Thursday, 28 May 2015

ISO 14644 standards update

ISO 14644 is the international standard on cleanrooms. Once all parts are ratified it will consist of 15 parts, under the general title Cleanrooms and associated controlled environments. These are:

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 classification
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

Posted by Tim Sandle

Wednesday, 27 May 2015

ISO14644 Part 13

A new ISO 14644 cleanroom standard has reached the final draft stage. This is:
ISO 14644-13 - Cleanrooms and associated controlled environments -- Part 13: Cleaning of surfaces to achieve defined levels of cleanliness in terms of particle and chemical classifications.

This standard gives guidance on the selection of cleaning methods to achieve specified cleanliness levels. For the selection procedure, the aspects of surface description, cleanliness specifications, types of contamination, cleaning techniques, material compatibility, and assessment methodology are taken into consideration.

The standard provides guidance on the assessment of cleaning methods for achieving the required surface cleanliness by particle concentration (SCP) and surface cleanliness by chemical concentration (SCC) classes and which techniques should be considered to achieve these specified levels.

Copies of the draft are available from national standard organizations.

Posted by Tim Sandle

Tuesday, 26 May 2015

Best Practices in Pharmaceutical Microbiology - India

Pharmig are set to run the first international pharmaceutical microbiology course in India. The events take place:
  • Tuesday 28th July 2015– Biocon – Bangalore
  • Thursday 30th July 2015– Orchid Hotel – Mumbai
The course features: Scott Sutton, Rachel Blount, Julie Roberts, Ken Muhvich

The topics covered include:
  • Regulatory Hot Topics and Warning Letters
  • Data Integrity
  • QC Microbiology and the Compendia
  • Validating Disinfectant Efficacy & Test Methods
  • Objectionable Organisms Within & Beyond Regulations for Steriles   & Non Steriles
  • Implementing a Disinfectant Programme: A Practical Approach
  • Developing a Meaningful Environmental Monitoring Programme
  • Microbiology Laboratory Investigations - Errors, Preparation and Success
Posted by Tim Sandle

Monday, 25 May 2015

Laboratory Techniques with Applicability in Medical Practice

A new book of interest has been published, edited by Tamás Kőszegi and Antonella Chesca. The book presents medical information containing clinical and experimental data referring to state-of-the-art methods used in laboratory medicine for diagnostic and follow-up purposes.

The book includes chapters on microbiological screening methods, bacterial endotoxin, genome sequencing, sepsis investigations, orosomcoids, tumour cells, electrophoreses, and melanocytic nevi.

The book includes a chapter by Tim Sandle on endotoxin testing and the LAL assay:

Sandle, T. (2015) Bacterial Endotoxin Testing using the Limulus Amebocyte Lysate Assay. In Kőszegi, T. and Chesca, A. (Eds.) Laboratory Techniques with Applicability in Medical Practice, Lambert Academic Publishing, pp19-32

For further details, please contact Tim Sandle

Posted by Tim Sandle

Sunday, 24 May 2015

Salmonella survives the macrophage's acid attack

Macrophages destroy bacteria by engulfing them in intracellular compartments, which they then acidify to kill or neutralize the bacteria. However, some pathogenic bacteria, such as Salmonella enterica, have evolved to exist and even grow while within these acidified compartments. Yet, how Salmonella responds to the acidic environment and how that environment affects the virulence of this pathogen are unclear. New research reveals that Salmonella fights acid with acid, by lowering the pH of its own interior in response to the acidification of the Salmonella-containing compartment by the macrophage, and by using that low pH as a signal to turn on genes needed to establish an infection.

To investigate the effect of the acidic environment on Salmonella, the authors used a biosensor, called an I-switch, which allowed for measurements of pH within a single cell. Using the I-switch, the authors found that the Salmonella cytoplasm acidifies rapidly after being engulfed and exposed to the acidic environment of the macrophage interior. Interestingly, they found that the Salmonella actively, as opposed to passively, acidify their cytoplasm.

Researchers found that bacteria can survive a cytoplasmic pH of 5.6 and that they even use this to signal expression of virulence genes. The research shows that low pH activates an intracellular signalling cascade, which induces the formation of a nanomachine called the type III secretion system. This nanomachine is composed of a needle complex used to inject bacterial virulence proteins into the host cell.

For further details, see Phys.Org.

