Friday, 24 October 2014

Antibacterial resistance a cause for major concern

Professor Stuart Elborn, an international authority on respiratory medicine, said that more funding and further research are required into antibiotic resistance in order to improve patient outcomes for people with Cystic Fibrosis.

In his paper, Infections in chronic lung diseases 2, which was recently published in The Lancet, Professor Elborn reviews current research into infections in chronic lung diseases. Professor Elborn and his colleagues state that while not all resistance found in bacteria is caused by antibiotics, the increasing resistance to antibiotics is proving a major problem in treating people with Cystic Fibrosis.

Professor Elborn stated: “We need more research into how to improve cystic fibrosis patient outcomes while reducing antibiotic resistance. We need to look at the use of compounds that may work against bacteria in a way that helps our current antibiotics to be more effective. Such compounds are readily available for treatment of other conditions. At Queen's we are leading the way and are working on developing some of these compounds.”

For further details see:

Laura J Sherrard, Michael M Tunney, J Stuart Elborn. Antimicrobial resistance in the respiratory microbiota of people with cystic fibrosis. The Lancet, 2014; 384 (9944): 703 DOI: 10.1016/S0140-6736(14)61137-5

Posted by Tim Sandle

Thursday, 23 October 2014

PCR for food microbiology

Real-time PCR assays for food microbiology have been developed into commercial products whereby some or all of the steps can be automated to minimise the number of operations involved and reduce the risk of contamination. Automation requires a higher capital investment and so will depend on the sample throughput required. The reaction usually takes place inside a computer-controlled combined thermocycler/fluorescence detection instrument and uses pre-prepared reagents. For foodborne pathogen detection tests, the entire process can be completed within 20-30 hours with sufficient sensitivity to detect a single cell in a 25g sample.

The main advantage for PCR-based methods is in shorter detection times, but the high degree of automation built into PCR systems also allows relatively unskilled staff to run them without extensive training. The high specificity of PCR can also mean fewer repeat tests. The principal disadvantage is currently cost, both in terms of capital outlay and consumables. While larger laboratories can benefit from reduced labour costs, economies of scale and rapid results, the benefits for smaller labs may be less clear.

A number of commercial PCR systems are currently offered for food pathogen detection. One of the first into the marketplace was the Bax® system from Dupont Qualicon and this has been joined by TaqMan® and MicroSEQ® food pathogen detection kits from Life Technologies, iQ Check real-time PCR kits from Bio-Rad, foodproof® real-time PCR detection kits distributed by Merck, and several others.

One of the main reasons for the comparatively high capital cost of PCR-based detection systems is the need for thermocycling during the amplification step. Instruments must be capable of very accurate and precise temperature control throughout the cycle. In addition to the capital investment for the instrumentation, the chemistry is also expensive as it uses fluorescent probes. Novel developments in DNA synthesis have demonstrated alternative solutions for DNA amplification under isothermal conditions without the need for a thermocycler. There are several types of nucleic acid amplification technologies. Amongst the isothermal DNA amplification technologies that have been developed, loop-mediated isothermal amplification (LAMP) has been used widely to detect microorganisms and is a promising and suitable technology for the rapid detection of pathogen in the field. LAMP uses multiple primers and a bacterial polymerase, Bst polymerase, derived from Bacillus stereothermophilus to amplify DNA rapidly at a constant 63oC. This does away with the need for a thermocycler component in the instrument and can reduce the cost by up to two thirds.

A commercial isothermal amplification system has already been developed for food pathogen detection. The 3M™ Molecular Detection System uses a unique bioluminescence method to detect the amplification of DNA sequences and is designed to be simple to use. The amplification and detection processes are completed within 75 minutes with real-time positive results available as early as 15 minutes. An overnight single enrichment step is still required at present. Test kits for E. coli O157 including H7, Salmonella and Listeria spp. detection in food and environmental samples are currently available.

Portable PCR-based instruments have also been developed recently. For example, Idaho Technology markets the ‘Ruggedized Advanced Pathogen Identification Device’ or R.A.P.I.D.® System, which uses an air thermocycling process and a fluorimetric detection system to detect Salmonella, Listeria, E. coli O157 and Campylobacter in food samples.
Posted by Rapid Microbiology

Wednesday, 22 October 2014

Scientists Discover an On/Off Switch for Aging Cells

Scientists at the Salk Institute have discovered an on-and-off “switch” in cells that may hold the key to healthy aging. This switch points to a way to encourage healthy cells to keep dividing and generating, for example, new lung or liver tissue, even in old age.

