New Publication in ACT – Time to Change the Clinical Trial Monitoring Paradigm: Results from a Multicenter Clinical Trial Using a Quality by Design Methodology, Risk-Based Monitoring and Real-Time Direct Data Entry


Our most recent paper entitled, Time to Change the Clinical Trial Monitoring Paradigm: Results from a Multicenter Clinical Trial Using a Quality by Design Methodology, Risk-Based Monitoring and Real-Time Direct Data Entry has been published in Applied Clinical Trials. It is coauthored with an independent QA expert, Michael Hamrell and one of our clients. We have other publications in ACT which are linked for your reference.


In the Innovator’s Prescription by Christensen, Grossman and Hwang, the authors share a concept that in order to maximize efficiencies, it is a good idea to bring the solution close to the problem that needs to be solved. Our approach to direct data entry at the time of the office visit and using computing and statistical tools to assess protocol compliance is surely consistent with that hypothesis. Writing data down first on a piece of paper creates an unnecessary gap of both space and time between the patient encounter and the study database.


We now have ongoing or completed 12 studies using direct data entry coupled with risk-based monitoring under 9 INDs/IDEs, and more are planned for this year. In addition to the US, study sites have included Canada and SE Asia. We expect at least one regulatory marketing application in 2014 and one in 2015. We have met with FDA and Health Canada and have made a presentation of our approach to EMA.


ON TARGET is the newsletter of Target Health Inc., a NYC-based contract research organization (CRO), providing strategic planning, regulatory affairs, clinical research, data management, biostatistics, medical writing and software services, including the paperless clinical trial, to the pharmaceutical and device industries.


For more information about Target Health contact Warren Pearlson (212-681-2100 ext. 104). For additional information about software tools for paperless clinical trials, please also feel free to contact Dr. Jules T. Mitchel or Ms. Joyce Hays. The Target Health software tools are designed to partner with both CROs and Sponsors. Please visit the Target Health Website.


Joyce Hays, Founder and Chief Editor of On Target

Jules Mitchel, Editor

Vanessa Hays, Editorial Contributor

Molecular Biology



Molecular biology is the branch of biology that deals with the molecular basis of biological activity. Molecular biology overlaps with other areas of biology and chemistry, particularly genetics and biochemistry. Molecular biology chiefly concerns itself with understanding the interactions between the various systems of a 1) ___, including the interactions between the different types of DNA, RNA and protein biosynthesis as well as learning how these interactions are regulated.


Writing in Nature in 1961, William Astbury described molecular biology as: “?not so much a technique as an approach, an approach from the viewpoint of the so-called basic sciences with the leading idea of searching below the large-scale manifestations of classical biology for the corresponding molecular plan. It is concerned particularly with the forms of biological molecules and is predominantly three-dimensional and structural – which does not mean, however, that it is merely a refinement of morphology. It must at the same time inquire into genesis and function.



Schematic relationship between biochemistry, genetics and molecular biology.


Researchers in molecular biology use specific techniques native to molecular biology but increasingly combine these with techniques and ideas from genetics and 2) ___. There is not a defined line between these disciplines. The figure above is a schematic that depicts one possible view of the relationship between the fields:


1. Biochemistry is the study of the chemical substances and vital processes occurring in living organisms. Biochemists focus heavily on the role, function, and structure of biomolecules. The study of the chemistry behind biological processes and the synthesis of biologically active molecules are examples of biochemistry.


2. Genetics is the study of the effect of genetic differences on organisms. This can often be inferred by the absence of a normal component (e.g. one gene). The study of “mutants“ – organisms which lack one or more functional components with respect to the so-called “wild type“ or normal phenotype. 3) ___ interactions (epistasis) can often confound simple interpretations of such “knockout“ studies.


3. Molecular biology is the study of molecular underpinnings of the processes of replication, transcription, translation, and cell function. The central dogma of molecular biology where genetic material is transcribed into RNA and then translated into protein, despite being an oversimplified picture of molecular 4) ___ biology, still provides a good starting point for understanding the field. This picture, however, is undergoing revision in light of emerging novel roles for RNA.


Much of the work in molecular biology is quantitative, and recently much work has been done at the interface of molecular biology and computer science in bioinformatics and computational biology. As of the early 2000s, the study of gene structure and function, molecular genetics, has been among the most prominent sub-field of 5) ___ biology. Increasingly many other loops of biology focus on molecules, either directly studying their interactions in their own right such as in cell biology and developmental biology, or indirectly, where the techniques of molecular biology are used to infer historical attributes of populations or species, as in fields in evolutionary biology such as population genetics and phylogenetics. There is also a long tradition of studying biomolecules “from the ground up“ in biophysics.


