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ECOViEWS: Advertising is not just for humans – Aiken Standard

Posted: February 11, 2017 at 5:45 pm

Back when a phone booth was where Clark Kent went to don his Superman costume, inside each booth hung a phone directory. At the back of the directory was an alphabetical list of advertisements collectively known as the Yellow Pages. Today, few people have access to printed Yellow Pages. But advertisements are ubiquitous, which means you can still play what I call the Yellow Pages Game.

The rules are simple. How many advertised services and products can you find that parallel activities in nature? In other words, what advertisements targeted for people mirror the actions of plants and animals? A few, such as ads for legal services, are uniquely human, but not many.

We need not look beyond ads for the home to find countless examples. Heating and cooling, home security, house construction, all these are represented in the natural world. Take, for example, air-conditioning. Definitely not restricted to humans. Honeybees are the consummate climate control experts, maintaining the hive at a near constant temperature year-round. During hot weather, selected worker bees place water droplets in the hive and wave their wings in unison. Fanning the hive in that way can lower the temperature several degrees. During winter, workers become heater bees, continually flexing their muscles isometrically to warm the hive.

Home security is essential throughout the plant and animal kingdoms. Honeybees, wasps and hornets are on constant guard against intruders and have the weapons to defend their homes. A plants home is the plant itself, and numerous plants produce toxins or thorns that ward off many of their predators. Sensitive mimosa plants have what is equivalent to an alarm system. When a stem is disturbed by a hungry insect, the leaves can rapidly fold up, which may startle the insect into leaving. Some acacia trees of Africa have an association with ants that live on them and protect the trees from plant-eating insects. The ants attack an invading beetle or aphid that plans to make a meal of acacia leaves. The ants, meanwhile, get their nutrients from the acacia tree, much like a homeowner might feed his Doberman. Either is a good security system.

Home construction is common to innumerable animals. Honeybees come to mind again, with their elaborate combs of hexagonal cells that serve as storage for honey and for developing larvae and pupae. The American beaver is not to be outdone in home building, with its dams and lodges, and most avian species build something humans would have difficulty duplicating bird nests. Prairie dog colonies consist of tunnels, passageways and side chambers the quintessential underground estate.

And what about advertising itself? Do plants and animals advertise? Absolutely. The phenomenon is all around us. As spring arrives, we will see myriad examples in flowers with their colorful displays designed to attract pollinating insects. Lots of fish and lizards are also more brightly colored during mating season. Males of many of the usually drab darters, small fish that have their greatest biodiversity in Alabama, are noted for their spectacular color displays during the breeding season. As with humans, sound is a common advertising tool. Frogs and birds give mating calls in an effort to attract mates. Fireflies, both males and females, use lights to advertise their availability to procreate.

As with our own commercials, false advertising must constantly be guarded against. The deep-sea angler fish produces a light that dangles in front of its mouth and means sure death for a would-be predator trying to grab a morsel in the dark. Pitcher plants use sweet-smelling aromas to lure unsuspecting insects into the trumpet-shaped tube filled with liquid. Bola spiders have their own scam, producing chemicals that mimic the perfume-like pheromones that female moths use to attract mates.

Advertising is a part of life worldwide, and most human merchandise and trades that are advertised are duplicated in nature. Finding analogies can be a fun and enlightening exercise at home or in the classroom.

Whit Gibbons is an ecologist and environmental educator with the University of Georgias Savannah River Ecology Laboratory. Send environmental questions to ecoviews@gmail.com.

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Why Bioethics Matters in Biotechnology – Azusa Pacific University

Posted: February 11, 2017 at 5:45 pm

The last five years have witnessed amazing acceleration of innovation in biotechnology. CRISPR will lead to precision gene editing that could vastly improve food crop yields and provide cures to cancer. Lightning-fast gene sequencing will enable early detection of cancer from a simple blood test. High-speed bulk data transfer allows the entire genomes of millions of people to be compared online in the search for cures to both common and rare diseases. Neuromorphic chips will accelerate the dawn of artificial intelligence, and smart prostheses will allow para- and quadriplegic patients to move, the deaf to hear, and the blind to see.