In our bodies, newly divided cells constantly replenish lungs, skin, liver and other organs. However, most human cells cannot divide indefinitely–with each division, a cellular timekeeper at the ends of chromosomes shortens. When this timekeeper, called a telomere, becomes too short, cells can no longer divide, causing organs and tissues to degenerate, as often happens in old age. But there is a way around this countdown: some cells produce an enzyme called telomerase, which rebuilds telomeres and allows cells to divide indefinitely.

In a new study published September 19th in the journal Genes and Development, scientists at the Salk Institute have discovered that telomerase, even when present, can be turned off.

Posted by Tim Sandle

Tuesday, 21 October 2014

Analytical & Bioanalytical Testing

Effective analytical instruments, methods, and testing services are vital to all phases of drug development. In this special e-book experts explain methods of identifying viral contaminants, assays for comparability testing of biosimilars, method transfer, data management, and assessing protein purification.

This is the basis of a new, free e-book from BioPharm

The book contains the following articles:
  • Viral Contamination: The Challenge of Finding the Unknown
  • Testing the Quality, Safety, and Efficacy of Insulin Biosimilars
  • Keys to Successful Method Transfer
  • Standardizing Data Management
  • Using the Purity Quotient Difference to Assess Protein Purification

For details, see BioPharm

Posted by Tim Sandle

Cleanroom Clothing Practices in the Healthcare and Pharmaceutical Sector

Pharmig presents:
Cleanroom Clothing Practices in the Healthcare and Pharmaceutical Sector
Tuesday 28th October 2014
Best Western Plus Windmill Village Hotel, Golf & Spa, Coventry

09.00 – 09.30                       Registration with tea/coffee
09.30 – 09.45                       Chairs introduction
                Dr. Tim Sandle – BPL and Pharmig Committee Member

09.45 – 10.30                       Microbiological regulatory aspects of garments and gloves
                                                Speaker: Greg Cochran – Micronclean

10.30 – 11.15                       Microbiological risk to cleanrooms and disinfection
-          Cleanrooms
-          Contamination and contamination sources
-          Selecting detergents and disinfectants
                                                Speaker: Dr. Tim Sandle – BPL & Pharmig Committee Member

11.15 – 11.45                       Morning break with tea/coffee

11.45 – 12.30                       Cleanroom garments for effective ongoing contamination control
                                                -      Personnel contamination and Regulatory Authority requirements
-          Requirements for effective cleanroom garments and assessment for garment lifetime
-          Gowning procedures and validation
-          Garments and cleanroom energy considerations
                                                Speaker: Tim Eaton – AstraZeneca

12.30 – 13.30                       Lunch

13.30 – 14.15                       Gloves and standards
-          What are the ideal properties of gloves for aseptic properties
-          Weaknesses of gloves
-          Use of gloves in isolators
-          Comparisons between American and European standards
-          Chemical and microbiological testing
-          Cytotoxic testing
-          Practical approach to gloves and standards
                Speaker: Mark Oldcorne – Wrexham Maelor Hospital

14.15 – 15.00                       Validation and Application of Hand Hygiene Products
                Speaker: Laura Guardi – ECOLAB

15.00 – 15.20                       Afternoon break with tea/coffee

15.20 – 16.00                       Q&A with the Expert Speaker Panel & Conducting a Cleanroom Clothing Survey
This session will be dedicated to delegates where they can ask speakers additional                                                                                questions relating to Cleanroom Clothing issues.                                          
                                                We also ask that within this session you kindly take the time to complete a questionnaire                                                    on Cleanroom Clothing –the aim of which is to look to produce a ‘Best Practice and Issues                                                   around Cleanroom Clothing in the Healthcare and Pharmaceutical Sectors’ as an article                                                                     and/ or publication

16.00 – 16.15                       Chair’s closing remarks

For details contact Pharmig

Posted by Tim Sandle

Monday, 20 October 2014

Exploring pulmonary pathology

The test of the pulmonary function represents an auxiliary method for the specialty medical examination the patient to determine if there are signs and symptoms relevant to a pulmonary pathology. This is the basis of a new article by Antonella Chesca and Tim Sandle.

The abstract runs:

“This study evaluates the respiratory parameters associated with pulmonary diseases. The study was centred on a comparative evaluation of the pulmonary function during the same two periods of two different years: January - February 2013 and January – February 2014. The data was analysed and comparison was undertaken between the severity of the pulmonary diseases symptoms requiring spirometries, during the cold season in two succeeding years. The study applied to symptomatic patients suffering from COPD or asthma who were referred to the Specialty Outpatient Department of the Pneumophthisiology Hospital of Brasov. To determine the diagnosis and to apply the appropriate medication, the study examined the function tests by spirometry for each of the patients in the study group. As applicable, spirometry was carried out by tests using salbutamol, according to the orders of the specialty doctor. The analysis of the study data enabled the creation of the profile of the symptomatic patients.”