Since the late 1950s and early 1960s, molecular biologists have learned to characterize, isolate, and manipulate the molecular components of cells and 6) ___. These components include DNA, the repository of genetic information; RNA, a close relative of DNA whose functions range from serving as a temporary working copy of DNA to actual structural and enzymatic functions as well as a functional and structural part of the translational apparatus; and proteins, the major structural and enzymatic type of molecule in cells.


One of the most basic techniques of molecular biology to study protein function is expression cloning. In this technique, DNA coding for a protein of interest is cloned (using PCR and/or restriction enzymes) into a plasmid (known as an expression vector). A vector has 3 distinctive features: an origin of replication, a multiple cloning site (MCS), and a selective marker (usually antibiotic resistance). The origin of replication will have promoter regions upstream from the replication/transcription start site. This plasmid can be inserted into either bacterial or 7) ___ cells. Introducing DNA into bacterial cells can be done by transformation (via uptake of naked DNA), conjugation (via cell-cell contact) or by transduction (via viral vector). Introducing DNA into eukaryotic cells, such as animal cells, by physical or chemical means is called transfection.


Several different transfection techniques are available, such as calcium phosphate transfection, electroporation, microinjection and liposome transfection. DNA can also be introduced into eukaryotic cells using viruses or bacteria as carriers, the latter is sometimes called bactofection and in particular uses Agrobacterium tumefaciens. The plasmid may be integrated into the genome, resulting in a stable transfection, or may remain independent of the genome, called transient transfection. In either case, DNA coding for a protein of interest is now inside a cell, and the protein can now be expressed. A variety of systems, such as inducible promoters and specific cell-signaling factors, are available to help express the protein of interest at high levels. Large quantities of a protein can then be extracted from the bacterial or eukaryotic cell. The 8) ___ can be tested for enzymatic activity under a variety of situations, the protein may be crystallized so its tertiary structure can be studied, or, in the pharmaceutical industry, the activity of new drugs against the protein can be studied.


Polymerase chain reaction (PCR)

The polymerase chain reaction is an extremely versatile technique for copying DNA. In brief, PCR allows a single DNA sequence to be copied (millions of times), or altered in predetermined ways.


Gel electrophoresis

Gel electrophoresis is one of the principal tools of molecular biology. The basic principle is that DNA, RNA, and proteins can all be separated by means of an electric field and size. In agarose 9) ___ electrophoresis, DNA and RNA can be separated on the basis of size by running the DNA through an agarose gel. Proteins can be separated on the basis of size by using an SDS-PAGE gel, or on the basis of size and their electric charge by using what is known as a 2D gel electrophoresis.


Macromolecule blotting and probing

The terms northern, western and eastern blotting are derived from what initially was a molecular biology joke that played on the term Southern blotting, after the technique described by Edwin Southern for the hybridisation of blotted DNA. Patricia Thomas, developer of the RNA blot which then became known as the northern blot, actually didn’t use the term. Further combinations of these techniques produced such terms as southwesterns (protein-DNA hybridizations), northwesterns (to detect protein-RNA interactions) and farwesterns (protein-protein interactions), all of which are presently found in the literature.


Southern blotting

Named after its inventor, biologist Edwin Southern, the Southern blot is a method for probing for the presence of a specific DNA sequence within a DNA sample. DNA samples before or after restriction enzyme (restriction endonuclease) digestion are separated by gel electrophoresis and then transferred to a membrane by blotting via capillary action. The membrane is then exposed to a labeled DNA probe that has a complement base sequence to the sequence on the DNA of interest. Most original protocols used radioactive labels, however non-radioactive alternatives are now available.


These blots are still used for some applications, however, such as measuring transgene copy number in transgenic mice, or in the engineering of gene knockout embryonic stem cell lines. A DNA array is a collection of spots attached to a solid support such as a microscope slide where each spot contains one or more single-stranded DNA oligonucleotide fragment. Arrays make it possible to put down large quantities of very small (100 micrometre diameter) spots on a single slide. Each spot has a DNA fragment molecule that is complementary to a single DNA sequence (similar to Southern blotting). Arrays can also be made with molecules other than DNA. For example, an antibody array can be used to determine what proteins or bacteria are present in a blood sample.


