses.Very few well-established information is available about the frequency and timeliness of relapses in cases of Plasmodium vivax malaria acquired in Brazil. So, we analysed a series of correctly treated patients observed out of endemic areas. The rate of relapses seen in São Paulo, which may represent that of the parasitosis in the whole country, was high, ranging from 7.5% to 24.5%, and early in most cases, i.e. appearing by three months, what anticipates a high endemicity.

  3. Implication of the E. coli K12 uvrA and recA genes in the repair of 8-methoxypsoralen-induced mono adducts and crosslinks on plasmid DNA; Implicacion de los genes uvrA de E. coli K12 en la reparacion de monoaductos y entrecruzamien tos inducidos en DNA plasmidico por 8-metoxipso raleno mas luz ultravioleta A

    Energy Technology Data Exchange (ETDEWEB)

    Paramio, J.M.; Bauluz, C.; Vidania, R. de

    1986-07-01

    Genotoxicity of psoralen damages on plasmid DNA has been studied. pBR322 DNA was randomly modified with several concentrations of 8-methoxypsoralen plus 365 nm-UV light. After transformation into E. coli strains (wild-type, uvrA and recA) plasmid survival and mutagenesis were analyzed. To study the influence of the SOS response on plasmid recovery, preirradiation of the cells was performed. In absence of cell preirradiation, crosslinks were not repaired in any strain. Mono adducts were also lethal but in part removed by the excision-repair pathway. Preirradiation of the cells significantly. increased plasmid recovery in recA+ celia. In uvrA- only the mutagenic pathway seemed to be involved in the repair of the damaged DNA. Wild type strain showed the highest increase in plasmid survival, involving the repair of mono adducts and some fraction of crosslinks mainly through an error-free repair pathway. This suggests an enhancement of the excision repair promoted by the induction of SOS functions. (Author) 32 refs.

  4. Protocolo POG 9061 en la recaída aislada a sistema nervioso central en pacientes con diagnóstico de leucemia linfoide aguda. Resultados de una serie de casos en el Hospital Universitario de Santander POG 9061 protocol to treat isolated central nervous system recurrence in patients with acute lymphoid leukemia at Hospital Universitario de Santander

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    Ernesto Rueda

    2010-04-01

    Full Text Available Introducción: La leucemia linfoide aguda (LLA es la neoplasia más común en niños; el 5-10 % presentan recaídas a sistema nervioso central (SNC, un factor de mal pronóstico. Objetivo: Describir los resultados obtenidos en una unidad de oncología pediátrica con el protocolo POG 9061 modificado, en pacientes con LLA y recaída aislada a SNC. Metodología: Cohorte de los pacientes atendidos en el Hospital Universitario de Santander (HUS. Se estimó la sobrevida libre de evento (SLE y la sobrevida total (ST, así como las alteraciones resultantes del protocolo. Se incluyeron 15 pacientes atendidos entre enero/93 y marzo/07; el último diagnóstico de recaída al SNC se hizo en noviembre/04. Resultados: El 66,6 % de las recaídas se dieron antes de 18 meses luego de remitir la LLA. Dos pacientes abandonaron el protocolo, uno de los cuales falleció; dos o más fallecieron luego de terminar el protocolo. La ST a cinco años fue de 85,6 % (IC95 % 53,3-96,2, mientras que la SLE de 84,9 % (IC95 % 51,2-96,0 %. La complicación más frecuente fue mielosupresión; no hubo alteraciones de la función renal y solo una ligera elevación de las pruebas de función hepática. Las causas de hospitalización fueron principalmente infecciones. El coeficiente intelectual de los pacientes posterior a la aplicación del protocolo indicaba deficiencia leve en el 45,4 % de ellos. Conclusiones: La sobrevida, el tipo y la frecuencia de complicaciones, son similares a las encontradas a nivel mundial, lo que es relevante dada la alta proporción de pacientes con recaída precoz luego de remisión de la LLA. Salud UIS 2010; 42: 7-17Introduction: Acute lymphoblastic leukemia (ALL is the most frequent neoplasm in children. Relapses to central nervous system (CNS Appears in 5-10 % of the ALL patients and is a bad prognostic factor. Objective: To describe the results obtained with modified POG 9061 protocol in a pediatric oncology unit. Methodology: Survival analysis was

  5. Retraso en el diagnóstico de lepra como factor pronóstico de discapacidad en una cohorte de pacientes en Colombia, 2000 - 2010 Delay in leprosy diagnosis as a predictor of disability in a cohort of patients in Colombia, 2000 - 2010

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    Martha Inírida Guerrero

    2013-02-01

    Full Text Available OBJETIVO: Evaluar los factores pronósticos de la presencia de discapacidad al momento del diagnóstico de lepra en una cohorte de pacientes colombianos de 2000 a 2010. MÉTODOS: Estudio analítico y observacional descriptivo de una cohorte retrospectiva de pacientes ingresados con diagnóstico de lepra en el Centro Dermatológico Federico Lleras Acosta, de Bogotá, Colombia, entre 2000 y 2010. Se realizó el análisis descriptivo de las variables y se identificaron factores pronósticos de la presencia de discapacidad al momento del diagnóstico mediante análisis simple y multifactorial (modelo de riesgos proporcionales de Cox; se calcularon las razones de riesgo (hazard ratio para cada uno de los factores incluidos en el modelo. RESULTADOS: El tiempo entre los primeros síntomas y el diagnóstico en los 333 pacientes de la cohorte fue en promedio 2,9 años; 32,3% de ellos tenían algún grado de discapacidad, especialmente en los pies. Hubo una mayor proporción de retraso en el diagnóstico y discapacidad en hombres que en mujeres y en pacientes con lepra multibacilar que con paucibacilar. La discapacidad se asoció significativamente con demoras ≥ 1 año en el diagnóstico, edad ≥ 30 años, índice baciloscópico inicial ≥ 2, lepra multibacilar y proceder de Cundinamarca o Santander. Los factores protectores fueron ser del sexo femenino, tener algún grado de escolaridad y residir en Boyacá. CONCLUSIONES: El tiempo entre los primeros síntomas y el diagnóstico constituye el factor pronóstico clave de la discapacidad al momento del diagnóstico de lepra. Se recomienda reforzar la búsqueda activa de personas infectadas y promover el diagnóstico precoz.OBJECTIVE: Evaluate predictive factors of disability at time of leprosy diagnosis in a cohort of Colombian patients, from 2000 to 2010. METHODS: Descriptive and analytical observational study of a retrospective cohort of patients admitted with a leprosy diagnosis to the Centro

  6. Adolpho Lutz and controversies over the transmission of leprosy by mosquitoes Adolpho Lutz e as controvérsias sobre a transmissão da lepra por mosquitos

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    Jaime L. Benchimol

    2003-01-01

    -sucedidas estas pesquisas, sustentou a transmissão da lepra por mosquitos nas comissões e congressos de que participou, até sua morte em outubro de 1940.

  7. Revisão sobre o uso da terapia cognitiva-comportamental na prevenção de recaídas e recorrências depressivas: a review Cognitive-behavioral therapy in prevention of depression relapses and recurrences

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    Alexander Moreira de Almeida

    2003-10-01

    Full Text Available OBJETIVO: Fazer um levantamento das teorias e revisar as evidências existentes sobre o papel da terapia cognitiva-comportamental (TCC na prevenção de recaídas e recorrências de episódios depressivos. MÉTODO: Revisão dos ensaios clínicos randomizados e controlados que investigam a eficácia da TCC na prevenção de recaídas e/ou recorrências depressivas. As bases de dados consultadas foram o Medline, Lilacs, Cochrane, Biosis e a Embase. Foram verificadas as referências bibliográficas dos artigos selecionados, de artigos de revisão e de livros sobre o tema. RESULTADOS: Foram encontrados 15 estudos de desenhos heterogêneos e vários deles com problemas metodológicos. A maioria comparou o uso de TCC e antidepressivos apenas na fase aguda. Em 12 deles a TCC diminuiu a taxa de recorrência/recaídas de modo estatisticamente significativo. As publicações mais recentes apontam para a utilidade da TCC nos sintomas depressivos residuais como estratégia preventiva de recorrências. CONCLUSÕES: A TCC mostrou-se eficaz na redução de recaídas depressivas, mas ainda precisam ser mais bem investigadas sua eficácia em relação ao uso de antidepressivos e qual estratégia de TCC (seu uso apenas na fase aguda, na fase aguda e manutenção, na manutenção após antidepressivo na fase aguda ou o enfoque nos sintomas residuais após antidepressivo na fase aguda é mais eficaz para cada tipo de paciente.OBJECTIVES: To revise theories and the available evidence about Cognitive Behavioral Therapy (CBT role on the relapse and recurrence prevention of depressive episodes. METHODS: Review of random and controlled clinical trials that investigated CBT efficacy on the relapse and recurrence prevention of depressive episodes. The following databases were used: Medline, Lilacs, Cochrane, Biosis and Embase. The reference sections of the selected articles, review articles and specialized books were consulted. RESULTS: Fifteen studies with different

  8. Lepra--stadig aktuel i Danmark

    DEFF Research Database (Denmark)

    Jensen, A G; Bygbjerg, Ib Christian; Jensen, H

    1992-01-01

    A case of borderline tuberculoid leprosy in a 27 year old woman from the Philippines is presented. The diagnosis was made after repeated biopsies. Only a single mycobacterium was present and the histology of the initial biopsies was inconclusive. This case emphasizes that leprosy is still imported...

  9. Predictors of relapse in the second follow-up year post cognitive-behavior therapy for panic disorder Preditores de recaída no segundo ano após terapia cognitivo-comportamental para pacientes com transtorno de pânico

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    Elizeth Heldt

    2011-03-01

    Full Text Available OBJECTIVE: To investigate predictors of relapse two years after a brief cognitive-behavior group therapy in patients with panic disorder who had failed to respond to pharmacologic treatment. METHOD: A total of 56 patients with panic disorder were followed who had met remission criteria at 1 year evaluation after 12 sessions of cognitive-behavior group therapy. Demographic and clinical features and life stressors were investigated as predictors of relapse. RESULTS: At the 2 year assessment, 39 (70% patients maintained remission status and use of medication was reduced significantly, such that 36 (64% patients were not undergoing any psychiatric treatment. Among all independent variables investigated, only "conflict" as a stressful life event, RR = 3.20 (CI95% 1.60; 7.20 - p = 0.001, and the severity or residual anxiety symptoms, RR = 3.60 for each scale point (CI95% 1.02; 1.08 - p OBJETIVO: Investigar os preditores de recaída após dois anos de terapia cognitivo-comportamental em grupo breve para pacientes com transtorno do pânico que não responderam ao tratamento farmacológico. MÉTODO: Um total de 56 pacientes com transtorno do pânico que preencheram os critérios de remissão em um ano de avaliação após as 12 sessões da terapia cognitivo-comportamental em grupo foram acompanhados. As características demográficas, clínicas e os estressores de vida foram investigados como preditores de recaída. RESULTADOS: No segundo ano de avaliação, 39 (70% pacientes mantiveram-se em remissão e o uso de medicação reduziu significativamente, de tal forma que 36 (64% pacientes não estavam em tratamento psiquiátrico. Entre todas as variáveis independentes investigadas, somente o "conflito" como evento estressor de vida, RR = 3,20 (CI95% 1,60; 7,20 - p = 0,001 e a gravidade ou os sintomas residuais de ansiedade, RR = 3,60 para cada ponto a mais da escala (CI95% 1,02; 1,08 - p < 0,001, foram preditores de recaída. CONCLUSÃO: A despeito dos

  10. A profile of patients treated at a national leprosy outpatient referral clinic in Rio de Janeiro, Brazil, 1986-2007 Perfil de los enfermos tratados en un servicio nacional de remisión de pacientes ambulatorios con lepra en Río de Janeiro, Brasil, 1986-2007

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    Mariana A. Hacker

    2012-06-01

    Full Text Available OBJECTIVE: To analyze a profile of patients treated at a national leprosy outpatient referral clinic in metropolitan Rio de Janeiro, Brazil, over a period of more than two decades, and the subgroup of nationally registered leprosy cases from the same residential area, as well as all registered cases statewide. METHODS: An observational, descriptive analysis was carried out for patients treated from 1986 to 2007 at the Souza Araújo Outpatient Clinic (Ambulatório Souza Araújo, ASA, a national referral center for the diagnosis and treatment of leprosy at the Oswaldo Cruz Foundation (Fiocruz that serves clients from the city of Rio de Janeiro and other municipalities in the metropolitan area of Rio de Janeiro State. Demographic and clinical data for the subgroup of leprosy cases registered with Brazil's National Disease Notification System (Sistema Nacional de Informação de Agravos de Notificação, SINAN between 2001 and 2007 and residing in the same municipalities as the ASA patients, and for all registered cases statewide, were also analyzed. RESULTS: Among the ASA patients, there was a decrease in average family income (from 3.9 to 2.7 times the minimum salary between the periods 1998-2002 and 2003-2007; the proportion of multibacillary (MB patients (from 52.7% to 46.9%; and the proportion of patients younger than 15 years old (from 12.8% to 8.7%. Among the MB patients, the average initial and final bacilloscopic indices were significantly higher in 2003-2007. Compared with the SINAN cases, more ASA cases involved disability and were younger than 15 years old. CONCLUSIONS: Patients living with leprosy in the metropolitan area of the state of Rio de Janeiro belong to the most deprived social strata and have not benefited from the overall improvement in socioeconomic conditions in Brazil.OBJETIVO: Analizar el perfil de los enfermos tratados en un servicio nacional de remisión de pacientes ambulatorios con lepra ubicado en la zona

  11. Spatial analysis of leprosy incidence and associated socioeconomic factors Análisis espacial de la incidencia de lepra y factores socioeconómicos asociados Análise espacial da incidência de hanseníase e fatores socioeconômicos associados

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    Maria Rita de Cassia Oliveira Cury

    2012-02-01

    Full Text Available OBJECTIVE: To identify clusters of the major occurrences of leprosy and their associated socioeconomic and demographic factors. METHODS: Cases of leprosy that occurred between 1998 and 2007 in São José do Rio Preto (southeastern Brazil were geocodified and the incidence rates were calculated by census tract. A socioeconomic classification score was obtained using principal component analysis of socioeconomic variables. Thematic maps to visualize the spatial distribution of the incidence of leprosy with respect to socioeconomic levels and demographic density were constructed using geostatistics. RESULTS: While the incidence rate for the entire city was 10.4 cases per 100,000 inhabitants annually between 1998 and 2007, the incidence rates of individual census tracts were heterogeneous, with values that ranged from 0 to 26.9 cases per 100,000 inhabitants per year. Areas with a high leprosy incidence were associated with lower socioeconomic levels. There were identified clusters of leprosy cases, however there was no association between disease incidence and demographic density. There was a disparity between the places where the majority of ill people lived and the location of healthcare services. CONCLUSIONS: The spatial analysis techniques utilized identified the poorer neighborhoods of the city as the areas with the highest risk for the disease. These data show that health departments must prioritize politico-administrative policies to minimize the effects of social inequality and improve the standards of living, hygiene, and education of the population in order to reduce the incidence of leprosy.OBJETIVO: Identificar áreas con mayor ocurrencia de casos de lepra y factores socioeconómicos y demográficos asociados. MÉTODOS: Casos de lepra ocurridos entre 1998 y 2007 en Sao José do Rio Preto, Sureste de Brasil, fueron geocodificados y se calcularon los coeficientes de incidencia por sectores censados. Por medio de la técnica de análisis de

  12. Full remission and relapse of obsessive-compulsive symptoms after cognitive-behavioral group therapy: a two-year follow-up Remissão completa e recaídas dos sintomas obsessivo-compulsivos depois da terapia cognitivo-comportamental em grupo: dois anos de acompanhamento

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    Daniela Tusi Braga

    2010-06-01

    Full Text Available OBJECTIVE: The aim of this study was to assess whether the results obtained with 12 sessions of cognitive-behavioral group therapy with obsessive-compulsive patients were maintained after two years, and whether the degree of symptom remission was associated with relapse. METHOD: Forty-two patients were followed. The severity of symptoms was measured at the end of cognitive-behavioral group therapy and at 18 and 24 months of follow-up. The assessment scales used were the Yale-Brown Obsessive-Compulsive Scale, Clinical Global Impression, Beck Depression Inventory, and Beck Anxiety Inventory. RESULTS: The reduction in symptom severity observed at the end of treatment was maintained during the two-year follow-up period (F = 57.881; p OBJETIVO: Avaliar se os resultados obtidos com 12 sessões de terapia cognitivo-comportamental em grupo para pacientes com transtorno obsessivo-compulsivo foram mantidos depois de dois anos do final do tratamento e se o grau de remissão dos sintomas esteve associado às recaídas. MÉTODO: Quarenta e dois pacientes foram acompanhados. A gravidade dos sintomas foi avaliada no final da terapia cognitivo-comportamental em grupo, 18 e 24 meses após o término do tratamento. As escalas utilizadas para avaliação foram Yale-Brown Obsessive-Compulsive Scale, Clinical Global Impression, Beck Depression Inventory e Beck Anxiety Inventory. RESULTADOS: A redução da gravidade dos sintomas observada no final do tratamento foi mantida durante o período de dois anos de acompanhamento (F = 57,881; p < 0,001. Ao final do tratamento, 9 (21,4% pacientes apresentaram remissão completa, 22 (52,4% remissão parcial e 11 (26,2% não apresentaram mudança na Yale-Brown Obsessive-Compulsive Scale. Dois anos depois, 13 pacientes (31,0% apresentaram remissão completa dos sintomas, 20 (47,6% apresentaram remissão parcial, e 9 (21,4% não apresentaram mudança na Yale-Brown Obsessive-Compulsive Scale. A remissão completa dos sintomas ao

  13. Ter hanseníase e trabalhar na enfermagem: história de lutas e superação Tener la lepra y trabajar como enfermera: historias de luchas y superación To be with leprosy and to work as a nurse: histories of fight and overcoming

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    Ana Paula Batista Gusmão

    2009-12-01

    Full Text Available A pesquisa teve como objetivo conhecer a história dos ex-trabalhadores de enfermagem, que tiveram hanseníase internados compulsoriamente desde início do século XX no Brasil. A hanseníase, permeada de preconceitos e estigmas, no início dos anos 20 foi tratada por décadas com severas políticas públicas de isolamento compulsório dos doentes em Hospitais Colônias. Devido ao preconceito, havia dificuldade em contratar recursos humanos para manter estes estabelecimentos. As histórias dos sujeitos pesquisados surgiram a partir de entrevistas gravadas com sete moradores da colônia, pelo método da história oral. Os resultados da pesquisa apresentados parcialmente neste trabalho relatam as histórias de vida dos sujeitos, suas lutas, sofrimentos e sonhos. Confirmou-se que os próprios doentes eram mantenedores dos serviços dentro das colônias, inclusive os de Enfermagem.Esta investigación tuvo como objetivo conocer la historia de los ex-trabajadores, que tuvieron la lepra admitidos en carácter obligatorio a partir del siglo XX en Brasil. La lepra, cargada de prejuicios y el estigma en los 20 años fue tratado durante décadas con las políticas estrictas de aislamiento forzoso de los pacientes en los hospitales colonias. Debido a los prejuicios, hubo dificultad en la contratación de personal para mantener estos establecimientos. Las historias de las personas fueran obtenidas por medio de entrevistas con siete residentes de la colonia, por el método de la historia oral. Los resultados de la investigación en parte se presentan en este documento se informaba de las historias de vida de las personas, sus luchas, penas y sueños. Se confirmó que los mismos pacientes fueron los poseedores de los servicios en las colonias, incluida la enfermería.This research aimed at knowing the history of ex-workers, leprosy patients admitted compulsorily from the early twentieth century in Brazil. Leprosy, fraught with prejudice and stigma in the early

  14. Socioeconomic and demographic profile of leprosy carriers attended in nursing consultations Perfil socioeconómico y demográfico de portadores de lepra atendidos en consulta de enfermería Perfil socioeconômico e demográfico de portadores de hanseníase atendidos em consulta de enfermagem

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    Marli Teresinha Cassamassimo Duarte

    2007-10-01

    Full Text Available Leprosy is a contagious infectious disease that manifests due to unfavorable socioeconomic factors, endemic levels and individual conditions. This study aimed to recognize the socioeconomic and demographic profile and degree of incapacity installed in leprosy carriers attended at the School Health Center in Botucatu. Data were obtained through nursing consultation performed in 37 patients. The results showed a predominance of individuals with stable union (78%, whites (92%, age between 30 and 49 years old (51%, low level of schooling (68% with incomplete primary education and those with per capita familial income less than one minimum salary (59 %. More than one third of the patients investigated (35% presented some degree of physical incapacity. The association of low socioeconomic profile with the presence of physical incapacities assigns greater vulnerability to this population and can negatively influence their quality of life.La lepra es una enfermedad infecto-contagiosa que se manifiesta debido a factores socioeconómicos desfavorables, niveles de endemia y condiciones individuales. El objetivo de este estudio fue reconocer el perfil socioeconómico y demográfico y el grado de incapacidad instalado de los portadores de lepra atendidos en el Centro de Salud Escuela de Botucatu. Los datos fueron obtenidos a través de consulta de enfermería realizada en 37 pacientes. Los resultados mostraron una predominancia de individuos con unión estable (97,8%, blancos (92%, con edad entre 30 y 49 años (51%, con bajo nivel de escolaridad (68 % con enseñanza primaria incompleta y con renta familiar per capita menor que un salario mínimo (59 %. Más de un tercio de los pacientes investigados (35% presentaban algún grado de incapacidad física. La asociación del bajo perfil socioeconómico con la presencia de incapacidades físicas imprime mayor vulnerabilidad a esa población, lo que puede influenciar negativamente su calidad de vida.A hansen

  15. Relación del polimorfismo TaqI del gen del receptor de la vitamina D con la lepra lepromatosa en población mexicana Association between the TaqI polymorphism of Vitamin D Receptor gene and lepromatous leprosy in a Mexican population sample

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    Jesús Salvador Velarde Félix

    2009-02-01

    Full Text Available OBJETIVO: Determinar la relación del polimorfismo TaqI del gen del receptor de la vitamina D (RVD con la lepra lepromatosa (LL en individuos originarios de Sinaloa, México. MATERIAL Y MÉTODOS: Se amplificó un fragmento de 740 pb del gen RVD en muestras de ADN de 71 pacientes con LL y 144 controles en el Hospital General de Culiacán durante el periodo 2004-2007. El polimorfismo se identificó mediante la endonucleasa TaqI. RESULTADOS: Se observó un aumento de relevancia estadística del genotipo TT en pacientes con LL en comparación con los controles (p= 0.040; RM= 1.82. CONCLUSIÓN: Se demuestra un nexo entre el genotipo TT y la susceptibilidad a la LL.OBJETIVE: To establish the association of the vitamin D receptor gene TaqI polymorphism with lepromatous leprosy (LL in individuals from Sinaloa, Mexico. MATERIAL AND METHODS: A 740 bp fragment was amplified from the VDR gene in DNA samples of 71 patients with LL and 144 controls in the Hospital General de Culiacán during 2004-2007. Polymorphism was identified through TaqI endonuclease. RESULTS: A significant increase in the genotype TT of the VDR gene was observed in patients when compared to controls (p = 0.040; OR = 1.82. CONCLUSIONS: Our data support the association between the TT genotype and susceptibility to LL in this Mexican population.

  16. Target Molecular Simulations of RecA Family Protein Filaments

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    Yeng-Tseng Wang

    2012-06-01

    Full Text Available Modeling of the RadA family mechanism is crucial to understanding the DNA SOS repair process. In a 2007 report, the archaeal RadA proteins function as rotary motors (linker region: I71-K88 such as shown in Figure 1. Molecular simulations approaches help to shed further light onto this phenomenon. We find 11 rotary residues (R72, T75-K81, M84, V86 and K87 and five zero rotary residues (I71, K74, E82, R83 and K88 in the simulations. Inclusion of our simulations may help to understand the RadA family mechanism.

  17. Políticas públicas na Hanseníase: contribuição na redução da exclusão social La política pública en la Lepra: contribución en la reducción de la exclusión social Public policies in Leprosy: contribution in reduction of social exclusion

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    Regina Célia Dias

    2008-11-01

    Full Text Available O estudo analisa o desempenho do setor de hanseníase de um município do estado de Minas Gerais, no período de janeiro de 1996 a dezembro de 2004, por meio de um estudo descritivo e retrospectivo dos prontuários dos pacientes que iniciaram poliquimioterapia neste período. Verificou-se a freqüente presença da supervisão técnica estadual, com atendimento aos casos mais graves, cursos de capacitação aos profissionais e o constante apoio frente às dificuldades. O resultado foi o aumento do número de casos detectados e a melhoria na qualidade do atendimento ao portador de hanseníase. Atualmente, 100% dos casos são avaliados quanto ao grau de incapacidade, além do monitoramento das reações hansênicas e neurites, prevenindo assim as deformidades.El estudio examina el desempeño del sector de la lepra de un municipio de estado de Minas Gerais, a partir de enero de 1996 a diciembre de 2004, a través de un estudio descriptivo y retrospectivo de historias clínicas de los pacientes que iniciaron poli quimioterapia en este período. Fue verificada a frecuente presencia de la supervisión técnica del Estado, con atención a los casos más graves, cursos de formación para los profesionales y apoyo constante ante las dificultades. El resultado fue el aumento del número de casos detectados y la mejora en la calidad de la atención al portador de la lepra. Actualmente, el 100% de los casos son evaluados por el grado de discapacidad, además de supervisar las reacciones y neuritis, evitando así deformaciones.The study examines the performance of the sector of leprosy as a municipality of Minas Gerais, from January 1996 to December 2004, through a descriptive and retrospective study of medical records of patients who started multi-drug therapy in this period. It was the frequent presence of the state technical supervision, with attention to the most severe cases, courses of training for professionals and constant support in the face of

  18. Biosynthesis of the Cyclotide MCoTI-II using an Engineered Intein

    Energy Technology Data Exchange (ETDEWEB)

    Cantor, J; Camarero, J A

    2006-08-15

    Cyclotides are an emerging family of naturally occurring circular mini-proteins ({approx}30-40 amino acids) characterized by six conserved Cys residues (forming 3 disulfide bridges) that create a topologically unique structure designated as a cyclic cysteine knot (CCK). The cysteine knot motif, which is embedded within the macrocylic backbone, is described as two disulfide bridges that form a ring that is penetrated by the third disulfide bridge. The cyclic backbone and CCK motif together confer cyclotides with a remarkable stability and resistance to proteolytic, chemical, and thermal degradation. Further, cyclotides are functionally diverse and display a wide range of functions including uterotonic activity, trypsin inhibition, cytotoxicity, neurotensin binding, anti-HIV, antimicrobial, and insecticidal activity. Together, these characteristics make cyclotides attractive candidates for both drug design and agricultural applications, both in their native forms and as molecular scaffolds for the incorporation of novel bioactivities. [1] The ability to manipulate production of cyclotides within biological systems is critical for mutagenesis studies, production of grafted products, and the mass production of cyclotides with novel activities. My adviser's hope is to achieve this capability by employing recombinant DNA expression techniques to generate large combinatorial libraries of cyclotides. The advantage in creating a biosynthetic library (containing {approx}10{sup 6}-10{sup 10} members/library vs. chemically based libraries with typical values ranging from {approx}10{sup 3}-10{sup 5} members/library) is that it can be lead to the in vivo application of biological screening and selection methodologies based on a specific clone's ability to affect certain cellular processes.

  19. Atenção básica de saúde e a assistência em Hanseníase em serviços de saúde de um município do Estado de São Paulo La atención básica a la salud y la asistencia a la Lepra en servicios de salud de un municipio del Estado de São Paulo The basic health and assistance to Hansen's Disease in health care services of a municipality of São Paulo State

    Directory of Open Access Journals (Sweden)

    Adriana Jimenez Pereira

    2008-11-01

    Full Text Available Este é um estudo descritivo desenvolvido em um município do Estado de São Paulo. Objetivo: identificar e caracterizar as ações do Programa de Controle da Hanseníase nos serviços de saúde municipais. Metodologia: entrevistas gravadas com gestor municipal de saúde e profissionais da assistência à hanseníase. Resultados: a política pública municipal em saúde prioriza o desenvolvimento da atenção básica com ênfase na saúde pública. As ações são realizadas por profissionais capacitados e experientes em hanseníase. Ve rificou-se a não realização da busca ativa dos casos, necessária para o real conhecimento da situação epidemiológica, e das ações de educação em saúde, importante para a redução do estigma e aproximação do sujeito à nova situação de vida e enfrentamento de limitações.Este estudio descriptivo es una investigación que analizó la situación de la atención de la Lepra en un municipio del Estado de Sao Paulo. Objetivo: identificar y caracterizar las acciones del Programa de Control de Lepra de los servicios de salud de ese municipio. Metodología: se entrevistaron a los profesionales encargados de la atención de lepra y al director municipal de políticas de salud. Resultados: las políticas públicas municipales de salud priorizaron el desarrollo de la atención básica, con énfasis en la salud pública tradicional. Las acciones de control de lepra son realizadas por trabajadores capacitados y con significativa experiencia profesional. Se resalta la ausencia de busca activa de los casos, necesaria para un conocimiento real de la situación epidemiológica y la importancia de educación en salud, para reducir el estigma y aproximar el sujeto a las adaptaciones necesarias en la nueva situación de vida para afrontar las limitaciones.This descriptive study was carried out in a municipality of Sao Paulo State. The objective was to identify and to characterize the Leprosy Control Program in

  20. Hanseníase: estigma e preconceito vivenciados por pacientes institucionalizados em Santa Catarina (1940-1960 Lepra: estigma y prejuicio vivido por pacientes institucionalizados en Santa Catarina, Brasil (1940-1960 Leprosy: stigma and prejudice lived by institucionalized patients in Santa Catarina State, Brazil (1940-1960

    Directory of Open Access Journals (Sweden)

    Miriam Süssking Borenstein

    2008-11-01

    Full Text Available Trata-se de uma pesquisa qualitativa com abordagem sócio-histórica cujo objetivo foi conhecer o estigma e preconceito vivenciado por pacientes/residentes institucionalizados acometidos pela hanseníase. Para tanto, foram entrevistados três pacientes que viveram em um hospital colônia no período do estudo, utilizando-se o método de história oral. Os dados foram coletados e posteriormente analisados, utilizando-se do referencial de estigma. Os resultados evidenciaram que após o ingresso na instituição, esses doentes tiveram seus laços familiares rompidos, perderam seus direitos como cidadãos, em decorrência dessa situação, assumiram novas vidas em um novo contexto. Concluindo, o isolamento nosocomial por um longo período de tempo (anos de internação e afastamento, provocou a morte simbólica de muitos pacientes que viviam com a esperança de um retorno ao convívio familiar e/ou social.El presente es un estudio de investigación cualitativa con abordaje socio-histórica. Tuvo como objetivo conocer el estigma y el prejuicio vivido por los pacientes institucionalizados portadores de lepra. Utilizándose el método de história oral, fueran entrevistados tres pacientes, que vivieron en un hospital en el período del estudio. Los datos fueran colectados y después analizados utilizándose el referencial del estigma. Los resultados colocaran en evidencia que, después del ingreso en la instituición, las personas tuvieran sus lazos familiares rotos, perdieran sus derechos como ciudadanos y asumiran nuevas vidas en un nuevo contexto. En conclusión, el aislamiento nosocomial por uno largo período de tiempo (años de internación y aislamiento, provocó la muerte simbólica de muchos que vivian con la esperanza del regreso a la convívencia familial y social.This study is a qualitative research with a socio-historic approach whose objective was to know the prejudice and stigma lived by the institutionalized patients/residents with

  1. Hanseníase no município de Fortaleza, CE, Brasil: aspectos epidemiológicos e operacionais em menores de 15 anos (1995-2006 La Lepra en el municipio de Fortaleza, CE, Brasil: aspectos epidemiológicos y operacionales en menores de 15 años (1995-2006 Hansen's Disease in the municipality of Fortaleza, CE, Brazil: epidemiological and operational aspects in children under 15-years-old (1995-2006

    Directory of Open Access Journals (Sweden)

    Carlos Henrique Morais de Alencar

    2008-11-01

    Full Text Available Estudo transversal que objetiva analisar a ocorrência de casos de hanseníase em menores de 15 anos de idade residentes no Município de Fortaleza e notificados no SINAN. Os indicadores epidemiológicos nesta população mostram hiperendemicidade. Operacionalmente observa-se a manutenção da concentração de atendimento em algumas unidades de referência, apesar de alguns avanços. Diagnóstico tardio, elevado grau de incapacidade no diagnóstico e baixo grau de avaliação de contatos registrados revelam a fragilidade das ações de controle. Ressalta-se a possibilidade de erro diagnóstico frente às características da infecção nesta população. A ocorrência de casos de hanseníase nesta população representa um indicador epidemiológico de grande relevância e sua análise amplia a discussão sobre problemas operacionais na rede de serviços de saúde.Estudio transversal con el objetivo de analizar las ocurrencias de los casos de lepra en menores de 15 años de edad sitiados en la ciudad de Fortaleza y enterados en el SINAN. Los indicadores epidémicos en ésta población se presentaron hiperendémicos. En la operacinalización, se observa el mantenimiento de la concentración de atención en algunas unidades recomendables, a pesar de algunos avances. Diagnósticos muy tardes, gran grado de incapacidad en el diagnóstico y bajo grado de evaluación de contactos registrados, muestran la fragilidad de las acciones de control. Se resalta así la posibilidad de error de diagnóstico delante de las características de la infección en ésta población. La ocurrencia de casos de lepra en ésta población representa un indicador epidémico de gran importancia y su análisis amplia la discusión sobre los problemas operacionales en la red de servicios públicos.This is a sectional study that aims to analyze the occurrence of Hansen's disease cases in 15 year-old minors in the Municipality of Fortaleza and notified in SINAN. The

  2. La lepra y el riñón

    Directory of Open Access Journals (Sweden)

    Gerzain Rodríguez

    1999-03-01

    Full Text Available A review is made of leprosy's effect on the kidneys when related to secondary phenomena, such as precipitation within glomerules of antigen-antibody complexes, or fibrillar proteins resulting from chronic inflamation orto complications dueto polychemotherapy against the leprosy bacillus, ¡.e. not being due to M. Ieprae presence within the renal nerves or parenchyma. The following circumstances have been described: a defects in urine concentration and acidification; b acute and chronic interstitial nephritis; c acute renal failure; d systemic secondary or reactive amyloidosis; and e glomerulonephritis. It is also pointed out that leprosy is not a contraindication for renal transplant, because the disease (having already been cured may not become active again, or, if it does, or erupts because of the irnmunosuppression needed to maintain the transplant, can be controlled with availble polychemotherpy.

