Mycobacterium ulcerans infection (Buruli ulcer) is a neglected but treatable skin disease endemic in over
30 countries. M. ulcerans is an environmental mycobacteria with an elusive mode of transmission to humans. Ecological and Molecular epidemiological studies to identify reservoirs and transmission vectors are important
for source tracking infections especially during outbreaks and elucidating transmission routes.
Potentials of 3D models in anticancer drug screeningAnjali R.
A short presentation about the differences between 2D and 3D culture models, why researchers are moving toward 3D models in anticancer drug screening, the methods used in doing so and a recent case study of 3D tumour model being used for drug screening.
El lunes 23 de octubre de 2017 celebramos una jornada en la Fundación Ramón Areces sobre Microbiota Intestinal: Implicaciones en la Salud y Enfermedad.
Dr. Talita Resende - Organoids as an invitro model for enteric diseasesJohn Blue
1) The document discusses using intestinal organoids as an in vitro model to study enteric diseases in pigs. Organoids mimic intestinal tissue and present all organ-specific cell types.
2) The author's project uses mouse enteroids to study the pathogen Lawsonia intracellularis, which causes proliferative enteropathy in pigs. Preliminary results show the enteroids are permissive to L. intracellularis infection.
3) The goal is to further study the mechanisms of proliferation caused by L. intracellularis using the enteroid model to develop new treatment and control strategies for the disease. The author is also working to characterize and develop pig intestinal organoids for host-pathogen interaction studies.
This document is a curriculum vitae for Priyanka Voori Giri. It provides her contact information, educational background including an M.Tech in International Biotechnology, and research experience working on projects related to cancer stem cells, anti-inflammatory effects, and bacteriostatic/bactericidal nanoparticles. It also lists her publications, awards, skills in molecular biology, cell culture, and bioinformatics techniques, and areas of interest in inflammation, immunology and cancer biology.
The Evolution of In Situ Genetic Technologyasclepiuspdfs
In situ genetic technology was historically developed and mainly focused on detection purpose, allowing specific nucleic acid sequences to be visualized in morphologically preserved tissue sections. With the synergy of genetics and immunohistochemistry, in situ detection can correlate microscopic topological information with gene activity at the transcriptional or post-transcriptional levels in specific tissues. Furthermore, its resolution allows spatial distribution of nucleic acid products to be revealed in a heterogeneous cell population. The newest member to the franchise of in situ genetic technology is a direct-on-specimen enrichment methodology specifically for cell-free DNA liquid biopsy. Contrary to in situ detection, this in-well in situ innovation tackles the very first sample preparation step to reduce material loss, thereby improving overall sensitivity. Genomic nucleic acids purified from specimens have been proven to be time consuming and suffered from damages and losses; the evolution of in situ genetic technology offers a powerful tool for precision functional genomics, enabling cross-check between in vitro and in vivo findings. It further opens the door to ultimate genetic engineering in situ.
The culture of cells in two dimensions does not reproduce the histological characteristics of a tissue for informative or useful study. Growing cells as three-dimensional (3D) models more analogous to their existence in vivo may be more clinically relevant. Discuss the potential of using three dimensional cell cultures for anti-cancer drug screening.
Genes and Tissue Culture Assignment Presentation (Group 3)Lim Ke Wen
The culture of cells in two dimensions does not reproduce the histological characteristics of a tissue for informative or useful study. Growing cells as three-dimensional (3D) models more analogous to their existence in vivo may be more clinically relevant. Discuss the potential of using three dimensional cell cultures for anti-cancer drug screening.
This document describes the development of a fluorescence-based assay called "ProteAl" to detect the volatile biomarker 2-methylbutanal produced by Proteus bacteria. Gas chromatography-mass spectrometry and Fourier transform infrared spectroscopy were used to identify 2-methylbutanal in the headspace of Proteus cultures. A fluorescent dye, 5-dimethylaminonaphthalene-1-sulfonylhydrazine, was found to react specifically with 2-methylbutanal, producing a distinct green fluorescence. Testing of 95 bacterial strains showed the ProteAl assay can identify Proteus with 100% specificity and sensitivity, providing a simple method for rapid surveillance of this pathogen.
Potentials of 3D models in anticancer drug screeningAnjali R.
A short presentation about the differences between 2D and 3D culture models, why researchers are moving toward 3D models in anticancer drug screening, the methods used in doing so and a recent case study of 3D tumour model being used for drug screening.
El lunes 23 de octubre de 2017 celebramos una jornada en la Fundación Ramón Areces sobre Microbiota Intestinal: Implicaciones en la Salud y Enfermedad.
Dr. Talita Resende - Organoids as an invitro model for enteric diseasesJohn Blue
1) The document discusses using intestinal organoids as an in vitro model to study enteric diseases in pigs. Organoids mimic intestinal tissue and present all organ-specific cell types.
2) The author's project uses mouse enteroids to study the pathogen Lawsonia intracellularis, which causes proliferative enteropathy in pigs. Preliminary results show the enteroids are permissive to L. intracellularis infection.
3) The goal is to further study the mechanisms of proliferation caused by L. intracellularis using the enteroid model to develop new treatment and control strategies for the disease. The author is also working to characterize and develop pig intestinal organoids for host-pathogen interaction studies.
This document is a curriculum vitae for Priyanka Voori Giri. It provides her contact information, educational background including an M.Tech in International Biotechnology, and research experience working on projects related to cancer stem cells, anti-inflammatory effects, and bacteriostatic/bactericidal nanoparticles. It also lists her publications, awards, skills in molecular biology, cell culture, and bioinformatics techniques, and areas of interest in inflammation, immunology and cancer biology.
The Evolution of In Situ Genetic Technologyasclepiuspdfs
In situ genetic technology was historically developed and mainly focused on detection purpose, allowing specific nucleic acid sequences to be visualized in morphologically preserved tissue sections. With the synergy of genetics and immunohistochemistry, in situ detection can correlate microscopic topological information with gene activity at the transcriptional or post-transcriptional levels in specific tissues. Furthermore, its resolution allows spatial distribution of nucleic acid products to be revealed in a heterogeneous cell population. The newest member to the franchise of in situ genetic technology is a direct-on-specimen enrichment methodology specifically for cell-free DNA liquid biopsy. Contrary to in situ detection, this in-well in situ innovation tackles the very first sample preparation step to reduce material loss, thereby improving overall sensitivity. Genomic nucleic acids purified from specimens have been proven to be time consuming and suffered from damages and losses; the evolution of in situ genetic technology offers a powerful tool for precision functional genomics, enabling cross-check between in vitro and in vivo findings. It further opens the door to ultimate genetic engineering in situ.
The culture of cells in two dimensions does not reproduce the histological characteristics of a tissue for informative or useful study. Growing cells as three-dimensional (3D) models more analogous to their existence in vivo may be more clinically relevant. Discuss the potential of using three dimensional cell cultures for anti-cancer drug screening.
Genes and Tissue Culture Assignment Presentation (Group 3)Lim Ke Wen
The culture of cells in two dimensions does not reproduce the histological characteristics of a tissue for informative or useful study. Growing cells as three-dimensional (3D) models more analogous to their existence in vivo may be more clinically relevant. Discuss the potential of using three dimensional cell cultures for anti-cancer drug screening.
This document describes the development of a fluorescence-based assay called "ProteAl" to detect the volatile biomarker 2-methylbutanal produced by Proteus bacteria. Gas chromatography-mass spectrometry and Fourier transform infrared spectroscopy were used to identify 2-methylbutanal in the headspace of Proteus cultures. A fluorescent dye, 5-dimethylaminonaphthalene-1-sulfonylhydrazine, was found to react specifically with 2-methylbutanal, producing a distinct green fluorescence. Testing of 95 bacterial strains showed the ProteAl assay can identify Proteus with 100% specificity and sensitivity, providing a simple method for rapid surveillance of this pathogen.
Use of Simulation- based Training for Cancer Education among Nigerian Cliniciansasclepiuspdfs
Background: Among the many limitations of cancer control in Nigeria are lower awareness/competence and poorer training of health-care professionals (HCP). These manifest as deficiencies in advocacy, screening/diagnostic practices, and patient management. Medical simulation (MS) using models is an effective approach for sustainably improving the competence of HCP, especially regarding clinical breast examination (CBE), pelvic examination (PE), and digital rectal examination (DRE). The study evaluates the effect of MS during a Nigerian training course focusing on CBE, PE, and DRE. It answers the question: What is the immediate outcome of MS-based training, as well as the perspectives of HCP on the use of MS for cancer education? Methods: Participants included a convenience sample of Nigerian physicians and nurses who attended the American Society of Clinical Oncology-sponsored Multidisciplinary Cancer Management Course. The intervention was MS using high-fidelity models. The models demonstrated normal anatomic and common pathologic features of the breast, cervical, and prostate. Participants cycled through MS stations (i.e., CBE, PE, and DRE). Pre- and post-training surveys with comments evaluating self-reported comfort levels were the basis for comparison. Data analysis included descriptive statistics, Wilcoxon signed-rank test, Chi-square, and thematic analysis. Results: A total of 51 participants completed course evaluation forms (physicians - 35 and nurses - 16), with an average number of years in practice as 8 (±5.2) years. Pre-training survey showed non-significant differences in practices patterns; 71% (22/35) of physicians rarely performed PE (P=0.92), and 93% (14/16) of nurses rarely performed DRE (P=0.07). According to some participants, “the use of simulation is quite commendable as it gives room for improvement before using a human; it is the best method of learning I have ever enjoyed.” Conclusion: MS-based training significantly improved the comfort levels of participants regarding CBE and PE, as well as their likelihood to perform CBE, PE, and DRE. Participants recommend widespread use of MS for continuing medical education and undergraduate training.
Microbial source tracking markers for detection of fecal contamination in env...Asima Zehra
This presentation is an overview of the microbial source tracking markers that will be helpful for the beginners to understand its importance and challenges.
This curriculum vitae outlines the educational and professional background of Dr. Md. Anowar Khasru Parvez, a professor of microbiology. It details his educational qualifications including multiple PhD and MSc degrees. It also lists his employment history including positions at Jahangirnagar University and the Islamic University in Bangladesh. Finally, it provides an overview of his research expertise, projects, publications, and laboratory techniques.
1) Microbial identification determines the broad or narrow group to which a microorganism belongs. It is important for characterizing microorganisms detected in pharmaceutical products and facilities.
2) The U.S. Pharmacopeia outlines requirements for microbial identification in various chapters related to nonsterile products, sterility testing, and environmental monitoring. Identification to the species or strain level can provide insights for assessing and mitigating risks.
3) Identification methods include phenotypic methods based on biochemical reactions and genotypic methods based on nucleic acid sequences. Phenotypic methods are commonly used while genotypic methods are more reliable but require more specialized equipment and are typically used for critical investigations. The "polyphasic taxonomy" approach uses multiple
This curriculum vitae summarizes the educational and professional experience of Mirta Milić. She received her PhD in 2010 from the Faculty of Science in Zagreb, Croatia. Since 2011, she has worked as a Scientific Associate at the Institute for Medical Research and Occupational Health in Zagreb, focusing on genotoxicology and genetic monitoring. She also spent over a year as a postdoctoral fellow at the IRCCS San Raffaele Pisana in Rome, Italy from 2013-2014. Her research interests include genetic toxicology, biomarkers of exposure to various agents, and genetic monitoring. She has authored or co-authored several peer-reviewed publications in her field.
Development of pancreatic cancer organoid model for studying immune response ...TÀI LIỆU NGÀNH MAY
Để xem full tài liệu Xin vui long liên hệ page để được hỗ trợ
: https://www.facebook.com/thuvienluanvan01
HOẶC
https://www.facebook.com/garmentspace/
https://www.facebook.com/thuvienluanvan01
https://www.facebook.com/thuvienluanvan01
tai lieu tong hop, thu vien luan van, luan van tong hop, do an chuyen nganh
1) The document discusses the history and definitions of biomarkers from the 1960s to 1990s based on a literature review. It shows a progression from early uses related to diagnosing cancer to later applications in toxicology.
2) By the 1980s, biomarkers were being used to study occupational exposure and effects of toxic substances. Studies also began exploring their potential role in risk assessment.
