Saturday, 17 February 2018

Novel framework to infer microbial interactions


Inferring the underlying ecological networks of microbial communities is important to understanding their structure and responses to external stimuli. But it can be very challenging to make accurate network inferences. Scientists now detail a method to make the network inference easier by utilizing steady-state data without altering microbial communities.

"Existing methods require assuming a particular population dynamics model, which is not known beforehand," said Yang-Yu Liu, PhD, of the Channing Division of Network Medicine. "Moreover, those methods require fitting temporal abundance data, which are often not informative enough for reliable inference."

To obtain more informative temporal data, researchers have to introduce large perturbations to alter the microbial communities, which are not only difficult in practice but also potentially ethically questionable, especially for human-associated microbial communities. The new method developed by BWH investigators avoids this dilemma.

"The basic idea is very simple. If one steady-state sample differs from another only by addition of one species X, and adding X brings down the absolute abundance of Y, then we can conclude X inhibits the growth of Y," said Liu. The team showed that this simple idea can be extended to more complicated cases where steady-state samples differ from each other by more than one species. They verified that, if enough independent steady state data were collected from the microbial communities, then the microbial interaction types (positive, negative and neutral interactions) and the structure of the network could be inferred without requiring any population dynamics modeling. 

The method proposed by the team resembles other network reconstruction methods based on steady-state data, but unlike the previous methods, no perturbations are required to be applied to the system. Furthermore, a rigorous criterion was established by the team to check if any given steady-state data was consistent with the Generalized Lotka-Volterra (GLV) model -- a classical population dynamics model in ecology that mathematically describes the relationships between species. The team found that if the microbial community followed the GLV model, then the steady-state data could also be used to deduce the model parameters -- interspecies interaction strengths and growth rates.

The method was systematically validated using simulated data generated from different classic population dynamics models with various levels of complexity. Then it was applied to real data collected from four different synthetic microbial communities, finding that the inferred ecological networks either agree well with the ground truth or can predict the response of systems to perturbations.


Additional insights into microbial ecosystems will emerge from a better understanding of their underlying ecological networks. Inferring ecological networks of human-associated microbial communities using the method developed here will facilitate the design of personalized microbe-based "cocktails," as the authors write, to treat diseases related to microbial dysbiosis.

"I am quite excited about this method, because it may pave the way to mapping more complex microbial communities such as the human gut microbiota, which in turn will help us design better microbiome-based therapies," said Liu.

See:

Yandong Xiao, Marco Tulio Angulo, Jonathan Friedman, Matthew K. Waldor, Scott T. Weiss, Yang-Yu Liu. Mapping the ecological networks of microbial communitiesNature Communications, 2017; 8 (1) DOI: 10.1038/s41467-017-02090-2

Posted by Dr. Tim Sandle

Friday, 16 February 2018

Good Distribution Practice deficiency data


The UK MHRA has published its latest review on Good Distribution Practice (GDP) deficiency data.

The GDP Inspectorate has reviewed the GDP inspection deficiency data for 2016 to allow identification of:

·         Severity and frequency of deficiencies associated with EU GDP references;
·         High impact and high frequency issues.

The purpose of publishing the inspection deficiency data is to allow industry to perform its own assessment against the deficiency findings as part of self-inspection and continuous improvement activity.

See:MHRA



Posted by Dr. Tim Sandle

Thursday, 15 February 2018

Monoclonal antibody therapies for veterinary use


The European Medicines Agency’s (EMA) Committee for Medicinal Products for Veterinary Use (CVMP) has approved the first ever guidance at European Union (EU) level for monoclonal antibody therapies for veterinary use. The guidance was prepared by the CVMP’s Ad Hoc Expert Group on Veterinary Novel Therapies (ADVENT) in the form of a question-and-answer document.

The guidance relates to particularities of monoclonal antibodies for veterinary use, quality control for potential contaminants, stability testing, reproductive safety studies and data to address potential for indirect adverse effects.

Monoclonal antibodies are immune proteins that recognise and bind to a specific target protein, and have not been used in veterinary medicines until recently. In human medicine, these therapies have been authorised for many years for use against cancer and diseases affecting the immune system, such as rheumatoid arthritis. Therapies that are new to veterinary medicine face particular challenges due to a lack of regulatory guidance.

To access the guidance, see EMA.

