Confessions of a Cookie Mom

I decided last year that I would be a troop leader for my daughter’s Girl Scout Daisy troop. I thought it would be a great way to spend some one-on-one time with her, while volunteering for a cause I believe in. It started off positive; I met with the other leader, we had all these great ideas for what we would do! This was going to be so fun! Well, time went on, and the meetings seemed longer and longer;  the craft ideas were getting much less creative, and by the end of the year the girls were playing tag for the last 15 minutes of the meetings because we had run out of things to do. I realized that I really am not the best “traditional” troop leader, but I still wanted to be involved in a leader capactiy. I know! I will be the cookie coordinator for the Girl Scout cookie season! I can use the skills I am much better at (organization, metrics), and not have to be creative or crafty. Perfect!

Maybe I could arrange them into a futon frame or bookshelf?
Maybe I could arrange them into a futon frame or bookshelf?

We are two weeks into cookie season, and I would like to give the job back. I am completely obsessed with cookies. I think about them almost all day; I dream about them at night. I am placing cookie orders, counting cookies, schlepping cookies, EATING cookies. Here is a picture of my family room. This is what it looks like all the time. I have a “cookie spreadsheet”. I have nightmares that it will get accidentally deleted somehow, and that would throw my life into a tailspin. Continue reading “Confessions of a Cookie Mom”

Ghost of the Himalayas (Snow Leopard) Caught on Camera In a Successful Hunt

Snow LeopardPaul Steyn has posted an amazing series of photographs taken by Adam Riley in Hemis National Park in the Himalayas on the National Geographic News web site. These photographs are the  first photo documentation of a successful snow leopard hunt, and underscore the amazing biology of this area. If you are interested in learning more, we have another blog post about early work to isolate induced pluripotent stem cells from adult animals–an attempt to compliment the habitat preservation and other efforts to save this incredible hunter before it truly becomes a ghost.

Butterfly Heal Thyself: What We Can Learn from Self-Medicating Animals in Nature

Can animals actively seek out a plant to heal a parasitic infection or to numb pain? For years scientists have presumed that such activity was restricted to animals capable of more complex cognitive abilities, like chimpanzees. However scientists have described incidents of fruit flies (1), caterpillars (2)  and butterflies  (3) “self medicating” to prevent or cure parasitic infections, suggesting that such behavior may be both innate as well as learned.

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Fun with Chemistry for Valentine’s Day

Image credit: Morguefile

Looking for last-minute gift ideas? Wondering what chemistry has to do with Valentine’s Day? The chemists of the American Chemical Society (ACS) have curated a web page called Valentine’s Day Chemistry that offers gift ideas (for example, make a crystal heart using pipe cleaners, borax and hot water), explains the chemistry behind chocolate and flowers, shows a video that equates chemical bonding to people interacting at a party and more. The links and videos offer something for everyone, whether you want to have fun with friends and family of all ages or just want to learn something new about chemistry.

Happy Valentine’s Day!

Dealing with PCR Inhibitors

Inhibition

The polymerase chain reaction (PCR) has revolutionized modern biology as a quick and easy way to generate amazing amounts of genomic data. However, when PCR doesn’t work, it can be frustrating. At these times, PCR and reverse transcription PCR (RT-PCR) inhibitors seem to be everywhere: They lie dormant in your starting material and can co-purify with the template of interest, and they can be introduced during sample handling or reaction setup. The effects of these inhibitors can range from partial inhibition and underestimation of the target nucleic acid amount to complete amplification failure. What is a scientist to do?

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For Alphavirus Reporters, Location Matters

Computer-generated model of an alphavirus.
Computer-generated model of an alphavirus.

Luminescent reporters offer virologists a convenient way to measure replication of viruses and are also used to image the spread of viruses in vivo in experimental systems. These reporter viruses are useful for evaluating the effects of antiviral drug treatments, testing the efficacy of potential vaccines, and studying the ways in which viruses replicate in the body and cause disease. One challenge in the construction of such reporters is the need to ensure that the reporter molecule itself does not alter the virus in ways that affect its ability to cause disease. Another challenge is maintaining the reporter gene throughout several cycles of viral replication. In smaller viruses, it can be particularly difficult to introduce a reporter gene without compromising the ability of the virus to replicate and cause disease.

