The American Medical Association (AMA) will be considering a resolution at their House of Delegates meeting in Chicago on June 13-17 to address the role of clothing and accessories (neckties, jewelry, wrist watches, etc.) in the spread of pathogens in hospitals. Resolution 720 advises that hospitals adopt dress codes that “minimize transmission of nosocomial infections, particularly in critical and intensive care units.”

The resolution also specifically refers to the United Kingdom’s “bare below the elbows” policy.  If passed, such a resolution would cause a major fashion change in hospitals — no more long sleeve white lab coats. The proposed resolution is expected to help healthcare workers be more compliant with hand washing guidelines, as there will be less concerns about getting jewelry, wrist watches and long sleeves wet.  Avoiding wet shirt sleeves and wrist jewelry is one of the driving forces behind using alcohol-based gels. What have you heard from your customers reagarding this resolution?  If passed, do you think this will change the demand for alcohol-based hand sanitizers?

We work in a customer service environment dealing with the in-vitro efficacy evaluation of many and varied antimicrobial products. One of the most important considerations in almost every study that we perform is proving effective neutralization/inactivation of the antimicrobial activity of the product(s) to be tested. In brief, proof of neutralization is necessary to ensure the validity of the test method. Without verification of neutralization, differentiation between –cidal (kill) or inhibitory activity is difficult, at best. And since most competitive marketing of products is based upon claims of “kills 99.9% of germs in 15 seconds” or “reduces bacteria by 99.99% in 10 minutes,” proof of neutralization is as important as the results of the bactericidal test itself.

Depending upon the active ingredient(s) of a test product, neutralization is accomplished chemically through the addition of neutralizing agents (i.e., lecithin, Tween 80, catalase, etc.), by dilution of the active ingredient to a sub-inhibitory or sub-lethal level, or by a combination of the two. One of the first questions asked of a potential customer is “what is the active ingredient of your product?” Without an accurate answer, the study design is a “best guess” – which neutralizing formula will work best to immediately inactivate a given product?

ASTM E 1054-08, Standard Test Methods for Evaluation of Inactivators of Antimicrobial Agents, provides excellent guidance on methodologies for demonstration of adequate neutralization. All neutralization verification procedures should, at minimum, include the following:

1) An arm/phase that provides data demonstrating that the neutralizing fluid immediately and completely neutralizes the product. If neutralization is not complete or immediate, residual “killing” activity may occur – a bacterial reduction reported for a 15 second exposure time may actually be a reduction for 30 seconds, 1 minute, or longer. If this happens, the danger is that a product may be reported as being more effective than it actually is.

2) An arm/phase demonstrating that the neutralizing fluid itself is not toxic to the organism(s) tested. If the neutralizer exhibits some antibacterial activity, it may actually “add to” the apparent antimicrobial activity of a product. Again, the danger is that a product may be reported as being more effective than it actually is.

3) An arm/phase providing data showing the starting bacterial challenge population. The data from this phase/arm are the basis of comparison for the previous phases – in general, neutralization procedures should employ a LOW population of bacteria, in terms of CFU/mL. If a challenge population is too high, a neutralizing system may appear to be effective, when in fact it is not.

4) REPLICATION – this allows for a statistical comparison of the data – PROOF that the neutralizing system is effective.

Verification of neutralization must be performed for all in-vitro efficacy evaluations of antimicrobial products to ensure the validity of the data; depending upon the type of evaluation and the familiarity with the active ingredient(s), this verification may be conducted in advance of a efficacy evaluation, or concurrent with the efficacy evaluation — as long as it is conducted using the same test formulation.

– Terri Eastman, Manager of the In Vitro Laboratory

 And no … I am not just saying this because upper management will read this. I am one of the fortunate people that have a job to say “I love”. If you can say, “I hate my job” … CONGRATULATIONS … you are still employed. 

As a clinical research manager conducting research on a vulnerable population particular attention must be given to the process of informed consent. This additional attention is required because vulnerable populations are populations who have limitations regarding their ability to provide informed consent and are more susceptible to coercion or undue influence. Because the decision to participate in clinical research must be made by a competent individual who has received the necessary information, and who has understood that information, special care must be taken to resolve language and comprehension barriers between the researchers and their potential subjects.