Posted by Dr. Tim Sandle

Wednesday, 14 February 2018

Annual report of the Good Manufacturing and Distribution Practice Inspectors Working Group


A new document from the European Medicines Agency.

The document is the annual report of the GMP/GDP Inspectors Working Group (GMDP IWG) for the year 2016. This group was established at EMA in 1996.

The GMDP IWG provides input and recommendations on all matters relating directly or indirectly to Good Manufacturing Practice (GMP) and Good Distribution Practice (GDP).
The GMDP IWG focuses on harmonisation and co-ordination of GMP and GDP related activities at EU level. The group's role and activities are described in more detail in its mandate, which was revised in 2013.

This annual report is set out in line with the format and objectives of the 2016 work plan.

To access the document, see EMA.



Posted by Dr. Tim Sandle

Tuesday, 13 February 2018

Ocean Transport is Gaining Momentum in Pharmaceutical Logistics


One of the biggest challenges in the pharmaceutical industry has nothing to do with the drugs themselves — it’s in getting the drugs, medications and equipment from their factories to hospitals and pharmacies around the world in a timely and cost-effective manner. Trucks work for domestic transportation but run into trouble when it comes to international transport. Planes can easily cross oceans, but the cost of loading up a cargo plane with pharmaceuticals is astronomical — a cost that is often passed along to the consumer.

A special report by Megan Ray Nichols
For these reasons, ocean transportation is growing in popularity among pharmaceutical companies around the globe. What are the pros and cons of ocean transport, and is it something that you should consider for your own logistical needs?

The Speed of Ocean Transit

One of the things that has discouraged pharmaceutical companies from utilizing ocean transport in the past is the fact that it takes a while for a cargo ship to traverse the oceans between destinations. Items that required refrigeration often couldn’t be transported by sea because of a lack of refrigeration technology in the enormous cargo containers.

While speed hasn’t changed in recent years, storage technology has advanced dramatically. Specifically, it has become easier to obtain the equipment needed to create refrigerated cargo containers, allowing even temperature-sensitive pharmaceuticals to be shipped by sea without impacting the efficacy of the products.

Changes in shipping technology have enabled more and more companies to shift their shipping logistics from air to sea transport while maintaining their timelines.

Changes in Packaging and Shipping

Potential exposure to sea water and salt air is necessitating one major change — a shift from traditional wooden pallets to plastic ones.

Wooden pallets are probably the most common type of pallet used for shipping pharmaceuticals, foodstuffs and the majority of other products packed and shipped around the world. They’re popular because they’re cheap and sturdy, and they can be recycled when they start to break down. Add sea air and salt water to the mix, and these wooden pallets can become a liability, potentially breaking down faster. In addition, the humid air can encourage mold or mildew growth on the pallets themselves, creating a potential for contamination.

In the United States alone, more than two billion pallets are used every day to transport goods — and between 90 and 95 percent of them are made of wood. It stands to reason that the majority of pallets being used to ship pharmaceuticals are likely wood. If sea transit becomes the norm for pharmaceutical shipment, the use of wooden pallets could become a problem. Transitioning to molded plastic pallets, which are resistant to both weather and mold/mildew growth could help prevent this issue before it has a chance to manifest.

The Problem of Security


The pharmaceutical industry is a massive one — it’s estimated that pharmaceuticals alone make up more than $1.1 trillion in the world economy, with the number expected to grow by 2020 by 41 percent. It makes sense that this multi-billion dollar industry could potentially become the target for thieves and pirates.

Cargo theft is not a new concept — as long as there has been cargo to be transported, there have been people trying to steal it. In the past, it was on horseback and from the decks of pirate ships. Today, it’s in cars…and on the decks of more advanced pirate ships.

Keeping cargo manifests and shipping schedules secure for sea transport can help to reduce the chance of theft on the high seas, but it’s not a perfect solution. High tech solutions, such as biometric locks on shipping containers, can help make your shipments less desirable. While they can be circumvented, it takes a lot of extra steps, and it’s often not worth the effort to the potential thieves.

Tracking your shipments with technology can also help you thwart potential crime, either by discouraging thieves from committing their crime in the first place or allowing you to recover stolen goods.

Taking to the high seas isn’t just for pirates anymore. It is a fantastic way to reduce costs and keep up with your deadlines while ensuring that your products are delivered to the places that need them most.

