Friday, March 29, 2013

Focus on: yesterday's webinar

It was truly a pleasure for BMG to be able to present with out colleagues from Teacan, Douglas Scientific and BioTek during yesterdays webinar. Everyone did a great job and thank you to all of you that were able to attend and ask such great questions!

If you were unable to attend please check the webinar page on the BMG LABTECH website to see when it becomes available to view.

Thursday, March 28, 2013

Applications Thursday: High-Throughput MEKK2 Intrinsic ATPase Activity Assay

In the current issue of the Journal of Biomolecular Screening the Scott lab from North Carolina Central published a paper entitled: 'Development and Validation of a High-Throughput Intrinsic ATPase Activity Assay for the Discovery of MEKK2 Inhibitors'. The kinase MEKK2 is implicated in cancer so a selective inhibitor could be a useful therapeutic agent. In the current report the authors exploit the finding that MEKK2 can hydrolyze ATP even when there is no substrate for MEKK2 to phosphorylate.

Following incubation of MEKK2 with ATP the production of ADP could be monitored using the ADP-Glo Reagent from Promega and the resulting luminescence was detected using the PHERAstar microplate reader from BMG LABTECH. Using the PHERAstar they were able to perform detection in a high-throughput assay that had average Z' values of 0.72; indicating that this assay will be reliable for high-throughput screening. An initial screen for inhibitors of MEKK2 ATPase activity using this high-throughput format revealed 2 novel inhibitors.



To find out more about the diverse applications that the PHERAstar and other BMG LABTECH microplate readers can perform please visit the applications center on the BMG LABTECH website.

Wednesday, March 27, 2013

Webinar tomorrow: Improvements in Microplate Technology and Its Impact on Your Efficiency

Tomorrow (March Thursday March 28) starting at 12:00 p.m. Eastern Time BMG LABTECH is co-sponsoring a webinar. During this webinar, which is free to laboratory professionals, the increasing importance of microplate readers and associated technologies will be discussed as they pertain to a variety of endeavors including; drug discovery and assay validation.

BMG LABTECH is The Microplate Reader Company and we pride ourselves on our ability to consistently raise the bar for microplate reader performance. To this end, during this webinar, BMG will discuss its latest technology. A microplate reader that will allow you to perform ANY assay since it can perform detection at ANY wavelength and ANY bandwidth! These capabilities are the result of this being a high performance advanced monochromatic microplate reader with a spectrometer and filters.

We hope that you will join us!



http://www.labmanager.com/?articles.view/articleNo/4900/title/Webinar--Improvements-in-Microplate-Technology-and-Its-Impact-on-Your-Efficiency/


Tuesday, March 26, 2013

BMG LABTECH: Improving Technology Enhancing Your Efficiency


On Thursday March 28, at 12:00 p.m. - 1:30 p.m. Eastern Time; BMG LABTECH will participate in the free webinar entitled: Improvements in Microplate Technology and Its Impact on Your Efficiency. During this webinar BMG will discuss a new microplate reader! This reader builds on the filter based technology and  spectrometer that have made our previous microplate readers so useful. The improvement in technology for this reader comes from the advanced monochromator! 

To find out how this new high performance microplate reader will allow you to measure ANY wavelength and ANY bandpass thus allowing you to perform ANY assay attend Thursdays webinar!

FAQ: What are xenonucleic acids?

It has long been hypothesized that due to their complexity neither DNA nor RNA could have arisen spontaneously. Because of its simpler structure and ability to perform other functions beyond information storage it has been proposed that RNA preceded DNA. However, it is still too complex to have been formed in the prebiotic world. Therefore a variety xenonucleic acids are being studied as possible candidates for the transitional nucleic acid that could be a precursor to RNA and DNA. A recent study published in the Journal of the American Chemical Society has investigated threose nucleic acid (TNA).

Similar to DNA and RNA, TNA is composed of repeating nucleotide bases held together by a sugar phosphate backbone. However for each the sugar is distinct; being either deoxyribose, ribose or threose. TNA is also similar to DNA in that it can form a double helix but employs a 4 rather than 5 carbon sugar which could arise from two 2 carbon precursors. Despite this distinction TNA can still bind with complimentary DNA and RNA strands and therefore act as an alternative self-replicating entity.

The research group from Arizona State University used commercially available synthetic polymerases to transcribe DNA to TNA and reverse transcribe TNA to DNA with 90% accuracy. These results should allow for additional analysis of other xenonucleic acids and may indicate that TNA paved the way for RNA and subsequently DNA and the emergence of earthly life.

