Death and Dysbiosis: Linking Neonatal Sepsis and Gut Microbes

Nearly half of global deaths in children under 5 are caused by neonatal sepsis and despite enhancements in knowledge of other fatal childhood illnesses, neonatal sepsis is still proving difficult to tackle [1]. Clinical practice is challenged by the fact that there is no universally accepted definition of neonatal sepsis, making surveillance, diagnosis and treatment more challenging [1]. Neonatal sepsis is characterised by infection of the bloodstream; there are 2 different types including early-onset sepsis (EOS), which occurs before 72 hours of life and late-onset sepsis (LOS), which occurs after 72 hours of life [2]. EOS is typically caused by transmission of pathogens such as group B Streptococcus, Escherichia coli and Listeria monocytogenes from the female genitourinary system to the newborn or foetus, whereas LOS is usually caused by transmission of pathogens such as coagulase-negative Staphylococci post-birth or during birth [2]. Since the 1990s, incidence of EOS has improved but LOS rates have generally not changed, encouraging scientists to improve their understanding of the infection and establish a possible cause for this [2].

Over the decades of research into neonatal sepsis, scientists have found that the gut microbiome plays a vital role in neonatal health [3]. Gut pathogen colonisation can be a precursor to many infections in neonates, so understanding gut dysbiosis and sepsis is paramount for preventative and therapeutic strategies [3]. Iqbal et al. delved into this area of research by investigating the difference in the gut microbiota of babies who developed sepsis compared to those who did not [3]. Faecal samples were collected on day 4 of life from 182 preterm neonates (91 who had sepsis and 91 without sepsis) and incubated at 37oC for 24 to 48 hours. For efficient growth of any anaerobes, the samples were inoculated on 5% sheep blood agar and neomycin blood agar with a metronidazole disc and incubated in a Whitley A35 Workstation [3]. For infants without sepsis, faecal samples were also taken on day 14 but the second sample for those with sepsis was taken when blood culture bottles showed growth of pathogenic bacteria [3]. Evidence shows that there is a difference in gut microbiome depending on whether the babies have sepsis or not as on day 14, there was a significant difference in E. coli with 64.7% cultured from those diagnosed with sepsis, compared to 50% in those without sepsis [3]. Similarly, there were much higher rates in those with sepsis for other bacteria including Bifidobacterium spp. and Klebsiella pneumoniae, compared to those without sepsis [3]. Among these findings, they also discovered that there were microbial differences depending on the mode of delivery; for vaginal births, there was a higher prevalence of beneficial gut flora bacteria such as Bifidobacterium spp. and Enterococcus faecalis compared to caesarean section births [3].

Understanding the link between neonatal dysbiosis and sepsis to a deeper level will prove beneficial for successful treatment, so this kind of research is valuable as it will hopefully lead to a lower mortality rate of neonatal sepsis.

Written by DWS Microbiologist, Kirsty McTear

Read the paper by Iqbal et al. on our free published paper database

References 

1. Strunk T, J Molloy E, Mishra A, Bhutta ZA. Neonatal bacterial sepsis. The Lancet. 2024 May 26;404(10449):277–93.

2. Singh M, Alsaleem M, Gray CP. Neonatal Sepsis. [Updated 2022 Sep 29]. In: StatPearls [Internet]. Treasure Island (FL): StatPearls Publishing; 2024 Jan-. Available from: https://www.ncbi.nlm.nih.gov/books/NBK531478/

3. Iqbal F, Shenoy PA, Siva N, Vandana KE, Purkayastha J, S. Lewis LE. From dysbiosis to disease: Tracing gut microbiota’s role in neonatal sepsis. Clinical Epidemiology and Global Health. 2024 Oct 8;30:101809.

The Silent Pandemic: Can We Use Antimicrobial Peptides to Beat Antimicrobial Resistance?

European Antibiotics Awareness Day (EAAD) occurs every year on 18 November. Established in 2008 by the European Centre for Disease Prevention and Control, the day aims to raise awareness about the global and escalating issue that is antimicrobial resistance (AMR). Don Whitley Scientific acknowledges this day by exploring whether antimicrobial peptides may be a solution. 

The requirement for novel antimicrobial agents is becoming ever more urgent as AMR levels increase around the world. Due to the unsatisfactory outcomes of traditional antimicrobial treatments, alternative therapies are becoming more well-recognised, including antimicrobial peptides (AMPs) [1]. AMPs are pre-existing, naturally occurring molecules that have been universally present in the gene pool of bacteria for a very long time and are the first line of defence for a range of organisms [1]. They are proving favourable as an alternative treatment as they have broad spectrum activity against both Gram-negative and positive bacteria as well as fungi; in addition, their low immunogenicity and mutagenicity makes them a positive candidate for targeting multi-drug-resistant pathogens [1]. Research has shown that the genomics of AMPs are vital for understanding their efficacy and ability to have maximum antimicrobial activity, for example, activity is dependent on the content of lipophilic groups as opposed to the amino sequence [1]. Metagenomics has therefore become a key area of investigation.

Laverty et al. determined that the lipopeptide C12-OOWW-NH2 was one of the most potent antimicrobial agents, but as it is a synthetic compound, there is an arguable preference for more naturally occurring peptides [2]. As a result, Megaw et al. had their study published this year, which aimed to identify naturally occurring peptides with the same structural properties as C12-OOWW-NH2, but also with the same potency [1]. They selected several peptides for synthesis and then screened them against a range of bacterial strains including Pseudomonas aeruginosa, Bacillus cereus and Clostridium perfringens [1]. All strains were incubated at 37°C aerobically, except for C. perfringens which was grown in a Whitley Anaerobic Workstation with an atmosphere of 80% N2, 10% CO2, and 10% H2 [1]. Antimicrobial activity was determined using zones of inhibition and minimal inhibitory concentrations using the broth microdilution method [1]. In addition, they investigated how modifications to their peptides affected antimicrobial activity and found that extending the peptide and including a C-terminal strengthened activity [1]. A promising candidate named PB2-10aa-NH2 was identified and with modifications, there was improved thermostability and solubility as well as a synergistic effect with ethanol [1]. This is encouraging as imbedding the agent into the phospholipid bilayer has a strong correlation with antimicrobial activity [1]. Ultimately, if a natural AMP alternative to the potent synthetic AMP is available then this would be preferable, so research of this kind is extremely vital.