Discovery of synergies in applications that blur the boundaries of traditional science, technology, engineering, and mathematics will continue to fuel this exponential growth of innovation. In spite of this exuberant trend, it is important to remember that innovation and discovery often outpace the regulatory structures that ensure their best and most ethical use in society.

The bioethics field traditionally is interpreted as pertaining mainly to the medical interests of humans. It has dealt with five key issues: beneficence, non-maleficence, patient autonomy, social justice, and patient confidentiality. However, with the advent of nanotechnology and other technologies that allow inter-kingdom transfer of genetic material, a need exists to establish a broader interpretation. Theologian Brian Edgar1 notes that a more robust definition should comprise six key considerations: respect for the intrinsic value of all life, valuing human uniqueness, preserving organismal integrity, recognizing ecologic holism, minimizing future liability, and producing social benefit. These considerations, while not expected to provide all of the answers to ethical dilemmas faced by technological advancement, create a framework for productive discussion of the most important aspects of biotechnology.

As Christians, we must also acknowledge that we are made in the image of God2, and have the unique ability, of all created things, to have a relationship with our Creator. In thoughtfully considering the implications of having been thus created, we have the responsibility of honoring Him by not only valuing human life, but by valuing and caring for His creation as well. If we actively and consistently apply this principle to guide us in making decisions about the application of biotechnology, the benefits to ourselves and to our world will be tremendous.

Posted: February 10, 2017

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OCASCR scientists make progress in TSET-funded adult stem cell research – NewsOK.com

Posted: February 11, 2017 at 5:44 pm

OCASCR scientist Lin Liu at work. Photo provided.

Working together, scientists from Oklahoma State University, the University of Oklahoma Health Sciences Center and the Oklahoma Medical Research Foundation are advancing adult stem cell research to treat some of todays most devastating diseases.

Under the umbrella of the Oklahoma Center for Adult Stem Cell Research (OCASCR), created with funding from the Oklahoma Tobacco Settlement Endowment Trust, these scientists have amassed groundbreaking findings in one of the fastest growing areas of medical research.

We have made exciting progress, said OCASCR scientist Lin Liu, director of the Oklahoma Center for Respiratory and Infectious Diseases and director of the Interdisciplinary Program in Regenerative Medicine at Oklahoma State University.

We can convert adult stem cells into lung cells using our engineering process in petri dishes, which offers the possibility to repair damaged lung tissues in lung diseases, said Liu, whose research primarily focuses on lung and respiratory biology and diseases.

Using our engineered cells, we can also reverse some pathological features. These studies give us hope for an eventual application of these cells in humans.

Adult stem cells in the body are capable of renewing themselves and becoming various types of cells.

Until recently, stem cell treatments were largely restricted to blood diseases. However, new studies suggest many other types of adult stem cells can be used for medical treatment, and the Oklahoma Center for Adult Stem Cell Research was created to promote this branch of research.

OCASCR scientist Lin Liu and his team discussing their work. Photo provided.

Liu said the discipline provides hope for many ailments.

What most fascinated me in stem cell research is the hope that we may be able to use stem cells from our own body; for example, bone marrow or fat tissues to cure lung diseases, Liu said.

It is impossible to know exactly which diseases will respond to treatments.However, results of early experiments suggest many diseases should benefit from this type of research, including lung, heart, Alzheimers and Parkinsons diseases, as well as cancer, diabetes and spinal cord injuries. The field is often referred to as regenerative medicine, because of the potential to create good cells in place of bad ones.

While the application of stem cells can be broad, Liu hopes that his TSET-funded work will help develop treatments for diseases caused by tobacco use.

The goal of my research team is to find cures for lung diseases, Liu said. One such disease is chronic obstructive pulmonary disease (COPD).

COPD is the third leading cause of death in the country and cigarette smoking is the leading cause of COPD.

Cigarette smoking is also a risk factor for another fatal lung disease, idiopathic pulmonary fibrosis (IPF), which has a mean life expectancy of 3 to 5 years after diagnosis, he added.

There is no cure for COPD or IPF. The current treatments of COPD and IPF only reduce symptoms or slow the disease progression.

Using OCASCR/TSET funding, my team is researching the possibility to engineer adult stem cells using small RNA molecules existing in the body to cure COPD, IPF and other lung diseases such as pneumonia caused by flu, Liu said.

