Monday, 31 March 2014

Novel Methods to Address Antimicrobial Resistance


Humans face the very real risk of a future without antibiotics. The implications of this are that life expectancy could fall due to people dying from diseases that are readily treatable today. One concern for the future is the re- emergence of diseases that are ‘officially’ extinct, such as tuberculosis (a disease caused by Mycobacterium tuberculosis). The other concern is that without effective antibiotics, human society will no longer be able to conduct the types of medical procedures that can lead to immunosuppression. Such therapies include those for cancer treatments or to address autoimmune disorders.

This is the basis of a review article written by Tim Sandle and published in the journal SOJ Microbiology & Infectious Diseases. The reference is:



Sandle T (2014) Novel Methods to Address Antimicrobial Resistance. SOJ Microbiol Infect Dis 2(1): 2

To view a copy, please contact Tim Sandle

Posted by Tim Sandle

Sunday, 30 March 2014

Prickly protein helps stop Staphylococcus


A genetic mechanism that controls the production of a large spike-like protein on the surface of Staphylococcus aureus bacteria alters the ability of the bacteria to form clumps and to cause disease, according to a new study.

Under normal conditions, Staphylococcus bacteria interact with proteins in human blood to form aggregates, or clumps. This clumping behavior has been associated with pathogenesis -- the ability of bacteria to cause disease. However, the mechanisms that control clumping are not well understood.

In the process of investigating how staph bacteria regulate cell-to-cell interactions, researchers at the UI Carver College of Medicine discovered a mutant strain of staph that does not clump at all in the presence of blood proteins.

Further investigation revealed that the clumping defect is due to disruption of a genetic signaling mechanism used by bacteria to sense and respond to their environment. The study shows that when the mechanism is disrupted, the giant surface protein is overproduced -- giving the cells a spiny, or "porcupine-like" appearance -- and the bacteria lose their ability to form clumps. Furthermore, the research team found that this clumping defect also makes the bacteria less dangerous in an experimental model of the serious staph infection, endocarditis.

For further details, see:

Jennifer N. Walker, Heidi A. Crosby, Adam R. Spaulding, Wilmara Salgado-Pabón, Cheryl L. Malone, Carolyn B. Rosenthal, Patrick M. Schlievert, Jeffrey M. Boyd, Alexander R. Horswill. The Staphylococcus aureus ArlRS Two-Component System Is a Novel Regulator of Agglutination and Pathogenesis. PLoS Pathogens, 2013; 9 (12): e1003819 DOI: 10.1371/journal.ppat.1003819

Posted by Tim Sandle

Saturday, 29 March 2014

Stressed Bacteria Time Lapse Movie

Sigma B is thought to be a major regulator in bacterial stress. Release of the protein can be triggered by many different and stressful environmental conditions such as extreme temperatures or starvation. Scientists studied the sigma B protein in bacterial cells. And found that when released sigma B can activate over 150 genes. In the short time lapse movie below the bacteria (B. subtilis) are modified to glow green when sigma B springs into action. During growth they are exposed to a stress causing chemical. Much as expected the sigma B protein is activated. But then, to our surprise also, the protein is quickly switched off again while the chemical is still present in the environment. Instead of shifting from one steady state to another steady state the sigma B regulatory protein is released in pulses. This definitely requires more research, but for now, it is a fascination short bacteria Time Lapse movie. Enjoy!


About the makers of this movie: Researchers at the California Institute of Technology have found a new kind of stress response in a bacterial species called B. subtilis. Instead of just shifting from one steady state to another and staying there, the cells deal with stress—such as the lack of food—by activating a regulatory protein in steady pulses. The cell cranks up the frequency of the pulses if there’s more stress. By attaching fluorescent proteins to the genetic circuit responsible for B. subtilis‘s stress response, the researchers observed each pulse as a green flash.

Posted by Tim Sandle

Friday, 28 March 2014

What Is The Gram stain?

What Is The Gram stain?

