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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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Workstation Installation at Trinity College Dublin

Don Whitley Scientific and our distributor in Ireland, Davidson and Hardy, recently installed equipment at Trinity College Dublin to help with research into cell metabolism.

Stephen Maher, Assistant Professor at Trinity College Dublin first experienced using a Whitley Workstation when working with a group at The University of Hull.  In his Trinity College Dublin lab, Don Whitley Scientific recently installed a Whitley H35 Hypoxystation, to be used connected to a Whitley i2 Instrument Workstation. The combination of these two units should help Stephen Maher greatly in his research.

The Whitley H35 Hypoxystation is ideal for cell and tissue culture researchers who want to accurately control oxygen, carbon dioxide, temperature and humidity. A removable front allows large quantities of samples or pieces of equipment to be placed inside the unit for use within these specific, physiologically relevant conditions. The i2 Workstation was developed in response to a rising number of enquiries from scientists wanting to use Seahorse Extracellular Flux (XF) Analyzers in hypoxic conditions and were dissatisfied with the solutions available. This workstation can be used as a stand-alone unit or connected to a Whitley Hypoxystation via the new Whitley Transfer Tunnel, enabling preparation of cell lines under hypoxic conditions and their transfer directly into the i2 without exposure to air.

 

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Explore the Whitley Hypoxystation Range

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Molecular Targeting of Hypoxia in Radiotherapy

Most solid tumours exhibit areas of both chronic and acute hypoxia, all of them evolving dynamically as a function of cellular growth, vascularisation, oxygen consuming metabolism and therapy response. Tumour hypoxia, generally far below 1% oxygen, correlates with increased recurrence rates and decreased survival rates in most cancers, so the recent review by Hypoxystation users Rey et al. describing “Molecular Targeting of Hypoxia in Radiotherapy” gives a valuable overview of the mechanisms cancer cells have developed to respond to hypoxia.

Dr. Rey of the Princess Margaret Cancer Centre in Toronto, Canada, and his co-authors Luana Schito, Marianne Koritzinsky and Brad Wouters have contributed vastly to our knowledge about the cellular response to hypoxia in the context of tumour behavior. Since 2009, they have acquired four Hypoxystations for their lab, in order to culture cells under conditions which authentically mimic the physiological environment of cancer. The Hypoxystation provides a closed workstation format for rigorous control of oxygen, CO2, temperature and humidity, facilitating accurate regulation of cell culture conditions as the in vivo tumour situation is simulated.

 

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In their 2016 review, Rey et al. describe the cellular response to the complex interplay of temporal and spatial variations in oxygen levels, and the rippling effects exerted on vascular, stromal and immunological responses.

 

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By Burga Kalz Fuller, HypOxygen

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Hypoxia and Tumor Metabolism in Whistler

Information on an upcoming exhibition in Whistler for HypOxygen. Words by Dr Burga Kalz Fuller.

At 2140 feet in Whistler, BC, the air will be getting thinner at the Keystone Symposia on “Adaptations to Hypoxia in Physiology and Disease” joint with the meeting on “Tumour Metabolism: Mechanisms and Targets”. But after all, hypoxia is what we do at HypOxygen, so we are very excited to be spending time at altitude with old friends and new ones on March 5-9.

At the Keystone Symposia in Whistler, HypOxygen will be exhibiting Whitley Hypoxystations for low oxygen cell culture under in vivo conditions. Conceived as an incubator workstation, but allowing gloveless access “to avoid spikes of normoxia” for cancer cells accustomed to very low oxygen, the Hypoxystation enables researchers to culture and manipulate cells growing at consistent oxygen, CO2, humidity and temperature. Another member of the Hypoxystation family, the i2 Instrument Workstation, was developed specifically to house instrumentation such as the Agilent Seahorse XF Analyzer for metabolism assays at hypoxia.

