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1. Identification of genes involved in the intracellular survival of Mycobacterium tuberculosis

Author

Hobson, Russell James

Subjects
610, Mycobacteria
Abstract

Little is known about the survival mechanics of Mycobacterium tuberculosis within the human host. A number of genes are likely to be activated during infection, especially within macrophages, and these potential virulence determinants will be of great importance in the ultimate development of new vaccines and antimycobacterials. In an effort to identify some of these virulence determinants, a M.tuberculosis H37Rv gene promoter-probe library in a BCG host was constructed using an E.coli - mycobacterial shuttle vector with lacZ as the reporter gene. 4800 individual clones were arrayed in a 96 well microtitre(TM) format, enabling the testing of individual clones for promoter activity under a variety of environmental stressing conditions. Two separate in vitro screenings of the arrayed clones identified 53 clones that exhibited up-regulation of lacZ when stressed. 50 of these clones were bi-directionally sequenced to determine the cloned M.tuberculosis H37Rv DNA sequence. Identification of the location of the cloned DNA within the M.tuberculosis H37Rv genome identified 32 clones likely to containing potential promoter sequences. By measuring the levels of lacZ expression post-infection of macrophages, compared to in vitro expression, clones were identified which contained potential promoter sequences up-regulated within the macrophage. Quantitative Reverse Transcriptase-PCR was employed, using cDNA obtained from M.tuberculosis from infected and un-infected macrophages, to measure the levels of expression of 5 M.tuberculosis H37Rv genes identified as potentially up- regulated during infection. The results suggest that two of the five genes, genes Rv1265 and Rv2711, are indeed up-regulated during infection. The protein coded for by the gene Rv1265 has an unknown function whilst the gene Rv2711 encodes the protein IdeR, which is an iron-dependent repressor. Sequence and lacZ expression analysis identified four independently selected and screened clones which contained near identical sequences for the same promoter - that of the gene Rv0440 (groEL2). Gene Rv0440 codes for a 60KDa heat shock protein previously identified as up-regulated during infection. The identification of four independent clones containing the promoter for this gene validates this technology as a means of identifying M.tuberculosis H37Rv genes up-regulated during infection.

Published
2000

3. Use of an arrayed promoter-probe library for the identification of macrophage-regulated genes in Mycobacterium tuberculosis

Author

Hobson, Russell J., McBride, Alan J. A., Kempsell, Karen E., and Dale, Jeremy W.

Subjects
Microbiological research -- Analysis, Macrophages -- Genetic aspects, Genetic regulation -- Physiological aspects, Mycobacterium tuberculosis -- Genetic aspects, DNA -- Genetic aspects, Infection -- Microbiology, Biological sciences
Abstract

Research has been conducted on Mycobacterium tuberculosis survival within the human host after infection. The arrayed library of M. tuberculosis DNA fragments has been constructed in the promoter-probe vector and used for the screening of promoters responding to the environmental conditions.

Published
2002

4. Integrated ecological monitoring in Wales: the Glastir Monitoring and Evaluation Programme field survey.

Author

Wood, Claire M., Alison, Jamie, Botham, Marc S., Burden, Annette, Edwards, Francois, Garbutt, R. Angus, George, Paul B. L., Henrys, Peter A., Hobson, Russell, Jarvis, Susan, Keenan, Patrick, Keith, Aidan M., Lebron, Inma, Maskell, Lindsay C., Norton, Lisa R., Robinson, David A., Seaton, Fiona M., Scarlett, Peter, Siriwardena, Gavin M., and Skates, James

Subjects
PHYSICAL geography, ENVIRONMENTAL monitoring, LAND cover, PHYSICAL geology, INSECT pollinators, DIGITAL Object Identifiers, CLIMATE change mitigation
Abstract