In molecular biology, procedures and technologies are continually being developed and older technologies abandoned. For example, before the advent of DNA gel electrophoresis (agarose or polyacrylamide), the size of DNA molecules was typically determined by rate sedimentation in sucrose gradients, a slow and labor-intensive technique requiring expensive instrumentation; prior to sucrose gradients, viscometry was used.


Aside from their historical interest, it is often worth knowing about older technology, as it is occasionally useful to solve another new problem for which the newer technique is inappropriate.


History of molecular biology

While molecular biology was established in the 1930s, the term was coined by Warren Weaver in 1938. Weaver was the director of Natural Sciences for the Rockefeller Foundation at the time and believed that biology was about to undergo a period of significant change given recent advances in fields such as X-ray crystallography. He therefore channeled significant amounts of (Rockefeller Institute) money into biological fields.


Clinical research and medical therapies arising from molecular biology are partly covered under gene therapy. The use of molecular biology or molecular cell biology approaches in medicine is now called molecular medicine. Molecular biology also plays important role in understanding formations, actions, regulations of various parts of cells which can be used efficiently for targeting new 10) ___, diagnosis of disease, physiology of the Cell.


ANSWERS: 1) cell, 2) biochemistry; 3) Genetic; 4) biology; 5) molecular; 6) organisms; 7) animal; 8) protein; 9) gel; 10) drugs

Eric Lander (1957 to Present)


Eric Lander has a Ph.D. in pure mathematics, in a subfield so esoteric and specialized that even if someone gets a great result, it can be appreciated by only a few dozen people in the entire world. But he left that world behind and, with no formal training, entered another: the world of molecular biology, medicine and genomics.



FIRST PLACE Eric Lander, victorious at the 1974 Science Talent Search. The same year he made the American team in the Mathematics Olympiad.


Eric Steven Lander (born February 3, 1957) is a Professor of Biology at the Massachusetts Institute of Technology (MIT), former member of the Whitehead Institute, and founding director of the Broad Institute of MIT and Harvard who has devoted his career to realizing the promise of the human genome for medicine. He is co-chair of U.S. President Barack Obama’s Council of Advisors on Science and Technology. In 2013 he was awarded the $3 million Breakthrough Prize in Life Sciences for his work. Lander also teaches freshman biology (a course he never took) at M.I.T., and runs a lab.


Eric Lander – as a friend, Prof. David Botstein of Princeton, put it – knows how to spot and seize an opportunity when one arises. And he has another quality, says his high school friend Paul Zeitz: bravery combined with optimism. “He was super smart, but so what?“ said Dr. Zeitz, now a mathematics professor at the University of San Francisco. “Pure intellectual heft is like someone who can bench-press a thousand pounds. But so what, if you don’t know what to do with it?“ Eric Lander, he added, knew what to do. And he knew how to carry out strong ideas about where progress in medicine will come from – large interdisciplinary teams collaborating rather than single researchers burrowed in their labs.


A Math Club Standout

Eric Steven Lander grew up in Flatlands, a working-class neighborhood in Brooklyn, raised by his mother – his father died of multiple sclerosis when Eric was 11. “Nobody in the neighborhood was a scientist,“ Dr. Lander said. “Very few had gone to college.“ His life changed when he took an entrance exam and was accepted at the elite Stuyvesant High School in Manhattan. He joined the math team and loved it – the esprit de corps, the competition with other schools, the social aspect of being on the team. “I found other kids, ninth graders, who also loved math and loved having fun,“ he said. He was so good that he was chosen for the American team in the 1974 Mathematics Olympiad. To prepare, the team spent a summer training at Rutgers University in New Brunswick, N.J. This was the first time the United States had entered the competition, and the coaches were afraid the team would be decimated by entrants from Communist countries. (Indeed, the Soviet Union placed first, but the Americans came in second, just ahead of Hungary, which was known for its mathematics talent.)


Dr. Zeitz was Dr. Lander’s roommate that summer. The two recall being the only teammates who did not come from affluent suburban families, and who did not have fathers. But Eric stood out for other reasons. “He was outgoing,“ Dr. Zeitz recalled. “He was, compared to the rest of us, definitely more ambitious. He was enthusiastic about everything. And he had a real charisma.“ Team members decided that Dr. Lander was the only one among them whom they could imagine becoming a United States senator one day. At first, though, it looked as if the young mathematician would follow a traditional academic path. He went to Princeton, majoring in mathematics but also indulging a passion for writing. He took a course in narrative nonfiction with the author John McPhee and wrote for the campus newspaper.