Favourable results in AMR research, such as those gained from metagenomic studies, provide hope that we can reduce AMR and potentially one day solve the problem once and for all.

If you are interested in reading more scientific papers, please head over to our Published Papers database. Or for more information on our anaerobic workstations, please view the products below or contact sales@dwscientific.co.uk

References:

1. Megaw J, Skvortsov T, Gori G, Dabai AI, Gilmore BF, Allen CCR. A novel bioinformatic method for the identification of antimicrobial peptides in metagenomes. Journal of Applied Microbiology. 2024 Feb 21;135(3):lxae207.

2. Laverty G, McLaughlin M, Shaw C, Gorman SP, Gilmore BF. Antimicrobial Activity of Short, Synthetic Cationic Lipopeptides. Chemical Biology & Drug Design. 2010 Mar 30;75(6):563–9.

The Evolution of Hypoxic Conditions in Cell Culture

The ways that cells are cultured have evolved significantly over time due to advancements in technology. Cell culture techniques were originally very limited and depended heavily on basic conditions. As research has progressed, the variety of methods for cell culture has also increased, including hypoxic workstations, and bioreactors. These advancements offer greater precision, improving experiments and facilitating a wide range of applications.

What did scientists use before hypoxic workstations?

Up until 1972, when it was discovered by Alan Richter and his colleagues, it wasn’t known that culturing certain cell types in low oxygen increased the number of cells that could successfully grow in a plastic dish. Later in 1977, Packer and Fuehr found that 10% oxygen drastically increased the lifespan of human fibroblasts. 

The hypoxic chamber that we know today, with its sophisticated design and precise gas control, was not introduced until the 1990s. Prior to this, scientists would have had to manually adjust environmental conditions like temperature and humidity. This was achieved by using basic incubators equipped with temperature controls and water pans. The process of manually creating low oxygen levels, however, would have been even more complex, requiring a specific mixture of nitrogen and carbon dioxide to dictate the amount of oxygen. The alternative to this would be to place cells in a sealed container before flushing with nitrogen gas to displace oxygen.

What does the modern hypoxic workstation offer?

• A controlled environment: Control gases from the switch of a button

• Safety: Hypoxic cabinets provide safer working conditions for researchers

• Research flexibility: More consistent and reliable results

• Customisation: Workstations can be tailored to the individual needs of research

The Whitley Hypoxystation

The Whitley Hypoxystation is a state-of-the-art hypoxic chamber that allows for precise control over oxygen, carbon dioxide, humidity and temperature. In total, Don Whitley Scientific (DWS) offers 10 models of the Whitley Hypoxystation, each with their own set of unique advantages, all available with HEPA filtration to ensure atmospheric cleanliness. Although DWS sold their first hypoxic workstation in 2008, we have since become a staple in the field of hypoxic research, widely recognised for reliability and advanced technology.

Some applications of the Whitley Hypoxystation include:

• Cancer research

• Neurology research

• Cardiovascular research

• Plant response to hypoxia

We love seeing the endless research possibilities that the Whitley Hypoxystation provides, especially when we can help facilitate research that has the potential to treat illnesses and save lives. 

If you’re interested in our hypoxic range, please view the products below or contact us at sales@dwscientific.co.uk

O2 Measurement for the F & B Industry? Packaging and quality control devices for non-invasive O2 measurements

Do you work in packaging development, material research, quality control or in-process control in the food & beverage industry? Then rely on our well-proofed products, such as the Fibox 4 (trace) intended for packaging and quality control. Below you will find matching application examples and please feel invited and find the full range of product solutions for the Food & Beverage Industry on our webpage.

Food & Beverage: Quality Control and Packaging

Designed as portable, handheld devices the Fibox 4 and Fibox 4 trace can easily be applied for quality control in the food & beverage sector. Together with the versatile PreSens oxygen sensors which can be integrated in pipes, containers or directly held into samples the Fibox 4 and Fibox 4 trace offer complete freedom and multiple options for application to check the quality and shelf-life of your products in different stages of the production and filling process.

Unearthing the Secrets of Anaerobes

In recent years, anaerobic bacteria have shown a rise in antimicrobial resistance (AMR), leading to the emergence of resistant strains - sometimes described as “superbugs”. This development has made it much harder to treat infections effectively, posing a global challenge. We offer a wide range of products, some of which can be used to contribute to this field of research. In this article, we focus on the Whitley A35 Anaerobic Workstation. 

Characterisation of food chain C. difficile isolates in Ireland

A study conducted in Ireland by Marcos et al. titled ‘Characterization of Food Chain Clostridioides difficile Isolates in Terms of Ribotype and Antimicrobial Resistance’ utilised the Whitley A35 Anaerobic Workstation to maintain these isolates and to perform Minimum Inhibitory Concentration (MIC) testing using the E-test method. The aim of this study was to characterise C. difficile isolates from the farm, abattoir, and retail outlets in Ireland in terms of ribotype and antibiotic resistance.

C. difficile as a drug target for microbiome-sparing antibiotics

The Whitley A35 Anaerobic Workstation was used in a recent study by Dureja et al., ‘In vivo evaluation of Clostridioides difficile enoyl-ACP reductase II (FabK) inhibition by phenylimidazole unveils a promising narrow-spectrum antimicrobial strategy’. This research investigated C. difficile enoyl ACP reductase (CdFabK), a crucial enzyme in de novo fatty acid synthesis, as a drug target for microbiome-sparing antibiotics. C. difficile R20291 was used as a control strain throughout the study, and all bacterial strains were grown at 37 °C in the Whitley A35 Workstation.

Antimicrobial Susceptibility of Bifidobacterium pseudocatenulatum in Vietnam

In a study by Chung The et al. titled ‘Exploring the Genomic Diversity and Antimicrobial Susceptibility of Bifidobacterium pseudocatenulatum in a Vietnamese Population’ the Whitley A35 Anaerobic Workstation played a role in the culturing and identifying the anaerobic bacteria, Bifidobacterium. This applied to all samples that attributed to Bifidobacterium pseudocatenulatum above 0.1% of total sequenced reads (7 adults, 9 children). This paper provides a wider understanding of the anaerobic bacteria in Vietnam and its genomic diversity, metabolic potential, and antimicrobial susceptibility.