Posted by Tim Sandle

Saturday, 23 May 2015

Protecting Antibiotics

Only one quarter of countries have an action plan to address antibiotic resistance. This according to a new survey by the World Health Organization (WHO).
The headline from the WHO report is that around the world the sale of antibiotics over-the-counter is “widespread.” Furthermore, it is very rare that nations track resistance to the drugs or put in place measures to slowdown their over-use.
With the report, WHO surveyed 133 national governments to assess their plans for the growing threat of antibiotic resistance. Of these, only thirty-four reported that they had a detailed national plan in place. This number included four of 26 responding countries in the Western Pacific, five of 11 countries in southeast Asia, 40 percent of European respondents. Interestingly, none in the eastern Mediterranean region nations and only three of 26 countries in the Americas had any measures. In Africa, the data were incomplete, but responses from eight countries indicated a growing problem of antibiotic resistance.

For more details, see TLN.

Posted by Tim Sandle

Friday, 22 May 2015

Chagas Disease Vaccine Promise

Chagas disease (or American trypanosomiasis) causes fever, fatigue, body aches, headache, rash, loss of appetite, diarrhoea, and vomiting. As well as growing in cases in the U.S. (particularly in Texas), it is a major cause of heart disease and gastrointestinal dysfunction in widespread areas throughout Latin America.
Chagas disease is caused by the tiny worm-like parasite Trypanosoma cruzi, which is carried by so-termed Kissing Bugs (the insects tend to leave bites around the mouth, specially while people are sleeping at night). Additionally, there is some evidence that bed bugs can also carry the parasite.

Currently there is no vaccine available. Treatment tends to be with anti-parasitic drugs (for those fortunate enough to be able to afford them).

In a new study, researchers report on success in developing the vaccine in mice infected with the disease causing parasite. Research has been led by scientists based at the University of Texas Medical Branch in Galveston, USA.

For more details, see TLN.

Posted by Tim Sandle

Thursday, 21 May 2015

Considerations of melanocytic nevi in children

The incidence of melanocytic nevi is increasing, and nevocytic nevi are commonly encountered in childhood. Therefore, the study of these tumors, particularly those that are likely to become malignant, represents an important issue.

With this regard, A. Chesca, M.C. Luculescu and Tim Sandle have written a paper. The abstract is:

“This paper presents clinical, statistic and structural data on the melanocytic nevi in children, and of adjacent areas when the nevi were surgically excised. The study was made possible by collaboration with the specialty medical stuff of Children Clinic Hospital Braşov. The analysis of the melanocytic nevi was retrospective in relation to 30 patients. These cases were selected from the cases investigated in the first half of 2013. The pathological anatomy diagnosis was conducted following a biopsy of the fragments of melanocytic nevi excised by surgery. This took place within the plastic surgery department of the Children Clinic Hospital Brasov.”

The reference is:

Chesca, A., Luculescu, M.C. and Sandle, T. (2014) Considerations of melanocytic nevi in children, Annals of R.S.C.B., Vol. XIX, Issue 2, 2015, pp. 19 – 22 (doi: 10.ANN/RSCB-2015-0001:RSCB)

The article can be accessed here

Posted by Tim Sandle

Wednesday, 20 May 2015

African Centres for Disease Control and Prevention (CDC)

The African Centres for Disease Control and Prevention (CDC) is set to open this year, Time has reported.

The news has been welcomed by the U.S. equivalent body. “The West African Ebola epidemic reaffirmed the need for a public health institute to support African ministries of health and other health agencies in their efforts to prevent, detect, and respond to any disease outbreak,” US CDC Director Tom Frieden said in a statement.

Posted by Tim Sandle

Tuesday, 19 May 2015

Ebola in West Africa - the way forwards

A special webcast on Ebola is available to view online.

More than a year into the Ebola outbreak in West Africa, Doctors Without Borders/Médecins Sans Frontières (MSF) continues to respond  in the affected countries. Although the number of recorded cases has decreased, much work remains to be done — the outbreak has ravaged health systems, forcing many medical facilities to close and killing hundreds of health personnel.  While non-Ebola health concerns must be addressed, from maternal health care to vaccinations, epidemiological surveillance and community awareness of Ebola must be improved to ensure that the outbreak is stopped.

The unprecedented epidemic has also exposed the weaknesses of the international response and pushed MSF to extremes. Since helping to identify the epidemic in March 2014, MSF staff members have played a key role in the response, from health promotion to contact tracing and patient care.
A panel of MSF staff members and recently returned aid workers will discuss current challenges and lessons learned in the Ebola epidemic.

A recording of this webcast, which aired May 14, is now available online at MSF.


Sophie Delaunay is the executive director of MSF-USA. In response to the Ebola outbreak, she directed MSF’s advocacy efforts with the U.S. government, and oversaw the management of U.S. field workers who were departing and returning from Ebola projects. Most recently, she has been leading MSF’s Ebola initiative, which explores what role MSF can have in future Ebola-related research and development projects, while ensuring that any initiative would be driven by patient needs and would respect the ownership rights of West African governments. During the outbreak, she visited Liberia and Senegal.