He graduated as valedictorian at age 20, won a Rhodes scholarship, went to Oxford and earned a mathematics Ph.D. there in record time – two years. Yet he was unsettled by the idea of spending the rest of his life as a mathematician. “I began to appreciate that the career of mathematics is rather monastic,“ Dr. Lander said. “Even though mathematics was beautiful and I loved it, I wasn’t a very good monk.“ He craved a more social environment, more interactions. “I found an old professor of mine and said, ?What can I do that makes some use of my talents?’“ He ended up at Harvard Business School, teaching managerial economics. He had never studied the subject, he confesses, but taught himself as he went along. “I learned it faster than the students did,“ Dr. Lander said.


At 23, he spoke to his brother, Arthur, a neurobiologist, who sent him mathematical models of how the cerebellum worked. The models “seemed hokey,“ Dr. Lander said, “but the brain was interesting.“ His appetite for biology whetted, he began hanging around a fruit-fly genetics lab at Harvard. A few years later, he talked the business school into giving him a leave of absence. He told Harvard he would go to M.I.T., probably to learn about artificial intelligence. Instead, he ended up spending his time in Robert Horvitz’s worm genetics lab. And that led to the spark that changed his life.


Making the Leap

It was 1983, and while Dr. Lander was hanging around the worm lab, Dr. Botstein, at the time a professor at M.I.T., was growing increasingly frustrated. He had spent five fruitless years looking for someone who knew mathematics to take on a project involving traits like high blood pressure that were associated with multiple genes. For these diseases, the old techniques for finding traits caused by single genes would not work. “I literally went around looking for someone who could help,“ Dr. Botstein said. Finally, at a conference, another biologist said, “There’s this fellow, Lander, at Harvard Business School who wanted to do something with biology.“ Dr. Botstein hunted Dr. Lander down at a seminar at M.I.T., and pounced. The two connected immediately. “We went to a whiteboard,“ Dr. Lander said, “and started arguing.“ Within a week, Dr. Lander had solved the problem. Then the two researchers invented a computer algorithm to analyze maps of genes in minutes instead of months. Soon, Dr. Lander had immersed himself in problems of mapping human disease genes. He had long discussions with Dr. Botstein about the future of human genomics. It was a time, Dr. Botstein said, “when talk of sequencing the human genome was just beginning to get traction.“ Dr. Lander wanted to know if there was any use for a mathematician in biology, and Dr. Botstein, who knew the challenges ahead, assured him there was. “He had a sufficiently high opinion of himself,“ Dr. Botstein said. “He thought that if anyone could do it, he could. He took a chance and dropped his Harvard job. It was clear that teaching economics would no longer be his career path.“


David Baltimore, a Nobel laureate who was then the head of the Whitehead Institute for Biomedical Research at M.I.T., was taken with Dr. Lander’s passion and abilities. He enabled Dr. Lander to become a fellow there and then an assistant professor in 1986. That same year, Dr. Lander went to a meeting at the Cold Spring Harbor Laboratory on Long Island where leading scientists held the first public debate on the idea of mapping the human genome. Dr. Lander raised his hand and joined the discussion, impressing the others so much they invited him into their circle. “It is very easy to be an expert in a new field where there are no experts,“ Dr. Lander said. “All you have to do is raise your hand.“ Meanwhile, Dr. Botstein and Dr. Baltimore wrote to the MacArthur Foundation recommending Dr. Lander for a “genius“ grant. He received it in 1987. He was 30. “I tried to help him over the years in realizing his dreams.“ Dr. Baltimore said. “And he’s been very successful in making that happen.“ Soon, Dr. Lander had become a central figure in the effort to sequence the human genome, leading the largest of the three centers that did most of the work. He combined his mathematics and the biology and chemistry he’d learned hanging out in labs. And he added insights about industrial organization, achieved in his business school days, to streamline the effort and control costs. What he loved most about the work was the community he had craved, the team effort he had been searching for.


Even before the Human Genome Project ended, Dr. Lander was thinking of how to keep what he saw as a wonderful collaboration among scientists going. There were, by his count, about 65 collaborations among young scientists in Cambridge and Boston, all outside the usual channels.