  3. Type 2 lepra reactions (ENL presenting with extensive cutaneous ulcerations

    Directory of Open Access Journals (Sweden)

    Sapnashree Budhnoor Sreekantaiah

    2012-01-01

    Full Text Available A 45 year old lady presented with multiple painful necrotic ulcerations over the trunk, arms, thighs and gluteal areas of two months duration. She also had erythematous papules and pustules over the face since 1 week. History of recurrent papular lesions, some of them undergoing ulceration were present since 3 years. All biochemical parameters were within normal limits. Rheumatoid factor, ANA, Elisa for HIV and VDRL were negative. Pus culture showed growth of Staphylococcus aureus. Smear for AFB showed multiple globi. Skin biopsy showed atrophic epidermis with grenz zone; dermis showed sheets of foamy macrophages and oedematous blood vessels infiltrated with neutrophils and ocasional plasma cells. Patient was admitted and was started on MDT-MB along with thalidomide and prednisolone.

  4. Organização de serviços de saúde na eliminação da Hanseníase em municípios do Estado de São Paulo La organización de los servicios de salud en la eliminación de la Lepra en municipios del Estado de São Paulo Health services organization on the elimination of Leprosy in municipalities of São Paulo State

    Directory of Open Access Journals (Sweden)

    Lúcia Maria Frazão Helene

    2008-11-01

    Full Text Available Este trabalho tem como objetivo analisar as ações de controle da hanseníase desenvolvidas em nove municípios do Estado de São Paulo e uma coordenadoria de saúde da Capital, selecionados a partir de sua população, localização geográfica e coeficientes de prevalência. Foi aprovado pelo Comitê de Ética nº 514/05 e foram aplicados instrumentos por meio de entrevistas em 59 profissionais e 17 interlocutores. Os dados foram analisados utilizando o programa SPSS (Statistical Package for Social Scientists. Verificou-se que a prevalência apresenta declínio e a detecção permanece estável. As ações de prevenção de incapacidade física, educação em saúde e a busca ativa são pouco desenvolvidas. Os profissionais mais capacitados encontram-se nas unidades de referência, há fragilidade quanto ao trabalho em equipe.El objetivo de este trabajo fue analizar las acciones de control de la Lepra realizadas en nueve Municipios del Estado de Sao Paulo y en una coordinación de salud de la capital, seleccionados a partir de su población, localización geográfica y coeficientes de prevalencia. Este estudio recibió aprobación del Comité de Ética nº 514/05. Se entrevistaron 59 profesionales y 17 coordinadores de área. Para el análisis de los datos se utilizó el programa SPSS (Statistical Package for Social Scientists. Se verificó un declive en la prevalencia, pero la detección permaneció estable; las acciones de prevención de incapacidad física, educación en salud y la busca activa fueron poco ejecutadas; los profesionales más capacitados pertenecían a las unidades de referencia. Hubo fragilidad en cuanto al trabajo en equipo.The present study aims to analyze actions to control Hansen's disease developed in nine municipalities of the State of São Paulo and one of the health authorities of the city of São Paulo, selected based on their population, geographic location and prevalence coefficients. The study was approved by

  5. Bacillus halodurans RecA-DNA binding and RecA- mediated ...

    African Journals Online (AJOL)

    STORAGESEVER

    2009-05-04

    May 4, 2009 ... is that the external environment and the internal pH of the bacteria are alkaline; here also the bacteria may have .... X-ray data collection, crystals were transferred to the solution above ..... At this pH 7.4 the introduction of inducible mutation in cI101-229DM did not increase the cleavage rate. cI101-.

  6. RECA EXPRESSION IN RESPONSE TO SOLAR UVR IN THE MARINE BACTERIUM VIBRIO NATRIEGENS.

    Science.gov (United States)

    Medicinal plants may carry residuals of environmentally persistent pesticides or assimilate heavy metals in varying degrees. Several factors may influence contaminant accumulation, including species, level and duration of contaminant exposure, and topography. As part of a program...

  7. HuRECA: Human Reliability Evaluator for Computer-based Control Room Actions

    International Nuclear Information System (INIS)

    Kim, Jae Whan; Lee, Seung Jun; Jang, Seung Cheol

    2011-01-01

    As computer-based design features such as computer-based procedures (CBP), soft controls (SCs), and integrated information systems are being adopted in main control rooms (MCR) of nuclear power plants, a human reliability analysis (HRA) method capable of dealing with the effects of these design features on human reliability is needed. From the observations of human factors engineering verification and validation experiments, we have drawn some major important characteristics on operator behaviors and design-related influencing factors (DIFs) from the perspective of human reliability. Firstly, there are new DIFs that should be considered in developing an HRA method for computer-based control rooms including especially CBP and SCs. In the case of the computer-based procedure rather than the paper-based procedure, the structural and managerial elements should be considered as important PSFs in addition to the procedural contents. In the case of the soft controllers, the so-called interface management tasks (or secondary tasks) should be reflected in the assessment of human error probability. Secondly, computer-based control rooms can provide more effective error recovery features than conventional control rooms. Major error recovery features for computer-based control rooms include the automatic logic checking function of the computer-based procedure and the information sharing feature of the general computer-based designs

  8. Consulta de enfermagem ao portador de hanseníase no território da Estratégia da Saúde da Família: percepções de enfermeiro e pacientes Consulta de enfermería al portador de la lepra en lo território de la Estrategia de Salud de la Familia: percepciones de enfermeros y pacientes Nursing consultation for leprosy patients in the territory of the Family Health Strategy: perceptions of nurses and patients

    Directory of Open Access Journals (Sweden)

    Cibelly Aliny Siqueira Lima Freitas

    2008-11-01

    Full Text Available O presente estudo visa identificar as dificuldades do enfermeiro para que ele possa refletir e criar estratégias para melhorar a qualidade da consulta e acompanhamento de enfermagem aos portadores de hanseníase e conhecer a percepção do cliente em relação à estrutura do atendimento e o acompanhamento de enfermagem. A partir dos resultados notamos que na consulta de enfermagem busca-se criar um vínculo de confiança com cliente com objetivo do mesmo receber as informações sobre seu tratamento bem como sobre de incapacidades. No que diz respeito às dificuldades afirmadas pelas enfermeiras, elas estão relacionadas às condições de organização dos serviços de saúde o que acarreta uma alta demanda. Com relação à percepção dos pacientes quanto à consulta de enfermagem mostrou-se, no geral, bem satisfatória.El presente studio objetiva identificar las dificuldades para los enfermeros de modo que ellos puedan refexionar y crear estrategias para mejorar la calidad de la consulta de enfermería y el seguimiento de los pacientes con la Lepra y endtender la percepción del paciente en relación a la esctructura de atención y el seguimiento de enfermería. De los resultados nosotros hemos notado que en la consulta de enfermería se busca la creación de un vínculo de confianza con el paciente con el objetivo de que el reciba las informaciones sobre su tratamiento así cómo sus limitaciones. En lo que tanje a las dificuldades presentadas por las enfermeras, ellas están relacionadas a las condiciones de organización de los servicios de salud lo que produce una alta demanda. Con relación a la percepción de los paciente a la consulta de enfermería, ha sido demonstrado, en general, bien satisfatoria.The present study aims to identify difficulties for the nurse so that he can reflect and create strategies to improve the quality of the nursing consultation and follow-up for patients with leprosy and to understand the perception of

  9. Immunological crossreactivity of the Mycobacterium leprae CFP-10 with its homologue in Mycobacterium tuberculosis

    NARCIS (Netherlands)

    Geluk, A.; van Meijgaarden, K. E.; Franken, K. L. M. C.; Wieles, B.; Arend, S. M.; Faber, W. R.; Naafs, B.; Ottenhoff, T. H. M.

    2004-01-01

    Mycobacterium tuberculosis culture filtrate protein-10 (CFP-10) (Rv3874) is considered a promising antigen for the immunodiagnosis of tuberculosis (TB) together with early secreted antigens of M. tuberculosis (ESAT-6). Both ESAT-6 and CFP-10 are encoded by the RD1 region that is deleted from all

  10. Dependência química e prevenção à “recaída”

    Directory of Open Access Journals (Sweden)

    Fátima Büchele

    2004-01-01

    Full Text Available El presente estudio se trata de una investigación descriptiva-exploratoria con un abordaje cuali-cuantitativo, el cual posibilitó hacer un levantamiento de los factores considerados de riesgo para las reacaidas según la visión de los entrevistados en la Unidad de Dependencia Química del Instituto de Psiquiatría de Santa Catarina. Para la colecta de los datos se aplicó un cuestionario en la forma de entrevista a una muestra de 25 usuarios internados en la misma unidad, durante el período de abril a mayo del 2003. El objetivo fue levantar las situaciones de riesgo que los dependientes químicos reconocen como signos y síntomas siendo que estos los llevan a una recaida. Entre tanto, presentamos los resultados obtenidos donde se demuestran algunos factores de riesgo para la recaida que contribuyen con el área de dependencia química en este sentido.

  11. QUANTIFICATION OF RECA GENE EXPRESSION AS AN INDICATOR OF REPAIR POTENTIAL IN MARINE BACTERIOPLANKTON COMMUNITIES OF ANTARCTICA.

    Science.gov (United States)

    Marine bacteria in surface waters must cope daily with the damaging effects of exposure to solar radiation (containing both UV-A and UV-B wavelengths), which produces lesions in their DNA. As the stratospheric ozone layer is depleted, these coping mechanisms are likely to play an...

  12. Acanthamoeba Sp. S-11 phagocytotic activity on Mycobacterium ...

    African Journals Online (AJOL)

    Background: Mycobacterium leprae (M. leprae) is a pathogenic bacterium that causes leprosy. The presence of M. leprae in the environment is supported by microorganisms that act as the new host for M. leprae. Acanthamoeba's potential to be a host of M. leprae in the environment. Acanthamoeba sp. is Free Living ...

  13. Identification of primary drug resistance to rifampin in Mycobacterium leprae strains from leprosy patients in Amazonas State, Brazil.

    Science.gov (United States)

    Contreras Mejía, Matilde Del Carmen; Porto Dos Santos, Maísa; Villarouco da Silva, George Allan; da Motta Passos, Isabella; Naveca, Felipe Gomes; Souza Cunha, Maria da Graça; Moraes, Milton Ozório; de Paula, Lucia

    2014-12-01

    The aim of this study was to identify polymorphisms in the folp1, gyrA, and rpoB genes in leprosy patients treated in Amazonas State, Brazil. Among 197 slit-skin smear samples from untreated or relapsed patients, we found three cases of primary resistance to rifampin and one confirmed case of multidrug resistance. Copyright © 2014, American Society for Microbiology. All Rights Reserved.

  14. ORF Alignment: NC_004431 [GENIUS II[Archive

    Lifescience Database Archive (English)

    Full Text Available 7:H7] pdb|2REC|F Chain F, Reca ... Hexamer Model, Electron Microscopy pdb|2REC|E Chain E, ... ...Reca Hexamer Model, Electron Microscopy pdb|2REC|D Chain ... D, Reca Hexamer Model, Electron Microscopy... pdb|2REC|C ... Chain C, Reca Hexamer Model, Electron Microscopy ... ... ... pdb|2REC|B Chain B, Reca Hexamer Model, Electron ... Microscopy pdb|2REC|A Chain A, Reca Hexamer ...Model, ... Electron Microscopy ... Length = 326 ... Query: 7 ... KQKALAAALGQIEKQFGKGSIMRLGEDRSMDVET

  15. ORF Alignment: NC_004741 [GENIUS II[Archive

    Lifescience Database Archive (English)

    Full Text Available 7:H7] pdb|2REC|F Chain F, Reca ... Hexamer Model, Electron Microscopy pdb|2REC|E Chain E, ... ...Reca Hexamer Model, Electron Microscopy pdb|2REC|D Chain ... D, Reca Hexamer Model, Electron Microscopy... pdb|2REC|C ... Chain C, Reca Hexamer Model, Electron Microscopy ... ... ... pdb|2REC|B Chain B, Reca Hexamer Model, Electron ... Microscopy pdb|2REC|A Chain A, Reca Hexamer ...Model, ... Electron Microscopy ... Length = 326 ... Query: 7 ... KQKALAAALGQIEKQFGKGSIMRLGEDRSMDVET

  16. ORF Alignment: NC_004337 [GENIUS II[Archive

    Lifescience Database Archive (English)

    Full Text Available 7:H7] pdb|2REC|F Chain F, Reca ... Hexamer Model, Electron Microscopy pdb|2REC|E Chain E, ... ...Reca Hexamer Model, Electron Microscopy pdb|2REC|D Chain ... D, Reca Hexamer Model, Electron Microscopy... pdb|2REC|C ... Chain C, Reca Hexamer Model, Electron Microscopy ... ... ... pdb|2REC|B Chain B, Reca Hexamer Model, Electron ... Microscopy pdb|2REC|A Chain A, Reca Hexamer ...Model, ... Electron Microscopy ... Length = 326 ... Query: 7 ... KQKALAAALGQIEKQFGKGSIMRLGEDRSMDVET

  17. ORF Alignment: NC_000913 [GENIUS II[Archive

    Lifescience Database Archive (English)

    Full Text Available 7:H7] pdb|2REC|F Chain F, Reca ... Hexamer Model, Electron Microscopy pdb|2REC|E Chain E, ... ...Reca Hexamer Model, Electron Microscopy pdb|2REC|D Chain ... D, Reca Hexamer Model, Electron Microscopy... pdb|2REC|C ... Chain C, Reca Hexamer Model, Electron Microscopy ... ... ... pdb|2REC|B Chain B, Reca Hexamer Model, Electron ... Microscopy pdb|2REC|A Chain A, Reca Hexamer ...Model, ... Electron Microscopy ... Length = 326 ... Query: 7 ... KQKALAAALGQIEKQFGKGSIMRLGEDRSMDVET

  18. ORF Alignment: NC_002695 [GENIUS II[Archive

    Lifescience Database Archive (English)

    Full Text Available 7:H7] pdb|2REC|F Chain F, Reca ... Hexamer Model, Electron Microscopy pdb|2REC|E Chain E, ... ...Reca Hexamer Model, Electron Microscopy pdb|2REC|D Chain ... D, Reca Hexamer Model, Electron Microscopy... pdb|2REC|C ... Chain C, Reca Hexamer Model, Electron Microscopy ... ... ... pdb|2REC|B Chain B, Reca Hexamer Model, Electron ... Microscopy pdb|2REC|A Chain A, Reca Hexamer ...Model, ... Electron Microscopy ... Length = 326 ... Query: 7 ... KQKALAAALGQIEKQFGKGSIMRLGEDRSMDVET

  19. PI-PfuI and PI-PfuII, intein-coded homing endonucleases from Pyrococcus furiosus. II. Characterization Of the binding and cleavage abilities by site-directed mutagenesis.

    OpenAIRE

    Komori, K; Ichiyanagi, K; Morikawa, K; Ishino, Y

    1999-01-01

    PI- Pfu I and PI- Pfu II from Pyrococcus furiosus are homing endonucleases, as shown in the accompanying paper. These two endonucleases are produced by protein splicing from the precursor protein including ribonucleotide reductase (RNR). We show here that both enzymes specifically interact with their substrate DNA and distort the DNA strands by 73 degrees and 67 degrees, respectively. They have two copies of the amino acid sequence motif LAGLIDADG, which is present in the majority of homing e...

  20. recA mediated spontaneous deletions of the icaADBC operon of clinical Staphylococcus epidermidis isolates : a new mechanism of phenotypic variations

    NARCIS (Netherlands)

    Nuryastuti, Titik; van der Mei, Henny C.; Busscher, Henk J.; Kuijer, Roel; Aman, Abu T.; Krom, Bastiaan P.

    Phenotypic variation of Staphylococcus epidermidis involving the slime related ica operon results in heterogeneity in surface characteristics of individual bacteria in axenic cultures. Five clinical S. epidermidis isolates demonstrated phenotypic variation, i.e. both black and red colonies on Congo

  1. Eficacia de la terapia grupal, familiar y farmacológica en la prevención de recaídas en alcoholismo

    OpenAIRE

    Ríos Lamas, Paloma

    2017-01-01

    Trabajo de fin de Grado. Grado en Enfermería (Zamora). Curso académico 2016-2017 [ES]El alcoholismo es un trastorno crónico de la conducta. La Organización Mundial de la Salud (OMS) lo define como “una conducta caracterizada por el consumo excesivo, adictivo y dependiente del etanol, a tal grado que afecta a la salud física y mental, sus relaciones y el funcionamiento social y económico.” El objetivo general de este trabajo es evaluar la efectividad de la terapia familiar y grupal, y de...

  2. Single-Molecule Manipulation of Double-Stranded DNA Using Optical Tweezers: Interaction Studies of DNA with RecA and YOYO-1

    NARCIS (Netherlands)

    Bennink, Martin L.; Scharer, Orlando D.; Kanaar, Ronald; Sakata-Sogawa, Kumiko; Schins, J.M.; Kanger, Johannes S.; de Grooth, B.G.; Greve, Jan

    1999-01-01

    By using optical tweezers and a specially designed flow cell with an integrated glass micropipette, we constructed a setup similar to that of Smith et al. (Science 271:795-799, 1996) in which an individual double-stranded DNA (dsDNA) molecule can be captured between two polystyrene beads. The first

  3. UV induction of the LT-Toxin operon with respect to the genes lexA, recA, and umuD

    International Nuclear Information System (INIS)

    Tiganova, I.G.; Rusina, O.Yu.; Andreeva, I.V.; Brukhanskii, G.V.; Skavronskaya, A.G.

    1994-01-01

    UV induction of the elt operon (the LT-toxin operon in Escherichia coli) was demonstrated in experiments using fusion of elt::lac operons with the help of Mud1(Ap lac) phage. UV induction of the elt operon is lexA-dependent; thus, the possibility of SOS regulation of this process may be assumed. However, UV induction of the elt operon turned out to be recA-independent, which makes it impossible to consider this induction as a typical SOS response. UV induction of the elt operon is also observed in Salmonella typhimurium, which differs from E. coli in the product of umuD, which suggests that the UV induction of the elt operon is umuD independent

  4. Life forms employ different repair strategies of repair single- and double strand DNA breaks caused by different qualities of radiation: criticality of RecA mediated repair system

    International Nuclear Information System (INIS)

    Sharan, R.N.

    2013-01-01

    Different qualities of radiation, either through direct or indirect pathway, induce qualitative different spectrum of damages in DNA, which are also different in in vitro and in vivo systems. The single- and double strand breaks of DNA are of special interest as they lead to serious biological consequences. The implications of such damage to DNA and their processing by various inherent repair pathways together decide the fate of the living form

  5. Proliferating cell nuclear antigen (PCNA) interacts with a meiosis-specific RecA homologues, Lim15/Dmc1, but does not stimulate its strand transfer activity

    International Nuclear Information System (INIS)

    Hamada, Fumika N.; Koshiyama, Akiyo; Namekawa, Satoshi H.; Ishii, Satomi; Iwabata, Kazuki; Sugawara, Hiroko; Nara, Takayuki Y.; Sakaguchi, Kengo; Sawado, Tomoyuki

    2007-01-01

    PCNA is a multi-functional protein that is involved in various nuclear events. Here we show that PCNA participates in events occurring during early meiotic prophase. Analysis of protein-protein interactions using surface plasmon resonance indicates that Coprinus cinereus PCNA (CoPCNA) specifically interacts with a meiotic specific RecA-like factor, C. cinereus Lim15/Dmc1 (CoLim15) in vitro. The binding efficiency increases with addition of Mg 2+ ions, while ATP inhibits the interaction. Co-immunoprecipitation experiments indicate that the CoLim15 protein interacts with the CoPCNA protein in vitro and in the cell extracts. Despite the interaction between these two factors, no enhancement of CoLim15-dependent strand transfer activity by CoPCNA was found in vitro. We propose that the interaction between Lim15/Dmc1 and PCNA mediates the recombination-associated DNA synthesis during meiosis

  6. ORF Alignment: NC_006085 [GENIUS II[Archive

    Lifescience Database Archive (English)

    Full Text Available NC_006085 gi|50842495 >1u94A 1 322 7 328 e-122 ... gb|AAT70029.1| RecA [Propionibacterium acne...s] gb|AAT70028.1| RecA ... [Propionibacterium acnes] gb|AAT70027.1| RecA ... [Propionibacterium acne...s] gb|AAT70026.1| RecA ... [Propionibacterium acnes] gb|AAT70025.1| RecA ... [Propionibacterium acne...s] gb|AAT70024.1| RecA ... [Propionibacterium acnes] ...gb|AAT70018.1| RecA ... [Propionibacterium acnes] gb|AAT70014.1| RecA ... [Propionibacterium acne

  7. Rapid cloning and purification of proteins: gateway vectors for protein purification by self-cleaving tags.

    Science.gov (United States)

    Gillies, Alison R; Hsii, Judy F; Oak, Seachol; Wood, David W

    2008-10-01

    We have combined Invitrogen's Gateway cloning technology with self-cleaving purification tags to generate a new system for rapid production of recombinant protein products. To accomplish this, we engineered our previously reported DeltaI-CM cleaving intein to include a Gateway cloning recognition sequence, and demonstrated that the resulting Gateway-competent intein is unaffected. This intein can therefore be used in several previously reported purification methods, while at the same time being compatible with Gateway cloning. We have incorporated this intein into a set of Gateway vectors, which include self-cleaving elastin-like polypeptide (ELP), chitin binding domain (CBD), phasin (polyhydroxybutyrate-binding), or maltose binding domain (MBD) tags. These vectors were verified by Gateway cloning of TEM-1 beta-lactamase and Escherichia coli catalase genes, and the expressed target proteins were purified using the four methods encoded on the vectors. The purification methods were unaffected by replacing the DeltaI-CM intein with the Gateway intein. It was observed that some purification methods were more appropriate for each target than others, suggesting utility of this technology for rapid process identification and optimization. The modular design of the Gateway system and intein purification method suggests that any tag and promoter can be trivially added to this system for the development of additional expression vectors. This technology could greatly facilitate process optimization, allowing several targets and methods to be tested in a high-throughput manner.

  8. “EXPRESIÓN DE LA PROTEINA HSP65 DE Mycobacterium leprae EN TABACO Y SU EFECTIVIDAD COMO TRATAMIENTO ORAL EN RATAS ARTRÍTICAS”

    OpenAIRE

    Rodríguez Narciso, César

    2012-01-01

    Los sistemas de expresión basados en plantas brindan un medio adecuado para producir moléculas inmunogénicas que podrían ser utilizadas como vacunas orales. El tabaco (Nicotiana tabacum), en particular ha sido uno de los mejores sistemas modelo en este campo, ya que diversas investigaciones han demostrado que una amplia variedad de proteínas foráneas pueden ser expresadas eficientemente en esta planta, debido al sistema de transformación bien establecido con que se cuenta. El o...

  9. ORF Alignment: NC_002677 [GENIUS II[Archive

    Lifescience Database Archive (English)

    Full Text Available cobacterium leprae ... Length = 96 ... Query: 52 ... LAGSALLVVKRGPNAGSRFLLDQAITSAGRHPDSDIFLDDVTVSRRHAEFRLEG...NEFHVV 111 ... LAGSALLVVKRGPNAGSRFLLDQAITSAGRHPDSDIFLDDVTVSRRHAEFRLEGNEFHVV Sbjct: 1 ... LAGSALLVVKRGPNAGSRFLLDQAITSAGRHPDSDIFLDDVTVSRRHAEFRLEGNEFHVV 60 ...

  10. ORF Alignment: NC_002677 [GENIUS II[Archive

    Lifescience Database Archive (English)

    Full Text Available in ML0151 [Mycobacterium leprae TN] ... emb|CAC29659.1| conserved hypothetical protein ... [Mycobacterium leprae...] pir||G86927 conserved ... hypothetical protein ML0151 [imported] - Mycobacterium ... leprae... ... Length = 61 ... Query: 27 TDIEELREEIDRLDAQILATVKRRAEVSQAIGK...VRMASGGTRLVHSREMKVIERYSELGP 86 ... TDIEELREEIDRLDAQILATVKRRAEVSQAIGKVRMASGGTRLVHSREMKVIERYSELGP Sbjct: 1 ... TDIEELREEIDRLDAQILATVKRRAEVSQAIGKVRMASGGTRLVHSREMKVIERYSELGP 60 ...

  11. ORF Alignment: NC_002677 [GENIUS II[Archive

    Lifescience Database Archive (English)

    Full Text Available ein ML0802 [Mycobacterium leprae TN] ... emb|CAC30312.1| conserved hypothetical protein ... [Mycobacterium leprae...] pir||D87009 conserved ... hypothetical protein ML0802 [imported] - Mycobacterium ... leprae... ... Length = 69 ... Query: 10 AEDVRAEIVASVLEVVVSEGDQIGKGDVLVLL...ESMKMEIPVLAGVAGIVSKVSVSVGDVI 69 ... AEDVRAEIVASVLEVVVSEGDQIGKGDVLVLLESMKMEIPVLAGVAGIVSKVSVSVGDVI Sbjct: 1 ... AEDVRAEIVASVLEVVVSEGDQIGKGDVLVLLESMKMEIPVLAGVAGIVSKVSVSVGDVI 60 ...

  12. Analysis, Diagnosis and Prognosis of Leprosy Utilizing Fuzzy ...

    African Journals Online (AJOL)

    Leprosy (Hensen's Disease) is an infectious disease that progressively deteriorates over time and is associated with mycobacterium leprae (M.leprae). Although human-to-human transmission is the primary source of infection, three other species can carry and (rarely) transfer M.leprae to humans: chimpanzees, mangabey ...

  13. The interaction of antimicrobial peptides with the membrane and intracellular targets of Staphylococcus aureus investigated by ATP leakage, DNA-binding analysis, and the expression of a LexA-controlled gene, recA

    DEFF Research Database (Denmark)

    Gottschalk, Sanne; Thomsen, Line Elnif

    2017-01-01

    The analysis of how antimicrobial peptides (AMPs) interact with bacterial membranes and intracellular targets is important for our understanding of how these molecules affect bacteria. Increased knowledge may aid the design of AMPs that work on their target bacterium without inducing bacterial re...

  14. Desenvolvimento de um jogo terapêutico para prevenção da recaída e motivação para mudança em jovens usuários de drogas

    Directory of Open Access Journals (Sweden)

    Anna Virginia Williams

    Full Text Available Apesar da alta prevalência do abuso, e dependência de drogas entre jovens, existe uma falta de intervenções originais desenvolvidas especificamente para o tratamento dessa população. Este estudo objetivou desenvolver uma técnica no formato de um jogo de cartas para ser utilizado no tratamento de jovens usuários de drogas. A técnica foi intitulada "Jogo da Escolha", e a sua elaboração envolveu: adaptação da linguagem, avaliação do conteúdo e elaboração de suas instruções junto aos profissionais de dependência química. Após um estudo-piloto, foram realizadas modificações nas instruções e no formato de aplicação, obtendo-se a versão atual do "Jogo da Escolha". O jogo demonstra ser útil para trabalhar crenças típicas de jovens usuários de drogas e promover estratégias de enfrentamento em situações de risco.

  15. Epidemiología del tabaquismo. Efectividad a largo plazo de un programa de deshabituación y estudio de los factores asociados a la recaída.

    OpenAIRE

    Bou Monterde, Ricardo

    2001-01-01

    El tabaquismo es la principal causa de morbimortalidad evitable en todo el mundo desarrollado y en los países en vías de desarrollo, y constituye una verdadera epidemia de nuestro tiempo (Warner, 1993). Tanto en Europa, como en otras partes del mundo, desde hace tiempo se vienen llevando a cabo programas de lucha antitabáquica, con el objetivo de reducir la prevalencia de éste hábito y disminuir sus consecuencias sobre la salud de la población.El presente estudio trata de describir la inciden...

  16. Manejo de recaída en enfermedad de Devic. Primer tratamiento exitoso, reportado con plasmaféresis en Ecuador; revisión bibliográfica, actualidad y reporte de caso

    OpenAIRE

    Alemán-Iñiguez, Juan Miguel; Alemán-Iñiguez, Verónica Alexandra; Díaz-Heredia, Fabián

    2015-01-01

    La enfermedad de Devic es una rara patología desmielinizante que afecta al nervio óptico y que se acompaña de sintomatología medular. A continuación se presenta el éxito terapéutico de la plasmaféresis en este caso, como el primero en Ecuador; asimismo caracterizamos el cuadro y analizamos el beneficio del tratamiento en la historia natural de la enfermedad. Se trata de un paciente masculino, adulto, con antecedente de amaurosis súbita, y que comienza con una mielopatía aguda. En ausencia de ...