A 2014 paper was published in J. Virology comparing the effectiveness of various NanoLuc® luciferase alphavirus reporter constructs. The authors of the study, Chengqun Sun et al. from the University of Pittsburgh, placed these reporter genes in three different locations in the genome of several alphaviruses and compared the effect on their ability to replicate in vitro and in vivo. They also assessed the ability of the luciferase genes to persist during infection of cultured cells and in a mouse model. They showed that the size and location of the reporter had a significant effect on successful replication and persistence. They also showed that the reporters could potentially be integrated at different positions within the genome to study different aspects of viral pathogenesis.

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Battling Obesity One Thermostat at a Time

ThermometerWinter in Wisconsin is synonymous with cold, and this year thanks to the “wobbly polar vortex” it has been really, really cold. I have been very grateful for my under-desk space heater at work and my toasty gas fireplace at home. However, according to an article currently in press in the journal Trends in Endocrinology and Metabolism (1), all that lovely toasty warmness might be working against me if I am hoping to loose weight.

Excess weight is nothing more than a positive energy balance, meaning we have taken in more calories than we have burned. The deceptively simple sounding solution for losing or maintaining weight is to take in no more calories than you will be expending. Typically this is achieved by eating less, increasing physical activity or through pharmacological interventions. However, anyone who has ever tried any of these approaches knows that there is nothing simple about them, and often times the results are disappointing or short lived.

The authors of the Trends in Endocrinology and Metabolism paper hypothesize that regular exposure to mild cold could affect our energy expenditure in a positive way. We know that shivering produces heat (shivering thermogenesis) and thus expends energy. It can increase the metabolic rate to up to five  times the resting rate (2). However, it is not comfortable and can make coordinated movements difficult. The authors focused on something close to this state, but not quite. Nonshivering thermogenesis (NST) is the cold-induced increase in heat production that is does not involve muscle activity like shivering. It occurs by activating brown adipose tissue (3–5).

Although there is great variation between individuals, most young and middle-aged people will see an increase in NST between a few percent to up to 30% when exposed to mildly cold conditions (1). The authors note that a recent study found a significant decrease in body fat content following a 6 week cold acclimation study (2 hours/day at 62.6°F [17°C]; 6).

ScaleAs many of us can probably attest, the downfall of undertaking anything that increases our body’s energy usage is that often we end up increasing our caloric intake to compensate for it. Interestingly, the authors point to a 2009 study (7) that found evidence that increased food intake did not fully compensate for this type of cold-induced thermogenesis.

Personally, I love to be warm— as evidenced by my previous comments about space heaters and gas fireplaces. However, if the benefits of cold acclimation suggested by this paper hold to be true, I think I could tolerate 2 hours a day at 62°C.

References

  1. Van Marken Lichtenbelt. W. et al. (2014) Cold exposure—An approach to increasing energy expenditure in humans. Trends Endrochron. Met. In Press.
  2. Jansky, L. (1998) Shivering. In Physiology and Pathophysiology of Temperature Regulation (Blattheis, C.M. ed.) World Sceintific.
  3. Cannon, B. and Nedergaard, J. (2004) Brown adipose tissue: Function and physiological significance. Physiol. Rev. 84, 277–359.
  4. Van Marken Lichtenbelt, W.D. et al. (2009) Cold-activated brown adipose tissue in health adult men. N. Engl. J. Med. 360, 1500–1508.
  5. Virtanen, K.A. et al. (2009) Functional brown adipose tissue in health adults. N. Engl. J. Med. 360, 1518–1525.
  6. Yoneshiro, T.  et al. (2013) Recruited brown adipose tissue as an antiobesity agent in humans. J. Clin. Invest. 123, 3404–3408.
  7. Cannon, B. and Nedergaard, J. (2009) Thermogenesis challenges the adipostat hypothesis for body-weight control. Proc. Nutr. Soc. 68, 401–407.