3) In the 1990s, biomarkers were applied to monitoring the effects of interventions aimed at reducing disease risk by modifying exposure to suspected causal agents. This supported establishing exposure-disease relationships.
The document discusses how technology has helped shed light on cancer through research using large facilities like synchrotron radiation and neutron laboratories. Over 100,000 protein structures have been determined using these techniques to better understand biochemical processes and design drugs. Countries are investing in new facilities to advance scientific development and tackle challenges like cancer. Nanotechnology and drug delivery systems combined with characterization techniques can improve cancer treatment methods.
Simon Cooper received his Ph.D. in Radiation/Oncology and has since focused his research career on translating scientific findings into clinical applications while emphasizing patient care. He has authored multiple peer-reviewed articles in collaboration with clinicians and currently leads pre-clinical studies at Mayo Clinic Florida identifying potential new cancer therapies. Cooper aims to continue interacting with clinical researchers to advance innovative patient care.
This document summarizes a presentation on applying systems biology approaches to epidemiology and personalized medicine. It discusses how epidemiology has evolved from studying single risk factors to investigating complex interactions between genes, environments and diseases. Systems approaches integrate multiple levels of data to gain insights into disease mechanisms. Large consortia now apply these methods to conditions like asthma and COPD to understand their complexity. Epidemiology contributes a population perspective and helps develop evidence-based systems medicine frameworks.
The document provides information about postgraduate studies and PhD projects available at the Garvan Institute of Medical Research. It begins with an introduction to postgraduate studies at Garvan and its partnership with UNSW. It then describes the research divisions and groups at Garvan conducting research in cancer, immunological diseases, diabetes, metabolic diseases, neurological diseases, bone diseases, and bioinformatics. Specific PhD project descriptions are provided for several research groups in areas like cancer cell invasion, pancreatic carcinogenesis, B cell biology, immunology and more. Contact information is provided for supervisors of the different projects.
CovidOnTheWeb : covid19 linked data published on the WebFabien Gandon
The Covid-on-the-Web project aims to allow biomedical researchers to access, query and make sense of COVID-19 related literature. To do so, it adapts, combines and extends tools to process, analyze and enrich the "COVID-19 Open Research Dataset" (CORD-19) that gathers 50,000+ full-text scientific articles related to the coronaviruses. We report on the RDF dataset and software resources produced in this project by leveraging skills in knowledge representation, text, data and argument mining, as well as data visualization and exploration. The dataset comprises two main knowledge graphs describing (1) named entities mentioned in the CORD-19 corpus and linked to DBpedia, Wikidata and other BioPortal vocabularies, and (2) arguments extracted using ACTA, a tool automating the extraction and visualization of argumentative graphs, meant to help clinicians analyze clinical trials and make decisions. On top of this dataset, we provide several visualization and exploration tools based on the Corese Semantic Web platform, MGExplorer visualization library, as well as the Jupyter Notebook technology. All along this initiative, we have been engaged in discussions with healthcare and medical research institutes to align our approach with the actual needs of the biomedical community, and we have paid particular attention to comply with the open and reproducible science goals, and the FAIR principles.
This document discusses changes in the landscape of cholangiocarcinoma. Molecular diagnostics using next generation sequencing can identify actionable mutations in 70-78% of intrahepatic cholangiocarcinoma cases, including IDH1, FGFR2, and BRAF mutations. Targeted therapies are available for many of these mutations. Immunotherapy may also benefit cholangiocarcinoma given its association with immune checkpoint proteins. Cell-free DNA analysis shows promise for early diagnosis and monitoring. Liver transplantation is being explored for selected intrahepatic cholangiocarcinoma cases treated with neoadjuvant therapies, with 3-year survival rates of 83% reported in one study.
Vasudevan Ayyappan is seeking new job opportunities and has over 10 years of experience in molecular biological techniques and 5+ years experience in next generation sequencing, genome editing, and pathway analysis. He has a broad range of skills in mammalian, microbial, and plant research and has effectively collaborated on multiple research projects.
This document discusses the polymicrobial nature of biofilm infections. It summarizes that biofilms are usually polymicrobial communities that provide advantages like passive resistance, metabolic cooperation, and an enlarged gene pool through horizontal gene transfer. DNA methods using PCR and sequencing have demonstrated the ability to identify microorganisms in clinical biofilm infections. A robust model of polymicrobial biofilm infection along with accurate diagnosis is improving clinical outcomes.
Natural fluorescence of normal and neoplastic human colonguest2f80ca
This document describes a study that used microspectrofluorometry to analyze and compare the natural fluorescence of normal and neoplastic (cancerous) human colon tissue. The study found:
1) Normal and cancerous colon tissues exhibited different patterns of fluorescence intensity and spectral shape, related to their distinct histological organizations.
2) The most evident spectral differences involved the stromal compartment and were likely due to different fluorochromes, related to the host response to tumors.
3) The nature and extent of autofluorescence modifications between normal and cancerous tissues helped explain previous "in vivo" analysis findings and highlighted the importance of excitation parameters for exploiting autofluorescence in diagnosis.
MALDI-TOF MS is an emerging technique for microbial identification, characterization, and typing that provides protein fingerprints unique to each microorganism. This review discusses applications of MALDI-TOF MS in environmental microbiology, including its use for identifying environmental microorganisms, bacterial strain typing, and research in bioremediation. Some parameters that can influence the reproducibility of MALDI-TOF MS results are also discussed.
This document summarizes the results of sequencing and analyzing genomes of Mycobacterium tuberculosis isolates from 5 patients with extra-pulmonary tuberculosis. Key findings include:
1. The isolates showed genetic heterogeneity, with variations in single nucleotide polymorphisms and presence/absence of genes compared to the reference genome.
2. One isolate (LN8) showed the highest number of unique single nucleotide variations and gene deletions, indicating it had diverged more than the others.
3. Several genes missing or disrupted in the isolates are involved in important processes like cell wall biosynthesis and membrane transport, which may influence pathogenesis.
4. The variations identified suggest next-generation sequencing can effectively detect small genomic changes in M
International Journal of Engineering Research and Applications (IJERA) is an open access online peer reviewed international journal that publishes research and review articles in the fields of Computer Science, Neural Networks, Electrical Engineering, Software Engineering, Information Technology, Mechanical Engineering, Chemical Engineering, Plastic Engineering, Food Technology, Textile Engineering, Nano Technology & science, Power Electronics, Electronics & Communication Engineering, Computational mathematics, Image processing, Civil Engineering, Structural Engineering, Environmental Engineering, VLSI Testing & Low Power VLSI Design etc.
This document is a resume for Ho Wing Sze, a molecular microbiologist seeking a position in a highly-caliber company. Ho Wing Sze has a PhD in Molecular Microbiology from the University of Malaya and investigated antimicrobial resistance mechanisms and genomic diversity of E. coli in Malaysia. Ho has extensive research experience investigating foodborne pathogens such as Salmonella and E. coli using techniques including next-generation sequencing and phenotype microarray analysis. Ho Wing Sze has authored several publications and presented research at international conferences.
Biological contamination is the dread of every person working with cell culture. When cultures become infected with microorganisms, or cross-contaminated by foreign cells, these cultures usually must be destroyed. Since the sources of culture contamination are ubiquitous as well as difficult to identify and eliminate, no cell culture laboratory remains unaffected by this concern. With the continuing increase in the use of cell culture for biological research, vaccine production, and production of therapeutic proteins for personalized medicine and emerging regenerative medicine applications, culture contamination remains a highly important issue. Cell line cross-contamination can be a problem for scientists working with cultured cells. Studies suggest anywhere from 15–20% of the time, cells used in experiments have been misidentified or contaminated with another cell line. Problems with cell line cross-contamination have even been detected in lines from the NCI-60 panel, which are used routinely for drug-screening studies. Major cell line repositories, including the American Type Culture Collection (ATCC), the European Collection of Cell Cultures (ECACC) and the German Collection of Microorganisms and Cell Cultures (DSMZ), have received cell line submissions from researchers that were misidentified by them. Such contamination poses a problem for the quality of research produced using cell culture lines, and the major repositories are now authenticating all cell line submissions. ATCC uses short tandem repeat (STR) DNA fingerprinting to authenticate its cell lines.
potassium, chloride, bicarbonate, blood urea nitrogen (BUN), magnesium, creatinine, glucose, and sometimes calcium. Tests that focus on cholesterol levels can determine LDL and HDL cholesterol levels, as well as triglyceride levels.[6]
Use of Simulation- based Training for Cancer Education among Nigerian Cliniciansasclepiuspdfs
Background: Among the many limitations of cancer control in Nigeria are lower awareness/competence and poorer training of health-care professionals (HCP). These manifest as deficiencies in advocacy, screening/diagnostic practices, and patient management. Medical simulation (MS) using models is an effective approach for sustainably improving the competence of HCP, especially regarding clinical breast examination (CBE), pelvic examination (PE), and digital rectal examination (DRE). The study evaluates the effect of MS during a Nigerian training course focusing on CBE, PE, and DRE. It answers the question: What is the immediate outcome of MS-based training, as well as the perspectives of HCP on the use of MS for cancer education? Methods: Participants included a convenience sample of Nigerian physicians and nurses who attended the American Society of Clinical Oncology-sponsored Multidisciplinary Cancer Management Course. The intervention was MS using high-fidelity models. The models demonstrated normal anatomic and common pathologic features of the breast, cervical, and prostate. Participants cycled through MS stations (i.e., CBE, PE, and DRE). Pre- and post-training surveys with comments evaluating self-reported comfort levels were the basis for comparison. Data analysis included descriptive statistics, Wilcoxon signed-rank test, Chi-square, and thematic analysis. Results: A total of 51 participants completed course evaluation forms (physicians - 35 and nurses - 16), with an average number of years in practice as 8 (±5.2) years. Pre-training survey showed non-significant differences in practices patterns; 71% (22/35) of physicians rarely performed PE (P=0.92), and 93% (14/16) of nurses rarely performed DRE (P=0.07). According to some participants, “the use of simulation is quite commendable as it gives room for improvement before using a human; it is the best method of learning I have ever enjoyed.” Conclusion: MS-based training significantly improved the comfort levels of participants regarding CBE and PE, as well as their likelihood to perform CBE, PE, and DRE. Participants recommend widespread use of MS for continuing medical education and undergraduate training.
Microbial source tracking markers for detection of fecal contamination in env...Asima Zehra
This presentation is an overview of the microbial source tracking markers that will be helpful for the beginners to understand its importance and challenges.
This curriculum vitae outlines the educational and professional background of Dr. Md. Anowar Khasru Parvez, a professor of microbiology. It details his educational qualifications including multiple PhD and MSc degrees. It also lists his employment history including positions at Jahangirnagar University and the Islamic University in Bangladesh. Finally, it provides an overview of his research expertise, projects, publications, and laboratory techniques.
1) Microbial identification determines the broad or narrow group to which a microorganism belongs. It is important for characterizing microorganisms detected in pharmaceutical products and facilities.
2) The U.S. Pharmacopeia outlines requirements for microbial identification in various chapters related to nonsterile products, sterility testing, and environmental monitoring. Identification to the species or strain level can provide insights for assessing and mitigating risks.
3) Identification methods include phenotypic methods based on biochemical reactions and genotypic methods based on nucleic acid sequences. Phenotypic methods are commonly used while genotypic methods are more reliable but require more specialized equipment and are typically used for critical investigations. The "polyphasic taxonomy" approach uses multiple
This curriculum vitae summarizes the educational and professional experience of Mirta Milić. She received her PhD in 2010 from the Faculty of Science in Zagreb, Croatia. Since 2011, she has worked as a Scientific Associate at the Institute for Medical Research and Occupational Health in Zagreb, focusing on genotoxicology and genetic monitoring. She also spent over a year as a postdoctoral fellow at the IRCCS San Raffaele Pisana in Rome, Italy from 2013-2014. Her research interests include genetic toxicology, biomarkers of exposure to various agents, and genetic monitoring. She has authored or co-authored several peer-reviewed publications in her field.