This is vital research, considering that more than11 million peoplehave been diagnosed with COPD, but millions more may have the disease without even knowing it, according to the American Lung Association.

Despite declining smoking rates and increased smokefree environments, tobacco use continues to cause widespread health challenges and scientists will continue working to develop treatments to deal with the consequences of smoking.

We need to educate the public more regarding the harms of cigarette smoking, Liu said. My research may offer future medicines for lung diseases caused by cigarette smoking.

Under the umbrella of the Oklahoma Center for Adult Stem Cell Research (OCASCR), created with funding from the Oklahoma Tobacco Settlement Endowment Trust, these scientists have amassed groundbreaking findings in one of the fastest growing areas of medical research. Photo provided.

Liu has been conducting research in the field of lung biology and diseases for more than two decades.

However, his interests in adult stem cell therapy began in 2010 when OCASCR was established through a grant with TSET, which provided funding to Oklahoma researchers for stem cell research.

I probably would have never gotten my feet into stem cell research without OCASCR funding support, he said. OCASCR funding also facilitated the establishment of the Interdisciplinary Program in Regenerative Medicine at OSU.

These days, Liu finds himself fully immersed in the exciting world of adult stem cell research and collaborating with some of Oklahomas best scientific minds.

Dr. Liu and his colleagues are really thriving. It was clear seven years ago that regenerative medicine was a hot topic and we already had excellent scientists in the Oklahoma, said Dr. Paul Kincade, founding scientific director of OCASCR. All they needed was some resources to re-direct and support their efforts. OSU investigators are using instruments and research grants supplied by OCASCR to compete with groups worldwide. TSET can point to their achievements with pride.

The Oklahoma Center for Adult Stem Cell Research represents collaboration between scientists all across the state, aiming to promote studies by Oklahoma scientists who are working with stem cells present in adult tissues.

The center opened in 2010 and has enhanced adult stem cell research by providing grant funding for researchers, encouraging recruitment of scientists and providing education to the people of Oklahoma.

We are fortunate that the collaboration at the Oklahoma Center for Adult Stem Cell Research is yielding such positive results, said John Woods, TSET executive director. This research is leading to ground breaking discoveries and attracting new researchers to the field. TSET is proud to fund that investments for Oklahomans.

Funding research is a major focus for TSET and it comes with benefits reaching beyond the lab. For every $1 TSET has invested at OCASCR, scientists have been able to attract an additional $4 for research at Oklahoma institutions, TSET officials said.

TSET also supports medical research conducted by the Stephenson Cancer Center and the Oklahoma Tobacco Research Center.

For more information, visit http://www.ocascr.org.

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Inhibition of the immunoproteasome may improve graft acceptance [PreClinical] – 2 Minute Medicine

Posted: February 11, 2017 at 5:44 pm

1. A targeted immunoproteasome inhibitor, DPLG3, was found to have high selectivity for the immune cell proteasome and was not toxic to other cell types.

2. When administered after a heart transplant in mice, DPLG3 allowed for minimal graft rejection through a decrease in T cell activation.

Evidence Rating Level: 1 (Excellent)

Study Rundown: Organ transplants typically require the recipient to continue taking immunosuppressive medications to prevent rejection of the graft. However, the toxicity of these medications necessitates a safer option for transplant recipients. Researchers in this study synthesized DPLG3, a molecule that was modeled after a currently prescribed proteasome inhibitor and had selectivity to a subunit of the immunoproteasome.

When tested in vitro, DPLG3 selectively inhibited the 5i immunoproteasome subunit in a lymphoma cell line and did not affect other immunoproteasomes or cell types. The compound was found to decrease T cell proliferation in response to alloantigens. In a mouse model of skin transplantation, treatment with DPLG3 decreased the production of inflammatory cytokines and increased the expression of coinhibitory molecules by T cells. Next, DPLG3 was administered to mice that received a heart transplant from a mouse of another strain. These mice survived longer than the recipients that did not receive the compound, and there were few signs of rejection histologically. In addition, there were fewer effector T cells in the spleen, indicating the inhibition of an immune response. Overall, there was less lymphocyte infiltration and a decrease in T cell activation and proliferation compared to mice not treated with DPLG3. This study demonstrated that the synthesis of a selective proteasome inhibitor to prevent immune response to allografts could potentially allow for a safer and more efficacious medication for transplant recipients.