We are delighted to see the Whitley A35 Anaerobic Workstation being used to support the ongoing research into AMR, a crucial area of research and gateway to a broad spectrum of studies. For more information on our Anaerobic Workstation range please see the products below or if you're interested in reading more scientific papers, please visit our Published Papers database.

Anaerobic Bacteria: Improving Laboratory Methods

In clinical microbiology, every minute counts when it comes to maintaining the viability of anaerobic bacteria in patient specimens. Ensuring that samples are quickly placed into anaerobic conditions is crucial to minimise the stress of oxygen exposure on anaerobic bacteria and maximise recovery of all species present in the sample.

At Don Whitley Scientific, we understand the challenges associated with culturing and maintaining fastidious anaerobes. That’s why we’ve designed our Whitley Workstations to streamline your processes and protect your samples.

For those times when samples or inoculated plates need to be quickly transferred to the anaerobic workstation from the lab bench, our innovative letterbox system can make a big difference. 

The optional single plate entry port, known as the ‘letterbox,’ allows you to quickly introduce Petri dishes, specimens and other small items into the anaerobic environment without delay. Positioned conveniently on the left-hand side of our A35 and A45 Workstations, this 15 cm entry port facilitates swift and seamless sample entry.

In contrast, the traditional anaerobic jar method often means waiting until you have enough plates to set up the jar. During this waiting period, plates are exposed to air, potentially compromising recovery of anaerobic bacteria. With the letterbox option on our workstations, you can avoid this issue and maintain the quality of your samples.  There is also the possibility of transferring samples directly into the workstation for anaerobic processing and inoculation of pre-reduced culture media, in cases where stringent handling of specimens is desirable.

By integrating our Whitley Workstations into your lab, you’re choosing a solution that helps you uphold best practices in anaerobic microbiology, ensuring repeatable and accurate results every time.

If you are using anaerobic jars then consider operating them with the Whitley AtmoGen, a jar gassing solution that allows you to rapidly generate strict anaerobic conditions with a much reduced cost per jar in comparison with disposable systems. AtmoGen also increases the speed at which anaerobiosis is attained inside a jar and reduces the exposure of inoculated plates to oxygen.

Discover how our innovative solutions for anaerobic atmospheres can transform your lab work by scheduling a demo today!

High-throughput Characterization of Fungal Pathogens and Monitoring Phenotypic Drift in Bacteria

ATCC, a leading supplier of authenticated biological materials, is partnering with Biolog, Inc. to monitor phenotypic drift in bacterial strains as well as characterize WHO listed fungal pathogens. Understanding the phenotypic profile of a given microbe is key to contextualizing the results of any study. Here, we generated baseline phenotypic profiles for eleven fungal pathogens recognized by the WHO to be of special concern in the global health community and common production and research strains of bacteria. To investigate the occurrence of phenotypic drift in laboratory and production strains we then simulated years of passages while monitoring for changes in each organism’s phenotypic profile. Once phenotypic drift was captured, affected generations were then sequenced to ascertain the occurrence of mutations within each strain. This study demonstrated that high-throughput phenotypic screening is a valuable tool for characterizing strains and monitoring phenotypic instability due to genetic changes due to passaging, highlighting the importance of using authenticated strains and minimizing passaging for reliable bioproduction and experiment.

Studying microbial communities: a review of EcoPlate use cases and the newest community analysis features in Odin Software

With three decades of use and over 1,400 published papers, Biolog’s EcoPlates have become a well-established and powerful method for evaluating the functional diversity of microbial communities. EcoPlates can be used to characterize functional traits of microbial populations from soil or water, for example, under either aerobic or anaerobic conditions. This facilitates the quantification of the metabolic capabilities of microbial communities across any natural or perturbed environment. This webinar will describe recent publications where EcoPlates have been employed in top-down and bottom-up approaches to answer questions in basic and applied microbial ecology. We will also review the new Community Analysis features in Odin software, inspired by EcoPlate users. Incorporating the richness of kinetic data obtained with the Odin family of instruments, this platform provides an ideal method for understanding the metabolic profiles of microbiome samples, and how diverse populations can change over time and with various treatments.

Biolog delves into characterizing gut micobiomes with the launch of PreBioM plates

June 11, 2024 — Hayward, CA – Biolog, Inc., a leading provider of cell-based microbial identification and phenotypic and metabolic profiling products and services,

is excited to announce the launch of its new PreBioM™ microplate product line for microbiome analysis and organism selection. These microplates are designed to enable phenotypic characterization of individual organisms and communities typically found in the gut microbiome, providing deeper insights into microbial behavior and interactions, and allowing researchers to study and produce new probiotics.

The PreBioM plates feature a curated selection of prebiotic substrates, ranging from simple sugars to complex dietary fibers. The diverse array of substrates, pre-arrayed on the plates, offer a powerful tool for interrogating the influence of different prebiotics on microbial growth and activity.

“Understanding the metabolic capabilities of microbial communities is crucial for advancing our knowledge of the gut microbiome and its impact on health.” said Robert Wicke, CEO of Biolog. “PreBioM plates provide researchers with an unprecedented ability to explore metabolic pathways of bacterial communities, opening new avenues for discovery in this field.”

Leveraging the same technology as Biolog’s well-established EcoPlateTM microplate, which has been extensively used for studying various types of microbiome communities in soil, water, and wastewater, the PreBioM plates extend community-level analysis capabilities into the realm of human and animal microbiomes. With the launch of these new plates, the first of many to come, Biolog reaffirms its commitment to advancing the understanding of microbes with a suite of instruments, software, microplates, and services.

“One of the key benefits of our phenotyping plates is their versatility. They can be used under aerobic and anaerobic conditions, making them suitable for a wide range of studies that impact discovery through production of critical probiotic strains,” said Wicke. “When used in conjunction with Biolog’s Odin™ platform, PreBioM plates enable functional characterization of living communities of organisms, with data analysis tools that mine the richness of kinetic data, providing insight into how diverse populations change over time or in response to various treatments.”