Yeast engineered to create protein pharmaceuticals


The market for pharmaceuticals that mimic the body's own proteins -- protein-based therapeutics -- is exploding. Some of them are relatively simple to manufacture in yeast-based cell factories. Insulin and HPV vaccine are two examples that are already under production, but other therapies, such as antibodies to various forms of cancer, are significantly more difficult to manufacture.

In collaboration with Associate Professor Dina Petrovic and Mathias Uhlén's research team at the Royal Institute of Technology in Stockholm, Jens Nielsen has been mapping out the complex metabolism of yeast cells for four years.

"We've been studying the metabolism of a yeast that we already know is a good protein producer. And we found the mechanisms that can be used to make the process even more efficient. The next step is to prove that we can actually produce antibodies in such quantities that costs are reduced."

The discussion has mainly been about cancer, but there are many other diseases, for example Alzheimer's, diabetes and MS, that could potentially be treated by yeast-based protein therapies. How distant a future are we talking about?

"Our part of the process is fast, but pharmaceuticals always take a long time to develop. It could be a possibility in five years, but should absolutely be on the market in ten," Nielsen says.

He adds: “"Yeast is a superb modelling system. Almost everything in yeast is also found in humans. We have complete computer models of the metabolism of yeast, and we use the same type of models to study human metabolism.”

See:

Mingtao Huang, Jichen Bao, Björn M. Hallström, Dina Petranovic, Jens Nielsen. Efficient protein production by yeast requires global tuning of metabolismNature Communications, 2017; 8 (1) DOI: 10.1038/s41467-017-00999-2

Posted by Dr. Tim Sandle

Monday, 12 February 2018

Validation Requirements for Gaseous Sterilization Using Ethylene Oxide



Gaseous sterilization is long-established but it is increasingly being reassessed and adopted due to the growth in the markets for medical devices and for single-use sterile disposable technologies for pharmaceuticals. Gaseous sterilization is not an exact term and there are different types of gaseous sterilization (and a further distinction between gases and vapors). The first division is with the agents and here sterilizing gases include: formaldehyde, ethylene oxide, propylene oxide, ozone, and chlorine dioxide. A second division is with vapors, such as peracetic acid and hydrogen peroxide. In drawing out the gas-vapor distinction, a gas is a substance that has a single defined thermodynamic state at room temperature; in contrast, a vapor is a substance that is a mixture of two phases at room temperature, namely gaseous and liquid phase.

A new article of interest.

Of these sterilants, the most commonly applied substances to sterile manufacturing are ethylene oxide and vapor phase Hydrogen peroxide. Ethylene oxide is used to sterilize many plastics; vapor phase hydrogen peroxide is typically used to decontaminate barrier systems (such as isolators).  

In relation to sterilization, gases are more penetrating, more uniform in concentration and less subject to variations in temperature and relative humidity than vapors. In contrast, vapors have different concentrations in each phase. Furthermore, the kill rates in the gas and liquid phase appear to be substantially different reflecting the different concentrations and available water in each phase. Thus, conventionally, gas and vapor are considered to be separate sterilization processes.in a separate chapter (gaseous sterilization is distinct from vapor sterilization because with gas, the condensation of the agent is not a consideration in the execution of these processes).

Reference:

Sandle, T. (2017) Validation Requirements for Gaseous Sterilization Using Ethylene Oxide, Journal of Validation Technology, 23 (5): 1-10

See: http://www.ivtnetwork.com/article/validation-requirements-gaseous-sterilization-using-ethylene-oxide

Posted by Dr. Tim Sandle

Sunday, 11 February 2018

Pharmaceutical Microbiology - Provides Remedial Ways to Help People


Microbiology is commonly known as the study of those organisms that cannot be seen with the naked eyes. Such organisms are termed as microorganisms as they are too small to be seen. Some of the very famous examples of microorganisms are Fungus, protists, and prokaryotes. Viruses normally are not properly known as living beings; however, they are also tested properly when it comes to microbiology. In reality, microbiology is actually a wide thing encircling virology, parasitology, bacteriology, over and above usually comprise of the investigation into the body's defense system or just immunology.

Guest post by Dr. Amol Bamane
Pharmaceutical microbiology is a further extension of microbiology. Every hospital and research labs generally includes a medical microbiology division, where one can frequently find several sorts of clinical samples, for instance, swabs, feces, urine, blood, sputum, cerebrospinal fluid, synovial fluid, and contaminated tissues etc. over here the process primarily searches for alleged pathogens and considering them, together with estimating to find out whether your identified pathogen is absolutely extremely perceptive or resistant in front of an extremely suggested drug. The result is a report identifying the infection-causing microorganism along with the type and in addition to the number of drugs that have to be given to the affected individual.