The Glastir Monitoring and Evaluation Programme (GMEP) ran from 2013 until 2016, and was probably the most comprehensive programme of ecological study ever undertaken at a national scale in Wales. The programme aimed to (1) set up an evaluation of the environmental effects of the Glastir agri-environment scheme and (2) quantify environmental status and trends across the wider countryside of Wales. The focus was on outcomes for climate change mitigation, biodiversity, soil and water quality, woodland expansion and cultural landscapes. As such, GMEP included a large field survey component, collecting data on a range of elements including vegetation, land cover and use, soils, freshwaters, birds and insect pollinators from up to 300 1 km squares throughout Wales. The field survey capitalised upon the UKCEH Countryside Survey of Great Britain, which has provided an extensive set of repeated, standardised ecological measurements since 1978. The design of both GMEP and the UKCEH Countryside Survey involved stratified-random sampling of squares from a 1 km grid, ensuring proportional representation from land classes with distinct climate, geology and physical geography. Data were collected from different land cover types and landscape features by trained professional surveyors, following standardised and published protocols. Thus, GMEP was designed so that surveys could be repeated at regular intervals to monitor the Welsh environment, including the impacts of agri-environment interventions. One such repeat survey is scheduled for 2021 under the Environment and Rural Affairs Monitoring and Modelling Programme (ERAMMP). Data from GMEP have been used to address many applied policy questions, but there is major potential for further analyses. The precise locations of data collection are not publicly available, largely for reasons of landowner confidentiality. However, the wide variety of available datasets can be (1) analysed at coarse spatial resolutions and (2) linked to each other based on square-level and plot-level identifiers, allowing exploration of relationships, trade-offs and synergies. This paper describes the key sets of raw data arising from the field survey at co-located sites, 2013 to 2016. Data from each of these survey elements are available with the following Digital Object Identifiers. Landscape features, https://doi.org/10.5285/82c63533-529e-47b9-8e78-51b27028cc7f, https://doi.org/10.5285/9f8d9cc6-b552-4c8b-af09-e92743cdd3de, https://doi.org/10.5285/f481c6bf-5774-4df8-8776-c4d7bf059d40; Vegetation plots, https://doi.org/10.5285/71d3619c-4439-4c9e-84dc-3ca873d7f5cc; Topsoil physico-chemical properties, https://doi.org/10.5285/0fa51dc6-1537-4ad6-9d06-e476c137ed09; Topsoil meso-fauna, https://doi.org/10.5285/1c5cf317-2f03-4fef-b060-9eccbb4d9c21; Topsoil particle size distribution https://doi.org/10.5285/d6c3cc3c-a7b7-48b2-9e61-d07454639656; Headwater stream quality metrics, https://doi.org/10.5285/e305fa80-3d38-4576-beef-f6546fad5d45 ; Pond quality metrics, https://doi.org/10.5285/687b38d3-2278-41a0-9317-2c7595d6b882; Insect pollinator and flower data, https://doi.org/10.5285/3c8f4e46-bf6c-4ea1-9340-571fede26ee8; Bird counts, https://doi.org/10.5285/31da0a94-62be-47b3-b76e-4bdef3037360. [ABSTRACT FROM AUTHOR]

Published
2021
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5. Separation and identification of organic acid-coenzyme A thioesters using liquid chromatography/electrospray ionization-mass spectrometry

Author

Hobson Russell J, Joseph J. Dalluge, Frank Amore, Ravi R. Gokarn, Steven J. Gort, and Olga V. Selifonova

Subjects
chemistry.chemical_classification, Spectrometry, Mass, Electrospray Ionization, Electrospray, Chromatography, Chemistry, Elution, Coenzyme A, Electrospray ionization, Carboxylic Acids, Mass spectrometry, Biochemistry, Analytical Chemistry, chemistry.chemical_compound, Calibration, Escherichia coli, Mass spectrum, Molecule, Chromatography, High Pressure Liquid, Organic acid
Abstract