Tim Sandle
By Tim Sandle
The primary staining technique used to differentiate bacteria is the Gram stain. The Gram stain method employed includes the four-step technique: Crystal violet (primary stain); iodine (mordant); alcohol (decolorizer); safranin (counter stain) or the three-step method in which the decolorization and counter-staining step are combined. Done correctly, Gram-positive organisms retain the crystal violet stain and appear blue; Gram negative organisms lose the crystal violet stain and contain only the counter-stain safranin and thus appear red. Common pitfalls in this method are that heat fixation may cause Gram-positive cells to stain Gram-negative and older cultures may give Gram-variable reaction; using too much decolorizer could result in a false Gram-negative result and not using enough decolorizer may yield a false Gram-positive result.
The Gram reaction is based on the differences in the cell wall composition for the two cellular ‘groups'. The bacteria that retained the stain (the Gram-positive bacteria) have a higher peptidoglycan and lower lipid content than those that do not retain the stain (the Gram-negative bacteria). The effect of the solvent is to dissolve the lipid layer in the cell wall of the Gram-negative bacteria, thereby causing the crystal violet to leach out; whereas for Gram-positive bacteria the solvent dehydrates the thicker cell walls, blocking any diffusion of the violet-iodine complex, which closes the pores of the cell and retains the stain. There are now several automated Gram stain devices available on the market that can reduce the labour requirement required when performing several multiple Gram stains and, possibly, improve accuracy.
In addition to the difference based on cell wall, microscopic examination of the stains allows the cellular shape to be determined. Bacteria commonly fall into the categories of coccus (spherical), rod, vibrio (curved), spirilla (spiral) and plemomorphic (variable).
Tim Sandle, Ph.D, M.A., BSc (Hons), CBiol, MSBiol., MIScT – Dr. Sandle is the Head of Microbiology at the UK Bio Products Laboratory. Dr. Sandle is a chartered biologist and holds a first class honors degree in Applied Biology; a Masters degree in education; and obtained his doctorate from Keele University. His role involves overseeing a range of microbiological tests, batch review, microbiological investigation and policy development. In addition, he is an honorary consultant with the School of Pharmacy and Pharmaceutical Sciences, University of Manchester and is a tutor for the university's pharmaceutical microbiology M.Sc course. Dr. Sandle serves on several national and international committees relating to pharmaceutical microbiology and cleanroom contamination control (including the ISO cleanroom standards). He is currently chairman of the PharMIG LAL action group and serves on the NBS cleaning and disinfection committee. He has written over eighty book chapters, peer reviewed papers and technical articles relating to microbiology. He is currently the editor of the Pharmaceutical Microbiology Interest Group Journal and runs an on-line microbiology forum (www.pharmig.blogspot.com). Dr. Sandle is an experienced auditor and frequently acts as a consultant to the pharmaceutical and healthcare sectors.

Posted by Tim Sandle

Thursday, 27 March 2014

The influence of the microbiome on aging

We live with millions of microorganisms and these bacteria have a critical impact on our health and diseases. In addition to their effects on disease, studies have begun to suggest that microbes influence something much more intrinsic to their host — the aging rate.

In the current issue of Cell, a minireview highlights recent research showing that microbiota in invertebrates directly and indirectly influence the host genome to affect longevity. Beyond the presence or absence of bacteria, specific bacterial species have unique effects on host aging. (2014, Cell 156, 408.)

Posted by Tim Sandle

Wednesday, 26 March 2014

An overview of the options for antimicrobial surfaces in hospitals

An attractive option is to somehow make surfaces antimicrobial to exert a continuous reduction in the level of contamination. A recent review by Prof Hilary Humphreys provides a useful overview of the various approaches to antimicrobial surfaces.9 There are several approaches to making a hospital surface ‘antimicrobial’:
  • Permanently ‘manufacture in’ an agent with antimicrobial activity (e.g. copper or a chemical).
  • Periodically apply an agent with antimicrobial activity (e.g. copper containing liquid agents, or chemical disinfectants with residual activity).
  • Physically alter the properties of a surfaces to make it less able to support microbial contamination or easier to clean (e.g. antibiofilm surfaces).
Further to this, Jon Otter has written an informative article about antimicrobial surfaces. The review is published on the Micro Blog.

Posted by Tim Sandle

Tuesday, 25 March 2014

The influence of bacterial infection on host metabolism

Bacterial pathogens employ various strategies to interact with and subvert host defenses to promote survival and proliferation. Such subversion is often mediated by the specific interaction of bacterial effectors with host-encoded proteins and other molecules to modify host function.

In the current issue of Cell, scientists reported that the extracellular pathogenic bacterium group A Streptococcus (GAS) modulates host metabolism to regulate its own sensing and proliferation. GAS releases streptolysin toxins, leading to ER stress during adherence to host cells, which promotes the production of asparagine (ASN). The bacteria senses the released ASN, modifying the expression of about 17% of its genes.