Ever since the evolution of mankind, erectile dysfunction causes and symptoms have become an indispensable part of our lives. Health problems are on the rise and with it, the pharmaceutical business. Pharmaceutical companies provide drugs or medicines that are required for medical treatments. Earlier in India, these medicines were provided by various multinational companies. However, this monopoly ended with the Indian pharmaceutical industry taking a revolutionary turn in the 1970s with the enactment of the Indian Patent Act that allowed local development of pharmaceutical industry.

The microbiology laboratory in any of the hospitals or clinics may probably also come across parasitic living beings as well as viruses. Work related to blood is frequently furthermore carried out in the laboratories, where, utilizing entire blood samples, entire blood calculations in addition to blood evaluations are completed and the conclusion is drawn based on all such things. On event, blood specimens likely to be citrated to evaluate blood clotting moments as well as coagulation factors.
  • Sub-divisions of Microbiology:
  • Microbiology is divided into many sub-disciplines as follows.
  • Cellular microbiology (combination of microbiology and cell biology)
  • Medical microbiology (study of pathogenic microbes related to diseases)
  • Environmental microbiology (study of function of microbes in natural environments)
  • Veterinary microbiology (study of microbes in veterinary medicines)
  • Evolutionary microbiology (study of evolution of microbes)
  • Industrial Microbiology ( usage of microbes for industrial purposes)
Apart from these, there are many other fields of microbial physiology, aero microbiology, microbial genetics, food microbiology, agricultural microbiology and many sub-disciplines as well.

Benefits of Microbiology:

No doubt microbes are related to human illnesses, but there are many microbes which are advantageous to human beings. Industrial fermentation which involves the production of alcohol, dairy products, and vinegar is one of the most popular ways wherein microbes are utilized. The antibiotic production also involves microbes.

Scientists and researchers have been able to produce enzymes for their usage in genetic systems. Enzymes which are biotechnologically important include Taq polymerase and even reporter genes. Microorganisms produce biopolymers which are extensively used. Microorganisms are also used for erectile dysfunction cure and medicine.

With only 1 percent of all microbes' related species being studied, the scope of microbiology is huge and its study and research are leading the field to new horizons.

Pharmaceutical Microbiology contains numerous important purposes, that are not only restricted to the hospitals but are also required in clinics. Nowadays one can easily search such associations that proffer erectile dysfunction treatment concerning remedial procedure to manufacture the product that fulfills all the fundamentals that also along with authenticated product or service assurance. Examinations cover facility validation, unprocessed materials examinations, sterilization and wrapping supports, biocompatibility, studies related to the devices that can be used again in addition to planned quality control evaluations. Such sort of tests will assist the producers of remedial mechanisms to administer to get their products to sell faster and reduce the risk.

A number of microbiologists are working in the medication industry. The medicines, other medical aids, and certain health supplements are manufactured and researched carefully in the concerned microbiology laboratories.

The businesses that are engaged in the business of supplying food include a group of microbiologists that are highly skilled and help the industry in proffering such product to the people that are properly made by adopting various quality control measures. The Foods product that will spoil speedily can be difficult to endorse, that is why microbiologists are usually there to provide the essential guidance in order to increase the production by good packaging techniques to reduce spoilage and also amplify freshness of the foodstuff.

How good bacteria control your genes


Scientists have discovered a way that bacteria in the gut can control genes in our cells. Their work shows that chemical messages from bacteria can alter chemical markers throughout the human genome. The signal chemicals are made when bacteria digest fruits and vegetables. By communicating in this way, the bacteria may help to fight infections and to prevent cancer.

This new research shows that the short chain fatty acids increase the number of chemical markers on our genes. These markers, called crotonylations, were only discovered recently and are a new addition to the chemical annotations in the genome that are collectively called epigenetic markers. The team showed that short chain fatty acids increase the number of crotonylations by shutting down a protein called HDAC2. Scientists think that changes in crotonylation can alter gene activity by turning genes on or off.

The team studied mice that had lost most of the bacteria in their gut and showed that their cells contained more of the HDAC2 protein than normal. Other research has shown that an increase in HDAC2 can be linked to an increased risk of colorectal cancer (here and here). This could mean that regulating crotonylation in the genome of gut cells is important for preventing cancer. It also highlights the important role of good bacteria and a healthy diet in this process.

