Saturday 31 January 2015

Good practices for Pharmaceutical Microbiology Laboratories: A review of the WHO Guidance

A copy of Tim Sandle’ article ‘Good practices for Pharmaceutical Microbiology Laboratories A review of the WHO Guidance’ has been published on line by the site GMP Logfile. The article can be read on-line.

Here is an extract:

“The document is useful, however, in stating the need for supervision and for authorised personnel for the release of results from the laboratory. The document also outlines some of the basic training required for staff including aseptic technique, serial dilutions, working with hazardous cultures, and colony counting. Although there is not a great deal of detail, the section is important because common agreement, nationally or internationally, on microbiology laboratory training is lacking.

The second substantive section concerns laboratory premises. In this part reference is made to the need for dedicated areas for laboratory equipment, layout for operations, the separation of laboratories from production areas, and controlled access to microbiology areas. Reference is also made to cleaning and disinfection of the laboratory, spillage policy and hand sanitisation.”

To view, see GMP.

Posted by Tim Sandle

Friday 30 January 2015

Ethylene Oxide Test Gas on the Resistance of Biological Indicators

On  Dec. 31, 2014, the United States Environmental Protection Agency Clean Air Act prohibited the sale and use of HCFC-based (hydrochlorofluorocarbon) products in the US, and that will include Oxyfume ethylene oxide (EtO) sterilant blends such as Oxyfume 2000, which consists of 8.6% EtO and 91.4% HCFC-124. (Oxyfume is a registered trademark owned by Honeywell International.) This requirement means that all BI manufacturers will have to move to 100% EtO as the test gas for determining the resistance performance of EtO BIs by the end of 2014. Currently, Oxyfume 2000 is often used by BI manufacturers for assessing BI EtO D-value label claims, so it is in the best interest of the BI community (manufacturers and end users) to assess the potential effects of this change.

To explore the implications of this change, Anthony M. Piotrkowski, Kellie A. Matzinger, Garrett Krushefski, Craig A. Wallace have written a review for Pharm. Tech. the article can be viewed here.

 Posted by Tim Sandle

Thursday 29 January 2015

Microbiome research

Recently, scientists pegged microbes as important players in several aspects of human health. Scientists have long known that people are a host for a range of different microbes; however, figuring out the myriad roles of bacteria in the body is still a work in progress. Science News has an interesting overview of current research.

See: Science News

 Posted by Tim Sandle

Wednesday 28 January 2015

Environmental Monitoring: a Comprehensive Handbook

A new book of interest:

This is volume 7 of the Environmental Monitoring Handbook series. Each volume of this series discusses different aspects of environmental monitoring. The appendix describes the various topics and authors that can be found in volumes 1 through 6 of this series… There is a wealth of useful information that you can use in establishing, maintaining and updating your environmental monitoring program!

For followers of this blog, Tim Sandle has written a chapter. The chapter reference is:

Sandle, T. (2015) Cleanroom Design. In Moldenhauer, J. (Ed.) Environmental Monitoring: a Comprehensive Handbook, Volume 7, pp3-28

The book is available from PDA/DHI

Posted by Tim Sandle

Tuesday 27 January 2015

The Risk of Bacillus cereus to Pharmaceutical Manufacturing

A review of warning letters U.S. Food and Drug Administration indicates that contamination events associated with Bacillus species represents a relatively high proportion of microbiological related citations. During the period March 2013 to August 2014, 7 warning letters relating to contamination from spore-forming bacteria were issued. Extending the review back to 2007, it is noteworthy that the most common microorganism associated with contamination is Bacillus cereus.

This article considers some of the inspectorate findings relating to Bacillus contamination and goes on to consider the implications for the control of pharmaceutical products manufacture that arise from these regulatory observations. The discussion has a focus on Bacillus cereus, given its relative ubiquity, and extends to the general risks that arise from spore-forming microorganisms and the risk-mitigating actions that can be taken.

This is the basis of a new article by Tim Sandle for American Pharmaceutical Review. The article is available to be viewed in full on-line. Go to: APR.