Posted by Tim Sandle

Monday, 24 March 2014

Quality Management Systems

Quality Management Systems

Tim Sandle
By Tim Sandle, Ph.D, M.A., BSc (Hons), CBiol, MSBiol., MIScT
The objectives of a Quality Management System are to:
  • Set direction and meet customers' expectations
  • Improve process control
  • Reduce wastage
  • Lower costs
  • Increase market share
  • Facilitate training
  • Involve staff
  • Raise morale
The basis of a Quality Management System is set out in: ISO/FDIS 9001:2000 — Quality management systems — Requirements.
The adoption of a QMS needs to be a strategic decision of an organisation, and is influenced by varying needs, objectives, the products/services provided, the processes employed and the size and structure of the organisation. A QMS must ensure that the products/services conform to customer needs and expectations, and the objectives of the organisation. Issues to be considered when setting up a QMS include:
  • Design and build includes the structure of the quality management system, the process and its
  • implementation. It's design must be led by senior managers to suit the needs of the organisation, and this is ideally done using a framework to lead the thinking. Design of the QMS should come from determining the organisation's core processes and well-defined goals and strategies, and be linked to the needs of one or more stakeholders. The process for designing and building the QMS must also be clear, with the quality function playing a key role, but involvement and buy-in to the system must also come from all other functions.
  • Deployment and implementation is best achieved using process packages, where each core process is broken down into sub-processes, and described by a combination of documentation, education, training, tools, systems and metrics. Electronic deployment via Intranets is increasingly being used.
  • Control of the QMS will depend on the size and complexity of the organisation. ISO is a site-based system, and local audits and reviews are essential even if these are supplemented by central reviews.
  • Local control, where possible, is effective, and good practice is found where key stakeholders are documented within the process and where the process owner is allowed to control all of the process. Ideally, process owners/operators are involved in writing procedures.
  • Measurement is carried out to determine the effectiveness and efficiency of each process towards attaining its objectives. It should include the contribution of the QMS to the organisation's goals; this could be achieved by measuring the following:
    • Policy definition completeness
    • Coverage of business
    • Reflection of policies
    • Deployment
    • Usage
    • Whether staff find the QMS helpful in their work
    • Speed of change of the QMS
    • Relevance of QMS architecture to the job in hand
A form of scorecard deployed through the organisation down to individual objective level can be employed, and the setting of targets at all levels is vital.
Review of the effectiveness, efficiency and capability of a QMS is vital, and the outcome of these reviews should be communicated to all employees. Reviewing and monitoring should be conducted whether or not improvement activities have achieved their expected outcomes.
Improvement should follow as a result of the review process, with the aim of seeking internal best practice. It is part of the overall improvement activities and an integral part of managing change within the organisation.
An effective QMS must ensure that the organisation has a strong Customer Focus. Customer needs and expectations must be determined and converted into product requirements. Top management have to demonstrate Leadership. Providing unity of purpose through an appropriate quality policy, ensuring that measurable objectives are established, and demonstrating that they are fully committed to developing, sustaining and improving the QMS.
Managers must ensure that there is Involvement of People at all levels in the organisation. This includes ensuring that there is an awareness of the importance of meeting customer requirements and responsibilities in doing this, and people are competent, on the basis of appropriate training and experience.
An effective QMS must be a strategic tool designed to deliver business objectives, and must have, at its core, a Process Approach, with each process transforming one or more inputs to create an output of value to the customer. The key business processes may be supported by procedures and work instructions in those cases where it is judged necessary to rigidly define what rules are to be followed when undertaking a task. Most organisations will have core business processes that define those activities that directly add value to the product or service for the external customer, and supporting processes that are required to maintain the effectiveness of the core processes.
The understanding of the many interrelationships between these processes demands that a Systems Approach to management is adopted. The processes must be thoroughly understood and managed so that the most efficient use is made of available resources, to ensure that the needs of all the stakeholders — customers, employees, shareholders and the community — are met.
Customer satisfaction is a constantly moving entity depending on changes in technology and the market place, so an effective QMS must be in a state of Continual Improvement. For this to be achieved, attention needs to be given to both the voice of the customer - through complaint analysis, opinion surveys and regular contacts – and the voice of the processes – through measurement, monitoring and analysis of both process and product data. This will result in Factual Decision Making.
Each organisation is itself only a link in the chain of a larger raw material process, and for the long term needs of the community and the organisation there needs to be Mutually Beneficial Supplier Relationships.
Audits, reviews and assessments

A good QMS will not function or improve without adequate audits and reviews. Audits are carried out to ensure that actual methods are adhering to the documented procedures, whilst system reviews should be carried out periodically and systematically, to ensure the system achieves the required effect.
There should be a schedule for carrying out audits, with different activities possibly requiring different frequencies. An audit should not be conducted just with the aim of revealing defects or irregularities – they are for establishing the facts rather than finding faults. Audits do indicate necessary improvement and corrective actions, but must also determine if processes are effective and that responsibilities have been correctly assigned. The emphasis on process improvement and enhancing customer satisfaction in the revised standard will require a more thoughtful approach to auditing.
The generic steps involved in an audit are:
  • Initiation
    • Scope
    • Frequency
  • Preparation
    • Review of documentation
    • The programme
  • Working documents
    • Execution
  • Opening meeting
    • Examination and evaluation
    • Collecting evidence
    • Observations
  • Close the meeting with the auditee
  • Report
    • Preparation
    • Content
    • Distribution
    • Completion
    • Report
    • Submission
    • Retention
A quality management system review should take place, possibly once a year, which should cover:
  • Results of audits
  • Customer feedback
  • Process and product conformity
  • Status of preventative and corrective actions
  • Follow up actions from previous management reviews
  • Changes that could effect the QMS
  • Recommendations for improvements
  • Outputs should include:
  • Improvements to the QMS and processes
  • Improvements of a product related to customer requirements
  • Resource needs
In addition, the procedures for conducting audits and reviews and the results from them should be documented, and also be subject to review. Internal system audits and reviews should be positive and conducted as part of the preventative strategy, and not as a matter of expediency resulting from problems. The assessment of a quality system against a standard or set of requirements by internal audit and review is known as a first-party assessment or approval scheme. If an external customer makes the assessment of a supplier, against either its own, or a national or international, standard, a second-party scheme is in operation. The assessment by an independent organisation, not connected with any contract between the customer and supplier, but acceptable to them both, is an independent third-party assessment scheme.
All managers, not just the staff in the "quality department", need to be fully committed to operating an effective quality management system for all the people within the organisation. The system must be planned to be effective and achieve its objectives in an uncomplicated way. It should also not be static, but be flexible, to enable constant seeking of improvements
Quality Management Review