A method has been developed for the direct determination of coenzyme A (CoA) and organic acid-CoA thioesters in mixtures using directly combined liquid chromatography/electrospray ionization-mass spectrometry (LC/ESI-MS). Mixtures of CoA and organic acid-CoA thioesters were analyzed by LC/ESI-MS with detection of protonated molecular ions and characteristic fragment ions for each compound. The identities of the CoA-thioesters were established based on LC retention times and simultaneously recorded mass spectra. Monitoring of the CoA specific fragment ion at m/z 428 throughout the chromatogram provides a unique fingerprint for CoA content in the samples that corroborates the identification of organic acid-CoA thioesters in the mixtures. Furthermore, fragment ions arising from the ester linkage portion of the molecule allow unambiguous identification of the CoA esters in the samples. A second LC elution system was developed that allows the simultaneous separation and identification of 2-hydroxypropionyl-CoA (lactyl-CoA) and 3-hydroxypropionyl CoA (3HP-CoA), which have the same mass and identical MS fragmentation behavior. The utility of LC/ESI-MS employing this elution system is demonstrated by the determination of 3HP-CoA and lactyl-CoA (converted to CoA-thioesters from their corresponding free acids using CoA-transferase) in fermentation broths from Escherichia coli strains engineered for the production of 3-hydroxypropionic acid (3HP). External calibration employing a purified 3HP-CoA standard allowed indirect quantification of 3HP content in the broth with a precision of 1% (RSD). The feasibility of extending the method described above to perform LC/selected reaction monitoring-tandem mass spectrometry for direct determination of organic acid-CoA thioesters in cells was also demonstrated.

Published
2002

6. Glastir Monitoring & Evaluation Programme. First year annual report

Author

Williams, Bronwen, Emmett, Bridget, Abdalla, Mohamed, Anthony, Steven, August, Tom, Barrett, Gaynor, Biggs, Jeremy, Botham, Marc, Bradley, David, Brown, Mike, Carter, Heather, Chadwick, David, Cigna, Francesca, Cooper, David, Cooper, Jonathan, Cosby, Jack, Creer, Simon, Cross, Paul, Edwards, Francois, Edwards, Michael, Evans, Christopher, Ewald, Naomi, Fitton, Andrew, Garbutt, Angus, Grebby, Stephen, Greene, Sheila, Halfpenney, Ian, Hall, Jane, Harrison, Samantha, Harrower, Colin, Henrys, Peter, Hobson, Russell, Hughes, Steven, Isaac, Nick, Jackson, Bethanna, Jarvis, Susan, Jones, Davey, Keith, Aidan, Kelly, Martin, Korenko, J., Lallias, Delphine, Leaver, David, Lebron, Inma, Malcolm, Heath, Maskell, Lindsay, McDonald, J., Moxley, Janet, Norton, Lisa, O'Hare, Matthew, Owen, Anthea, Dos Santos Pereira, Gloria, Peyton, Jodey, Powney, Gary, Pywell, Richard, Rawlins, Barry, Robinson, David, Rorke, Stephanie, Rowland, Clare, Roy, David, Scarlett, Peter, Scholefield, Paul, Scott, Andy, Scott, Louise, Scott, Rod, Siriwardena, Gavin, Smart, Simon, Smith, Peter, Swetnam, Ruth, Taylor, Rachel, Tordoff, George, Van Breda, John, Vincent, Helen, Wagner, Markus, Waters, Emma, Watkins, John, White, J., Wood, Claire, Wright, Simon, Williams, Bronwen, Emmett, Bridget, Abdalla, Mohamed, Anthony, Steven, August, Tom, Barrett, Gaynor, Biggs, Jeremy, Botham, Marc, Bradley, David, Brown, Mike, Carter, Heather, Chadwick, David, Cigna, Francesca, Cooper, David, Cooper, Jonathan, Cosby, Jack, Creer, Simon, Cross, Paul, Edwards, Francois, Edwards, Michael, Evans, Christopher, Ewald, Naomi, Fitton, Andrew, Garbutt, Angus, Grebby, Stephen, Greene, Sheila, Halfpenney, Ian, Hall, Jane, Harrison, Samantha, Harrower, Colin, Henrys, Peter, Hobson, Russell, Hughes, Steven, Isaac, Nick, Jackson, Bethanna, Jarvis, Susan, Jones, Davey, Keith, Aidan, Kelly, Martin, Korenko, J., Lallias, Delphine, Leaver, David, Lebron, Inma, Malcolm, Heath, Maskell, Lindsay, McDonald, J., Moxley, Janet, Norton, Lisa, O'Hare, Matthew, Owen, Anthea, Dos Santos Pereira, Gloria, Peyton, Jodey, Powney, Gary, Pywell, Richard, Rawlins, Barry, Robinson, David, Rorke, Stephanie, Rowland, Clare, Roy, David, Scarlett, Peter, Scholefield, Paul, Scott, Andy, Scott, Louise, Scott, Rod, Siriwardena, Gavin, Smart, Simon, Smith, Peter, Swetnam, Ruth, Taylor, Rachel, Tordoff, George, Van Breda, John, Vincent, Helen, Wagner, Markus, Waters, Emma, Watkins, John, White, J., Wood, Claire, and Wright, Simon