Development of pancreatic cancer organoid model for studying immune response ...TÀI LIỆU NGÀNH MAY
Để xem full tài liệu Xin vui long liên hệ page để được hỗ trợ
: https://www.facebook.com/thuvienluanvan01
HOẶC
https://www.facebook.com/garmentspace/
https://www.facebook.com/thuvienluanvan01
https://www.facebook.com/thuvienluanvan01
tai lieu tong hop, thu vien luan van, luan van tong hop, do an chuyen nganh
1) The document discusses the history and definitions of biomarkers from the 1960s to 1990s based on a literature review. It shows a progression from early uses related to diagnosing cancer to later applications in toxicology.
2) By the 1980s, biomarkers were being used to study occupational exposure and effects of toxic substances. Studies also began exploring their potential role in risk assessment.
3) In the 1990s, biomarkers were applied to monitoring the effects of interventions aimed at reducing disease risk by modifying exposure to suspected causal agents. This supported establishing exposure-disease relationships.
The document discusses how technology has helped shed light on cancer through research using large facilities like synchrotron radiation and neutron laboratories. Over 100,000 protein structures have been determined using these techniques to better understand biochemical processes and design drugs. Countries are investing in new facilities to advance scientific development and tackle challenges like cancer. Nanotechnology and drug delivery systems combined with characterization techniques can improve cancer treatment methods.
Simon Cooper received his Ph.D. in Radiation/Oncology and has since focused his research career on translating scientific findings into clinical applications while emphasizing patient care. He has authored multiple peer-reviewed articles in collaboration with clinicians and currently leads pre-clinical studies at Mayo Clinic Florida identifying potential new cancer therapies. Cooper aims to continue interacting with clinical researchers to advance innovative patient care.
This document summarizes a presentation on applying systems biology approaches to epidemiology and personalized medicine. It discusses how epidemiology has evolved from studying single risk factors to investigating complex interactions between genes, environments and diseases. Systems approaches integrate multiple levels of data to gain insights into disease mechanisms. Large consortia now apply these methods to conditions like asthma and COPD to understand their complexity. Epidemiology contributes a population perspective and helps develop evidence-based systems medicine frameworks.
The document provides information about postgraduate studies and PhD projects available at the Garvan Institute of Medical Research. It begins with an introduction to postgraduate studies at Garvan and its partnership with UNSW. It then describes the research divisions and groups at Garvan conducting research in cancer, immunological diseases, diabetes, metabolic diseases, neurological diseases, bone diseases, and bioinformatics. Specific PhD project descriptions are provided for several research groups in areas like cancer cell invasion, pancreatic carcinogenesis, B cell biology, immunology and more. Contact information is provided for supervisors of the different projects.
CovidOnTheWeb : covid19 linked data published on the WebFabien Gandon
The Covid-on-the-Web project aims to allow biomedical researchers to access, query and make sense of COVID-19 related literature. To do so, it adapts, combines and extends tools to process, analyze and enrich the "COVID-19 Open Research Dataset" (CORD-19) that gathers 50,000+ full-text scientific articles related to the coronaviruses. We report on the RDF dataset and software resources produced in this project by leveraging skills in knowledge representation, text, data and argument mining, as well as data visualization and exploration. The dataset comprises two main knowledge graphs describing (1) named entities mentioned in the CORD-19 corpus and linked to DBpedia, Wikidata and other BioPortal vocabularies, and (2) arguments extracted using ACTA, a tool automating the extraction and visualization of argumentative graphs, meant to help clinicians analyze clinical trials and make decisions. On top of this dataset, we provide several visualization and exploration tools based on the Corese Semantic Web platform, MGExplorer visualization library, as well as the Jupyter Notebook technology. All along this initiative, we have been engaged in discussions with healthcare and medical research institutes to align our approach with the actual needs of the biomedical community, and we have paid particular attention to comply with the open and reproducible science goals, and the FAIR principles.
This document discusses changes in the landscape of cholangiocarcinoma. Molecular diagnostics using next generation sequencing can identify actionable mutations in 70-78% of intrahepatic cholangiocarcinoma cases, including IDH1, FGFR2, and BRAF mutations. Targeted therapies are available for many of these mutations. Immunotherapy may also benefit cholangiocarcinoma given its association with immune checkpoint proteins. Cell-free DNA analysis shows promise for early diagnosis and monitoring. Liver transplantation is being explored for selected intrahepatic cholangiocarcinoma cases treated with neoadjuvant therapies, with 3-year survival rates of 83% reported in one study.
Vasudevan Ayyappan is seeking new job opportunities and has over 10 years of experience in molecular biological techniques and 5+ years experience in next generation sequencing, genome editing, and pathway analysis. He has a broad range of skills in mammalian, microbial, and plant research and has effectively collaborated on multiple research projects.
This document discusses the polymicrobial nature of biofilm infections. It summarizes that biofilms are usually polymicrobial communities that provide advantages like passive resistance, metabolic cooperation, and an enlarged gene pool through horizontal gene transfer. DNA methods using PCR and sequencing have demonstrated the ability to identify microorganisms in clinical biofilm infections. A robust model of polymicrobial biofilm infection along with accurate diagnosis is improving clinical outcomes.
Natural fluorescence of normal and neoplastic human colonguest2f80ca
This document describes a study that used microspectrofluorometry to analyze and compare the natural fluorescence of normal and neoplastic (cancerous) human colon tissue. The study found:
1) Normal and cancerous colon tissues exhibited different patterns of fluorescence intensity and spectral shape, related to their distinct histological organizations.
2) The most evident spectral differences involved the stromal compartment and were likely due to different fluorochromes, related to the host response to tumors.
3) The nature and extent of autofluorescence modifications between normal and cancerous tissues helped explain previous "in vivo" analysis findings and highlighted the importance of excitation parameters for exploiting autofluorescence in diagnosis.
MALDI-TOF MS is an emerging technique for microbial identification, characterization, and typing that provides protein fingerprints unique to each microorganism. This review discusses applications of MALDI-TOF MS in environmental microbiology, including its use for identifying environmental microorganisms, bacterial strain typing, and research in bioremediation. Some parameters that can influence the reproducibility of MALDI-TOF MS results are also discussed.
This document summarizes the results of sequencing and analyzing genomes of Mycobacterium tuberculosis isolates from 5 patients with extra-pulmonary tuberculosis. Key findings include:
1. The isolates showed genetic heterogeneity, with variations in single nucleotide polymorphisms and presence/absence of genes compared to the reference genome.
2. One isolate (LN8) showed the highest number of unique single nucleotide variations and gene deletions, indicating it had diverged more than the others.
3. Several genes missing or disrupted in the isolates are involved in important processes like cell wall biosynthesis and membrane transport, which may influence pathogenesis.
4. The variations identified suggest next-generation sequencing can effectively detect small genomic changes in M
International Journal of Engineering Research and Applications (IJERA) is an open access online peer reviewed international journal that publishes research and review articles in the fields of Computer Science, Neural Networks, Electrical Engineering, Software Engineering, Information Technology, Mechanical Engineering, Chemical Engineering, Plastic Engineering, Food Technology, Textile Engineering, Nano Technology & science, Power Electronics, Electronics & Communication Engineering, Computational mathematics, Image processing, Civil Engineering, Structural Engineering, Environmental Engineering, VLSI Testing & Low Power VLSI Design etc.
This document is a resume for Ho Wing Sze, a molecular microbiologist seeking a position in a highly-caliber company. Ho Wing Sze has a PhD in Molecular Microbiology from the University of Malaya and investigated antimicrobial resistance mechanisms and genomic diversity of E. coli in Malaysia. Ho has extensive research experience investigating foodborne pathogens such as Salmonella and E. coli using techniques including next-generation sequencing and phenotype microarray analysis. Ho Wing Sze has authored several publications and presented research at international conferences.
Biological contamination is the dread of every person working with cell culture. When cultures become infected with microorganisms, or cross-contaminated by foreign cells, these cultures usually must be destroyed. Since the sources of culture contamination are ubiquitous as well as difficult to identify and eliminate, no cell culture laboratory remains unaffected by this concern. With the continuing increase in the use of cell culture for biological research, vaccine production, and production of therapeutic proteins for personalized medicine and emerging regenerative medicine applications, culture contamination remains a highly important issue. Cell line cross-contamination can be a problem for scientists working with cultured cells. Studies suggest anywhere from 15–20% of the time, cells used in experiments have been misidentified or contaminated with another cell line. Problems with cell line cross-contamination have even been detected in lines from the NCI-60 panel, which are used routinely for drug-screening studies. Major cell line repositories, including the American Type Culture Collection (ATCC), the European Collection of Cell Cultures (ECACC) and the German Collection of Microorganisms and Cell Cultures (DSMZ), have received cell line submissions from researchers that were misidentified by them. Such contamination poses a problem for the quality of research produced using cell culture lines, and the major repositories are now authenticating all cell line submissions. ATCC uses short tandem repeat (STR) DNA fingerprinting to authenticate its cell lines.
potassium, chloride, bicarbonate, blood urea nitrogen (BUN), magnesium, creatinine, glucose, and sometimes calcium. Tests that focus on cholesterol levels can determine LDL and HDL cholesterol levels, as well as triglyceride levels.[6]
potassium, chloride, bicarbonate, blood urea nitrogen (BUN), magnesium, creatinine, glucose, and sometimes calcium. Tests that focus on cholesterol levels can determine LDL and HDL cholesterol levels, as well as triglyceride levels.[6]
potassium, chloride, bicarbonate, blood urea nitrogen (BUN), magnesium, creatinine, glucose, and sometimes calcium. Tests that focus on cholesterol levels can determine LDL and HDL cholesterol levels, as well as triglyceride levels.[6]
Prevalence and Antimicrobial Susceptibility of Methicillin Resistant StaphJoshua Owolabi
This document summarizes a study on the prevalence and antimicrobial susceptibility of methicillin resistant Staphylococcus aureus (MRSA) and coagulase-negative Staphylococci (CoNS) isolated from healthy university students in Nigeria. Swabs were collected from the noses and necks of 100 students. A total of 39 Staphylococcus species were identified, including MRSA and MRCoNS. The MRSA strains showed high resistance to methicillin and several other antibiotics. CoNS also demonstrated moderate to high resistance to several antibiotics tested. This highlights the need for surveillance of antibiotic resistance in the community and policies to prevent the spread of resistant infections.
International Journal of Biometrics and Bioinformatics(IJBB) Volume (3) Issue...CSCJournals
This document reviews approaches for predicting breast cancer prognosis using both clinical data and gene expression profiles. Traditional prognosis models rely mainly on clinical factors like age, tumor size, and lymph node status, but can fail to distinguish molecularly distinct subgroups. Gene expression profiling via microarray technology has improved molecular classification and shown promise for prognosis. However, most studies have focused on gene signatures without fully leveraging clinical data. Integrating clinical and gene expression data may enhance accuracy by accounting for their complementary nature. The review discusses feature selection and classification methods applied to both data types, as well as related work on data integration. The goal is to develop an improved prognosis model that incorporates both clinical and molecular factors.
Cancer remains one of the most formidable challenges in modern medicine, posing significant health burdens globally. In recent years, extensive research efforts have aimed at understanding the intricate mechanisms underlying cancer development, progression, and treatment. This review provides a comprehensive overview of the latest advancements in cancer research across various domains. The elucidation of genetic and molecular alterations driving oncogenesis has revolutionized our understanding of cancer biology. Key discoveries in genomics, transcriptomics, and proteomics have unveiled the heterogeneous nature of tumors, paving the way for personalized treatment approaches. Moreover, advancements in high-throughput sequencing technologies have facilitated the identification of novel cancer biomarkers with diagnostic, prognostic, and therapeutic implications. The tumor microenvironment (TME) has emerged as a critical determinant of cancer progression and therapy response. Research focusing on the dynamic interactions between cancer cells, immune cells, and stromal components within the TME has led to the development of immunotherapeutic strategies, including immune checkpoint inhibitors and adoptive cell therapies, which have demonstrated remarkable efficacy in various cancer types. In addition to targeted therapies and immunotherapies, the advent of precision medicine has transformed cancer treatment paradigms. Molecular profiling of tumors enables clinicians to match patients with specific targeted therapies, optimizing therapeutic outcomes while minimizing adverse effects. Furthermore, the integration of artificial intelligence and machine learning algorithms in cancer research has facilitated the prediction of treatment responses and identification of novel therapeutic targets.
