Ex-Officio Council Members Present:

Colonel Kenneth J. Hoffman, M.D., M.P.H.
Richard T. Suchinsky, M.D.

Chairperson:     Ting-Kai Li, M.D.

Executive Secretary:    Kenneth R. Warren, Ph.D.

Council Assistants:       Mrs. Ida Nestorio and Ms. M. Virginia Wills

Approximately 140 additional observers attended the open session, including representatives from constituency groups, liaison organizations, NIAAA staff, and members of the general public.

Dr. Ting-Kai Li, Director, NIAAA, called the closed session of the 103^rd meeting of the NACAAA to order at 5:30 p.m., June 4, 2003, for consideration of grant applications.

Dr. Kenneth R. Warren, Director, Office of Scientific Affairs, reviewed procedures for critiquing grant applications and reminded Council members of regulations pertaining to conflict of interest and confidentiality. Members absented themselves from the discussion and evaluation of applications from their own institutions and in situations involving any real, apparent, or potential conflict of interest.

Dr. Kenneth Warren, Council Executive Secretary, called the open session of the 103^rd meeting of the NACAAA to order at 8:50 a.m., explaining that Dr. Li was called to a concurrent meeting of other Institute Directors and would join the Council meeting at mid-morning. He also announced that three Council members were unable to attend: Dr. Steve Mirin, Dr. Richard Deitrich, and Sheryl Ramstad.

Dr. Warren introduced the newly appointed Council member, Mrs. Hope Taft, First Lady of the State of Ohio and a recognized leader for over 25 years in building coalitions of citizens and service organizations that focus on substance abuse prevention issues.

Former Council member, Dr. Mark Goldman, was also welcomed as a new Associate Director of NIAAA with responsibilities for integrating the Institute’s behavioral and biomedical research as well as developing a spin-off of the college drinking initiative that targets the full spectrum of underage drinking. Dr. Goldman was formerly Director of the Alcohol and Substance Abuse Research Institute at the University of South Florida and he also co-chaired the NIAAA Advisory Council’s Task Force on College Drinking.

Referring to the formal Director's Report in Council members’ folders, Dr. Warren noted its new format and abbreviated content and announced that Ms. Charlotte Armstrong has assumed responsibility for compiling this document. He also praised the many efforts that Mrs. Brenda Hewitt has made to this endeavor over many years.

The following Institute activities were highlighted:

1. The omnibus FY 2003 appropriation bill that was signed on February 21^st allotted $416,051,000 to NIAAA—an 8.6 percent increase over FY 2002. The monies will support 192 competing grants; 15 previously approved research centers; and 247 full-time training positions—with a 4 percent increase in trainee stipends. NIAAA’s FY 2002 budget, FY 2003 appropriations, and FY 2004 budget request are itemized in the Report.
   
   
2. An April 28^th meeting of the Leadership to Keep Children Alcohol Free was attended by such dignitaries as the DHHS Secretary, NIH Director, Administrator of the Substance Abuse and Mental Health Services Administration, as well as NIAAA leaders and Governors’ spouses. The U.S. Surgeon General, Dr. Richard Carmona, responded enthusiastically to requests to 1) include childhood drinking prevention in his presentations around the country, 2) invite Governors' spouses on his 50-State/50-school tour, 3) assist in the creation of a public service announcement, and 4) produce a special Surgeon General’s Report on childhood drinking.
   
   
3. National Alcohol Screening Day—on April 10^th was the most successful ever, with a total of 4,624 sites offering screening activities, brief interventions, and referrals that reflected the theme, Alcohol and Your Health: Where Do You Draw the Line? The event drew good media coverage, with articles in USA Today, Parade Magazine, and the columns of Dear Abby, Annie's Mailbox, and Hints from Heloise.
   
   
4. A newly revised, 22-page booklet, Helping Patients with Alcohol Problems: A Health Practitioner’s Guide, was re-issued and disseminated to thousands of sites nationwide on National Alcohol Screening Day. The American Medical Association also mailed the publication to 40,000 members with a cover letter signed by the Chair of its Board of Trustees and Dr. Li.
   
   
5. NIAAA joined six other NIH institutes on April 16^th to celebrate the 50^th anniversary of Drs. Watson and Crick's landmark paper on the structure of DNA. At a scientific symposium entitled, Genes, Brain, Behavior: Before and Beyond Genomics, Council member and NIAAA grantee, Dr. Howard Edenberg, discussed his research on the risk factors for alcoholism. NIAAA also co-sponsored another April 16^th symposium with the National Institute on Environmental Health Sciences and the National Human Genome Research Institute entitled, Genetic Variation and Gene/Environment Interactions in Human Health and Disease.
   