Click to read the study in PNAS

Relevant Reading: Proteasome inhibition suppresses essential immune functions of human CD4+ T cells

In-Depth [animal study]: An N,C-capped dipeptide, DPLG3, was synthesized to selectively bind to 5i 99,000-fold higher than 5c, a selectivity significantly greater than the currently available proteasome inhibitor. In vitro, DPLG3 inhibited 5i activity in Karpas lymphoma cells, while it had no effect on 5c activity in HepG2 human hepatoma cells. DPLG3 was not cytotoxic to human peripheral blood mononuclear cells. When T cells were isolated from the spleen of mice and treated with increasing concentrations of DPLG3, there was a concentration-dependent decrease in proliferation assessed using 3H-thymidine incorporation (p<0.05). A similar result was seen when mice that received a skin allograft were treated with 25 mg/kg of the compound. Their immune cells showed reduced proliferation in response to alloantigen, increased gene expression of coinhibitory molecules such as CTLA4 and BTLA, as well as a decrease in the pro-inflammatory cytokines IL-2 and IL-17 (p<0.05).

The efficacy of DPLG3 was then tested on C57BL/6 mice that were allografted with hearts from BALB/c mice. The recipient mice were then given 25 mg/kg DPLG3 each day for 14 days after receiving the transplant. These mice were found to have a significant decrease in the percentage of T cells in the spleen (p<0.05), demonstrating a lack of immune activation. In addition, the splenocytes demonstrated reduced responsiveness to alloimmune stimuli and there was a significant decrease in histologic signs of graft rejection. The treated mice had overall longer graft survival, with no death seen with 2 weeks of treatment with DPLG3 and a single dose of 25 g CTLA4 Ig.

Image: PD

20172 Minute Medicine, Inc. All rights reserved. No works may be reproduced without expressed written consent from 2 Minute Medicine, Inc. Inquire about licensing here. No article should be construed as medical advice and is not intended as such by the authors or by 2 Minute Medicine, Inc.

2 Minute Medicines The Classics in Medicine: Summaries of the Landmark Trials is available now in paperback and e-book editions.

This text summarizes the key trials in:General Medicine and Chronic Disease, Cardiology, Critical and Emergent Care, Endocrinology, Gastroenterology, Hematology and Oncology, Imaging, Infectious Disease, Nephrology, Neurology, Pediatrics, Psychiatry, Pulmonology, and Surgery.

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Air Pollution And Diabetes Study – 5newsonline.com

Posted: February 11, 2017 at 5:43 pm


5newsonline.com
Air Pollution And Diabetes Study
5newsonline.com
A new health study found that air pollution is linked to type two diabetes in overweight Latino children. The USC study looked at Latino children living in areas that were known to have high levels of nitrogen dioxide and other particulates in the air.

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How some insulin-producing cells survive in type 1 diabetes … – Science Daily

Posted: February 11, 2017 at 5:43 pm

How some insulin-producing cells survive in type 1 diabetes ...
Science Daily
Medical researches have identified how insulin-producing cells that are typically destroyed in type 1 diabetes can change in order to survive immune attack.
Yale study: Hope seen for producing insulin in type 1 diabeticsNew Haven Register

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Starting Insulin Earlier Can Limit Weight Gain for Type 2 Diabetes – Diabetes In Control

Posted: February 11, 2017 at 5:43 pm

Early initiation of therapy hoped to reduce long-term morbidity, mortality by getting to goal earlier; potentially alter the natural history of the disease.

We have seen that basal insulin causes less weight gain than other insulin regimes. In this prospective, multicenter analysis, we see data that suggests initiation of basal insulin therapy earlier on in disease duration may be beneficial for therapy on the grounds of concern over weight gain appears to be counter-productive. This is especially true given the potentially superior glycemic control also associated with early insulin initiation, further limiting weight gain. As such, prolonging the start of insulin.