See:

Rachel Fellows, Jérémy Denizot, Claudia Stellato, Alessandro Cuomo, Payal Jain, Elena Stoyanova, Szabina Balázsi, Zoltán Hajnády, Anke Liebert, Juri Kazakevych, Hector Blackburn, Renan Oliveira Corrêa, José Luís Fachi, Fabio Takeo Sato, Willian R. Ribeiro, Caroline Marcantonio Ferreira, Hélène Perée, Mariangela Spagnuolo, Raphaël Mattiuz, Csaba Matolcsi, Joana Guedes, Jonathan Clark, Marc Veldhoen, Tiziana Bonaldi, Marco Aurélio Ramirez Vinolo, Patrick Varga-Weisz. Microbiota derived short chain fatty acids promote histone crotonylation in the colon through histone deacetylasesNature Communications, 2018; 9 (1) DOI: 10.1038/s41467-017-02651-5


Posted by Dr. Tim Sandle

Saturday, 10 February 2018

Production of aromatic polyesters by E. coli strains


Systems metabolic engineers defined a novel strategy for microbial aromatic polyesters production fused with synthetic biology from renewable biomass. The team produced aromatic polyesters from Escherichia coli (E. coli) strains by applying microbial fermentation, employing direct microbial fermentation from renewable feedstock carbohydrates.

This is the first report to determine a platform strain of engineered E. coli capable of producing environmentally friendly aromatic polyesters. This engineered E. coli strain, if desired, has the potential to be used as a platform strain capable of producing various high-valued aromatic polyesters from renewable biomass. This research was published in Nature Communications on January 8.

Conventionally, aromatic polyesters boast solid strength and heat stability so that there has been a great deal of interest in fermentative production of aromatic polyesters from renewable non-food biomass, but without success.

However, aromatic polyesters are only made by feeding the cells with corresponding aromatic monomers as substrates, and have not been produced by direct fermentation from renewable feedstock carbohydrates such as glucose.

To address this issue, the team prescribed the detailed procedure for aromatic polyester production through identifying CoA-transferase that activates phenylalkanoates into their corresponding CoA derivatives. In this process, researchers employed metabolic engineering of E. coli to produce phenylalkanoates from glucose based on genome-scale metabolic flux analysis. In particular, the KAIST team made a modulation of gene expression to produce various aromatic polyesters having different monomer fractions.

The research team successfully produced aromatic polyesters, a non-natural polymer using the strategy that combines systems metabolic engineering and synthetic biology. They succeeded in biosynthesis of various kinds of aromatic polyesters through the system, thus proving the technical excellence of the environmentally friendly biosynthetic system of this research. Furthermore, his team also proved the potential of expanding the range of aromatic polyesters from renewable resources, which is expected to play an important role in the bio-plastic industry.

See:

Jung Eun Yang, Si Jae Park, Won Jun Kim, Hyeong Jun Kim, Bumjoon J. Kim, Hyuk Lee, Jihoon Shin, Sang Yup Lee. One-step fermentative production of aromatic polyesters from glucose by metabolically engineered Escherichia coli strainsNature Communications, 2018; 9 (1) DOI: 10.1038/s41467-017-02498-w

Posted by Dr. Tim Sandle

Friday, 9 February 2018

Fungi and dead trees: new insights



So far, little research has been conducted on fungi that live on dead trees, although they are vital to the forest ecology by breaking down dead wood and completing the element cycle between plants and soil. Soil biologists have now discovered that the number of fungus species inhabiting dead trees is 12 times higher than previously thought. Once trees die they are also colonized by different fungal communities depending on their species.

The study took the UFZ researchers to three areas of temperate forests in the Schorfheide-Chorin Biosphere Reserve, the Hainich National Park and the Schwa?bische Alb Biosphere Reserve, where they laid out around 300 dead tree trunks of eleven different species, each up to four metres long. The trees included seven deciduous species such as beech, oak, poplar and ash and four coniferous species: spruce, Scots pine, Douglas fir and larch. Three years later they returned to see what kind of fungal communities had established themselves in the trunks. The results were astonishing: "The diversity of fungi living in the trees was an order of magnitude greater than previously thought," says Dr Witoon Purahong, a soil ecologist based at UFZ in Halle and the first author of the study.