About Biolog

Biolog has the tools, services, and support to provide the microbial phenotypic characterization and polyphasic identification capabilities required to produce functionally relevant insights. Our products and services enable characterization of microbial and mammalian cells as well as identification of thousands of species of bacteria, yeast, and fungi. Learn more at www.biolog.com.

Contact:
Biolog Contact:
John Proctor, Ph.D.
CCO
jproctor@biolog.com
408-306-0414

Cells, Cancer Research and Radiotherapy: Sheffield University

At Don Whitley Scientific, we are always excited to hear how our workstations contribute to ground breaking research. Recently, we had the pleasure of speaking with Diane Lefley, a PhD student at the University of Sheffield, about her innovative project using our Whitley H35 Hypoxystation. Diane's work focuses on a promising approach to treating breast cancer bone metastases, combining Radium-223 with DNA damage repair inhibitors.

The Project: A Dual Approach to Cancer Treatment

“My project is about treating breast cancer bone metastases with a combination of Radium-223 and DNA damage repair inhibitors. Radium-223 mimics calcium and rapidly binds to bone in vivo, irradiating close to the bone surface. If tumours are small when Radium-223 treatment is given, then the tumour will be completely irradiated. If the bone tumour is larger, not all the tumour will be effectively irradiated, meaning some cells will be able to repair their DNA damage. The DNA damage repair inhibitor should prevent this, sensitising the cells to radiation, i.e. a small dose of radiation is needed to kill the cells when the DNA damage repair inhibitor is used.

The in vitro side of my project will all take place in hypoxia, as the bone is a hypoxic environment.

My experiments are currently 7 days for MTT assays and for protein – this is so the cells can cycle (or have their cycle halted by inhibitors) multiple times for the effect to become noticeable. It takes around 72 hours for the DNA damage repair inhibitors to have a significant effect on cell cycle, but mitochondrial stress then increases, which interferes with the MTT assay, which is why I wait longer. I will also look at clonogenic assays which will take 10-14 days depending on the cell line.”

Why Hypoxia Matters

A key element of Diane’s research is the use of hypoxic conditions, replicating the bone environment where oxygen levels are low. She typically uses a 3% oxygen concentration for her experiments. This is because, “bone oxygen concentration is 1-6% and 3% lies in the middle and, the bone metaphysis where tumour cells home to is 2.5% oxygen. Large tumours would be further from the metaphysis.”

“Hypoxia is important for two reasons: 1.Hypoxia activates proteins involved in the DNA damage repair pathway that are also involved in halting cell cycle (cells proliferate less in hypoxia due to these proteins being activated), therefore DNA damage repair inhibitors could have more of an effect in hypoxia – the inhibitors would inactivate these proteins and force the cycle to cell with DNA damage, leading to higher chance of apoptosis and 2. In hypoxia, there are fewer free radicals available to cause DNA damage from radiation. In summary, the cells are less sensitive to radiation and more sensitive to DNA damage repair inhibitors in hypoxia.”

The Benefits of Using a Whitley Hypoxystation

When compared with a CO2 incubator the Hypoxystation allows the cells to remain within a hypoxic environment. “It is essential that the cells are hypoxic when the Radium-223 is added. The cabinet also keeps the radiation contained. The Perspex on the front of the unit is thicker than the shielding I was previously using, and in case of a spill everything is contained.”

“Oxygen can be cycled within the Hypoxystation, which may be useful as proteins in DNA repair are highly activated upon reoxygenation to prevent cell cycle with DNA damage and apoptosis. I am not sure if we will use it in this project, but we may use it in the future. I also like that you can track the oxygen concentration so you can see when it becomes stable and check the concentration while you have been away from the lab.”

Why choose Don Whiley Scientific?

“I met Stuart Moore, a DWS sales representative at the Nottingham BACR and told him of my project, but I was on a budget as I was doing a PhD. Stuart worked with me to ensure I would have a cabinet to facilitate my research that gave me the features I needed within budget. I personally think your customer service is better than your competitors!”

While I haven’t had any service or maintenance yet, the overall team that installed the Hypoxystation did a great job.”

We thank Diane for providing such a detailed account of her work and wish her all the best with her continued research. If you have an application that requires hypoxic conditions, please email us at sales@dwscientific.co.uk or have a look at the products below.

What is Osteomicrobiology?

In recent years, the term "Osteomicrobiology" has surfaced to describe the rapidly emerging research field looking into the role of the gut microbiome on bone metabolism and health. This research field aims to bridge the gaps between bone physiology, gastroenterology, immunology, and microbiology (1).

Okoro et al (2023) recently evaluated the link between the gut microbiome and markers of bone health (bone density, the microarchitecture and strength) by conducting further work on previous studies involving 2 cohorts of participants [2]. The Framingham heart (FHS) study participants were aged between 32 and 89 years and were recruited from a town in Massachusetts. The osteoporotic fractures in men (MrOS) study recruited men at six clinical sites in the U.S between 2000-2002 who were 78 to 98 years of age [2]. Stool samples were donated by participants in both studies and were analysed by sequencing of the 16s rRNA, grouped by taxa and converted into relative abundance. For both cohorts, 8 out of the top 10 most abundant genera were identical, however Akkermansia and Escherichia/Shigella were more abundant in the MrOS group and Agathobacter and Roseburia were more abundant in the FHS group [2]. The total variability between the microbiomes of the 2 cohorts was found to be 3%.  Furthermore, bone health was monitored by high resolution peripheral quantitative computed tomography (HR-pQCT) at the distal radius and tibia of the participants, including measurements of volumetric bone mineral density (vBMD) and cortical bone area. The demographics and clinical characteristics were compared and corrected for statistically between the 2 studies including sex, ethnicity, medications, and age. Participants were excluded who had undergone colon surgery or had taken antibiotics in the previous 12 months.

In the FHS cohort, 29 advantageous and 38 non advantageous microbiome-bone associations were found involving 37 genera and 18 HR-pQCT measurements. They identified that Turicibacter and Victivallis abundance had greater radius and tibial bone area and vBMD. Blautia, Akkermansia and Marvinbryantia abundance was negatively associated with cortical thickness, cortical bone area to total area ratio and vBMD [2]. In the MrOS cohort, they found five microbiome-bone associations [2]. Increased abundance of Methanobrevibacter was associated with lower cortical vBMD, while increased abundance of Lachnospiraceae NK4A136 group was associated with greater cortical bone mass and vBMD [2]. Higher abundance of Cloacibacillus was positively associated with greater vBMD [2]. Across both cohorts, the group identified a strain called DTU089 and found consistency in bone measurements that in higher abundance, the strain was associated with lower tibia cortical bone mass and vBMD [2]. 