Therefore the researchers must set aside adequate time and resources to insure that the proper language, or languages, is used to inform the potential subjects of the requirements of informed consent. Not only should the language itself be presented at a comprehension level easily understood, but the content and concepts of the message must also be presented at a level that will allow their comprehension. Particular attention should be given to the concepts of clinical research, randomization, and inactive medications (placebo) due to complexity of the issues, their importance, and the relative inability of the normal population to completely comprehend them.

Each of these issues requires the attention of the Institutional Review Board (IRB). It is the responsibility of the IRB to determine that the minimal requirements of 45 CFR 46.111 are achieved, that additional safeguards for this vulnerable population are in place and that proper informed consent is obtained from each subject, prior to protocol approval.

Private, for-profit IRBs can function without ethical conflicts as long as their members are not directly involved in the research and the outcome of the review in no way benefits them as individuals.

John Dyba, Senior Account Executive

There has been a tremendous buzz in the first four months of 2009 about using social media to drive business.  For sales and marketing professionals, the question is “with so many social media recommendations, how does one know where to spend their time effectively?”  It seems like social media can easily overwhelm us.

After reading several articles on the topic, there does seem to be a collective opinion on how to develop a social media strategy.  First, start small and put your effort where it will get the most results.  Look at the social media sites where your customers are spending their time.  Is it Facebook, Linked-in, Twitter or something else?  Second, test one social media site/tool at a time and track results.  Just like conducting a scientific experiment, develop purpose, approach and key metrics.  Third, make the social media strategy compliment the search strategy.  Choose social media sites and tools that give search results for keywords related to your products or services.  This can be tested for each site as you add content to it.  Finally, position yourself as the “expert authority” in the market niche you serve.  Pass along helpful news and tips from industry organizations.  Create a question that is currently relevant, answer it, and encourage others to respond.

There is much to learn about the power of social media.  The strategies and tips undoubtedly go beyond these.   How are you using social media at your company?

Jim Verzuh, Director of Marketing.

In the Clinical Laboratory, we employ numerous methods to sample microorganisms from the skin of human volunteers. These sampling methods are not normally part of a microbiologist’s college curriculum, and newly hired technicians need to be trained appropriately to perform the methods in our laboratories. Two of the methods primarily employed at BioScience Laboratories, Inc. (BSLI) are the Glove-Juice Sampling Procedure and the Cylinder-Sampling Technique (also sometimes call the Cup-Scrub Method). In addition to proper training, we also employ statistical techniques to help identify those requiring additional training and for improving upon our methods.

For studies that involve evaluating Handwash products, we employ the Glove-Juice Sampling Procedure to sample microorganisms from the hands. A sterile glove is placed over the hand and a measured volume of sampling fluid is placed in the glove. The method of gloving can differ depending on the type of study. Hands may be wet, dry, held higher than elbow, or held lower than elbow. The glove is secured at the wrist using a tight-fitting elastic band. The hand is then massaged in a standardized manner for 1 minute by the technician. Following the massage, a sterile pipette is used to draw out a volume of fluid that is then diluted and plated. As you can see, the multitude of steps would have to be practiced and standardized so that all technicians perform the procedure in the same manner, and mistakes that may cause the loss of data are not made.

Sampling microorganisms from a small area of skin is performed using the Cylinder-Sampling Technique. A small (~1 in diameter), stainless steel cylinder is held tightly on the skin with one hand, while the other hand uses a pipette to place a small measured volume of fluid into the cylinder. A rubber-tipped glass rod is then rubbed against the skin inside the cylinder for 1 minute using a standardized, sweeping motion. A new pipette is used to transfer the fluid to a test tube. The procedure is repeated, with the second volume of fluid being removed and pooled with that fluid from the first sample. In addition to being trained to perform the procedure, technicians also need to learn the importance of cylinder placement and how to deal with sampling at a multitude of anatomical sites. This technique, too, requires lengthy training and practice.