The object of the Quality Management Review is to provide an overview of the results from the Quality Control and Quality Assurance functions. The review forms part of an organization's Quality Management Systems (as detailed in ISO 9001). The issues covered will vary between different workplaces. However, issues often included issues are results of audits; discussions of follow up items; a review of progress relating to non-compliances and discussions relating to the effectiveness of corrective and preventative action; test and sampling efficiency times; customer complaints; rejects; out of specification results from test data; microbiological contamination issues and trends in relation to processing of product and to the manufacturing environment. Issues of major concern are often considered using risk management techniques.
Data is presented by participants from different departments. The data is normally measured against a target (to allow for effectiveness, efficiency and capability to be measured). This can be relatively straightforward, such as the number of internal audits completed against a target level (such as 95% of audits to be completed to schedule) or more complex when relating to statistical process control charts for laboratory test data.
Participants will include representatives of laboratories, production, process, pharmacy and ward management. Meetings are held either monthly, quarterly or annually (at my workplace, the meetings are held quarterly).
Where problems or upward trends are identified the aim of the meeting is to identify the risk, assign a priority, to agree actions and to track progress. For issues that cannot be satisfactorily dealt with these are referred to senior management. Quality Management Review meetings may sometimes elect to form subgroups to examine specific areas, such as sampling; new processes technologies and so on.
It is important that the outcomes of the Quality Management Review are communicated to all members of staff.
Tim Sandle, Ph.D, M.A., BSc (Hons), CBiol, MSBiol., MIScT – Dr. Sandle is the Head of Microbiology at the UK Bio Products Laboratory. Dr. Sandle is a chartered biologist and holds a first class honors degree in Applied Biology; a Masters degree in education; and obtained his doctorate from Keele University. His role involves overseeing a range of microbiological tests, batch review, microbiological investigation and policy development. In addition, he is an honorary consultant with the School of Pharmacy and Pharmaceutical Sciences, University of Manchester and is a tutor for the university's pharmaceutical microbiology M.Sc course. Dr. Sandle serves on several national and international committees relating to pharmaceutical microbiology and cleanroom contamination control (including the ISO cleanroom standards). He is currently chairman of the PharMIG LAL action group and serves on the NBS cleaning and disinfection committee. He has written over eighty book chapters, peer reviewed papers and technical articles relating to microbiology. He is currently the editor of the Pharmaceutical Microbiology Interest Group Journal and runs an on-line microbiology forum (www.pharmig.blogspot.com). Dr. Sandle is an experienced auditor and frequently acts as a consultant to the pharmaceutical and healthcare sectors.

Posted by Tim Sandle

Sunday, 23 March 2014

What do we mean by ‘cleaning’ and ‘disinfection’?

Dr Sally Bloomfield has written an informative, introductory article about cleaning and disinfection. Here is an extract:

"Some people work on the basis that “clean” means visibly clean, and “microbiologically safe clean” means a chemical or thermal disinfectant has been used.  But then how can we communicate that hand washing can make hand surfaces microbiologically safe” without need for a disinfectant.  There is a notion that “cleaning” is hygienically inferior to disinfection – but data now shows that the log reduction by handwashing with soap can be equivalent to that achieved by alcohol handrubs if done properly, and you have access to running water.  We put much effort into hand hygiene compliance, but relatively little into stressing that handwashing technique to deliver hands which are “fit for purpose” is equally important."

To read the full article, go to Micro Blog.

Posted by Tim Sandle

Saturday, 22 March 2014

Safety Data Sheet (SDS) and laboratory management

The average laboratory typically has thousands of materials on site and each one requires a Safety Data Sheet (SDS). In addition, those SDSs are frequently updated by the supplier, meaning that the laboratory manager must keep track of not only where each SDS is and which SDS need updating, but also any new SDSs that need to be attached to new chemicals being ordered by the lab. Keeping up with those changes can be a complex and time-consuming task.

To assist laboratory users, Accelrys have produced a white paper on the control and management of SDSs. For further details, click here.

Posted by Tim Sandle

Cleaning and Disinfection of Cleanrooms


Maintaining environmental control in a pharmaceutical manufacturing environment is  in large part dependent on the facility‟s cleaning and disinfection program. The  program requires the selection of the appropriate disinfectants, their proper  application, and validation of their capability to inactivate vegetative cells.

In relation to this important subject, Tim Sandle has written a paper for AsepTech, outlining the key considerations for the cleaning and disinfection of cleanrooms.

The paper can be accessed here.

Posted by Tim Sandle

Friday, 21 March 2014

Use Of Hydrogen Peroxide Vapor Biodecontamination Technology


White Paper: Use Of Hydrogen Peroxide Vapor Biodecontamination Technology For Facility Commissioning.

Arthur Papineau, BS ChE, MBA and John Klostermyer, PhD, STERIS Life Sciences have written a paper on the use of biocontamination technology.