Since seeing is believing, we are greatly looking forward to talks and posters by a number of researchers who use Hypoxystations for their hypoxic cell culture. The broad range of these researchers’ presentations clearly illustrates how closely oxygen availability is linked to cancer cell behavior and metabolism, as the Hallmarks of Cancer are influenced and even determined by hypoxia in the tumour environment. These Hypoxystation users will be presenting data in Whistler:

  • Nicholas Denko, Ohio State University, USA
    Hypoxic Regulation of Mitochondrial Function
  • Almut Schulze, University of Würzburg/Theodor-Boveri Institute, Germany
    Targeting Glucose and Lipid Metabolism in Cancer
  • Janine T. Erler, University of Copenhagen, Denmark
    Hypoxia-Driven ECM Remodelling during Cancer Progression
  • Navdeep S. Chandel, Northwestern University, USA
    Why Mammalian Cells Respire?
  • Sara M. Timpano, University of Guelph, Canada
    Human Cells Cultured Under Physiological Oxygen Utilize a Different Mode of Translation Initiation, Have Higher Proliferation Rates, Less Oxidized DNA and More Tubular Mitochondria
  • Karen H. Vousden, Beatson Institute for Cancer Research, UK
    A Role for p53 in the Adaptation to Metabolic Stress
  • Cormac Taylor, University College Dublin, Ireland
    The Role of Hypoxia in Immunity and Inflammation
  • Eyal Gottlieb, Technion Integrated Cancer Center, Israel
    Metabolic Dependencies of Leukemic Stem Cells
  • Bradly G. Wouters, University Health Network, Canada
    ULK1 Regulates Oxygen Metabolism, Hypoxia Tolerance and Is a Therapeutic Target in Pancreatic Cancer
  • Ester M. Hammond, University of Oxford, UK
    Ribonucleotide Reductase Favors the RRM2B Subunit to Maintain DNA Replication in Hypoxia

 

Please stop by our exhibit at the Whistler Conference Center to learn more about the ways the Hypoxystation can recreate the tumour environment for your cancer research. We also have a “heart-warming” gift for you!

 

Hallmarks of Cancer

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Hypoxia and the Hallmarks of Cancer

Therapeutic Targeting of Hypoxia and HIFs in Cancer. Dr Burga Kalz Fuller from US distributor HypOxygen has summarised this study that outlines the Hallmarks of Cancer.

“Tumour hypoxia and HIFs affect most of the cancer hallmarks… and contribute to chemo- and radiotherapy resistance.” In their review from 2016, Wigerup, Pahlman and Bexell of Lund University in Sweden discuss how hypoxia inducible factors HIFs regulate the hypoxic microenvironment in cancer, and the therapeutic strategies that are being developed to improve patients’ prognosis. Dr. Sven Pahlman’s lab has been using the H35 Hypoxystation for more than 5 years, to research SCLC and neuroblastoma, and their data is contributing to the understanding of the role of oxygen levels in the progression of cancer.

Hypoxia and HIF-1α and 2α expression in cancer usually signify a worse prognosis, but most hypoxia-induced transcriptional, translational, and epigenetic changes are cell-type specific. Many effects engendered by hypoxia are mediated directly or indirectly via HIF pathways, and most are causative of the iconic “Hallmarks of Cancer” that Hanahan and Weinberg introduced in 2000 and expanded in 2011. Hypoxia induces increased autophagy, apoptosis, and aberrant cell proliferation; neoangiogenesis mediated by VEGF and PDGF-β; proliferation of cancer stem cells; metabolic reprogramming to satisfy energy and synthetic requirements in proliferating cells; modulation of inflammation and immune responses; genomic instability through increased mutagenesis and diminished DNA repair; and metastasis as hypoxia induces epithelial-to-mesenchymal transition and degradation of the extracellular matrix. Assaying the relationship between hypoxia and the Hallmarks of Cancer benefits significantly from the physiological atmosphere mimicked in the Hypoxystation, a closed-culture hypoxia workstation controlling gasses, temperature and humidity.