Over my tenure at BSLI, Dr. Paulson has always employed statistical methods, such as Exploratory Data Analysis, to “look” at the data to determine and identify data points that are extreme “outliers” compared to the other data points. This statistical technique is used to identify human volunteers who may have not followed product restrictions or those who did not perform a wash procedure correctly, as well as help to determine if a noted incident that occurred had an affect on the data. EDA can also be used to evaluate the sampling methods and the technicians performing them… that is, identifying technicians who may require additional training. Conversely, EDA can also be used to identify technicians who perform the procedure well. The sampling method then can be adjusted to how they perform it, thereby decreasing variability and increasing the acuity of the conclusions drawn from the data.

Properly training technicians at BioScience Laboratories, Inc., is always a priority. We understand that our methods are non-traditional, but data generated by them is important to evaluating the true efficacy of a product. The initial training of a technician is vital to the conductance of the study, but continually monitoring the technicians’ performance and identifying minor adjustments has been and always will be a continual process here at BioScience Laboratories, Inc.

Christopher M. Beausoleil, CCRP

 Historically, the effectiveness of virus inactivation is estimated using procedures for detecting the loss of the virus’ ability to replicate. Until recently, this approach could not be applied to the majority of noncultivable viruses. There have been several attempts to develop indirect infectivity assays in order to solve the problem. One attempt is the enzymatic pretreatment of inactivated viruses with proteinase K and Rnase . Different research groups presented contradictory results on the reliability of this method. For the most part, it is because the effectiveness of the enzymatic pretreatment depends on the type of inactivation producing different levels of disruption, and hence, different levels of accessibility of peptide bonds to proteinase. Moreover, enzymatic pretreatment represents a secondary inactivation of presumably inactivated viruses, which might further enhance virus disruption.  Relatively reliable results were obtained by assessing loss of the inactivated virus’ ability to bind cellular receptors. The cell attachment capabilities of Poliovirus, Feline Calicivirus and HAV were lost in response to high temperature and hypochlorite treatment. However, UV inactivation did not prevent HAV from binding to the cellular receptor. This indicates that alterations of virus cell-binding sites are not the only mechanism of inactivation.   As a whole, detection of virus viability remains essential. The infectivity assay by means of cell culture has been the gold standard against which all new technologies for virus detection are evaluated. Methodologies combining cell culture and molecular techniques – for example, ICC-PCR (Integrated with Cell Culture PCR) and its modifications – have been used successfully for rapid detection of infectious enteroviruses and adenoviruses in environmental samples. The ICC-PCR technique is based on PCR analysis of the cell culture after incubation with test samples. Cell culture incubation allows elimination of non-infectious viruses and multiplication of infectious viruses, which then are detected by PCR. This method provides rapid and reliable detection of virus viability. Although ICC-PCR was developed originally for the detection of infectious viruses in environmental samples, the general principle of this method is applicable to detection of a replicating viruses in cells – for instance, the strand-specific PCR detection of Hepatitis C virus (HCV) in peripheral blood mononuclear cells. In this application, viral replication in cells is detected in terms of a HCV replicative intermediate, which is a negative-strand RNA. Detection on the basis of the negative strand means that false positive results due to amplification of RNAs of non-replicating viruses are avoided. Generally ICC-PCR is a very promising approach which with some modifications can be applied to identification of virus inactivation.

Volha Dzyakanava, PhD – Manager Virology Laboratory

To be successful in the marketplace eye care cosmetics must be non-irritating to the consumer.  Traditionally, a Draize rabbit eye test has been used to determine the safety of cosmetic products before sale to the general public.  The Draize Test involves applying the products directly to an animal’s eye.  The animals are observed for up to 14 days for signs of irritation (redness, swelling, discharge, cloudiness, or blindness in the tested eye).  Animal rights concerns, the cost of testing, and current European legislation banning cosmetics that have been tested using animals begs for an alternative model.  The MatTek  EpiOcular™ Tissue Model is a highly reproducible human cell-based in vitro tissue model that can be used to replace the traditional Draize ocular irritation testing.  The EpiOcular™ tissue model consists of normal, human-derived cells similar to cells found in the cornea (eye).  These cells are cultured on specially prepared cell culture forms creating multi-layered structures that are mitotically and metabolically active and induce the same inflammatory response as the human eye and can be used for detection of ocular irritation.  