The reference is:

Chesca, A. and Sandle, T. (2014) Data on the examination of functions in pulmonary disease, Acta Medica Transilvanica, 2 (3): 208 – 210

For a copy, please contact Tim Sandle

Posted by Tim Sandle

Antibiotic resistance continues to rise

“Antibiotic resistance continues to rise,” BBC News reports as, despite warnings, the number of antibiotic prescriptions in the UK continues to soar, as do new cases of resistant bacteria.

The news follows the publication of a new report by Public Health England on the English surveillance programme for antimicrobial utilisation and resistance (ESPAUR), which reports the change in antibiotic prescribing and resistance over recent years.

The report highlights a number of key findings, including a year on year increase in antibiotic prescribing in England, with the majority of antibiotic prescribing taking place in general practice. There also seems to be variation across the UK, with areas with higher antibiotic prescribing also having higher rates of resistant infections.

Public Health England hopes this report will enable general practices and hospitals to compare their data with regional and national trends. They could then see if their rates are higher than other areas, and investigate why and if they can reduce these. This will also provide a baseline measure from which to track changes in both prescribing and resistance in England.

Posted by Tim Sandle

Sunday, 19 October 2014

Entire genome of a Klebsiella pneumoniae strain sequenced

Microbiologists, for the first time, recently sequenced the entire genome of a Klebsiella pneumoniae strain, encoding New Delhi Metallo-beta-lactamase (NDM-1).

Assembling an entire genome is like putting together a puzzle. Klebsiella pneumoniae turned out to have one large chromosome and four plasmids, small DNA molecules physically separate from and able to replicate independently of the bacterial cell's chromosomal DNA. Plasmids often carry antibiotic resistant genes and other defense mechanisms.

The researchers discovered their Klebsiella pneumoniae bacteria encoded 34 separate enzymes of antibiotic resistance, as well as efflux pumps that move compounds out of cells, and mutations in chromosomal genes that are expected to confer resistance. They also identified several mechanisms that allow cells to mobilize resistance genes, both within a single cell and between cells.

For further details, see:

Corey M. Hudson, Zachary W. Bent, Robert J. Meagher, Kelly P. Williams. Resistance Determinants and Mobile Genetic Elements of an NDM-1-Encoding Klebsiella pneumoniae Strain. PLoS ONE, 2014; 9 (6): e99209 DOI: 10.1371/journal.pone.0099209

Posted by Tim Sandle

Saturday, 18 October 2014

Fostering Cooperation and Strengthening Medical Product Regulatory Systems in the Americas

From the FDA:

Office of International Programs (OIP) has announced the availability of grant funds for the support of a single source cooperative agreement to the Pan American Health Organization (PAHO) for fostering cooperation and strengthening medical product regulatory systems in the Americas. The goal of the cooperative agreement is to build upon existing cooperation between OIP/FDA and PAHO to foster regulatory collaboration and strengthen regulatory capacity throughout the Americas.

See: FDA

Posted by Tim Sandle

Friday, 17 October 2014

FDA Issues New Guidance: Why Your ANDA was Refused?

The FDA has issued two guidance documents intended to assist applicants preparing to submit abbreviated new drug applications (ANDA), clarifying the deficiences that may cause the FDA to refuse-to-receive (RTR) the submissions.

The two guidances included a final guidance on ANDA RTR standards and a draft guidance on ANDA RTR for lack of proper justification of impurity limits.

According to the FDA, between 2009 and 2012, the Office of Generic Drugs refused to receive 497 ANDAs, primarily because the submissions contained serious deficiences.

FDA’s ANDA RTR standards guidance finalizes a draft version of the guidance released in October 2013. In the ANDA RTR impurity limits draft guidance, the FDA describes serious deficiencies in impurity information that could cause FDA to refuse-to-review an ANDA.

According to the draft guidance, typical deficiencies leading to a refuse-to-receive decision include:

(1) failing to provide adequate justification for proposed limits in drug substances and drug products for specified identified impurities that are above qualification thresholds
(2) failing to provide adequate justification for proposed limits for specified identification thresholds
(3) proposing limits for unspecified impurities (e.g., any unknown impurity) that are above identification thresholds. unidentified impurities that are above identification thresholds.

Read the draft guidance here and the final guidance here.

Posted by Tim Sandle

Thursday, 16 October 2014

Using biofilms for society’s advantage

Biofilms are communities of bacteria ensconced in a slimy, but extremely tough, matrix of extracellular material composed of sugars, proteins, genetic material and more. During biofilm formation individual bacteria pump out proteins that self-assemble outside the cell -- creating tangled networks of fibers that essentially glue the cells together into communities that keep the bacteria safer than they would be on their own.