Sandle, T. (2014) The Risk of Bacillus cereus to Pharmaceutical Manufacturing, American Pharmaceutical Review, 17 (6): pp-pp

Monday 26 January 2015

Susceptibility Testing for MRSA with the PCR assay

A new article of interest:

Methicillin-resistant Staphylococcus aureus (MRSA) is a multi-drug resistant pathogen, which is responsible for increasing cases of serious diseases, including life-threatening diseases and nosocomial and community-acquired infections. Laboratory identification of MRSA is crucial and essential both for initiation of appropriate antimicrobial therapies and for effective infection control strategies that are designed to limit the spread of MRSA. In spite of the availability of commercial kits for MRSA detection in the market, the Clinical and Laboratory Standards Institute (CLSI) recommends the use of phenotypic methods, such as the disk diffusion method with oxacillin or with cefoxitin, as well as a serial dilution method with oxacillin. Nevertheless, some studies have shown that results obtained with such phenotypic methods are controversial. The aim of the study described in this paper was to comparatively evaluate the traditional susceptibility testing for MRSA with PCR as the gold standard assay. Analysis of collection (n = 68) isolates of Staphylococcus aureus revealed that the serial dilution method with oxacillin possessed the highest sensitivity (at 100%). In contrast, the disk diffusion methods with oxacillin and cefoxitin showed lower sensitivity (95.83%, 95% CI (78.81% - 99.30%)). Furthermore, the borderline value of zone inhibition diameters for cefoxitin might be considered as a risk, and they may give false-susceptible result.

The reference is:

Sandle, T., Azizov, I., Babenko, D., Lavrinenko, A., Chesca, A. (2014) Comparative Evaluation of Traditional Susceptibility Testing for MRSA with the PCR Approach, Advances in Microbiology, 4, 1204-1211

For a copy, please contact Tim Sandle

Posted by Tim Sandle

Sunday 25 January 2015

Deep sea oil reservoirs contain social bacteria

New research, led by scientists at Dartmouth College and University of Oslo, has shown that microbes living in the ‘deep biosphere’ – the sediment found far beneath the ocean floor – have been swapping genes for aeons. The work, published in the ISME Journal, compared heat-loving bacteria living in two environments: subsurface oil reservoirs and near hot water vents on the ocean floor. By studying the DNA from the two sample groups, the researchers showed that genes found within microbes living in the subterranean oil were also found on the ocean floor group, suggesting that bacteria have moved between the two. As yet, the mechanisms behind this ‘gene flow’ are unknown.

See: ISME Journal

Posted by Tim Sandle

Saturday 24 January 2015

Dengue fever vaccine close to development

The antibodies could be used to treat dengue fever or develop a vaccine that works against all four strains of virus, according to The Guardian.

A new class of antibody found in the blood of patients with dengue fever has boosted hopes for a vaccine against the virus, which debilitates millions and kills tens of thousands each year.

Cases of dengue fever have soared in the past 50 years to nearly 100 million a year as improved transport and urbanisation have brought more people into contact with the mosquito-borne virus.

While dengue infection often causes mild to high fever and lasts only a week or so, some patients develop dengue haemorrhagic fever, which is far more serious and kills about 22,000 people a year, many of them children.

The researchers spotted the new group of antibodies while they were studying blood drawn from patients who picked up dengue infections in south-east Asia.

They found that about a third of the immune reaction launched by each patient came from a new class of antibodies. Instead of latching on to a single protein on the virus surface – as usually happens – the new group of antibodies latches on to a molecular bridge that joins two virus proteins together.

When antibodies bind to viruses, they make them targets for attack from the wider immune system.

In tests are described in the journal Nature Immunology.
Posted by Tim Sandle

Friday 23 January 2015

FDA Issues (Draft) Guidance on 510(k) Transfers

FDA has always allowed companies to transfer 510(k) clearances without obtaining new clearance. FDA allows allows a 510(k) holder to transfer clearance without a new 510(k) clearance (absent a significant modification of the device). There may be only one 510(k) holder at a time. The process has never been entirely clear.

Now the Agency has issued draft guidanbce in the form of Q & A.

For details see: FDA.

For a good blog on the subject, see the FDA Law Blog.

Posted by Tim Sandle

Thursday 22 January 2015

Waters could support isolated life

A team of scientists has mapped the location of hydrogen-rich waters found trapped kilometers beneath Earth's surface in rock fractures in Canada, South Africa and Scandinavia. Common in Precambrian Shield rocks -- the oldest rocks on Earth -- the ancient waters have a chemistry similar to that found near deep sea vents, suggesting these waters can support microbes living in isolation from the surface.

To find out more see:

Barbara Sherwood Lollar, T. C. Onstott, G. Lacrampe-Couloume, C. J. Ballentine. The contribution of the Precambrian continental lithosphere to global H2 production. Nature, 2014; 516 (7531): 379 DOI: 10.1038/nature14017

Posted by Tim Sandle

Wednesday 21 January 2015

Iron enhanced protein fights pathogens

Meningitis-causing bacteria exerted strong evolutionary pressure on an iron-binding protein in primates, a study shows.