Ella Watson-Stryker began working for MSF in March 2014 as part of MSF’s initial response to the Ebola outbreak in Guinea. She returned for subsequent missions as an MSF health promotion manager in Sierra Leone and Liberia, and was recently featured in Time magazine with other health care providers and aid workers in West Africa who were collectively named Person of the Year for 2014.

Dr. Gillian Burkhardt is an obstetrician and gynecologist, and recently returned from Sierra Leone, where she worked in an Ebola treatment center that was transitioning into a center that will specifically treat pregnant Ebola patients. Her first assignment for MSF was at the beginning of 2014 in Sierra Leone prior to the Ebola outbreak, and in the same year, she also worked for MSF in the Ivory Coast.
Dr. Craig Spencer worked as a physician in an Ebola treatment center in Guinea in the fall of 2014. 
Following a successful recovery from Ebola, he completed a second assignment with MSF in Guinea this spring, working as an epidemiologist.

Posted by Tim Sandle

Monday, 18 May 2015

European Pharmacopeia updates

Some updates of interest in relation to the European Pharmacopeia:

European Pharmacopoeia 8th Edition - Supplement 8.5 (implementation date 1st July 2015)

General Chapters

2.2.32  Loss on drying                                                                       

Clarification of the way instrument qualification is to be performed.

2.5.12  Water: semi-micro determination                                           

Clarification of the way instrument qualification is to be performed.

2.7.18  Assay of human coagulation factor II                                   

EDTA has been removed from the dilution buffer because its presence has been reported to inhibit the ecarin-dependent coagulation factor II activation and cause a decrease in the sensitivity and precision of the assay.

Posted by Tim Sandle

Sunday, 17 May 2015

Thermonuclease Test explained

This test is also known as the heat stable nuclease test. It is a 4hr test based on the production of a heat stable DNase (thermonuclease) by Staphylococcus aureus.

It is also used for determining and confirming the presence of S. aureus subsp aureus DNase from that produced by S. epidermidis or other micrococci. It is of particular use in determining the presence of S. aureus in positive blood culture bottles.

Unlike other staphylococci, most strains of S. aureus and Staphylococcus intermedius produce thermonuclease, a heat stable DNase.

Subspecies of Staphylococcus schleiferi are DNase positive and produce heat stable nucleases. The thermonuclease test detects the presence of this DNase.

The organism is heated to destroy heat labile thermonucleases. It is then inoculated on medium containing DNA and toluidine blue. The DNA is broken down by heat stable nucleases resulting in the toluidine blue changing to red or pink.

In relation to the thermonuclease test, Public Health England has issued a technical report, including safety information. The report can be accessed here.

Posted by Tim Sandle

Saturday, 16 May 2015

Cord Blood Market: The 4 key metrics

For those managing a cord blood bank or investing in the cord blood market, the market can initially appear technical and confusing. However, there are four key metrics to assess. These metrics are:

1.Hematopoietic Stem Cells Transplant Rates  – Determine the number of hematopoietic stem cell transplants occurring per year and quantify year-over-year changes.

2.Rates of Cord Blood Utilization – Determine the percentage of hematopoietic stem cell transplants (from all sources) contributed by cord blood stem cells.

3.Frequency of Cord Blood Stem Cell Transplant, Relative to Other Alternatives – Identify the frequency at which cord blood stem cell transplants are performed, relative to the other common stem cell transplant alternatives.

4.Rates for Cord Blood Banking Awareness – Quantify existing levels of market participating and the potential for future market expansion.

To help with this, Total Biopharm have produced a useful infographic that can be accessed here.

Posted by Tim Sandle

Friday, 15 May 2015

New approach to help prevent meningococcal outbreaks

Nasal drops of harmless bacteria can inhibit a related bug that sometimes causes meningococcal disease, according to new findings published online in Clinical Infectious Diseases. The study--conducted among college students, a group at higher risk for this often serious illness--suggests a new approach that could help suppress outbreaks of the disease, if supported by future research.

Meningococcal disease is caused by Neisseria meningitidis, which can infect the lining of the brain and the spinal cord, causing meningitis. Strains of the bacteria can also cause serious bloodstream infections. But N. meningitidis can also live silently in a person's nose and throat, without illness. These "colonized" carriers can spread the pathogen to others through close contact.