   

NIAAA Director, Dr. Li, appeared before a number of groups, including:

• On May 11^th, Dr. Li was presented with an honorary Doctorate of Science degree from Indiana University where he had served for over 30 years. In bestowing the degree, Dr. Gerald Bepko, Interim President, noted that Dr. Li exemplifies "the highest standards of professional excellence and an unswerving commitment to the aims of scientific research in the public good."
  
  
• As part of media coverage for Alcohol Screening Day, Dr. Judith Arroyo, NIAAA Office of Collaborative Research, gave an interview in Spanish that was broadcast on a Spanish-language show on April 2^nd. Additionally, an op-ed article by First Ladies Mary Easley and Hope Taft appeared in the Washington Times on April 27^th expressing their concern about recently published findings by the Centers for Disease Control that 25 percent of all 9^th graders,14- to15- years-old, acknowledge binge drinking (5 or more drinks in one session) within the past month.
  
  
• Recent NIAAA-Sponsored Workshops/Symposia focused on pharmacotherapy for smoking cessation, spirituality and health, a core assessment methodology for clinical trials, medications to treat alcohol use disorders, and—with the National Heart, Lung and Blood Institute, Alcohol and Cardiovascular Systems Research: Challenges and Opportunities.

• Among materials NIAAA produced for Alcohol Screening Day are two pamphlets entitled, A Family History of Alcoholism: Are You At Risk? and Harmful Interactions: Mixing Alcohol with Medications and Drugs. Additionally, NIAAA is launching an online and print newsletter for an extramural audience. The newest issue of Alcohol Alert addresses underage drinking as a major public health challenge, and the latest copy of Alcohol Research and Health covers genetic technology in alcohol research.

• An innovative section of the Director’s Report summarizes significant papers published by NIAAA staff and grantees since the previous Council meeting to keep readers abreast of key scientific findings. A selection of 16 publications was made for this issue.

• Two of six new staff appointments were specifically noted: Dr. Steven Vogel is Tenure Track Investigator and Chief of the Section of Cellular Biophotonics in the Laboratory of Molecular Pharmacology (LMP); and Ms. Angelica Maria Rubio is working for the Office of Collaborative Research as part of the Summer Minority Internship Program coordinated by NIAAA's National Center on Minority Health Disparities.

• Four new appointments to the NIAAA Board of Scientific Counselors—the advisory body for the intramural research program—are: John Commissiong, Ph.D., Chief Scientific Officer for the Laboratory of Cell Biology and President of NeuroPharma, Inc.; Kathleen Grant, Ph.D., Professor and NIAAA investigator at the Wake Forrest University School of Medicine; Patricia Molina, M.D., Ph.D., Associate Professor in the Department of Physiology at Louisiana State University Health Sciences Center; and James Ntambi, Ph.D., Professor of Biochemistry and Nutritional Sciences at the University of Wisconsin.

At the invitation of Dr. Li, new Council member, Mrs. Hope Taft, summarized her involvement in substance abuse prevention efforts targeted at children. Her initiation came in 1986 when she was invited to a conference on crack cocaine sponsored by the Mayor of Cincinnati. At his request, she started a community coalition, Citizens Against Substance Abuse, and soon learned that alcohol is the real problem and drug of choice among young people. Mrs. Taft also helped establish Ohio Parents for Drug-Free Youth and the Ohio Alcohol and Drug Policy Alliance.

Recalling what she has learned over 25 years about prevention efforts, Mrs. Taft specified:

• The importance of forming partnerships among organizations that can not only reap monetary rewards but foster more involvement by other relevant groups.
  
  
• The requirement to use a variety of prevention strategies in multiple sectors because the many different people/groups that influence children’s lives need to reiterate the same message—that youngsters’ use of alcohol is harmful and should be socially disapproved.
  
  
• Prospects for getting parents involved increase with positive, not negative, approaches.
  
  
• The need to take a longer perspective on solving the problem of underage drinking since each new cohort of children and parents requires new education and information.
  
  
• Prevention is the key to solving substance-related problems, although efforts seem to flourish and dwindle in a cyclical flow. It is crucial to keep the message in the spotlight so the dips are smaller and the highs are higher.
  