Future studies are needed to determine if the gut microbiome composition is a novel therapeutic target for biomarkers of bone health, monitoring of conditions such as osteoporosis and predictions of fractures [1] and potential probiotics.

Are you developing a live biotherapeutic product? Our range of anaerobic workstations provide excellent conditions for the processing, incubation and examination of samples without exposure to atmospheric oxygen. Check out the Workstation Builder tool to configure your own custom suite of isolators!

Written by DWS microbiologist Charlotte Austin

References

1. Ohlsson, C., & Sjögren, K. (2018). Osteomicrobiology: A New Cross-Disciplinary Research Field. Calcified tissue international, 102(4), 426–432.

2. Okoro P, Orwoll E,Huttonhower C, Morgan X, Kuntz T,  A two-cohort study on the association between the gut microbiota and bone density, microarchitecture, and strength

Submerge Completely ... with our new versions of the OXYBase-SW

Can cancer researchers benefit from mimicking the body in their labs?

The tumour microenvironment, characterized by low oxygen levels (hypoxia), has been shown to impact cellular processes, including machinery functionality, viability, and gene/protein expression. Understanding the influence of hypoxia, often termed 'tumour physioxia,' is crucial in the research, discovery, and development of oncology treatments. Hypoxia's association with drug resistance and therapeutic inefficacy underscores its importance as a pivotal factor to address in oncological studies and treatment strategies.

Solid tumours, notorious for their adaptability, thrive within hypoxic microenvironments, fostering cellular resilience and treatment resistance. This necessitates a deeper understanding of cellular responses to oxygen deprivation for the advancement of oncological interventions. We uncover the profound implications of replicating in vivo physioxic conditions within the laboratory setting.

Study 1: Hypoxia's Influence on Tumour Biology

van de Schootbrugge et al. examined the impact of hypoxia on αB-crystallin expression in head and neck squamous cell carcinoma (HNSCC), uncovering its role in promoting cell survival under physioxia. αB-crystallin was more abundantly present in hypoxic areas of HNSCC biopsies than in normoxic areas whilst decreased αB-crystallin levels reduced cell survival under hypoxic conditions. Their findings shed light on the importance of mimicking the tumour microenvironment to reveal key survival mechanisms of cancer cells.

Study 2: Hypoxia and Treatment Resistance

Alam et al. investigated the contribution of HIF2α to antiestrogen resistance in breast cancer cells. Determining if there was any connection between hypoxia and therapeutic outcomes. Their findings highlighted hypoxia diminishes the effectiveness of antiestrogen treatments (tamoxifen and fulvestrant) in ERα-positive breast cancer cells, coinciding with elevated HIF2α expression and increased resistance to therapy Studying cells under physiologically relevant oxygen conditions to unravel treatment resistance mechanisms is a vital consideration for research design, drug development and accurate pharmacodynamic studies.

Study 3: Modulating Tumour Aggressiveness

Campbell et al. explored the impact of ascorbate availability on tumour behaviour under hypoxic conditions, revealing its role in modulating HIF1 activity and tumour aggression. Their study demonstrated a negative correlation between ascorbate availability and HIF1 response in LL/2 cells, indicating a potential role of ascorbate in modulating tumour aggression.  The findings of this study present an argument for past, present and future therapies to be assessed under hypoxic conditions to improve the understanding and potential efficacy of cancer therapies for improved patient outcomes.

Study 4: Considerations for Accurate Research Under Hypoxia

Carreau et al. presented the critical role of oxygen partial pressure (pO2) in human tissues and its significance in maintaining physiological balance. Focusing closely on the impact of hypoxia on various organs and tissues. Particularly in pathological conditions such as cancer. The investigators emphasised 5 key considerations for performing accurate research under hypoxia:

• Physioxia Importance: Studying cells under physiologically relevant oxygen levels is crucial for accurately reflecting cellular behaviours and responses in various tissues and organs.

• Tumour Biology: Understanding the role of hypoxia in tumour microenvironments is essential for evaluating therapy efficacy and developing targeted treatment strategies.

• Experimental Relevance: By mimicking the oxygen levels found in specific organs, researchers can better understand the impact of oxygen on gene expression, proteome regulation, and overall cell activity.

• Experimental Accuracy: Understanding the impact of oxygen levels on cellular activities is essential for interpreting experimental results accurately and translating findings from in vitro studies to real-life clinical settings.

• Clinical Translation: Utilizing physiologically relevant oxygen conditions in experimental settings can enhance the translation of in vitro findings to real-life clinical scenarios, improving the relevance and applicability of research outcomes.

As researchers navigate the complexities of cancer, it becomes increasingly clear that mimicking the tumour microenvironment in the lab is not only advantageous but essential. By embracing physioxic conditions, new insights into tumour biology, treatment resistance mechanisms, and therapeutic strategies will be discovered.

References

1. Alam, M., Persson, C., Reinbothe, S., Kazi, J., Rönnstrand, L., Wigerup, C., . . . Jögi, A. (2016). HIF2α contributes to antiestrogen resistance via positive bilateral crosstalk with EGFR in breast cancer cells. Oncotarget, 7(10), 11238-11250. doi:10.18632/oncotarget.7167

2. Campbell, E., Vissers, M., & Dachs, G. (n.d.). Ascorbate availability affects tumor implantation-take rate and increases tumor rejection in Gulo-/- mice. Hypoxia (Auckl)., 8(4), 41-52. doi:10.2147/HP.S103088

3. Carreau, A., El Hafny-Rahbi, B., Matejuk, A., Grillon, C., & Kieda, C. (2011). Why is the partial oxygen pressure of human tissues a crucial parameter? Small molecules and hypoxia. Journal of Cellular and Molecular Medicine, 15(6), 1239-1253. doi:10.1111/j.1582-4934.2011.01258.x

4. van de Schootbrugge, C., Schults, E., Bussink, J., Span, P., Grénman, R., Pruijn, G., . . . Boelens, W. (2014). Effect of hypoxia on the expression of αB-crystallin in head and neck squamous cell carcinoma. BMC Cancer, 14(252). doi:10.1186/1471-2407-14-252

Can cancer researchers benefit from mimicking the body in their labs?