Multiplex Detection of Antimicrobial Resistance Genes.pdfMaNoLo440315
This document describes a study that developed and tested a multiplex PCR assay to rapidly detect 24 common antibiotic resistance genes in urinary tract infection samples. The assay was tested on 366 bacterial isolates from a public antimicrobial resistance database which had been previously characterized through whole genome sequencing and phenotypic assays. Results from the multiplex PCR assay showed high accuracy in predicting bacterial identification and were mostly in agreement with previous genotypic and phenotypic characterization of antibiotic resistance. The study provides evidence that genetic diagnostic methods like multiplex PCR could enable more rapid identification of antibiotic resistance genes to help characterize and treat urinary tract infections.
Identification of bacteria and fungi in the solid waste generated in hospita...Dr. Gawad Alwabr
This study identified bacteria and fungi present in solid hospital waste in Sana'a, Yemen. Samples were collected from hospital wards, departments, and storage areas over 8 months. Testing identified the following microorganisms: Klebsiella spp., E. coli, Citrobacter spp., Candida spp., Proteus spp., Cladosporium werneckii spp., Bacillus spp., Aspergillus spp., Trichothecium spp., Mucor spp., and Acinetobacter spp. The types and amounts of microorganisms varied by season and location. This study confirms the presence of pathogenic bacteria and fungi in hospital solid waste that could spread infection
1) The study examined the cough aerosols of 128 patients colonized with MDROs like MRSA, ESBL, and VRE to see if the organisms could be transmitted onto culture plates.
2) In 18% of cases, viable MDROs were transmitted onto the culture plates through coughing.
3) Multivariate analysis showed that the strength of a patient's cough significantly predicted whether MDROs would be transmitted, with strong coughing making transmission over 7 times more likely.
4) The results suggest that the severity of a patient's cough should be considered when deciding whether single room isolation is needed for patients colonized with MDROs in their respiratory tract. However, more objective
1) The study measured viable microbial counts on culture plates after patients colonized with MDROs coughed. 18% of coughs resulted in MDRO growth on plates.
2) A multivariate analysis found that the strength of cough significantly predicted whether MDROs would grow on plates, with strong coughs being over 7 times more likely to result in growth.
3) The results suggest that the severity of a patient's cough should be considered when deciding whether single room isolation is needed for patients colonized with MDROs in their respiratory tract. However, more research is required to objectively assess cough severity.
Mohamad Al Kilani successfully completed an online non-credit course on Tropical Parasitology: Protozoans, Worms, Vectors and Human Diseases offered by Duke University on May 06, 2015. The course presented information on important human parasitic diseases, including their life cycles, transmission, distribution, pathophysiology, treatment, and prevention. It was taught by Professor John A. Bartlett of Duke University Medical Center along with Professors Franklin Mosha and Mramba Nyindo of Kilimanjaro Christian Medical University College in Tanzania.
This research article characterized the genetic diversity of Plasmodium vivax and P. falciparum populations from pregnant women in four malaria-endemic countries. Between 2008-2011, nearly 2000 pregnant women were recruited from Brazil, Colombia, India, and Papua New Guinea and followed until delivery, collecting blood samples. Seven P. vivax microsatellite markers were used to genotype 229 P. vivax isolates. P. vivax populations showed moderate to high genetic differentiation between countries and higher diversity than P. falciparum populations from the same areas. Diversity of P. vivax was very high in some settings compared to transmission levels, suggesting stable demographic histories.
IDENTIFYING MOSQUITO LARVAE AND ITS DISTRIBUTION.pptxGraceArmenio
The document discusses a study conducted at Aurora National Science High School to identify mosquito larvae species and their distribution across the school's grounds. Field surveys were carried out using simple random sampling to collect mosquito larvae specimens, which were then identified morphologically based on traits of the head, thorax, and abdomen. The study aims to understand the local mosquito population and assist in targeted mosquito control strategies. However, limitations include challenges in accurate species identification and seasonal variations in larvae distribution.
This document is the preface to the book "Salmonella: Methods and Protocols, Second Edition". It summarizes advances in Salmonella research since the first edition, including new methods for detecting and treating Salmonella infections, overcoming antibiotic resistance, vaccine development using nanotechnology, and using genetically modified Salmonella as delivery vectors for cancer therapies. The second edition features new chapters covering molecular detection and identification of Salmonella, quantitative proteomics to identify host factors in Salmonella infection, determining antibiotic resistance, and various microscopy and modeling methods. The preface expresses gratitude to the contributors for sharing their expertise and to the publisher.
The document discusses several use cases for applying data mining and machine learning techniques in healthcare and biomedical research. Three examples are:
1) Early diagnosis of cancers like lung cancer and breast cancer through predictive modeling of patient data to detect cancers at earlier stages when survival rates are higher.
2) Predicting patient responses to drug therapies for cancers like breast cancer by combining different types of molecular profiling data using techniques like support vector machines and random forests.
3) Using imaging data and temporal analysis of metrics like medication purchases to better understand and predict chronic diseases like diabetes and associated health complications.
Application of Biomedical Informatics in Clinical Problem Solvingimprovemed
1) The document discusses several use cases for applying data mining and machine learning techniques to biomedical problems.
2) One use case aims to enable early diagnosis of cancers like lung cancer through predictive modeling of patient data.
3) Another use case examines predicting patient responses to drug therapies for breast cancer by analyzing genomic and other molecular profiling data using machine learning algorithms.
4) Other use cases discussed include using imaging data and time series analysis of patient information to aid in early detection and risk assessment of chronic diseases.
DIAGNOSTICS - Diagnosis of TB - A Nanodiagnostic Approach.pdfsudeepbhattacharyya
The document discusses diagnosis of tuberculosis and highlights opportunities for nanotechnology-based diagnostic approaches. It summarizes several existing methods for TB diagnosis including microscopy, culture-based techniques, immunological methods, and molecular tests. However, current diagnostics have limitations such as low sensitivity, long turnaround time, and requirements for specialized equipment and facilities. The document proposes that nanodiagnostics utilizing nanoparticles, antigens, and antibodies may enable the development of improved point-of-care tests for more rapid, affordable and accurate TB detection.
1. The study developed a new PCR/RFLP technique to identify the 3 genotypes of Plasmodium vivax circumsporozoite protein (VK210, VK247, and P. vivax-like) using DNA extracted from blood samples.
2. The technique uses PCR amplification of the central immunodominant region of the CSP gene followed by restriction enzyme digestion and fragment analysis to distinguish the genotypes.
3. Testing demonstrated the technique could accurately identify the genotypes using plasmid controls for each variant, and that it had high sensitivity detecting parasitemia levels as low as 0.0069 parasites per microliter.
Know the difference between Endodontics and Orthodontics.Gokuldas Hospital
Your smile is beautiful.
Let’s be honest. Maintaining that beautiful smile is not an easy task. It is more than brushing and flossing. Sometimes, you might encounter dental issues that need special dental care. These issues can range anywhere from misalignment of the jaw to pain in the root of teeth.
5-hydroxytryptamine or 5-HT or Serotonin is a neurotransmitter that serves a range of roles in the human body. It is sometimes referred to as the happy chemical since it promotes overall well-being and happiness.
It is mostly found in the brain, intestines, and blood platelets.
5-HT is utilised to transport messages between nerve cells, is known to be involved in smooth muscle contraction, and adds to overall well-being and pleasure, among other benefits. 5-HT regulates the body's sleep-wake cycles and internal clock by acting as a precursor to melatonin.
It is hypothesised to regulate hunger, emotions, motor, cognitive, and autonomic processes.
NAVIGATING THE HORIZONS OF TIME LAPSE EMBRYO MONITORING.pdfRahul Sen
Time-lapse embryo monitoring is an advanced imaging technique used in IVF to continuously observe embryo development. It captures high-resolution images at regular intervals, allowing embryologists to select the most viable embryos for transfer based on detailed growth patterns. This technology enhances embryo selection, potentially increasing pregnancy success rates.
Promoting Wellbeing - Applied Social Psychology - Psychology SuperNotesPsychoTech Services
A proprietary approach developed by bringing together the best of learning theories from Psychology, design principles from the world of visualization, and pedagogical methods from over a decade of training experience, that enables you to: Learn better, faster!
These lecture slides, by Dr Sidra Arshad, offer a simplified look into the mechanisms involved in the regulation of respiration:
Learning objectives:
1. Describe the organisation of respiratory center
2. Describe the nervous control of inspiration and respiratory rhythm
3. Describe the functions of the dorsal and respiratory groups of neurons
4. Describe the influences of the Pneumotaxic and Apneustic centers
5. Explain the role of Hering-Breur inflation reflex in regulation of inspiration
6. Explain the role of central chemoreceptors in regulation of respiration
7. Explain the role of peripheral chemoreceptors in regulation of respiration
8. Explain the regulation of respiration during exercise
9. Integrate the respiratory regulatory mechanisms
10. Describe the Cheyne-Stokes breathing
Study Resources:
1. Chapter 42, Guyton and Hall Textbook of Medical Physiology, 14th edition
2. Chapter 36, Ganong’s Review of Medical Physiology, 26th edition
3. Chapter 13, Human Physiology by Lauralee Sherwood, 9th edition
- Video recording of this lecture in English language: https://youtu.be/Pt1nA32sdHQ
- Video recording of this lecture in Arabic language: https://youtu.be/uFdc9F0rlP0
- Link to download the book free: https://nephrotube.blogspot.com/p/nephrotube-nephrology-books.html
- Link to NephroTube website: www.NephroTube.com
- Link to NephroTube social media accounts: https://nephrotube.blogspot.com/p/join-nephrotube-on-social-media.html
Travel vaccination in Manchester offers comprehensive immunization services for individuals planning international trips. Expert healthcare providers administer vaccines tailored to your destination, ensuring you stay protected against various diseases. Conveniently located clinics and flexible appointment options make it easy to get the necessary shots before your journey. Stay healthy and travel with confidence by getting vaccinated in Manchester. Visit us: www.nxhealthcare.co.uk
1. ISSN:2161-1068
Mycobacterial Diseases
The International Open Access
Mycobacterial Diseases
Executive Editors
Ying Zhang
Johns Hopkins University, USA
Deepak Kaushal
Tulane University School of Medicine, USA
Subramanian Dhandayuthapani
University of Texas Health Science Center, USA
Gobardhan Das
University of Medicine and Dentistry, USA
David A. Hokey
Aeras, USA
This article was originally published in a journal by OMICS
Publishing Group, and the attached copy is provided by OMICS
Publishing Group for the author’s benefit and for the benefit of
the author’s institution, for commercial/research/educational use
including without limitation use in instruction at your institution,
sending it to specific colleagues that you know, and providing a copy
to your institution’s administrator.
All other uses, reproduction and distribution, including without
limitation commercial reprints, selling or licensing copies or access,
or posting on open internet sites, your personal or institution’s
website or repository, are requested to cite properly.
Available online at: OMICS Publishing Group (www.omicsonline.org)
Digital Object Identifier: http://dx.doi.org/10.4172/2161-1068.1000149
3. Citation: Narh CA, Mosi L, Quaye C, Tay SCK, Bonfoh B, et al. (2014) Genotyping Tools for Mycobacterium ulcerans-Drawbacks and Future Prospects.
J Mycobac Dis 4: 149. doi:10.4172/2161-1068.1000149
Page 2 of 8
Volume 4 • Issue 2 • 1000149
J Mycobac Dis
ISSN: 2161-1068 MDTL, an open access journal
are of immense importance because of their pathogenicity in causing
debilitating ulcers in both animals and humans [17,36-38]. MPMs
are thought to have evolved from M. marinum by the acquisition of a
plasmid, pMUM, which encodes different congeners of mycolactone
(Figure 2), and acquisition of insertion sequences, including IS2404
and IS2606, on both chromosome and plasmid [17,39]. Thus, IS2404
and IS2606 are genetic markers used to differentiate MPMs from
M. marinum M [39,40]. Genetic comparisons of MPM plasmids,
pMUM001 (M. ulcerans), pMUM002 (M. liflandii) and pMUM003
(M. marinum DL), revealed >98% sequence similarity [41]. Primers
targeting genes coding the enoyl reductase (ER) and keto reductase
(KR), enzymes involved in mycolactone synthesis (Figure 2), have been
used in the detection of M. ulcerans, M. liflandii and M. marinum DL
in environmental samples [9]. However, successful differentiation of
MPMs relies on a combination of other genotyping tools (Table 1) and
the use of polymorphic markers including tandem repeat loci (Table 2)
for differentiating the three mentioned species [9,42].