Monday, March 25, 2013

Did you know: DNA damage could be a part of normal learning?

 DNA ligase encircles the double helix
to repair a broken strand of DNA
According to a recent report in Nature Neuroscience a form of DNA damage called double strand breaks (DSB's) exhibit an increase in mice brains that are exposed to new environments. DSB's have long been thought to be associated with age-related illnesses such as Alzheimers. However, this report shows that they are formed during normal brain function as well and may be important in the conversion of neuronal DNA into proteins that are involved in forming memories.

The collaborators from San Francisco were comparing normal mice to genetically modified mice that mimic Alzheimers and were initially surprised to find the DSB's in normal mice. However, after an initial rise in DSB's the levels began to decrease suggesting that a cycle of damage and repair is normal in the learning process. In contrast, the modified mice had higher initial levels of DSB's which rose when exposed to a new environment but were decreased more slowly. So while some formation of DSB's is normal, if the levels are not decreased permanent damage could result.

Article citation:

Physiologic brain activity causes DNA double-strand breaks in neurons, with exacerbation by amyloid-β

Friday, March 22, 2013

Focus on: Events

BMG LABTECH will be attending both JSBBA 2013 and GEN2BIO next week!


The Japan Society for Bioscience, Biotechnology and Agrochemistry annual meeting is being held March 25th to 27th  and BMG LABTECH will be there!



BMG representatives will also attend GEN2BIO, the biotech congress organized by Biogenouest, on March 28th in France.

Thursday, March 21, 2013

Applications Thursday: ERK 1/2 phosphorylation using AlphaScreen SureFire

Activation of of ERK1/2 is central to a number of signaling pathways that convert extra-cellular signals that bind to receptors into cellular responses such as changes in transcription/translation. As a result, monitoring the phosphorylation status of ERK1/2 as an indication of activation can be used as a read out of receptor function and is therefore a useful tool in screening of compounds that alter receptor function.

AlphaScreen measurement of activated
Gi and Gq coupled GPGR's
Analysis of ERK phosphorylation can be done in a high-throughput format using the AlphaScreen  SureFire Phopho-ERK1/2 assay which is based, in principle, on an immuno-sandwich assay. The result is that when ERK1/2 is phosphorylated a donor bead and an acceptor bead are brought into close proximity. Laser excitation of the donor converts ambient oxygen to singlet oxygen initiating a cascade of chemical reactions that result in a luminescent signal in the acceptor bead.

In application note 210 from BMG LABTECH this approach was used to produce dose response curves for Gi and Gq G-protein coupled receptor agonists in a 384 well format using the POLARstar Omega. Although the POLARstar Omega does not have a laser for excitation, the use of AlphaScreen optimized filters allow this microplate reader to consistently attain sensitive results.

For more information on this and other applications for all of the microplate readers that BMG LABTECH has to offer please visit the applications center on our web site.

Wednesday, March 20, 2013

Fun fact: Neuron firing captured in zebrafish

Coronal section containing the chronically imaged pyramidal neuron “dow”
doi:10.1371/journal.pbio.0040029
A report in Nature describes the first imaging of a vertebrate brain at the single cell level. The report describes how zebrafish larvae were genetically engineered such that they produce a protein that glows as a result of chemical changes from neuron firing. This modified fish and a the use of high-speed light-sheet microscopy allowed imaging of more than 80% of the approximately 100,000 neurons present in the larval brain every 1.3 seconds. This type of whole brain activity mapping could eventually be used to help explain brain function.

Tuesday, March 19, 2013

FAQ: What is the human gene connectome?

As a result of high through-put genome sequencing technologies a plethora of data can be generated from patient samples. However, this data can take a lot of time to search through in order to find the relevant disease causing mutation in a monogenic disease hidden within this data. In hopes of making this search more directed, a collaboration of scientist from the US, France and Israel sought to design a computer program that would determine the biological distance between every gene in the human genome. The report on the human gene connectome (HGC) appears in PNAS.