The tumour microenvironment, characterized by low oxygen levels (hypoxia), has been shown to impact cellular processes, including machinery functionality, viability, and gene/protein expression. Understanding the influence of hypoxia, often termed 'tumour physioxia,' is crucial in the research, discovery, and development of oncology treatments. Hypoxia's association with drug resistance and therapeutic inefficacy underscores its importance as a pivotal factor to address in oncological studies and treatment strategies.

Solid tumours, notorious for their adaptability, thrive within hypoxic microenvironments, fostering cellular resilience and treatment resistance. This necessitates a deeper understanding of cellular responses to oxygen deprivation for the advancement of oncological interventions. We uncover the profound implications of replicating in vivo physioxic conditions within the laboratory setting.

Study 1: Hypoxia's Influence on Tumour Biology

van de Schootbrugge et al. examined the impact of hypoxia on αB-crystallin expression in head and neck squamous cell carcinoma (HNSCC), uncovering its role in promoting cell survival under physioxia. αB-crystallin was more abundantly present in hypoxic areas of HNSCC biopsies than in normoxic areas whilst decreased αB-crystallin levels reduced cell survival under hypoxic conditions. Their findings shed light on the importance of mimicking the tumour microenvironment to reveal key survival mechanisms of cancer cells.

Study 2: Hypoxia and Treatment Resistance

Alam et al. investigated the contribution of HIF2α to antiestrogen resistance in breast cancer cells. Determining if there was any connection between hypoxia and therapeutic outcomes. Their findings highlighted hypoxia diminishes the effectiveness of antiestrogen treatments (tamoxifen and fulvestrant) in ERα-positive breast cancer cells, coinciding with elevated HIF2α expression and increased resistance to therapy Studying cells under physiologically relevant oxygen conditions to unravel treatment resistance mechanisms is a vital consideration for research design, drug development and accurate pharmacodynamic studies.

Study 3: Modulating Tumour Aggressiveness

Campbell et al. explored the impact of ascorbate availability on tumour behaviour under hypoxic conditions, revealing its role in modulating HIF1 activity and tumour aggression. Their study demonstrated a negative correlation between ascorbate availability and HIF1 response in LL/2 cells, indicating a potential role of ascorbate in modulating tumour aggression.  The findings of this study present an argument for past, present and future therapies to be assessed under hypoxic conditions to improve the understanding and potential efficacy of cancer therapies for improved patient outcomes.

Study 4: Considerations for Accurate Research Under Hypoxia

Carreau et al. presented the critical role of oxygen partial pressure (pO2) in human tissues and its significance in maintaining physiological balance. Focusing closely on the impact of hypoxia on various organs and tissues. Particularly in pathological conditions such as cancer. The investigators emphasised 5 key considerations for performing accurate research under hypoxia:

• Physioxia Importance: Studying cells under physiologically relevant oxygen levels is crucial for accurately reflecting cellular behaviours and responses in various tissues and organs.

• Tumour Biology: Understanding the role of hypoxia in tumour microenvironments is essential for evaluating therapy efficacy and developing targeted treatment strategies.

• Experimental Relevance: By mimicking the oxygen levels found in specific organs, researchers can better understand the impact of oxygen on gene expression, proteome regulation, and overall cell activity.

• Experimental Accuracy: Understanding the impact of oxygen levels on cellular activities is essential for interpreting experimental results accurately and translating findings from in vitro studies to real-life clinical settings.

• Clinical Translation: Utilizing physiologically relevant oxygen conditions in experimental settings can enhance the translation of in vitro findings to real-life clinical scenarios, improving the relevance and applicability of research outcomes.

As researchers navigate the complexities of cancer, it becomes increasingly clear that mimicking the tumour microenvironment in the lab is not only advantageous but essential. By embracing physioxic conditions, new insights into tumour biology, treatment resistance mechanisms, and therapeutic strategies will be discovered.

References

1. Alam, M., Persson, C., Reinbothe, S., Kazi, J., Rönnstrand, L., Wigerup, C., . . . Jögi, A. (2016). HIF2α contributes to antiestrogen resistance via positive bilateral crosstalk with EGFR in breast cancer cells. Oncotarget, 7(10), 11238-11250. doi:10.18632/oncotarget.7167

2. Campbell, E., Vissers, M., & Dachs, G. (n.d.). Ascorbate availability affects tumor implantation-take rate and increases tumor rejection in Gulo-/- mice. Hypoxia (Auckl)., 8(4), 41-52. doi:10.2147/HP.S103088

3. Carreau, A., El Hafny-Rahbi, B., Matejuk, A., Grillon, C., & Kieda, C. (2011). Why is the partial oxygen pressure of human tissues a crucial parameter? Small molecules and hypoxia. Journal of Cellular and Molecular Medicine, 15(6), 1239-1253. doi:10.1111/j.1582-4934.2011.01258.x

4. van de Schootbrugge, C., Schults, E., Bussink, J., Span, P., Grénman, R., Pruijn, G., . . . Boelens, W. (2014). Effect of hypoxia on the expression of αB-crystallin in head and neck squamous cell carcinoma. BMC Cancer, 14(252). doi:10.1186/1471-2407-14-252

With An Eye on the Future, Biolog Launches Odin

June 15, 2023 — Hayward, CA – Biolog, Inc., a leading provider of microbial polyphasic analysis, today announced the launch of the Odin™ system, an all-in-one cellular phenotyping and microbial identification platform. Odin is compatible with all Biolog assays and is available now.

Until today, Biolog’s OmniLog® platform was the gold standard for researchers who want actionable insights on microbial metabolic phenotypes. Odin now builds on that foundation while maintaining the core capabilities to incubate, analyze, and compare thousands of microbial and mammalian growth conditions in a single experiment, and to identify aerobic bacteria by its metabolic fingerprint.

