Saturday, 20 September 2014

Sample testing advice for Ebola virus

As part of Public Health England’s Ebola virus disease: clinical management and guidance there is a section on frequently asked questions on laboratory testing of samples from patients with possible Ebola virus disease.

Guidance on the risk assessment and management of viral haemorrhagic fevers (VHF), including Ebola, was updated by the Advisory Committee on Dangerous Pathogens (ACDP) in August 2014 and is the principal source of guidance for clinical staff managing patients.

For further information see: Ebola: sample testing advice for laboratory staff.

Posted by Tim Sandle

Friday, 19 September 2014

Pharmacy Compounding of Human Drug Products Under Section 503A of the Federal Food, Drug, and Cosmetic Act


This guidance announces FDA’s intention with regard to enforcement of section 503A FD&C Act to regulate entities that compound drugs, now that section 503A has been amended by Congress to remove the advertising and solicitation provisions that were held unconstitutional by the U.S. Supreme Court in 2002. Several parts of section 503A require rulemaking and consultation with a Pharmacy Compounding Advisory Committee to implement. This guidance explains how the provisions will be applied pending those consultations and rulemaking. This guidance also describes some of the possible enforcement actions FDA can bring against individuals or firms that compound drugs in violation of the FD&C Act.

See: FDA

Posted by Tim Sandle

Leading pharmacists debate hospital infections

This week the leading pharmacists in the U.K. congregated in Warwick for the NHS QA Symposium. A number of measures to protect patients from infection were discussed, and Digital Journal was in attendance.
Most of the U.K.’s health provision is delivered through the National Health Service (NHS). Among the care provided is the manufacture of small batches of medicines through specialist pharmacy units. Those involved in this process include technicians, medics, pharmacists and microbiologists. The main principle by which the medicines are produced is Quality assurance (QA).
Once a year, managers, specialists and scientists gather to discuss the latest technologies and best practices at the QA Symposium.
One of the running themes was about making error free medicines. A session on this subject was led by Professor Anna Guha, from Royal Liverpool and Broadgreen University Hospitals NHS Trust. The talk focused on getting procedures right and understanding different learning styles. Control and procedures featured in several talks, including the appropriate design of software. One presentation by Peter Rhodes looked at the ideal software for hospital compounding.
A lively debate took place about “vial sharing”. This is whether a vial of drug should be single-use or shared between patients. Although most drugs come in single-dose vials (SDVs) they typically contain more drug than one dose. On this basis, can the same vial be used for more than one patient? The argument for vial sharing is in relation to cost savings. The argument against centered on risks to patient safety, especially the risk of contracting a hospital acquire infection. The issue that everyone seemed to agree on is that the “leftovers” from two different vials should not be pooled together, mixed, and then administered to a patient.
In a presentation on pharmaceutical microbiology, Dr. Tim Sandle discussed the need to find out exactly what types of microorganisms are present in the environment and then to apply this knowledge to select the best disinfectants and biocides that can kill these microorganisms so they do not remain in the areas where medicines are prepared. Following this, Nicola Swift explained why characterizing microorganisms was important: “accurate identification is of great importance”, she explained, before outlining some advances in rapid and more accurate methods.
In a related area, there were several stands and posters providing information about hospital acquired infections. Here a strong emphasis was placed on healthcare professionals washing their hands and using effective hand sanitizers, as a means to lower the risk of infection to patients.
The thorny issue of outsourcing was brought to the table by pharmacist Richard Bateman. This relates to the issue of the extent that health provision in the U.K. is subject to privatisation. Bateman’s presentation was focused on maintaining quality standards. Talking to many of the delegates, the vast majority who expressed an opinion were concerned about privatization of the health service, especially with outside companies coming in and having lower quality standards.
Overall the conference raised and debated a number of contemporary and important issues relating to healthcare and the preparation of medicines.