Human_genome.png
The HGC is based on what has been discovered about the protein-protein interactions and molecular pathways in which gene products exist and applies a numerical value to how closely connected two genes are. As support of principal the group applied this approach to a disease. They studied the pathogenesis of herpes simplex virus 1 (HSV-1) encephalitis (HSE). Inborn errors of Toll-like receptor 3 (TLR-3) have been shown to be important in HSE pathogenesis, so they developed a TLR-3 specific connectome. The results confirmed what many had previously shown regarding other genes in the molecular pathway which when mutated are also associated with the disease state. In particular, when genetic data from two patients was compared they were both confirmed to have a mutation in TANK-binding kinase which is a biologically close gene and has previously been associated with HSE.

The authors believe that using the HGC will significantly increase the discovery of disease-causing genes by solving the needle in a haystack problems associated with extensive genetic data sets. The HGC program is freely available to noncommercial users:  http://lab.rockefeller.edu/casanova/HGC

Article: The human gene connectome as a map of short cuts for morbid allele discovery





Monday, March 18, 2013

Did you know: some species of bacteria produce toxins that can kill themselves?

Mattes of cyanobacteria. Looking like an image from space,
 mattes of cyanobacteria thrive in the scalding waters of Biscuit Basin,
Yellowstone National Park, Wyoming. USA, Yellowstone National 
Cyanobacteria are more commonly known as blue-green algae, some of which produce cyanotoxins that can be harmful to themselves and other species. This production of toxins is part of a system in the cyanobacterium Synechocystis that assure that the plasmid pSYSA is maintained in their lineage. Research performed at the University of Freiburg and published in two recent articles propose that this is due to this plasmid containing genetic information for a bacterial immune system. The presence of this plasmid is maintained by seven toxin-antitoxin pairs on the plasmid.

The basis behind a toxin-antitoxin system is that as long as the plasmid is present both toxin and anti-toxin are continually produced and the bacteria survives. However, the toxin that is produced is much more stable than the anti-toxin. So, if a bacteria loses the plasmid during cell division they lose the ability to produce anti-toxin but they are still in an environment that contains the stable toxin and therefore they perish.

There are a number of ways to monitor bacterial growth. But if you want to employ nepholometry in a microplate format only the NEPHELOstar Plus from BMG LABTECH allows you to do so.

Articles:

CRISPR-Cas Systems in the Cyanobacterium Synechocystis sp. PCC6803 Exhibit Distinct Processing Pathways Involving at Least Two Cas6 and a Cmr2 Protein



Toxin-Antitoxin Systems on the Large Defense Plasmid pSYSA of Synechocystis sp. PCC 6803

Friday, March 15, 2013

Focus on: Events

BMG LABTECH will be very busy in the next week attending conferences in Europe and Asia.

In the UK BMG representatives will be present for a meeting of the European Laboratory Interest Group. If you are in academia, biotech or industry, come see us at booth F1 to find out how we can assist in your early-phase drug discovery on March 19th.

If you are in Germany you can meet with our BMG team at the DisCovery Congress Chemistry on March 19th and 20th. Our representatives will be on hand at booth 8 talk about integrating the latest structural chemistry with innovative technologies for drug discovery.

On the the 20th and 21st BMG representation will be at ChemBio in Finland.

Finally in China on March 21-23 BMG LABTECH will be at the 6th Annual World Protein and Peptide Conference. The self proclaimed: 'leading meeting place in the world for protein/peptide sciences'.

As you can see BMG LABTECH is available worldwide. Come see why we are The Microplate Reader Company.

Thursday, March 14, 2013

Applications Thursday: Real-time Quaking Induced Conversion Assay

RT-QuIC sensitivity: analysis of dilutions of scrapie hamster
brain homegenate stock at indicated concentrations. The 50%
seeding dose was achieved with a 2 ul aliqout (seeding dose)
of a 10E-9 dilution of the stock (dark blue line)
From BMG application note 232
The RT-QuIC assay allows you to perform quantitative analysis of prion seeding activity using the FLUOstar Omega from BMG LABTECH. This approach has been applied to many types of prion assays including sheep scrapie and deer chronic wasting disease.

The recent publication: 'Prion Seeding Activities of Mouse Scrapie Strains with Divergent PrPSc Protease Sensitivities and Amyloid Plaque Content Using RT-QuIC and eQuIC' describes the extension of this application to mouse models. This is important as murine models are central to transmissible spongiform encephalopathy pathogenesis research.

Using this approach the authors were able characterize multiple scrapie strains and predict that there will be multiple interesting applications of this approach for mouse adapted strains.

For more information on this application please read application note 232 available on the BMG LABTECH website.