The researchers identified between 22 and 42 operational taxonomic units (OTUs) per trunk. OTU is a scientific term used by molecular biologists to describe organisms that can be equated with individual species due to their DNA but do not already have a species name of their own. All in all, the UFZ team identified 1,254 OTUs in the dead trunks. In a previous study, researchers found just 97 fungal species living on the same logs -- about 12 times fewer than the UFZ scientists have now discovered. Dead conifers generally had greater species diversity of fungi than most deciduous trees. The greatest diversity occurred on Douglas fir, larch and oak and the smallest amount of diversity on beech and hornbeam.

See:

Witoon Purahong, Tesfaye Wubet, Dirk Krüger, François Buscot. Molecular evidence strongly supports deadwood-inhabiting fungi exhibiting unexpected tree species preferences in temperate forests. The ISME Journal, 2017; 12 (1): 289 DOI: 10.1038/ismej.2017.177

Posted by Dr. Tim Sandle

Thursday, 8 February 2018

Ebola virus inhibited


A single enzyme. That is all the researchers behind a new study need to manipulate to prevent the feared Ebola virus from spreading. Because with the enzyme they also take away the virus' ability to copy itself and thus produce more virus particles and more infection.

The study has been published in the scientific journal Molecular Cell and was conducted by researchers from the University of Copenhagen and Phillips Universität Marburg in Germany.

'When the Ebola virus enters the human cell, its only purpose is to copy itself, fast. First it must copy all its proteins, then its genetic material. But by inhibiting a specific enzyme we rob the Ebola virus of its ability to copy itself. And that may potentially prevent an Ebola infection from spreading', says Professor Jakob Nilsson from the Novo Nordisk Foundation Center for Protein Research.

A few years ago the Ebola virus ravaged West Africa, where thousands of people died from the extremely infectious Ebola infection. Once you are infected, all you can do is hope that your own immune system is able to kill the infection. Because there is currently no available treatment.

However, the researchers behind the new study have found what is called a new host factor for Ebola virus. It can be described as a small part of the host's -- for example the human body's -- own cells, which the Ebola virus uses to copy itself and produce more infection.

The virus uses the host factor enzyme PP2A-B56 to start producing proteins. So if the researchers switch off PP2A-B56, the virus' ability to copy itself and produce more infection is never 'switched on'.

See:

Thomas Kruse, Nadine Biedenkopf, Emil Peter Thrane Hertz, Erik Dietzel, Gertrud Stalmann, Blanca López-Méndez, Norman E. Davey, Jakob Nilsson, Stephan Becker. The Ebola Virus Nucleoprotein Recruits the Host PP2A-B56 Phosphatase to Activate Transcriptional Support Activity of VP30Molecular Cell, 2017; DOI: 10.1016/j.molcel.2017.11.034


Posted by Dr. Tim Sandle

Wednesday, 7 February 2018

Monocyte Activation Test (MAT) - The in vitro test for pyrogen detection – White paper



Monocyte Activation Test (MAT)
The in vitro test for pyrogen detection

Pyrogens…a hot story

Adverse reactions to parenteral preparations have been described as early as the late 19th century, frequently termed “injection fever”. The first fever causing agents, “pyrogens”, were identified in 1912 by Hort and Penfold, who were also the first to design a pyrogen test based on injection of material into rabbits. At that time, the pyrogenic agent was identified as endotoxins included in preparations of gram-negative bacteria. Interestingly, it was shown that live and dead microorganisms presented the same pyrogenic potential.

In the following years, it became more and more clear that sterility is not necessarily equal to apyrogenicity, which led to the inclusion of a pyrogen test in the 12th edition of the United States Pharmacopoeia (USP) in 1942.

Due to their stability, endotoxins can be very difficult to remove by classical bactericidal procedures such as heating or filtration. This made control of the whole production process necessary, especially for the water used, as this water was frequently found as source of pyrogenic contaminations.

The high number of pyrogen tests on rabbits and the variable sensitivity of that test system (e.g. by development of pyrogen tolerance in rabbits after repeated injections) made development of alternative tests necessary. The first and most successful of these new tests was the bacterial endotoxin test based on the lysate of amoebocytes from the blood of horseshoe crabs, which became commercially available in the 1970s and has been widely used as a replacement for the rabbit pyrogen test.

Today's qualified water systems no longer present such a high risk of endotoxin contamination, with more than 99% of our tests for various production sites showing contamination of much less than the specification of 0.25 EU/mL.