“Something magical had happened,“ Dr. Lander said. “People were coming together and taking on really bold problems.“ It may have had something to do with Dr. Lander’s personality. Gus Cervini, an administrator at Brigham and Women’s Hospital in Boston who worked for him for four years, used to call him “the sun.“ “He has this amazing influence or power on people,“ Mr. Cervini said. “He had this ability to get people to really think big. “When the sun shines on you, you feel like you can do anything.“


Persistence Rewarded

That power may have helped when Dr. Lander approached the presidents of Harvard and M.I.T. and proposed creating a permanent institute to continue the collaborative process that groups of scientists had been improvising. At first, he met with resistance, but he persisted. Then Dr. Baltimore introduced him to the philanthropists Eli and Edythe Broad, who had made their fortune in real estate. The Broads (the name rhymes with code) visited Dr. Lander’s lab one Saturday morning in October 2002. A few months later, they agreed to invest $10 million a year for a decade, so Dr. Lander could start what he thought of as an experiment with a new kind of research institute. The Broad Institute was to become a joint effort between Harvard and M.I.T., headed by Dr. Lander, that would encourage scientists to collaborate to solve big problems in biology, genetics and genomics. Within 18 months, the Broads doubled their gift, to $200 million. In 2008, they contributed another $400 million as an endowment to make the institute permanent. Today the institute has about 1,800 collaborating scientists from the two universities and Harvard’s hospitals.


Its aims sound audacious: “Assemble a complete picture of the molecular components of life. Define the biological circuits that underlie cellular responses. Uncover the molecular basis of major inherited diseases. Unearth all the mutations that underlie different cancer types. Discover the molecular basis of major infectious diseases. Transform the process of therapeutic discovery and development.“ “Half the place is devoted to finding the basis of disease and half is devoted to trying to transform and accelerate the development of therapeutics,“ Dr. Lander said. “It’s different from what you find in many university settings where you have many labs, each of whom does its own thing.“ The Broad is an experiment, Dr. Lander said, one that involves an institution and how to do scientific research. “This is in a sense a protected space to see if it works,“ Dr. Lander said.


The institute is Dr. Lander’s passion, but hardly his only one. His days start and end in a gym on the second floor of his house, where he has an elliptical cross-trainer. He uses it for two 40-minute sessions, one in the morning and one at night, watching Netflix videos and burning – according to the machine – 1,000 calories a day. He reports that he lost 42 pounds last summer without changing his diet. They bought the place, a converted schoolhouse, when his wife, Lori Lander, who is an artist, pointed out that it had a basketball court on the top floor – it could be a kind of neighborhood hangout, so the Landers would always know where their three children were. After his morning workout, he sometimes goes to a local bakery where he can work quietly. He arrives at the Broad between 8 and 10 a.m. In the fall, he teaches introductory biology to a class of 700 M.I.T. students on Monday, Wednesday and Friday mornings. He often meets with graduate students and postdoctoral fellows in the afternoon to discuss their work. Then he has his administrative duties and his meetings with philanthropists, trying to raise more money. He also spends 20% of his time in yet another role, as co-chairman of President Obama’s Council of Advisers on Science and Technology, which deals with topics like influenza vaccines, health information technology, science education and energy policy. In the evening, around 6:30 or 7, he has dinner with his family. His wife cooks – Dr. Lander loves to cook but says he just does not have time. He also reads – fiction, nonfiction, New Yorker articles – but has no patience with poor writing. “I am very eclectic in my reading, but it has to be really well written,“ he said. “That’s a huge barrier.“ On weekends he and his wife try to get to New York for the theater, another of his passions.


And he marvels at how his life has turned out. “I feel like it’s so incredibly lucky to end up here,“ he said. “I could not have planned this. What if I hadn’t met David Botstein? What if I hadn’t gone to a meeting where the human genome was discussed? I have no idea. This is as random as it gets. “It’s a very weird career.“Source: The New York Times; Wikipedia

Cognitive Training Shows Staying Power


According to an article published in the Journal of the American Geriatrics Society (2014; 62:6-24), training to improve cognitive abilities in older people lasted to some degree 10 years after the training program was completed. The report, from the Advanced Cognitive Training for Independent and Vital Elderly (ACTIVE) study, showed training gains for aspects of cognition involved in the ability to think and learn. However, the authors said that memory training did not have an effect after 10 years.


The original 2,832 volunteers for the ACTIVE study were divided into three training groups — memory, reasoning and speed-of-processing — and a control group. The training groups participated in 10, 60- to 70-minute sessions over five to six weeks, with some randomly selected for later booster sessions. The study measured effects for each specific cognitive ability trained immediately following the sessions and at one, two, three, five and 10 years after the training.


The investigators were also interested in whether the training had an effect on the participants’ abilities to undertake some everyday and complex tasks of daily living. They assessed these using standardized measures of time and efficiency in performing daily activities, as well as asking the participants to report on their ability to carry out everyday tasks ranging from preparing meals, housework, finances, health care, using the telephone, shopping, travel and needing assistance in dressing, personal hygiene and bathing.


