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Posts Tagged ‘Clostridium difficile’

Clinell Manufacturers Choose A35 Workstation

We recently installed an A35 Anaerobic Workstation at GAMA Healthcare’s new state of the art
facility in Halifax, West Yorkshire. The company are best known for manufacturing Clinell
disinfectant wipes; one of the largest wet wipe suppliers to the NHS in the UK, and also exported to
over 50 countries worldwide.

Along with a growing number of our Anaerobic Workstation users, GAMA Healthcare is seeking to
cultivate Clostridium difficile – a harmful bacterium which infects the bowel and causes diarrhoea.
Growing these bacteria will allow the team to test the efficiency of the cleaning regime offered by
the Clinell wipes.

Pictured below with their new A35 are Mark Hallinan, Natalie Maguire and Evelina Cesnauskyte.

GAMA Healthcare Install photo

 

 

 

 

 

 

 

 

Nottingham iGEM Team Win Gold Medal!

Words and pictures provided by the University of Nottingham:

iGEM Team

The University of Nottingham’s 2018 iGEM team have been awarded a prestigious Gold Medal and were nominated for ‘Best New Composite Part’ at the recent International Genetically Engineered Machine (iGEM) competition in Boston, USA.

It is the first team from Nottingham to win a Gold Medal, which was awarded at the iGEM Jamboree event in Boston on Sunday 28th of October 2018.

iGEM

The iGEM competition began in January 2003 and currently attracts over 400 teams, from more than 45 countries and annually exceeds 5000 global undergraduate and post-graduate student participants. Teams use the principles of synthetic biology, the “Engineering of Biology”, to design biological parts, devices or systems to address a real-world problem or to perform a novel, previously unseen function. The best ‘parts’ of every project are then submitted in the form of a ‘BioBrick’ to the iGEM BioBrick registry for use by others.

University of Nottingham's iGEM team

University of Nottingham’s iGEM team

The Team

Nottingham’s team was composed of ten undergraduate students drawn from the Schools of Life Sciences, Biosciences, Computer Science, Mathematics and Social Sciences. For the duration of their project they were embedded within BBSRC/EPSRC Synthetic Biology Research Centre (SBRC) at Nottingham, under the overall guidance of Nigel P Minton and Philippe Soucaille and under the close supervision of a dedicated multidisciplinary team comprising Louise Dynes, Daphne Groothuis, Dr Christopher Humphreys, Dr Carmen McLeod, Dr Michaella Whittle and Dr Craig Woods.

Clostridium dTox

The team’s innovative synthetic biology project, Clostridium dTox; it’s not so difficile, aimed to develop a novel therapy for the treatment of disease caused by the superbug Clostridium difficile, colloquially known as C Diff.

C Diff infection is the most common cause of antibiotic-associated diarrhoea in the Western World and is a big problem in hospitals and healthcare-facilities. The disease symptoms are caused by the release of two major toxins, TcdA and TcdB by the bacterium. Under normal circumstances, a healthy gut microbiota prevents the proliferation of C Diff. However, when these good bacteria are obliterated by the use of broad-spectrum antibiotics, C. difficile proliferates and causes disease. One way to counter the expansion in numbers of toxin producing C Diff is to use competing strains that are not producing toxin as a probiotic.

The project’s aim was to engineer a C. difficile bacteriophage to produce factors that would suppress toxin production. The strategy adopted was to repress expression of both toxin genes (tcdA and tcdB) by targeting their mRNA using either antisense RNA (asRNA) or CRISPR interference (CRISPRi) technology (dCas9). The ultimate goal is a C. difficile-specific bacteriophage therapeutic which ablates toxin production in those cells that are infected with phage, converting them into health promoting probiotics. Unlike antibiotics, phage cause no collateral damage to the native gut microbiome.

The team was also nominated for ‘Best New Composite Part’. A composite part is a functional unit of DNA consisting of two or more basic parts assembled together. These must include all characterisation information and be added to the Registry.