Wednesday, March 13, 2013

Alleles of mouse prion protein affect time to aggregate formation


Prion diseases are neurodegnerative disorders where a the normal cellular form of the prion protein polymerizes into an infectious form that induce formation of repeating sheets of protein, the effect of which is eventually fatal. The sequence of host prion protein is the main determinant in whether disease formation occurs.
PrPsc induces conformational change in PrPc
http://en.wikipedia.org/wiki/User:Joannamasel

A recent paper in the Journal of Biological Chemistry has studied two forms of mouse prion protein that are polymorphic at amino acid residues 108/189. The results have implications in understanding disease susceptibility and species barriers.

The group from University of Alberta found that the recombinant forms of the two prion protein differed with respect to aggregation and response to seeding in an in vitro kinetic assay.

Tuesday, March 12, 2013

FAQ: How does periconceptional nutrition affect a childs' health

Periconceptional nutrition or the food a that a mother consumes in the days and weeks around the time of conception has been linked to the health of the child. A recent paper in PLoS Genetics presents findings that indicate that differences in nutrition lead to epigenetic alterations, specifically an alteration in DNA methylation at regions of five genes.

The collaborative study funded by the US Department of Agriculture studied children born in the West African nation of Gambia. They found that children conceived when food is less available during the peak of the rainy season  in August and September exhibited an increase in DNA methylation.

Same Genes but a Different Kink in the Tail
While a mutation in the axin gene called axin-fused
 produces mice with kinky tails, the degree of kinkiness
varies among genetically identical littermates

http://dx.doi.org/10.1371/journal.pbio.0000082


The goal of the research was to identify metastable epialleles (MEs), which in mice have been shown to cause obvious  phenotypic variation among genetically identical mice. They provide evidence that MEs do exist in the human population and that DNA methylation at such sites is altered dependent on the maternal environment.

This is just the latest example of the emerging importance of epigenetics. At BMG LABTECH we understand this importance and we have applications that can be used in your epigenetics studies.

Article: Season of Conception in Rural Gambia Affects DNA Methylation at Putative Human Metastable Epialleles

Monday, March 11, 2013

Did you know telomere length is an indication of life expectancy?

Telomere in red, human chromosomes in blue
Reinhard Stindl
In a study presented at the American College of Cardiology's Annual Scientific Session investigators from the Intermountain Heart Institute shared results that indicate that telomere length can predict life expectancy. Telomeres are a region of repetitive sequence that occur on the ends of chromosomes. Their purpose is to protect the ends of the chromosome from damage and have been used as a measure of age due to the fact that telomeres get shorter with age until cell division ceases. This shortening is also correlated with an increase in age related diseases.

In the current study samples from more than 3500 stroke and heart attack patients were employed and showed that if the results are adjusted for age of patients, longer telomere length directly correlated with life expectancy. The study employed the very complete records kept and the extensive follow up data and samples collected by the Intermountain Heart Institute. This allowed                                                                                                                                     them to follow any changes in the length of a patients telomeres over time.

It is hoped that telomere length could be used to measure the effectiveness of treatments along with tests that are currently employed such as cholesterol and blood pressure.

Friday, March 8, 2013

Focus on: Events

Next week BMG LABTECH will be at the Queenstown Molecular Biology Meeting & The 5th National Forum on New Technologies in Drug Discovery. The meeting is on March 11-12 at the Chinese National Compound Library in Shanghai, China. Please stop by and meet with BMG representatives to find out how our technology can assist your drug discovery program.

For information on this and other events visit the BMG LABTECH events page.

Thursday, March 7, 2013

Applications Thursday: Fluorescence Polarization based High Throughput Screening for inhibitors of bacterial Lipopolysaccharide production

Bordetella_bronchiseptica.jpg
With the increasing concern over antibiotic resistant bacteria it becomes more important to identify other mechanisms that can assist in halting bacterial growth. A potential untapped target are the enzymes that produce components of the lipopolysaccharide (LPS) outer membrane of Gram-negative bacteria. A paper in the recent issue of the Journal of Biomolecular Screening has used a PHERAstar plate reader from BMG LABTECH to perform a novel HTS using a FP based assay  in an attempt to identify inhibitors of LpxA. This enzyme is involved in  the synthesis of the Lipid A component of LPS.

The paper entitled: 'A High-Throughput-Compatible Fluorescence Anisotropy-Based Assay for Competitive Inhibitors of Escherichia coli UDP-N-Acetylglucosamine Acytransferase (LpxA)' is the work of scientists from AstraZeneca in Boston, MA. Their approach reveals inhibitors of LpxA that are able to displace a fluorescent ligand from the active site of LpxA resulting in a decrease in FP signal.