At the end of the trial, all groups showed declines from their baseline tests in memory, reasoning and speed of processing. However, the participants who had training in reasoning and speed of processing experienced less decline than those in the memory and control groups. Results of the cognitive tests after 10 years show that 73.6% of reasoning-trained participants were still performing reasoning tasks above their pre-trial baseline level compared to 61.7% of control participants, who received no training and were only benefiting from practice on the test. This same pattern was seen in speed training: 70.7% of speed-trained participants were performing at or above their baseline level compared to 48.8% of controls. There was no difference in memory performance between the memory group and the control group after 10 years. Participants in all training groups said they had less difficulty performing the everyday tasks compared with those in the control group. However, standard tests of function conducted by the researchers showed no difference in functional abilities among the groups.


The ACTIVE study followed healthy, community-dwelling older adults from six cities-Baltimore; Birmingham, Ala.; Boston; Detroit; State College, Pa.; and Indianapolis. The participants averaged 74 years of age at the beginning of the study and 14 years of education, 76% were female, 74% were white and 26% were African-American. The 10-year follow-up was conducted with 44% of the original sample between April 1998 and October 2010.

Enzyme that Produces Melatonin Originated 500 Million Years Ago


Melatonin is a key hormone that regulates the body’s day and night cycle. it is manufactured in the brain’s pineal gland and is found in small amounts in the retina of the eye. Melatonin is produced from the hormone serotonin, the end result of a multistep sequence of chemical reactions. The next-to-last step in the assembly process consists of attaching a small molecule — the acetyl group — to the nearly finished melatonin molecule. This step is performed by an enzyme called arylalkylamine N-acetyltransferase, or AANAT. Because of its key role in producing the body clock-regulating melatonin, AANAT is often referred to as the timezyme.




According to an article published online in PNAS (December 2013), an international team of scientists led by senior author David C. Klein, Ph.D., Chief of the Section on Neuroendocrinology in the NIH’s Eunice Kennedy Shriver National Institute of Child Health and Human Development (NICHD) and at institutions in France, Norway, and Japan, has traced the likely origin of the enzyme needed to manufacture the hormone melatonin to roughly 500 million years ago. The work indicates that this crucial enzyme, which plays an essential role in regulating the body’s internal clock, likely began its role in timekeeping when vertebrates (animals with spinal columns) diverged from their nonvertebrate ancestors. According to the authors, an understanding of the enzyme’s function before and after the divergence may contribute to an understanding of such melatonin-related conditions as seasonal affective disorder, jet lag, and to the understanding of disorders involving vision. The findings also provide strong support for the theory that the time-keeping enzyme originated to remove toxic compounds from the eye and then gradually morphed into the master switch for controlling the body’s 24-hour cyclic changes in function. The authors also isolated a second, nonvertebrate form of the enzyme from sharks and other contemporary animals thought to resemble the prototypical early vertebrates that lived 500 million years ago.


According to the authors, the form of AANAT found in vertebrates occurs in the brain’s pineal gland and, in small amounts, in the retina. Another form of the enzyme, termed nonvertebrate AANAT, has been found only in other forms of life, such as bacteria, plants and insects. Interestingly, nonvertebrate AANAT appears to detoxify a broad range of potentially toxic chemicals,. In contrast, vertebrate AANAT is highly specialized for adding an acetyl group to melatonin. According to Dr. Klein, “the two are as different from each another as a Ferrari is from a Model-T Ford, considering the speed of the reaction and how fast it can be turned on and off.“


In 2004, Dr. Klein and his coworkers published a theory that melatonin was at first a kind of cellular waste, a by-product created in cells of the eye when normally toxic substances were rendered harmless. Because melatonin accumulated at night, the ancestors of today’s vertebrates became dependent on melatonin as a signal of darkness. As the need for greater quantities of melatonin grew, the pineal gland developed as a structure separate from the eyes, to keep serotonin and other toxic substances needed to make melatonin away from sensitive eye tissue.


“The pineal glands of birds and reptiles can detect light,“ Dr. Klein said. “And the retinas of human beings and other species also make melatonin. So it would appear that both tissues evolved from a common, ancestral, light-detecting tissue.“ Before the current study, the researchers lacked proof of their theory, particularly in regard to the question of how the vertebrate form of the enzyme originated because it did not appear to exist in non-vertebrates and had been found only in bony fishes, reptiles, birds, and mammals – all of which lacked the non-vertebrate form. The first evidence of how the vertebrate form of the enzyme originated came when study co-author Steven L. Coon, also of NICHD, discovered genes for the nonvertebrate and vertebrate forms of AANAT in genomic sequences from the elephant shark, considered to be a living representative of early vertebrates. This finding indicated that the vertebrate form of AANAT may have resulted after a phenomenon known as gene duplication. Gene duplication typically results from any of a number of genetic mishaps during cell division. Instead of one copy of a gene resulting from the process, an additional copy results, so that there are two versions of a gene where only one existed previously. The phenomenon is thought to be a major factor influencing evolutionary change.