Abstract

The Welsh Government has commissioned a comprehensive new ecosystem monitoring and evaluation programme to monitor the effects of Glastir, its new land management scheme, and to monitor progress towards a range of international biodiversity and environmental targets. A random sample of 1 km squares stratified by landcover types will be used both to monitor change at a national level in the wider countryside and to provide a backdrop against which intervention measures are assessed using a second sample of 1 km squares located in areas eligible for enhanced payments for advanced interventions. Modelling in the first year has forecast change based on current understanding, whilst a rolling national monitoring programme based on an ecosystem approach will provide an evidence-base for on-going, adaptive development of the scheme by Welsh Government. To our knowledge, this will constitute the largest and most in-depth ecosystem monitoring and evaluation programme of any member state of the European Union.

Published
2014

7. The exact transmission of texts in the first millennium BCE - an examination of the cuneiform evidence from Mesopotamia and the Torah scrolls from the western shore of the Dead Sea

Author

Hobson, Russell

Subjects
Bible, Dead Sea scrolls
Abstract

Certain ancient Near Eastern texts develop over time towards a reasonably stable state of transmission. However, the development towards a single ‘stabilised’ transmitted form that marks the biblical manuscripts between the second century B.C.E. and second century C.E. is often considered to permit the Hebrew bible a unique position in the ancient Near Eastern textual corpus. The degree to which the wider body of ancient Near Eastern texts actually support or undermine this position is the topic of this dissertation. The study begins by formulating a methodology for comparing the accuracy with which ancient texts of varying genres and languages were transmitted. Exemplars from the first millennium B.C.E. cuneiform evidence are selected for analysis on the basis of genre. Texts that are preserved in more than one ancient copy are compared to determine how much variation occurs between manuscripts of the same text. The study begins with representative texts from the Neo-Assyrian, Neo-Babylonian and Late Babylonian periods that range in date from the late eighth century B.C.E. to the third century B.C.E. The study then turns to the Torah scrolls from the Dead Sea area that range in date from the third century B.C.E. to the second century C.E. The accuracy with which the cuneiform texts were transmitted is then compared with the biblical evidence. The study finds that the most stable texts surveyed are those containing ritual instructions. The mechanisms that may have led to the exact transmission of the Torah in the late Second Temple period are discussed in the conclusion.

Published
2009

9. Jeremiah 41 and the Ammonite Alliance.

Author

Hobson, Russell

Subjects
MURDER, PILGRIMS & pilgrimages, RELIGIOUS travel, BIBLICAL commentaries, BIBLICAL criticism
Abstract

The article discusses the murder of seventy Northern pilgrims by Ishmael b. Netaniah at Mizpah. The author talks about the possible factors that contributed to Ishmael's assault on Mizpah. He tackles the election of Gedaliah b. Ahikam as governor outside of the Davidic line and head of an administrative centre at Benjaminte Mizpah. He mentions that Gedaliah has restored the peaceful life of military forces and civilians in Judean province. He adds that the restoration has ended after Ishmael, Judahite with the support of the Ammonite king Baalis, has assassinated Gedaliah and about 80 pilgrims travelling towards Mizpah.

Published
2010
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10. 955. Utilising Viral Gene Transfer to Model Polyglutamine Diseases and Study the Effects of Heat Shock Proteins

Author

Howarth, Joanna L., Kelly, Stephen, Glover, Colin P.J., Cheetham, Michael E., Bienemann, Alison S., Heywood, Darren J., Hobson, Russell J., Gallo, Jean-Marc, and Uney, James B.

Subjects
*GENETIC transformation
Abstract

An abstract of the article "Utilising Viral Gene Transfer to Model Polyglutamine Diseases and Study the Effects of Heat Shock Proteins," by Joanna L. Howarth and colleagues is presented.

Published
2005
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