We performed a collaborative study with MatTek to determine if the EpiOcular™ tissue model could also be used to differentiate between ultra-mild eye care cosmetic formulations.  Ultra-mild classifications can not be determined using the standard Draize rabbit eye test due to the insensitivity to the low levels of irritation induced by these products.  For the mildness testing, 10 commercially available mascara products, all with non-irritating or hypoallergenic claims, were purchased representing a broad range of manufacturers including Almay, Revlon, L’Oreal, Maybelline, and Covergirl.  The mascara products were tested using the EpiOcular™ model with an extended time exposure protocol (up to 24 hours).  The mascaras were applied to the surface of the ocular cells and remained in direct contact with the tissues for 8, 16, or 24 hours.  Following the exposure times, the mascaras were removed and the tissues and exposed to MTT for 3 hours.  Following the MTT exposure the tissues were removed from the MTT and rinsed with a Phosphate Buffered Saline solution. An alcohol extractant was added to the tissues following the MTT exposure.  The extraction process was allowed to proceed overnight.  The optical density (color change) of the extract was read the following day using a spectrophotometer.  If the mascara was non-irritating to the cells, the cells remained viable and were able to metabolize the MTT, thereby reducing the MTT and creating a visible color change from red/orange to blue/purple.  The absence of this darkening indicated that the mascara was irritating to the cells.  These optical density readings were used to determine the percent (%) viability of each tissue following exposure to each of the 10 mascara products.  An ET-50 (the time it takes for the viability of the tissues to decrease to 50%) irritation score was then assigned to each mascara.   The commercially available mascaras showed ET-50 scores ranging from 8.7 hours to > 20 hours.  These results relate to very mild products whose irritation potential would not have been detected with the standard animal model.  As such, the extended time exposure protocol appeared to be a facile, cost-effective means to screen ultra-mild eye care cosmetics. 

LIV GRAVING AND JESSICA MCDONNELL

MRSA is a resistant variation of the common bacterium Staphylococcus aureus. It has evolved an ability to survive treatment with most antibiotics. Methicillin-resistant Staphylococcus aureus (MRSA) is a bacterium responsible for difficult-to-treat infections in humans.

The organism is often sub-categorized as Community-Associated MRSA (CA-MRSA) or Health Care-Associated MRSA (HA-MRSA) although this distinction is complex. Some have defined CA-MRSA by criteria related to patients suffering from an MRSA infection while other authors have defined CA-MRSA by genetic characteristics of the bacteria themselves. The new CA-MRSA strains have rapidly spread in the United States to become the most common cause of cultured skin infections among individuals seeking medical care for these infections at emergency rooms in cities. These strains also commonly cause skin infections in athletes, jail and prison detainees, and soldiers.

Solution:
The overwhelming response in regards to the management and reduction in infection rates of these two pathogenic organisms were very basic; wash your hands regularly with an antimicrobial product and disinfect surfaces with a hard surface disinfectant–simple solutions to a very big problem.

BioScience Laboratories, Inc. has the capability of testing antimicrobial products, both topically applied antiseptics and hard surface disinfectants, for their efficacy against these organisms. To determine if a product is effective in killing these bacteria, standard in-vitro evaluations can be used to determine their efficacy. To take it a step further, one can evaluate their products’ efficacy in an IN-USE setting; that is, surrogate clinical trials on human volunteers. BSLI has recently been approved to conduct efficacy evaluations against C. diff and MRSA on Human Subjects. By working with an Institutional Review Board, BSLI has developed a Protocol to test the efficacy of topical antiseptic products on the skin. We feel that this type of testing will aid in the development of new novel antimicrobial products that can be used in day to day life as well as in hospital settings to help protect lives and reduce infections rates worldwide.

Kyle McGovern
Senior Account Executive