The abstract reads:

“This article discusses the successful use of hydrogen peroxide vapor in the biodecontamination of a new 100,000+ ft3 lab animal research facility for the purpose of a facility commissioning process. It will discuss the reasons why this process was chosen; the preparation of the facility; the execution of the biodecontamination; and the results.

There are many methods of biodecontamination available on the market today. Each method has its advantages and disadvantages depending upon the scope of the project and desired level of bioburden reduction. In this particular case, a very high standard was set. The Customer wanted to have the entire 100,000+ ft3 facility and equipment within it (i.e. computers, washers, cage racks, etc.) biodecontaminated with a validatable process that delivered a wide range of efficacy, a high level of material compatibility, the least amount of downtime possible, and a minimum of a 6-log reduction in bioburden.”

To access the article, go to Pharmaceutical Online

Posted by Tim Sandle

Thursday, 20 March 2014

Spotlight on pharmaceutical microbiology: Tim Sandle interview

As part of a feature on pharmaceutical microbiology, Tim Sandle has been interviewed by Pharmaceutical Bioprocessing. The interview appears in the current edition (Vol. 2, Issue 1).

Tim Sandle is Head of Microbiology at Bio Products Laboratory (Hertfordshire, UK). In addition, he is a visiting tutor with the School of Pharmacy and Pharmaceutical Sciences, University of Manchester (Manchester, UK) where he teaches the University’s pharmaceutical microbiology MSc course. Sandle serves on several national and international committees including the microbiology society Pharmig (Hertfordshire, UK). Sandle has written almost three hundred book chapters, peer-reviewed papers and technical articles relating to microbiology, as well as being the author or co-editor of eight books. Sandle runs a microbiology discussion site, Pharmaceutical Microbiology. Sandle spoke to Pharmaceutical Bioprocessing for the first in a series of interviews on pharmaceutical microbiology.

To read the interview, go to Pharmaceutical Bioprocessing.

Wednesday, 19 March 2014

Combating bacterial infections

Scientists have identified a chemical substance with the potential of acting as a new drug to treat bacterial infections. This becomes important as the rate of antibiotic resistance increases.
A research team has identified a chemical substance with the potential of acting as a new drug to treat bacterial infections, particularly urinary tract infections. Unlike antibiotics the new drug does not destroy pathogenic bacteria; instead it disarms them.
This is based on the main way that pathogenic bacteria infect human cells. Many pathogenic bacteria attach to a cell before they can infect it with the aid of hair-like structures called pili. Pili connect a bacterium to another of its species, or to another bacterium of a different species, and build a bridge between the interior of the cells. Dozens of these structures can exist on the bacteria. Knowing this, the research team screened databases of chemical components in search of chemical molecules that could inhibit pili formation.

This is illustrated in the following video:


The search was successful and the scientists found an inhibitor that interferes with an essential step in the assembly process of the pili. They tried the drug candidate out on a pathogenic form of Escherichia coli that causes urinary tract infections. This worked and the E. coli were unable to attach themselves to the host cells.
The research was led by Alvin Lo and Han Remaut (VIB/Vrije Universiteit Brussel). The findings have been published in the Journal of Antimicrobial Chemotherapy. The research paper is titled “Suppression of type 1 pilus assembly in uropathogenic Escherichia coli by chemical inhibition of subunit polymerization.”

Posted by Tim Sandle

Tuesday, 18 March 2014

Stem cells must be regulated as a drug product

A US federal appeals court maintains that stem cells proliferated in a lab must be regulated as a drug. The D.C. Circuit Court of Appeals has ruled that culturing a patient’s stem cells for therapeutic use falls under the aegis of the US Food and Drug Administration (FDA). FDA has said therapeutic stem cells should be regulated as drugs.

In relation to this topic, Leigh Turner, a bioethicist at the University of Minnesota, told Nature News that: "It is much too simplistic to think that stem cells are removed from the body and then returned to the body without a ‘manufacturing process’ that includes risk of transmission of communicable diseases. Maintaining the FDA’s role as watchdog and regulatory authority is imperative."

Posted by Tim Sandle

Monday, 17 March 2014

Studying the bacteria inside buildings

A survey of bacteria in a University of Oregon building reveals that architecture influences the indoor microbiome. Patterns vary by age, space, locale and, surprisingly, architecture.

Different aspects of a building’s design and use, including the number of doors per room, its average occupancy, and the way air circulates, can each influence the makeup of the bacterial communities that occupy buildings, according to a study published in the journal PLOS One (titled "Architectural Design Drives the Biogeography of Indoor Bacterial Communities").

The research was undertaken by scientists based at the University of Oregon’s Biology and the Built Environment Center. For their study, the scientists vacuumed up dust samples from around the university’s Lillis Hall, then analyzed bacterial DNA from the samples to estimate microbial diversity.

Talking to Popular Science, lead researcher Jessica Green said: “What we did in our paper was ask this really fundamental question. Given this hypothesis that is becoming more and more actualized—that the indoor microbiome is important to health—do we have any control over what kind of microbes are indoors or out?”