Genome summary and genetic makers for studying
transmission of M. ulcerans
MU is thought to have recently evolved from Mycobacterium
marinum, a fish pathogen, via horizontal transfer of 174 kbp plasmid
and is patho-adapting to the mammalian host [17]. Sequence data
have shown that the two species share >98% sequence homology,
with MU undergoing significant genome reduction [17]. There is
paucity of evolutionary data on MU but phylogenetic analyses of the
different geographical strains have suggested two lineages. The recently
evolved classical lineage, comprises strains from Africa, Australia and
Southeast Asia, and the ancestral lineage include strains from Asia,
provided insights into the necrosis of the soft tissue. These studies
suggested that although bacilli load was relatively high within the
central portions of wounds, there were significant numbers at the
peripherals [23]. Thus, to prevent recurrence of infection, it is
suggested that surgical procedures excise surrounding peripheral
tissues in addition to the necrotic tissues [27]. It takes approximately
6-8 weeks to see visible colonies of MU in pure cultures [29]. Thus,
culture cannot be solely relied on to confirm cases before commencing
treatment [30]. Addition of biochemical and antibiotic susceptibility
tests, on cultured isolates, would be seminal in evaluating efficacy of
existing antimicrobials and reveal emerging drug resistance [30,31]. It
is therefore important for both clinical and research laboratories to set
up antimicrobial surveillance systems in addition to routine diagnosis.
Owing to the delay in culture results and insensitivity of
microscopy, most diagnostic laboratories now rely on PCR or real-time
PCR for prompt and accurate diagnosis [13]. However, the technique;
equipment and reagents, are expensive and cannot be afforded by
poorly resourced laboratories at point of care facilities [32-34]. An
alternative to the latter is Loop-Mediated Isothermal Amplification
(LAMP) which is a novel technique with reported superiority to PCR
in diagnosing M. ulcerans infections [32,33,35]. It is a cost-effective and
robust technique which has the potential to supplant PCR diagnosis
of M. ulcerans infections in poorly resourced laboratories. However,
further studies are needed to optimize it for routine clinical diagnosis.
Signature sequences of Mycolactone Producing Mycobacteria
Mycolactone producing mycobacteria (MPM); Mycobacterium
pseudoshottsii, M. liflandii, M. xenopi, M. marinum DL and M. ulcerans
100 OAgy99
ONM33/04
ONM31/04
ONM22b/0499
Olvory Coast 1
91
100
Clade3
100
99
73
89
100
Togo
Haplotype6
Haplotype5
Haplotype4
Haplotype3
Haplotype2
Clade1
Clade2
5
Benin
Congo
Ivory Coast 2
Angola
66
A
C
B
AwIc
Ga
T
Figure 1: Geographical distribution of African M. ulcerans clades. Map of West-Africa, showing the distribution and SNP haplotypes of three African M.
ulcerans clades. Clade 1: yellow; clade 2: green; clade 3: blue. AW: Amansie West; Ga: strains from the Densu river basin; IC: Ivory Coast; T: Togo; B: Benin;
C: Democratic Republic of Congo; A: Angola. A neighbor-joining tree shows sub grouping of detected haplotypes from the Densu river basin together with the
only strain from Togo into clade 1, strains from AW together with strain Agy99 and strain 1 from the Ivory Coast into clade 2 and all other strains from additional
African countries into clade 3 (scale: number of differences at the SNP loci tested). From Roltgen et al. [34].
4. Citation: Narh CA, Mosi L, Quaye C, Tay SCK, Bonfoh B, et al. (2014) Genotyping Tools for Mycobacterium ulcerans-Drawbacks and Future Prospects.
J Mycobac Dis 4: 149. doi:10.4172/2161-1068.1000149
Page 3 of 8
Volume 4 • Issue 2 • 1000149
J Mycobac Dis
ISSN: 2161-1068 MDTL, an open access journal
of 62.5% [18]. The chromosome has 209 and 83 copies of IS2404
and IS2606 respectively and 4,281 CDS. There are numerous DNA
sequences, within functional and non-functional genes, which have
been employed to differentiate strains of M. ulcerans from other MPMs
[14]. These include variable number tandem repeats (VNTRs) like
locus 6, locus 19, locus 1, ST1, MIRU1 [34,48] and a few housekeeping
genes for Multilocus Sequence Typing (MLST). Despite currently
available tools for studying transmission, it is worth highlighting that
these are still limited compared to malaria and tuberculosis studies for
detecting emergence of resistant strains and carrying out molecular
epidemiological studies [16].
Mexico and South America [43,44]. In one study, Ghanaian MU strains
were suggested to have evolved from a Japanese strain, about 394 to 521
thousand years ago, with subtypes diverging recently [45]. Nonetheless,
there is much genome similarity among all MU strains [17,39,46].
MU, Agy99, has 2 circular replicons, a 5,631,606-bp chromosome
and a 174,155-bp plasmid, pMUM001 [41]. The pMUM001 plasmid
(Figure 2) contains 4 copies of IS2404 and 8 copies of IS2606, both used
as genetic markers [9]. Also prominent are 81 coding domain sequences
(CDS), of which, enoyl reductase (ER) and keto reductase (KR) genes,
two key enzymes involved in biosynthesis of mycolacetone, are used
as genetic markers as well [8,9,47]. It has an average G+C content
Tool Markers Strains Polymorphism Reference
Ribotyping 16S rRNA 3 strains (Australia, Africa and Mexico) Low polymorphism for intra-species differentiation. 16S
rRNA is conserved.
[46]
PFGE and AFLP Whole genome 2 strains (Australia and Africa) Limited by genetic homogeneity of MU strains, and
restriction enzymes
[57]
RFLP IS2404, IS2606 and
polymorphic markers
6 strains (Africa, Australia, Mexico,
Papua New Guinea, Japan and
Suriname)
Low polymorphism for differentiating MU and MPM
isolates from the same origin. Polymorphism depends on
restriction enzymes used.
[40,46]
MLST 8 housekeeping genes 6 strains (Surinam, Papua New Guinea,
Mexico, Japan/China, Africa and Australia)
Polymorphism may vary depending on gene loci [55]
VNTR Over 20 VNTR loci 8 genotypes (Ghana isolates) Variable
for other geographical isolates
Differentiates strains (MU and MPMs) from the same
origin. Genotypes depends on loci used.
[9,14,15,48,85]
SNP ISE, 94 CDS 11 ISE-SNP types 13 SNP haplotypes Strain specific differentiation [45,67]
Table 1: Summary of tools for genotyping M. ulcerans.
0 5000 10000 15000 20000aa Me Me
Me
OH OH
Core
Me
Me
OO
OH
Me
MeMeMe
OH
O
OH
Me
OHOH
MIsA2
module 9module 8
module 7
module 6
module 5
module 4
module 3
module 2
module 1
load
MIsA1
module 7
module 6
module 5
module 4
module 3
module 2
module 1
end
end
load
module 7
module 6
module 5
module 4
module 3
module 2
module 1
load end
module 7
module 6
module 5
module 4
module 3
module 2
module 1
load
module 6
module 5
module 4
module 3
module 2
module 1
load
end
module 6
module 5
module 4
module 3
module 2
module 1
load
end
end
Key: Domain Identification
Ketosynthase load
Ketosynthase
Acyltransferase 1
(malonate)
Acyltransferase 2
(malonate)
Acyltransferase 3
(methylmalonate)
Dehydratase
Enoylreductase
Ketoreductase A
Ketoreductase B
Acyl carrier protein
Inter-modular linker
Integral thioesterase
All pMUM*
M.ulcerans agy99
(origin: human, Africa)
M.ulcerans 753
(origin: human, Japan)
M.ulcerans 98912
(origin: human, China)
M.liflandii 128FXT
(origin: frogs, USA)
M.marinum DL240490
(origin: fish, Israel)
Me
OH
MeMe OH
Me
OHOHO
Me
OH
MeMe OH
Me
OHOHO
Me
OH
MeMe
Me
OHOHO
Me MeMe
Me
OHOH
E
F
OH
O
D
Me
A/B
A/B
MIsB - pMUM002
MIsB - pMUM003
MIsB - MU China
MIsB - MU Japan
MIsB - pMUM001
Figure 2: Genetic organization of the mycolactone biosynthetic cluster from pMUM001, pMUM002 and proposed organization for pMUM003. Mycolactone PKS module
and domain structure is outlined, with the figure key showing the type of domains present in each of the modules. The mycolactone modules are colour coded based
on the type of module responsible for the addition of each two-carbon unit. Shaded modules indicate that the DNA sequence of these regions is unknown or not yet
confirmed. *Organisation of mlsA1 and mlsA2 for all pMUM examined to date, based upon toxin structures. From Pidot et al. [41].
5. Citation: Narh CA, Mosi L, Quaye C, Tay SCK, Bonfoh B, et al. (2014) Genotyping Tools for Mycobacterium ulcerans-Drawbacks and Future Prospects.
J Mycobac Dis 4: 149. doi:10.4172/2161-1068.1000149
Page 4 of 8
Volume 4 • Issue 2 • 1000149
J Mycobac Dis
ISSN: 2161-1068 MDTL, an open access journal
Ribotyping of 16S rRNA gene
The mycobacterial 16S rDNA, which encodes 16S rRNA, is of
great evolutionary importance and has been widely used to trace
phylogenies, differentiate strains of mycobacteria [49-51] and to
resolve ambiguities in bacterial nomenclature [49]. The 16S rRNA
gene is about 1.5 kbp and has both conserved and variable regions
across different taxa [50]. Sequence comparison of this gene between
M. ulcerans and M. marinum showed no significant difference with
slight variation at the 3’-end, between MU strains [51]. Thus, the 16S
rDNA has limited potential for use as a diagnostic marker for MU
infections. Restriction Fragment Length Polymorphism (RFLP) of
the amplified 3’-end however differentiated 29 MU isolates into three
allelic profiles according to three geographical regions; Australia,
Africa and Mexico [46]. It however, could not differentiate between
MU and M. marinum, suggesting its poor adaptability as a tool for
typing genetically related isolates within the same geographical area,
particularly, in epidemiological studies where the specific organism
is to be identified. An alternative to ribotyping, is the conserved rpoB
gene, which encodes the beta subunit of RNA polymerase. It has been
shown to have a higher discriminatory power than 16S rRNA typing [52].
Pulse field gel electrophoresis (PFGE) and Amplified
fragment length polymorphism (AFLP)
PFGE can be used to separate large fragments of DNA in an electric
field, where the voltage switches in three directions. One centrally
and two at 60°, on either sides of the gel. Following restriction digest,
e.g. of a genomic DNA or plasmid, fragments can be run on a gel by
PFGE and banding pattern compared [53]. This technique which
may be used for genotyping or genetic fingerprinting has been used
to compare M. tuberculosis isolates for epidemiological purposes [54].
Prior to the genome sequence of M. ulcerans, PFGE analysis of M.
marinum and M. ulcerans showed a genome size difference of 200 kbp
[55]. In AFLP, fragmented DNA is ligated to specific DNA sequences
(adaptor). Primers complementary to the adaptor are then used to
amplify these fragments. Gel electrophoresis of the amplicons then
shows banding patterns useful for genetic variation studies [56]. AFLP
study of three mycobacteria species classified 12 M. ulcerans isolates
into two geographical types originating from Africa and Australia [57].
Following these findings, coupled with the high sequence similarity
among MU isolates, there has not been much application of these tools
in M. ulcerans genetic studies. Although these tools seem less attractive
to BU researchers, they have been used in other fields for molecular
epidemiological studies [54,56,58,59]. Similar approaches could be
recommended to M. ulcerans research.