Odin changes up the detection method and uses Optical Density (OD), which enables a critical additional modality to measure cell growth. Now on a single platform, researchers can distinguish the effect of a substrate on growth (by OD) and by respiration using our proprietary dye, now an absorbance-based measurement. With the additional Odin capabilities, researchers can now decouple a microbe’s respiration rate from its division rate and more fully understand the metabolic output of a cell. With a fully loaded system, Odin records the OD for all conditions for up to 50 plates at a time, measuring the kinetics of cell growth and respiration for thousands of conditions in parallel.

Odin also has broader identification capabilities, including anaerobes, yeast, and filamentous fungi. All told, Odin can identify thousands of microbes, allowing researchers to classify unknown organisms and phenotype them on the same platform.

“We heard from our customers the need to directly measure growth. In a short time, we brought in additional scientific expertise, and built a product development engine. The customer-focused mindset and our dedicated teams have enabled us to quickly meet the needs of our customers,” said Robert Wicke, CEO of Biolog.

“Understanding how microbes respond to their environment and food sources is the major economic driver in sustainable agriculture, bioprocessing and synthetic biology, to name just a few areas. We are enthusiastic about the foundation that Odin creates for Biolog and our customers.” said Mr. Wicke.

About Biolog

Biolog has the tools, services, and support to provide the microbial phenotypic characterization and polyphasic identification capabilities required to produce functionally relevant insights. Our products and services enable characterization of microbial and mammalian cells as well as identification of thousands of species of bacteria, yeast, and fungi. Learn more at www.biolog.com.

Contact:
Biolog Contact:
John Proctor, Ph.D.
CCO
jproctor@biolog.com
408-306-0414

Biolog expands the Odin family, and access to cellular characterization, with the launch of Odin V111

April 16, 2023 — Hayward, CA – Biolog, Inc., a leading provider of cell-based microbial identification and phenotypic and metabolic profiling products and services, today announced the launch of Odin™ VIII, the newest member of its Odin family of instruments. Designed to meet the needs of laboratories with mid- throughput requirements, Odin VIII is a smaller, yet equally powerful version of the Odin™ system introduced last year.

Biolog is thrilled to offer Odin VIII as a versatile solution for laboratories seeking high-performance cellular analysis capabilities in a more compact form. “Following the success of Odin in the market, we recognize the need to cater to laboratories with varying throughput requirements,” said Robert Wicke, CEO of Biolog. “With Odin VIII, we are excited to extend the power and efficiency of the Odin platform to a broader range of users, enabling them to characterize cells and identify unknown organisms with unmatched ease and flexibility.”

Odin VIII retains the all-in-one functionality that has made Odin a preferred choice among researchers studying how microbes respond to their environment. Leveraging the same detection methods and the ability to read and analyze any Biolog microplate, Odin VIII enables the measurement of both respiration and growth dynamics, providing valuable insights into cellular physiology and metabolic activity. Odin VIII can simultaneously incubate and read up to 8 plates at once, offering the ability to streamline workflows and enhance productivity without compromising on performance. The original Odin instrument is being renamed to Odin L, to reflect its 50-plate capacity and unmatched scalability, especially important for labs with multiple users or those screening many strains at the same time. Both Odin L and Odin VIII are powered by the same Odin analytical software and share the same identification databases.

With the launch of Odin VIII, Biolog reaffirms its commitment to customer-centric solutions, empowering laboratories of all sizes to accelerate their microbial identification and characterization efforts with confidence. “This smaller system is well suited for academic research labs, fermentation departments that want to characterize or QC strains, or for researchers in sustainable agriculture studying microbiome communities. Additionally, for our customers whose environmental monitoring needs have grown, but still don’t quite need the full capacity of an Odin L, we now have a medium throughput solution for in-house identification of bacteria, anaerobes, yeast, and filamentous fungi. We remain committed to empowering scientists with the tools they need to unlock the mysteries of microbial behavior,” said Wicke.

About Biolog

Biolog has the tools, services, and support to provide the microbial phenotypic characterization and polyphasic identification capabilities required to produce functionally relevant insights. Our products and services enable characterization of microbial and mammalian cells as well as identification of thousands of species of bacteria, yeast, and fungi. Learn more at www.biolog.com.

Contact:
Biolog Contact:
John Proctor, Ph.D.
CCO
jproctor@biolog.com
408-306-0414

Biolog Announces New Leadership Team and Secures Major Investment to Drive Widespread Adoption of Industry-Leading Microbial Identification and Phenotypic Profiling Products

May 12, 2022 — Hayward, CA – Biolog, Inc., a leading provider of cell-based microbial identification and phenotypic profiling technologies, today announced the appointment of a new leadership team including industry veteran Robert Wicke as Chief Executive Officer. Former CEO and Founder Barry Bochner, Ph.D., will remain with the company part-time as its Chief Scientific Advisor. The re-incorporated company also secured a major investment from BroadOak Capital Partners and Research Corporation Technologies (RCT) and will use the proceeds to advance its cellular analysis product portfolio.  With more than 1,700 customers worldwide, the company remains committed to developing and selling its products in diverse markets including pharmaceutical QC/R&D, academic and basic research, biotechnology, environmental monitoring, agriculture, and plant and animal health.

Joining Mr. Wicke on the leadership team are John Proctor, Ph.D., as Chief Commercial Officer, Ariel Notcovich as Chief Technical Officer & Chief Operations Officer, and Jason Novi as Chief Financial Officer & Senior Vice President of Corporate Development.  The seasoned team has managed several other life science companies and developed world-class products that have led to great outcomes for both investors and customers.  Mr. Wicke states, “Barry built a world-class company that is well known for its microbial identification, phenotypic analysis, and metabolic profiling products for cells ranging from bacteria, yeast, and fungi, to mammalian cells and mitochondria. I am excited for this opportunity to work with John, Ariel, Jason, and the full Biolog team to execute a long-term growth strategy that will enable us to realize the full potential of our novel cell-based testing systems through new product development and commercial leadership.”

“Over the past 35 years, I have had the pleasure of working with a very experienced and dedicated scientific staff at Biolog.  Together, we were able to develop novel technologies and products that can completely change how researchers can test the properties of a cell,” said Dr. Bochner. “As the demand for our technology has continued to expand, we needed to bring in an experienced management team that could take our business to the next level. With Robert, John, Ariel, and Jason, we have a full team in place that has successfully commercialized numerous life science technologies in the past. I am excited to support them as we turn the page and write a new chapter of what is clearly going to be an exciting future for Biolog.”