  
• Although more attention is now given to the science of prevention, thanks largely to NIAAA, more research is still needed to ascertain what works with whom, the costs/benefits of different strategies, the effectiveness of environmental approaches–and the best combinations with individualized approaches, and the benefits of abstinence.
  
  
• More emphasis should be placed on understanding whether childhood drinking rates can be used to forecast future treatment needs—or requirements for future prisons and other services that State and Federal governments fund.

Reflecting on her role in the Leadership to Keep Children Alcohol Free, Mrs. Taft noted that this is the largest and lengthiest effort in which Governors' spouses have been involved—with 30 currently active spouses of incumbent Governors and an alumnae group of 12-15. The tenacity of the group and the consistency of its message across transitions among First Ladies is a key strength. In addition, representatives from Federal agencies and non-profit organizations who participate in the Leadership’s working committees help educate opinion leaders about adolescent development and substance abuse prevention issues. Important participants include AMA alliance members, the Surgeon General, and the spouses of Adjutant Generals in all States.

Commending NIAAA’s focus on childhood drinking issues, Mrs. Taft opined that:

• A task force similar to the one on college-age drinking would be helpful in bringing together researchers from a variety of disciplines and representatives from other sectors to focus on how to increase the age of first alcohol use and ways to help parents raise responsible, caring adults.
  
  
• The effects of the media and advertising on a child's decision-making process need further study.
  
  
• Research findings about the spike in adolescent brain development—and its ramifications —need to be more widely publicized so the public understands this crucial period is just as important as the brain development that takes place from birth to age three.
  
  
• Parents need all the support that research can provide to stay involved in the most important decision of their children’s lives—whether or not to drink!

Ms. Diane Miller, Chief of the Communications Branch, explained the background for the new public service announcement (PSA) on underage drinking prevention before showing both 60- and 30-second video versions of The Party's Over.

The PSA was developed as part of an ongoing effort to use the media for preventing or reducing early onset drinking. Several television and radio PSAs with a similar message were created several years ago. This new PSA is targeted to 8^th graders—13- and 14-year-olds. To make sure the PSA would appeal to both boys and girls as well as a broad array of ethnic groups, two rounds of focus groups were conducted in Chicago and Baltimore. The first round gathered information from 8^th graders about their attitudes and lifestyles to develop an array of messages from which two were selected. Storyboards and animatics were then created and shown to a second round of focus groups. Although the first message was dropped because it was not well received, the focus group participants found the second message relevant, sensible, and clear.

The PSA was filmed in Los Angeles, using professional actors, and produced in four lengths of 60-, 30-, 20-, and 15-seconds. This allows station managers maximum flexibility in selecting a suitable length for any available time slot. As with earlier PSAs, NIAAA expects to distribute this spot through the Office of National Drug Control Policy's media campaign that has a pro bono match and will save an estimated $8 million in advertising fees. The PSA also will be distributed to 1,000 media outlets across the country as well as to the Governors' spouses who were extremely helpful with the previous spots. Other organizations and associations that helped with previous distributions to their members will also be enlisted again.

For the future, NIAAA is developing four radio PSAs to complement this one for TV. There are also plans to create a PSA for parents, using the Surgeon General to convey the message. In concluding, Ms. Miller acknowledged the many persons who helped with the project, especially the terrific kids who participated in the focus groups.

After the two short PSAs were viewed, Ms. Miller assured Council that:

• Spanish versions were not made because Hispanic groups that were consulted indicated that Spanish-speaking youth will understand the English.
  
  
• Anything produced for parents should definitely have a Spanish version.
  
  
• The question of whether stations that usually broadcast only in Spanish will accept and air PSAs in English—even ones targeted to youth who are likely to "get" the message in either language—needs further follow-up.
  
  
• Single State Agencies will be used as a distribution resource as they were for previous PSAs. They are particularly important because they have their own media contacts.

Council member, Howard Edenberg, Ph.D., Professor of Biochemistry and Molecular Biology and of Medical and Molecular Genetics at the Indiana University School of Medicine, summarized a number of investigative approaches to the complicated question of risk for alcoholism. Because research in this area has come full circle from studies of human differences in metabolizing alcohol and risk for alcoholism to genetic components that contribute to susceptibility, he entitled his presentation A Safari: From Alcohol Dehydrogenases to the Genetics of Alcoholism and Back? The following were among the major points made:

Alcohol consumed by humans is initially converted to acetaldehyde by alcohol dehydrogenase (ADH) enzymes and then transformed to acetate by aldehyde dehydrogenase (ALDH) enzymes. While both alcohol and acetate are relatively benign substances, acetaldehyde, the intermediate product, is quite toxic in interactions with proteins, DNA, and lipids.