The researchers identified nearly 33,000 different major groups of bacteria in its survey, with different room types comprising a unique microbial composition. Known gut bacteria were common in bathrooms, for example, while offices with windows tended to have more soil- and plant-associated bacterial species.

The impact of these diverse indoor microbiomes on human health is a matter of debate. However, since this is a new area of microbiology, more data collected over time will reveal the impact of 'building bacteria'. Hospitals, for example, are likely to be interested in what bacteria are populating their rooms and hallways, according to the science website Quartz.
Posted by Tim Sandle

USP Pharmacopeial : 40(1) In-Process Revision



Future planned revisions to the USP include:
  • 51 Antimicrobial Effectiveness Testing (USP38-NF33)
  • 209 Low Molecular Weight Heparin Molecular Weight Determinations [NEW] (USP38-NF33)
  • 251 Lead (USP38-NF33)
  • 660 Containers - Glass (USP38-NF33)
  • 852 Atomic Absorption Spectroscopy [NEW] (USP38-NF33)
  • 85> Fluorescence Spectroscopy [NEW] (USP38-NF33)
  • 854 Mid-Infrared Spectroscopy [NEW] (USP38-NF33)
  • 857 Ultraviolet-Visible Spectroscopy [NEW] (USP38-NF33)
  • 1852 Atomic Absorption Spectroscopy - Theory and Practice [NEW] (USP38-NF33)
  • 1853 Fluorescence Spectroscopy - Theory and Practice [NEW] (USP38-NF33)
  • 1854 Mid-Infrared Spectroscopy - Theory and Practice [NEW] (USP38-NF33)


According to the USP:

“This section contains proposals for adoption as official USP or NF standards (either proposed new standards or proposed revisions of current USP or NF standards). These may be any of the following: (1) proposed revisions placed directly under In-Process Revision, or (2) modifications of revisions previously proposed under In-Process Revision. Readers should review material in this section and provide comments to the Scientific Liaison using the contact information appearing at the end of each proposal. Information on how to comment can be found under the "Participation" section of www.usp.org. It is important to send comments promptly, using the comment deadline listed after each title."

The U.S. Pharmacopeial Convention (USP) is a scientific nonprofit organization that sets standards for the identity, strength, quality, and purity of medicines, food ingredients, and dietary supplements manufactured, distributed and consumed worldwide. USP’s drug standards are enforceable in the United States by the Food and Drug Administration, and these standards are used in more than 140 countries.
Posted by Tim Sandle

Sunday, 16 March 2014

Llama antibodies can tackle Clostridium difficile

Highly specific antibodies derived from llamas may provide a new method for controlling deadly infections from the opportunistic bacterial pathogen Clostridium difficile, according to a new report.
The report describes how llama antibodies recognize the Clostridium difficile’s two central toxin proteins -- toxin A (TcdA) and toxin B (TcdB). Clostridium difficile (C diff) is a species of Gram-positive spore-forming bacteria that is best known for causing antibiotic-associated diarrhea. C. difficile infection is a growing problem in healthcare facilities. Outbreaks occur when humans accidentally ingest spores in a medical facility.
To show that the llama antibodies had the best structure to combat C diff, the scientists studied the three-dimensional structures of antibody-toxin complexes using X-ray crystallography.

 
From this, studies were undertaken to show that the smaller size of the llama antibodies had the greatest potential against C diff. Further trials are set to continue.
The implications of the research are that a new generation of engineered antibodies could be created and that these may be more effective at preventing the toxins from damaging the intestine during the normal course of the disease.
The research was undertaken at the Alberta Glycomics Centre at the University of Calgary and the University of Alberta, in collaboration with researchers at the National Research Council of Canada in Ottawa. The findings have been published in the Journal of Biological Chemistry. The research paper is titled “Structural Basis for Antibody Recognition in the Receptor-binding Domains of Toxins A and B from Clostridium difficile.”

Posted by Tim Sandle

The A-Z of HPAPIs



High potency active pharmaceutical ingredients (HPAPIs) is fastest growing segment of pharmaceutical industry.

BioPharma have put together a handy A-Z of everything you need to know about High Potency Active Pharmaceutical Ingredients. The guide can be accessed here: HPAPIs.

Posted by Tim Sandle

Saturday, 15 March 2014

A pathogenic pathway — the influence of bacterial infection on host metabolism

Bacterial pathogens employ various strategies to interact with and subvert host defenses to promote survival and proliferation. Such subversion is often mediated by the specific interaction of bacterial effectors with host-encoded proteins and other molecules to modify host function.

In the current issue of Cell, scientists reported that the extracellular pathogenic bacterium group A Streptococcus (GAS) modulates host metabolism to regulate its own sensing and proliferation. GAS releases streptolysin toxins, leading to ER stress during adherence to host cells, which promotes the production of asparagine (ASN). The bacteria senses the released ASN, modifying the expression of about 17% of its genes. (2014, Cell 156, 97.)