  17. ORF Sequence: NC_002677 [GENIUS II[Archive

    Lifescience Database Archive (English)

    Full Text Available NC_002677 gi|15827448 >gi|15827448|ref|NP_301711.1| hypothetical protein ML0938 [Mycobacterium leprae... TN] MEHFLDVSTEPRWTEAQRLVTLAAIDIVKTFFKADRDAAGAKINEYLRRIGSEGVIAGYNGLCGMLVKHVAEVTGEDPMAVVYRAEETARNVLPEEEE

  18. ORF Sequence: NC_002677 [GENIUS II[Archive

    Lifescience Database Archive (English)

    Full Text Available NC_002677 gi|15827020 >gi|15827020|ref|NP_301283.1| putative GTP cyclohydrolase I [Mycobacterium leprae... TN] MALLDLGLESTAVPRIRVFDQQRAEAAIRELLYAIGEDPDREGLADTPARVARACRELFSGLYTDPQTVLNTMFDEEHNELV...IVKEIPMYSTCEHHLVSFHGVAHIGYLPGADGRVTGLSKIARLVDLYAKRPQVQERLTSQIADALVSKLDPRGVIIVVEAEHLCMAMRGVRKPGAITTTSAVRGQFKTDAASRAEALGLILRK

  19. ORF Sequence: NC_002677 [GENIUS II[Archive

    Lifescience Database Archive (English)

    Full Text Available NC_002677 gi|15828228 >gi|15828228|ref|NP_302491.1| putative membrane protein [Mycobacterium leprae... TN] MSKGDRKNGVPSTLTTIPLVDPHAEPTEPSIGDLIKDATTQVSTLVRAEVELARAEIIRDVKKGLTGSVFFIAALVVLFYSTFFFF

  20. ORF Sequence: NC_002677 [GENIUS II[Archive

    Lifescience Database Archive (English)

    Full Text Available NC_002677 gi|15826974 >gi|15826974|ref|NP_301237.1| hypothetical protein ML0151 [Mycobacterium leprae... TN] MRPEPPHHENAELTEMNTEVVEAPLLTDIEELREEIDRLDAQILATVKRRAEVSQAIGKVRMASGGTRLVHSREMKVIERYSELGPDGKDLAILLLRLGRGRLGH

  1. ORF Sequence: NC_002677 [GENIUS II[Archive

    Lifescience Database Archive (English)

    Full Text Available NC_002677 gi|15828329 >gi|15828329|ref|NP_302592.1| possible hemagglutinin. [Mycobacterium leprae... TN] MAENPNVDDLRAPLLAALGAADLALTTVNELVGNMRERAEETRIDTRSRVEESRARVAKLQEVLPEHLSELREKFTADELRKAAEGYL...EAATNRYNELVERGEAALERLRSRPVFEDASARAEGYVDQAVELTQEALGTVASQTRAVGGRAAKLVGIELPKKAAAPARKAPAKKAPAKKAPAKKVTQK

  2. ORF Sequence: NC_002677 [GENIUS II[Archive

    Lifescience Database Archive (English)

    Full Text Available NC_002677 gi|15827196 >gi|15827196|ref|NP_301459.1| putative integration host factor [Mycobacterium leprae... TN] MALPQLTDEQRAAALEKAAAARRARAELKDRLKRGGTNLTQVLKDAESDEVLGKMKVSALLEALPKVGKVKAQEIMTELDIAPTRRLRGLGERQRKALLEKFGSA

  3. ORF Sequence: NC_002677 [GENIUS II[Archive

    Lifescience Database Archive (English)

    Full Text Available NC_002677 gi|15827491 >gi|15827491|ref|NP_301754.1| hypothetical protein ML1018 [Mycobacterium leprae... TN] MAYDDPFMQTESTYWPTLPAGAQQYTSSTKHPVDGDRAEATVDMAGIPCRIMPQRHRNEQHSVAGYASSHR

  4. ORF Sequence: NC_002677 [GENIUS II[Archive

    Lifescience Database Archive (English)

    Full Text Available NC_002677 gi|15827265 >gi|15827265|ref|NP_301528.1| putative transcriptional regul...ator [Mycobacterium leprae TN] MLTLTIPKQTLPGLPCHADTSDLWFAETPADLECTKTLCANCPIRRPCLEAAMERAEPWGVWGGEIFDRGLIVSRKRPRGRPCNDVVVV

  5. ORF Sequence: NC_002677 [GENIUS II[Archive

    Lifescience Database Archive (English)

    Full Text Available NC_002677 gi|15827349 >gi|15827349|ref|NP_301612.1| hypothetical protein ML0802 [Mycobacterium leprae... TN] MAEMSGVKMAEDVRAEIVASVLEVVVSEGDQIGKGDVLVLLESMKMEIPVLAGVAGIVSKVSVSVGDVIQAGDLIAVIS

  6. ORF Alignment: NC_002677 [GENIUS II[Archive

    Lifescience Database Archive (English)

    Full Text Available ... leprae ... Length = 172 ... Query: 2 ... LELAILGLLIESPMHGYELRKRLTGL...LGAFRAFSYGSLYPALRRMQADRLITGHVAPAGT 61 ... LELAILGLLIESPMHGYELRKRLTGLLGAFRAFSYGSLYPALRRMQADRLITGHVAPAG...T Sbjct: 1 ... LELAILGLLIESPMHGYELRKRLTGLLGAFRAFSYGSLYPALRRMQADRLITGHVAPAGT 60 ... Query: 122 XXXXXXXXXXXXXTRASNS

  7. Tentativas de transmissão da lepra ao homem, por meio de Triatomídeos infectados em doentes lepromatosos: (nota prévia Attempts to transmit Leprosy to man by means of Triatomidae infected in lepromatous cases

    Directory of Open Access Journals (Sweden)

    H. C. de Souza-Araújo

    1944-12-01

    Full Text Available The A.A. started a new series of experiments upon the transmission of Leprosy to man by means of one of the more widespread hematophagi of the hinterland of Brazil, the Triatomidae. Two species of these insects were found naturally infected with Hansen's bacillus in huts of lepers in the interior of the State of Minas Gerais and one of the writers (S.A. upon feeding the same insects on lepromatous cases could obtain two strains of acid-fast bacilli cultures smearing Lowenstein medium with the intestinal contains of the same. The first phase of the experiments lasted five months and the results, partially positive, are here describe. More than one hundred Triatomidae (Triatoma infestans and Panstrongylus megistus bred in the Institute Oswaldo Cruz and fed in normal pigeons until convenient growth were put on lepromatous lesions, which they sucked many times, and them after one or more days they were put to be fed on selected regions of the skin of four negativated cases of leprosy. The arguments in favour and against the possibility of obtaining new lesions of leprosy in such burnet out patients were discussed. The A.A. are not authorized to draw any definite conclusions, but the few facts registered are worth of divulgation, in orther that other workers send their suggestions. Three out of the four volunteers showed moderate local reactions between 1 to 4 days after being sucked by the infected insectes. After five months experiments subcutanous lymph were obtained from the points where the insects have bitten. A very few acid-fast bacilli were found in such material. The patients, being kept in separation from infectious cases, will be followed up during months or a year in order to be detected any suspicious experimental lesions of leprosy.

  8. Lepra y sociedad en la España de la primera mitad del siglo XX: la Colonia Sanatorio de Fontilles (1908-1932) y su proceso de intervención por la Segunda República

    OpenAIRE

    Bernabeu Mestre, Josep; Ballester Artigues, Teresa

    1991-01-01

    La investigación analiza el proceso de intervención de la Leprosería de Fontilles llevado a cabo por la Segunda República en 1932. En una primera parte se exponen las bases institucionales y asistenciales que, caracterizaban a la Colonia Sanatorio de Fontilles en su primera etapa (1908-1932), para estudiar a continuación las razones que condujeron a las autoridades sanitarias del Nuevo Estado a decidir la incautación, y las consecuencias y reacciones científicas, sociales y políticas que aque...

  9. Confiabilidad y concordancia de dos escalas de lectura de baciloscopias para clasificación y seguimiento del tratamiento con múltiples medicamentos de los pacientes con lepra

    Directory of Open Access Journals (Sweden)

    Claudia Lucía Colorado

    2011-03-01

    Conclusiones. La concordancia entre observadores obtenida para ambas escalas fue excelente, al igual que la correlación de la concordancia de los índices bacilares calculados con los dos métodos. Con los puntos de corte establecidos, se obtuvo un nivel de concordancia excelente, lo que garantiza que las escalas son intercambiables a la hora de establecer si la carga bacilar es alta o baja.

  10. Protein splicing and its evolution in eukaryotes

    Directory of Open Access Journals (Sweden)

    Starokadomskyy P. L.

    2010-02-01

    Full Text Available Inteins, or protein introns, are parts of protein sequences that are post-translationally excised, their flanking regions (exteins being spliced together. This process was called protein splicing. Originally inteins were found in prokaryotic or unicellular eukaryotic organisms. But the general principles of post-translation protein rearrangement are evolving yielding different post-translation modification of proteins in multicellular organisms. For clarity, these non-intein mediated events call either protein rearrangements or protein editing. The most intriguing example of protein editing is proteasome-mediated splicing of antigens in vertebrates that may play important role in antigen presentation. Other examples of protein rearrangements are maturation of Hg-proteins (critical receptors in embryogenesis as well as maturation of several metabolic enzymes. Despite a lack of experimental data we try to analyze some intriguing examples of protein splicing evolution.

  11. Split and Splice Approach for Highly Selective Targeting of Human NSCLC Tumors

    Science.gov (United States)

    2014-10-01

    for Stx2A-specific intracellular delivery mechanism by replacing four lysine residues in its C-terminal part with arginines and confirming that the...Fig. 3. Substitution of all 4 lysines in the C-terminal intein part by arginines preserves its trans-splicing activity in vivo as revealed by using...not directly applicable for in vivo trans-splicing of Stx2A split toxin due to the presence of four lysines in the C-terminal part of the intein

  12. 12mer Phage Display Peptide Library

    African Journals Online (AJOL)

    efficacy in the production of anti-M. leprae antibodies in an animal model. Methods: Blood samples were ... and western blot. anti-leprae antibodies in various dilutions and were found to be serological active. Sequencing of the isolated peptides .... Serial dilutions of phage were prepared in LB broth (1 % Yeast extract, ...

  13. From genome-based in silico predictions to ex vivo verification of leprosy diagnosis

    NARCIS (Netherlands)

    Geluk, Annemieke; Spencer, John S.; Bobosha, Kidist; Pessolani, Maria C. V.; Pereira, Geraldo M. B.; Banu, Sayera; Honoré, Nadine; Reece, Stephen T.; MacDonald, Murdo; Sapkota, Bishwa Raj; Ranjit, Chaman; Franken, Kees L. M. C.; Zewdie, Martha; Aseffa, Abraham; Hussain, Rabia; Stefani, Mariane M.; Cho, Sang-Nae; Oskam, Linda; Brennan, Patrick J.; Dockrell, Hazel M.

    2009-01-01

    The detection of hundreds of thousands of new cases of leprosy every year suggests that transmission of Mycobacterium leprae infection still continues. Unfortunately, tools for identification of asymptomatic disease and/or early-stage M. leprae infection (likely sources of transmission) are lacking.

  14. PATTERNS OF SEVEN AND COMPLICATED MALARIA IN CHILDREN

    African Journals Online (AJOL)

    GB

    2015-10-04

    Oct 4, 2015 ... Mycobacterium leprae. The causative agent affects peripheral nerves and causes damage by binding with Schwann cells which are important for conducting nerve impulses (1, 2). The interaction of M. leprae with Schwann cells causes irreversible loss of peripheral nerve tissue followed by disabilities (3).

  15. Overview of the recombinant proteins purification by affinity tags and ...

    African Journals Online (AJOL)

    From protein within isolation process which the same matter increases labor costs further and prevents application of these tags in industrial scale. Therefore proper replacement is emphasized for enzymatic removal of purification tags. Keywords: protein purification; recombinant proteins; self-cleavable tags; Intein tags; ...

  16. Extending the scope of site-specific cysteine bioconjugation by appending a prelabeled cysteine tag to proteins using protein trans-splicing.

    Science.gov (United States)

    Dhar, Tulika; Kurpiers, Thomas; Mootz, Henning D

    2011-01-01

    Incorporating synthetic probes site-specifically into proteins is of central interest in several areas of biotechnology and protein chemistry. Bioconjugation techniques provide a simple and effective means of chemically modifying a protein. In particular, covalent chemical modifications of cysteine residues belong to one of the most important reactions due to the unique reactivity of its thiol moiety and the relatively low abundance of this amino acid in proteins. However, such types of modifications cannot be performed in a regioselective fashion when one or more additional cysteines are present. To address this limitation, we have developed an approach where a short cysteine-containing tag (Cys-Tag) fused to one part of a split intein and modified at its sulfhydryl group can be used to label proteins by trans-splicing with a protein of interest (POI) fused to the other half of the split intein. In this way, it is possible to selectively label a protein containing multiple cysteines. The artificially split Mycobacterium xenopi GyrA intein and the Synechocystis sp. DnaB intein were highly suitable for this purpose and were successfully used for the labeling of several proteins. This approach enables a simple route for labeling proteins by site-specific cysteine bioconjugation with any one of several commercially available cysteine-modifying probes.

  17. Production of Nα-acetylated thymosin α1 in Escherichia coli

    Directory of Open Access Journals (Sweden)

    Fang Hongqing

    2011-04-01

    Full Text Available Abstract Background Thymosin α1 (Tα1, a 28-amino acid Nα-acetylated peptide, has a powerful general immunostimulating activity. Although biosynthesis is an attractive means of large-scale manufacture, to date, Tα1 can only be chemosynthesized because of two obstacles to its biosynthesis: the difficulties in expressing small peptides and obtaining Nα-acetylation. In this study, we describe a novel production process for Nα-acetylated Tα1 in Escherichia coli. Results To obtain recombinant Nα-acetylated Tα1 efficiently, a fusion protein, Tα1-Intein, was constructed, in which Tα1 was fused to the N-terminus of the smallest mini-intein, Spl DnaX (136 amino acids long, from Spirulina platensis, and a His tag was added at the C-terminus. Because Tα1 was placed at the N-terminus of the Tα1-Intein fusion protein, Tα1 could be fully acetylated when the Tα1-Intein fusion protein was co-expressed with RimJ (a known prokaryotic Nα-acetyltransferase in Escherichia coli. After purification by Ni-Sepharose affinity chromatography, the Tα1-Intein fusion protein was induced by the thiols β-mercaptoethanol or d,l-dithiothreitol, or by increasing the temperature, to release Tα1 through intein-mediated N-terminal cleavage. Under the optimal conditions, more than 90% of the Tα1-Intein fusion protein was thiolyzed, and 24.5 mg Tα1 was obtained from 1 L of culture media. The purity was 98% after a series of chromatographic purification steps. The molecular weight of recombinant Tα1 was determined to be 3107.44 Da by mass spectrometry, which was nearly identical to that of the synthetic version (3107.42 Da. The whole sequence of recombinant Tα1 was identified by tandem mass spectrometry and its N-terminal serine residue was shown to be acetylated. Conclusions The present data demonstrate that Nα-acetylated Tα1 can be efficiently produced in recombinant E. coli. This bioprocess could be used as an alternative to chemosynthesis for the production

  18. Dicty_cDB: Contig-U01389-1 [Dicty_cDB

    Lifescience Database Archive (English)

    Full Text Available :none) Sinorhizobium sp. CCNWGS114 recomb... 176 1e-42 AM182138_1( AM182138 |pid:none) Sinorhizobium saheli ...partial recA ... 176 1e-42 AJ294380_1( AJ294380 |pid:none) Sinorhizobium saheli partial recA ... 176 1e-42 B...7 1e-42 (Q9XBC1) RecName: Full=Protein recA; AltName: Full=Recombinase A; 176 1e-42 AM182140_1( AM182140 |pid:none) Sinorhizobium sah...eli partial recA ... 176 1e-42 FJ619312_1( FJ619312 |pid

  19. Abordaje logoterapéutico para la prevención de recaídas en el consumo de drogas, en los residentes de la Comunidad Terapéutica Aprender a Vivir, en el periodo de marzo a julio del 2017

    OpenAIRE

    Bossano Muñoz, Ronny Martín; Onofa Villarreal, Abigail Elizabeth

    2018-01-01

    The following work represents the systematization of a model of logotherapeutic intervention applied in the “Comunidad Terapeutica Aprender A Vivir”, in the period from March to July 2017. The need to carry out this project arises with the intention of complementing the work done in this institution. from a new humanistic focus, promoting a person-centered vision and this way enabling an alternative strategy for the prevention of relapses in drug use. The model that allows t...

  20. Validation of qPCR Methods for the Detection of Mycobacterium in New World Animal Reservoirs

    Science.gov (United States)

    Housman, Genevieve; Malukiewicz, Joanna; Boere, Vanner; Grativol, Adriana D.; Pereira, Luiz Cezar M.; Silva, Ita de Oliveira e; Ruiz-Miranda, Carlos R.; Truman, Richard; Stone, Anne C.

    2015-01-01

    Zoonotic pathogens that cause leprosy (Mycobacterium leprae) and tuberculosis (Mycobacterium tuberculosis complex, MTBC) continue to impact modern human populations. Therefore, methods able to survey mycobacterial infection in potential animal hosts are necessary for proper evaluation of human exposure threats. Here we tested for mycobacterial-specific single- and multi-copy loci using qPCR. In a trial study in which armadillos were artificially infected with M. leprae, these techniques were specific and sensitive to pathogen detection, while more traditional ELISAs were only specific. These assays were then employed in a case study to detect M. leprae as well as MTBC in wild marmosets. All marmosets were negative for M. leprae DNA, but 14 were positive for the mycobacterial rpoB gene assay. Targeted capture and sequencing of rpoB and other MTBC genes validated the presence of mycobacterial DNA in these samples and revealed that qPCR is useful for identifying mycobacterial-infected animal hosts. PMID:26571269

  1. Validation of qPCR Methods for the Detection of Mycobacterium in New World Animal Reservoirs.

    Science.gov (United States)

    Housman, Genevieve; Malukiewicz, Joanna; Boere, Vanner; Grativol, Adriana D; Pereira, Luiz Cezar M; Silva, Ita de Oliveira; Ruiz-Miranda, Carlos R; Truman, Richard; Stone, Anne C

    2015-11-01

    Zoonotic pathogens that cause leprosy (Mycobacterium leprae) and tuberculosis (Mycobacterium tuberculosis complex, MTBC) continue to impact modern human populations. Therefore, methods able to survey mycobacterial infection in potential animal hosts are necessary for proper evaluation of human exposure threats. Here we tested for mycobacterial-specific single- and multi-copy loci using qPCR. In a trial study in which armadillos were artificially infected with M. leprae, these techniques were specific and sensitive to pathogen detection, while more traditional ELISAs were only specific. These assays were then employed in a case study to detect M. leprae as well as MTBC in wild marmosets. All marmosets were negative for M. leprae DNA, but 14 were positive for the mycobacterial rpoB gene assay. Targeted capture and sequencing of rpoB and other MTBC genes validated the presence of mycobacterial DNA in these samples and revealed that qPCR is useful for identifying mycobacterial-infected animal hosts.

  2. Leprosy Specific Orofacial Aspects

    Directory of Open Access Journals (Sweden)

    Vathsala Naik

    2011-01-01

    Full Text Available Leprosy is a chronic infection caused by Mycobactenum leprae, GHA- Hansen first identified the organism in 1873, so called Hansen disease. Mycobacterium leprae is a bacillus that presents a peculiar tropism for the skin and peripheral nerves. The upper airway has a great importance as a route of M. Leprae infection. The clinical spectrum of leprosy ranges from the tuberculoid form (TT to the disseminative and progressive lepromatous form (LL. Cell-mediated immunity is considered to be the crucial defence against the disease and the magnitude of this immunity defines the extent of the disease- Facial lesions in leprosy can occur in all form of the disease and also in lepra reaction, oral lesions are rare but, when present, occur in the lepromatous form

  3. Síntesis de los problemas sanitarios en colombia

    OpenAIRE

    Muñoz, Laurentino

    2010-01-01

    Panorama de la salud; Trabajo; Parasitosis intestinal; Paludismo; Tuberculosis; Enfermedades venéreas; Lepra -- Comparación; Mortalidad infantil; Condiciones antihigiénicas de la habitación; Conciencia sanitaria

  4. analysis, diagnosis and prognosis of leprosy utilizing fuzzy classifier ...

    African Journals Online (AJOL)

    TRIPPLEJO2K2

    leprae to humans: chimpanzees, mangabey monkeys, and nine-banded armadillos. The early signs and symptoms of leprosy are very subtle and occur slowly (usually over years), which includes; numbness (loss of temperature sensation), skin ...

  5. Recognition of DNA damage and induction of repair proteins in deinococcus radiodurans

    Energy Technology Data Exchange (ETDEWEB)

    Satoh, Katsuya; Kikuchi, Masahiro; Narumi, Issay [Japan Atomic Energy Research Inst., Takasaki, Gunma (Japan). Takasaki Radiation Chemistry Research Establishment

    2002-03-01

    In Escherichia coli, the inducible DNA repair system (the SOS system) is regulated by two key proteins; RecA and LexA. Although SOS-like processes have been documented in a wide variety of eubacterial species. The involvement of RecA and LexA in the SOS response is poorly understood in Deinococcus radiodurans. As expression of the deinococcal recA is enhanced after irradiation, the recA seems to be a member of a DNA damage response regulon in D. radiodurans. By investigating the function of deinococcal proteins, we found that LexA is not involved in the regulation of RecA in D. radiodurans. This led us to speculate that D. radiodurans has an alternative DNA damage response mechanism with which to control recA expression. We discovered that a novel protein regulates the expression of recA. The novel regulatory protein (designated PprI) also controls the induction of pprA following irradiation. We also found that the co-protease activity rather than recombination activity of RecA contributes to the radiation resistance in D. radiodurans. The D. radiodurans genome encodes a second but diverged copy of LexA (designated LexA2). Analysis of LexA2 indicated that still-unknown repair gene are down-regulated by LexA2. Thus, D. radiodurans possesses unique mechanisms of DNA damage recognition and repair gene induction. (author)

  6. Bacillus halodurans RecA-DNA binding and RecAmediated ...

    African Journals Online (AJOL)

    Abstract. In Escherichia coli, RecA protein catalyzes DNA pairing and strand exchange activities essential for genetic recombination. This is critical for normal cellular function under conditions that lead to altered. DNA metabolism and DNA damage. The RecA proteins of E. coli and Bacillus halodurans both can bind to DNA ...

  7. RecA-mediated cleavage reaction of Lambda repressor and DNA ...

    African Journals Online (AJOL)

    PRECIOUS

    2010-01-11

    Jan 11, 2010 ... 00237 77 05 25 11. Accepted 4 January, 2010. DNA pairing and strand exchange activities are essential for genetic recombination. When DNA is damaged, RecA ... and the ability of RecA to promote DNA strand exchange. It was observed ..... hydrolyze the half of ATP after 6 min (t1) and 12 min (t2) after.

  8. Validation of qPCR Methods for the Detection of Mycobacterium in New World Animal Reservoirs

    OpenAIRE

    Housman, Genevieve; Malukiewicz, Joanna; Boere, Vanner; Grativol, Adriana D.; Pereira, Luiz Cezar M.; Silva, Ita de Oliveira e; Ruiz-Miranda, Carlos R.; Truman, Richard; Stone, Anne C.

    2015-01-01

    Zoonotic pathogens that cause leprosy (Mycobacterium leprae) and tuberculosis (Mycobacterium tuberculosis complex, MTBC) continue to impact modern human populations. Therefore, methods able to survey mycobacterial infection in potential animal hosts are necessary for proper evaluation of human exposure threats. Here we tested for mycobacterial-specific single- and multi-copy loci using qPCR. In a trial study in which armadillos were artificially infected with M. leprae, these techniques were ...

  9. Protection of DNA damage by radiation exposure

    International Nuclear Information System (INIS)

    Lee, Jeong Ho; Kim, In Gyu; Lee, Kang Suk; Kim, Kug Chan; Oh, Tae Jung

    1998-12-01

    The SOS response of Escherichia coli is positively regulated by RecA. To examine the effects of polyamines on The SOS response of E. Coli, we investigated the expression of recA gene in polyamine-deficient mutant and wild type carrying recA'::lacZ fusion gene. As a result, recA expression by mitomycin C is higher in wild type than that of polyamine-deficient mutant, but recA expression by UV radiation is higher in wild type than of mutant. We also found that exogenous polyamines restored the recA expression in the polyamine-deficient mutant to the wild type level. These results proposed that polyamines play an important role in mechanism of intracellular DNA protection by DNA damaging agents

  10. Protection of DNA damage by radiation exposure

    Energy Technology Data Exchange (ETDEWEB)

    Lee, Jeong Ho; Kim, In Gyu; Lee, Kang Suk; Kim, Kug Chan; Oh, Tae Jung

    1998-12-01

    The SOS response of Escherichia coli is positively regulated by RecA. To examine the effects of polyamines on The SOS response of E. Coli, we investigated the expression of recA gene in polyamine-deficient mutant and wild type carrying recA'::lacZ fusion gene. As a result, recA expression by mitomycin C is higher in wild type than that of polyamine-deficient mutant, but recA expression by UV radiation is higher in wild type than of mutant. We also found that exogenous polyamines restored the recA expression in the polyamine-deficient mutant to the wild type level. These results proposed that polyamines play an important role in mechanism of intracellular DNA protection by DNA damaging agents.

  11. TER HANSENIASE: PERCEPÇÕES DE PESSOAS EM TRATAMENTO

    Directory of Open Access Journals (Sweden)

    JOYCE MAZZA NUNES

    2008-01-01

    Full Text Available La lepra es una enfermedad milenaria, caracterizada como un problema de salud pública. Desarrollamos este estudio con el objetivo de conocer la percepción sobre su enfermedad de las personas en tratamiento para lepra. Se trata de un estudio con planteo cualitativo, desarrollado a través de un grupo focal en el mes de agosto de 2005 con ocho personas en tratamiento para lepra multibacilar en la Unidad Básica de Salud en Sobral-CE. Las declaraciones señalaron que la lepra es vista como una enfermedad mala, que trae tristeza, miedo, vergüenza y discriminación. Ser portador de lepra ocasionó sufrimiento psíquico y sentimientos de impotencia, hombres y mujeres tuvieron sus roles sociales amenazados. Sugerimos que se idealicen alternativas que atiendan no sólo la eliminación de la lepra como un problema de salud pública, sino también, como un rescate de la ciudadanía y del respeto por esas personas.

  12. Expression of Recombinant Proteins with Uniform N-Termini

    OpenAIRE

    Király, Orsolya; Guan, Lan; Sahin-Tóth, Miklós

    2011-01-01

    Heterologously expressed proteins in Escherichia coli may undergo unwanted N-terminal processing by methionine and proline aminopeptidases. To overcome this problem, we present a system where the gene of interest is cloned as a fusion to a self-splicing mini-intein. Furthermore, this fusion construct is expressed in an engineered Escherichia coli strain from which the pepP gene coding for aminopeptidase P has been deleted. We describe a protocol using human cationic trypsinogen as an example ...

  13. Ensaios sôbre lepra experimental: inoculações de três amostras de bacilos ácido-álcool resistentes (amostras "Chaves II", "Emilia" e "Hecke" isoladas de leprosos, em trinta doentes da Colônia Mirueira

    Directory of Open Access Journals (Sweden)

    H. C. de Souza-Araujo

    1951-03-01

    Full Text Available Com o fim de produzir o fenômeno de KOCH, os A. A. inocularam em 30 leprosos da Colônia Mirueira (Recife, de várias idades e formas clínicas, emulsões vivas de três culturas de bacilos ácido-álcool resistentes isolados de leprosos pro um dêles (S.A.. As doses inoculadas foram de 0,2 cc., por via intradérmica, em cada doente, das amostras "CII", "E" e "H" e mais da Leprolina S.A. (antígeno morto. No 10º dia da inoculação verificou-se que 24 dos 30 pacientes tiveram reação geral intensa; 2, reação moderada e 4, nenhuma reação geral. 16 dos 30 tiveram reação leprótica, sendo 10 em casos ativos (lepromatosos e 6 em inativos, e 17 dos 30 tiveram adenopatias inguinais. O inóculo "CII" produziu escaras de 1 x 1 e 2 x 2 cm. de diâmetro, com destruição total da pele, nos 30 pacientes (o total dêles; o inóculo "E" produziu escaras de igual intensidade em 29, o inóculo "H", escaras muitos mais benignas em 23, e a Leprolina em 10, naturalmente por ação concomitante de um dos outros três inóculos. No 10º dia foram semeadas em meio de LOEWENSTEIN secreções das escaras de sete dos 30 doentes, num total de 20 tubos, dos quais 19 produziram retroculuras, a amioria contaminada por fungos ou por bactérias cianófilas. De um doente foi obtido retrocultura cromogênica da escara produzida na intradérmoreação pela "Leprolina S. A.", macro e microscòpricamente indiferencável das amostras "CII" e "E". Aliás, pela extensiva experimentação feita com estas duas amostras, estamos nos inclinando por considerá-las como idênticas. No 18º dia da inoculação foram feitos 30 esfregaços de secreções de lesões experimentais de 13 doentes, com 15 resultados positivos (50%, apesar do exame tardio. As morfologias macro e microscópica das retroculturas obtidas em Recife confirmam os caracteres descritos nas culturas originais. Dêste rápido ensaio se conclui que a maioria dos pacientes apresentou o fenômeno de KOCH parcial ou integral, com as clássicas reações gerais, focais e locais. A falta de recursos de laboratório na Colônia não permitiu melhor aproveitamento de tão precioso material experimental, e por isso êste trabalho apresenta várias lacunas.With the aim to produce the KOCH's phenomenon, the A. A. incoulated 30 lepers of the Colonia Mirueira (Recife, from various ages and clinical types, with alive suspensios of three strains of acid-fast bacilli isolated from lepers. Each volunteer received three intradermical inoculations in his thighs, of 0.2 c.c. each from strains "Chaves II", "Emília", and "Hecke" and in his left forea rom 0.2 c.c. of Leprolina S.A. (killed antigen. At the 10th day of inoculation were verified that 24 out of the 30 volunteers had severe general reaction, 2, moderate ones and 4, nothing. 16 out of the 30 had leprosy reaction, being 10 lepromatous cases and 6 inactive oens. The inoculum "C II" produced craters, of 1 x 1 and 2 x 2 cm. diameter in the 30 patients (total, craters with complete destruction of the skin; the inoculum "E" did the same in 29, of equal severety and the inoculum "H" did only in 23, but lesions of minor severety and the "L" produced craters in ten patients, evidently due to the concomitant action of one of the three inocula. At the same 10th day were inoculated secretions onto 20 tubes of LOEWENSTEIN medium of seven patients. 18 days later were seen developpment of retrocultures in 19 ou 20 tubes, the mojority being contaminated by fungs or cyanophile bacteria. From one patient (nº 16 was recovered a chromogenic culture from his Leprolin-test, macro and microscopically undistinguishable from strain "C II" or "E", which, based on a large experimentation the senior writer is inclined to consider as identical. At the 18 th day were made smears from lesions of 13 patients, 30 in total with 15 positive (50% for one or another of the inocula. The macro and microscopical morphologies of the recovered cultures (retrocultrues coincide with the characteristics of the original cultures from which the inocula were made. In conclusion, the majority of the patients showed partial or integral KOCH's prenomenon, with its classical general, local and focal reactions. By lack of laboratory resource, at the leper Colony, the precious material available from this series of experiments were not used, resulting in omissions of various kind in this paper.

  14. Poderá o carrapato transmitir a lepra?: mais quatro amostras de culturas de bacilos acido-alcool resistentes obtidas de carrapatos (2 de "Amblyomma cajennense"e 2 de "Boophilus microplus" infectados em leprosos do Paraná: 3ª nota May leprosy be transmitted by ticks?: third note

    Directory of Open Access Journals (Sweden)

    H. C. de Souza-Araujo

    1942-01-01

    Full Text Available The AA. carried out experiments in the leprosarium São Roque, State of Paraná, South Brazil, to verify if the cattle tick Boophilus microplus could be experimentally infected in lepers, which was true. The AA. Tried also to be ascertained if Boophilus microplus and Amblyomma cajennense could change of hosts during their feedings which was true, both ticks continue feeding, the last species for many days, after being transferred from one to another leper. The junior A. describes in full their experiments and also a dermatites caused by tick bites. The senior A. brought to Rio de Janeiro most of the infected ticks for examination, which revealed a very high positivity. He smeared the sediments of lots of both species of ticks in Loewenstein medium and after a variable periode of incubation at 37° C. he obtained four new samples of cultures of acid-fast organisms, two from Amblyomma cajennense and two from Boophilus microplus. These cultures are being studied and will be inoculated into laboratory animals. The senior A. inoculated new batches of white rats with sediments of many ticks infected in lepers. Various hypotheses of both previous notes upon the subject now are verified facts. The A. is accumulating facts to draw the conclusions in the future. He also suggested the leprosy workers in the interior of the country to cooperate with him in such important studies, specially in the habitat of lepers in the rural zones of various States.

  15. Consulta de enfermagem ao portador de Hanseníase: proposta de um instrumento para aplicação do processo de enfermagem Consulta de enfermería al portador de la Lepra: propuesta de una herramienta para aplicación del proceso de enfermería Nursing consultation for Leprosy patients: proposal of an instrument for nursing process application

    OpenAIRE

    Marli Teresinha Cassamassimo Duarte; Jairo Aparecido Ayres; Janete Passuto Simonetti

    2008-01-01

    Objetivou-se relatar a experiência da consulta de enfermagem junto aos portadores de hanseníase, realizada em unidade de atenção primária à saúde de uma Universidade do interior do Estado de São Paulo, bem como apresentar o instrumento utilizado. Essa atividade é realizada por dois enfermeiros, que atuam por mais de duas décadas no programa, acumulando, assim, experiência no cuidado a esses indivíduos. Avaliam-se a eficiência dos instrumentos quanto à forma e conteúdo, possibilitando o levant...

  16. Effects of virus and host genes on recombination among ultraviolet-irradiated bacteriophage T4

    International Nuclear Information System (INIS)

    Priemer, M.M.; Chan, V.L.