On the other hand, quality control for the presence of pyrogens is getting more and more complicated, as production processes (e.g. biotechnology and cell therapy products) bring new risks of various contaminants (i.e Non-Endotoxin Pyrogens) entering the final product, like viruses from animal-based raw materials or gram-negative bacteria from contaminations. Non-Endotoxin Pyrogens (NEPs) are undetectable by the bacterial endotoxin test,
and there is therefore a risk of overlooking a NEP contamination.

In 2016, due to the increase in production of more and more complex products, the general chapter for bacterial endotoxins testing in the European Pharmacopoeia (chapter 5.1.10) introduced the necessity for an evaluation of the product, production process and raw materials with respect to the risk for pyrogens that are non-detectable by the bacterial endotoxin test (ie Non-Endotoxin Pyrogens).

In this context, the in vitro pyrogen test based on human cells offers a valuable alternative to the rabbit pyrogen test. Since2010, the Monocyte Activation Test has been described as a compendial method for Pyrogen Detection in the European Pharmacopeia (chapter 2.6.30).

Test comparison

Both RPT and LAL tests are animal-based methods. LAL cannot can adequately detect the full spectrum of Pyrogens. Moreover, such tests cannot be used on several pharmaceutical products or for the testing of solid materials such as medical devices.





White-nose syndrome fungus may have an Achilles' heel


In the course of genomic analyses of the fungus behind white-nose-syndrome, a devastating disease that has killed millions of bats in North America, US Forest Service scientists discovered something very surprising: brief exposure to UV-light kills Pseudogymnoascus destructans.

In the course of genomic analyses of P. destructans, a team of scientists from the U.S. Forest Service, U.S. Department of Agriculture and the University of New Hampshire found that the fungus is highly sensitive to UV light. P. destructans can only infect bats during hibernation because it has a strict temperature growth range of about 39-68 degrees Fahrenheit. However, treating bats for the disease during hibernation is challenging, so any weakness of the fungus may be good news to managers trying to develop treatment strategies.

In a study published on Jan. 2 in the journal Nature Communications titled "Extreme sensitivity to ultra-violet light in the fungal pathogen causing white-nose syndrome of bats," the research team suggests that P. destructans is likely a true fungal pathogen of bats that evolved alongside bat species in Europe and Asia for millions of years, allowing Eurasian bats to develop defenses against it. In the course of comparing P. destructans to six closely related non-pathogenic fungi, researchers discovered that P. destructans is unable to repair DNA damage caused by UV light, which could lead to novel treatments for the disease. The study was funded by the U.S. Fish and Wildlife Service.

See:

Jonathan M. Palmer, Kevin P. Drees, Jeffrey T. Foster, Daniel L. Lindner. Extreme sensitivity to ultraviolet light in the fungal pathogen causing white-nose syndrome of batsNature Communications, 2018; 9 (1) DOI: 10.1038/s41467-017-02441-z

Posted by Dr. Tim Sandle

Tuesday, 6 February 2018

Pharmig News #70


The 70th edition of Pharmig News has been published.

In the new issue:
  • Report from the Royal Society of Microbiology, by Julie Roberts.
  • Report of the Pharmig 25th Anniversary Conference, including reviews of the different presentations.
  • Hot topics in pharmaceutical microbiology.
  • Review of Pharmig’s Adriatic Region conference.
  • Pharmig’s round-up of regulatory news.
  • And more….
Copies have been sent to Pharmig member companies. If you are interested in reading a copy or finding out more about Pharmig, please contact Pharmig at: info@pharmig.org.uk



Posted by Dr. Tim Sandle

Monday, 5 February 2018

Revisions made to European Pharmacopoeia sterilisation chapters



Methods of sterilisation and the assessment of sterilisation using biological indicators represent important areas of regulatory concern. The two applicable guidance chapters in the European Pharmacopeia have undergone revisions. This article assesses the main changes. Notably these include reference to the production of sterile products through aseptic processing for the first time; and changes to the assessment of the suitability of biological indicators, including a new recommendation for supplier auditing.

Tim Sandle has written a new article for Clean Air and Containment Review. The reference is:

Sandle, T. (2017) Revisions made to European Pharmacopoeia sterilisation chapters, Clean Air and Containment Review, Issue 32, pp18-20

Posted by Dr. Tim Sandle

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