Seven different, but related, ADH enzymes that are encoded by seven different genes not only help metabolize beverage alcohol and other forms of alcohol, but affect risk for alcoholism and for organ damage. The ADH enzymes have evolved differences in substrate specificity and where and how they're expressed. Different coding polymorphisms— spellings of the gene that encodes the enzyme—make as much as a 40-fold difference in how rapidly alcohol is converted to acetaldehyde by that enzyme.

Assays for the known ADH enzyme variants were used to ascertain whether these differences affect risk for alcoholism among humans. Many investigators’ efforts led to the conclusion that individuals with particular variations in the alleles of the aldehyde dehydrogenase 2 (ALDH2) enzyme and the alcohol dehydrogenase 1B (ADH1B) enzymes that play pivotal roles in alcohol metabolism are less likely to become alcoholic. These "protective" alleles are particularly common among Asian populations (e.g., Chinese, Japanese, Korean), but not among Europeans. The ADH allele that’s most common among Europeans doesn't dramatically accelerate the conversion of alcohol to acetaldehyde—although there are large differences in the rates at which Europeans metabolize alcohol.

The finding that isoenzymes, and the genes encoding them, dramatically affect risk for alcoholism is one of the strongest and most widely reproduced for any complex genetic disease. The explanatory hypothesis is that high-variant alleles’ more rapid conversion of alcohol into acetaldehyde during metabolism causes a transient build-up that is probably too low to measure in blood but is, nonetheless, still important in the tissues.

To investigate the hypothesis that regulatory differences affecting the level of expression of particular ADH alleles impact risk for alcoholism and alcohol-induced organ damage, researchers scrutinized the region upstream from where transcription starts, and localized proximal promoter elements, sites where regulatory proteins bind, and other elements further upstream that make a dramatic difference in whether and where a gene is expressed in the body.

This focus on how people differ with respect to susceptibility to alcoholism and response to treatment led to an interest in the genetics of alcoholism. Many lines of evidence suggest that all diseases, including alcoholism, have a genetic component. Adoption studies have found that people are more like their biological than their adoptive parents in risk levels for alcoholism. Both twin studies and animal models have similar findings. Rats can also be selectively bred to drink alcohol for its pharmacological effects, not for taste or calories.

Risk for alcoholism, however, is a complicated issue. Although the ADH and ALDH alleles affect alcoholism risk, they don't explain all the variation. An increased susceptibility to any of the complex diseases (e.g., diabetes, hypertension, cancers, osteoporosis, psychiatric illnesses) reflects the inherited contributions of many different genes, each of which makes only a small difference and does not lead inevitably to disease. Further, genes interact not only with each other but with the external environment that includes everything from what a person consumes to the influence of social relationships and other factors. The likelihood of contracting these complex diseases is also affected by volitional choices—such as whether to drink or exercise.

To identify the specific genes in which different alleles affect risk for alcoholism and associated characteristics, NIAAA initiated the Collaborative Study on the Genetics of Alcoholism (COGA) nearly 14 years ago. This large-scale, family-based study has systematically recruited alcoholic individuals in treatment facilities at seven sites; the recruited individuals met rigorous diagnostic criteria for alcohol dependence. These subjects and all available relatives, particularly in densely-affected families, were interviewed and characterized extensively, using a specially-developed interview instrument (SSAGA) that identifies both substance use and psychiatric disorders. Electrophysiological, neuropsychological, biochemical, and genetic data were also collected.

To address the complexity and heterogeneity of both genetic and environmental factors contributing to risk for alcoholism, the sample size for the COGA project is very large. About 13,000 people have been extensively interviewed and 8,000 EEG/ERP recordings made of both probands and controls in many families. Two laboratories at Indiana and Washington Universities have generated nearly 1.25 million genotypes. The rich data set has been widely shared with genetic analysts around the world.

The strategy has been to survey the whole genome—not to guess which gene will be important. Some of the analyses have narrowed the phenotypes and looked at multiple traits together to find endophenotypes and genes that affect more than one phenotype. Microsatellite markers—the DNA regions across all chromosomes where short repeated sequences exist in many variants—have been studied to determine how alleles segregate within families. Genotyping has been used to identify rather large chromosomal regions in the genome—usually a minimum of 10 or 20 million base pairs— that show linkages with alcohol-related traits.