Peter Bramlage, MD, of the Institute for Pharmacology and Preventive Medicine in Mahlow, Germany, and colleagues analyzed data from two groups of patients with type 2 diabetes diagnosed on or after Jan. 1, 2011, identified through the Diabetes Versorgungs-Evaluation (DIVE) registry, a German multicenter registry involving 200 physician offices specializing in type 2 diabetes. The first group included insulin-naive patients receiving basal insulin for the first time (n = 113; concomitant oral antidiabetic use was permitted); the second group included patients receiving their first oral antidiabetic therapy without simultaneous basal or short-acting insulin (n = 408).

Researchers found that, relative to baseline body weight, patients in the basal insulin group gained an average of 0.98 kg at 1 year vs. a loss of 1.52 kg for those not using insulin (P < .001); results persisted when expressed as a proportional change from baseline (P < .001).

In multivariable analysis, researchers observed that baseline weight (regression coefficient = 0.89; 95% CI, 0.81-0.97) and diabetes duration (regression coefficient = 2.52; 95% CI, 0.53-4.52) were the only factors that were predictors of weight gain between baseline and 1 year in the basal insulin group.

The researchers noted that the duration of diabetes before basal insulin therapy as an independent predictor of weight gain was logical, as early initiation would minimize HbA1c escalation and avoid the creation of a BMI deficit.

The researchers wrote that, Despite disagreement over the direction of weight change, findings from prior and present studies suggest that shorter diabetes duration is associated with more favorable weight outcomes, and early initiation of basal insulin therapy may be advantageous

Many of these clinical case studies exemplify the diversity of patients who may benefit from early insulin initiation. Ultimately, it is hoped that early initiation of therapy will not only prevent weight gain and short-term complications, but also reduce long-term morbidity and mortality by getting to goal earlier and potentially alter the natural history of the disease. This latter concept is currently of intense interest. Although optimal disease management is patient-specific, achieving and maintaining tight glycemic control are the primary goals of therapy. Because many type 2 diabetes patients will eventually require insulin therapy, overcoming fears and therapeutic barriers to initiating therapy early as needed are essential for reducing the vascular comorbidities of this highly prevalent disease in patients of all ages. Fortunately, a number of new clinical tools are available, including both prandial and basal insulin analogs, new insulin-delivery devices, and an ever-improving knowledge of the pathophysiology and natural history of diabetes.

Practice Pearls:

PLOS Feb.2nd, 2017

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Once Again, Periodontitis Tied to Diabetes Complications – Diabetes In Control

Posted: February 11, 2017 at 5:43 pm

Adults with type 2 diabetes and microvascular complications have an increased risk for severe periodontitis compared with those without microvascular complications.

Poor glycemic control is also associated with severity and prevalence of periodontitis in adults with type 2 diabetes, according to the researchers. The larger question is what comes first? Does diabetes cause periodontitis or does periodontitis increase our risk of diabetes? Over the years, Diabetes In Control has published at least 12-15 articles on the subject. (Do a search at Diabetesincontrol.com)

Shuji Inoue, MD, PhD, in Japan, and colleagues evaluated 620 adults with type 2 diabetes who presented to diabetes clinics at 21 institutions to determine associations between periodontitis, microvascular complications and glycemic control.

Overall, 34.5% of participants had retinopathy, 25.8% had nephropathy and 29.8% had neuropathy. Half of all participants had at least microvascular complication. Glycemic control was poor in 47.2% of participants, fair in 24.5%, good in 21.8% and excellent in 6.5%.

Compared with participants without microvascular complications, participants with all three microvascular complications had a higher prevalence of periodontitis (P < .01), but no significant difference was found for the prevalence of periodontitis between participants without microvascular complications and those with one or two.

Severe periodontitis was more prevalent in participants with three microvascular complications (49.4%) compared with those with one (37.8%) or two (36.9%). Participants with two or three microvascular complications had greater periodontitis severity compared with participants without complications.

Participants with poor glycemic control had a greater prevalence of periodontitis (71%) compared with participants with excellent glycemic control (50%; P < .01). Severe periodontitis was more prevalent in participants with poor glycemic control (40.6%) compared with those with fair glycemic control (28.3%), good glycemic control (28.1%) and excellent glycemic control (28%). Participants with good (P < .05), fair (P < .01) and poor glycemic control (P < .01) had greater periodontitis severity compared with participants with excellent glycemic control.