Posted by Tim Sandle

Friday, 14 March 2014

Antibiotic ‘smart bomb’ hits out at pathogens


Microbiologists have developed an antibiotic "smart bomb" that can identify specific strains of bacteria and sever their DNA, eliminating the infection. This could help to combat antibiotic-resistant bacteria.
There are two problems with antibiotics. First, many bacteria are developing resistance (to the degree that this has become an issue of international concern). Second, they kill indiscriminately, eliminating good and bad bacteria. By killing beneficial bacteria, antibiotics can be harmful by disrupting the digestive system and other workings of the body.
To overcome this, a science group has developed a new compound that can target pathogens while leaving "good" bacteria untouched. The new approach works by taking advantage of a part of an immune system present in many bacteria called the CRISPR-Cas system (a short version of Clustered Regularly Interspaced Short Palindromic Repeats). The CRISPR-Cas system protects bacteria from invaders such as viruses.
The drug developed by the scientists is able to target the CRISPR-Cas system in specific bacteria. The drug then proceeds to cut the bacterial DNA into pieces, thereby killing the bacterium. The compound can be developed to target specific pathogens like Salmonella.
The study was carried out at North Carolina State University and the findings have been published in the journal mBio. The paper is titled “Programmable Removal of Bacterial Strains by Use of Genome-Targeting CRISPR-Cas Systems.”

Posted by Tim Sandle

Counterfeit Medicines and Similar Crimes (free publication)


The EDQM has just launched a new webpage dedicated to its publications in the field of counterfeit medicines and similar crimes. The publications cover a variety of topics including risk communication, facts and practical advice on dealing with the problem, and outline strategies for assisting authorities to revise legislation on medical products, public health and criminal law in order to better protect public health.

For further details see: EDQM

Posted by Tim Sandle

Thursday, 13 March 2014

Architecture influences the indoor microbiome

Various aspects of a building’s design and use, including the number of doors per room, its average occupancy, and the way air circulates, can influence the makeup of the bacterial communities that occupy the space, according to a study published in PLOS One - 'Architectural Design Drives the Biogeography of Indoor Bacterial Communities'.

The research comes from biologists and architects at the University of Oregon’s Biology and the Built Environment Center, who vacuumed up dust samples from around the university’s Lillis Hall, then analyzed bacterial DNA from the samples to estimate microbial diversity.

The team identified nearly 33,000 different major groups of bacteria in its survey, with different room types comprising a unique microbial composition. Known gut bacteria were common in bathrooms, for example, while offices with windows tended to have more soil- and plant-associated bacterial species.

Posted by Tim Sandle

Wednesday, 12 March 2014

New rapid method to detect Salmonella

A novel biosensor developed by scientists at Rice University in collaboration with colleagues in Thailand and Ireland may make the detection of pathogens much faster and easier for food-manufacturing plants.

The method consists of an array of tiny ‘diving boards’ that can identify many strains of Salmonella at once.
The "diving boards" are a set of microcantilevers, each of which can be decorated with different peptides that have unique binding affinities to strains of the salmonella bacteria. When a peptide catches a bacterium, the cantilever bends ever so slightly, due to a mismatch in surface stress on the top and bottom. A fine laser trained on the mechanism catches that motion and triggers the alarm. The system is sensitive enough to warn of the presence of a single pathogen.

For further details, see the following paper:

Jinghui Wang, M. Josephine Morton, Christopher T. Elliott, Nitsara Karoonuthaisiri, Laura Segatori, Sibani Lisa Biswal. Rapid detection of pathogenic bacteria and screening of phage-derived peptides using microcantilevers. Analytical Chemistry, 2014; : 140113115511000 DOI: 10.1021/ac403437x Posted by Tim Sandle

Tuesday, 11 March 2014

Researchers Launch Phase 1 Trial of Potential MRSA Treatment

NIAID-funded researchers have launched a Phase 1 clinical trial of the candidate oral antibiotic EDP-788, a drug intended to treat methicillin-resistant Staphylococcus aureus (MRSA) infections. The trial, conducted by biotechnology company Enanta Pharmaceuticals through a contract with NIAID, will evaluate the drug’s safety and how it is broken down and processed in the body.

For more information on EDP-788, MRSA, and the new clinical trial, see the NIAID.

Posted by Tim Sandle

Monday, 10 March 2014

Long-acting HIV drug shows promise

Two new sets of research show that regular injections of an antiretroviral drug protects macaques monkeys from simian-human immunodeficiency virus infection. The research has raised hopes that preventing HIV infection in humans will soon be as easy as a shot in the arm four times a year. Researchers are optimistic that the findings could one day lead to a new generation of HIV drugs for infected people.

The medication has been developed by researchers from Rockefeller University, GlaxoSmithKline, and the Tulane National Primate Research Center. The new drug is termed a "integrase strand-transfer inhibitor GSK744", and it has been developed as a long-acting injectable, designed to be administered every three months.

GSK744 (also known as S/GSK1265744) is an investigational new drug under development for the treatment of HIV infection. Integrase inhibitors are a class of antiretroviral drug designed to block the action of integrase, a viral enzyme that inserts the viral genome into the DNA of the host cell. Integrase inhibitors were initially developed for the treatment of HIV infection, but they could also be applied to other retroviruses. GSK744 works by interfering with the enzyme that HIV uses to insert its DNA into the human genome. This blocks the key step needed for the AIDS virus to replicate itself, and the viral DNA simply degrades inside the cell.