Visit Don Whitley Scientific and HypOxygen at

Keystone Adaptations to Hypoxia and Tumour Metabolism

Location: Whistler, BC  Date: 5th – 9th March

Sven Pahlman


In their review, Wigerup and Pahlman describe the role tumour hypoxia plays for cancer therapy and treatment resistance, as oxygen levels, production of reactive oxygen species ROS, and HIF activity are intertwined actors in the cancer battle. Any and all effects of hypoxia are cell-type specific; however, numerous studies indicate that HIF’s mediate chemoresistance, suggesting that HIF-1 and 2 inhibitors can effectively support cancer therapy. The authors state that “since hypoxia is a hallmark of solid tumours and mediates aggressive, metastatic, and resistant disease, it is arguably one of the most attractive therapeutic targets in cancer.” Strategies selectively targeting hypoxia for cancer therapy include hypoxia-activated prodrugs; inhibitors of HIF mRNA and protein expression; and inhibitors of downstream HIF signalling pathways such as VEGF. Effective drug research relies on authentic replication of the hypoxic environment for cell culture: the Hypoxystation used in the Pahlman lab is able to accommodate long-term assays with sterile steam humidification and HEPA clean air. The Hypoxystation concept “Choose your Atmosphere – Define your Environment” is the best way to ensure cell culture reflects physiology in cancer research and therapy.

Hypoxia is at the heart of the Hallmarks of Cancer, and results such as these from the Pahlman lab make the cancer research community hopeful that “HIF inhibition is likely to be a powerful therapeutic approach” to eradicate cancer.

Hallmarks of Cancer

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MEDLAB 2017

Export representatives from Don Whitley Scientific have been at MEDLAB 2017, an international conference in Dubai that is one of the biggest exhibitions for the Middle East healthcare industry. Formerly the event was a joint venture with Arab Health, but MEDLAB is now its own independent event. Don Whitley Scientific had exhibition stand at the event which displayed a Whitley A35 Anaerobic Workstation and the Whitley Jar Gassing System.

MEDLAB houses 700 exhibitors from 38 countries, giving delegates access to cutting edge products and services from all continents. As well as this huge exhibition, the event also features an extensive lecture programme including topics such as Haematology, Clinical Microbiology and Immunology. The event was held at the Dubai International Convention and Exhibition Centre. As well as offering delegates access to new and developing products from a diverse range of companies, the event was also an opportunity for those wanting to make business connections and expand their distributor network.

 

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Joe and Alun at the Don Whitley Scientific exhibition stand

Visitors to the Don Whitley Scientific got up close with two key DWS products. Firstly, the Whitley Jar Gassing System in which users can create perfect conditions for growing anaerobes in jars in just 2 minutes, a popular product which has been manufactured and sold by Don Whitley Scientific for decades. The Whitley A35 Anaerobic Workstation shares a similar concept to the WJGS in creating a reliable, anaerobic environment. The A35 provides the ability to manipulate samples in a sustainable environment where parameters can be altered to create the required conditions, unique features such as the Instant Access Porthole system make working in the chamber simple and straight forward.

MEDLAB is always a popular event and one that Don Whitley Scientific’s export team ensure to attend every year to promote the best Anaerobic Workstations on the market.

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Don Whitley Visits The Francis Crick Institute

Dr Don Whitley, chairman and founder of DWS, recently visited The Francis Crick Institute in London to see the recently installed Whitley H45 Hypoxystation at the site. 

The Crick has moved into a brand new state-of-the-art building in the centre of London. Situated next to Kings Cross and St Pancras stations, The Crick brings together 1500 scientists and staff working collaboratively in the biggest biomedical research facility under one roof in Europe. The work at The Crick covers many disciplines and applications in biomedical research, all with the aim of improving understanding of human health and disease.

 

Don Whitley established Don Whitley Scientific in 1976 and today Don Whitley Scientific Limited is a leading international supplier of innovative equipment and services to the microbiology and tissue culture industries. Recently DWS installed a H45 Hypoxystation into the institute, and Don Whitley went to visit the new customer with a member of the sales team.

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Don Whitley (far right) with one of the groups that will be using the Whitley H45 Hypoxystation

The Whitley H45 Hypoxystation has sufficient space to accommodate a variety of equipment whilst still providing generous working and incubation areas. Whitley Hypoxystations can be equipped with a range of unique options and features, including CO2 monitoring and automatic dehumidification fitted as standard, features that will make working with the H45 easy and efficient for the team at The Crick.

With his name featured on products in hundreds of clinical and research laboratories worldwide, it can be said that the staff at The Crick were excited to meet Don Whitley himself. The excitement was shared by Don, who enjoyed looking around one of the most exciting centres for biomedical research in the UK and taking a few pictures along the way.