Practical Tips for HEK293 Cell Culture When Using cAMP-Glo™ Assay

HEK293 cells stably expressing HaloTag®-ECS (ExtraCellular Surface; comprised of a signal sequence and single transmembrane domain of β1-integrin) fusion protein labeled with HaloTag® Alexa Fluor® 488 Ligand and then imaged.
HEK293 cells stably expressing HaloTag®-ECS fusion protein labeled
with HaloTag® Alexa Fluor® 488 Ligand and then imaged.

G Protein Coupled Receptors represent one of the largest classes of cell surface receptors and one of the most important classes for drug targets. Fifty of the top 200 drugs target GPCRs. GPCRs respond to various stimuli like light, odors, hormones, neurotransmitters and others. They cover virtually all therapeutic areas. When a particular GPCR is implicated in a disease, researchers screen the GPCR and its signaling pathways, the hope being that promising therapeutic targets might be identified. Major G-protein families signal via secondary messengers like cAMP, which in turn activate a range of effector systems to change cell behavior and/or gene transcription. There are various approaches and methods to study GPCRs and measure the increase or decrease of intracellular cAMP. However, the fastest and the most sensitive among all methods is a plate based cAMP-Glo™ Assay.

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Using NanoLuc® Luciferase to Study Interactions between Environmental Flavobacteria and Mosquitos

nanoluc

Studies of the larval stages of Aedes triseriatus  (Eastern TreeHole Mosquito) indicate that the “tree hole” habitats in which these larva develop contain diverse microflora including the flavobacteria Elizabethkingia and Chryseobacterium. Extracts from these bacteria have many properties that might affect mosquito health, including antibacterial and anti-fungal activities. Understanding how these bacteria affect larval mosquito development has the potential to inform strategies for mosquito control.

Some initial work has been done by expressing Bacillus larvacidal toxins in some species of Gram-negative bacteria. However, only limited success was achieved using laboratory bacterial strains for such studies. Using environmental flavobacteria might prove to be a more useful approach. However, few molecular tools exist to study environmental flavobacteria. GFP reporters have been used to look at larval feeding, but autofluorescence in the pupae limit the usefulness of GFP-labeled strains for quantitative studies. Furthermore, environmental flavobacteria have unique transcription and translation machinery, and selectable markers and expression plasmids from proteobacteria do not function in these wild strains.

Chen and colleagues set out to generate molecular tools to study Flavobacterium hibernum, a fast-growing bacterium from native mosquito habitats. Their goal was to use these tools to see if A. triseratus larvae ingest and digest these bacteria and to test whether or not F. hibernum can be used to as a vector for larvacidal toxins directed against mosquito larvae. The results of their work were published in Applied and Environmental  Microbiology .

To develop a reporter that avoided issues of autofluorescence background for quantitative studies on the feeding behavior, the researchers turned to NanoLuc® Luciferase, a small, bright luciferase derived from the sea shrimp Oplopphorus gracilirostris. This luciferase has been used in mammalian cells for many kinds of studies, but it has not been used as a reporter in bacterial cells prior to the work of Chen et al. They also looked at work with laboratory flavobacteria strains that used a promoter of outer membrane protein A (PompA) to drive reporter expression as a potential system that might also work with environmental flavobacteria strains.

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Amino Acid Analogs as Possible Cancer Drugs

HeLa cells stained with Hoechst 33258. By TenOfAllTrades (From English wiki 1.) [Public domain], via Wikimedia Commons
Scientists look in unusual places for potential anticancer treatments. I have reviewed papers that investigated the possibility that dandelion root may harbor anticancer treatments, milk fat may moderate cancer metastasis and the effects of chemotherapy, and black raspberry extract may even prevent cancer. Sometimes, research avenues come down to an observation about what a tumor cell needs to grow and exploring the idea that molecular analogs might be a tool to block cancer growth. For the work reported in Drug Design, Development and Therapy, analogs of the amino acid glycine, specifically glyphosate and aminomethylphosphonic acid (AMPA), the degradation product of glyphosate, were used to explore this idea in cancer cell lines. Continue reading “Amino Acid Analogs as Possible Cancer Drugs”