The authors theorized that following duplication, one form of AANAT remained unchanged and the other gradually evolved into the vertebrate form. The authors hypothesized that at some point after vertebrate AANAT developed, vertebrates appear to have stopped making the nonvertebrate form, perhaps because it was no longer needed or because its function was replaced by a similar enzyme.


Before the authors could continue, they needed to confirm their finding, to rule out that the nonvertebrate AANAT they found didn’t result from accidental contamination with bacteria or some other organism. As a result, DNA from Mediterranean sharks and sea lampreys was obtained via fishermen’s catches by Jack Falcon of the Arago Laboratory, a marine biology facility that is part of the CNRS and the Pierre and Marie Curie University in France. Samples from a close relative of the elephant shark — the ratfish — were provided by Even-Jorgensen at the Arctic University of Norway. Finally, Susumo Hyodo of the University of Tokyo contributed samples from elephant sharks he collected off the coast of Australia. Next, the Hyodo and Falcon groups isolated RNA from the retinas and pineal glands of the animals. RNA is used to direct the assembly of amino acids into proteins. From these RNA sequences, it was possible to assemble working versions of AANAT molecules — both the vertebrate and nonvertebrate forms.


The sequences of the proteins encoded by the AANAT genes were analyzed by Eugene Koonin and Yuri Wolf of the National Library of Medicine using computer techniques designed to study evolution. Peter Steinbach, of NIH’s Center for Information Technology, examined the three-dimensional structures of nonvertebrate and vertebrate AANAT in the study animals and determined that the two forms of the enzyme likely had a common ancestor.


Taken together, their results provide evidence for the hypothesis that nonvertebrate AANAT resulted from duplication of the non-vertebrate AANAT gene about 500 million years ago and that following this event one copy of the duplicated gene eventually changed into the gene for vertebrate AANAT. In addition to providing information on the origin of melatonin and the evolution of AANAT, the findings also have implications for research on disorders affecting vision. Vertebrate AANAT and melatonin are found in small amounts in the eyes of humans and other vertebrates. Although they may play a role in detoxifying compounds, it is also reasonable to consider that this detoxifying function is shared with other enzymes.

TARGET HEALTH excels in Regulatory Affairs. Each week we highlight new information in this challenging area.


FDA Allows Marketing for First of-its-Kind Post-Natal Test to Help Diagnose Developmental Delays and Intellectual Disabilities in Children


According to the National Institutes of Health and the American Academy of Pediatrics, 2-3% of children in the US have some form of intellectual disability. Many intellectual and developmental disabilities, such as Down syndrome and DiGeorge syndrome, are associated with chromosomal variations.


The FDA has authorized for marketing the Affymetrix CytoScan Dx Assay, which can detect chromosomal variations that may be responsible for a child’s developmental delay or intellectual disability. Based on a blood sample, the test can analyze the entire genome at one time and detect large and small chromosomal changes.


The FDA reviewed the Affymetrix CytoScan Dx Assay through its de novo classification process, a regulatory pathway for some novel low-moderate-risk medical devices. For the de novo petition, the FDA’s review of the CytoScan Dx Assay included an analytical evaluation of the test’s ability to accurately detect numerous chromosomal variations of different types, sizes, and genome locations when compared to several analytically validated test methods. The FDA found that the CytoScan Dx Assay could analyze a patient’s entire genome and adequately detect chromosome variations in regions of the genome associated with intellectual and developmental disabilities.


Additionally, the agency’s review included a study that compared the performance of the CytoScan Dx Assay to tests that are commonly used for detecting chromosomal variations associated with a developmental delay or intellectual disability. A comparison of test results from 960 blood specimens showed the CytoScan Dx had improved ability over commonly used tests, including karyotyping and FISH chromosomal tests, to detect certain chromosomal abnormalities.