    1978-01-01

    The influence of the polA, uvrA, and recA genes of Escherichia coli on recombination among ultraviolet-irradiated T4 bacteriophages was determined with respect to recombination between rII markers and phage yield. The polA and uvrA gene products have no effect on these two aspects of phage DNA metabolism. A recA mutation does not significantly alter rII recombination frequencies in irradiated phage crosses, nor does it greatly change the yield of infectious particles in wild-type phage crosses or crosses in which the phage strains possess the v mutation. However, the same cross experiment performed with a pair of T4x mutants in a recA host demonstrates an 84% reduction in the phage yield in an unirradiated control cross. Furthermore, with increasing doses of uv irradiation, phage productivity of the T4x mutant declines at an accelerated rate compared to T4x + strains crossed in recA cells. Multiplicity reactivation experiments in which wild-type or recombination-deficient (x or y) T4 phages infect wild-type or recombination-deficient (recA) host cells show that irradiated phages can only be reactivated in recA + hosts, regardless of the bacteriophage genotype. These results indicate the involvement of the E. coli recA gene product in normal T4 replication and multiplicity reactivation

  17. Engineering a recyclable elastin-like polypeptide capturing scaffold for non-chromatographic protein purification.

    Science.gov (United States)

    Liu, Fang; Chen, Wilfred

    2013-01-01

    Previously, we reported a non-chromatographic protein purification method exploiting the highly specific interaction between the dockerin and cohesin domains from Clostridium thermocellum and the reversible aggregation property of elastin-like polypeptide (ELP) to provide fast and cost-effective protein purification. However, the bound dockerin-intein tag cannot be completely dissociated from the ELP-cohesin capturing scaffold due to the high binding affinity, resulting in a single-use approach. In order to further reduce the purification cost by recycling the ELP capturing scaffold, a truncated dockerin domain with the calcium-coordinating function partially impaired was employed. We demonstrated that the truncated dockerin domain was sufficient to function as an effective affinity tag, and the target protein was purified directly from cell extracts in a single binding step followed by intein cleavage. The efficient EDTA-mediated dissociation of the bound dockerin-intein tag from the ELP-cohesin capturing scaffold was realized, and the regenerated ELP capturing scaffold was reused in another purification cycle without any decrease in the purification efficiency. This recyclable non-chromatographic based affinity method provides an attractive approach for efficient and cost-effective protein purification. © 2013 American Institute of Chemical Engineers.

  18. RNA-Mediated cis Regulation in Acinetobacter baumannii Modulates Stress-Induced Phenotypic Variation.

    Science.gov (United States)

    Ching, Carly; Gozzi, Kevin; Heinemann, Björn; Chai, Yunrong; Godoy, Veronica G

    2017-06-01

    In the nosocomial opportunistic pathogen Acinetobacter baumannii , RecA-dependent mutagenesis, which causes antibiotic resistance acquisition, is linked to the DNA damage response (DDR). Notably, unlike the Escherichia coli paradigm, recA and DDR gene expression in A. baumannii is bimodal. Namely, there is phenotypic variation upon DNA damage, which may provide a bet-hedging strategy for survival. Thus, understanding recA gene regulation is key to elucidate the yet unknown DDR regulation in A. baumannii Here, we identify a structured 5' untranslated region (UTR) in the recA transcript which serves as a cis -regulatory element. We show that a predicted stem-loop structure in this 5' UTR affects mRNA half-life and underlies bimodal gene expression and thus phenotypic variation in response to ciprofloxacin treatment. We furthermore show that the stem-loop structure of the recA 5' UTR influences intracellular RecA protein levels and, in vivo , impairing the formation of the stem-loop structure of the recA 5' UTR lowers cell survival of UV treatment and decreases rifampin resistance acquisition from DNA damage-induced mutagenesis. We hypothesize that the 5' UTR allows for stable recA transcripts during stress, including antibiotic treatment, enabling cells to maintain suitable RecA levels for survival. This innovative strategy to regulate the DDR in A. baumannii may contribute to its success as a pathogen. IMPORTANCE Acinetobacter baumannii is an opportunistic pathogen quickly gaining antibiotic resistances. Mutagenesis and antibiotic resistance acquisition are linked to the DNA damage response (DDR). However, how the DDR is regulated in A. baumannii remains unknown, since unlike most bacteria, A. baumannii does not follow the regulation of the Escherichia coli paradigm. In this study, we have started to uncover the mechanisms regulating the novel A. baumannii DDR. We have found that a cis -acting 5' UTR regulates recA transcript stability, RecA protein levels, and DNA

  19. Prophage λ induction by ionizing radiation of different LETs

    International Nuclear Information System (INIS)

    Bonev, M.N.; Kozubek, S.; Krasavin, E.A.; Amirtaev, K.G.

    1988-01-01

    The λ prophage induction caused by γ-irradiation and accelerated heavy ions with different LET was studied in variety Escherichia coli strains. The induction frequency on the dose I(D) shaped a curve with a maximum in the strains which possess recA + /lexA + genotype. The inductivity of these strains increases as well as LET and an alteration poor → rich media does it. Unlike I(D) for recA + /lexA + , the dependence I(D) for recA, lexA and recBC strains was a constant. 15 refs.; 6 figs.; 3 tabs

  20. Formation of active inclusion bodies induced by hydrophobic self-assembling peptide GFIL8.

    Science.gov (United States)

    Wang, Xu; Zhou, Bihong; Hu, Weike; Zhao, Qing; Lin, Zhanglin

    2015-06-16

    In the last few decades, several groups have observed that proteins expressed as inclusion bodies (IBs) in bacteria could still be biologically active when terminally fused to an appropriate aggregation-prone partner such as pyruvate oxidase from Paenibacillus polymyxa (PoxB). More recently, we have demonstrated that three amphipathic self-assembling peptides, an alpha helical peptide 18A, a beta-strand peptide ELK16, and a surfactant-like peptide L6KD, have properties that induce target proteins into active IBs. We have developed an efficient protein expression and purification approach for these active IBs by introducing a self-cleavable intein molecule. In this study, the self-assembling peptide GFIL8 (GFILGFIL) with only hydrophobic residues was analyzed, and this peptide effectively induced the formation of cytoplasmic IBs in Escherichia coli when terminally attached to lipase A and amadoriase II. The protein aggregates in cells were confirmed by transmission electron microscopy analysis and retained ~50% of their specific activities relative to the native counterparts. We constructed an expression and separation coupled tag (ESCT) by incorporating an intein molecule, the Mxe GyrA intein. Soluble target proteins were successfully released from active IBs upon cleavage of the intein between the GFIL8 tag and the target protein, which was mediated by dithiothreitol. A variant of GFIL8, GFIL16 (GFILGFILGFILGFIL), improved the ESCT scheme by efficiently eliminating interference from the soluble intein-GFIL8 molecule. The yields of target proteins at the laboratory scale were 3.0-7.5 μg/mg wet cell pellet, which is comparable to the yields from similar ESCT constructs using 18A, ELK16, or the elastin-like peptide tag scheme. The all-hydrophobic self-assembling peptide GFIL8 induced the formation of active IBs in E. coli when terminally attached to target proteins. GFIL8 and its variant GFIL16 can act as a "pull-down" tag to produce purified soluble proteins with

  1. Radiation damage and repair in cells and cell components. Progress report: third new contract year

    International Nuclear Information System (INIS)

    Fluke, D.J.; Pollard, E.C.

    1980-01-01

    Research progress for 1979-1980 is reported. Projects discussed include the process of radiation-induced repair, Weigle-reactivation, induced radioresistance, the induction of the recA gene product, uv mutagenesis, and the induction of lambda

  2. Simple method for identification of plasmid-coded proteins

    International Nuclear Information System (INIS)

    Sancar, A.; Hack, A.M.; Rupp, W.D.

    1979-01-01

    Proteins encoded by plasmid DNA are specifically labeled in uv-irradiated cells of Escherichia coli carrying recA and uvrA mutations because extensive degradation of the chromosome DNA occurs concurrently with amplification of plasmid DNA

  3. Radiation damage and repair in cells and cell components. Progress report: third new contract year

    Energy Technology Data Exchange (ETDEWEB)

    Fluke, D.J.; Pollard, E.C.

    1980-01-01

    Research progress for 1979-1980 is reported. Projects discussed include the process of radiation-induced repair, Weigle-reactivation, induced radioresistance, the induction of the recA gene product, uv mutagenesis, and the induction of lambda. (ACR)

  4. The role of ERBB2 gene polymorphisms in leprosy susceptibility

    Directory of Open Access Journals (Sweden)

    Jamile Leão Rêgo

    2015-03-01

    Full Text Available Mycobacterium leprae infects skin and peripheral nerves causing deformities and disability. The M. leprae bacterium binds to ErbB2 on the Schwann cell surface causing demyelination and favoring spread of the bacilli and causing nerve injury. Polymorphisms at the ERBB2 gene were previously investigated as genetic risk factors for leprosy in two Brazilian populations but with inconsistent results. Herein we extend the analysis of ERBB2 variants to a third geographically distinct population in Brazil. Our results show that there is no association between the genotyped SNPs and the disease (p > 0.05 in this population. A gene set or pathway analysis under the genomic region of ERBB2 will be necessary to clarify its regulation under M. leprae stimulus.

  5. Avaliação da reação de mitsuda em pacientes virchovianos inativos antes e após imunoterapia

    Directory of Open Access Journals (Sweden)

    Maria Sueli Parreira de Arruda

    1995-09-01

    Full Text Available Neste estudo investigou-se o potencial imunomodulador do levamisole e da mistura BCG/Mycobacterium leprae em pacientes virchovianos inativos, utilizando como parâmetro a reação de Mitsuda. Vinte pacientes, classificados como Mitsuda histologicamente negativos há 10 anos, foram divididos em três grupos: cinco pacientes que foram somente reavaliados frente a mitsudina: oito pacientes que receberam levamisole e, sete que receberam a mistura de BCG vivo mais M. leprae morto. Os resultados mostraram que: 1 o levamisole não alterou a reatividade à mitsudina em nenhum dos casos estudados; 2 as modificações da reatividade verificadas com o uso da mistura (tres casos ou aquelas que ocorreram espontaneamente (tres casos foram sempre de pequena amplitude e refletiram variações próprias de pacientes com algum grau de resistência ao Mycobacterium leprae.

  6. Inhibition of the SOS response of Escherichia coli by the Ada protein

    International Nuclear Information System (INIS)

    Vericat, J.A.; Guerrero, R.; Barbe, J.

    1988-01-01

    Induction of the adaptive response by N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) caused a decrease in the UV-mediated expression of both recA and sfiA genes but not of the umuDC gene. On the other hand, the adaptive response did not affect the temperature-promoted induction of SOS response in a RecA441 mutant. The inhibitory effect on the UV-triggered expression of the recA and sfiA genes was not dependent on either the alkA gene or the basal level of RecA protein, but rather required the ada gene. Furthermore, an increase in the level of the Ada protein, caused by the runaway plasmid pYN3059 in which the ada gene is regulated by the lac promoter, inhibited UV-mediated recA gene expression even in cells to which the MNNG-adaptive treatment had not been applied. This inhibitory effect of the adaptive pretreatment was not observed either in RecBC- strains or in RecBC mutants lacking exonuclease V-related nuclease activity. However, RecF- mutants showed an adaptive response-mediated decrease in UV-promoted induction of the recA gene

  7. Rhizobium anhuiense as the predominant microsymbionts of Lathyrus maritimus along the Shandong Peninsula seashore line.

    Science.gov (United States)

    Li, Yan; Wang, En Tao; Liu, Yajing; Li, Xiangyue; Yu, Bing; Ren, Chenggang; Liu, Wei; Li, Yunzhao; Xie, Zhihong

    2016-09-01

    Beach pea [Lathyrus maritimus Bigelow, or Lathyrus japonicus subsp. maritimus (L.) P.W. Ball] is a wild legume distributed on the seashore line, and the rhizobia nodulating with this plant have been reported only rarely. In order to reveal the diversity of beach pea rhizobia on the seashore line of Shandong Peninsula, China, a total of 124 bacterial strains were isolated from the root nodules of beach pea plants collected from five sites. All the isolates were divided into five recA types after screening by recA gene sequence analysis and they consisted of Rhizobium anhuiense covering 122 symbiotic isolates in three recA types, as well as two single isolates Rhizobium sp. and Rhizobium lusitanum representing distinct recA types. The recA genotype III of R. anhuiense (103 isolates) represented by strain YIC11270 was dominant at all five sampling sites. Identical symbiotic genes (nodC and nifH) were detected in the three recA genotypes of R. anhuiense isolates that were closely related to those of the pea and faba rhizobia. This study clarified that R. anhuiense was the main symbiont for beach pea rhizobia on the seashore line of Shandong Peninsula. The low level genetic diversity of beach pea rhizobia revealed by both MLSA and the symbiotic genes might be related to the strong selection pressure produced by the saline-alkaline environment and the host plants. Copyright © 2016 Elsevier GmbH. All rights reserved.

  8. Avaliação da reação de mitsuda em pacientes virchovianos inativos antes e após imunoterapia

    Directory of Open Access Journals (Sweden)

    Maria Sueli Parreira de Arruda

    1995-09-01

    Full Text Available Neste estudo investigou-se o potencial imunomodulador do levamisole e da mistura BCG/Mycobacterium leprae em pacientes virchovianos inativos, utilizando como parâmetro a reação de Mitsuda. Vinte pacientes, classificados como Mitsuda histologicamente negativos há 10 anos, foram divididos em três grupos: cinco pacientes que foram somente reavaliados frente a mitsudina: oito pacientes que receberam levamisole e, sete que receberam a mistura de BCG vivo mais M. leprae morto. Os resultados mostraram que: 1 o levamisole não alterou a reatividade à mitsudina em nenhum dos casos estudados; 2 as modificações da reatividade verificadas com o uso da mistura (tres casos ou aquelas que ocorreram espontaneamente (tres casos foram sempre de pequena amplitude e refletiram variações próprias de pacientes com algum grau de resistência ao Mycobacterium leprae.In this study the immunopotentiator levamisole as well as a mixture of BCGMycobacterium leprae were investigated in inactive lepromatous leprosy patients by using the Mitsuda reaction as a parameter. Twenty lepromatous patients ten years ago classified as histologically negative for Mitsuda's test were divided into three groups: five patients that were only retested with Mitsuda antigen; eight patients that received oral levamisol and seven patients that received a mixture of alive BCG plus autoclaved M. leprae.The results indicated that: 1 the levamisole did not alter the reactivity to lepromin in any of the patients studied 2 neither the changes in the reactivity to lepromin by using the mixture (3 cases nor those that occurred spontaneously (3 cases were clear. They properly reflected the natural variation of patients with some degree of resistance to Mycobacterium leprae.

  9. A migration-driven model for the historical spread of leprosy in medieval Eastern and Central Europe.

    Science.gov (United States)

    Donoghue, Helen D; Michael Taylor, G; Marcsik, Antónia; Molnár, Erika; Pálfi, Gyorgy; Pap, Ildikó; Teschler-Nicola, Maria; Pinhasi, Ron; Erdal, Yilmaz S; Velemínsky, Petr; Likovsky, Jakub; Belcastro, Maria Giovanna; Mariotti, Valentina; Riga, Alessandro; Rubini, Mauro; Zaio, Paola; Besra, Gurdyal S; Lee, Oona Y-C; Wu, Houdini H T; Minnikin, David E; Bull, Ian D; O'Grady, Justin; Spigelman, Mark

    2015-04-01

    Leprosy was rare in Europe during the Roman period, yet its prevalence increased dramatically in medieval times. We examined human remains, with paleopathological lesions indicative of leprosy, dated to the 6th-11th century AD, from Central and Eastern Europe and Byzantine Anatolia. Analysis of ancient DNA and bacterial cell wall lipid biomarkers revealed Mycobacterium leprae in skeletal remains from 6th-8th century Northern Italy, 7th-11th century Hungary, 8th-9th century Austria, the Slavic Greater Moravian Empire of the 9th-10th century and 8th-10th century Byzantine samples from Northern Anatolia. These data were analyzed alongside findings published by others. M. leprae is an obligate human pathogen that has undergone an evolutionary bottleneck followed by clonal expansion. Therefore M. leprae genotypes and sub-genotypes give information about the human populations they have infected and their migration. Although data are limited, genotyping demonstrates that historical M. leprae from Byzantine Anatolia, Eastern and Central Europe resembles modern strains in Asia Minor rather than the recently characterized historical strains from North West Europe. The westward migration of peoples from Central Asia in the first millennium may have introduced different M. leprae strains into medieval Europe and certainly would have facilitated the spread of any existing leprosy. The subsequent decline of M. leprae in Europe may be due to increased host resistance. However, molecular evidence of historical leprosy and tuberculosis co-infections suggests that death from tuberculosis in leprosy patients was also a factor. Copyright © 2015 Elsevier B.V. All rights reserved.

  10. Current strategy for leprosy control in Brazil: time to pursue alternative preventive strategies?

    OpenAIRE

    Sérgio S. Cunha; Laura C. Rodrigues; Nádia Cristina Duppre

    2004-01-01

    La estrategia actual para el control de la lepra en Brasil se basa en dos actividades principales: la detección precoz de casos y el tratamiento de casos con farmacoterapia combinada. Además de dichas medidas, se realizan esfuerzos complementarios para identificar los contactos domésticos para el diagnóstico precoz y la vacunación con el bacilo de Calmette-Guérin (BCG). Sin embargo, la eficacia de estas acciones a la hora de reducir la incidencia de la lepra es aún discutible. Esto genera dud...

  11. Antioxidant factors, nitric oxide levels, and cellular damage in leprosy patients

    Directory of Open Access Journals (Sweden)

    Taysa Ribeiro Schalcher

    2013-09-01

    Full Text Available Introduction The immune response caused by Mycobacterium leprae is a risk factor for the development of oxidative stress (OS in leprosy patients. This study aimed to assess OS in leprosy patients before the use of a multidrug therapy. Methods We evaluated the nitric oxide (NO concentration; antioxidant capacity; levels of malondialdehyde, methemoglobin and reduced glutathione; and the activity of catalase and superoxide dismutase (SOD in leprosy patients. Results We observed lower SOD activity in these leprosy patients; however, the NO levels and antioxidant capacity were increased. Conclusions The infectious process in response to M. leprae could primarily be responsible for the OS observed in these patients.

  12. Mutations for Worse or Better: Low-Fidelity DNA Synthesis by SOS DNA Polymerase V Is a Tightly Regulated Double-Edged Sword.

    Science.gov (United States)

    Jaszczur, Malgorzata; Bertram, Jeffrey G; Robinson, Andrew; van Oijen, Antoine M; Woodgate, Roger; Cox, Michael M; Goodman, Myron F

    2016-04-26

    1953, the year of Watson and Crick, bore witness to a less acclaimed yet highly influential discovery. Jean Weigle demonstrated that upon infection of Escherichia coli, λ phage deactivated by UV radiation, and thus unable to form progeny, could be reactivated by irradiation of the bacterial host. Evelyn Witkin and Miroslav Radman later revealed the presence of the SOS regulon. The more than 40 regulon genes are repressed by LexA protein and induced by the coproteolytic cleavage of LexA, catalyzed by RecA protein bound to single-stranded DNA, the RecA* nucleoprotein filament. Several SOS-induced proteins are engaged in repairing both cellular and extracellular damaged DNA. There's no "free lunch", however, because error-free repair is accompanied by error-prone translesion DNA synthesis (TLS), involving E. coli DNA polymerase V (UmuD'2C) and RecA*. This review describes the biochemical mechanisms of pol V-mediated TLS. pol V is active only as a mutasomal complex, pol V Mut = UmuD'2C-RecA-ATP. RecA* donates a single RecA subunit to pol V. We highlight three recent insights. (1) pol V Mut has an intrinsic DNA-dependent ATPase activity that governs polymerase binding and dissociation from DNA. (2) Active and inactive states of pol V Mut are determined at least in part by the distinct interactions between RecA and UmuC. (3) pol V is activated by RecA*, not at a blocked replisome, but at the inner cell membrane.

  13. Comparative Genomics of Chrysochromulina Ericina Virus and Other Microalga-Infecting Large DNA Viruses Highlights Their Intricate Evolutionary Relationship with the Established Mimiviridae Family.

    Science.gov (United States)

    Gallot-Lavallée, Lucie; Blanc, Guillaume; Claverie, Jean-Michel

    2017-07-15

    Chrysochromulina ericina virus CeV-01B (CeV) was isolated from Norwegian coastal waters in 1998. Its icosahedral particle is 160 nm in diameter and encloses a 474-kb double-stranded DNA (dsDNA) genome. This virus, although infecting a microalga (the haptophyceae Haptolina ericina , formerly Chrysochromulina ericina ), is phylogenetically related to members of the Mimiviridae family, initially established with the acanthamoeba-infecting mimivirus and megavirus as prototypes. This family was later split into two genera ( Mimivirus and Cafeteriavirus ) following the characterization of a virus infecting the heterotrophic stramenopile Cafeteria roenbergensis (CroV). CeV, as well as two of its close relatives, which infect the unicellular photosynthetic eukaryotes Phaeocystis globosa (Phaeocystis globosa virus [PgV]) and Aureococcus anophagefferens (Aureococcus anophagefferens virus [AaV]), are currently unclassified by the International Committee on Viral Taxonomy (ICTV). The detailed comparative analysis of the CeV genome presented here confirms the phylogenetic affinity of this emerging group of microalga-infecting viruses with the Mimiviridae but argues in favor of their classification inside a distinct clade within the family. Although CeV, PgV, and AaV share more common features among them than with the larger Mimiviridae , they also exhibit a large complement of unique genes, attesting to their complex evolutionary history. We identified several gene fusion events and cases of convergent evolution involving independent lateral gene acquisitions. Finally, CeV possesses an unusual number of inteins, some of which are closely related despite being inserted in nonhomologous genes. This appears to contradict the paradigm of allele-specific inteins and suggests that the Mimiviridae are especially efficient in spreading inteins while enlarging their repertoire of homing genes. IMPORTANCE Although it infects the microalga Chrysochromulina ericina , CeV is more closely

  14. Segmental isotopic labeling by asparaginyl endopeptidase-mediated protein ligation.

    Science.gov (United States)

    Mikula, Kornelia M; Krumwiede, Luisa; Plückthun, Andreas; Iwaï, Hideo

    2018-03-13

    Segmental isotopic labeling can facilitate NMR studies of large proteins, multi-domain proteins, and proteins with repetitive sequences by alleviating NMR signal overlaps. Segmental isotopic labeling also allows us to investigate an individual domain in the context of a full-length protein by NMR. Several established methods are available for segmental isotopic labeling such as intein-mediated ligation, but each has specific requirements and limitations. Here, we report an enzymatic approach using bacterially produced asparagine endopeptidase from Oldenlandia affinis for segmental isotopic labeling of a protein with repetitive sequences, a designed armadillo repeat protein, by overcoming some of the shortcomings of enzymatic ligation for segmental isotopic labeling.

  15. SICLOPPS cyclic peptide libraries in drug discovery.

    Science.gov (United States)

    Tavassoli, Ali

    2017-06-01

    Cyclic peptide libraries have demonstrated significant potential when employed against challenging targets such as protein-protein interactions. While a variety of methods for library generation exist, genetically encoded libraries hold several advantages over their chemically synthesized counterparts; they are more readily accessible and allow straightforward hit deconvolution. One method for the intracellular generation of such libraries is split-intein circular ligation of peptides and proteins (SICLOPPS). Here we detail and discuss the deployment of SICLOPPS libraries for the identification of cyclic peptide inhibitors of a variety of targets. Copyright © 2017 Elsevier Ltd. All rights reserved.

  16. ATP synthase from slow and fast growing mycobacteria is active in ATP synthesis and blocked in ATP hydrolysis direction.

    NARCIS (Netherlands)

    Haagsma, A.C.; Driessen, N.N.; Hahn, M.M.; Lill, H.; Bald, D.

    2010-01-01

    ATP synthase is a validated drug target for the treatment of tuberculosis, and ATP synthase inhibitors are promising candidate drugs for the treatment of infections caused by other slow-growing mycobacteria, such as Mycobacterium leprae and Mycobacterium ulcerans. ATP synthase is an essential enzyme

  17. ATP synthase in slow- and fast-growing mycobacteria is active in ATP synthesis and blocked in ATP hydrolysis direction.

    NARCIS (Netherlands)

    Haagsma, A.C.; Driessen, N.N.; Hahn, M.M.; Lill, H.; Bald, D.

    2010-01-01

    ATP synthase is a validated drug target for the treatment of tuberculosis, and ATP synthase inhibitors are promising candidate drugs for the treatment of infections caused by other slow-growing mycobacteria, such as Mycobacterium leprae and Mycobacterium ulcerans. ATP synthase is an essential enzyme

  18. Servicio de noticias médicas y farmacéuticas

    Directory of Open Access Journals (Sweden)

    Facultad de Medicina Revista

    1955-09-01

    Full Text Available Nuevo Tratamiento de la Epilepsia; La Atropina impide el desenlace fatal en la intoxicación letal por Insecticidas a base de Fósforo; Curación y Control de la Lepra; Tratamiento de las Infecciones de las Vías Urinarias; Rehabilitación de los Enfermos Cardíacos.

  19. Understanding the stigma of leprosy | Luka | South Sudan Medical ...

    African Journals Online (AJOL)

    Leprosy is the oldest disease known to man. The earliest written records describing true leprosy came from India around the period 600 BC. Leprosy is caused by Mycobacterium leprae; the Norwegian, Dr Amauer Hansen, isolated the bacterium in 1873. Leprosy is also called Hansen's disease after him. Although it is the ...

  20. ARC: Automated Resource Classifier for agglomerative functional ...

    Indian Academy of Sciences (India)

    PRAKASH KUMAR

    Colibri], TubercuList. (Mycobacterium tuberculosis H37Rv [TubercuList],. Leproma (Mycobacterium leprae TN) [Leproma], BoviList. (Mycobacterium bovis AF2122/97) [BoviList], ListiList. (Listeria monocytogenes EGD-e, L. innocua CLIP 11262).

  1. A BACTERIAL CYTOKINE Mukamolova et al (1998) PNAS, 95, 8916 ...

    Indian Academy of Sciences (India)

    First page Back Continue Last page Overview Graphics. A BACTERIAL CYTOKINE Mukamolova et al (1998) PNAS, 95, 8916-8921. To grow or not to grow is a social decision. A resuscitation promoting factor (rpf) identified. The rpf protein shares similarity with M. tuberculosis and M. leprae ...

  2. A migration-driven model for the historical spread of leprosy in medieval Eastern and Central Europe

    Czech Academy of Sciences Publication Activity Database

    Donoghue, H. D.; Taylor, G. M.; Marcsik, A.; Molnár, E.; Pálfi, G.; Pap, I.; Teschler-Nicola, M.; Pinhasi, R.; Erdal, Y. S.; Velemínský, P.; Likovský, Jakub; Belcastro, M. G.; Mariotti, V.; Riga, A.; Rubini, M.; Zaio, P.; Besra, G. S.; Lee, O. Y.-C.; Wu, H. H.T.; Minnikin, D. E.; Bull, I. D.; O'Grady, J.; Spigelman, M.

    2015-01-01

    Roč. 31, April (2015), s. 250-256 ISSN 1567-1348 Institutional support: RVO:67985912 Keywords : ancient DNA * genotyping * human migrations * lipid biomarkers * Mycobacterium leprae * Mycobacterium tuberculosis Subject RIV: AC - Archeology, Anthropology, Ethnology Impact factor: 2.591, year: 2015

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

    OpenAIRE

    Young, Douglas B.; Comas, I?aki; de Carvalho, Luiz P. S.

    2015-01-01

    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 (SNPs) with predicted impact on protein function and transcriptional regulation...

  4. Utility of Bacillary Index in Slit Skin Smears in Correlation with ...

    African Journals Online (AJOL)

    Mycobacterium leprae not only affects skin and peripheral nerves but also involves muscles, eyes, bones, testis, and internal organs.[1]. Leprosy is a disease bedeviled by many classifications throughout history – Madrid classification (ILC 1953), Ridley and Jopling classification (1966), and Indian classification. (IAL 1982).

  5. Fulltext PDF

    Indian Academy of Sciences (India)

    Admin

    Tuberculosis got its present name at the end of the. 19th century. Under its previous name of consumption it has been long known as a world- wide phenomenon. It is caused by a bacterium, Mycobacterium tuberculosis. A related organism, M. bovis, causes TB in cattle while M. leprae is the causative agent of leprosy in man ...

  6. The Greek moulages: a picture of skin diseases in former times.

    Science.gov (United States)

    Worm, A-M; Hadjivassiliou, M; Katsambas, A

    2007-04-01

    Medical moulages are three-dimensional wax figures made for teaching in the beginning of the last century. A rather unknown hospital museum in Athens, Greece, stores 1660 moulages depicting skin and venereal diseases prevalent at that time such as syphilis, lepra and tuberculosis. The historical background behind the Greek moulages and the art of moulaging are described.

  7. African Journal of Infectious Diseases - Vol 12, No 1S (2018)

    African Journals Online (AJOL)

    Acanthamoeba Sp. S-11 phagocytotic activity on Mycobacterium leprae in different nutrient conditions · EMAIL FREE FULL TEXT EMAIL FREE FULL TEXT DOWNLOAD FULL TEXT DOWNLOAD FULL TEXT. Sepling Paling, Ratna Wahyuni, Ni'matuzahroh Ni'matuzahroh, Dwi Winarni, Iswahyudi Iswahyudi, Linda Astari, ...

  8. [Leprosy, a pillar of human genetics of infectious diseases].

    Science.gov (United States)

    Gaschignard, J; Scurr, E; Alcaïs, A

    2013-06-01

    Despite a natural reservoir of Mycobacterium leprae limited to humans and free availability of an effective antibiotic treatment, more than 200,000 people develop leprosy each year. This disease remains a major cause of disability and social stigma worldwide. The cause of this constant incidence is currently unknown and indicates that important aspects of the complex relationship between the pathogen and its human host remain to be discovered. An important contribution of host genetics to susceptibility to leprosy has long been suggested to account for the considerable variability between individuals sustainably exposed to M. leprae. Given the inability to cultivate M. leprae in vitro and in the absence of relevant animal model, genetic epidemiology is the main strategy used to identify the genes and, consequently, the immunological pathways involved in protective immunity to M. leprae. Recent genome-wide studies have identified new pathophysiological pathways which importance is only beginning to be understood. In addition, the prism of human genetics placed leprosy at the crossroads of other common diseases such as Crohn's disease, asthma or myocardial infarction. Therefore, novel lights on the pathogenesis of many common diseases could eventually emerge from the detailed understanding of a disease of the shadows. Copyright © 2013 Elsevier Masson SAS. All rights reserved.

  9. Immunohistochemical detection of PGL-1, LAM, 30 kD and 65 kD antigens in leprosy infected paraffin preserved skin and nerve sections

    NARCIS (Netherlands)

    van den Bos, I. C.; Khanolkar-Young, S.; Das, P. K.; Lockwood, D. N.

    1999-01-01

    A panel of lipid, carbohydrate and protein antibodies were optimized for use in detecting M. leprae antigens in paraffin embedded material. Skin and nerve biopsies from 13 patients across the leprosy spectrum were studied. All antibodies detected antigen in tissues with a BI > 1. Phenolic-glycolipid

  10. Utility of measuring serum levels of anti-PGL-I antibody, neopterin and C-reactive protein in monitoring leprosy patients during multi-drug treatment and reactions

    NARCIS (Netherlands)

    Silva, E. A.; Iyer, A.; Ura, S.; Lauris, J. R.; Naafs, B.; Das, P. K.; Vilani-Moreno, F.

    2007-01-01

    To verify the validity of measuring the levels of Mycobacterium leprae-specific anti-phenolic glycolipid (PGL)-I antibody, neopterin, a product of activated macrophages, and C-reactive protein (CRP), an acute phase protein, in serial serum samples from patients for monitoring the leprosy spectrum

  11. The armadillo: a model for the neuropathy of leprosy and potentially other neurodegenerative diseases

    Directory of Open Access Journals (Sweden)

    Rahul Sharma

    2013-01-01

    Full Text Available Leprosy (also known as Hansen’s disease is an infectious peripheral neurological disorder caused by Mycobacterium leprae that even today leaves millions of individuals worldwide with life-long disabilities. The specific mechanisms by which this bacterium induces nerve injury remain largely unknown, mainly owing to ethical and practical limitations in obtaining affected human nerve samples. In addition to humans, nine-banded armadillos (Dasypus novemcinctus are the only other natural host of M. leprae, and they develop a systemically disseminated disease with extensive neurological involvement. M. leprae is an obligate intracellular parasite that cannot be cultivated in vitro. Because of the heavy burdens of bacilli they harbor, nine-banded armadillos have become the organism of choice for propagating large quantities of M. leprae, and they are now advancing as models of leprosy pathogenesis and nerve damage. Although armadillos are exotic laboratory animals, the recently completed whole genome sequence for this animal is enabling researchers to undertake more sophisticated molecular studies and to develop armadillo-specific reagents. These advances will facilitate the use of armadillos in piloting new therapies and diagnostic regimens, and will provide new insights into the oldest known infectious neurodegenerative disorder.