Human Practices and Public Engagement

The team devised a number of activities that explored how Clostridium dTox could impact society. This included mining and carrying out a sentiment analysis of data from hundreds of social media comments on an online phage therapy video and exploring the current legislation surrounding phage therapy. They also researched what makes C. difficile such an important issue to society and how their project can help make a positive impact on communities by working towards the development of a novel therapy for its treatment. Finally they held a discussion group with non-scientists, and interviewed five leading scientific experts in the field, including the UK Public Health England lead on C. difficile infection, to understand how the team could make their project as effective as possible.

Public engagement was an important focus for the team, which developed hands-on workshops to communicate the project in local schools, libraries and to staff and students at the University. The team members have also been busy promoting their project via a range of social media platforms as well as by publishing articles in a local newspaper and in the University of Nottingham’s Impact Magazine.

Gold Medal

All of these aspects enabled the team to produce a project of high enough quality to win a Gold Medal at the Giant Jamboree, recognising the fulfilment of all the competition criteria. The Gold winning team members were; Lucy Allen, Hassan Al-ubeidi, Ruth Bentley, Sofya Berestova, Eun Cho, Lukas Hoen, Daniel Partridge, Varun Lobo, Fatima Taha and Nemira Zilinskaite.

“This was a tremendous achievement considering the short time that the team had to design, build and test the parts needed for the innovative project they devised. We broke new ground for iGEM by engineering a strict anaerobic bacterium, rather than the more traditional chassis other teams focus on. This was made possible by the extensive skills and expertise available through the involvement of SBRC researchers who gave so much of their free time to supervise the team”. – Nigel P Minton, SBRC Director, Nottingham

“Doing iGEM has given me a holistic understanding of the synthetic biology process. Coming from a Computer Science background, I had no knowledge of the science prior to iGEM, but working alongside talented team-mates meant that I left with a much better understanding of our project. I feel that my communication skills have improved since starting iGEM, as it has allowed me to interact with students and experts from many disciplines”. – Hassan Al-ubeidi, UG Computer Science.

“iGEM was an exciting challenge. As the sole modeller for our team, I improved my ability to work independently to research and solve problems. I learnt how to communicate my work in a way such that those with less technical knowledge can understand. Attending the Jamboree and seeing other projects made me appreciate the power of synthetic biology to build a better world”. – Ruth Bentley, UG Mathematics.

What the Judges Said

“Great project, great wiki!! You just light up so many questions in my mind and actually this is the key of synthetic biology! Thank you so much for your effort and all hard work!”

“Super interesting idea to use temperate phages for this! …. You are clear on your achievements and reasoning throughout, which is super refreshing. Great effort!”

“Really terrific modelling efforts! I really liked how thoroughly your work was documented on your wiki; everything was very clear.”

“Overall the project idea was very innovative, and you have great characterization on your parts. Good job!”

“Very impressive! It is very inspiring that your project used phage therapy, RNA interference and the extended application of CRISPR/Cas technology.”

“Amazing job, I hope that you continue this project.”

Sponsors

Nottingham’s iGEM team was generously supported by the University of Nottingham’s Research Priority Area in Industrial Biotechnology, through grant funding from the Wellcome Trust, the Biotechnology and Biological Sciences Research Council (BBSRC) and the National Institute for Health Research (NIHR) via the Nottingham Digestive Diseases Centre, by generous cash donations from Don Whitley Scientific Ltd, LanzaTech and Seres Therapeutics and through in-kind support from Qiagen, Millipore Sigma, Promega, Eppendorf, New England Biolabs, LabFolder and Snapgene.

Collaborators

The team also wishes to acknowledge support provided by the following collaborators: Team Biomarvel Korea and the teams from Imperial College London and the University of Warwick.

sponsors for Nottingham iGEM

iGEM_Fatima

Second Interview with Nottingham iGEM Team

You may have read our previous article that explained how Don Whitley Scientific Limited became involved in sponsoring a team from The University of Nottingham that have entered The International Genetically Engineered Machine (iGEM) Foundation competition.

iGEM is an independent, non-profit organization dedicated to education and competition, the advancement of synthetic biology, and the development of an open community and collaboration. iGEM runs the iGEM Competition – an international team competition made up of predominantly undergraduate students interested in the field of synthetic biology.

We spoke to another member of the team – Fatima Taha (pictured left) – to find out more about what the competition means to her and how she believes it might help in her future career. Fatima is a 3rd year Human Genetics student and we asked her:

What role do you play in the iGEM team? I have 3 roles really: I’m part of the wet lab team, part of the fundraising team, and also team leader – so the person to go to if there are any problems or issues that require bringing to the attention of the supervisors.