Posted by Tim Sandle

AC5 Surgical Hemostatic Device(TM) in Animals on Blood Thinner

Arch Therapeutics, Inc, developer of the AC5 Surgical Hemostatic Device™ for potential use in bleeding during surgery, has announced positive data in an initial preclinical study assessing the use of its AC5 Surgical Hemostatic Device™ in animals receiving an anticoagulant medication (i.e. blood thinner).
In the first preclinical assessment of AC5 in the presence of an anticoagulant, AC5 quickly stopped brisk bleeding from a 4mm diameter biopsy surgical site created in the livers of nine rats that had been treated with a clinically relevant dose of the drug heparin. Heparin is an anticoagulant commonly given to patients to prevent blood clots during and after surgical procedures and in other circumstances. The study demonstrated that application of AC5 in this commonly used animal model of brisk bleeding was able to stop blood loss from the liver in less than 30 seconds and to achieve hemostasis. Time to hemostasis (TTH) after creation of the liver wounds in heparin-treated animals was comparable to TTH after creation of liver wounds in normal, non-anticoagulated animals.
"There is a great need to continue to develop novel hemostatic agents and sealants that are efficacious in surgical and trauma patients," said Cambridge, MA-based surgeon Steven Schwaitzberg, MD, Professor of Surgery at Harvard Medical School and advisor to Arch Therapeutics. "In particular, the need is greatest in those patients whose underlying coagulation cascade or platelet status is abnormal, whether due to concurrent antithrombotic therapy or an underlying disease. Increasing numbers of patients are on anticoagulant or antiplatelet medications. This can present a challenge to surgeons and other interventionalists when procedures are needed on these patients. These initial findings on the activity of AC5 in this setting are very encouraging and may lead to significant benefit in the future."
Terrence W. Norchi, MD, President and CEO of Arch Therapeutics, said, "We are excited by the data from this early study. This is the first of several planned preclinical studies intended to assess the utility of AC5 in animals on antiplatelet or anticoagulant therapy, commonly referred to as blood thinners. A significant portion of the population takes blood thinners to prevent a range of life-threatening conditions. Patients on these medications are at increased risk of bleeding and their management during surgical and other procedures requires extra care. We hope to demonstrate that AC5 is effective regardless of a patient's underlying platelet or bleeding status, whether due to prescribed medications or an underlying condition such as hemophilia, in order to offer surgeons and patients an even better tool to control bleeding."

For further details see: Arch Therapeutics 

Posted by Tim Sandle

Thursday, 18 September 2014

Decoding dengue

Scientists have discovered a new pathway the dengue virus takes to suppress the human immune system. This new knowledge deepens our understanding of the virus and could contribute to the development of more effective therapeutics.

Researchers from the Program in Emerging Infectious Diseases (EID) at Duke-NUS Graduate Medical School Singapore (Duke-NUS) have discovered a new way that dengue virus-2 (DENV-2) uses to evade the human defense system. Typically, when a virus enters the body and infects cells, it induces the production and release of interferons (IFNs), which are proteins that raise the bodies' anti-viral defense mechanisms.

The dengue virus enters the cell and produces large quantities of a non-coding, highly-structured viral RNA termed sfRNA, which is part of the genetic material of the dengue virus.

For further details, see Biology News

Posted by Tim Sandle

Wednesday, 17 September 2014

Studying intestinal microbiota

Analyzing the global genome, or the metagenome of the intestinal microbiota, has been advanced through a new approach. This method simplifies microbiome analysis. Through this scientists have been able to sequence and assemble the complete genome of 238 intestinal bacteria, 75% of which were previously unknown.

It is estimated that 100,000 billion bacteria populate the gut of each individual (or 10 to 100 times more than the number of cells in the human body), and their diversity is considerable, estimated to around a thousand different bacterial species in the intestinal human metagenome. These bacteria have become major factors in understanding certain diseases such as obesity, type 2 diabetes, or Crohn's disease.


Researchers from INRA, together with teams from CEA (Genoscope), CNRS and Université d'Evry in France, and scientists from other countries, have developed a new method that can markedly facilitate analysis of the gut metagenome, while at the same time improving the quality of the data obtained. To achieve this, they based themselves on a simple hypothesis:

•Within a bacterial species harboured by the gut of an individual, the abundance of genes remains constant, since every bacterium of a same species have the same genes
•However, the relative abundance of these different species can vary markedly between individuals, from 10-fold to 1000-fold, so that of course the abundance of genes harboured by an individual varies to the same extent.
•By measuring the abundance of bacterial genes in different individuals, it would therefore be possible to group the genes of a specific bacterial species, because their abundance is the same in a particular individual but differs between individuals.