Research suggests that control of periodontal disease can play a key role in the control of diabetes. Thus, dentists must be aware of the signs and symptoms of diabetes, and understand the importance of maintaining periodontal health for anyone with diabetes.

Numerous studies have found a positive relationship between poor glycemic control in persons with type 2 and increased periodontitis. One five-year longitudinal study found increased attachment loss in adolescents with diabetes, whereas non-diabetic subjects had stable attachment levels.

A cross-sectional study of over 1,400 subjects found people with diabetes to have 2.3 times increased risk for attachment loss. In 48 studies on children and adolescents with type 1 diabetes, all but one found an increased prevalence of periodontal disease compared to children without diabetes. Of the eight reports limited to people with type 2, all found significantly poorer periodontal health in people with diabetes. In fact, after controlling for other risk factors, the odds of having periodontitis in people with diabetes was increased by two and a half to four times. Similar findings have been reported elsewhere.

In all cases, the level of diabetic control was a significant factor. Subjects with diabetes who were able to maintain consistent glycemic levels had no greater risk than did healthy subjects. For both type 1 and type 2 diabetes, there does not appear to be any correlation between the prevalence or severity of periodontal disease and the duration of diabetes.

The literature provides consistent evidence of greater prevalence and severity of periodontal disease in people with diabetes, both types 1 and 2. As these studies were conducted in distinctly different settings, with heterogeneous subjects and using a number of different measures of periodontal disease, we can state with confidence that people with diabetes have an increased susceptibility to periodontitis related to diabetes control.

Research suggests that control of periodontal disease plays a key role in the control of diabetes. Thus, dentists must be aware of the signs and symptoms of diabetes, and understand the importance of maintaining periodontal health for anyone with diabetes.

Practice Pearls:

Nitta H, et al. J Diabetes Investig. 2017;doi:10.1111/jdi.12633.

Firatli E. The relationship between clinical periodontal status and insulin-dependent diabetes mellitus. Results after 5 years. Journal of Periodontology 1997;68(2):136-40

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Biotechnology | University of Wisconsin River Falls

Posted: February 10, 2017 at 11:44 am

Mission Statement

The mission of the Biotechnology Program at the University of Wisconsin-River Falls is to provide its students with an education that establishes a strong foundation and appreciation for understanding developments in the rapidly advancing field of biotechnology, to develop the technical and critical thinking skills necessary for success in the field, to foster ethical behavior, and to promote outreach.

The field of modern biotechnology was born of molecular biology and biochemistry. Modern Biotechnology provides a set of tools that allow scientists to modify and harness the genetic capabilities of organisms. This has led to rapid advances in many areas including pharmaceutical development, agriculture, food microbiology, medical devices and environmental sciences.

Some examples of the products of biotechnology include herbicide, drought and insect resistant crops, drugs targeted specifically to disease processes resulting in fewer side effects, and bioremediation capable of removing greater amounts of environmental toxins at reduced cost.

The Biotechnology major at UWRF is an interdepartmental program with an emphasis on the molecular basis of life and the techniques utilized to study and control these processes under in vivo, in vitro, and commercial production conditions. UWRF LogoThe Biotechnology curriculum is an integrated sequence of courses selected from the curricula of the departments of Biology, Chemistry, Physics, Animal and Food Science, and Plant and Earth Science. It includes both traditional offerings of the departments involved and courses that reflect advances in biochemistry, biophysics, and molecular biology. The Biotechnology major is designed to provide students interested in pursuing careers in this rapidly expanding field with the academic background required to either secure entry level positions in industry or to continue their education in graduate or professional schools. A student may complete a B.S. degree in Biotechnology in the College of Arts and Sciences or the College of Agriculture, Food and Environmental Sciences.

Current curriculum check list (2008-2009)

Planning sheets

A scholarship has been established that is awarded to an outstanding junior or senior biotechnology major that either has worked on a research project, or will be participating in a research project during the year of the scholarship award. Follow the link above for information regarding scholarship criteria, recipients of the scholarship, and contributing to the scholarship fund.

Assessment of student learning is important to the University, the Colleges and the Biotechnology Program. Through appropriate assessment practices, we maintain a strong, current degree program and improve the quality of the education our students receive.