Damaging toxins and antibody-epitope interactions are frequently the battlegrounds of host-pathogen arms races, but a new study suggests a role for a different, so-called “nutritional” immune system. In primates, the iron-carrying protein transferrin likely evolved to inhibit meningitis-causing bacteria from scavenging iron, an essential but limited nutrient, researchers reported in Science.

In light of rising antibiotic resistance, medics have been toying with providing patients with more transferrin to help to fight in infections.

Posted by Tim Sandle

Tuesday 20 January 2015

LAL may be unsuitable for detecting endotoxin in blood

A new article of interest:

“The Limulus Amebocyte Lysate assay may be unsuitable for detecting endotoxin in blood of healthy female subjects” by Anne Gnauck and colleagues from Institute of Food, Nutrition and Human Health, Massey University, New Zealand. The article is published in the Journal of Immunological Methods (doi:10.1016/j.jim.2014.11.010).

The abstract reads:

We examined the factors that may influence the outcome of the Limulus Amebocyte Lysate (LAL) assay, when it is used for quantifying Gram-negative bacterial endotoxin, also referred to as lipopolysaccharide (LPS), in samples of human blood. We found that the method recommended by the manufacturers, based on the reaction time, was inaccurate with any type of serum samples due to the slowing of the initial phase of reaction, likely by serum proteins. We describe an alternative method that is more accurate for use with heated serum samples. Further, we found that components of fresh serum irreversibly sequester endotoxin but that this action may be largely prevented by dilution and heating, but only if this occurs prior to the addition of endotoxin. The tests also indicated that a number of types of proprietary plastic vacutainers appeared to contain significant amounts of endotoxin. However, even when appropriate blood collection containers and calculation methods were used, the levels of endotoxin in serum samples detected by LAL assay were unlikely to reflect the total quantities of endotoxin in that sample and more likely to reflect the capacity of a given serum sample to sequester endotoxin.

Posted by Tim Sandle

Monday 19 January 2015

New book - Cleanroom Microbiology

Cleanroom Microbiology is a new book written by international experts Tim Sandle (Head of Microbiology at BPL, U.K. and visiting tutor with the University of Manchester) and R. Vijayakumar (Assistant Professor of Microbiology in the College of Science, Zulfi, Majmaah University in the Kingdom of Saudi Arabia).

This book is about cleanrooms and controlled environments in relation to the pharmaceutical and healthcare sectors. The book is applicable to both the sterile and non-sterile pharmaceutical sectors and its focus is upon cleanroom microbiology.

Modern approaches to contamination control place a greater emphasis upon environmental control than they do on upon monitoring. This requires an understand of risk assessment; hence risk management and contamination control strategies feature strongly in the text.

The book fills a much needed gap in the microbiology and contamination control spheres. While there are books on cleanrooms available, these focus almost entirely on the physical and rarely address microbiological risks. Similarly, there are various books on microbiology (even a few about pharmaceutical microbiology), yet these books rarely mention cleanrooms, or, where they do, give controlled environments limited coverage.

To the authors of Cleanroom Microbiology, these two domains, normally separated by different functions, are inseparable. This book is about cleanrooms and controlled environments in relation to the pharmaceutical and healthcare sectors and is applicable to both the sterile and non-sterile pharmaceutical sectors with its focus on cleanroom microbiology.

The book contains 16 chapters which cover a range of key topics. These include cleanroom standards, environmental monitoring, cleaning and disinfection, staff behaviours, understanding the microbiome of human skin, culture media, microbial identification and more.

The book has been published by PDA / DHI and details can be found here.

Posted by Tim Sandle

Sunday 18 January 2015

Researchers control adhesion of E. coli bacteria

Scientists have created a synthetic surface on which the adhesion of E. coli bacteria can be controlled. The layer, which is only approximately four nanometers thick, imitates the saccharide coating (glycocalyx) of cells onto which the bacteria adhere such as during an infection. This docking process can be switched on and off using light. This means that the scientists have now made an important step towards understanding the relationship between sugar (carbohydrates) and bacterial infections.

The bonding properties of the saccharide coating can now be switched using this method: if the researchers irradiate their system with light with a wavelength of 365 nanometres, considerably fewer pathogenic bacteria cells can adhere to the synthetic surface. The saccharide molecules turn away from the bacteria, in a sense, and can no longer be recognised. When switched on by 450 nanometre wavelength light waves, on the other hand, the structures reorientate such that the bacteria cells can dock on once again. In this way, E. coli adhesion can be controlled.