An analysis of 396 stool samples from Danish and Spanish individuals allowed the researchers to cluster these millions of genes into 7381 co-abundance groups of genes. Approximately 10% of these groups (741) corresponded to bacterial species referred to as metagenomic species (MGS); the others corresponded to bacterial viruses (848 bacteriophages were discovered), plasmids (circular, bacterial DNA fragments) or genes which protected bacteria from viral attack (known as CRISPR sequences). 85% of these MGS constituted unknown bacteria species (or ~630 species).

Using this new approach, the researchers succeeded in reconstituting the complete genome of 238 of these unknown species, without prior culture of these bacteria. Living without oxygen, in an environment that is difficult to characterise and reproduce, most of these gut bacteria cannot be cultured in the laboratory. And until now, analysis of the metagenome was based on comparing the genes detected in a sample with those listed in catalogues of genes from bacteria that were known and could be cultivated in a laboratory (or 15% of gut bacteria), so that it was impossible to assign genes to non-cultivable bacteria.

The authors also demonstrated more than 800 dependent relationships within the 7381 gene co-abundance groups; this was the case, for example, of phages which require the presence of a bacterium to survive. These dependent relationships thus enable a clearer understanding of the survival mechanisms of a micro-organism in its ecosystem. It is also the first time that an analysis has clarified the relationships between different biological entities in the gut microbiota, which will facilitate their detection, isolation and culture.

This study throws a detailed light on microbial communities in humans. The method thus developed enables considerably simpler analysis of genes in the gut microbiota; it is now possible to study just a few thousand genetic elements, or hundreds of species, rather than the millions of genes that make up the metagenome. The method also markedly improves the reliability and accuracy of statistical analyses.

The research findings have been published in Nature Biotechnology. The reference is:

H Bjørn Nielsen, S Dusko Ehrlich et al. Identification and assembly of genomes and genetic elements in complex metagenomic samples without using reference genomes. Nature Biotechnology, 2014; DOI: 10.1038/nbt.2939

 Posted by Tim Sandle

Tuesday, 16 September 2014

Microbiome science needs a healthy dose of scepticism



To guard against hype, those interpreting research on the body's microscopic communities should ask five questions, says William P. Hanage writing in Nature.

The questions are: 
  • Can experiments detect differences that matter?
  • Does the study show causation or just correlation?
  • What is the mechanism? A return to a reductionist approach is essential if we are to pinpoint both whether the microbiome affects human health, and exactly how it does so.
  • How much do experiments reflect reality?
  • Could anything else explain the results? There are good reasons to think that bacteria influence us in a host of ways. But there are many other — possibly more important — influences, such as diet.
At the end of the commentary, Hanage concludes: “The hype surrounding microbiome research is dangerous, for individuals who might make ill-informed decisions, and for the scientific enterprise, which needs to develop better experimental methods to generate hypotheses and evaluate conclusions. Funding agencies must not let their priorities be distorted by the buzz around the field, but look dispassionately at the data. Press officers must stop exaggerating results, and journalists must stop swallowing them whole. In pre-scientific times when something happened that people did not understand, they blamed it on spirits. We must resist the urge to transform our microbial passengers into modern-day phantoms.”

To read the article, go to Nature.

Posted by Tim Sandle

Monday, 15 September 2014

The Lean Laboratory and Environmental Monitoring

Running an efficient laboratory requires the laboratory manager to balance the important aspects of compliance alongside the need to manage a budget and often scarce resources. Efficiencies can be generated by applying the philosophy of the “lean laboratory.” The “lean labs” approach focuses on cost control, improving sample throughput, and reviewing whether each sample tested adds value or produces meaningful information. Addressing this latter point can sometimes result in sample reduction.

In terms of the volume of samples processed throughout quality control laboratories, it is invariably the microbiology laboratory that processes the highest number of samples. Aside from raw materials and intermediate and finished product samples, this is a consequence of the sampling of water systems and the environmental monitoring of cleanrooms. Large facilities, with many cleanrooms, can generate tens of thousands of environmental monitoring samples each year.

To explore the lean labs concept, drawing on the context of environmental monitoring, Tim Sandle has written a paper for Journal of Validation Technology.