IS2404 and IS2606 PCR and RFLP analyses
Sequence analyses of the M. ulcerans reference genome, Agy 99,
have revealed that both IS2404 and IS2606 are present in multiple
copies, on both chromosome and plasmid [41]. Subsequently, primers
targeting these sequences have been designed for use in PCR detection
of M. ulcerans in clinical, veterinary and environmental isolates
[22,25,60,61]. Based on IS2404 RFLP, Chemlal et al. [46] showed that six
distinct genotypes of M. ulcerans could be differentiated corresponding
to the different geographical origins; Africa, Australia, Mexico, Papua
New Guinea, Japan and Suriname. In another study, a typing tool,
IS2426-PCR, was develop to amplify the region between IS2404 and
another MU insertion sequence, IS2606 [40]. Nine different IS2426
PCR genotypes were observed. Furthermore, analysis of the banding
pattern suggested genetic relatedness between MU isolates from
Africa and Southeast Asia [40]. These findings intimate that IS2404
and IS2606 could be useful markers for differentiating geographical
isolates and tracing evolutionary history of MU isolates. However,
their discriminatory abilities for resolving subtle variation between
isolates, within the same geographical region, need to be assessed.
Furthermore, other MPMs have been shown to harbor copies of these
sequences [39,62], suggesting that additional polymorphic markers
are needed to differentiate these species. Therefore, during ecological
studies to source track MU infection within endemic communities, it
is imperative to combine different typing tools in differentiating MU
from other MPMs. Thus, detection of IS2404 and/or IS2606 within
environmental samples should be considered as presumptive detection
of MU. Currently, IS2404 positivity is sufficient to be considered as
definite diagnosis for MU in BU cases, since there have been no reports
of other MPMs causing infection in humans [4] except one recent study
that suggested a propensity for M. pseudoshottsii to cause infection in
humans [42]. Further studies are needed to substantiate the latter.
Multilocus sequence typing (MLST)
In Multilocus sequence typing (MLST), sequences from different
housekeeping genes are compared simultaneously [55]. MLST analysis,
by Stinear et al. [55] using 8 housekeeping genes, on a panel of 18 M.
ulcerans isolates from different geographical areas yielded six different
genotypes, related to the six geographical areas of Surinam, Papua New
Guinea, Mexico, Japan/China, Africa and Australia. This was consistent
with findings by Chemlal et al. [46] using RFLP-IS2404. Further,
comparative genomic studies of M. ulcerans and M. marinum genomes
showed recent divergence of the former from M. marinum, by the
acquisition of a plasmid [55]. While MLST could discriminate between
different geographical isolates and possibly intra-species variation
within the same geographical area, its practicability to routine PCR
reactions in clinical diagnosis would be costly and time consuming.
Published genotypes VNTR Profiles Reference
MIRU1 Locus 6 ST1 Locus 19
M. ulcerans [9,42]
A 1 1 1 2
B 3 1 1 2
C 3 1 2 2
D 1 1 2 2
MMDL
E 1 2 1 2
MLF 1 2 2 1
MPS 1 4 2 2
Amansie West, Ghana [15]
MU strain 1 1 ND 2 ND
MU strain 2 3 ND 1 ND
MU strain 3 3 ND 2 ND
Gh sequence MU strain ND 1 ND 2 [14]
ITM 94-1324 Australian
strain
ND 1 ND 2
ITM 842 Surinam strain ND 1 ND 3
ITM 8756 Japanese
strain
ND 2 ND 4
A, B, C, and D are M. ulcerans designated genotypes, and E is M. marinum DL.
MMDL is M. marinum DL, MPS is M. pseudoshottsii and MLF is M. liflandii. ND, not
done, Gh, Ghana. Work by Ablordey et al. [14] used 9 VNTR markers, excluding
ST1 and MIRU1, but for the purpose of this review, only results for Loci 6 and 19
are shown.
Table 2: VNTR profiles of MU and MPM strain genotypes from published data.
6. Citation: Narh CA, Mosi L, Quaye C, Tay SCK, Bonfoh B, et al. (2014) Genotyping Tools for Mycobacterium ulcerans-Drawbacks and Future Prospects.
J Mycobac Dis 4: 149. doi:10.4172/2161-1068.1000149
Page 5 of 8
Volume 4 • Issue 2 • 1000149
J Mycobac Dis
ISSN: 2161-1068 MDTL, an open access journal
However, MU strain specific primers could be designed and optimized
for use in a multiplex PCR reaction. Development of this tool would
tremendously improve molecular epidemiological studies of MU.
Variable number tandem repeats (VNTR-typing)
Variable number tandem repeats (VNTR) are locations in the
genome where short sequences of DNA occur in a repetitive pattern,
and adjacent to each other, “head-to-tail” [48,63]. These repeats which
vary in number per genome can be used to differentiate between
related species. In M. ulcerans and other MPMs, numerous VNTRs
both within functional and non-functional genes have been identified
at specific loci in the reference genome, Agy 99 [14,48]. PCR reactions
targeting loci like, locus 6, locus 19, MIRU1 and ST1, all with variable
repeats, have successfully been used to differentiate M. ulcerans from
other MPMs [64,65]. It has also been used to resolve the apparent
genetic homogeneity within and between geographical isolates [14].
A study in Ghana, by Hilty et al. [15] using a combination of two
polymorphic VNTR loci, ST1 and MIRU1, on 72 African isolates,
including 57 MU isolates from Ghana, revealed three different
genotypes, with clonal clustering, suggesting genetic diversity of M.
ulcerans in Ghana. In this study, the authors reported 3 strains of MU
in the study area. One strain had a repeat of 2 for ST1 and 1 for MIRU1,
genotype (2,1), and the others had genotypes (1,3) and (2,3) for ST1
and MIRU 1 (Table 2). All repeats were confirmed with sequencing,
suggesting that length polymorphism (50 bp difference of PCR product,
for each locus) has comparable discrimination power as sequence
polymorphism, to reveal intra-species variation in MU. The addition
of other polymorphic loci could therefore increase discrimination
power, revealing more genotypes. This hypothesis was confirmed in a
study, where the authors combined four VNTR markers (Table 2) to
type environmental MU isolates [9]. MU isolates had VNTR profiles
distinct from other MPMs (Table 2). Similarly, a recent study, but
with cultured isolates, corroborated these findings [42]. In elucidating
transmission routes, VNTR typing could be further improved, i.e.
addition of other tandem repeat loci, to source-track MU infections
to specific risk environments. Thus, it is possible to compare VNTR
profiles of human isolates to environmental isolates, within endemic
communities, to trace reservoirs and vectors of MU [9,42]. To increase
efficiency of VNTR as a genotyping tool, a Multilocus VNTR analysis
(MLVA) can be adapted and automated for typing MU strains. With
this technique, PCR primers (in a multiplex PCR ) for the different loci
are labelled with different fluorescent dyes and products separated by
capillary electrophoresis using an automated sequencer [63]. MLVA
has become the reference typing method for M. tuberculosis [66].
Single nucleotide polymorphisms (SNPs) and microarray
analysis
Single Nucleotide Polymorphism (SNP typing), detects a single
base pair mutation at a specific locus, revealing genetic variations
between members of a species. Using this tool, various techniques from
hybridization to enzyme-based methods and sequencing have been
employed to reveal subtle differences in seemingly identical strains.
Kaser et al. [67] employed SNP analysis of the IS2404, to genotype 83
M. ulcerans isolates from African countries. They identified 11 ISE-
SNP types that differentiated regional strains into three haplotypes
[67]. A similar study by the same group, using 94 protein coding genes,
differentiated 54 Ghanaian MU isolates into 13 SNP haplotypes [45].
Following these findings, Roltgen et al. [34] developed a real-time PCR
SNP typing method to genotype M. ulcerans patient isolates, collected
from different parts of Ghana, and other isolates from Cote d’ Ivoire,
Democratic Republic of Congo, Benin, Togo and Angola. Using 65 SNP
loci, the authors identified nine haplotypes for the Ghanaian isolates.
Neighbor-joining tree grouped isolates into three clades (Figure 1). Six
haplotypes around the Densu River, southern Ghana, and isolates from
Togo formed a clade. Isolates from central Ghana (three haplotypes)
clustered with one Ivoirian isolate. The other African isolates formed
the third clade. In one case, the authors observed dominance of one
clonal complex and its clustering along the Densu River, suggesting
possible focal transmission. Large scale application of SNP typing for
epidemiological studies would involve the use of DNA microarrays on
chips [68]. This technique has been used to differentiate M. tuberculosis
from M. bovis BCG, through the identification of 18 regions of
diversity (RD1-RD18) [69,70]. Although, this technique is costly, it is
perhaps the most informative genotyping tool for detecting mutants
in the population and picking up drug resistant strains. Reference
laboratories could develop capacity in applying this tool to studying the
population structure of MU, its evolutionary history and in monitoring
the emergence of drug resistant strains.
Modification of fine typing tools for M. ulcerans research
Advances in molecular biology, genomics, proteomics and
bioinformatics have greatly accelerated research in understanding
the molecular mechanisms of infection, pathology and treatment of
various mycobacterial diseases, notably, TB and BU [2]. Sequencing
of the genome of MU strain Agy99 has significantly improved studies
on diagnosis and transmission [55]. However, these tools are currently
inadequate in addressing critical research questions including the
mode of transmission and hence the development of strategies for
preventing and controlling the disease. A greater burden lies on
molecular biologists and geneticists to develop finer typing tools for
diagnosis and transmission studies. There is the need to rescan the
published genomes of reference strains for nuanced sequences that
could be used to differentiate between MU strains and other MPMs.
Supplementary studies could explore the bacteria population structure
within the different geographical areas, strain variation, and perceive
how M. ulcerans may be evolving from an environmental niche
to the human host. To achieve these, molecular epidemiological
studies should not focus only on genotyping human MU isolates
but need to also genotype environmental MU isolates. As previously
discussed VNTR typing shows variation in environmental isolates
[9] supporting SNP data that suggest clustering of a clonal complex
within certain river basins [34]. Evidently, typing of more isolates,
both human and environmental, would provide better insight into the
bacteria population structure, which strains are niche adapting to the
mammalian host, and the relative virulence of these strains, as inferred
from mycolactone studies [71]. Recent VNTR typing of cultured MU
isolates suggest that certain strains to be more readily cultured and/or
transmissible than others [42]. Application of MLVA may increase the
discriminatory power of VNTR typing. More specific primers targeting
potential polymorphic markers, e.g. regions of difference (RD) [72],
need to be designed to complement existing repertoire in diagnosis
and for molecular epidemiological studies. Bioinformatics capacity and
sequence data accessibility are strongly recommended to facilitate this.
Additionally, high-throughput sequencing including next-generation
sequencing and pyrosequencing, may improve quality of sequence data
for SNP analysis [45]. RFLP-PCR would enhance strain differentiation
if combined with capillary electrophoresis [63]. One important
challenge however will be the standardization of genotyping protocols
as most typing tools for MU give different genetic profiles. This could
lead to varying designation of genotypes by different researchers and
7. Citation: Narh CA, Mosi L, Quaye C, Tay SCK, Bonfoh B, et al. (2014) Genotyping Tools for Mycobacterium ulcerans-Drawbacks and Future Prospects.
J Mycobac Dis 4: 149. doi:10.4172/2161-1068.1000149
Page 6 of 8
Volume 4 • Issue 2 • 1000149
J Mycobac Dis
ISSN: 2161-1068 MDTL, an open access journal
laboratories. We suggest that researchers develop and standardize
genotyping protocols to facilitate easy identification and comparison
of strains.
Surveillance for emerging drug resistance
Rifampicin and streptomycin are the two WHO recommended
antimicrobials that in combination are used for treating MU infections.
Rifampicin inhibits bacteria DNA synthesis by inhibiting bacterial
DNA-dependent RNA polymerase [73]. Streptomycin inhibits protein
synthesis by binding to the bacterial 16S rRNA, interfering with the
binding of formyl-methionyl-tRNA to the 30S subunit [74]. There have
been reported cases of drug resistance to rifampicin in a mouse model
[75] and in clinical isolates [76,77]. In the murine study, three MU
mutants were isolated after rifampicin monotherapy. Molecular data
showed that mutants harbored Ser416Phe and His420Tyr mutations in
the rpoB gene [75]. Previous studies have shown that these mutations
in the rpoB gene, confers rifampicin resistance to tubercle bacilli [73].