  12. Eritema nudoso leproso persistente y enteropatía letal por clofazimina

    Directory of Open Access Journals (Sweden)

    Gerzaín Rodríguez

    2009-03-01

    Conclusiones. La clínica final de la paciente sugiere que se trata de un caso de enteropatía letal por clofazimina, una complicación que no se había reconocido previamente en nuestros pacientes. Es necesario aumentar el conocimiento de la lepra entre los médicos.

  13. Validation of qPCR Methods for the Detection of Mycobacterium in New World Animal Reservoirs.

    Directory of Open Access Journals (Sweden)

    Genevieve Housman

    2015-11-01

    Full Text Available Zoonotic pathogens that cause leprosy (Mycobacterium leprae and tuberculosis (Mycobacterium tuberculosis complex, MTBC continue to impact modern human populations. Therefore, methods able to survey mycobacterial infection in potential animal hosts are necessary for proper evaluation of human exposure threats. Here we tested for mycobacterial-specific single- and multi-copy loci using qPCR. In a trial study in which armadillos were artificially infected with M. leprae, these techniques were specific and sensitive to pathogen detection, while more traditional ELISAs were only specific. These assays were then employed in a case study to detect M. leprae as well as MTBC in wild marmosets. All marmosets were negative for M. leprae DNA, but 14 were positive for the mycobacterial rpoB gene assay. Targeted capture and sequencing of rpoB and other MTBC genes validated the presence of mycobacterial DNA in these samples and revealed that qPCR is useful for identifying mycobacterial-infected animal hosts.

  14. Leprosy Associated with Atypical Cutaneous Leishmaniasis in Nicaragua and Honduras.

    Science.gov (United States)

    Soto, Lucrecia Acosta; Caballero, Nelson; Fuentes, Lesny Ruth; Muñoz, Pedro Torres; Gómez Echevarría, Jose Ramón; López, Montserrat Pérez; Bornay Llinares, Fernando Jorge; Stanford, John L; Stanford, Cynthia A; Donoghue, Helen D

    2017-10-01

    In Central America, few cases of leprosy have been reported, but the disease may be unrecognized. Diagnosis is based on clinical criteria and histology. Preliminary field work in Nicaragua and Honduras found patients, including many children, with skin lesions clinically suggestive of atypical cutaneous leishmaniasis or indeterminate leprosy. Histology could not distinguish these diseases although acid-fast organisms were visible in a few biopsies. Lesions healed after standard antimicrobial therapy for leprosy. In the present study, patients, family members, and other community members were skin-tested and provided nasal swabs and blood samples. Biopsies were taken from a subgroup of patients with clinical signs of infection. Two laboratories analyzed samples, using local in-house techniques. Mycobacterium leprae , Leishmania spp. and Leishmania infantum were detected using polymerase chain reactions. Mycobacterium leprae DNA was detected in blood samples and nasal swabs, including some cases where leprosy was not clinically suspected. Leishmania spp. were also detected in blood and nasal swabs. Most biopsies contained Leishmania DNA and coinfection of Leishmania spp. with M. leprae occurred in 33% of cases. Mycobacterium leprae DNA was also detected and sequenced from Nicaraguan and Honduran environmental samples. In conclusion, leprosy and leishmaniasis are present in both regions, and leprosy appears to be widespread. The nature of any relationship between these two pathogens and the epidemiology of these infections need to be elucidated.

  15. Author Details

    African Journals Online (AJOL)

    Acanthamoeba Sp. S-11 phagocytotic activity on Mycobacterium leprae in different nutrient conditions. Abstract PDF. ISSN: 2006-0165. AJOL African Journals Online. HOW TO USE AJOL... for Researchers · for Librarians · for Authors · FAQ's · More about AJOL · AJOL's Partners · Terms and Conditions of Use · Contact AJOL ...

  16. Creating Directed Double-strand Breaks with the Ref Protein: A Novel Rec A-Dependent Nuclease from Bacteriophage P1

    Energy Technology Data Exchange (ETDEWEB)

    Gruenig, Marielle C.; Lu, Duo; Won, Sang Joon; Dulberger, Charles L.; Manlick, Angela J.; Keck, James L.; Cox, Michael M. (UW)

    2012-03-16

    The bacteriophage P1-encoded Ref protein enhances RecA-dependent recombination in vivo by an unknown mechanism. We demonstrate that Ref is a new type of enzyme; that is, a RecA-dependent nuclease. Ref binds to ss- and dsDNA but does not cleave any DNA substrate until RecA protein and ATP are added to form RecA nucleoprotein filaments. Ref cleaves only where RecA protein is bound. RecA functions as a co-nuclease in the Ref/RecA system. Ref nuclease activity can be limited to the targeted strands of short RecA-containing D-loops. The result is a uniquely programmable endonuclease activity, producing targeted double-strand breaks at any chosen DNA sequence in an oligonucleotide-directed fashion. We present evidence indicating that cleavage occurs in the RecA filament groove. The structure of the Ref protein has been determined to 1.4 {angstrom} resolution. The core structure, consisting of residues 77-186, consists of a central 2-stranded {beta}-hairpin that is sandwiched between several {alpha}-helical and extended loop elements. The N-terminal 76 amino acid residues are disordered; this flexible region is required for optimal activity. The overall structure of Ref, including several putative active site histidine residues, defines a new subclass of HNH-family nucleases. We propose that enhancement of recombination by Ref reflects the introduction of directed, recombinogenic double-strand breaks.

  17. The study of the extreme radiation tolerance mechanisms of the bacterium Deinococcus deserti by a functional genomics approach

    International Nuclear Information System (INIS)

    Dulermo, R.

    2009-12-01

    The genome of Deinococcus deserti, a highly radiation-tolerant bacterium, was analyzed and compared to those of D. radiodurans and D. geothermalis. About 230 proteins are specifically conserved in these 3 species, including IrrE, a regulator protein essential for radio tolerance. D.deserti has several supplementary DNA repair genes, like imuY and dnaE2 (trans-lesion DNA polymerases). Moreover, D. deserti has 3 recA that code for 2 different RecA proteins (RecAC et RecAP). To study these genes, genetic tools were developed for D. deserti. Different results suggest that IrrE, required for the induction of several genes after irradiation, has peptidase activity. The 2 RecA proteins are functional for DNA repair. D. deserti is mutable by UV, which requires ImuY, DnaE2 and RecAC, but not RecAP. (author)

  18. Genetic control of near-UV sensitivity independent of excision deficiency (uvrA6) in Escherichia coli K12

    International Nuclear Information System (INIS)

    Tuveson, R.W.

    1980-01-01

    By appropriate matings, recombinant strains carrying all four possible combinations of genes controlling near-UV (nur vs nur + ) and far-UV (uvrA6 vs uvrA + , excision repair function) sensitivity were constructed. Near and far-UV inactivation experiments with the four recombinant strains revealed that inactivating events induced by near and far-UV do not appear to overlap. These results are analogous to previously reported experiments with recombinant strains carrying all four possible combinations of genes controlling near-UV sensitivity (nur vs nur + ) and recombination proficiency (far-UV sensitivity, recA1 vs recA + ). The results of these two sets of experiments taken together may mean that any recA + or uvrA + repairable lesions induced by near-UV are repaired equally well by either system and do not require the simultaneous participation of both repair systems. (author)

  19. recA-based PCR assay for accurate differentiation of Streptococcus pneumoniae from other viridans streptococci.

    Science.gov (United States)

    Zbinden, A; Köhler, N; Bloemberg, G V

    2011-02-01

    Proper identification of Streptococcus pneumoniae by conventional methods remains problematic. The discriminatory power of the 16S rRNA gene, which can be considered the "gold standard" for molecular identification, is too low to differentiate S. pneumoniae from closely related species such as Streptococcus pseudopneumoniae, Streptococcus mitis, and Streptococcus oralis in the routine clinical laboratory. A 313-bp part of recA was selected on the basis of variability within the S. mitis group, showing <95.8% interspecies homology. In addition, 6 signature nucleotides specific for S. pneumoniae were identified within the 313-bp recA fragment. We show that recA analysis is a useful tool for proper identification to species level within the S. mitis group, in particular, for pneumococci.

  20. recA-Based PCR Assay for Accurate Differentiation of Streptococcus pneumoniae from Other Viridans Streptococci▿

    Science.gov (United States)

    Zbinden, A.; Köhler, N.; Bloemberg, G. V.

    2011-01-01

    Proper identification of Streptococcus pneumoniae by conventional methods remains problematic. The discriminatory power of the 16S rRNA gene, which can be considered the “gold standard” for molecular identification, is too low to differentiate S. pneumoniae from closely related species such as Streptococcus pseudopneumoniae, Streptococcus mitis, and Streptococcus oralis in the routine clinical laboratory. A 313-bp part of recA was selected on the basis of variability within the S. mitis group, showing <95.8% interspecies homology. In addition, 6 signature nucleotides specific for S. pneumoniae were identified within the 313-bp recA fragment. We show that recA analysis is a useful tool for proper identification to species level within the S. mitis group, in particular, for pneumococci. PMID:21147955

  1. Bacterial inclusion bodies as potential synthetic devices for pathogen recognition and a therapeutic substance release.

    Science.gov (United States)

    Talafová, Klaudia; Hrabárová, Eva; Chorvát, Dušan; Nahálka, Jozef

    2013-02-07

    Adhesins of pathogens recognise the glycans on the host cell and mediate adherence. They are also crucial for determining the tissue preferences of pathogens. Currently, glyco-nanomaterials provide potential tool for antimicrobial therapy. We demonstrate that properly glyco-tailored inclusion bodies can specifically bind pathogen adhesins and release therapeutic substances. In this paper, we describe the preparation of tailored inclusion bodies via the conjugation of indicator protein aggregated to form inclusion bodies with soluble proteins. Whereas the indicator protein represents a remedy, the soluble proteins play a role in pathogen recognition. For conjugation, glutaraldehyde was used as linker. The treatment of conjugates with polar lysine, which was used to inactivate the residual glutaraldehyde, inhibited unwanted hydrophobic interactions between inclusion bodies. The tailored inclusion bodies specifically interacted with the SabA adhesin from Helicobacter pylori aggregated to form inclusion bodies that were bound to the sialic acids decorating the surface of human erythrocytes. We also tested the release of indicator proteins from the inclusion bodies using sortase A and Ssp DNAB intein self-cleaving modules, respectively. Sortase A released proteins in a relatively short period of time, whereas the intein cleavage took several weeks. The tailored inclusion bodies are promising "nanopills" for biomedical applications. They are able to specifically target the pathogen, while a self-cleaving module releases a soluble remedy. Various self-cleaving modules can be enabled to achieve the diverse pace of remedy release.

  2. Split2 Protein-Ligation Generates Active IL-6-Type Hyper-Cytokines from Inactive Precursors.

    Science.gov (United States)

    Moll, Jens M; Wehmöller, Melanie; Frank, Nils C; Homey, Lisa; Baran, Paul; Garbers, Christoph; Lamertz, Larissa; Axelrod, Jonathan H; Galun, Eithan; Mootz, Henning D; Scheller, Jürgen

    2017-12-15

    Trans-signaling of the major pro- and anti-inflammatory cytokines Interleukin (IL)-6 and IL-11 has the unique feature to virtually activate all cells of the body and is critically involved in chronic inflammation and regeneration. Hyper-IL-6 and Hyper-IL-11 are single chain designer trans-signaling cytokines, in which the cytokine and soluble receptor units are trapped in one complex via a flexible peptide linker. Albeit, Hyper-cytokines are essential tools to study trans-signaling in vitro and in vivo, the superior potency of these designer cytokines are accompanied by undesirable stress responses. To enable tailor-made generation of Hyper-cytokines, we developed inactive split-cytokine-precursors adapted for posttranslational reassembly by split-intein mediated protein trans-splicing (PTS). We identified cutting sites within IL-6 (E 134 /S 135 ) and IL-11 (G 116 /S 117 ) and obtained inactive split-Hyper-IL-6 and split-Hyper-IL-11 cytokine precursors. After fusion with split-inteins, PTS resulted in reconstitution of active Hyper-cytokines, which were efficiently secreted from transfected cells. Our strategy comprises the development of a background-free cytokine signaling system from reversibly inactivated precursor cytokines.

  3. 3′-Terminated Overhangs Regulate DNA Double-Strand Break Processing in Escherichia coli

    Directory of Open Access Journals (Sweden)

    Edyta Đermić

    2017-09-01

    Full Text Available Double-strand breaks (DSBs are lethal DNA lesions, which are repaired by homologous recombination in Escherichia coli. To study DSB processing in vivo, we induced DSBs into the E. coli chromosome by γ-irradiation and measured chromosomal degradation. We show that the DNA degradation is regulated by RecA protein concentration and its rate of association with single-stranded DNA (ssDNA. RecA decreased DNA degradation in wild-type, recB, and recD strains, indicating that it is a general phenomenon in E. coli. On the other hand, DNA degradation was greatly reduced and unaffected by RecA in the recB1080 mutant (which produces long overhangs and in a strain devoid of four exonucleases that degrade a 3′ tail (ssExos. 3′–5′ ssExos deficiency is epistatic to RecA deficiency concerning DNA degradation, suggesting that bound RecA is shielding the 3′ tail from degradation by 3′–5′ ssExos. Since 3′ tail preservation is common to all these situations, we infer that RecA polymerization constitutes a subset of mechanisms for preserving the integrity of 3′ tails emanating from DSBs, along with 3′ tail’s massive length, or prevention of their degradation by inactivation of 3′–5′ ssExos. Thus, we conclude that 3′ overhangs are crucial in controlling the extent of DSB processing in E. coli. This study suggests a regulatory mechanism for DSB processing in E. coli, wherein 3′ tails impose a negative feedback loop on DSB processing reactions, specifically on helicase reloading onto dsDNA ends.

  4. Analysis of spontaneous deletions and gene amplification in the lac region of Escherichia coli

    International Nuclear Information System (INIS)

    Albertini, A.M.; Hofer, M.; Calos, M.P.; Tlsty, T.D.; Miller, J.H.

    1983-01-01

    Spontaneous rearrangements, such as large deletions and duplications, have important implications for the structure of the genome. It is therefore of great interest to analyze these events at the molecular level. We have constructed derivatives of a lacI-Z fusion strain, which allow us to study deletions in a more systematic manner than was previously possible. These derivatives have been used to investigate how frequently larger deletions (> 700 bp) occur between short homologies on both recA and recA - strains and to determine the effect of the lengths of the short homologies and of the distance between homologies on the frequency of deletion formation. 38 references, 11 figures

  5. Role of the RecF gene product in UV mutagenesis of lambda phage

    International Nuclear Information System (INIS)

    Wood, R.D.; Stein, J.

    1986-01-01

    E. coli recF mutants have a greatly reduced capacity for Weigle mutagenesis of ultraviolet light-irradiated lambda phage. A recF 332::Tn3 mutation was introduced into an E. coli recA441 lex A51 strain which constitutively expresses SOS functions. Weigle mutagenesis of phage lambda could occur in the resulting strain in the absence of host cell irradiation, and was increased when the recA441 (tif) allele was activated of recF strains to support Weigle mutagenesis can therefore be ascribed to a defect in expression of SOS functions after irradiation. (orig.)

  6. Effect of the psi B gene on the indirect recombinogenesis of the lambda bacteriophage; Efecto del gen psi B sobre la recombinogenesis indirecta del bacteriofago lambda

    Energy Technology Data Exchange (ETDEWEB)

    Alcantara D, D. [ININ, 52045 Ocoyoacac, Estado de Mexico (Mexico)

    1993-10-15

    Presently work, we prove if the protein psi B suppresses the indirect recombinogenesis of lambda when inhibiting the induction of the system bacterial SOS. This experimental model's advantage is that it allows to exclude the activity of co protease of RecA selectively without affecting the activity of recombinases of the same one, making possible the analysis of the paper that play both functions in the phenomenon. The results show that the inhibition of the activity of co protease of RecA doesn't suppress the indirect recombinogenesis of lambda. (Author)

  7. Effect of the psi B gene on the indirect recombinogenesis of the lambda bacteriophage

    International Nuclear Information System (INIS)

    Alcantara D, D.

    1993-10-01

    Presently work, we prove if the protein psi B suppresses the indirect recombinogenesis of lambda when inhibiting the induction of the system bacterial SOS. This experimental model's advantage is that it allows to exclude the activity of co protease of RecA selectively without affecting the activity of recombinases of the same one, making possible the analysis of the paper that play both functions in the phenomenon. The results show that the inhibition of the activity of co protease of RecA doesn't suppress the indirect recombinogenesis of lambda. (Author)

  8. Effects of radiations on DNA and repair of the damage. Progress report, March 1, 1975--March 31, 1976

    International Nuclear Information System (INIS)

    Hutchinson, F.

    1976-01-01

    It was established that repair of radioinduced double-strand breaks in the DNA of E. coli AB2497 takes place. This repair can be eliminated by growing the cells in poor media so there is only 1+ genome/cell. There is no measurable repair in AB2487 recA - (otherwise isogenic with AB2497) or NH4803 recA - recB - cells. These results strongly suggest that DNA double-strand break repair occurs by a process involving recombination of the broken pieces with a homologous double hexix

  9. Structural basis for the function of DEAH helicases

    DEFF Research Database (Denmark)

    He, Yangzi; Andersen, Gregers Rom; Nielsen, Klaus Hvid

    2010-01-01

    DEAH helicases participate in pre‐messenger RNA splicing and ribosome biogenesis. The structure of yeast Prp43p‐ADP reveals the homology of DEAH helicases to DNA helicases and the presence of an oligonucleotide‐binding motif. A β‐hairpin from the second RecA domain is wedged between two carboxy......‐terminal domains and blocks access to the occluded RNA binding site formed by the RecA domains and a C‐terminal domain. ATP binding and hydrolysis are likely to induce conformational changes in the hairpin that are important for RNA unwinding or ribonucleoprotein remodelling. The structure of Prp43p provides...

  10. Single Strand Annealing Plays a Major Role in RecA-Independent Recombination between Repeated Sequences in the Radioresistant Deinococcus radiodurans Bacterium.

    Directory of Open Access Journals (Sweden)

    Solenne Ithurbide

    2015-10-01

    Full Text Available The bacterium Deinococcus radiodurans is one of the most radioresistant organisms known. It is able to reconstruct a functional genome from hundreds of radiation-induced chromosomal fragments. Our work aims to highlight the genes involved in recombination between 438 bp direct repeats separated by intervening sequences of various lengths ranging from 1,479 bp to 10,500 bp to restore a functional tetA gene in the presence or absence of radiation-induced DNA double strand breaks. The frequency of spontaneous deletion events between the chromosomal direct repeats were the same in recA+ and in ΔrecA, ΔrecF, and ΔrecO bacteria, whereas recombination between chromosomal and plasmid DNA was shown to be strictly dependent on the RecA and RecF proteins. The presence of mutations in one of the repeated sequence reduced, in a MutS-dependent manner, the frequency of the deletion events. The distance between the repeats did not influence the frequencies of deletion events in recA+ as well in ΔrecA bacteria. The absence of the UvrD protein stimulated the recombination between the direct repeats whereas the absence of the DdrB protein, previously shown to be involved in DNA double strand break repair through a single strand annealing (SSA pathway, strongly reduces the frequency of RecA- (and RecO- independent deletions events. The absence of the DdrB protein also increased the lethal sectoring of cells devoid of RecA or RecO protein. γ-irradiation of recA+ cells increased about 10-fold the frequencies of the deletion events, but at a lesser extend in cells devoid of the DdrB protein. Altogether, our results suggest a major role of single strand annealing in DNA repeat deletion events in bacteria devoid of the RecA protein, and also in recA+ bacteria exposed to ionizing radiation.

  11. Terapias Cognitiva e Cognitivo-Comportamental em dependência química

    OpenAIRE

    Silva,Cláudio Jerônimo da; Serra,Ana Maria

    2004-01-01

    Este artigo descreve o estado atual da Terapia Cognitiva, Comportamental, Prevenção de Recaída e Treinamento de Habilidades no tratamento de usuários de drogas. O objetivo é apresentar uma revisão sobre teorias e técnicas da Terapia Cognitiva e outras abordagens que dela derivam. Terapias Cognitiva e Comportamental, bem como Prevenção da Recaída e Treinamento de Habilidades, são tratamentos limitados no tempo, orientados em uma meta, e que utilizam sessões estruturadas, assumindo, assim, uma ...

  12. Current strategy for leprosy control in Brazil: time to pursue alternative preventive strategies?

    Directory of Open Access Journals (Sweden)

    Sérgio S. Cunha

    2004-11-01

    Full Text Available La estrategia actual para el control de la lepra en Brasil se basa en dos actividades principales: la detección precoz de casos y el tratamiento de casos con farmacoterapia combinada. Además de dichas medidas, se realizan esfuerzos complementarios para identificar los contactos domésticos para el diagnóstico precoz y la vacunación con el bacilo de Calmette-Guérin (BCG. Sin embargo, la eficacia de estas acciones a la hora de reducir la incidencia de la lepra es aún discutible. Esto genera dudas acerca de la factibilidad de eliminar la lepra en Brasil e indica que deberían adoptarse otras medidas de prevención. A pesar del hecho de que la vacunación de los contactos se practica desde hace varios años, falta información sobre el impacto de esta medida sobre la incidencia de la lepra en la comunidad. Además, con frecuencia no se tiene en cuenta el impacto de la vacunación neonatal con la vacuna BCG. Sostenemos que hay lugar para la investigación de métodos alternativos en el control y recomendamos que el programa brasileño de control haga un seguimiento esmerado de las tasas de cobertura para la vacunación neonatal en aquellas zonas donde la lepra es endémica, con el fin de lograr que la cobertura siga siendo elevada. Además, recomendamos la realización de estudios, orientados hacia el desarrollo de políticas, sobre la eficacia, factibilidad e impacto de estrategias alternativas más previsoras. Algunos posibles temas para estos estudios serían: 1 el impacto de la vacunación de contactos sobre la incidencia de la enfermedad en la población en general, 2 la factibilidad y el impacto de la quimioprofilaxis, y 3 el impacto de la identificación de grupos de alto riesgo (o mediante la adopción de una definición amplia de los contactos expuestos a la lepra, o mediante la creación de nuevas herramientas diagnósticas, y del diseño de programas para la detección precoz y la administración de farmacoterapia combinada espec

  13. Modification of human beta-globin locus PAC clones by homologous recombination in Escherichia coli

    NARCIS (Netherlands)

    G.P. Patrinos (George); M. de Krom (Mariken); S. Bottardi; R.J. Janssens; E. Katsantoni (Eleni); A.W. Wai; D.J. Sherratt; F.G. Grosveld (Frank); A.M.A. Imam (Ali)

    2000-01-01

    textabstractWe report here modifications of human beta-globin PAC clones by homologous recombination in Escherichia coli DH10B, utilising a plasmid temperature sensitive for replication, the recA gene and a wild-type copy of the rpsL gene which allows for an efficient selection for

  14. Dynamic protein assemblies in homologous recombination with single DNA molecules

    NARCIS (Netherlands)

    van der Heijden, A.H.

    2007-01-01

    What happens when your DNA breaks? This thesis describes experimental work on the single-molecule level focusing on the interaction between DNA and DNA-repair proteins, in particular bacterial RecA and human Rad51, involved in homologous recombination. Homologous recombination and its central event

  15. Regulation of Recombination between gtfB/gtfC Genes in Streptococcus mutans by Recombinase A

    Directory of Open Access Journals (Sweden)

    Satoko Inagaki

    2013-01-01

    Full Text Available Streptococcus mutans produces 3 types of glucosyltransferases (GTFs, whose cooperative action is essential for cellular adhesion. The recombinase A (RecA protein is required for homologous recombination. In our previous study, we isolated several strains with a smooth colony morphology and low GTF activity, characteristics speculated to be derived from the GTF fusions. The purpose of the present study was to investigate the mechanism of those fusions. S. mutans strain MT8148 was grown in the presence of recombinant RecA (rRecA protein, after which smooth colonies were isolated. The biological functions and sequences of the gtfB and gtfC genes of this as well as other clinical strains were determined. The sucrose-dependent adherence rates of those strains were reduced as compared to that of MT8148. Determination of the sequences of the gtfB and gtfC genes showed that an approximately 3500 bp region was deleted from the area between them. Furthermore, expression of the recA gene was elevated in those strains as compared to MT8148. These results suggest that RecA has an important role in fusions of gtfB and gtfC genes, leading to alteration of colony morphology and reduction in sucrose-dependent adhesion.

  16. Subtipos moleculares de PML/RARα en pacientes con leucemia promielocítica aguda

    Directory of Open Access Journals (Sweden)

    María del Carmen Castro-Mujica

    2013-03-01

    Full Text Available El objetivo fue describir la frecuencia de los subtipos moleculares de PML/RARα en pacientes con leucemia promielocítica aguda (LPA y su distribución según grupo de riesgo de recaída y citomorfología. Se realizó una serie de casos que incluyó a cincuenta pacientes registrados en el Instituto Nacional de Enfermedades Neoplásicas (INEN, durante el periodo 2010-2012, con diagnóstico molecular de LPA PML/RARα y subtipos bcr1, bcr2 y bcr3 por reacción en cadena de la polimerasa con transcriptasa reversa (RT-PCR. El subtipo bcr1 fue el más frecuente (62%. Los pacientes con riesgo de recaída intermedio y morfología hipergranular fueron, en su mayoría, bcr1 (70% y todos los que poseían riesgo de recaída alto y morfología hipogranular fueron bcr3. Se concluye que en la población estudiada hay un predomino del subtipo bcr1 y que existen diferencias en la distribución de los subtipos bcr1 y bcr3 según el grupo de riesgo de recaída y citomorfología

  17. Resonancia magnética de la columna lumbar intervenida por hernia discal

    OpenAIRE

    Naranjo Santana, Pedro Antonio

    1999-01-01

    [ES] La Resonancia Magnética de la Columna Lumbar previa, y tras la administración de Gadolinio, se propone como prueba diagnóstica de primera elección de los pacientes intervenidos por hernia discal que presentan recaída de la sintomatología.

  18. Vibrio galatheae sp. nov., a novel member of the Vibrionaceae family isolated from the Solomon Sea

    DEFF Research Database (Denmark)

    Giubergia, Sonia; Machado, Henrique; Mateiu, Ramona Valentina

    2015-01-01

    RNA and fur genes indicated the affiliation of the strain to a new species. This observation was supported by a multilocus sequence analysis (MLSA) including sequences of the housekeeping genes 16S rRNA, gyrB, pyrH, recA and topA. In silico DNA-DNA hybridization (DDH) and Average Nucleotide Identity (ANI...

  19. Whole genome sequencing as a tool for phylogenetic analysis of clinical strains of Mitis group streptococci

    DEFF Research Database (Denmark)

    Rasmusen, L. H.; Dargis, R.; Iversen, Katrine Højholt

    2016-01-01

    with infective endocarditis were whole genome sequenced. We compared the phylogenetic analyses based on single genes (recA, sodA, gdh), multigene (MLSA), SNPs, and core-genome sequences. The six phylogenetic analyses generally showed a similar pattern of six monophyletic clusters, though a few differences were...

  20. SOS Induction by Stabilized Topoisomerase IA Cleavage Complex Occurs via the RecBCD Pathway▿ †

    OpenAIRE

    Sutherland, Jeanette H.; Cheng, Bokun; Liu, I-Fen; Tse-Dinh, Yuk-Ching

    2008-01-01

    Accumulation of mutant topoisomerase I cleavage complex can lead to SOS induction and cell death in Escherichia coli. The single-stranded break associated with mutant topoisomerase I cleavage complex is converted to double-stranded break, which then is processed by the RecBCD pathway, followed by association of RecA with the single-stranded DNA.

  1. Mechanisms of Mutation in Non-Dividing Cells

    National Research Council Canada - National Science Library

    Petrosino, Joseph

    2002-01-01

    .... Previously, our laboratory discovered that RecA (an hRAD51 homolog) and RecBCD recombination repair proteins are necessary for the acquisition of 13-lactam drug-resistant mutations in the Escherichia coli chromosome during stationary-phase...

  2. Nutritional assessment in children with cystic fibrosis

    Science.gov (United States)

    Optimal nutrition, including consuming 35–40% of calories (kcal) as fat, is a vital part of the management of cystic fibrosis (CF), and involves accurate assessment of dietary intake. We compared 3 methods of nutritional assessment in 8– to 14-year-old children (n=20) with CF: 1) a 24-h Dietary Reca...

  3. Structures of thymidine kinase 1 of human and mycoplasma origin

    DEFF Research Database (Denmark)

    Welin, Martin; Kosinska, Urszula; Mikkelsen, Nils-Egil

    2004-01-01

    , in complex with the feedback inhibitor dTTP. The TK1s have a tetrameric structure where each subunit contains an alfa/beta-domain that is similar to ATPase domains of members of the RecA structural family and a domain of a new type containing a structural zinc. The zinc ion connects beta...

  4. Psychosocial and health impacts of uranium mining and milling on Navajo lands.

    Science.gov (United States)

    Dawson, Susan E; Madsen, Gary E

    2011-11-01

    The uranium industry in the American Southwest has had profoundly negative impacts on American Indian communities. Navajo workers experienced significant health problems, including lung cancer and nonmalignant respiratory diseases, and psychosocial problems, such as depression and anxiety. There were four uranium processing mills and approximately 1,200 uranium mines on the Navajo Nation's over 27,000 square miles. In this paper, a chronology is presented of how uranium mining and milling impacted the lives of Navajo workers and their families. Local community leaders organized meetings across the reservation to inform workers and their families about the relationship between worker exposures and possible health problems. A reservation-wide effort resulted in activists working with political leaders and attorneys to write radiation compensation legislation, which was passed in 1990 as the Radiation Exposure Compensation Act (RECA) and included underground uranium miners, atomic downwinders, and nuclear test-site workers. Later efforts resulted in the inclusion of surface miners, ore truck haulers, and millworkers in the RECA Amendments of 2000. On the Navajo Nation, the Office of Navajo Uranium Workers was created to assist workers and their families to apply for RECA funds. Present issues concerning the Navajo and other uranium-impacted groups include those who worked in mining and milling after 1971 and are excluded from RECA. Perceptions about uranium health impacts have contributed recently to the Navajo people rejecting a resumption of uranium mining and milling on Navajo lands.

  5. Novel recombinant ethyl ferulate esterase from Burkholderia multivorans

    CSIR Research Space (South Africa)

    Rashamuse, KJ

    2007-11-01

    Full Text Available techniques. Detailed molecular identification based on species-specific primers and two conserved genes (16S rRNA and recA) led to the identification of the isolate as Burkholderia multivorans UWC10. A gene (designated estEFH5) encoding an EFH enzyme...

  6. A dominant, recombination-defective allele of Dmc1 causing male-specific sterility

    NARCIS (Netherlands)

    Bannister, Laura A.; Pezza, Roberto J.; Donaldson, Janet R.; de Rooij, Dirk G.; Schimenti, Kerry J.; Camerini-Otero, R. Daniel; Schimenti, John C.

    2007-01-01

    DMC1 is a meiosis-specific homolog of bacterial RecA and eukaryotic RAD51 that can catalyze homologous DNA strand invasion and D-loop formation in vitro. DMC1-deficient mice and yeast are sterile due to defective meiotic recombination and chromosome synapsis. The authors identified a male dominant

  7. Induction of genetic recombination in the lambda bacteriophage by ultraviolet radiation of the Escherichia Coli cells

    International Nuclear Information System (INIS)

    Alcantara D, D.

    1986-12-01

    In this work there are reported the results that show that although the stimulation of the recombination of the Lambda bacteriophage, by UV irradiation of the cells of Escherichia Coli, it looks to be the result of the high expression of the functions of the SOS system, doesn't keep some relationship with the high concentration of protein reached RecA. (Author)

  8. Chromosomal Fragmentation in "Escherichia Coli": Its Absence in "mutT" Mutants and Its Mechanisms in "seqA" Mutants

    Science.gov (United States)

    Rotman, Ella Rose

    2009-01-01

    Chromosomal fragmentation in "Escherichia coli" is a lethal event for the cell unless mended by the recombinational repair proteins RecA, RecBCD, and RuvABC. Certain mutations exacerbate problems that cause the cell to be dependent on the recombinational repair proteins for viability. We tested whether the absence of the MutT protein caused…

  9. The RecA-Dependent SOS Response Is Active and Required for Processing of DNA Damage during Bacillus subtilis Sporulation.