Biolog will be backed and funded by growth capital specialists in the life science space, BroadOak Capital Partners and RCT and guided by a board of life sciences industry leaders.  The Board of Directors will be Robert Wicke, Bill Snider (BroadOak), Chad Souvignier (RCT), David Weber (Independent), Tim Harkness (Independent), and Joe Keegan (Independent).

About Biolog

Biolog is a leader in microbial identification, phenotypic cell profiling, and metabolic fingerprinting for microbial and mammalian cells. Located in Hayward, CA, the company’s products enable the identification of thousands of species of bacteria, yeast, and fungi as well as characterization of human cells and mitochondria. Biolog products are available worldwide, either directly from the company or through its extensive network of international distributors.  For more information, visit www.biolog.com.

Biolog Contact:
John Proctor, Ph.D.
CCO
jproctor@biolog.com
408-306-0414

First Whitley Laminar Flow Workstation Launched

Don Whitley Scientific is thrilled to announce the launch of the new Whitley A135 Laminar Flow Workstation. For over 40 years, DWS has enjoyed a reputation as one of the world’s leading manufacturers of anaerobic workstations. As the needs of our customers have evolved over the last four decades - so too have our products.

Our Anaerobic Range comes in all shapes and sizes; the most compact can accommodate 200 Petri dishes, whilst the largest can house up to 1,400. For work which requires the ability to operate large pieces of laboratory equipment in a strictly controlled anaerobic environment, the A135 and A155 offer internal volumes of 950 and 1,900 litres respectively. 

The next step for DWS was to develop innovative laminar flow technology. The Whitley A135 Laminar Flow Workstation is a truly unique* product that provides highly effective product protection. Combining laminar flow technology with our expertise in designing and manufacturing anaerobic workstations, we have successfully created a laminar flow workstation that allows users to process samples under strict anaerobic conditions.

Learn more about this revolutionary product here.

HEPA Filtration Now Available on Whitley Workstations with 30 Litre Airlocks →

Don Whitley Scientific announces that all Whitley 85 and 95 sized workstations are now available with the Whitley Internal HEPA Filtration System.

The Whitley Workstation range includes models suitable for Anaerobic, Microaerobic and Hypoxic applications. Within each of these categories, there are 2 models equipped with 30 litre airlocks. The A/M/H85 Workstations can accommodate up to 610 x 90mm Petri dishes, whereas the A/M/H95 Workstations have capacity of double this amount.

With a 30 litre airlock, 90 x 90mm Petri dishes can be introduced to the workstation at once, with the airlock cycle taking only 5 minutes. In addition, the portholes on both 85 and 95 Workstations function as mini-airlocks; allowing for the introduction of 10 plates at a time.

Recently we have been working to develop variations of these models fitted with the unique Whitley Internal HEPA Filtration System, and are now pleased to announce the addition of these 6 additional variants to our range.      

With HEPA filtration, the entire atmosphere inside the workstation passes through the filter hundreds of times an hour; ensuring the chamber environment is cleaned quickly. DWS filters are located inside the workstation, which prevents the filter becoming saturated with moisture and rendered ineffective.

The internal atmosphere is circulated continuously - contributing to even temperature distribution and ensuring any internal particulates will be in motion before capture by the filter. Levels of atmospheric cleanliness inside the workstation exceed the requirements of ISO 14644 Class 3. Proof of atmospheric cleanliness can be provided on request.

WASP Touch and ProtoCOL3 play key part in Australian Lab →

A research and development lab at the New South Wales Department of Primary Industries (Sydney, Australia) recently purchased a WASP Touch Spiral Plater and ProtoCOL3 colony counter. The lab is carrying out research on food safety of fresh horticultural produce and nuts. The head of the research program, Dr S P Singh, spoke to Don Whitley Scientific to explain how these two new products, provided by DWS, are key to his team’s working processes.

Dr Singh and his colleagues are exploring fresh options when it comes to food safety: “We are developing new ways of sanitising the produce by killing potential foodborne bacterial pathogens”. The produce his lab are sanitising is often used for export, meaning this is a vital process to ensure safety for the consumer. Dr Singh explains that “we have to deal with hundreds of samples everyday” meaning “plating and enumeration is a core lab activity”.

The lab’s project was recently featured on NBN News http://www.nbnnews.com.au/2017/11/10/world-first-food-sanitiser-at-ourimbah/. The project is a world’s first and to sanitise food without the use of chemicals could be a revolutionary step forward in food safety.

Examining the Role of Autophagy in Hypoxic Tumours

Hypoxystation users Tan et al. at the University of Toronto published a paper in June examining the significance of autophagy in cancer development (“Role of Autophagy as a Survival Mechanism for Hypoxic Cells in Tumors“, Neoplasia (2016) 18, 347-355). Autophagy as a means of recycling cell components is induced under stress conditions such as hypoxia, and Tan et al. investigated the correlation of hypoxia and autophagy in solid tumours in the context of resistance to cancer therapeutics.

Cells were cultured in the H35 Hypoxystation for up to 48 hours at hypoxia (0.2 %) and compared to cells grown at ambient oxygen level. Gene silencing of autophagy proteins ATG7 and BECLIN1 with shRNA resulted in decreased cell survival under hypoxia, and inhibition of autophagy with pantoprazole exacerbated the loss of viability in the knock-down cells under hypoxia, demonstrating the cyto-protective effects of these autophagy proteins. Using the Seahorse XFe Analyzer to assess oxygen consumption in wild-type and silenced cells, Dr. Tan’s lab found reduced respiration when autophagy is disrupted, possibly due to accumulation of dysfunctional mitochondria in these mutant cells. The H35 Hypoxystation  Dr. Tan’s lab used for these studies creates a closed environment with controlled temperature, humidity, CO2 and oxygen, in which cells are cultured and manipulated over the course of days and weeks without the need to transfer into ambient conditions. The combination of an Hypoxystation and an i2 Instrument Workstation is designed to accommodate the specific requirements of the Seahorse XFe Analyzer for the duration of the metabolism assays investigating oxygen consumption and extracellular acidification.

http://www.dwscientific.co.uk/blog/examining-role-autophagy-hypoxic-tumours/