Have you used the Whitley Workstation and what did you use it for? I use the Whitley Anaerobic Workstation all the time. We grow up Clostridium difficile and take time points, pick colonies, etc inside the workstation. The cabinet works really well for us.

What do you think you will get out of the competition personally?

I was taking a gap year and worked for a year in a laboratory at a hospital. I fell in love with the hands-on process in that lab and decided I really want to continue working in the research sector – in a clinical application or in academia.

It’s more than just a synthetic biology competition – there’s so much more to it. We are encouraged to collaborate with other teams and with other people generally. It’s about getting the word out there. I enjoy the whole exercise of looking to engage with people about synthetic biology; getting involved with schools on the subject.

I’m really enjoying the communications side of the whole project and have written four articles for a university website and student magazine. The multi-disciplinary teams involved have such different strengths and weaknesses but it all comes together – and that’s fascinating. Going forward I am sure this whole process will have enhanced my own skill set.

What are you most looking forward to about going to Boston?

Meeting the other teams and seeing what they are doing. We have been working on our project for so long now, I’m excited to see what the others have been doing.

Don Whitley Scientific is proud to help sponsor some of the students to attend the finale in Boston, which we hope will provide the additional networking, team-building and general interaction to help the students in their future careers.  We wish Fatima and the team the very best of luck in the competition and hope that The University of Nottingham team brings home the grand prize.

More About iGEM

iGEMers state that they are building a better world by solving problems with the help of synthetic biology. The iGEM Competition inspires nearly 6,000 students each year to work in teams to address unique challenges in their local communities.

They celebrate team achievements at the annual Giant Jamboree (24-28 October) by showcasing projects from participating teams and awarding medals, prizes, and the grand prize, the BioBrick trophies.

Their aim is to inspire responsible innovation through efforts in biosafety, biosecurity and public outreach.

iGEM Community

The iGEM community is made up of international trailblazers from over 45 countries around the world.

In 2017 iGEM launched the After iGEM program. This program supports over 30,000 iGEMers – students and instructors – who have gone through the competition since its inception in 2004. This global network is leading the field, taking what they learned in the competition and expanding it to continue to build a better world.

 

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Dan Partridge, iGEM participant

DWS Supports Nottingham iGem Team

Don Whitley Scientific Limited were recently approached by Professor Nigel Minton from the University of Nottingham requesting some sponsorship to enter a global competition. The International Genetically Engineered Machine (iGEM) Foundation is an independent, non-profit organization dedicated to education and competition, the advancement of synthetic biology, and the development of an open, collaborative community. iGEM runs the iGEM Competition – an international team competition made up of predominantly undergraduate students interested in the field of synthetic biology.

We spoke to one of the team – Daniel Partridge (pictured below) – to find out more about the competition and what it means to him personally. Dan is a 3rd year BSc student studying Biotechnology. We asked him:

Can you explain the project you are working on? Our project centres on the Clostridium difficile bacteriophage. With the increase in antibiotic resistance, we need to develop a more precise method to attack the pathogen C. difficile, as this bacterium can cause unpleasant and sometimes serious bowel problems.

Do you play a specific role in the iGEM team? There are 10 students and five supervisors in the team. We started out with specific roles. There are three lab teams: two are trying to reduce toxin production of C.difficile using genetic engineering techniques – RNA interference and CRISPR dCas9 – and the third group is the promoter team (contribution). The non-lab teams look at human/outreach, financing, computing and modelling.

How many teams are in the competition? There are about 400 teams across the world.

What do you think you will get out of the competition personally? I am going into my 3rd year so the opportunities this competition will provide to get into problem solving and thinking-on-the-spot will be invaluable in my future career. With the sheer number of scientists that will be in Boston, it will be a fantastic networking opportunity. I understand that we may also be given the chance to look round the laboratories of some organisations in the area.iGEM team photo

Don Whitley Scientific recognises the importance of initiatives like this that strive to further scientific breakthroughs and we are happy to support the team. We wish Dan and the team the very best of luck in the competition and hope that their project brings home the grand prize. Click here to learn more about SBRC Nottingham.