For details see:

Theresa Weber, Vijayanand Chandrasekaran, Insa Stamer, Mikkel B. Thygesen, Andreas Terfort, Thisbe K. Lindhorst. Switching of Bacterial Adhesion to a Glycosylated Surface by Reversible Reorientation of the Carbohydrate Ligand. Angewandte Chemie International Edition, 2014; DOI: 10.1002/anie.201409808

Posted by Tim Sandle

Saturday 17 January 2015

Influenza factsheet

Influenza is an acute viral infection of the respiratory tract. There are three types of influenza virus: A, B and C. Influenza A and influenza B are responsible for most clinical illness. Influenza is highly infectious with a usual incubation period of one to three days.

Public Health England has issued a factsheet about the viral disease. Here is an excerpt:

Changes in the principal surface antigens of influenza A – haemagglutinin and neuraminidase – make these viruses antigenically labile. Minor changes described as antigenic drift occur progressively from season to season. Antigenic shift occurs periodically, resulting in major changes and the emergence of a new subtype with a different haemagglutinin protein. Because immunity from the previous virus may not protect completely against the new subtype, the population may have little or no immunity, and this may therefore lead to widespread epidemics or even a pandemic.”

For further details see PHE.

Posted by Tim Sandle

Friday 16 January 2015

eBook: Biopharmaceutical Technical Resource Guide

Pharmaceutical Manufacturing has issued a new e-book of interest;

Few industries seem to have attained the luster and momentum that biopharma has over the last decade. This luster is attracting billions in recent capital investment, while producing dramatic gains in therapeutic success and better patient outcomes across several important disease categories.

In this edition of the Biopharmaceutical Technical Resources Guide, we’ve focused on delivering insight from some of the industry’s top players and opinion leaders as well as a few deep dives into technologies that support quality and attention to operations and process excellence — a winning strategy the whole industry should recognize.

For details see: PM

 Posted by Tim Sandle

Thursday 15 January 2015

Cataloguing 10 million human gut microbial genes

Over the last few years, research on bacteria in the digestive tract (gut microbiome) has confirmed the major role they play in our health. An international consortium has developed the most complete database of microbial genes ever created. The catalogue features nearly ten million genes and will constitute a reference for all research on gut bacteria.

Research on the gut microbiome (all of the bacteria in the digestive tract) has multiplied over the past several years, helped in great part by new sequencing technologies. The gut microbiome, which scientists have labelled a "new organ" that is composed of tens of trillions of bacteria -- ten times as many as the number of cells in the human body -- is directly linked to the immune system and brain. It is a major player in chronic illnesses such as obesity and Type 2 diabetes. However, research in the field depends on access to reference gene databases (or catalogues), which is particularly important when identifying the functions of microbial genes. Few and far between, existing catalogues were created using samples from a limited number of people and geographical origins.

INRA researchers, as part of the MetaHIT international consortium, put together a catalogue of microbial genes that regroups pre-existing catalogues (European, American and Chinese) and enhanced it with new sequences. Apart from being an unparalleled resource, the analyses done for the catalogue showed that it contains the broadest collection of microbial genes -- and by extension, their functions -- present in the global population. With ten million genes, this new catalogue presents the most impressive array of human intestinal bacteria in the world.

Most of the genes (around six million) are shared by just 1% of the population, making them quite rare. While there is substantial data today regarding the most common genes, future research will focus on determining the importance and role of these rare genes.

Thanks to this catalogue, the most clinically significant genes can be described, most notably those related to illnesses such as Type 2 diabetes, cirrhosis of the liver, cardiovascular diseases and some cancers. It will also provide a more complete picture of imbalances in the gut microbiome (dysbiosis), particularly those caused by medication.

For further details see:

An integrated catalog of reference genes in the human gut microbiome. Nature Biotechnology, 2014; 32 (8): 834 DOI: 10.1038/nbt.2942

Posted by Tim Sandle

Wednesday 14 January 2015

On the viral entry route

Insects can transmit viral diseases to humans. Therefore, understanding how insects cope with viral infection, and what immune mechanisms are triggered, can be important to stop diseases transmission. In a new study, researchers now show that the entry route of the virus changes how the insect host responds to it.

Using the fruit flies as a model of study, researchers have discovered an immune mechanism that is specifically effective when flies are infected through feeding.

When flies were fed with food containing virus they would require an immune mechanism that had been described only to be activated for infections caused by bacteria or fungi, the so called Toll pathway.

Knowing better the host's responses upon viral infection by different routes might also help to explain some biological phenomena observed in nature. For example, the survival of honeybees contaminated with virus seems to depend on the entry route of the virus. If contaminated through bites of mites, the honeybees die, whereas if they receive the virus from their progenitors, they can live.