In the study, researchers placed drops containing low doses of Neisseria lactamica, a related but harmless bacterial strain, into the noses of 149 healthy university students in the United Kingdom. A control group of 161 students received drops of saline instead. Nose swabs were taken at regular intervals over six months and tested for both types of bacteria.#

For further details, see Medical News

Posted by Tim Sandle

TB Alliance Launches “Nix-TB” Clinical Trial to Test New XDR-TB Treatment

First clinical trial to test a new all-oral regimen for extensively drug-resistant TB; Nix-TB is a critical step in the development of a universal treatment for all types of TB

TB Alliance and its partners announced the start of a clinical trial of a new regimen to treat extensively drug-resistant tuberculosis (XDR-TB.) It is the first study to test an all-oral drug regimen, comprised of drugs with minimal pre-existing resistance, that has the potential to shorten, simplify, and improve treatment for XDR-TB.

“XDR-TB is an absolute devastation to patients, their families, and communities. The study is the first to test a novel and potentially transformative regimen for XDR-TB, which could be a valuable tool as we battle this problem on the front lines in South Africa and around the world,” said Francesca Conradie, MD, the Principal Investigator of the Nix-TB (New Investigational Drugs for XDR-TB) trial and Clinical Advisor at Sizwe Hospital, in Johannesburg, South Africa. “The strategic emphasis by our National Department of Health on clinical research for drug-resistant TB coupled with a rigorous regulatory framework has enabled this trial to be conducted in South Africa.”

XDR-TB is a strain of tuberculosis, airborne and infectious, that has resulted from progressive antibiotic resistance and is resistant to four commonly used anti-TB drugs. XDR-TB has been reported in 100 countries. It is complicated and expensive to treat and results in high rates of death. Today, there are no regulatory-approved XDR-TB treatments.

Currently, healthcare providers treat XDR-TB by individualizing treatment regimens, frequently using antibiotics not normally used for TB as well as toxic medicines not meant for the long treatment duration that TB requires. People with XDR-TB can be on treatment for two years or longer, with thousands of pills and injections, extensive side effects, and little success.
 In a recent review of the experience in South Africa, after two years of treatment only a fraction of people—16 percent—with XDR-TB were cured.

Resistance to available antibiotics has plagued efforts to combat the TB pandemic, creating distinct drug-resistant strains of the bacteria such as multi-drug resistant TB (MDR-TB) and XDR-TB and rendering current treatments inadequate. However, the three drugs that comprise the treatment being tested in Nix-TB have novel mechanisms of action. The three-drug regimen consists of bedaquiline (B), which received conditional regulatory approval in several high-TB disease burden countries; the novel antibacterial drug compound pretomanid (Pa), which is being tested in multiple clinical trials; and linezolid, an oxazolidinone, which has been used off-label to treat TB.

If the regimen tested in Nix-TB is successful and safe in XDR-TB, that will pave the way for expanding the study, testing its potential for use in people with MDR-TB and then potentially in people with drug-sensitive TB. Having a regimen that would be usable in such a broad range of TB patients could significantly improve TB control efforts globally.

“We are testing a promising treatment for XDR-TB today, but the longer-term potential of such a regimen is even greater. We now see the possibility of a single TB regimen that can treat virtually all patients with active TB with a relatively simple and affordable regimen,” said Mel Spigelman, MD, President and CEO of TB Alliance. “The launch of Nix-TB is a critical step to achieve the vision of a truly short-course, simple, affordable and well-tolerated universal treatment regimen.”

Nix-TB is a partnership between TB Alliance, a not-for-profit organization with the mission of developing improved TB treatments and the sponsor of the trial; Janssen Pharmaceutica NV (Janssen), the originator company that in 2009 granted a royalty-free license to the TB Alliance for the development and commercialization of bedaquiline in the field of drug-susceptible TB; and the sites in South Africa where the study is and is expected to be conducted (Sizwe Hospital, TASK at Brooklyn Chest Hospital, and THINK at Doris Goodwin Hospital).

The cost for the initial phase of Nix-TB is covered by a group of long standing TB Alliance donors. TB Alliance is starting to bring together additional funding to expand the study and the number of sites.
“The availability of new TB drugs offers the unprecedented opportunity to improve treatment for people with TB. However, the existence of individual new drugs is not enough,” said TB Alliance’s Spigelman. “TB must be treated in multi-drug combinations or regimens to enhance efficacy and prevent the development of resistance. Therefore, the Nix-TB trial fills a critical gap and capitalizes on the availability of novel drugs by studying them together in the most vulnerable TB population, those with XDR-TB, and in such a way that provides clear understanding of how to use the treatments to maximize their impact on the epidemic.”

Posted by Tim Sandle