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Revolutionizing Biotechnology with Artificial Restriction Enzymes … – Genetic Engineering & Biotechnology News (press release)

Posted: February 10, 2017 at 11:44 am

Scientists at the University of Illinois say they have developed a new technique of genetic engineering for basic and applied biological research and medicine. Their work ("Programmable DNA-Guided Artificial Restriction Enzymes"), reported inACS Synthetic Biology,could open new doors in genomic research by improving the precision and adherence of sliced DNA, according to the investigators.

"Using our technology, we can create highly active artificial restriction enzymes with virtually any sequence specificity and defined sticky ends of varying length," said Huimin Zhao, Ph.,D., professor of chemical and biomolecular engineering, who leads a synthetic biology research group at the Carl R. Woese Institute for Genomic Biology at Illinois. "This is a rare example in biotechnology where a desired biological function or reagent can be readily and precisely designed in a rational manner."

Restriction enzymes cut DNA at a specific site and create a space wherein foreign DNA can be introduced for gene-editing purposes. This process is not achieved only by naturally occurring restriction enzymes; artificial restriction enzymes, or AREs, have risen to prominence in recent years. CRISPR/Cas9, a bacterial immune system used for "cut-and-paste" gene editing, and TALENs, or transcription activator-like effector nucleases, which are modified restriction enzymes, are two popular examples of such techniques.

Though useful in genetic engineering, no AREs generate defined "sticky ends"an uneven break in the DNA ladder structure that leaves complementary overhangs, improving adhesion when introducing new DNA. "If you can cleave two different DNA samples with the same restriction enzyme, the sticky ends that are generated are complementary," explained graduate student Behnam Enghiad. "They will hybridize with each other, and if you use a ligase, you can stick them together."

However, restriction enzymes themselves have a critical drawback: the recognition sequence that prompts them to cut is very short, usually only four to eight base pairs. Because the enzymes will cut anywhere that sequence appears, researchers rely on finding a restriction enzyme whose cut site appears only once in the genome of their organism or plasmid, an often difficult proposition when the DNA at hand might be thousands of base pairs long.

This problem has been partially solved simply by the sheer number of restriction enzymes discovered: more than 3600 have been characterized, and over 250 are commercially available. "Just in our freezer, for our other research, we have probably over 100 different restriction enzymes," said Enghiad. "We look through them all whenever we want to assemble something. The chance of finding the unique restriction site is so low."

"Our new technology unifies all of those restriction enzymes into a single system consisting of one protein and two DNA guides. Not only have you replaced them, but you can now target sites that no available restriction enzymes can."

The new method creates AREs through the use of an Argonaute protein (PfAgo) taken fromPyrococcus furiosus, an archeal species. Led by a DNA guide, PfAgo is able to recognize much longer sequences when finding its cut site, increasing specificity and removing much of the obstacles posed by restriction enzymes. Furthermore, PfAgo can create longer sticky ends than even restriction enzymes, a substantial benefit as compared to other AREs.

"When we started, I was inspired by a paper about a related proteinTtAgo. It could use a DNA guide to cleave DNA, but only up to 70 degrees," continued Enghiad. "DNA strands start to separate over 75 degrees, which could allow a protein to create sticky ends. If there were a protein that was active at higher temperatures, I reasoned, that protein could be used as an artificial restriction enzyme. SoI started looking for that, and what I found was PfAgo."

In addition to replacing restriction enzymes in genetic engineering processes, Enghiad and Dr. Zhao believe their technology will have broad applications in the biological research. By creating arbitrary sticky ends, PfAgo could make assembly of large DNA molecules easier and would enable cloning of large DNA molecules, such as biochemical pathways and large genes.

The application of these techniques is broad-reaching, they say, ranging from discovery of new small-molecule drugs to engineering of microbial cell factories for synthesis of fuels and chemicals to molecular diagnostics of genetic diseases and pathogens, which are the areas Dr. Zhao and Enghiad are currently exploring.

"Due to its unprecedented simplicity and programmability (a single protein plus DNA guides for targeting), as well as accessibility...we expect PfAgo-based AREs will become a powerful and indispensable tool in all restriction enzyme or nuclease-enabled biotechnological applications and fundamental biological research," predicts Dr. Zhao. "It is to molecular biology as the CRISPR technology is to cell biology."

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