Normally considered a bad thing, biofilms also self-assemble and self-heal. In looking at this, researchers have genetically fused a protein with a particular desired function -- for example, one known to adhere to steel -- onto a small protein called CsgA that is already produced by E. coli bacteria. The appended domain then went along for the ride through the natural process by which CsgA gets secreted outside the cell, where it self-assembled into supertough proteins called amyloid nanofibers. These amyloid proteins retained the functionality of the added protein -- ensuring in this case that the biofilm adhered to steel.

This could be useful in materials science. It could be possible to produce a raw material as a building block, they orchestrate the assembly of those blocks into higher order structures and maintain that structure over time.

For further details see: Wyss Institute for Biologically Inspired Engineering atHarvard.

Posted by Tim Sandle

Wednesday, 15 October 2014

Spherical influenza virion

A spherical influenza virion is an orderly hodgepodge comprised of hundreds of proteins that originate from both the virus and its host. A new mass spectrometry analysis, published in Nature Communications, has yielded the most complete picture to date of the identities, arrangements, and ratios of proteins in influenza virions.

To sort out the virion components, researchers used sensitive mass spectrometry. Comparing the intensities of ion spectra among protein fragments allowed them to quantify the relative abundance of each protein.

For further details, see The Scientist.

Posted by Tim Sandle

Tuesday, 14 October 2014

ISO 14644 – standard list

ISO 14644 is a multi-part cleanroom standard.

ISO 14644 consists of the following parts, under the general title “Cleanrooms and associated controlled environments”:
  • Part 1: Classification of air cleanliness by particle concentration
  • Part 2: Monitoring to provide evidence of cleanroom performance related to air cleanliness by particle concentration
  • Part 3: Test methods
  • Part 4: Design, construction and start-up
  • Part 5: Operations
  • Part 7: Separative devices (clean air hoods, gloveboxes, isolators and mini-environments)
  • Part 8: Classification of air cleanliness by chemical concentration (ACC)
  • Part 9: Classification of surface cleanliness by particle concentration
  • Part 10: Classification of surface cleanliness by chemical concentration
  • Part 12: Classification of air cleanliness by nanoscale particle concentration
  • Part 13: Cleaning of surfaces to achieve defined levels of cleanliness in terms of particle and chemical classifications
  • Part 14: Assessment of suitability for use of equipment by airborne particle concentration
  • Part 15: Assessment of suitability for use of equipment and materials by airborne chemical and surface chemical concentration
The reader should note that there is no 'Part 11' currently in development.

The standards use the following common codes:

APC – Airborne Particulate Cleanliness
SPC – Surface Particulate Cleanliness
AMC – Airborne (Chemical) Molecular Cleanliness
SCC – Surface Chemical Cleanliness
AVC – Airborne Viable Cleanliness
SVC – Surface Viable Cleanliness

Copies are available from national standards agencies.

Posted by Tim Sandle

Monday, 13 October 2014

Sanitation of Pharmaceutical Facilities

Maintaining environmental control including microbiological contamination in a pharmaceutical manufacturing environment is primarily dependent on the facility sanitization program. Sanitization considerations are specific for facility rooms, equipment, and personnel. Sanitization comprises cleaning and disinfection. Cleaning is necessary prior to the application of disinfectant to enable sufficient contact time of the disinfecting agent with the surface.

In a new paper, Tim Sandle provides an introduction to the sanitization and bio-decontamination of pharmaceutical manufacturing facilities. This topic is especially relevant for manufacturing of sterile products.

The reference is:

Sandle, T.  (2014) Sanitation of Pharmaceutical Facilities, Journal of GXP Compliance, 18 (3): 6-10

For details see: IVT

Posted by Tim Sandle

Sunday, 12 October 2014

Photosynthetically productive light distributed to symbiotic microalgae

Iridescent cells in the mantle tissue of giant clams spread light of a wavelength that drives photosynthesis to microalgae that provide nutrition for the animals, the University of Pennsylvania’s Alison Sweeney and colleagues reported in Journal of the Royal Society Interface.

In their paper, the researchers likened the symbiotic system to an electric transformer, “which changes energy flux per area in a system while conserving total energy.” Given this parallel, the authors proposed that the clam system might inspire the development of more efficient and resilient photovoltaic materials.

These so-called iridocytes not only distribute photosynthetically productive light to the algae, they also reflect nonproductive light, the researchers showed. “At incident light levels found on shallow coral reefs, this arrangement may allow algae within the clam system to both efficiently use all incident solar energy and avoid the photodamage and efficiency losses,” the researchers wrote in their paper.

Source: The Scientist