This device should not be used for stand-alone diagnostic purposes, pre-implantation or prenatal testing or screening, population screening, or for the detection of, or screening for acquired or genetic aberrations occurring after birth, such as cancer. The test results should only be used in conjunction with other clinical and diagnostic findings, consistent with professional standards of practice, including confirmation by alternative methods, evaluation of parental samples, clinical genetic evaluation, and counseling as appropriate. Interpretation of test results is intended to be performed only by health care professionals who are board certified in clinical cytogenetics or molecular genetics.


Affymetrix CytoScan Dx Assay is manufactured by Affymetrix, Inc., located in Santa Clara, Calif.

Thai Sweet Potatoes with Halibut and Ginger-Lime Peanut Sauce


Sweet Potatoes & Halibut before adding the Peanut Sauce, Photo: ©Joyce Hays, Target Health Inc.



Sweet Potato on left, Halibut on right, with Peanut sauce drizzled over both. Photo: ©Joyce Hays, Target Health Inc.



Buy very fresh halibut. Figure 1 lb fillet (okay to buy in small pieces) for every two people. Prepare the halibut in your own way, just before serving. Place an individual portion of fish, centered on a plate, now put a filled half sweet potato, over part of the fish and spoon the sauce over fish and potato. Garnish with a small amount of shredded coconut & pinch of cilantro. We did the halibut in the photo, by dipping the cleaned halibut first, both sides of the fish, into a beaten egg mixture, then into Panko crumbs; then into 1 Tablespoon hot olive oil mixed with 1 Tablespoon chicken stock (or canola) in fry pan, getting the Panko a golden brown on each side, removing to serving dish, placed into a warming drawer.





  • 3 medium-sized sweet potatoes
  • 3 Tablespoons olive oil + extra to rub on potatoes
  • 1 heaping Tablespoon shredded coconut
  • 1 onion, chopped
  • 1 cup fresh sliced cremini mushrooms
  • 3 cloves garlic, minced
  • 1 teaspoon turmeric
  • 1 bunch kale, cleaned, stems removed, any hard veins cut out
  • 1/2 cup chicken stock or broth
  • 2/3 cup fresh cilantro, chopped
  • 1 cup salt-free peanuts, chopped



  • 3/4 cup hot chicken broth
  • 1 garlic clove, juiced
  • 3/4 cup chunky Peanut Butter
  • 1 teaspoon turmeric
  • 2 Tablespoons tahini
  • 2 Tablespoons freshly squeezed lime juice (1 to 2 limes)
  • 1 Tablespoon fresh grated ginger



1/2 cup shredded coconut; pinch chopped peanuts, pinch chopped cilantro



1. Preheat oven to 4000F.

2. Scrub potatoes well.

3. With a paper towel, rub each potato with olive oil.


Bake sweet potatoes 30 to 40 minutes, until a knife easily goes through the potatoes. Remove potatoes from oven and set aside. When potatoes are cooler, cut them in half and set aside. Leave the oven on.


Roll up the Kale leaves, then slice them thinly, to get thin julienned kale strips


To make the filling:

In a saute pan over medium heat, add the olive oil, onions and kale strips cook 3 to 4 minutes, until golden brown. Stir in mushrooms and garlic, cook 2 minutes longer. Now, stir in the cilantro, turmeric, coconut and chopped peanuts and cook another 2 minutes, while stirring constantly. Finally, scoop out the flesh of each potato, carefully, so you don’t break the skins and set the skins aside in an oiled baking dish. Stir all ingredients together.


Now, fill each skin half with the potato mixture and place back into the oiled baking dish.  Keep warm in a warming drawer or low-heat oven, until ready to serve.


To make the Peanut sauce:

Whisk together peanut butter, tahini, lime juice, garlic and ginger. Add the hot chicken broth last, very slowly as you whisk, to create a thick sauce and not a runny thin sauce. Set aside. If sauce needs to be thicker, add 1 teaspoon agar and mix well.


Serve sweet potato and fish drizzled with extra peanut sauce and topped with chopped peanuts pinch cilantro, pinch shredded coconut



We clinked glasses to our Anniversary this past Friday, and celebrated with these recipes, preceded by one of many kale salad recipes. We both agree that the addition of the peanut sauce, really makes the sweet potato and halibut sing out. It has taken about a year to perfect the Peanut sauce, because the peanut flavor is so overpowering. The final version, shared with you, retains a faint nutty flavor, that when combined with tahini, turmeric, ginger, lime and garlic, is totally different from a year ago, when I first started to experiment with a peanut sauce for fish, chicken, or veggies. (And you know who, was the amiable, but stern critic guinea pig for a year)


Cheers to staying married!


To Health, Love, Friendship, and Successful Achievement in 2014! Photo: ©Joyce Hays, Target Health Inc.