More About iGEM

iGEMers state that they are building a better world by solving problems with the help of synthetic biology. The iGEM Competition inspires nearly 6,000 students each year to work in teams to address unique challenges in their local communities.

They celebrate team achievements at the annual Giant Jamboree by showcasing projects from participating teams and awarding medals, prizes, and the grand prize, the BioBrick trophies.

Their aim is to inspire responsible innovation through efforts in biosafety, biosecurity and public outreach.

iGEM Community

The iGEM community is made up of international trailblazers from over 45 countries around the world.

In 2017 iGEM launched the After iGEM program. This program supports over 30,000 iGEMers – students and instructors – who have gone through the competition since its inception in 2004. This global network is leading the field, taking what they learned in the competition and expanding it to continue to build a better world.

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Professor Tom Riley Lab

30 Year Commitment for Professor Tom Riley

Professor Tom Riley from Pathwest Laboratory Medicine and The University of Western Australia has been a committed Don Whitley Scientific customer for over 30 years. In the past few years he has installed two A35 Anaerobic Workstations into his lab to replace a Whitley MK III Workstation that had given him over 20 years of reliable performance.

Professor Riley started using anaerobic workstations over 30 years ago when the capacity and time consuming operation of jar gassing systems was deemed unsuitable for the number of samples he had. Tom explained to Don Whitley Scientific how he came to use Whitley Workstations. “My original anaerobic chamber from a manufacturer in Australia (that doesn’t exist anymore) was hopeless.”. The next option for Professor Riley was a Whitley MKIII Anaerobic Workstation, which served his lab for over 20 years, and was the first Whitley Anaerobic Workstation in Australia. His two Whitley A35 Anaerobic Workstations currently play a part in work that Professor Riley explains as “almost exclusively research into Clostridium difficile. This includes everything from diagnostics to pathogenesis and epidemiology”.

Although the MK III provided a reliable anaerobic environment for such a long time in Professor Riley’s lab, there are new benefits provided by the A35 Anaerobic Workstation that Tom finds particularly useful, such as the Instant Access Porthole System and Letterbox entry. The Instant Access Porthole System is unique to DWS, allowing entry to the chamber in seconds without the need of gloves or sleeves. Letterbox entry allows the user to introduce Petri dishes and similar small items into the anaerobic workstation environment in no time at all. As aforementioned, Tom also needed extra capacity to process a large number of samples. Compared to using the MKIII or anaerobic jars, the main chamber of an A35 Anaerobic Workstation will accommodate between 400-600 x 90mm Petri dishes depending on whether plate carriers are used, which accessories and system options have been incorporated and how much working space is required.

The space and reliability provided by Whitley Workstations has served Professor Tom Riley well for over three decades now. He is part of a long list of satisfied Whitley Workstation users around the world, who are all doing fantastic work with Don Whitley Scientific products.


 

Recent papers from the Riley lab are listed below

 

 

Biomedical Scientist_Clinical Microbiology of Anaerobes Course

Campylobactor Research with Microaerobic Workstations

The following words were provided by Microbiology International 

Cultivation of microaerophilic organisms from environmental niches such as Campylobacter species and Helicobacter pylori requires precise calibration of oxygen levels and other growth parameters. Microbiology International, North American distributor for Don Whitley Scientific, have been installing anaerobic and microaerobic incubation chambers for more than 20 years, and would like to introduce some of the research being carried out around the world. Click here to view our Microaerobic Workstations.

The complex interactions of microbial communities populating the human gastrointestinal tract with their host and with invading pathogens are paramount to safeguarding not only a healthy gut but also our general health. Microaerophilic bacterial species, such as Campylobacter jejuni, can cause gastro-enteric infections due to their ability to survive and grow in lower oxygen environments, which they encounter in the human gastro-intestinal tract. At the Ottawa Institute of Systems Biology, Dr. Alain Stintzi researches topics as diverse as iron homeostasis and oxidative stress and the competitive advantage of metabolizing L-Fucose in Campylobacter jejuni. Using a Don Whitley Scientific microaerobic workstation, Dr. Stinzi states that “One objective of our research is to understand how enteric pathogens such as Campylobacter jejuni acquire essential nutrients, adapt to the harsh conditions of the intestine and interact with the host’s microbiota to cause disease.”