    Directory of Open Access Journals (Sweden)

    Fernando H Ramírez-Guadiana

    Full Text Available The expression of and role played by RecA in protecting sporulating cells of Bacillus subtilis from DNA damage has been determined. Results showed that the DNA-alkylating agent Mitomycin-C (M-C activated expression of a PrecA-gfpmut3a fusion in both sporulating cells' mother cell and forespore compartments. The expression levels of a recA-lacZ fusion were significantly lower in sporulating than in growing cells. However, M-C induced levels of ß-galactosidase from a recA-lacZ fusion ~6- and 3-fold in the mother cell and forespore compartments of B. subtilis sporangia, respectively. Disruption of recA slowed sporulation and sensitized sporulating cells to M-C and UV-C radiation, and the M-C and UV-C sensitivity of sporangia lacking the transcriptional repair-coupling factor Mfd was significantly increased by loss of RecA. We postulate that when DNA damage is encountered during sporulation, RecA activates the SOS response thus providing sporangia with the repair machinery to process DNA lesions that may compromise the spatio-temporal expression of genes that are essential for efficient spore formation.

  10. Genetic diversity of Iranian potato soft rot bacteria based on PCR ...

    African Journals Online (AJOL)

    TENEBRA

    2012-01-19

    Jan 19, 2012 ... Key words: Genetic diversity, polymerase chain reaction-restriction fragment length polymorphism (PCR-. RFLP), recA gene, Pectobacterium spp. .... each restriction enzyme was used for digestions, which were incubated overnight at the .... (2006) with application of the molecular marker of PCR-RFLP, ...

  11. Análisis de costo-efectividad en Colombia de anastrazol Vs. tamoxifeno como terapia inicial en mujeres con cáncer temprano de mama y receptor hormonal positivo

    Directory of Open Access Journals (Sweden)

    Óscar Gamboa

    2010-03-01

    Conclusiones. La utilización de anastrazol genera un costo por año libre de recaída superior al producto interno bruto per cápita de Colombia (Col$ 7’521.363 para 2007. En consecuencia, resulta recomendable para Colombia continuar con el tamoxifeno por cinco años.

  12. Influencia del nivel de conocimiento sobre sexualidad en los tipos de actitud respecto a los contenidos de sexualidad de los docentes del área de Persona, Familia y Relaciones Humanas del nivel secundario pertenecientes en la UGEL 01 de Lima Metropolitana

    OpenAIRE

    Rivera Bravo, Luisa Vivian

    2015-01-01

    Identifica la influencia del nivel de conocimiento de los contenidos de sexualidad en los tipos de actitud de los docentes del área de Persona, Familia y relaciones Humanas del nivel secundario, la cual recae en las siguientes dimensiones: las características sociales en la adolescencia, los valores, creencias y prejuicios sobre sexualidad y las relaciones interpersonales. Tesis

  13. Genetic diversity of Iranian potato soft rot bacteria based on ...

    African Journals Online (AJOL)

    Based on biochemical and physiological tests, the Iranian strains were identified as either Pectobacterium carotovorum subsp. carotovorum or P. carotovorum subsp. wasabiae. Sequence analysis of recA gene revealed that the strains closely related to Pectobacterium spp. To assess the genomic diversity, polymerase ...

  14. Ensifer, Phyllobacterium and Rhizobium species occupy nodules of Medicago sativa (alfalfa) and Melilotus alba (sweet clover) grown at a Canadian site without a history of cultivation

    Science.gov (United States)

    Phage resistant and susceptible bacteria from nodules of alfalfa and sweet clover grown at a site without a known history of cultivation, were identified as Ensifer, Rhizobium and Phyllobacterium species based on sequence analysis of ribosomal (16S and 23S rRNA)and protein encoding (atpD and recA) g...

  15. Different efficiency of UmuDC and MucAB proteins in UV light induced mutagenesis in Escherichia coli

    International Nuclear Information System (INIS)

    Blanco, M.; Herrera, G.; Aleixandre, V.

    1986-01-01

    Two multicopy plasmids carrying either the umuDC or the mucAB operon were used to compare the efficiency of UmuDC and MucAB proteins in UV mutagenesis of Escherichia coli K12. It was found that in recA + uvr + bacteria, plasmid pIC80, mucAB + mediated UV mutagenesis more efficiently than did plasmid pSE 117, umuDC + . A similar result was obtained in lex A51(Def) cells, excluding the possibility that this was due to a differential regulation by LexA of the umuDC and mucAB operons. We conclude that some structural characteristic of the UmuDC and MucAB proteins determines their different efficiency in UV mutagenesis. This characteristic could be also responsible for the observation that in the recA430 mutant, pIC80 but no pSE117 can mediate UV mutagenesis. In the recAS142 mutant pIC80 also promoted UV mutagenesis more efficiently than pSE117. In this mutant, the recombination proficiency, the protease activity toward LexA and the mutation frequency were increased by the presence of adenine in the medium. In recA + uvrB5 bacteria, plasmid pSE117, umuDC caused both an increase in UV sensitivity as well as a reduction in the mutation frequency. These negative effects resulting from the overproduction of UmuDC proteins were higher in recA142 uvrB5 than in recA + uvrB5 cells. In contrast, overproduction of MucAB proteins in excision-deficient bacteria containing pIC80 led to a large increase in the mutation frequency. We suggest that the functional differences between UmuDC and MucAB proteins might be due to their different dependence on the direct role of RecA protease in UV mutagenesis. (orig.)

  16. Segmental isotope labeling of proteins for NMR structural study using a protein S tag for higher expression and solubility

    International Nuclear Information System (INIS)

    Kobayashi, Hiroshi; Swapna, G. V. T.; Wu, Kuen-Phon; Afinogenova, Yuliya; Conover, Kenith; Mao, Binchen; Montelione, Gaetano T.; Inouye, Masayori

    2012-01-01

    A common obstacle to NMR studies of proteins is sample preparation. In many cases, proteins targeted for NMR studies are poorly expressed and/or expressed in insoluble forms. Here, we describe a novel approach to overcome these problems. In the protein S tag-intein (PSTI) technology, two tandem 92-residue N-terminal domains of protein S (PrS 2 ) from Myxococcus xanthus is fused at the N-terminal end of a protein to enhance its expression and solubility. Using intein technology, the isotope-labeled PrS 2 -tag is replaced with non-isotope labeled PrS 2 -tag, silencing the NMR signals from PrS 2 -tag in isotope-filtered 1 H-detected NMR experiments. This method was applied to the E. coli ribosome binding factor A (RbfA), which aggregates and precipitates in the absence of a solubilization tag unless the C-terminal 25-residue segment is deleted (RbfAΔ25). Using the PrS 2 -tag, full-length well-behaved RbfA samples could be successfully prepared for NMR studies. PrS 2 (non-labeled)-tagged RbfA (isotope-labeled) was produced with the use of the intein approach. The well-resolved TROSY-HSQC spectrum of full-length PrS 2 -tagged RbfA superimposes with the TROSY-HSQC spectrum of RbfAΔ25, indicating that PrS 2 -tag does not affect the structure of the protein to which it is fused. Using a smaller PrS-tag, consisting of a single N-terminal domain of protein S, triple resonance experiments were performed, and most of the backbone 1 H, 15 N and 13 C resonance assignments for full-length E. coli RbfA were determined. Analysis of these chemical shift data with the Chemical Shift Index and heteronuclear 1 H– 15 N NOE measurements reveal the dynamic nature of the C-terminal segment of the full-length RbfA protein, which could not be inferred using the truncated RbfAΔ25 construct. CS-Rosetta calculations also demonstrate that the core structure of full-length RbfA is similar to that of the RbfAΔ25 construct.

  17. Evaluation of polymerase chain reaction in nerve biopsy specimens of patients with Hansen's disease.

    Science.gov (United States)

    Tiwari, Vandana; Malhotra, Kiranpreet; Khan, Kainat; Maurya, Pradeep K; Singh, Ajai Kumar; Thacker, Anup Kumar; Husain, Nuzhat; Kulshreshtha, Dinkar

    2017-09-15

    Pure neuritic variety of leprosy (PNL) presents as peripheral neuropathy with absent skin lesions and negative skin smears. Diagnosing PNL is an uphill task as most of these patients have nonspecific changes on nerve biopsy. In such circumstances, additional molecular diagnostic tools like polymerase chain reaction (PCR) has proven to be useful in diagnosing leprosy. The present study was planned to evaluate the role of PCR in nerve biopsy specimens of patients with PNL. Patients attending the neuromuscular clinic from January 2013 to June 2014 with mononeuropathy multiplex underwent detailed diagnostic evaluation to ascertain the cause of neuropathy. Patients where this evaluation failed to establish an etiology underwent a nerve biopsy. Nerve biopsy was done in 52 patients, of which 35 were diagnosed as pure neuritic leprosy. Definite leprosy with positive wade fite staining for lepra bacilli was seen in 13 patients and 22 biopsies revealed a probable leprosy without lepra bacilli being identified. PCR for M. leprae was positive in 22 patients (62%). 12 of the 13 cases with definite leprosy on histopathology were PCR positive while in the AFB negative group, PCR was positive in 10 cases. PCR had a sensitivity of 92.3%, specificity of 54.5%. The positive and negative predictive value of PCR was 54.5% and 92.3% respectively. PCR helps in diagnosing PNL in doubtful cases. A positive PCR increases the sensitivity of detection of M. leprae especially in cases of probable PNL group where AFB cannot be demonstrated on histopathology. Copyright © 2017 Elsevier B.V. All rights reserved.

  18. Use of skin graft "punch graft" type for the healing of leg ulcers in treated hansen's disease patients

    OpenAIRE

    Carla Christiane de Oliveira Cardia

    2006-01-01

    Hansen's disease is an infectious illness caused by Mycobacterium leprae. It affects preferentially the skin and the peripheral nervous system leading to incapacities, such as leg ulcers, which happens due to the direct action of the bacillus on the organs or its indirect action on the peripheral nervous system. Leg ulcers can occur by two physiopathologic processes. There are many treatments for general leg ulcers, which include the ones caused by Hansen's disease sequels. Among them, surgic...

  19. Hansen's disease mimicking a systemic vasculitis.

    Science.gov (United States)

    Sampaio, L; Silva, L; Terroso, G; Pimenta, S; Brandão, F; Pinto, J; Prisca, A; Brito, J; Ventura, F

    2011-01-01

    Hansen's disease, caused by Mycobacterium leprae, classically presents with cutaneous and neurological manifestations. Rheumatologic manifestations present in 1 to 5% of the patients, and include arthritis, arthralgias, Charcot arthropathy, erythema nodosum and vasculitis. We report a case of a 86 year old woman with polyarthritis, subcutaneous nodules and leg ulcers whose differential diagnosis included primary vasculitis and diffuse connective tissue diseases and ended to be leprosy in a non endemic country.

  20. A possible dysfunction of melanosome transfer in leprosy: an electron-microscopic study.

    Science.gov (United States)

    Westerhof, W

    1977-01-01

    An E.M. study was carried out to investigate whether Mycobacterium leprae occur intracellularly in epidermal melanocytes. As this could not be confirmed, the selective killing of melanocytes by cytotoxic lymphocytes could not explain the hypopigmentation in types of leprosy with a relative good immune response. There were indications that these hypopigmented lesions resulted from a disturbed transfer of melanosomes from melanocytes to keratinocytes. Further research is in progress.

  1. Investigating potential exogenous tumor initiating and promoting factors for Cutaneous T-Cell Lymphomas (CTCL), a rare skin malignancy

    DEFF Research Database (Denmark)

    Litvinov, Ivan V.; Shtreis, Anna; Kobayashi, Kenneth

    2016-01-01

    Most skin malignancies are caused by external and often preventable environmental agents. Multiple reports demonstrated that cutaneous T-cell lymphomas (CTCL) can occur in married couples and cluster in families. Furthermore, recent studies document geographic clustering of this malignancy in Texas...... as well as in other areas of the United States. Multiple infectious, occupational, and medication causes have been proposed as triggers or promoters of this malignancy including hydrochlorothiazide diuretics, Staphylococcus aureus, dermatophytes, Mycobacterium leprae, Chlamydia pneumoniae, human T...

  2. Clinic-epidemiological evaluation of ulcers in patients with leprosy sequelae and the effect of low level laser therapy on wound healing: a randomized clinical trial

    OpenAIRE

    Barreto, Josaf? G; Salgado, Claudio G

    2010-01-01

    Abstract Background Mycobacterium leprae is the only pathogenic bacteria able to infect peripheral nerves. Neural impairment results in a set of sensitive, motor and autonomic disturbances, with ulcers originating primarily on the hands and feet. The study objectives were to analyze the clinic-epidemiological characteristics of patients attended at one specialized dressing service from a leprosy-endemic region of the Brazilian Amazon and to evaluate the effect of low level laser therapy (LLLT...

  3. Whole genome sequencing distinguishes between relapse and reinfection in recurrent leprosy cases.

    Directory of Open Access Journals (Sweden)

    Mariane M A Stefani

    2017-06-01

    Full Text Available Since leprosy is both treated and controlled by multidrug therapy (MDT it is important to monitor recurrent cases for drug resistance and to distinguish between relapse and reinfection as a means of assessing therapeutic efficacy. All three objectives can be reached with single nucleotide resolution using next generation sequencing and bioinformatics analysis of Mycobacterium leprae DNA present in human skin.DNA was isolated by means of optimized extraction and enrichment methods from samples from three recurrent cases in leprosy patients participating in an open-label, randomized, controlled clinical trial of uniform MDT in Brazil (U-MDT/CT-BR. Genome-wide sequencing of M. leprae was performed and the resultant sequence assemblies analyzed in silico.In all three cases, no mutations responsible for resistance to rifampicin, dapsone and ofloxacin were found, thus eliminating drug resistance as a possible cause of disease recurrence. However, sequence differences were detected between the strains from the first and second disease episodes in all three patients. In one case, clear evidence was obtained for reinfection with an unrelated strain whereas in the other two cases, relapse appeared more probable.This is the first report of using M. leprae whole genome sequencing to reveal that treated and cured leprosy patients who remain in endemic areas can be reinfected by another strain. Next generation sequencing can be applied reliably to M. leprae DNA extracted from biopsies to discriminate between cases of relapse and reinfection, thereby providing a powerful tool for evaluating different outcomes of therapeutic regimens and for following disease transmission.

  4. An unusual case of isolated sixth cranial nerve palsy in leprosy.

    Science.gov (United States)

    Vaishampayan, Sanjeev; Borde, Priyanka

    2012-08-15

    Cranial nerve involvement is not common in leprosy. The fifth and seventh cranial nerves are the most commonly affected in leprosy. Herein we present a patient with Hansen disease (BL) with type I reaction who developed isolated involvement of the sixth cranial nerve leading to lateral rectus muscle palsy. He responded to timely anti-reactional therapy and it produced a good response. Careful observation of patients with lepra reaction is needed to avoid damage to important organs.

  5. Recent advances in leprosy and Buruli ulcer (Mycobacterium ulcerans infection).

    Science.gov (United States)

    Walsh, Douglas S; Portaels, Françoise; Meyers, Wayne M

    2010-10-01

    After tuberculosis, leprosy (Mycobacterium leprae) and Buruli ulcer (M. ulcerans infection) are the second and third most common mycobacterial infections in humankind, respectively. Recent advances in both diseases are summarized. Leprosy remains a public health problem in some countries, and new case detections indicate active transmission. Newly identified M. lepromatosis, closely related to M. leprae, may cause disseminated leprosy in some regions. In genome-wide screening in China, leprosy susceptibility associates with polymorphisms in seven genes, many involved with innate immunity. World Health Organization multiple drug therapy administered for 1 or 2 years effectively arrests disseminated leprosy but disability remains a public health concern. Relapse is infrequent, often associated with higher pretreatment M. leprae burdens. M. ulcerans, a re-emerging environmental organism, arose from M. marinum and acquired a virulence plasmid coding for mycolactone, a necrotizing, immunosuppressive toxin. Geographically, there are multiple strains of M. ulcerans, with variable pathogenicity and immunogenicity. Molecular epidemiology is describing M. ulcerans evolution and genotypic variants. First-line therapy for Buruli ulcer is rifampin + streptomycin, sometimes with surgery, but improved regimens are needed. Leprosy and Buruli ulcer are important infections with significant public health implications. Modern research is providing new insights into molecular epidemiology and pathogenesis, boding well for improved control strategies.

  6. Editoriales

    Directory of Open Access Journals (Sweden)

    Facultad de Medicina Revista

    1941-07-01

    Full Text Available Décimo año de vida Bajo muy alagadores auspicios, inicia hoy su décimo año de vida el órgano de la Facultad de Medicina. El prestigio de la Revista está asegurado en los centros científicos universales. Sus escritos son citados y comentados con elogio y las publicaciones de todos los países, se manifiestan muy honradas en establecer canje y mantener relaciones permanentes con nuestra Revista. La profilaxis de la lepra Son postulados científicos inmutables, que la lepra se contrae principalmente en la niñez, cuando el niño vive permanentemente en ambiente contaminado y que si el hijo de enfermos se separa temprano de la fuente de contagio, se libra de la lepra. El Departamento Nacional de Higiene Repetidamente hemos oído hablar de que las Cámaras Legislativas, reformarán otra vez las directivas de la salubridad pública. Aún cuando nadie nos ha preguntado nuestro parecer, queremos expresar nuestro pensamiento al respecto.

  7. Topoisomerase I function during Escherichia coli response to antibiotics and stress enhances cell killing from stabilization of its cleavage complex

    Science.gov (United States)

    Liu, I-Fen; Sutherland, Jeanette H.; Cheng, Bokun; Tse-Dinh, Yuk-Ching

    2011-01-01

    Objectives To explore the role of topoisomerase I in gene activation and increased RecA levels during the bacterial SOS response, as well as the effect of antibiotic treatment and stress challenge on cell killing initiated by trapped topoisomerase I cleavage complex. Methods A mutant Escherichia coli strain with a ΔtopA mutation was used to investigate the role of topoisomerase I function in the SOS response to trimethoprim and mitomycin C. Induction of the recA and dinD1 promoters was measured using luciferase reporters of these promoters fused to luxCDABE. An increase in the RecA level following trimethoprim treatment was quantified directly by western blotting. The effect of stress challenge from trimethoprim and acidified nitrite treatments on cell killing by topoisomerase I cleavage complex accumulation was measured by the decrease in viability following induction of recombinant mutant topoisomerase I that forms a stabilized cleavage complex. Results Topoisomerase I function was found to be required for efficient transcriptional activation of the recA and dinD1 promoters during the E. coli SOS response to trimethoprim and mitomycin C. The role of topoisomerase I in the SOS response was confirmed with quantitative western blot analysis of RecA following trimethoprim treatment. The bactericidal effect from topoisomerase I cleavage complex accumulation was shown to be enhanced by stress challenge from trimethoprim and acidified nitrite. Conclusions Bacterial topoisomerase I function is actively involved in the SOS response to antibiotics and stress challenge. Cell killing initiated by the topoisomerase I cleavage complex would be enhanced by antibiotics and the host response. These findings provide further support for bacterial topoisomerase I as a therapeutic target. PMID:21486853

  8. Recombinant protein hydrazides: application to site-specific protein PEGylation.

    Science.gov (United States)

    Thom, Jennifer; Anderson, David; McGregor, Joanne; Cotton, Graham

    2011-06-15

    Here, we describe a novel method for the site-specific C-terminal PEGylation of recombinant proteins. This general approach exploits chemical cleavage of precursor intein-fusion proteins with hydrazine to directly produce recombinant protein hydrazides. This unique functionality within the protein sequence then facilitates site-specific C-terminal modification by hydrazone-forming ligation reactions. This approach was used to generate folded, site-specifically C-terminal PEGylated IFNalpha2b and IFNbeta1b, which retained excellent antiviral activity, demonstrating the utility of this technology in the PEGylation of therapeutic proteins. As this methodology is straightforward to perform, is compatible with disulfide bonds, and is exclusively selective for the protein C-terminus, it shows great potential as general technology for the site-specific engineering and labeling of recombinant proteins.

  9. CHI 2013 Human Work Interaction Design (HWID) SIG

    DEFF Research Database (Denmark)

    Clemmensen, Torkil; Campos, Pedro F.; Katre, Dinesh S.

    2013-01-01

    In this SIG we aim to introduce the IFIP 13.6 Human Work Interaction Design (HWID) approach to the CHI audience. The HWID working group aims at establishing relationships between extensive empirical work-domain studies and HCI design. We invite participants from industry and academia with an inte......In this SIG we aim to introduce the IFIP 13.6 Human Work Interaction Design (HWID) approach to the CHI audience. The HWID working group aims at establishing relationships between extensive empirical work-domain studies and HCI design. We invite participants from industry and academia...... with an interest on empirical work analysis, HCI, interaction design and usability and user experience in work situations and in the workplace. This SIG is a vital step towards creating a CHI2014 workshop on this topic....

  10. Characterization of mitochondrial thioredoxin reductase from C. elegans

    International Nuclear Information System (INIS)

    Lacey, Brian M.; Hondal, Robert J.

    2006-01-01

    Thioredoxin reductase catalyzes the NADPH-dependent reduction of the catalytic disulfide bond of thioredoxin. In mammals and other higher eukaryotes, thioredoxin reductases contain the rare amino acid selenocysteine at the active site. The mitochondrial enzyme from Caenorhabditis elegans, however, contains a cysteine residue in place of selenocysteine. The mitochondrial C. elegans thioredoxin reductase was cloned from an expressed sequence tag and then produced in Escherichia coli as an intein-fusion protein. The purified recombinant enzyme has a k cat of 610 min -1 and a K m of 610 μM using E. coli thioredoxin as substrate. The reported k cat is 25% of the k cat of the mammalian enzyme and is 43-fold higher than a cysteine mutant of mammalian thioredoxin reductase. The enzyme would reduce selenocysteine, but not hydrogen peroxide or insulin. The flanking glycine residues of the GCCG motif were mutated to serine. The mutants improved substrate binding, but decreased the catalytic rate

  11. Split-gene system for hybrid wheat seed production.

    Science.gov (United States)

    Kempe, Katja; Rubtsova, Myroslava; Gils, Mario

    2014-06-24

    Hybrid wheat plants are superior in yield and growth characteristics compared with their homozygous parents. The commercial production of wheat hybrids is difficult because of the inbreeding nature of wheat and the lack of a practical fertility control that enforces outcrossing. We describe a hybrid wheat system that relies on the expression of a phytotoxic barnase and provides for male sterility. The barnase coding information is divided and distributed at two loci that are located on allelic positions of the host chromosome and are therefore "linked in repulsion." Functional complementation of the loci is achieved through coexpression of the barnase fragments and intein-mediated ligation of the barnase protein fragments. This system allows for growth and maintenance of male-sterile female crossing partners, whereas the hybrids are fertile. The technology does not require fertility restorers and is based solely on the genetic modification of the female crossing partner.

  12. Cyclic peptide inhibitors of the β-sliding clamp in Staphylococcus aureus

    DEFF Research Database (Denmark)

    Kjelstrup, Susanne; Hansen, Paula Melo Paulon; Thomsen, Line Elnif

    2013-01-01

    Interaction between pairs of Staphylococcus aureus replication proteins was detected in an Escherichia coli based two-hybrid analysis. A reverse two-hybrid system was constructed for selection of compounds that hindered interaction between interacting protein pairs. A number of cyclic peptides......, from a library generated by the split intein-mediated circular ligation of peptides and proteins technology, were found to interfere with dimerization of the β-sliding clamp of the replisome. Two 8-mer peptides were analyzed in more detail. Both inhibited DNA replication, led to SOS induction, altered...... cell morphology and cell death. The peptides were active when added to bacterial cultures indicating that they could traverse the bacterial membrane to find their intracellular target. Peptide specificity was confirmed by overproduction of the putative target (DnaN) which resulted in resistance...

  13. BCG and Adverse Events in the Context of Leprosy

    Directory of Open Access Journals (Sweden)

    Renate Richardus

    2018-04-01

    Full Text Available BackgroundNotwithstanding its beneficial immunoprophylactic outcomes regarding leprosy and childhood TB, BCG vaccination may cause adverse events, particularly of the skin. However, this local hyper-immune reactivity cannot be predicted before vaccination, nor is its association with protection against leprosy known. In this study we investigated the occurrence of adverse events after BCG (revaccination in contacts of leprosy patients and analyzed whether the concomitant systemic anti-mycobacterial immunity was associated with these skin manifestations.MethodsWithin a randomized controlled BCG vaccination trial in Bangladesh, 14,828 contacts of newly diagnosed leprosy patients received BCG vaccination between 2012 and 2017 and were examined for adverse events 8 to 12 weeks post-vaccination. From a selection of vaccinated contacts, venous blood was obtained at follow-up examination and stimulated with Mycobacterium leprae (M. leprae antigens in overnight whole-blood assays (WBA. M. leprae phenolic glycolipid-I-specific antibodies and 32 cytokines were determined in WBAs of 13 individuals with and 13 individuals without adverse events after vaccination.ResultsOut of the 14,828 contacts who received BCG vaccination, 50 (0.34% presented with adverse events, mainly (80% consisting of skin ulcers. Based on the presence of BCG scars, 30 of these contacts (60% had received BCG in this study as a booster vaccination. Similar to the pathological T-cell immunity observed for tuberculoid leprosy patients, contacts with adverse events at the site of BCG vaccination showed elevated IFN-γ levels in response to M. leprae-specific proteins in WBA. However, decreased levels of sCD40L in serum and GRO (CXCL1 in response to M. leprae simultaneously indicated less T-cell regulation in these individuals, potentially causing uncontrolled T-cell immunity damaging the skin.ConclusionSkin complications after BCG vaccination present surrogate markers for protective

  14. Design, preparation and use of ligated phosphoproteins: a novel approach to study protein phosphatases by dot blot array, ELISA and Western blot assays.

    Science.gov (United States)

    Sun, Luo; Ghosh, Inca; Barshevsky, Tanya; Kochinyan, Samvel; Xu, Ming-Qun

    2007-07-01

    The study of substrate specificity of protein phosphatases (PPs) is very challenging since it is difficult to prepare a suitable phosphorylated substrate. Phosphoproteins, phosphorylated by a protein kinase, or chemically synthesized phosphopeptides are commonly used substrates for PPs. Both types of these substrates have their advantages and limitations. Phosphoproteins mimic more closely the physiologically relevant PP substrates, but their preparation is technically demanding. Synthetic phosphopeptides present advantages over proteins because they can be easily produced in large quantity and their amino acid sequence can be designed to contain potential determinants of substrate specificity. However, short peptides are less optimal compared to in vivo PP substrates and often display poor and variable binding to different matrices, resulting in low sensitivity in analysis of PP activity on solid support. In this work we utilize the intein-mediated protein ligation (IPL) technique to generate substrates for PPs, combining the advantages of proteins and synthetic peptides in one molecule. The ligation of a synthetic phosphopeptide to an intein-generated carrier protein (CP) with a one-to-one stoichiometry results in the formation of a ligated phosphoprotein (LPP). Three widely used assays, dot blot array, Western blot and ELISA were employed to study the PP activity on LPP substrates. Dephosphorylation was measured by detection of the remaining phosphorylation, or lack of it, with a phospho-specific antibody. The data show the advantage of LPPs over free peptides in assays on solid supports. LPPs exhibited enhanced binding to the matrices used in the study, which significantly improved sensitivity and consistency of the assays. In addition, saturation of the signal was circumvented by serial dilution of the assay samples. This report describes detailed experimental procedures for preparation of LPP substrates and their use in PP assays based on immobilization on

  15. Characterization of the rec-1 gene of Haemophilus influenzae and behavior of the gene in Escherichia coli

    International Nuclear Information System (INIS)

    Setlow, J.K.; Spikes, D.; Griffin, K.

    1988-01-01

    The rec-1 gene of Haemophilus influenzae was cloned into a shuttle vector that replicates in Escherichia coli as well as in H. influenzae. The plasmid, called pRec1, complemented the defects of a rec-1 mutant in repair of UV damage, transformation, and ability of prophage to be induced by UV radiation. Although UV resistance and recombination were caused by pRec1 in E. coli recA mutants, UV induction of lambda and UV mutagenesis were not. We suggest that the ability of the H. influenzae Rec-1 protein to cause cleavage of repressors but not the recombinase function differs from that of the E. coli RecA protein

  16. Regulation of the E. coli SOS response by the lexA gene product

    International Nuclear Information System (INIS)

    Brent, R.

    1983-01-01

    In an Escherichia coli that is growing normally, transcription of many genes is repressed by the product of the lexA gene. If cellular DNA is damaged, proteolytically competent recA protein (recA protease) inactivates lexA protein and these genes are induced. Many of the cellular phenomena observed during the cellular response to DNA damage (the SOS response) are the consequence of the expression of these lexA-prepressed genes. Since the SOS response of E. coli has recently been the subject of a comprehensive review, in this paper I would like to concentrate on some modifications to the picture based on new data. 12 references, 2 figures

  17. REFLEXIONES EN TORNO A LA RESPONSABILIDAD CIVIL EN EL DERECHO AERONÁUTICO Y DE LOS PILOTOS

    Directory of Open Access Journals (Sweden)

    Dylan Vallejo Garcés

    2012-01-01

    Full Text Available El presente artículo pretende mostrar la responsabilidad que surge del dere­cho aeronáutico y la responsabilidad que recae sobre los pilotos en el mo­mento del transporte aéreo, cuál es el tipo de responsabilidad que en ambos casos puede endilgarse, además de conocer cuál es la autoridad competente para dirimir los conflictos que se presenten en torno a estos temas, bien sea en analizar los hechos generadores del daño, el daño causado, el nexo de causalidad que existe entre el hecho antijurídico y el daño producido y cuál es el grado de imputación sobre el cual recae la responsabilidad.

  18. Las obras testimoniales de Max Aub sobre la guerra de España: las difíciles memorias de la derrota y del exilio

    Directory of Open Access Journals (Sweden)

    Gérard Malgat

    2011-10-01

    Full Text Available ¿Sobre quién ha recaído y recae aún la responsabilidad de edificar la memoria pública de la guerra de España y del largo exilio de los republicanos españoles en 1939 ? ¿Sobre el historiador ? ¿El periodista ? ¿El poeta ? ¿El novelista ? ¿El dramaturgo ? ¿El cineasta ? ¿O el sobreviviente ? Dado que intervino en cada uno de estos campos, Max Aub fue y continúa siendo un portavoz destacado de la memoria republicana española.No se trata aquí de hacer una apología de este escritor o la exégesis d...

  19. Characterization of the rec-1 gene of Haemophilus influenzae and behavior of the gene in Escherichia coli

    Energy Technology Data Exchange (ETDEWEB)

    Setlow, J.K.; Spikes, D.; Griffin, K.

    1988-09-01

    The rec-1 gene of Haemophilus influenzae was cloned into a shuttle vector that replicates in Escherichia coli as well as in H. influenzae. The plasmid, called pRec1, complemented the defects of a rec-1 mutant in repair of UV damage, transformation, and ability of prophage to be induced by UV radiation. Although UV resistance and recombination were caused by pRec1 in E. coli recA mutants, UV induction of lambda and UV mutagenesis were not. We suggest that the ability of the H. influenzae Rec-1 protein to cause cleavage of repressors but not the recombinase function differs from that of the E. coli RecA protein.

  20. Ocorrência de Herminodes sp. (Lepidoptera: Noctuidae em pupunheira nos estados do Acre e Rondônia, Brasil Occurrence of Herminodes sp. (Lepidoptera: Noctuidae in peach palm in the States of Acre and Rondônia, Brazil

    Directory of Open Access Journals (Sweden)

    Marcílio José Thomazini

    2004-09-01

    Full Text Available É relatado a ocorrência de Herminodes sp. em plantas de pupunha na área experimental da Embrapa Acre, Rio Branco, AC e no Projeto Reca, Vila Nova Califórnia, RO, Brasil. São descritas as principais características deste inseto e os danos por ele causados. Este é o primeiro registro de lagartas do gênero Herminodes causando dano em pupunheiras nos estados do Acre e Rondônia.Related herein is the occurrence of Herminodes sp. in peach palm plants at experimental area of Embrapa Acre, Rio Branco, AC and at Projeto Reca, Vila Nova California, RO, Brazil. The main characteristics of this insect and the damage it causes are described. This is the first record of Herminodes caterpillars damaging peach palm in the states of Acre and Rondônia.

  1. Inducible pathway is required for mutagenesis in Salmonella typhimurium LT2

    International Nuclear Information System (INIS)

    Orrego, C.; Eisenstadt, E.