The paper introduces the concept of the lean laboratory and then illustrates how the principle of lean labs can be applied. This illustration is through a case-study that outlines an approach for the removal of samples from the environmental monitoring program for areas of a lower classification. The emphasis is upon removing samples in such a way that the absence of the data does not detract from the overall assessment regarding the status of the cleanroom.


The reference is:

Sandle, T. (2014) The Lean Laboratory and Its Application for the Review of Environmental Monitoring Samples, Journal of Validation Technology, Vol. 20, Issue 2, Jun 2014. Published on-line.

For further details see: IVT or contact Tim Sandle

Posted by Tim Sandle

Sunday, 14 September 2014

NHS Pharmaceutical QA Service Annual Symposium, 16th-17th September 2014


Organised by the NHS Pharmaceutical Quality Assurance Committee, the annual event brings together key pharmaceutical staff from across the UK. The 2014 programme offers a variety of presentations and workshops to educate and update delegates on current issues within the pharmacy industry, with a focus on "Optimising Patient Care Safely".

The event is taking place at Chesford Grange Hotel, Kenilworth on 16th – 17th September 2014.

For further details see: NHS QA

Posted by Tim Sandle

How do immune responses control microbial gene expression?

Commensal microbiota provide protection from bacterial infections. However, when the body's control mechanisms are disrupted, microbial gene expression may be altered, ultimately changing bacterial behavior and localization.

High levels of bacterial flagellar protein are associated with gut mucosal barrier breakdown. Using mice deficient in Toll-like receptor 5 (TLR5), researchers found that innate and adaptive immunity interact to modulate the microbiome’s production of flagella, and the flagella of gut commensal bacteria stimulate TLR5 (Cullender, T.C. et al. (2013) Cell Host Microbe 14, 571).

Posted by Tim Sandle

Saturday, 13 September 2014

World Sepsis Day

September 13 is World Sepsis Day




World Sepsis Day is an international day of action and awareness-raising, supported by organisations around the world. It is coordinated internationally by the Global Sepsis Alliance, a collaborative group of non-profit organisations.

Sepsis is a life threatening condition that arises when the body’s response to an infection injures its own tissues and organs. It can be caused by something as simple as a cut or insect bite, or an infection like pneumonia. It is also a risk following surgery, or for women who have just given birth. It causes a range of diseases and deaths worldwide.
For this reason, World Sepsis Day is promoted on September 13 each year in order to raise awareness. It is is an initiative of the Global Sepsis Alliance and its founding members, all of whom are non-profit organizations.
According to the campaign organization: "Every few seconds someone dies of sepsis. Prevent it. Spot it. Treat it – beat it. For that we campaign to reduce sepsis cases by 20% by 2020".
To give an indication of the extend of the problem, in the U.S. Dr. Liu, from the Kaiser Permanente Northern California, has determined that with U.S. hospitals:

a) Sepsis contributed to 1 in every 2 to 3 deaths, and most of these patients had sepsis at admission.
b) Patients with sepsis, normal blood pressure, and measured lactate levels of less than 4 mmol/L (n = 15 095) comprised 55.9% (95%CI, 53.6%-58.1%) of sepsis deaths. Surprisingly, patients with initially less severe sepsis made up the majority of sepsis deaths. The majority of individuals who died with sepsis presented to the hospital with the illness. This contradicts the belief of policymakers and some healthcare authorities that sepsis results primarily from hospital-acquired infections.
One way of dealing with problems of sepsis is in developing appropriate diagnostic tools. According to Pharmaceutical Microbiology: "Diagnostics play a significant role in identifying sepsis in a patient. Surviving Sepsis Campaign guidelines recommend obtaining blood cultures - the gold standard - for recovering the pathogen causing infection before antimicrobial therapy is initiated. Accurate and fast results help physicians to determine the appropriate antimicrobial therapy to cure the patient."
Whilst diagnosis is important, a lack of international agreement on appropriate guidelines is arguably hampering efforts to address sepsis on a global scale. Professor Sandra Peake, of The Queen Elizabeth Hospital in England, says that guidelines continue to be debated among clinicians and researchers: "In 2004, international guidelines were introduced for the resuscitation of patients with sepsis, but even now we see variation in practices within and between countries on treatment approaches.
"The debate will continue internationally for some time yet, but in reality the mainstay of treatment for patients with sepsis is antibiotics, intravenous fluids, drugs to support the heart and maintain the blood pressure, and surgery, if needed, on an affected area. Whether or not there is one specific or uniform resuscitation approach remains to be seen."
Therefore, it remains an issue that treatment practices for patients hospitalized with sepsis will continue to vary because of individual differences between hospitals and countries.
You can sign up to support World Sepsis Day here.