At Ohio State University, Dr. Jeffrey Lejeune’s research is focused on prevention of diseases caused by food-borne pathogens in plants and animals, including C. jejuniE. coli O157, and Clostridium difficile. His work on antibiotic resistance in Campylobacter and other enteric pathogens has prompted the UN’s Food and Agriculture Organisation to recruit him to provide technical support and guidance in an international group working on antibiotic resistance. Don Whitley Scientific anaerobic and microaerobic workstations , sold in the US by Microbiology International, provide precise gas control for a sustainable low oxygen environment. Up to four gasses can be combined to create the ideal atmosphere for fastidious microorganisms.

In the UK, Dr. Andrew Grant at the University of Cambridge is investigating the Campylobacter jejuni secretome and diarrhoeal disease in a gnotobiotic piglet model, using the MACS VA500 and more recently, the M95 Microaerobic Workstation. His work to elucidate host-pathogen interaction and virulence strategies will yield new options for therapy and vaccination. Don Whitley Scientific will be there for him, and our other researchers, every step of the way.

 

From: Stahl et al. (2011) “L-fucose utilization provides Campylobacter jejuni with a competitive advantage“ Proc Natl Acad Sci U S A. 2011 Apr 26;108(17):7194-9

From: Stahl et al. (2011) “L-fucose utilization provides Campylobacter jejuni with a competitive advantage“ Proc Natl Acad Sci U S A. 2011 Apr 26;108(17):7194-9

From: Tang et al. (2017) “Rising fluoroquinolone resistance in Campylobacter isolated from feedlot cattle in the United States” Scientific Reports 7: 494

From: Tang et al. (2017) “Rising fluoroquinolone resistance in Campylobacter isolated from feedlot cattle in the United States” Scientific Reports 7: 494

 

 

 

 

 

 

 

 

 

 

 

 

 

 

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Why Choose a Whitley Workstation?

There are many reasons to choose a Whitley Workstation when it comes to Anaerobic, Hypoxic or Microaerophilic work. We can discuss these with you anytime, but we also have plenty of satisfied customers who have expressed why using a Whitley Workstation improves their working methods and results.

Over the years, customers have supplied us with many testimonials about their Don Whitley Scientific products. From these we can see that not only have Whitley Workstations become approved by fantastic researchers worldwide, but we can also help promote the amazing work that is done by our customers.

Dr Vaibhao Janbandhu at the Victor Chang Cardiac Research Institute (VCCR) in Sydney, Australia uses a Whitley H35 Hypoxystation in his work on finding new ways to stimulate heart regeneration during ageing and after heart attack. He uses his H35 Hypoxystation to isolate, culture and characterise adult cardiac stem cells. In Dr Janbandhu’s words the H35 is “an integral part of the project to advance the project aims”.

In this video testimonial, Jane Freeman at Leeds General Infirmary explains how her Whitley A95 Workstation improves the working methods in her Clostridium difficile research. Jane reports that she and her team are able to use the workstation for “several hours at a time in relative comfort” and that the workstation is able to house all the technical equipment her team requires. This allows “the whole experiment to be performed in optimum conditions without introducing air at all”. Jane explains that “reliability, versatility and space are the significant benefits of the workstations in our work on Clostridium difficile“.

The Institute of Cancer Research in London is one of the world’s most influential research institutes, with an outstanding record of achievement dating back more than 100 years. At the Institute, George Poulogiannis uses a combination of Whitley i2 Instrument Workstation (with Seahorse XF Analyzer) and Whitley H35 Hypoxystation in his research into breast cancer. Hypoxia is a key factor in the “Hallmarks of Cancer” and this team are studying the role of hypoxia in cell invasion and metastasis, oncogene-induced senescence and resistance to current treatment options. The i2 and H35 replicate a physiologically relevant atmosphere for these studies, enabling consistent and reliable results. This combination of Workstations is also used by Dr Ayse Latif, who is researching gynaecological cancers at The University of Manchester.

Don Whitley Scientific would like to take this opportunity to thank all customers who have provided testimonials. If you would be interested in supplying a testimonial, please contact Alex_Rhodes@dwscientific.co.uk.