    1987-01-01

    UV mutability of Salmonella typhimurium LT2 was eliminated in the presence of a multicopy plasmid carrying the Escherichia coli lexA + gene. This result suggests that inducible, SOS-like functions are required for UV mutagenesis in S. typhimurium. S. typhimurium strains carrying either point or deletion mutations in topA had previously been shown to lose their mutability by UV or methyl methanesulfonate. Mitomycin C induction of the Phi(mucB'-lacZ') fusion (a DNA damage-inducible locus carried on plasmid pSE205) in S. typhimurium topA was normal, suggesting that RecA is activated in topA mutants. These observations lead the authors deduce that S. typhimurium has at least one DNA damage-inducible locus in addition to recA that is required for UV mutability

  2. Molecular cloning of a recA-like gene from the cyanobacterium Anabaena variabilis

    International Nuclear Information System (INIS)

    Owttrim, G.W.; Coleman, J.R.

    1987-01-01

    A recA-like gene isolated from the cyanobacterium Anabaena variabilis was cloned and partially characterized. When introduced into Escherichia coli recA mutants, the 7.5-kilobase-pair plasmid-borne DNA insert restored resistance to methyl methanesulfonate and UV irradiation, as well as recombination proficiency when measured by Hfr-mediated conjugation. The cyanobacterial recA gene restored spontaneous but not mitomycin C-induced prophage production. Restriction analysis and subcloning yielded a 1.5-kilobase-pair Sau3A fragment which also restored methylmethane sulfonate resistance and coded for a 38- to 40-kilodalton polypeptide when expressed in an in vitro transcription-translation system

  3. Importancia de la depresión en atención primaria : aspectos sociodemográficos

    OpenAIRE

    Aragonès Benaiges, Enric

    2006-01-01

    En población europea, la prevalencia de depresión mayor es del 3,9 % y la prevalencia de distimia es del 1,1 %. La duración media de los episodios de depresión mayor es de 27 semanas y una cuarta parte se cronifican. Las recaídas son frecuentes. El número de episodios previos y la persistencia de síntomas residuales son predictores de nuevas recaídas. En atención primaria la prevalencia de depresión es más alta que en población general: 14,3 % para la depresión mayor y 4,8 % para la distimia....

  4. La calidad del empleo para los hombres y las mujeres: un estudio desde la informalidad, el subempleo y las percepciones

    Directory of Open Access Journals (Sweden)

    Lina María Restrepo Plaza

    2013-01-01

    Full Text Available La tasa de desempleo, como un indicador de desempeño económico y social para países en desarrollo, suele arrojar falacias que no permiten dar cuenta de la manera como se ocupan los trabajadores de una sociedad. De este modo, surge la necesidad de estudiar qué está sucediendo con la calidad del empleo que, para efectos de este documento, se ha decidido abordar desde la informalidad, el subempleo y la satisfacción reportada por los trabajadores es sus ocupaciones. A partir de una descripción de algunas variables que forman parte de estas tres dimensiones, se logró determinar que, aunque hombres y mujeres manifiestan similares índices de satisfacción en sus trabajos, la informalidad recae en mayor medida en los hombres, mientras que el subempleo recae de forma similar en ambos sexos.

  5. Piomiositis tuberculosa en un paciente inmunosuprimido

    Directory of Open Access Journals (Sweden)

    Johanna Osorio

    2016-04-01

    El caso que se presenta es llamativo dada la rareza de esta manifestación de la tuberculosis y la lenta mejoría con el tratamiento antituberculoso de primera línea en este paciente con infección por HIV y recaída, a pesar de que el aislamiento microbiológico resultó sensible.

  6. Ultraviolet light protection, enhancement of ultraviolet light mutagenesis, and mutator effect of plasmid R46 in Salmonella typhimurium

    International Nuclear Information System (INIS)

    Mortelmans, K.E.; Stocker, B.A.D.

    1976-01-01

    Plasmid R46 partially protected Salmonella typhimurium, wild type or uvrB or polA, against the lethal effect of ultraviolet (uv) irradiation, but did not protect recA mutants. The plasmid also increased frequency of uv-induced reversion to His + in all tested his point mutants (wild type for uv sensitivity), including amber, ochre, UGA, missense, and frame-shift mutants. Plasmid R46 also increased uv-induced reversion to His + in uvrB and polA strains, but no uv mutagenic effect was detected in R - or R46-carrying recA derivatives of a his(amber) mutant. The spontaneous reversion frequency of his nonsense mutants of all classes, and of some his missense mutants, was increased about 10-fold when the strains carried R46, but the plasmid had no effect on the spontaneous reversion frequency of some other his missense mutations or of reversion rate of his frame-shift mutants (except for two uvrB derivatives of one single-base insertion mutant). The plasmid increased the ability of wild type, polA, and uvrB hosts to support plaque production by uv-irradiated phage, and made strain LT2 his G46 less sensitive to methyl methane sulfonate and to x rays and more responsive to the mutagenic effect of visible-light irradiation. R46 increased spontaneous reversion frequency of a his(amber) rec + strain, but had no such effect in its recA sublines. Since the plasmid in the absence of host recA function fails to produce its mutator effect, or to confer uv protection or to enhance uv mutagenesis, these three effects may be produced via some mechanism involved in recA-dependent deoxyribonucleic acid repair, perhaps by an increase in activity of the ''error-prone'' component of the inducible repair pathway

  7. Semiautomated Segmentation and Measurement of Cytoplasmic Vacuoles in a Neutrophil With General-Purpose Image Analysis Software.

    Science.gov (United States)

    Mizukami, Maki; Yamada, Misaki; Fukui, Sayaka; Fujimoto, Nao; Yoshida, Shigeru; Kaga, Sanae; Obata, Keiko; Jin, Shigeki; Miwa, Keiko; Masauzi, Nobuo

    2016-11-01

    Morphological observation of blood or marrow film is still described nonquantitatively. We developed a semiautomatic method for segmenting vacuoles from the cytoplasm using Photoshop (PS) and Image-J (IJ), called PS-IJ, and measured the relative entire cell area (rECA) and relative areas of vacuoles (rAV) in the cytoplasm of neutrophil with PS-IJ. Whole-blood samples were stored at 4°C with ethylenediaminetetraacetate and in two different preserving manners (P1 and P2). Color-tone intensity levels of neutrophil images were semiautomatically compensated using PS, and then vacuole portions were automatically segmented by IJ. The rAV and rECA were measured by counting pixels by IJ. For evaluating the accuracy in segmentations of vacuoles with PS-IJ, the rAV/rECA ratios calculated with results from PS-IJ were compared with those calculated with human eye and IJ (HE-IJ). The rECA and rAV/ in P1 significantly (P < 0.05, P < 0.05) were enlarged and increased, but did not significantly (P = 0.46, P = 0.21) change in P2. The rAV/rECA ratios by PS-IJ were significantly correlated (r = 0.90, P < 0.01) with those by HE-IJ. PS-IJ method can successfully segment vacuoles and measure the rAV and rECA, becoming a useful tool for quantitative description of morphological observation of blood and marrow film. © 2016 Wiley Periodicals, Inc.

  8. Effect of the mutations recB21, recD1013 and recJ284 of Escherichia Coli on the indirect recombinogenesis of the lambda bacteriophage; Efecto de las mutaciones recB21, recD1013 y recJ284 de Escherichia Coli sobre la recombinogenesis indirecta del bacteriofago lambda

    Energy Technology Data Exchange (ETDEWEB)

    Alcantara D, D. [ININ, 52045 Ocoyoacac, Estado de Mexico (Mexico)

    1994-01-15

    In this report its are related the indirect recombinogenesis of the lambda bacteriophage which depends on it happens in the guest cell after the UV irradiation with those cellular responses to the DNA damages and with the bacterial genes that intervene in them (one of those is the SOS response, controlled by the genes lexA and recA). However it has not been possible to establish a precise relationship among those two phenomena because contradictory results exist. (Author)

  9. Expression of the bacteriophage T4 denV structural gene in Escherichia coli

    International Nuclear Information System (INIS)

    Recinos, A. III; Augustine, M.L.; Higgins, K.M.; Lloyd, R.S.

    1986-01-01

    The expression of the T4 denV gene, which previously had been cloned in plasmid constructs downstream of the bacteriophage lambda hybrid promoter-operator oLpR, was analyzed under a variety of growth parameters. Expression of the denV gene product, endonuclease V, was confirmed in DNA repair-deficient Escherichia coli (uvrA recA) by Western blot analyses and by enhancements of resistance to UV irradiation

  10. Chromogenic method for rapid isolation of recA-like mutants of gram-negative bacteria.

    OpenAIRE

    Barbe, J; Fernandez de Henestrosa, A R; Calero, S; Gibert, I

    1991-01-01

    We have devised a rapid and widely applicable color test for detecting recA-like mutants of gram-negative bacteria. The technique depends on decreased expression of an Escherichia coli recA-lacZ fusion in recA mutants and uses a broad-host-range plasmid to transfer the fusion gene into new species. We describe the isolation of a recA-like mutant of Pseudomonas syringae by this technique.

  11. Rapid and apparently error-prone excision repair of nonreplicating UV-irradiated plasmids in Xenopus laevis oocytes.

    OpenAIRE

    Hays, J B; Ackerman, E J; Pang, Q S

    1990-01-01

    Repair of UV-irradiated plasmid DNA microinjected into frog oocytes was measured by two techniques: transformation of repair-deficient (delta uvrB delta recA delta phr) bacteria, and removal of UV endonuclease-sensitive sites (ESS). Transformation efficiencies relative to unirradiated plasmids were used to estimate the number of lethal lesions; the latter were assumed to be Poisson distributed. These estimates were in good agreement with measurements of ESS. By both criteria, plasmid DNA was ...

  12. Differential expression of SOS genes in an E. coli mutant producing unstable lexA protein enhances excision repair but inhibits mutagenesis

    International Nuclear Information System (INIS)

    Peterson, K.R.; Ganesan, A.K.; Mount, D.W.; Stanford Univ., CA)

    1986-01-01

    The SOS response is displayed following treatments which damage DNA or inhibit DNA replication. Two associated activities include enhanced capacity for DNA repair resulting from derepression of the recA, uvrA, uvrB and uvrD genes and increased mutagenesis due to derepression of recA, umuC and umuD. These changes are the consequence of the derepression of at least seventeen unlinked operons negatively regulated by LexA repressor. Following treatments that induce the SOS response, a signal molecule interacts with RecA protein, converting it to an activated form. Activated RecA protein facilitates the proteolytic cleavage of LexA repressor, which results in derepression of the regulon. The cell then enters a new physiological state during which time DNA repair processes are augmented. The lexA41 mutant of E. coli is a uv-resistant derivative of another mutant, lexA3, which produces a repressor that is not cleaved following inducing treatments. The resultant protein is unstable. Lac operon fusions to most of the genes in the SOS regulon were used to show that the various damage-inducible genes were derepressed to different extents. uvrA, B, and D were almost fully derepressed. Consistent with this finding, the rate of removal of T4 endonuclease V-sensitive sites was more rapid in the uv-irradiated lexA41 mutant than in normal cells, suggesting a more active excision repair system. We propose that the instability of the LexA41 protein reduces the intracellular concentration of repressor to a level that allows a high level of excision repair. The additional observation that SOS mutagenesis was only weakly induced in a lexA41 uvrA - mutant implies that the mutant protein partially represses one or more genes whose products promote SOS mutagenesis. 17 refs., 4 figs., 1 tab

  13. Effect of the mutations recB21, recD1013 and recJ284 of Escherichia Coli on the indirect recombinogenesis of the lambda bacteriophage

    International Nuclear Information System (INIS)

    Alcantara D, D.

    1994-01-01

    In this report its are related the indirect recombinogenesis of the lambda bacteriophage which depends on it happens in the guest cell after the UV irradiation with those cellular responses to the DNA damages and with the bacterial genes that intervene in them (one of those is the SOS response, controlled by the genes lexA and recA). However it has not been possible to establish a precise relationship among those two phenomena because contradictory results exist. (Author)

  14. The Effects of Aging and IQ on Item and Associative Memory

    OpenAIRE

    Ratcliff, Roger; Thapar, Anjali; McKoon, Gail

    2011-01-01

    The effects of aging and IQ on performance were examined in four memory tasks: item recognition, associative recognition, cued recall, and free recall. For item and associative recognition, accuracy and the response time distributions for correct and error responses were explained by Ratcliff’s (1978) diffusion model, at the level of individual participants. The values of the components of processing identified by the model for the recognition tasks, as well as accuracy for cued and free reca...

  15. Induction of genetic recombination in the lambda bacteriophage by ultraviolet radiation of the Escherichia Coli cells; Induccion de recombinacion genetica en el bacteriofago lambda por irradiacion ultravioleta de las cellulas de Escherichia Coli

    Energy Technology Data Exchange (ETDEWEB)

    Alcantara D, D. [ININ, 52045 Ocoyoacac, Estado de Mexico (Mexico)

    1986-12-15

    In this work there are reported the results that show that although the stimulation of the recombination of the Lambda bacteriophage, by UV irradiation of the cells of Escherichia Coli, it looks to be the result of the high expression of the functions of the SOS system, doesn't keep some relationship with the high concentration of protein reached RecA. (Author)

  16. Recombination-deficient mutant of Streptococcus faecalis

    International Nuclear Information System (INIS)

    Yagi, Y.; Clewell, D.B.

    1980-01-01

    An ultraviolet radiation-sensitive derivative of Streptococcus faecalis strain JH2-2 was isolated and found to be deficient in recombination, using a plasmid-plasmid recombination system. The strain was sensitive to chemical agents which interact with deoxyribonucleic acid and also underwent deoxyribonucleic acid degradation after ultraviolet irradiation. Thus, the mutant has properties similar to those of recA strains of Escherichia coli

  17. Environmental and molecular characterization of systems which affect genome alteration in pseudomonas aeruginosa

    International Nuclear Information System (INIS)

    Miller, R.V.; Kokjohn, T.A.; Sayler, G.S.

    1990-01-01

    Pseudomonas aeruginosa is used as a model organism to study genome alteration in freshwater microbial populations and horizontal gene transmission by both transduction and conjugation has been demonstrated. The studies have also provided data which suggest that intracellular genome instability may be increased in the aquatic environment as a result of stresses encountered by the cell in this habitat. The role of the P. aeruginosa recA analog in regulating genome instability is also addressed

  18. Model-based automatic tuning of a filtration control system for submerged anaerobic membrane bioreactors (AnMBR)

    OpenAIRE

    Robles Martínez, Ángel; Ruano García, María Victoria; Ribes Bertomeu, José; SECO TORRECILLAS, AURORA; FERRER, J.

    2014-01-01

    This paper describes a model-based method to optimise filtration in submerged AnMBRs. The method is applied to an advanced knowledge-based control system and considers three statistical methods: (1) sensitivity analysis (Morris screening method) to identify an input subset for the advanced controller; (2) Monte Carlo method (trajectory-based random sampling) to find suitable initial values for the control inputs; and (3) optimisation algorithm (performing as a supervisory controller) to re-ca...

  19. Antibiotics-free stable polyhydroxyalkanoate (PHA) production from carbon dioxide by recombinant cyanobacteria.

    Science.gov (United States)

    Akiyama, Hideo; Okuhata, Hiroshi; Onizuka, Takuo; Kanai, Shozo; Hirano, Masahiko; Tanaka, Satoshi; Sasaki, Ken; Miyasaka, Hitoshi

    2011-12-01

    A practical antibiotics-free plasmid expression system in cyanobacteria was developed by using the complementation of cyanobacterial recA null mutation with the EscherichiacolirecA gene on the plasmid. This system was applied to the production of polyhydroxyalkanoate (PHA), a biodegradable plastic, and the transgenic cyanobacteria stably maintained the pha genes for PHA production in the antibiotics-free medium, and accumulated up to 52% cell dry weight of PHA. Copyright © 2011 Elsevier Ltd. All rights reserved.

  20. 2-amino-purine, base analogue of adenine, induces in E. coli an error free repair acting upon UV lesions in doubles-stranded DNA

    International Nuclear Information System (INIS)

    Maenhaut-Michel, G.; Caillet-Fauquet, P.

    1982-01-01

    The survival of ultraviolet light (UV) -irradiated lambda phage is increased when host bacteria are grown in the presence of the base analogue 2-amino-purine (2 AP) prior infection. This increase in survival, which we call ''2 AP-reactivation'', has the following characteristics: it is not accompanied by mutagenesis; it occurs in recA - and lexA - bacterial mutants; is abolished in the polA - mutant (deficient in DNA polymerase I) [fr

  1. Identification of Burkholderia pseudomallei Near-Neighbor Species in the Northern Territory of Australia.

    Directory of Open Access Journals (Sweden)

    Jennifer L Ginther

    Full Text Available Identification and characterization of near-neighbor species are critical to the development of robust molecular diagnostic tools for biothreat agents. One such agent, Burkholderia pseudomallei, a soil bacterium and the causative agent of melioidosis, is lacking in this area because of its genomic diversity and widespread geographic distribution. The Burkholderia genus contains over 60 species and occupies a large range of environments including soil, plants, rhizospheres, water, animals and humans. The identification of novel species in new locations necessitates the need to identify the true global distribution of Burkholderia species, especially the members that are closely related to B. pseudomallei. In our current study, we used the Burkholderia-specific recA sequencing assay to analyze environmental samples from the Darwin region in the Northern Territory of Australia where melioidosis is endemic. Burkholderia recA PCR negative samples were further characterized using 16s rRNA sequencing for species identification. Phylogenetic analysis demonstrated that over 70% of the bacterial isolates were identified as B. ubonensis indicating that this species is common in the soil where B. pseudomallei is endemic. Bayesian phylogenetic analysis reveals many novel branches within the B. cepacia complex, one novel B. oklahomensis-like species, and one novel branch containing one isolate that is distinct from all other samples on the phylogenetic tree. During the analysis with recA sequencing, we discovered 2 single nucleotide polymorphisms in the reverse priming region of B. oklahomensis. A degenerate primer was developed and is proposed for future use. We conclude that the recA sequencing technique is an effective tool to classify Burkholderia and identify soil organisms in a melioidosis endemic area.

  2. Mandibular Symphyseal/Parasymphyseal Fracture with Incisor Tooth Loss: Preventing Lower Arch Constriction

    OpenAIRE

    Rahpeyma, Amin; Khajehahmadi, Saeedeh; Abdollahpour, Somayeh

    2015-01-01

    Mandibular fractures are the second most common fractures of the face after the nasal bone. Mandibular symphyseal/parasymphyseal fracture comprises 15.6 to 29.3% of mandibular fractures. Tooth loss in the fracture line is a known phenomenon, but space loss has not been evaluated comprehensively in the literature. In a retrospective study, patients with mandibular symphyseal/parasymphyseal fractures, who had been treated from 2012 to 2013 in Mashhad University, Iran, Emdadi Hospital, were reca...

  3. Cold Spring Harbor symposia on quantitative biology: Volume 49, Recombination at the DNA level

    International Nuclear Information System (INIS)

    1984-01-01

    This volume contains full papers prepared by the participants to the 1984 Cold Springs Harbor Symposia on Quantitative Biology. This year's theme is entitled Recombination at the DNA level. The volume consists of 93 articles grouped into subject areas entitled chromosome mechanics, yeast systems, mammalian homologous recombination, transposons, mu, plant transposons/T4 recombination, topoisomerase, resolvase and gyrase, Escherichia coli general recombination, RecA, repair, leukaryotic enzymes, integration and excision of bacteriophage, site-specific recombination, and recombination in vitro

  4. Differences in mutagenic and recombinational DNA repair in enterobacteria

    International Nuclear Information System (INIS)

    Sedgwick, S.G.; Goodwin, P.A.

    1985-01-01

    The incidence of recombinational DNA repair and inducible mutagenic DNA repair has been examined in Escherichia coli and 11 related species of enterobacteria. Recombinational repair was found to be a common feature of the DNA repair repertoire of at least 6 genera of enterobacteria. This conclusion is based on observations of (i) damage-induced synthesis of RecA-like proteins, (ii) nucleotide hybridization between E. coli recA sequences and some chromosomal DNAs, and (iii) recA-negative complementation by plasmids showing SOS-inducible expression of truncated E. coli recA genes. The mechanism of DNA damage-induced gene expression is therefore sufficiently conserved to allow non-E. coli regulatory elements to govern expression of these cloned truncated E. coli recA genes. In contrast, the process of mutagenic repair, which uses umuC+ umuD+ gene products in E. coli, appeared less widespread. Little ultraviolet light-induced mutagenesis to rifampicin resistance was detected outside the genus Escherichia, and even within the genus induced mutagenesis was detected in only 3 out of 6 species. Nucleotide hybridization showed that sequences like the E. coli umuCD+ gene are not found in these poorly mutable organisms. Evolutionary questions raised by the sporadic incidence of inducible mutagenic repair are discussed

  5. SOS regulatory elements are essential for UPEC pathogenesis.

    Science.gov (United States)

    Li, Birong; Smith, Peter; Horvath, Dennis J; Romesberg, Floyd E; Justice, Sheryl S

    2010-08-01

    Epithelial cells are highly regarded as the first line of defense against microorganisms, but the mechanisms used to control bacterial diseases are poorly understood. A component of the DNA damage repair regulon, SulA, is essential for UPEC virulence in a mouse model for human urinary tract infection, suggesting that DNA damage is a key mediator in the primary control of pathogens within the epithelium. In this study, we examine the role of DNA damage repair regulators in the intracellular lifestyle of UPEC within superficial bladder epithelial cells. LexA and RecA coordinate various operons for repair of DNA damage due to exogenous and endogenous agents and are known regulators of sulA. UPEC strains defective in regulation of the SOS response mediated by RecA and LexA display attenuated virulence in immunocompetent mice within the first 6 h post infection. RecA and LexA regulation of the SOS regulon is dispensable in immunocompromised mice. These data suggest that epithelial cells produce sufficient levels of DNA damaging agents, such that the bacterial DNA damage repair response is essential, as a means to control invading bacteria. Since many pathogens interact with the epithelium before exposure to professional phagocytes, it is likely that adaptation to oxidative radicals during intracellular growth provides additional protection from killing by innate immune phagocytes. Copyright 2010 Elsevier Masson SAS. All rights reserved.

  6. Stabilized gene duplication enables long-term selection-free heterologous pathway expression.

    Science.gov (United States)

    Tyo, Keith E J; Ajikumar, Parayil Kumaran; Stephanopoulos, Gregory

    2009-08-01

    Engineering robust microbes for the biotech industry typically requires high-level, genetically stable expression of heterologous genes and pathways. Although plasmids have been used for this task, fundamental issues concerning their genetic stability have not been adequately addressed. Here we describe chemically inducible chromosomal evolution (CIChE), a plasmid-free, high gene copy expression system for engineering Escherichia coli. CIChE uses E. coli recA homologous recombination to evolve a chromosome with approximately 40 consecutive copies of a recombinant pathway. Pathway copy number is stabilized by recA knockout, and the resulting engineered strain requires no selection markers and is unaffected by plasmid instabilities. Comparison of CIChE-engineered strains with equivalent plasmids revealed that CIChE improved genetic stability approximately tenfold and growth phase-specific productivity approximately fourfold for a strain producing the high metabolic burden-biopolymer poly-3-hydroxybutyrate. We also increased the yield of the nutraceutical lycopene by 60%. CIChE should be applicable in many organisms, as it only requires having targeted genomic integration methods and a recA homolog.

  7. Mycobacterium smegmatis PafBC is involved in regulation of DNA damage response.

    Science.gov (United States)

    Fudrini Olivencia, Begonia; Müller, Andreas U; Roschitzki, Bernd; Burger, Sibylle; Weber-Ban, Eilika; Imkamp, Frank

    2017-10-25

    Two genes, pafB and pafC, are organized in an operon with the Pup-ligase gene pafA, which is part of the Pup-proteasome system (PPS) present in mycobacteria and other actinobacteria. The PPS is crucial for Mycobacterium tuberculosis resistance towards reactive nitrogen intermediates (RNI). However, pafB and pafC apparently play only a minor role in RNI resistance. To characterize their function, we generated a pafBC deletion in Mycobacterium smegmatis (Msm). Proteome analysis of the mutant strain revealed decreased cellular levels of various proteins involved in DNA damage repair, including recombinase A (RecA). In agreement with this finding, Msm ΔpafBC displayed increased sensitivity to DNA damaging agents. In mycobacteria two pathways regulate DNA repair genes: the LexA/RecA-dependent SOS response and a predominant pathway that controls gene expression via a LexA/RecA-independent promoter, termed P1. PafB and PafC feature winged helix-turn-helix DNA binding motifs and we demonstrate that together they form a stable heterodimer in vitro, implying a function as a heterodimeric transcriptional regulator. Indeed, P1-driven transcription of recA was decreased in Msm ΔpafBC under standard conditions and induction of recA expression upon DNA damage was strongly impaired. Taken together, our data indicate an important regulatory function of PafBC in the mycobacterial DNA damage response.

  8. Aspectos psicológicos da recidiva em Oncologia Pediátrica: uma revisão integrativa

    Directory of Open Access Journals (Sweden)

    Marina Noronha Ferraz de Arruda-Colli

    2015-01-01

    Full Text Available Este estudio tuvo como objetivo comprender cómo los aspectos psicológicos de la recaída del cáncer infantil han sido investigados en la literatura científica. Se realizó una revisión integradora, con la búsqueda electrónica a partir de las bases de indexa - ción PubMed, LILACS, SciELO, PsycINFO y Cochrane Library, a partir de los siguientes descriptores: neoplasias, recaída, recaída del cáncer, niño, familia, padres, psicolo - gía infantil. Se plantearon 473 artículos. Luego de la aplicación de los criterios de inclusión/exclusión, 14 publicaciones permanecieron en la selección final. Se eviden - cia la predominancia de los delineamientos empíricos con enfoque cuantitativo. Las principales estrategias de afrontamiento utilizadas por los pacientes y sus cuidado - res, y las intervenciones que promuevan la calidad de vida son descritos. Una visión panorámica del campo de la investigación evidencia la necesidad de hacer frente a la incertidumbre y organizarse para afrontar el reinicio del tratamiento. Comprender estas variables puede auxiliar a la organización de la atención integral a la familia.

  9. Molecular genetic and biochemical analyses of a DNA repair gene from Serratia marcescens

    International Nuclear Information System (INIS)

    Murphy, K.E.

    1989-01-01

    In Escherichia coli, the SOS response and two 3-methyladenine DNA glycosylases (TagI and TagII) are required for repair of DNA damaged by alkylating agents such as methyl methanesulfonate (MMS). Mutations of the recA gene eliminate the SOS response. TagI and TagII are encoded by the tag and alkA genes, respectively. A gene (rpr) encoding 3-methyladenine DNA glycosylase activity was isolated from the Gram-negative bacterium Serratia marcescens. The gene, localized to a 1.5-kilobase pair SmaI-HindIII restriction fragment, was cloned into plasmid pUC18. The clone complemented E. coli tag alkA and recA mutations for MMS resistance. The rpr gene did not, however, complement recA mutations for resistance to ultraviolet light or the ability to perform homologous recombination reactions, nor did it complement E. coli ada or alkB mutations. Two proteins of molecular weights 42,000 and 16,000 were produced from the rpr locus. Analysis of deletion and insertion mutants of rpr suggested that the 42kD molecule is the active protein. The 16kD protein may either be a breakdown product of the 42kD species or may be encoded by another gene overlapping the reading frame of the rpr gene. Biochemical assays showed that the rpr gene product (Rpr) possesses 3-methyladenine DNA glycosylase activity

  10. The lon gene and photoprotection in Escherichia coli K-12

    International Nuclear Information System (INIS)

    Waksman, G.; Thomas, G.; Favre, A.

    1984-01-01

    Photoprotection, i.e. the increased resistance of the cells preilluminated with near ultraviolet light (300-380 nm) to the lethal action of 254nm radiations requires either an integrated prophage or a recA mutation in Escherichia coli K12 strains. Significant photoprotection occurs in an Escherichia coli K12 recA + cell containing the lon allele responsible for filamentous growth after 254nm irradiation. The Fil phenotype can be suppressed by the sfiA or sfiB suppressor genes. Since the E. coli K12 recA + lon sfiB strain exhibits no more photoprotection, it is concluded that in lon strains photoprotection is due to the abolition of the 254nm induced filamentation by the near ultraviolet treatment. In addition, near ultraviolet illumination of the cells leads to a severe restriction of the bulk protein synthesis. This effect is observed only in nuv + cells that contain 4-thiouridine the chromophore responsible for photoprotection. It is proposed that in lon (lysogenic strains) photoprotection is due to prevention of the SOS response. During the growth lag, the low residual level of protein synthesis does not allow the induction of the SOS response and accordingly prevents filamentation (the lytic cycle). (author)

  11. Developing single-molecule TPM experiments for direct observation of successful RecA-mediated strand exchange reaction.

    Science.gov (United States)

    Fan, Hsiu-Fang; Cox, Michael M; Li, Hung-Wen

    2011-01-01

    RecA recombinases play a central role in homologous recombination. Once assembled on single-stranded (ss) DNA, RecA nucleoprotein filaments mediate the pairing of homologous DNA sequences and strand exchange processes. We have designed two experiments based on tethered particle motion (TPM) to investigate the fates of the invading and the outgoing strands during E. coli RecA-mediated pairing and strand exchange at the single-molecule level in the absence of force. TPM experiments measure the tethered bead Brownian motion indicative of the DNA tether length change resulting from RecA binding and dissociation. Experiments with beads labeled on either the invading strand or the outgoing strand showed that DNA pairing and strand exchange occurs successfully in the presence of either ATP or its non-hydrolyzable analog, ATPγS. The strand exchange rates and efficiencies are similar under both ATP and ATPγS conditions. In addition, the Brownian motion time-courses suggest that the strand exchange process progresses uni-directionally in the 5'-to-3' fashion, using a synapse segment with a wide and continuous size distribution.

  12. UV-lysogenic induction of lambda phage in lexAl mutants of Escherichia coli: kinetics of the process

    Energy Technology Data Exchange (ETDEWEB)

    Carvalho, R.E.S.; Leitao, A.C. (Rio de Janeiro Univ. (Brazil). Inst. de Biofisica)

    1984-05-01

    Although the lex gene has been described as being required for lysogenic induction, both this work and the work of others have reported lambda prophage induction in some lexA mutants. However, the characteristics of the process were not defined. UV induction of prophage in a lexAl mutant is described at a slightly lower level and requiring 2 times longer than the wild type. As demonstrated in some work, in cells treated with low levels of rifampicin (RIF) no new synthesis of RecA protein is needed for the prophage induction although the onset of lysis is delayed. It is suggested that the lysogenic induction in lexA cells is due to the same mechanism that induces prophage in the wild type cells treated with RIF. That is, the induction is due to the cleavage of lambda repressor by the basal RecA protease in the DNA-single-strand gap, since RecA protease and monomer repressor both have high affinity for this type of DNA. So, LexA protein need not be cleaved for the prophage induction. No Weigle-reactivation (WR) was detected in the lex mutant even after a long post-irradiation incubation, suggesting that unlike prophage induction, WR requires LexA protein cleavage.

  13. Near-ultraviolet radiation blocks SOS responses to DNA damage in Escherichia coli

    Energy Technology Data Exchange (ETDEWEB)

    Turner, M.A.; Eisenstark, A.

    1984-01-01

    Escherichia coli cells in which the recA promoter is fused to a lac structural gene, (Mu) Mud(Ap,lac)::rec, were irradiated with two far-ultraviolet light wavelengths (254 and 290 nm), selected monochromatic near-ultraviolet (NUV) wavelengths 313 nm, 334 nm, 365 nm, or broad band solar-UV (290-420 nm) from a solar simulator. Irradiation with the two far-ultraviolet wavelengths was followed by high yields of ..beta..-galactosidase, lambda prophage induction, and Weigle reactivation. These end points were not observed after irradiation with the selected NUV wavelengths or the broad spectrum solar-UV. Thus, neither broad spectrum solar-UV nor monochromatic NUV wavelengths resulted in the derepression of the recA promoter. Further, prior exposure of the cells either to the selected monochromatic NUV wavelengths or to solar-UV inhibited a) the induction of ..beta..-galactosidase by subsequent 254-nm radiation, b) subsequent 254-nm induction of lambda prophage, c) Weigle reactivation, and d) mutation frequency. These observations are consistent with the hypothesis that NUV blocks subsequent recA protease action.

  14. Nucleotide cofactor-dependent structural change of Xenopus laevis Rad51 protein filament detected by small-angle neutron scattering measurements in solution

    DEFF Research Database (Denmark)

    Ellouze, C.; Kim, H.-K.; Maeshima, K.

    1997-01-01

    of RecA. Upon ATP binding under high-salt conditions (600 mM NaCl), the helical pitch of XRad51.1 filament was increased from 8 to 10 nm and the cross-sectional diameter decreased from 7 to 6 nm. The pitch sizes of XRad51.1 are similar to, though slightly larger than, those of RecA filament under......Rad51 protein, a eukaryotic homologue of RecA protein, forms a filamentous complex with DNA and catalyzes homologous recombination. We have analyzed the structure of Xenopus Rad51 protein (XRad51.1) in solution by small-angle neutron scattering (SANS). The measurements showed that XRad51.1 forms...... a helical filament independently of DNA. The sizes of the cross-sectional and helical pitch of the filament could be determined, respectively, from a Guinier plot and the position of the subsidiary maximum of SANS data. We observed that the helical structure is modified by nucleotide binding as in the case...