In another study, using sequence-based approach, a point mutation
was detected in the rpoB gene at codon Ser522 leading to an amino
acid change in 0.9% of the clinical isolates used [76]. Similarly,
resistance to streptomycin has been reported in the closely related
species, M. tuberculosis, in the rpsL gene [78]. These resistances could
be compounded in cases where additional antibiotics are prescribed for
the treatment of secondary infections, in ulcerative lesions [79].
These suggest that some MU strains have the propensity to develop
resistance against these effective drugs. Current available tools for
genotyping MU need to be improved and adapted for detecting
resistant strains. SNP typing holds tremendous potential for this
purpose. SNP typing been applied in various fields to detect drug
resistant strains [76,78,80-83]. This technique in combination with
VNTR typing could be used to determine strains that are becoming
resistant. However, these techniques need to be tailored, obviating
the need for expensive equipment, for use in developing countries,
and at point of care facilities, where majority of cases are detected and
treated. Additionally, MU isolates could be screened for susceptibility
to existing antimicrobials, as complementary to the currently used ones
or as alternatives, in the event of future resistance. New drugs with
different modes of action could also be developed to combat future
resistant strains [84].
Conclusion
M. ulcerans infection is the third most common mycobacterial
infections after tuberculosis and leprosy, however, it has received
comparably less attention. Available tools for genotyping strains of
the pathogen have shown low discriminatory power in resolving
intra-species variation, particularly at regional levels. The application
of finer genotyping tools including VNTR and SNP analysis, have
shown heterogeneity within isolates from the same geographical area.
Although these strains share nearly identical sequences, rescanning
of the genome for nuanced sequences, refinement of existing tools
and modification of tools from other fields, particularly TB, may help
uncover other pathogenic subtypes. Concurrently, such advancements
may further help detect emerging drug resistant strains. In the wake
of increasing cases of Buruli ulcer, cumulative efforts including
improvement in diagnostic methods and fine-tuning of genotyping
tools are key to elucidating transmission routes, studying the molecular
epidemiology of MU and detection of incipient resistant strains.
Acknowledgement
This paper is partially based on work supported by the consortium Afrique One
“Ecosystem and Population Health: “Expanding Frontiers in Health”. Afrique One is
funded by the Wellcome Trust (WT087535MA). Charles Narh was also supported
by CODESRIA’s Small Grant for Thesis Writing (Ref SGRT.47/T12).
References
1. George KM, Pascopella L, Welty DM, Small PL (2000) A Mycobacterium
ulcerans toxin, mycolactone, causes apoptosis in guinea pig ulcers and tissue
culture cells. Infect Immun 68: 877-883.
2. Walsh DS, Portaels F, Meyers WM (2010) Recent advances in leprosy and
Buruli ulcer (Mycobacterium ulcerans infection). CurrOpin Infect Dis 23: 445-
455.
3. WHO (2010). Number of new cases of Buruli ulcer reported, 2010. apps.who.
int: WHO.
4. (2008) Buruli ulcer: progress report, 2004-2008. WklyEpidemiol Rec 83: 145-
154.
5. Debacker M, Portaels F, Aguiar J, Steunou C, Zinsou C, et al. (2006) Risk
factors for Buruli ulcer, Benin. Emerg Infect Dis 12: 1325-1331.
6. Pouillot R, Matias G, Wondje CM, Portaels F, Valin N, et al. (2007) Risk factors
for buruli ulcer: a case control study in Cameroon. PLoS Negl Trop Dis 1: e101.
7. Raghunathan PL, Whitney EA, Asamoa K, Stienstra Y, Taylor TH Jr, et al.
(2005) Risk factors for Buruli ulcer disease (Mycobacterium ulcerans Infection):
results from a case-control study in Ghana. Clin Infect Dis 40: 1445-1453.
8. Williamson HR, Benbow ME, Campbell LP, Johnson CR, Sopoh G, et al. (2012)
Detection of Mycobacterium ulcerans in the environment predicts prevalence of
Buruli ulcer in Benin. PLoS Negl Trop Dis 6: e1506.
9. Williamson HR, Benbow ME, Nguyen KD, Beachboard DC, Kimbirauskas RK,
et al. (2008) Distribution of Mycobacterium ulcerans in buruli ulcer endemic and
non-endemic aquatic sites in Ghana. PLoS Negl Trop Dis 2: e205.
10. Johnson PD, Lavender CJ (2009) Correlation between Buruli ulcer and vector-
borne notifiable diseases, Victoria, Australia. Emerg Infect Dis 15: 614-615.
11. Portaels F, Elsen P, Guimaraes-Peres A, Fonteyne PA, Meyers WM (1999)
Insects in the transmission of Mycobacterium ulcerans infection. Lancet 353:
986.
12. Benbow ME, Williamson H, Kimbirauskas R, McIntosh MD, Kolar R, et al.
(2008) Aquatic invertebrates as unlikely vectors of Buruli ulcer disease. Emerg
Infect Dis 14: 1247-1254.
13. Yeboah-Manu D, Danso E, Ampah K, Asante-Poku A, Nakobu Z, et al. (2011)
Isolation of Mycobacterium ulcerans from swab and fine-needle-aspiration
specimens. J Clin Microbiol 49: 1997-1999.
14. Ablordey A, Hilty M, Stragier P, Swings J, Portaels F (2005) Comparative
nucleotide sequence analysis of polymorphic variable-number tandem-repeat
Loci in Mycobacterium ulcerans. J Clin Microbiol 43: 5281-5284.
15. Hilty MD, Yeboah-Manu, Boakye D, Mensah-Quainoo E, Rondini S, et al.
(2006) Genetic diversity in Mycobacterium ulcerans isolates from Ghana
revealed by a newly identified locus containing a variable number of tandem
repeats. J Bacteriol 188: 1462-1465.
16. Merritt RW, Walker ED, Small PL, Wallace JR, Johnson PD, et al. (2010)
Ecology and transmission of Buruli ulcer disease: a systematic review. PLoS
Negl Trop Dis 4: e911.
17. Stinear TP, Seemann T, Pidot S, Frigui W, Reysset G, et al. (2007) Reductive
evolution and niche adaptation inferred from the genome of Mycobacterium
ulcerans, the causative agent of Buruli ulcer. Genome Res 17: 192-200.
18. Stinear TP, Seemann T, Pidot S, Frigui W, Reysset G, et al. (2007) Reductive
evolution and niche adaptation inferred from the genome of Mycobacterium
ulcerans, the causative agent of Buruli ulcer. Genome Res 17: 192-200.
19. Radomski N, Cambau E, Moulin L, Haenn S, Moilleron R, et al. (2010)
Comparison of culture methods for isolation of nontuberculous mycobacteria
from surface waters. Appl Environ Microbiol 76: 3514-3520.
20. Ross BC, Johnson PD, Oppedisano F, Marino L, Sievers A, et al. (1997)
Detection of Mycobacterium ulcerans in environmental samples during an
outbreak of ulcerative disease. Appl Environ Microbiol 63: 4135-4138.
8. Citation: Narh CA, Mosi L, Quaye C, Tay SCK, Bonfoh B, et al. (2014) Genotyping Tools for Mycobacterium ulcerans-Drawbacks and Future Prospects.
J Mycobac Dis 4: 149. doi:10.4172/2161-1068.1000149
Page 7 of 8
Volume 4 • Issue 2 • 1000149
J Mycobac Dis
ISSN: 2161-1068 MDTL, an open access journal
21. Addo P, Adu-Addai B, Quartey M, Abbas M, Okang I, et al. (2007) Clinical
and histopathological presentation of Buruli ulcer in experimentally infected
grasscutters (Thryonomysswinderianus). Internet J. Trop. Med. 3:e2.
22. Phillips R, Horsfield C, Kuijper S, Lartey A, Tetteh I, et al. (2005) Sensitivity
of PCR targeting the IS2404 insertion sequence of Mycobacterium ulcerans
in an Assay using punch biopsy specimens for diagnosis of Buruli ulcer. J
ClinMicrobiol 43: 3650-3656.
23. Rondini S, Horsfield C, Mensah-Quainoo E, Junghanss T, Lucas S, et al.
(2006) Contiguous spread of Mycobacterium ulcerans in Buruli ulcer lesions
analysed by histopathology and real-time PCR quantification of mycobacterial
DNA. J Pathol 208: 119-128.
24. Walsh DS, Meyers WM, Portaels F, Lane JE, Mongkolsirichaikul D, et al. (2005)
High rates of apoptosis in human Mycobacterium ulcerans culture-positive
buruli ulcer skin lesions. Am J Trop Med Hyg 73: 410-415.
25. Yeboah-Manu D, Asante-Poku A, Asan-Ampah K, Ampadu ED, Pluschke G
(2011) Combining PCR with microscopy to reduce costs of laboratory diagnosis
of Buruli ulcer. Am J Trop Med Hyg 85: 900-904.
26. Beissner M, Herbinger KH, Bretzel G (2010) Laboratory diagnosis of Buruli
ulcer disease. Future Microbiol 5: 363-370.
27. Bretzel G, Siegmund V, Racz P, van Vloten F, Ngos F, et al. (2005) Post-
surgical assessment of excised tissue from patients with Buruli ulcer disease:
progression of infection in macroscopically healthy tissue. Trop Med Int Health
10: 1199-1206.
28. Herbinger KH, Adjei O, Awua-Boateng NY, Nienhuis WA, Kunaa L, et al.
(2009) Comparative study of the sensitivity of different diagnostic methods for
the laboratory diagnosis of Buruli ulcer disease. Clin Infect Dis 48: 1055-1064.
29. Grange JM (1996) The biology of the genus Mycobacterium. Soc Appl Bacteriol
Symp Ser 25: 1S-9S.
30. WHO/CDS/CPE/GBUI. (2001). Buruli ulcer, Mycobacterium ulcerans infection.
In K. Asiedu, R. Scherpbier, and M. Raviglione (Eds.): WHO.
31. Kondo M, Kurokawa I, Ito Y, Yamanaka K, Yamazaki T, et al. (2009) Leg ulcer
caused by Mycobacterium ulcerans ssp. shinshuense infection. Int J Dermatol
48: 1330-1333.
32. Ablordey A, Amissah DA, Aboagye IF, Hatano B, Yamazaki T, et al. (2012)
Detection of Mycobacterium ulcerans by the loop mediated isothermal
amplification method. PLoS Negl Trop Dis 6: e1590.
33. de Souza DK, Quaye C, Mosi L, Addo P, Boakye DA (2012) A quick and cost
effective method for the diagnosis of Mycobacterium ulcerans infection. BMC
Infect Dis 12: 8.
34. Röltgen K, Qi W, Ruf MT, Mensah-Quainoo E, Pidot SJ, et al. (2010) Single
nucleotide polymorphism typing of Mycobacterium ulcerans reveals focal
transmission of buruli ulcer in a highly endemic region of Ghana. PLoS Negl
Trop Dis 4: e751.
35. Njiru ZK, Yeboah-Manu D, Stinear TP, Fyfe JA (2012) Rapid and sensitive
detection of Mycobacterium ulcerans by use of a loop-mediated isothermal
amplification test. J Clin Microbiol 50: 1737-1741.
36. Mve-Obiang A, Lee RE, Umstot ES, Trott KA, Grammer TC, et al. (2005). A
newly discovered mycobacterial pathogen isolated from laboratory colonies of
Xenopus species with lethal infections produces a novel form of mycolactone,
the Mycobacterium ulcerans macrolide toxin. Infect Immun 73: 3307-3312.
37. Rhodes MW, Kator H, Kotob S, van Berkum P, Kaattari I, et al. (2001) A
unique Mycobacterium species isolated from an epizootic of striped bass
(Moronesaxatilis). Emerg Infect Dis 7: 896-899.
38. Broussard GW, Ennis DG (2007) Mycobacterium marinum produces long-
term chronic infections in medaka: a new animal model for studying human
tuberculosis. CompBiochemPhysiol C Toxicol Pharmacol 145: 45-54.
39. Tobias NJ, Doig KD, Medema MH, Chen H, Haring V, et al. (2013) Complete
genome sequence of the frog pathogen Mycobacterium ulceransecovarLiflandii.
J Bacteriol 195: 556-564.
40. Stinear T, Davies JK, Jenkin GA, Portaels F, Ross BC, et al. (2000) A simple
PCR method for rapid genotype analysis of Mycobacterium ulcerans. J Clin
Microbiol 38: 1482-1487.