Take a look at our other testimonials

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Taking on the C difficile challenge

In the United States, nearly half a million infections are caused by Clostridium difficile (C. difficile) annually, with approximately 17% experiencing at least one recurrence; almost 6% of patients die within 30 days of diagnosis.

The standard first-line treatment for C. difficile infection (CDI) relies on the antibiotic metronidazole; however, metronidazole is not as effective for severe cases of CDI, due to its rapid absorption in the upper GI tract. At Texas A&M, Julian Hurdle’s group are using Whitley Anaerobic Workstations to improve treatment outcome by developing modified derivatives of metronidazole.

Julian Hurdle and Philip Cherian describe their research in their 2015 paper “Gastrointestinal localization of metronidazole by a lactobacilli-inspired tetramic acid motif improves treatment outcomes in the hamster model of Clostridium difficile infection“. In essence, the group synthesized a series of metronidazole derivatives with a tetramic acid motif utilized by Lactobacillus strains, assaying their efficacy in C. difficile cultures growing in a Whitley A35 Workstation. In animal experiments, the modified compounds were found to exhibit significantly better efficacy in treating CDI, due to minimal absorption as compared to the unmodified drug. The A35 Anaerobic Workstation enables comfortable gloveless access to the chamber, where cultures are manipulated and incubated under consistent anaerobic conditions. Features such as HEPA containment and anaerobic conditions monitoring system guarantee that the atmosphere inside the workstation is absolutely anaerobic and particulate free. Drs Hurdle and Cherian have recently applied for a patent for compounds and methods based on their C. difficile research.

All over the world, labs are using Whitley Workstations to research C. difficile. In the UK, C. difficile is still a challenge with 17,925 cases reported in 2015. Leeds General Infirmary now has five Whitley Workstations. Dr Jane Freeman at Leeds uses an A95 Anaerobic Workstation, our largest workstation, which can accommodate two technicians simultaneously and has a capacity of up to 1400 plates. The team at Leeds appreciate the reliable anaerobic atmosphere, the spacious working area, and the ability to keep instrumentation inside the workstation. Watch this YouTube video highlighting  Dr Freeman’s work with C. difficile.

 


By Dr Burga Kalz Fuller

Microbiology International is the exclusive distributor for Whitley Anaerobic Workstations in North America.

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New Video Testimonial – Optimal conditions for Clostridium difficile

Dr Jane Freeman, from Leeds General Infirmary and The University of Leeds, spoke to Don Whitley Scientific about her work with the Whitley A95 Anaerobic Workstation. 

Dr Freeman is part of the Healthcare Associated Infections Research Team that works at Leeds General Infirmary. Jane’s work involves investigating Clostridium difficile, a bacteria that can infect the bowel, causing diarrhoea and sometimes even more serious bowel conditions. Jane uses a Whitley A95 Anaerobic Workstation in her work and  she also uses an in vitro gut model (as shown in the video) to mimic the effects of C difficle.

The Whitley A95 Anaerobic Workstation offers a spacious work area with four portholes and a large capacity airlock. Dr Freeman states this large working area enables her to use all the equipment she needs without taking her samples out of the anaerobic conditions. The team at Leeds General Infirmary have five Whitley Workstations, as they provide a reliable anaerobic atmosphere in which to incubate and manipulate samples.

This video highlights how the Whitley A95 Anaerobic Workstation provides reliability, versatility and space for this important work with Clostridium difficile.

Clostridium difficile studies can be done in a Whitley Workstation

Whitley Workstation used in Clostridium difficile study

The Whitley A35 Workstation has recently been used by a group at Monash University in a study relating to toxins affected by Clostridium difficile.

This very interesting research paper, entitled “CdtR Regulates TcdA and TcdB Production in Clostridium difficile“, was written by researchers at Monash University in Melbourne, Australia. The results from the study “establish CdtR as an important virulence regulator in two clinically important, epidemic strains of C. difficile, and further highlights the need to investigate regulatory mechanisms of important virulence factors in diverse strain backgrounds.”  The paper proves for the first time that “TcdA and TcdB production is linked to the production of CDT by a common regulatory mechanism”.

Click here to read the full paper