Diagram of PHERAstar FP optic module
Key to the success of this application is the optic modules employed by the PHERAstar which allow for this application to employ low volumes and lowest possible LpxA concentration and still attain satisfactory performance.

Wednesday, March 6, 2013

CDC is concerned about bacteria that are resistant to strong antibiotics and you should be too

Carbapenem-resistant Enterobacteriaceae (CRE) are currently found only in hospitals and nursing homes but their prevalence is on the rise and there is concern among the health care community that their resistance could spread to other bacteria. CRE bacteria contribute to death in nearly 50% of patients who become infected and are resistant to almost all available antibiotics. Furthermore, the antibiotics to which they are not resistant are quite toxic to humans as well.

Schematic drawing of bacterial conjugation. Conjugation diagram
1- Donor cell produces pilus. 2- Pilus attaches to recipient cell,
brings the two cells together.3- The mobile plasmid is nicked
and a single strand of DNA is then transferred to the recipient cell.
 4- Both cells recircularize their plasmids, synthesize second strands,
and reproduce pili; both cells are now viable donors
It is possible that CRE will be able to pass along the resistance genes that it carries through conjugation; a common bacterial process that allows bacteria to transfer genetic material to other bacteria through cell to cell contact. If this happens common conditions which affect a larger portion of the population could become untreatable.



The CDC is enacting procedures at hospitals and long term care facilities around the country to contain the spread of CRE. But what can you do? First take antibiotics only when prescribed and request antibiotics only when absolutely necessary.



If you are a patient in a hospital or long-term care facility the CDC suggests that you:

1) Tell your doctors if you have been hospitalized elsewhere

2) Clean your hands often

3) Expect health care providers to do the same and to clean their hands before and after touching you or the tubes going into your body

4) Ask anyone who visits you to clean their hands before and after their visit

5) Understand what is being done to you. Ask questions!




Tuesday, March 5, 2013

FAQ: What is protein nitration?

Nitrated tyrosine
Nitration, in general, is the process of introduction of a nitro group (-NO2). For proteins it represents a post-translational modification that occurs primarily on tyrosine residues. The effect of which seems to vary greatly.

A recent study published in PNAS entitled: "Nitration of Hsp90 induces cell death" describes how nitration as a result of oxidative stress can turn a normally helpful protein into a toxic protein. This 90 kDa heat-shock protein is a part of the highly conserved family of proteins that are well characterized for their function as chaperones. So for Hsp 90 to be able to convert into a protein that induces cell death may, at first, seem illogical. However, since Hsp 90 plays a role in up to 200 cell functions, when it is shut down by tyrosine nitration, all of those cell functions
are adversely affected.

Hsp90.jpg
The presence of tyrosine nitrated Hsp 90 is seen in a number of different pathologies and it is hoped that finding a drug that protects against tyrosine nitration may provide protection for a variety of degenerative diseases.

One of the main antioxidant systems employed to keep oxidative stress at bay is the thioredoxin reduction pathway. BMG LABTECH application note #167 describes how thioredoxin activity can be studied in neurons and glial cells using the FLUOstar OPTIMA. For more information visit our applications center.

Monday, March 4, 2013

Did you know: BMG microplate readers integrate with Staubli robots

BMG LABTECH is The Microplate Reader Company. As such we strive to be able to interface with any robotic system that you choose to employ. This includes the Switzerland-based company Staubli Robotics whose TP80 Fast Picker can sort at 200 picks per minute for loads of less than 0.1 kg or 170 picks per minute with its maximum load of 1 kg.

BMG users have proven success with Staubli products. Including at the Burnham Institute where the PHERAstar FS is used for a variety of applications, such as; fluorescence polarization and time-resolved fluorescence.

Friday, March 1, 2013

Focus on: Events

Coat of arms of the Royal Society of Chemistry
On March 4-5 BMG LABTECH will be at Fragments 2013, the Fourth Royal Society of Chemistry-Biological & Medical Chemistry Sector Fragment-based Drug Discovery meeting in Oxfordshire, UK.

The program will focus on new methods and technological developments as well as some screening results and approaches tailored to specific enzymes.

Please stop by and meet with the local BMG representatives to find out how a microplate reader from BMG LABTECH can benefit your drug discovery screening.