41. Pidot SJ, Hong H, Seemann T, Porter JL, Yip MJ, et al. (2008) Deciphering
the genetic basis for polyketide variation among mycobacteria producing
mycolactones. BMC Genomics 9: 462.
42. Williamson H, Phillips R, Sarfo S, Wansbrough-Jones M, Small P (2014)
Genetic Diversity of PCR-Positive, Culture-Negative and Culture-Positive
Mycobacterium ulcerans Isolated from Buruli Ulcer Patients in Ghana. PLoS
ONE 9(2) e88007.
43. Rondini S, Käser M, Stinear T, Tessier M, Mangold C, et al. (2007) Ongoing
genome reduction in Mycobacterium ulcerans. Emerg Infect Dis 13: 1008-1015.
44. Käser M, Rondini S, Naegeli M, Stinear T, Portaels F, et al. (2007) Evolution
of two distinct phylogenetic lineages of the emerging human pathogen
Mycobacterium ulcerans. BMC Evol Biol 7: 177.
45. Qi W, Käser M, Röltgen K, Yeboah-Manu D, Pluschke G (2009) Genomic
diversity and evolution of Mycobacterium ulcerans revealed by next-generation
sequencing. PLoS Pathog 5: e1000580.
46. Chemlal K, Huys G, Fonteyne PA, Vincent V, Lopez AG, et al. (2001) Evaluation
of PCR-restriction profile analysis and IS2404 restriction fragment length
polymorphism and amplified fragment length polymorphism fingerprinting for
identification and typing of Mycobacterium ulcerans and M. marinum. J Clin
Microbiol 39(9) 3272-8.
47. Fyfe JA, Lavender CJ, Johnson PD, Globan M, Sievers A, et al. (2007)
Development and application of two multiplex real-time PCR assays for the
detection of Mycobacterium ulcerans in clinical and environmental samples.
Appl Environ Microbiol 73: 4733-4740.
48. Ablordey A, Swings J, Hubans C, Chemlal K, Locht C, et al. (2005) Multilocus
variable-number tandem repeat typing of Mycobacterium ulcerans. J Clin
Microbiol 43: 1546-1551.
49. Janda JM, Abbott SL (2007) 16S rRNA gene sequencing for bacterial
identification in the diagnostic laboratory: pluses, perils, and pitfalls. J Clin
Microbiol 45: 2761-2764.
50. Clarridge JE 3rd (2004) Impact of 16S rRNA gene sequence analysis for
identification of bacteria on clinical microbiology and infectious diseases. Clin
Microbiol Rev 17: 840-862, table of contents.
51. Portaels F, Fonteyene PA, de Beenhouwer H, de Rijk P, Guédénon A, et al.
(1996) Variability in 3’ end of 16S rRNA sequence of Mycobacterium ulcerans
is related to geographic origin of isolates. J Clin Microbiol 34: 962-965.
52. De Clerck E, De Vos P (2004) Genotypic diversity among Bacillus licheniformis
strains from various sources. FEMS Microbiol Lett 231: 91-98.
53. Schwartz DC, Cantor CR (1984) Separation of yeast chromosome-sized DNAs
by pulsed field gradient gel electrophoresis. Cell 37: 67-75.
54. Singh SP, Salamon H, Lahti CJ, Farid-Moyer M, Small PM (1999) Use of
pulsed-field gel electrophoresis for molecular epidemiologic and population
genetic studies of Mycobacterium tuberculosis. J Clin Microbiol 37: 1927-1931.
55. Stinear TP, Jenkin GA, Johnson PD, Davies JK (2000) Comparative genetic
analysis of Mycobacterium ulcerans and Mycobacterium marinum reveals
evidence of recent divergence. J Bacteriol 182: 6322-6330.
56. Vos P, Hogers R, Bleeker M, Reijans M, van de Lee T, et al. (1995) AFLP: a
new technique for DNA fingerprinting. Nucleic Acids Res 23: 4407-4414.
57. Huys G, Rigouts L, Chemlal K, Portaels F, Swings J (2000) Evaluation of
Amplified Fragment Length Polymorphism Analysis for Inter- and Intraspecific
Differentiation ofMycobacteriumbovis, M. tuberculosis, andM. ulcerans. Journal
of Clinical Microbiology 38(10) 3675-3680.
58. Vijayachari P, Ahmed N, Sugunan AP, Ghousunnissa S, Rao KR, et al. (2004)
Use of fluorescent amplified fragment length polymorphism for molecular
epidemiology of leptospirosis in India. J Clin Microbiol 42: 3575-3580.
59. Zhong D, Temu EA, Guda T, Gouagna L, Menge D, et al. (2006) Dynamics of
gene introgression in the African malaria vector Anopheles gambiae. Genetics
172: 2359-2365.
60. Stinear T, Davies JK, Jenkin GA, Hayman JA, Oppedisano F, et al. (2000)
Identification of Mycobacterium ulcerans in the environment from regions in
Southeast Australia in which it is endemic with sequence capture-PCR. Appl
Environ Microbiol 66: 3206-3213.
61. Ranger BS, Mahrous EA, Mosi L, Adusumilli S, Lee RE, et al. (2006) Globally
distributed mycobacterial fish pathogens produce a novel plasmid-encoded
toxic macrolide, mycolactone F. Infect Immun 74: 6037-6045.
62. Suykerbuyk P, Vleminckx K, Pasmans F, Stragier P, Ablordey A, et al. (2007)
Mycobacterium liflandii infection in European colony of Siluranatropicalis.
Emerg Infect Dis 13: 743-746.
9. Citation: Narh CA, Mosi L, Quaye C, Tay SCK, Bonfoh B, et al. (2014) Genotyping Tools for Mycobacterium ulcerans-Drawbacks and Future Prospects.
J Mycobac Dis 4: 149. doi:10.4172/2161-1068.1000149
Page 8 of 8
Volume 4 • Issue 2 • 1000149
J Mycobac Dis
ISSN: 2161-1068 MDTL, an open access journal
63. Li W, Raoult D, Fournier PE (2009) Bacterial strain typing in the genomic era.
FEMS Microbiol Rev 33: 892-916.
64. Stragier P, Ablordey A, Durnez L, Portaels F (2007) VNTR analysis
differentiates Mycobacterium ulcerans and IS2404 positive mycobacteria. Syst
Appl Microbiol 30: 525-530.
65. Lavender CJ, Stinear TP, Johnson PD, Azuolas J, Benbow ME, et al. (2008)
Evaluation of VNTR typing for the identification of Mycobacterium ulcerans in
environmental samples from Victoria, Australia. FEMS Microbiol Lett 287: 250-
255.
66. Le Flèche P, Fabre M, Denoeud F, Koeck JL, Vergnaud G (2002) High
resolution, on-line identification of strains from the Mycobacterium tuberculosis
complex based on tandem repeat typing. BMC Microbiol 2: 37.
67. Käser M, Hauser J, Pluschke G (2009) Single nucleotide polymorphisms on
the road to strain differentiation in Mycobacterium ulcerans. J Clin Microbiol
47: 3647-3652.
68. Käser M, Pluschke G (2008) Differential gene repertoire in Mycobacterium
ulcerans identifies candidate genes for patho-adaptation. PLoS Negl Trop Dis
2: e353.
69. Behr MA, Wilson MA, Gill WP, Salamon H, Schoolnik GK, et al. (1999)
Comparative genomics of BCG vaccines by whole-genome DNA microarray.
Science 284: 1520-1523.
70. Gordon SV, Brosch R, Billault A, Garnier T, Eiglmeier K, et al. (1999)
Identification of variable regions in the genomes of tubercle bacilli using
bacterial artificial chromosome arrays. Mol Microbiol 32: 643-655.
71. Hong H, Demangel C, Pidot SJ, Leadlay PF, Stinear T (2008) Mycolactones:
immunosuppressive and cytotoxic polyketides produced by aquatic
mycobacteria. Nat Prod Rep 25: 447-454.
72. Huber CA, Ruf MT, Pluschke G, Käser M (2008) Independent loss of
immunogenic proteins in Mycobacterium ulcerans suggests immune evasion.
Clin Vaccine Immunol 15: 598-606.
73. Honore N, Cole ST (1993) Molecular basis of rifampin resistance in
Mycobacterium leprae. Antimicrob Agents Chemother 37: 414-418.
74. Sharma D, Cukras AR, Rogers EJ, Southworth DR, Green R (2007) Mutational
analysis of S12 protein and implications for the accuracy of decoding by the
ribosome. J Mol Biol 374: 1065-1076.
75. Marsollier L, Honore N, Legras P, Manceau AL, Kouakou H, et al. (2003).
Isolation of three Mycobacterium ulcerans strains resistant to rifampin after
experimental chemotherapy of mice.Antimicrob Agents Chemother 47: 1228-
1232.
76. Beissner M, Awua-Boateng NY, Thompson W, Nienhuis WA, Klutse E, et al.
(2010) A genotypic approach for detection, identification, and characterization
of drug resistance in Mycobacterium ulcerans in clinical samples and isolates
from Ghana. Am J Trop Med Hyg 83: 1059-1065.
77. Jansson M, Beissner M, Phillips RO, Badziklou K, Piten E, et al. (2014)
Comparison of two assays for molecular determination of rifampin resistance
in clinical samples from patients with buruli ulcer disease. J ClinMicrobiol 52:
1246-1249.
78. Das R, Gupta P, Singh P, Chauhan DS, Katoch K, et al. (2009) Association of
mutations in rpsL gene with high degree of streptomycin resistance in clinical
isolates of Mycobacterium tuberculosis in India. Indian J Med Res 129: 108-
110.
79. Yeboah-Manu D, Kpeli GS, Ruf MT, Asan-Ampah K, Quenin-Fosu K, et al.
(2013) Secondary bacterial infections of buruli ulcer lesions before and after
chemotherapy with streptomycin and rifampicin. PLoS Negl Trop Dis 7: e2191.
80. Alam MT, de Souza DK, Vinayak S, Griffing SM, Poe AC, et al. (2011) Selective
sweeps and genetic lineages of Plasmodium falciparum drug -resistant alleles
in Ghana. J Infect Dis 203: 220-227.
81. Brown-Elliott BA, Nash KA, Wallace RJ Jr (2012) Antimicrobial susceptibility
testing, drug resistance mechanisms, and therapy of infections with
nontuberculous mycobacteria. Clin Microbiol Rev 25: 545-582.
82. Miller EN, Jamieson SE, Joberty C, Fakiola M, Hudson D, et al. (2004) Genome-
wide scans for leprosy and tuberculosis susceptibility genes in Brazilians.
Genes Immun 5: 63-67.
83. Palomino JC (2009) Molecular detection, identification and drug resistance
detection in Mycobacterium tuberculosis. FEMS Immunol Med Microbiol 56:
103-111.
84. Hernandez V, Crépin T, Palencia A, Cusack S, Akama T, et al. (2013)
Discovery of a novel class of boron-based antibacterials with activity against
gram-negative bacteria. Antimicrob Agents Chemother 57: 1394-1403.
85. Hilty M, Käser M, Zinsstag J, Stinear T, Pluschke G (2007) Analysis of the
Mycobacterium ulcerans genome sequence reveals new loci for variable
number tandem repeats (VNTR) typing. Microbiology 153: 1483-1487.
Citation: Narh CA, Mosi L, Quaye C, Tay SCK, Bonfoh B, et al. (2014)
Genotyping Tools for Mycobacterium Ulcerans Drawbacks and Future
Prospects. J Mycobac Dis 4: 149. doi:10.4172/2161-1068.1000149
Submit your next manuscript and get advantages of OMICS
Group submissions
Unique features:
• User friendly/feasible website-translation of your paper to 50 world’s leading languages
• Audio Version of published paper
• Digital articles to share and explore
Special features:
• 300 Open Access Journals
• 25,000 editorial team
• 21 days rapid review process
• Quality and quick editorial, review and publication processing
• Indexing at PubMed (partial), Scopus, EBSCO, Index Copernicus and Google Scholar etc
• Sharing Option: Social Networking Enabled
• Authors, Reviewers and Editors rewarded with online Scientific Credits
• Better discount for your subsequent articles
Submit your manuscript at: http://www.omicsonline.org/submission