Posted by Tim Sandle

Friday, 12 September 2014

First crystal structure of the C. difficile surface protein

Clostridium difficile is a major problem as an aetiological agent for antibiotic-associated diarrhea. The mechanism by which the bacterium colonizes the gut during infection is poorly understood, but undoubtedly involves a myriad of components present on the bacterial surface. This study provides some insights that may help in developing a new type of drug to treat the infection.

Researchers reported the first crystal structure of the C. difficile surface protein Cwp84. This cysteine protease enzyme is found on the surface of the bacterium and assists with production of the microbe's surface-layer, which is likely to play an essential step in the colonisation of the gut. The enzyme cleaves a single polypeptide (surface-layer protein A; SlpA) into low- and high-molecular-weight subunits.

Scientists have identified three critical regions in a mutant of the enzyme that could represent novel targets for drugs to attack C. difficile by blocking maturation of its surface layer during colonisation.

For further details, see:

William J. Bradshaw, Jonathan M. Kirby, Nethaji Thiyagarajan, Christopher J. Chambers, Abigail H. Davies, April K. Roberts, Clifford C. Shone, K. Ravi Acharya. The structure of the cysteine protease and lectin-like domains of Cwp84, a surface layer-associated protein fromClostridium difficile. Acta Crystallographica Section D Biological Crystallography, 2014; 70 (7): 1983 DOI: 10.1107/S1399004714009997

Posted by Tim Sandle

Thursday, 11 September 2014

'Revolution' biomotor discovered in many bacteria

Scientists at the University of Kentucky, led by nano-biotechnologist Peixuan Guo, have made an important discovery into the operation of biomotors, the molecular machines used by viruses and bacteria in the packaging of DNA. Biomotors function similarly to mechanical motors but on a nano-scale.

The researchers have reported on the discovery of a new, third class of biomotor, unique in that it uses a "revolution without rotation" mechanism. Rotation is the turning of an object around its own axle, as the Earth does every 24 hours. Revolution is the turning of an object around a second object, as the Earth does around the sun.

The reference for the research is:

Cell & Bioscience 2014, June, 4:30: www.cellandbioscience.com/content/4/1/30



 Posted by Tim Sandle

Wednesday, 10 September 2014

Candida glabrata

A group of researchers at the Max F. Perutz Laboratories has created one of the three world's largest gene libraries for the Candida glabrata yeast, which is harmful to humans. Molecular analysis of the Candida glabrata fungus mutations led to the discovery of 28 new genes that are partly responsible for the yeast's tolerance of common drugs.

The working group led by Karl Kuchler at the Max F. Perutz Laboratories (MFPL) - a research and training centre run jointly between the University of Vienna and the Medical University of Vienna at the Vienna Biocenter Campus - coordinated an international study cooperation aimed at researching new tolerance and virulence genes in Candida glabrata. During this process, genetic methods were used to generate one of the three world's largest libraries of "knock-out fungi". More than 600 fungus mutations were created from which a single gene was specifically removed.

The findings represent a new step in the discovery and characterisation of Candida glabrata resistance genes, laying the foundations for the development of new anti-fungal medications.

For reference see: "Systematic Phenotyping of a Large-Scale Candida glabrata Deletion Collection Reveals Novel Antifungal Tolerance Genes" – Tobias Schwarzmüller, et al

PLoS Pathogens 10: e1004211. DOI: 10.1371/journal.ppat.1004211

Posted by Tim Sandle

Tuesday, 9 September 2014

Challenges of GMP and GXP in India

Pharmig are hosting the following free-to-view presentation 'Challenges of GMP and GXP in India'.

GMP and regulatory issues affecting pharmaceuticals and healthcare in India. The presentation covers Good Manufacturing Practices, and includes national and global (e.g. FDA) requirements.

The presentation has been prepared by Nasir  Ansari  of  Piramal  Enterprises  Limited  .


Posted by Tim Sandle