In investigational studies, the GSK744 agent has been packaged into nanoparticles (GSK744LAP) conferring an exceptionally long half-life of 21–50 days following a single dose. In theory, this would make possible suppression of HIV with dosing as infrequently as once every three months. Animal studies indicate that this theory is getting closer to reality.

Based on a report in the journal Science, the drug has led to monkeys, infected with a virus very similar to HIV called simian-human immunodeficiency virus (SHIV), becoming protected. The study, by Chasity Andrews and colleagues is titled " Long-Acting Integrase Inhibitor Protects Macaques from Intrarectal Simian/Human Immunodeficiency Virus".

For the study, Andrews and her colleagues studied sixteen rhesus macaques. The monkeys were challenged with SHIV. After this,  eight monkeys received placebo and eight were administered with GSK744 (where the drug was injected intramuscularly). All eight macaques who received GSK744 remained averimic (that means without SHIV), during both the rectal challenge phase and during the subsequent follow-up ten-week examination phase. In contrast, all eight macques who received placebo were infected with SHIV during the challenge phase.

For the next phase of the study, the researchers aim to carry out similar tests where they will determine the minimum protective dose of GSK744 that is required in order to stop SHIV from replicating and multiplying.

Remarkably, a second research team from the U.S. Centers for Disease Control and Prevention (CDC) have found a similar effect when studying GSK744 in female monkeys.

Living with HIV can be a lonely and worrying time. One way of managing the illness is through meeting people going through the same condition and having the same worries. This is where a site like Positive Singles comes in. The site matches up people with STIs for friendship and romance.

With the new HIV research findings, both research groups have presented their results to the U.S. Centers for Disease Control and Prevention (CDC) annual Conference on Retroviruses and Opportunistic Infections, which held in Boston this week.


Commenting on the research findings, The New York Times explains: “If the findings can be replicated in humans, they have the potential to overcome a major problem in AIDS prevention: that many people fail to take their antiretroviral pills regularly."

In short, this could be the first step towards an HIV vaccine. In the meantime, ‘pre-exposure prophylaxis’ with drugs used to treat the infection has become one of the most promising strategies to cut down HIV infection rates among high-risk populations. On this basis the newly developed drug for monkeys could, within a short space of time, be turned into a long-acting injectable drug for people and one that could improve upon current treatments. Furthermore, since integrase inhibitors target a distinct step in the retroviral life cycle, they may be taken in combination with other types of HIV drugs to minimize adaptation by the virus. They are also useful in salvage therapy for patients whose virus has mutated and acquired resistance to other drugs.

The main obstacle for realizing a version effective against HIV is likely to be keeping up the treatment regime.  Philip Johnson of the Children’s Hospital of Philadelphia in Pennsylvania told the science site Science Now. “This is going to require multiple injections over the lifetime of an individual,” he said, asking: “How feasible is it truly in the long term?”

Much more likely it seems that having to make multiple pills each day for HIV. GSK744 injections have the potential to overcome a major problem in AIDS prevention: that many people fail to take their antiretroviral pills regularly.

Posted by Tim Sandle

New way to treat prion diseases


A research development reveals natural mechanism in brain cells that protects against “molecular piracy” of deadly neurological diseases.

Prion diseases lead to incurable neurodegenerative disorders such as Creutzfeldt-Jakob disease in humans, mad cow disease (bovine spongiform encephalopathy) and chronic wasting disease in deer and elk.

Research, published in The Journal of Clinical Investigation, stemmed from two largely unexplained factors for prion infections: their long incubation periods of up to five decades, and a noted time discrepancy between the accumulation of the misfolded rogue prion protein in the brain and when the disease actually shows symptoms.

In probing these mysteries, Westaway, U of A post-doctoral fellow Charles E. Mays, associate professor Jiri Safar of Case Western Reserve University and other international collaborating researchers began two years ago to study a molecule called the “shadow” of the prion protein. Significant changes in this shadow protein have directed researchers to examine the normal prion protein itself.

For further details, see the paper “Prion disease tempo determined byhost-dependent substrate reduction”.

Posted by Tim Sandle

Addressing antibiotic resistance

Humans face the very real risk of a future without antibiotics. The implications of this are that life expectancy could fall due to people dying from diseases that are readily treatable today. One concern for the future is the re- emergence of diseases that are ‘officially’ extinct, such as tuberculosis (a disease caused by Mycobacterium tuberculosis). The other concern is that without effective antibiotics, human society will no longer be able to conduct the types of medical procedures that can lead to immunosuppression. Such therapies include those for cancer treatments or to address autoimmune disorders.



With regard to this serious issue, Tim Sandle has written a review article for the journal Microbiology & Infectious Diseases. The article can be accessed on-line here: SOJ Microbiology and Infectious Diseases or, alternatively, contact Tim Sandle.

Sandle, T. (2014) Novel Methods to Address Antimicrobial Resistance, SOJ Microbiology & Infectious Diseases, 2 (1): 2-3

Posted by Tim Sandle