 

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Don takes a photo looking out over the atrium of The Francis Crick Institute

 

Dr John Heap, Imperial College London

Realising the potential for synthetic biology

At Don Whitley Scientific we love to hear about the different applications and scientific disciplines in which our workstations are used. We recently came across a video of a presentation at the World Economic Forum in 2015. The clip is entitled: “Hacking exotic organisms and putting them to work“. Dr John Heap, who is an academic scientist and research group leader at Imperial College London, talks about harnessing diverse organisms including anaerobic bacteria, and realising the potential of synthetic biology for a variety of applications.

You will also see a very quick shot of Dr Heap using his Whitley Workstation!

For more information about Dr Heap and his work, here’s a link to the Heap Lab website.

 

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Paper Highlights How Cells Respond to Stress Conditions

Dr Burga Kalz Fuller, Product Manager at HypOxygen (Don Whitley Scientific’s Hypoxystation provider in the USA/Canada) summarises a study entitled “Adaptation to Stressors by Systemic Protein Amyloidogenesis”. The paper, by Whitley Hypoxystation users Tim Audas and Stephen Lee, details how certain cells activate a process of systemic amyloidogenesis, which allows them to survive during difficult conditions.

Cells facing environmental threats have developed numerous coping mechanisms, and Hypoxystation users Tim Audas and Stephen Lee have uncovered a fascinating new cellular strategy to remain viable under stress and restore homeostasis when the crisis ends. In their recent paper “Adaptation to Stressors by Systemic Protein Amyloidogenesis“, they describe a physiological process of amyloidogenesis which cells activate under stress conditions, such as hypoxia and acidosis, to remove copious amounts of heterogeneous proteins from circulation, enabling cells to survive in a dormant state. This discovery expands our current view of amyloids as a rare and pathological phenomenon associated with neuropathies such as Alzheimer’s and Parkinson’s diseases, and exposes a novel post-translational, regulatory form of protein organization.

Using a combination of Congo red staining, proteinase K digestion, and OC antibody detection on cells exposed to a variety of stimuli in the Hypoxystation, Audas et al. were able to identify nuclear foci consisting of immobilized, insoluble protein in a crossed β-sheet conformation which they named A-Bodies. In amyloidogenic proteins such as VHL and RNF8, an ACM (amyloid-converting motif) containing arginine and histidine was identified as essential for capture specifically in the A-bodies; a similar motif was also identified in the pathological β-amyloid associated with Alzheimer’s disease. Upon environmental insult, the ACM interacts with ribosomal intergenic spacer RNA (rIGSRNA) to concentrate the proteins and trigger their polymerization in the A-bodies allowing the cells to enter a dormant state.

Audas et al. exerted this type of severe stress on the cells through incubation at pH 6.0 and 1% oxygen in the H35 Hypoxystation by Don Whitley Scientific. The Hypoxystation’s closed workstation format and rigorous control of oxygen, CO2, temperature and humidity facilitate accurate regulation of cell culture conditions as the in vivo situation of adverse environmental stimuli is simulated. Upon reversion to standard growth conditions (21% oxygen and pH 7.4), the A-bodies dissipated within 4 hours and protein was refolded into the native conformation. The hypoxic and acidotic conditions simulated in the Hypoxystation are also characteristic of the tumor microenvironment, where mouse xenograft assays identified the same process of rIGSRNA-mediated A-body formation causing cancer cell dormancy.

Read more on this paper by HypOxygen

Read the full paper here

WASP Touch

WASP Touch Training for Spanish Distributors

As part of the continued roll out of the Whitley WASP Touch to our distributors around the world, Steve Robertson and Joe Walton recently visited Spain. Here they carried out Whitley WASP Touch training with our distributor there. This included a short refresher course on spiral plating techniques and bespoke WASP Touch demonstrations. The training was followed by a series of customer visits.

 

Feedback on the Whitley WASP Touch continues to be positive. In particular, the intuitive touch screen and automatic filling of the wash station are seen to be big positives in customers’ eyes. This, coupled with the reputation for quality of the WASP 2 (predecessor to the Whitley WASP Touch), continues to position the new spiral plater as a top quality laboratory device.

 

The Whitley WASP Touch is designed to fit the needs of modern microbiology laboratories. It is fundamentally different from its predecessors, with no need for a separate vacuum source. An easy to use touch screen and automatic sanitising system (patent pending) ensures that set up and operation is simple and straight forward.

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