  15. Biodetection of potential genotoxic pollutants entering the human food chain through ashes used in livestock diets.

    Science.gov (United States)

    Sanchez-Vicente, Laura; Herraez, Elisa; Briz, Oscar; Nogales, Rogelio; Molina-Alcaide, Eduarda; Marin, Jose J G

    2016-08-15

    Ash derived from energy generation is used as a source of minerals in livestock feeds. The microbial biosensor recApr-Luc2 was built to detect genotoxic hazard in recycled ash. Escherichia coli SOS gene (recA, lexA, dinI and umuC) expression in response to cisplatin-induced DNA damage led to the selection of the recA promoter. The biosensor required functional RecA expression to respond to genotoxic heavy metals (Cr>Cd≈Pb), and polluted ash induced a strong recApr-Luc2 response. In human liver and intestinal cells, heavy metals induced acute toxicity (Cr>Cd>Pb) at concentrations sufficient to activate recApr-Luc2. Cytostatic effects, including genotoxicity, were cell- and metal-dependent, apart from Cr. In agreement with the recApr-Luc2 bioassay, Cr had the strongest effect in all cells. In conclusion, recApr-Luc2 could be useful for evaluating the genotoxic risk of pollutants present in ash that might be concentrated in animal products and, thus, entering the human food chain. Copyright © 2016 Elsevier Ltd. All rights reserved.

  16. Verificação, em condiçoes naturais, da infecção de mais três hematófagos (Anophelineos, Flebótomos e Simulídeos em leprosos Verification, in natural conditions, of infection of more three hematophagi (Anopheline, Flebotomus and Simulidae in lepers

    Directory of Open Access Journals (Sweden)

    H. C. de Souza-Araujo

    1943-10-01

    Full Text Available The A. went last August to the State of Minas Gerais to continue his studies on transmission of leprosy by insects. He selected ten lepers (all L3 cases for his experiments. It happened in the middle of August, a few day after freezing temperature. Practically there were no day mosquitoes, even near a river where in March there were very many. Bringing the patients to a wood, near the Peixe River (Fish River, at the dawn they were attacked by very many Anopheles, some flebotomus, a few Simulium and very rare Culex scapularis. All these insects became infected, in different degrees, by biting such patients. CONCLUSIONS. 1. Two species of Anopheles (A. albitarsis and A. tarsimaculatus became strongly infected by Hansen bacilli. 2. By dissection done by Dr. Oliveira Castro were found lepra bacilli in various points of the proboscis of two Flebotomus (F. intermedius and in their stomachs. 3. By smearing were found lepra bacilli in two specimens of Simulium sp. (probably pertinax. 4. It was confirmed also the verification done last March, at the same Leper Colony, that Phthirius pubis can be also a carrier of lepra bacillus. 5. There was confirmed also natural infection of nymphs of Amblyomma cajennense in lepers. Dr. Oliveira Castro is dissecting the Anopheles to locate the bacilli in their organisms and he started, with the cooperation of the Director of Colonia Santa Fé, Dr. José Mariano, attenpts to re-infect a group of negative-nerve cases of leprosy with infected mosquitoes.

  17. PCR-based techniques for leprosy diagnosis: from the laboratory to the clinic.

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    Alejandra Nóbrega Martinez

    2014-04-01

    Full Text Available In leprosy, classic diagnostic tools based on bacillary counts and histopathology have been facing hurdles, especially in distinguishing latent infection from active disease and diagnosing paucibacillary clinical forms. Serological tests and IFN-gamma releasing assays (IGRA that employ humoral and cellular immune parameters, respectively, are also being used, but recent results indicate that quantitative PCR (qPCR is a key technique due to its higher sensitivity and specificity. In fact, advances concerning the structure and function of the Mycobacterium leprae genome led to the development of specific PCR-based gene amplification assays for leprosy diagnosis and monitoring of household contacts. Also, based on the validation of point-of-care technologies for M. tuberculosis DNA detection, it is clear that the same advantages of rapid DNA detection could be observed in respect to leprosy. So far, PCR has proven useful in the determination of transmission routes, M. leprae viability, and drug resistance in leprosy. However, PCR has been ascertained to be especially valuable in diagnosing difficult cases like pure neural leprosy (PNL, paucibacillary (PB, and patients with atypical clinical presentation and histopathological features compatible with leprosy. Also, the detection of M. leprae DNA in different samples of the household contacts of leprosy patients is very promising. Although a positive PCR result is not sufficient to establish a causal relationship with disease outcome, quantitation provided by qPCR is clearly capable of indicating increased risk of developing the disease and could alert clinicians to follow these contacts more closely or even define rules for chemoprophylaxis.

  18. The challenge of producing skin test antigens with minimal resources suitable for human application against a neglected tropical disease; leprosy.

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    Becky L Rivoire

    Full Text Available True incidence of leprosy and its impact on transmission will not be understood until a tool is available to measure pre-symptomatic infection. Diagnosis of leprosy disease is currently based on clinical symptoms, which on average take 3-10 years to manifest. The fact that incidence, as defined by new case detection, equates with prevalence, i.e., registered cases, suggests that the cycle of transmission has not been fully intercepted by implementation of multiple drug therapy. This is supported by a high incidence of childhood leprosy. Epidemiological screening for pre-symptomatic leprosy in large endemic populations is required to facilitate targeted chemoprophylactic interventions. Such a test must be sensitive, specific, simple to administer, cost-effective, and easy to interpret. The intradermal skin test method that measures cell-mediated immunity was explored as the best option. Prior knowledge on skin testing of healthy subjects and leprosy patients with whole or partially fractionated Mycobacterium leprae bacilli, such as Lepromin or the Rees' or Convit' antigens, has established an acceptable safety and potency profile of these antigens. These data, along with immunoreactivity data, laid the foundation for two new leprosy skin test antigens, MLSA-LAM (M. leprae soluble antigen devoid of mycobacterial lipoglycans, primarily lipoarabinomannan and MLCwA (M. leprae cell wall antigens. In the absence of commercial interest, the challenge was to develop these antigens under current good manufacturing practices in an acceptable local pilot facility and submit an Investigational New Drug to the Food and Drug Administration to allow a first-in-human phase I clinical trial.

  19. Relative entropy differences in bacterial chromosomes, plasmids, phages and genomic islands

    DEFF Research Database (Denmark)

    Bohlin, Jon; van Passel, Mark W. J.; Snipen, Lars

    2012-01-01

    , and plasmids. Relative entropy was estimated using the Kullback-Leibler measure. Results: Relative entropy was highest in bacterial chromosomes and had the sequence chromosomes > GI > phage > plasmid. There was an association between relative entropy and AT content in chromosomes, phages, plasmids and GIs...... with the strongest association being in phages. Relative entropy was also found to be lower in the obligate intracellular Mycobacterium leprae than in the related M. tuberculosis when measured on a shared set of highly conserved genes. Conclusions: We argue that relative entropy differences reflect how plasmids...... chromosomes and stably incorporated GIs compared to the transient or independent replicons such as phages and plasmids....

  20. DNAhsp65 Vaccine as Therapy against Paracoccidioidomycosis.

    Science.gov (United States)

    Ribeiro, Alice M; Amaral, André C; Felipe, Maria Sueli S; Bocca, Anamelia L

    2017-01-01

    The conventional treatment for fungal diseases usually shows long periods of therapy and the high frequency of relapses and sequels. New strategies of the treatment are necessary. We have shown that the Mycobacterium leprae HSP65 gene can be successfully used as therapy against murine Paracoccidioidomycosis (PCM). Here, we described the methodology of DNAhsp65 immunotherapy in mice infected with the dimorphic fungus Paracoccidioides brasiliensis, one of PCM agent, evaluating cytokines levels, fungal burden, and lung injury. Our results provide a new prospective on the immunotherapy of mycosis.

  1. Pesquisa documental sobre a história da hanseníase no Brasil

    OpenAIRE

    Santos,Vicente Saul Moreira dos

    2003-01-01

    Este trabalho apresenta resultados parciais de uma pesquisa sobre fontes relativas à lepra, realizada em diversas instituições da cidade do Rio de Janeiro. No Real Gabinete Português de Leitura, no Arquivo Nacional e na Biblioteca Nacional foram pesquisados livros, índices, ofícios e fotografias sobre administração de leprosários e artigos sobre o tratamento da doença. No Centro de Pesquisa e Documentação de História Contemporânea do Brasil (CPDOC-FGV) foram consultados diversos arquivos, des...

  2. Sorologia da hanseníase utilizando PGL-I: revisão sistemática Leprosy serology using PGL-I: a systematic review

    OpenAIRE

    Rodrigo Scaliante de Moura; Karla Lucena Calado; Maria Leide W. Oliveira; Samira Bührer-Sékula

    2008-01-01

    A sorologia utilizando o antígeno espécie-específico do Mycobacterium leprae, PGL-I, pode ser um marcador de carga bacteriana em pacientes com hanseníase. Estudos identificaram potencial de uso da sorologia na classificação de pacientes para fins de tratamento, monitoramento de terapia, risco de recidiva e na seleção dos contatos com maior risco de adoecer. Foi realizada uma revisão sistemática e 26 artigos foram incluídos na análise comparativa. Avaliamos os resultados do uso da sorologia PG...

  3. Sorologia da hanseníase utilizando PGL-I: revisão sistemática

    OpenAIRE

    Moura,Rodrigo Scaliante de; Calado,Karla Lucena; Oliveira,Maria Leide W.; Bührer-Sékula,Samira

    2008-01-01

    A sorologia utilizando o antígeno espécie-específico do Mycobacterium leprae, PGL-I, pode ser um marcador de carga bacteriana em pacientes com hanseníase. Estudos identificaram potencial de uso da sorologia na classificação de pacientes para fins de tratamento, monitoramento de terapia, risco de recidiva e na seleção dos contatos com maior risco de adoecer. Foi realizada uma revisão sistemática e 26 artigos foram incluídos na análise comparativa. Avaliamos os resultados do uso da sorologia PG...

  4. Association analysis of the genetic polymorphisms with leprosy subtypes in Chinese Han population from Northern China.

    Science.gov (United States)

    Wang, Chuan; Wang, Zhenzhen; Wang, Honglei; Pan, Qing; Fu, Xi'an; Liu, Tingting; Yu, Gongqi; Liu, Hong; Zhang, Furen

    2018-03-24

    Leprosy is characterized by a broad spectrum of clinical manifestations that extend from paucibacillary (PB) to multibacillary (MB) depending upon the different immunologic response to the invading of M. leprae 3,4 . It has been widely accepted that genetic predisposition plays the crucial role in the different clinical manifestations. Host susceptibility to leprosy is modified by number of genes via two stages. In the first stage, a group of genes confers susceptibility to leprosy per se; A second group of genes are associated to the type of host immune response and leprosy subtype 5 . This article is protected by copyright. All rights reserved. This article is protected by copyright. All rights reserved.

  5. IL-10 and NOS2 modulate antigen-specific reactivity and nerve infiltration by T cells in experimental leprosy.

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    Deanna A Hagge

    2014-09-01

    Full Text Available Although immunopathology dictates clinical outcome in leprosy, the dynamics of early and chronic infection are poorly defined. In the tuberculoid region of the spectrum, Mycobacterium leprae growth is restricted yet a severe granulomatous lesion can occur. The evolution and maintenance of chronic inflammatory processes like those observed in the leprosy granuloma involve an ongoing network of communications via cytokines. IL-10 has immunosuppressive properties and IL-10 genetic variants have been associated with leprosy development and reactions.The role of IL-10 in resistance and inflammation in leprosy was investigated using Mycobacterium leprae infection of mice deficient in IL-10 (IL-10-/-, as well as mice deficient in both inducible nitric oxide synthase (NOS2-/- and IL-10 (10NOS2-/-. Although a lack of IL-10 did not affect M. leprae multiplication in the footpads (FP, inflammation increased from C57Bl/6 (B6leprae cell wall, membrane, and cytosol antigens and ML2028 (Ag85B were significantly increased in the evolved granuloma in NOS2-/- FP compared to B6 and IL-10-/- during early and peak phases. In 10NOS2-/- FP, CD4+CD44+ and especially CD8+CD44+ responses were augmented even further to these antigens as well as to ML0380 (GroES, ML2038 (bacterioferritin, and ML1877 (EF-Tu. Moreover, fragmented nerves containing CD4+ cells were present in 10NOS2-/- FP.The 10NOS2-/- strain offers insight on the regulation of granuloma formation and maintenance by immune modulators in the resistant forms of leprosy and presents a new model for investigating the pathogenesis of neurological involvement.

  6. Development of Hodgkin's disease in a patient with leprosy.

    Science.gov (United States)

    Weshler, Z; Leviatan, A; Gordon, R; Kopolovic, J

    1978-01-01

    We present a patient with leprosy who developed Hodgkin's disease of the nodular sclerosing type. There are two previous reports describing the combination of leprosy and Hodgkin's disease in a single patient [3, 9]. Hodgkin's disease was diagnosed 14 months after the complete disappearance of mycobacterium leprae from the skin lesions, under treatment with DDS (diamino-diphenyl-sulfone). Hodgkin's disease was treated by irradiation and chemotherapy. Obstructive jaundice developed which resolved under treatment by irradiation of the hilar area of the liver, chemotherapy and hormones. During two years of immuno-suppressive therapy, without DDS, no exacerbation of the leprosy occurred.

  7. Leprosy in Denmark 1980-2010

    DEFF Research Database (Denmark)

    Aftab, Huma; Nielsen, Susanne D.; Bygbjerg, Ib C.

    2016-01-01

    BACKGROUND: Leprosy, caused by Mycobacterium leprae, is a chronic and progressive granulomatous disease affecting mainly the skin and the peripheral nervous system. If left unrecognized, the infection can lead to permanent nerve damage and disability. The clinical presentation depends on the immune...... or with history of travel or work in the tropics. Due to the long incubation period with symptoms presenting long after immigration or return, clinicians often do not have the diagnosis in mind. The wide spectrum of symptoms and immunological reactions further complicates the diagnostic process. Treatment...

  8. Engineering protein scaffolds for protein separation, biocatalysis and nanotechnology applications

    Science.gov (United States)

    Liu, Fang

    Globally, there is growing appreciation for developing a sustainable economy that uses eco-efficient bio-processes. Biotechnology provides an increasing range of tools for industry to help reduce cost and improve environmental performance. Inspired by the naturally evolved machineries of protein scaffolds and their binding ligands, synthetic protein scaffolds were engineered based on cohesin-dockerin interactions and metal chelating peptides to tackle the challenges and make improvements in three specific areas: (1) protein purification, (2) biofuel cells, and (3) nanomaterial synthesis. The first objective was to develop efficient and cost-effective non-chromatographic purification processes to purify recombinant proteins in an effort to meet the dramatically growing market of protein drugs. In our design, the target protein was genetically fused with a dockerin domain from Clostridium thermocellum and direct purification and recovery was achieved using thermo-responsive elastin-like polypeptide (ELP) scaffold containing the cohesin domain from the same species. By exploiting the highly specific interaction between the dockerin and cohesin domain and the reversible aggregation property of ELP, highly purified and active dockerin-tagged proteins, such as endoglucanase CelA, chloramphenicol acetyl transferase (CAT) and enhanced green fluorescence protein (EGFP), were recovered directly from crude cell extracts in a single purification step with yields achieving over 90%. Incorporation of a self-cleaving intein domain enabled rapid removal of the affinity tag from the target proteins by another cycle of thermal precipitation. The purification cost can be further reduced by regenerating and recycling the ELP-cohesin capturing scaffolds. However, due to the high binding affinity between cohesin and dockerin domains, the bound dockerin-intein tag cannot be completely disassociated from ELP-cohesin scaffold after binding. Therefore, a truncated dockerin with the calcium

  9. Osteological, biomolecular and geochemical examination of an early anglo-saxon case of lepromatous leprosy.

    Science.gov (United States)

    Inskip, Sarah A; Taylor, G Michael; Zakrzewski, Sonia R; Mays, Simon A; Pike, Alistair W G; Llewellyn, Gareth; Williams, Christopher M; Lee, Oona Y-C; Wu, Houdini H T; Minnikin, David E; Besra, Gurdyal S; Stewart, Graham R

    2015-01-01

    We have examined a 5th to 6th century inhumation from Great Chesterford, Essex, UK. The incomplete remains are those of a young male, aged around 21-35 years at death. The remains show osteological evidence of lepromatous leprosy (LL) and this was confirmed by lipid biomarker analysis and ancient DNA (aDNA) analysis, which provided evidence for both multi-copy and single copy loci from the Mycobacterium leprae genome. Genotyping showed the strain belonged to the 3I lineage, but the Great Chesterford isolate appeared to be ancestral to 3I strains found in later medieval cases in southern Britain and also continental Europe. While a number of contemporaneous cases exist, at present, this case of leprosy is the earliest radiocarbon dated case in Britain confirmed by both aDNA and lipid biomarkers. Importantly, Strontium and Oxygen isotope analysis suggest that the individual is likely to have originated from outside Britain. This potentially sheds light on the origins of the strain in Britain and its subsequent spread to other parts of the world, including the Americas where the 3I lineage of M. leprae is still found in some southern states of America.

  10. Pure neuritic leprosy: Resolving diagnostic issues in acid fast bacilli (AFB)-negative nerve biopsies: A single centre experience from South India.

    Science.gov (United States)

    Hui, Monalisa; Uppin, Megha S; Challa, Sundaram; Meena, A K; Kaul, Subhash

    2015-01-01

    Demonstration of lepra bacilli is essential for definite or unequivocal diagnosis of pure neuritic leprosy (PNL) on nerve biopsy. However, nerves always do not show bacilli owing to the changes of previous therapy or due to low bacillary load in tuberculoid forms. In absence of granuloma or lepra bacilli, other morphologic changes in endoneurium and perineurium can be of help in making a probable diagnosis of PNL and treating the patient with multidrug therapy. Forty-six biopsies of PNL were retrospectively reviewed and histologic findings were compared with 25 biopsies of non leprosy neuropathies (NLN) including vasculitic neuropathy and chronic inflammatory demyelinating polyneuropathy (CIDP). The distribution of endoneurial infiltrate and fibrosis, perineurial thickening, and myelin abnormalities were compared between PNL and NLN biopsies and analyzed by Chi-square test. Out of 46 PNL casses, 24 (52.17 %) biopsies were negative for acid fast bacilli (AFB). In these cases, the features which favor a diagnosis of AFB-negative PNL were endoneurial infiltrate (51.1%), endoneurial fibrosis (54.2%), perineurial thickening (70.8%), and reduced number of myelinated nerve fibers (75%). Nerve biopsy is an efficient tool to diagnose PNL and differentiate it from other causes of NLN. In absence of AFB, the diagnosis of PNL is challenging. In this article, we have satisfactorily evaluated the various hisopthological features and found that endoneurial inflammation, dense fibrosis, and reduction in the number of myelinated nerve fibers are strong supportive indicators of PNL regardless of AFB positivity.

  11. Osteological, biomolecular and geochemical examination of an early anglo-saxon case of lepromatous leprosy.

    Directory of Open Access Journals (Sweden)

    Sarah A Inskip

    Full Text Available We have examined a 5th to 6th century inhumation from Great Chesterford, Essex, UK. The incomplete remains are those of a young male, aged around 21-35 years at death. The remains show osteological evidence of lepromatous leprosy (LL and this was confirmed by lipid biomarker analysis and ancient DNA (aDNA analysis, which provided evidence for both multi-copy and single copy loci from the Mycobacterium leprae genome. Genotyping showed the strain belonged to the 3I lineage, but the Great Chesterford isolate appeared to be ancestral to 3I strains found in later medieval cases in southern Britain and also continental Europe. While a number of contemporaneous cases exist, at present, this case of leprosy is the earliest radiocarbon dated case in Britain confirmed by both aDNA and lipid biomarkers. Importantly, Strontium and Oxygen isotope analysis suggest that the individual is likely to have originated from outside Britain. This potentially sheds light on the origins of the strain in Britain and its subsequent spread to other parts of the world, including the Americas where the 3I lineage of M. leprae is still found in some southern states of America.

  12. Marcas em si: vivenciando a dor do (auto preconceito

    Directory of Open Access Journals (Sweden)

    Iací Proença Palmeira

    2013-12-01

    Full Text Available Pesquisa qualitativa cujo objetivo foi compreender as representações sociais do corpo por mulheres com alterações provocadas pela hanseníase, para que os cuidados de enfermagem possam atender de forma mais abrangente às suas necessidades. O referencial foi o da teoria das representações sociais. Participaram quarenta e três mulheres de uma unidade de referência especializada em dermatologia sanitária. Realizaram-se entrevistas, cujos conteúdos foram processados pelo software ALCESTE, o qual gerou uma classe cujos vocábulos associaram-se ao preconceito ancorado na autopercepção negativa da hanseníase. O esquema explicativo das representações sociais mostrou uma forte marca da memória social da lepra, cuja construção simbólica ampara-se no medo da doença. Concluiu-se sobre a necessidade de se investir na desconstrução de crenças arcaicas sobre a lepra para que se tenham novos modos de agir frente à hanseníase.

  13. Decoding the similarities and differences among mycobacterial species.

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    Sony Malhotra

    2017-08-01

    Full Text Available Mycobacteriaceae comprises pathogenic species such as Mycobacterium tuberculosis, M. leprae and M. abscessus, as well as non-pathogenic species, for example, M. smegmatis and M. thermoresistibile. Genome comparison and annotation studies provide insights into genome evolutionary relatedness, identify unique and pathogenicity-related genes in each species, and explore new targets that could be used for developing new diagnostics and therapeutics. Here, we present a comparative analysis of ten-mycobacterial genomes with the objective of identifying similarities and differences between pathogenic and non-pathogenic species. We identified 1080 core orthologous clusters that were enriched in proteins involved in amino acid and purine/pyrimidine biosynthetic pathways, DNA-related processes (replication, transcription, recombination and repair, RNA-methylation and modification, and cell-wall polysaccharide biosynthetic pathways. For their pathogenicity and survival in the host cell, pathogenic species have gained specific sets of genes involved in repair and protection of their genomic DNA. M. leprae is of special interest owing to its smallest genome (1600 genes and ~1300 psuedogenes, yet poor genome annotation. More than 75% of the pseudogenes were found to have a functional ortholog in the other mycobacterial genomes and belong to protein families such as transferases, oxidoreductases and hydrolases.

  14. Reacción leprosa y sindrome eritema nudoso y E. polimorfo simultaneos con descamación eritrodermica generalizada

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    J. Ignacio Chala H.

    1939-11-01

    Full Text Available Antes de iniciar el estudio de las observaciones de R. L. de tipo E. nudoso y E. polimorfo, quiero resumir los eignos clínicos de estos episodios agudos, aprovechando algunos apuntes que, sabre "R. L." "Brote agudo" o "fiebre leprosa", recopiló el autor en el Lazareto de Agua de Dios Y que no han sido puhlicados (4. La reaccion leprosa (R. L. un episodic agudo que se observa eb ocasiones como primera manifestación de la lepra o durante su evolución crónica. Está caracterizada por fenomenos generales y síntomas localizados en la piel, en el sistema nervioso periferico o en ambas partes. Al mismo tiempo que las manifestaciones existentes de la enfermedad en la piel, los nervios y algunas víscerae se agudizan, se observan efluorescencias cutáneas nuevas de evolución rápida, aoompañadas de manifestaciones generales, independientes del cuadro clínico de la lepra. Es un episodio infeccioso que sobreviene con alguna frecuencia, en el trascurso de la dilatada evolución de la enfermedad.

  15. Investigation of a Medieval Pilgrim Burial Excavated from the Leprosarium of St Mary Magdalen Winchester, UK

    Science.gov (United States)

    Tucker, Katie; Filipek-Ogden, Kori; Montgomery, Janet; Cameron, Jamie; O’Connell, Tamsin; Evans, Jane; Marter, Phil; Taylor, G. Michael

    2017-01-01

    We have examined the remains of a Pilgrim burial from St Mary Magdalen, Winchester. The individual was a young adult male, aged around 18–25 years at the time of death. Radiocarbon dating showed the remains dated to the late 11th–early 12th centuries, a time when pilgrimages were at their height in Europe. Several lines of evidence in connection with the burial suggested this was an individual of some means and prestige. Although buried within the leprosarium cemetery, the skeleton showed only minimal skeletal evidence for leprosy, which was confined to the bones of the feet and legs. Nonetheless, molecular testing of several skeletal elements, including uninvolved bones all showed robust evidence of DNA from Mycobacterium leprae, consistent with the lepromatous or multibacillary form of the disease. We infer that in life, this individual almost certainly suffered with multiple soft tissue lesions. Genotyping of the M.leprae strain showed this belonged to the 2F lineage, today associated with cases from South-Central and Western Asia. During osteological examination it was noted that the cranium and facial features displayed atypical morphology for northern European populations. Subsequently, geochemical isotopic analyses carried out on tooth enamel indicated that this individual was indeed not local to the Winchester region, although it was not possible to be more specific about their geographic origin. PMID:28125649

  16. Immunological status of ENL (Erythema Nodosum Leprosum patients: its relationship to bacterial load and levels of circulanting IL-2R Perfil imunológico de pacientes com ENL (Erythema Nodosum Leprosum: relação entre carga bacilar e os níveis de IL-2R circulantes

    Directory of Open Access Journals (Sweden)

    Leila M.M. Vieira

    1996-04-01

    Full Text Available Recent data suggest that the clinical course of reactional states in leprosy is closely related to the cytokine profile released locally or systemically by the patients. In the present study, patients with erythema nodosum leprosum (ENL were grouped according to the intensity of their clinical symptoms. Clinical and immunological aspects of ENL and the impact of these parameters on bacterial load were assessed in conjunction with patients' in vitro immune response to mycobacterial antigens. In 10 out of the 17 patients tested, BI (bacterial index was reduced by at least 1 log from leprosy diagnosis to the onset of their first reactional episode (ENL, as compared to an expected 0.3 log reduction in the unreactional group for the same MDT (multidrug therapy period. However, no difference in the rate of BI reduction was noted at the end of MDT among ENL and unreactional lepromatous patients. Accordingly, although TNF-alpha (tumor necrosis factor levels were enhanced in the sera of 70.6% of the ENL patients tested, no relationship was noted between circulating TNF-alpha levels and the decrease in BI detected at the onset of the reactional episode. Evaluation of bacterial viability of M. leprae isolated from the reactional lesions showed no growth in the mouse footpads. Only 20% of the patients demonstrated specific immune response to M. leprae during ENL. Moreover, high levels of soluble IL-2R (interleukin-2 receptor were present in 78% of the patients. Circulating anti-neural (anti-ceramide and anti-galactocerebroside antibodies and anti-mycobacterial antibodies were detected in ENL patients' sera as well, which were not related to the clinical course of disease. Our data suggest that bacterial killing is enhanced during reactions. Emergence of specific immune response to M. leprae and the effective role of TNF-alpha in mediating fragmentation of bacteria still need to be clarified.Dados recentes sugerem que o curso clínico dos estados reacionais

  17. Genetic variants of the MAVS, MITA and MFN2 genes are not associated with leprosy in Han Chinese from Southwest China.

    Science.gov (United States)

    Wang, Dong; Li, Guo-Dong; Zhang, Deng-Feng; Xu, Ling; Li, Xiao-An; Yu, Xiu-Feng; Long, Heng; Li, Yu-Ye; Yao, Yong-Gang

    2016-11-01

    Leprosy is a chronic infectious disease caused by Mycobacterium leprae (M. leprae), which has massive genomic decay and dependence on host metabolism. Accumulating evidence showed a crucial role of mitochondria in metabolism and innate immunity. We hypothesized that the mitochondrial-related antimicrobial/antiviral immune genes MAVS (mitochondrial antiviral signaling protein), MITA (mediator of IRF3 activation) and MFN2 (mitofusin 2) would confer a risk to leprosy. In this study, we performed a case-control study to analyze 11 tag and/or non-synonymous SNPs of the MAVS, MITA and MFN2 genes in 527 leprosy patients and 583 healthy individuals, and directly sequenced the three genes in 80 leprosy patients with a family history from Yunnan, Southwest China. We found no association between these SNPs and leprosy (including its subtypes) based on the frequencies of alleles, genotypes and haplotypes between the cases and controls. There was also no enrichment of potential pathogenic variants of the three genes in leprosy patients. Our results suggested that genetic variants of the MAVS, MITA and MFN2 genes might not affect the susceptibility to leprosy. Copyright © 2016 Elsevier B.V. All rights reserved.

  18. Electrophysiological study of nerves in type-II reaction in leprosy

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    Santosh Kumar Singh

    2017-01-01

    Full Text Available Background: Leprosy is a chronic granulomatous infection primarily affecting the peripheral nervous system, skin and reticuloendothelial system. Cutaneous nerves are severely affected in lepra reaction and this leads to morbidity. Objective: To study electrophysiological pattern of different nerves involved in Type-II reactions in leprosy. Method: The present study was undertaken in 21 leprosy patients with Type-II reactions attending in and out-patient department of Dermatology & Venereology, B.R.D. Medical College, Gorakhpur from July 2005 to October 2006. This was a prospective case control study in which 20 healthy, age and sex matched people with no evidence of any disease (particularly nerve involvement were included. Limitation: Lesser number of cases were studied. Result: The proximal motor conduction latency was significantly prolonged in both ulnar and common peroneal nerve and proximal motor conduction velocity was also significantly reduced. On examining the values beyond 2S.D. of the control value, distal latency was not affected and only proximal conduction was affected in ulnar nerve. Conclusion: In Type II lepra reaction the motor conduction abnormalities are not prominent. Abnormalities are relatively more marked in the proximal segment.

  19. Some aspects of the pathogenesis and pathomorphology of leprosy*

    Science.gov (United States)

    Ivanova, N. A.

    1972-01-01

    The results of bacteriological and histological examinations of the organs of mice infected in the footpad with human leprosy bacilli by the Shepard method are summarized. The periods of observation ranged from a few days to 2 years. The disease developed in mice as a chronic infection with a lengthy “incubation” period of 3-4 months or more without symptoms. Usually about 2 years after inoculation, or in some cases after a shorter interval, the disease became generalized. Different types of tissue reaction occurred: simple inflammatory infiltration, tuberculoid granuloma, and leproma-like granuloma, similar to the clinical types of leprosy in man. When the infection became general, cells resembling lepra cells formed in the internal organs and at the site of infection. These cells contained massive intracellular aggregations of mycobacteria and compact globi with lipids and a vacuolized protoplasm, similar in origin and morphology to the lepra cells in human lesions. Changes were found in the neuroreceptor apparatus of the skin of the mice, with groups of leprosy bacilli in the endoneurium, the Schwann cells, and the perineural spaces. The length of the experiments and the histological examination of material from the sacrificed mice at different stages of the infective process revealed biological parallels with human leprosy and threw light on a number of aspects of the pathogenesis of leprosy. ImagesFig. 2Fig. 3Fig. 4Fig. 5Fig. 1 A-DFig. 10Fig. 11Fig. 12Fig. 13Fig. 6Fig. 7Fig. 8Fig. 9 PMID:4538195

  20. Leprosy - one of the many forgotten tropical diseases.

    Science.gov (United States)

    Zwolska, Zofia; Augustynowicz-Kopeć, Ewa

    2017-02-14

    Leprosy or Hansen disease is caused by an infection of Mycobacterium leprae. The large number of undetected cases (2000-2012 years 4 mln people) remains a threat to the elimination of leprosy. Leprosy is an unheard in Poland and generally is considered a condition so "exotic" that it is not worth to spend more attention to it. Forgotten disease in developed countries still thrives in an environment of poor and uneducated. Regardless of the conclusion that in the 21st century none infectious disease should not be treated as a disease on the designated regions of the world, other than our own, it should be recalled that the M. leprae was discovered in Europe, where for many years there were leprosaria and still infectious hospitals in Great Brittan, France or Spain get patients suspected of leprosy. The mobility of the inhabitants of the globe caused by wars, ethnic conflicts or a simple tourism causes that any infectious disease can not be treated as solely limited to distant us regions. The best proof of this were the viral diseases, formerly found in only in Asia or Africa, and currently transmitted to Europe [1]. At any moment, we can stand up against the problem of diagnostics of humans toward leprosy. Many medical reports indicate that leprosy as a disease with many symptoms encountered difficulties in its diagnosis. Only the experience of medical professionals and good microbiological diagnosis may speed up the diagnosis of leprosy.