For further details see:

Álvaro Gil Ferreira, Huw Naylor, Sara Santana Esteves, Inês Silva Pais, Nelson Eduardo Martins, Luis Teixeira. The Toll-Dorsal Pathway Is Required for Resistance to Viral Oral Infection in Drosophila. PLoS Pathogens, 2014; 10 (12): e1004507 DOI: 10.1371/journal.ppat.1004507

Posted by Tim Sandle

Tuesday 13 January 2015

New penicillin mechanism revealed

One of the oldest and most widely used antibiotics, penicillin, attacks enzymes that build the bacterial cell wall. Researchers have now shown that penicillin and its variants also set in motion a toxic malfunctioning of the cell's wall-building machinery, dooming the cell to a futile cycle of building and then immediately destroying that wall.

Penicillin and its variants also set in motion a toxic malfunctioning of the cell's wall-building machinery, which dooms the cell to a futile cycle of building and then immediately destroying that wall. This downstream death spiral depletes cells of the resources they need to survive.

There are two parts to the wall-assembly process: synthesizing new strands of linked sugars and then linking them into the expanding matrix. Beta-lactam drugs work by blocking the enzymes that build cross-links, weakening the wall. The wall cannot hold together, so the bacterial cell bursts and dies.

For further details see:

Hongbaek Cho, Tsuyoshi Uehara, Thomas G. Bernhardt. Beta-Lactam Antibiotics Induce a Lethal Malfunctioning of the Bacterial Cell Wall Synthesis Machinery. Cell, 2014; 159 (6): 1300 DOI: 10.1016/j.cell.2014.11.017

Posted by Tim Sandle

TB Alliance Announces Two New Members to its Board of Directors

TB Alliance Announces Two New Members to its Board of Directors:
Dr. Mario Raviglione of WHO and Shalini Sharp of Ultragenyx

TB Alliance, an international not-for-profit organization with the mission of developing improved tuberculosis (TB) treatments, announced the appointment of Mario Raviglione, MD, director of the Global TB Programme at the World Health Organization (WHO), and Shalini Sharp, the chief financial officer of Ultragenyx Pharmaceutical Inc., to its Board of Directors.

“Tuberculosis is a leading killer worldwide. The war against TB will only be won with new and better tools and treatments,” said Dr. Raviglione. “I’m deeply honored to join the Board of TB Alliance to contribute my knowledge and expertise, particularly as the organization prepares for late-stage testing and the potential introduction of new, innovative and game-changing TB treatments.”

“The TB Alliance’s product development partnership model is critical to the delivery of superior treatment options to patients in need,” said Ms. Sharp. “It is exciting to be working with an organization that has the potential to impact the course of this devastating disease and save many lives.”

Dr. Raviglione began working for the World Health Organization in 1991 and became director of the Global TB Programme in 2003. In his role, he is responsible for setting the policies and standards on global TB control, coordinating technical support, monitoring the global TB pandemic, and translating new evidence into policies and practice. Dr. Raviglione serves on a variety of scientific committees, has published extensively on the TB epidemic, and has won numerous awards for his work on TB control. Dr. Raviglione graduated from the University of Turin in Italy in 1980, and trained in internal medicine and infectious diseases in New York and Boston, where he was appointed an AIDS clinical research fellow at Beth Israel Hospital, Harvard Medical School.

Ms. Sharp has been the chief financial officer and senior vice president, finance, of Ultragenyx, a clinical-stage biotechnology company developing medicines to treat rare diseases, since 2012. She is a member of the Board of Directors of Agenus Inc. (formerly Antigenics Inc.), a publicly traded biotechnology firm, where she served as chief financial officer from 2006 to 2012. Prior to Agenus, Ms. Sharp held strategic planning and corporate finance roles at Elan Pharmaceuticals. She has also worked with McKinsey & Company and Goldman Sachs, specializing in pharmaceuticals and medical devices. Ms. Sharp holds both a BA and an MBA from Harvard University.

“We are pleased to welcome Dr. Raviglione and Ms. Sharp to the Board of the TB Alliance,” said Carlos Morel, MD, chairman of the board for TB Alliance and director of the Center for Technological Development in Health (CDTS) of FIOCRUZ, Brazil. “Their expertise will be invaluable as the organization launches an unprecedented number of trials and initiatives.”
Tuberculosis is a global pandemic, killing someone every 21 seconds—nearly 1.5 million in 2013 alone, according to the WHO. More than 95 percent of TB deaths occur in low- and middle-income countries, and TB is among the top five causes of death for women aged 15 to 44. Today's TB drug regimens take too long to cure the disease and, especially in the case of multidrug-resistant TB, are complicated to administer and can be toxic. New drug regimens are urgently needed to control TB and stop the spread of drug-resistant TB.

Posted by Tim Sandle

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