The linkage studies found positive evidence for regions that contain genes for phenotypes of alcohol dependence and combined alcoholic depression. These genes were discovered on several different chromosomes, but not everywhere in the genome. Chromosome 1 has a region in the middle where evidence points to a gene affecting alcohol dependence. The same region of Chromosome 1 provides even stronger evidence of a combined phenotype for either alcoholism or depression.

In the past year, COGA research has moved from identifying regions containing genes of interest to specifying individual genes that affect risk for alcoholism, using a technique called linkage disequilibrium that looks at segregation of alleles across—rather than within—families. The focus is on different markers known as single nucleotide polymorphisms (SNPs) where a single letter is spelled differently in two individuals—the very smallest variation possible. While the purpose of most SNPs is not known, some affect amino acids encoded in proteins and others affect regulatory sequences.

While most genetic studies of simple diseases (e.g., cystic fibrosis) found that a coding change in the SNP alters the function of the gene as a protein or makes a dramatic regulatory change, this model does not apply to more complex genetic diseases. Rather, SNP variations seem to induce subtle regulatory changes—not major coding differences.

To investigate the hypothesis about subtle regulatory differences, bioinformatics have been used to find candidate genes in the suspected regions. Instead of looking for rare coding SNPs, the search is directed at high allele frequencies that span the gene—whether or not they make a difference in gene functioning. This broad search is feasible because of new technology—a modified single nucleotide extension reaction mechanism and the use of mass spectrometry to determine alleles present.

Two regions on Chromosome 4 are of particular interest—one containing GABA (gamma-aminobutyric acid) genes on the short arm and the other containing alcohol dehydrogenases on the long arm. Some previous, but weak, signal evidence from the COGA studies and other NIAAA intramural efforts found risk for alcoholism in the GABA region of chromosome 4. The strong signal, using electrophysiology, was in the region where a group of four genes encode receptors for the neurotransmitter GABA. Studies of haplotypes and EEG phenotypes corroborated evidence that a specific gene encoding one of the subunits of the GABA(A) receptor has a distinct impact on risk for alcoholism. This appears to be the result of the gene’s effect on brain oscillations that modulate the overall level of neural excitation.

The strongest linkage evidence in the entire genome was a sharp peak in the same region of chromosome 4 where the ADH genes reside that related to a phenotype reflecting how much someone has ever drunk in any one 24-hour period over a lifetime. This phenotype may exert a protective influence against alcoholism by reducing tolerance to high levels of alcohol. One of the ADH genes has five markers that show evidence of association with the MAXDRINK phenotype. Thus, the safari is returning to the seven ADH genes that lie relatively close together on chromosome 4.

These findings fit the hypothesis that most of the variation that underlies complex genetic disease stems from subtle regulatory changes. In neither the GABA nor the ADH case did any of the changes that were significantly associated with alcoholism affect the amino acid encoded by the protein. This appears to confirm the hypothesis that regulatory differences, not coding differences—or in addition to coding differences—are most important risk factors as is the regulation of acetaldehyde by ADH.

New studies are also being conducted with the P and NP rats that were initially hypothesized to differ in preference for alcohol as a result of both innate differences in gene expression in their brains and different responses to the environment. Using microarray technology to look at the expression of many genes simultaneously, substantial—and promising—genetic differences between the P and NP strains have been found in the hippocampus and in ion channels and receptors.

In sum, Dr. Edenberg concluded, genes don’t cause alcoholism, but variations in genes can affect the risk for becoming alcoholic. This concept of modifiable risk needs to be communicated to the public. In the words of former NIAAA Director, Dr. Enoch Gordis, "Genes are for risk, not for destiny." As more is learned about individual differences in vulnerability and the different pathways to varied degrees of risk, non-genetic/environmental factors will also be better understood. These efforts can help reduce stigmatization and increase appreciation for alcoholism as a disease, not a moral failing. Such research can also facilitate the discovery of new pharmacotherapies for preventing or modulating the disease, targeting behavioral treatments, and encouraging recovery.

In response to questions from Council members, Dr. Edenberg clarified that:

• Twin studies have helped confirm the significant genetic component to risk, but they've not been successful at finding genes—although they might be more useful if significant genotyping were added. Since most twin studies incorporated simplifying assumptions in their statistical interpretations, case control studies are likely to be more useful for finding genes, testing their effects, and determining relative risks in the population.

• Few conclusive statements can yet be made about regulatory changes that have been found because there are too many possibilities. There could be differences in splicing patterns, in stability of RNA, or in how much RNA is transcribed.

• Evidence now suggests that one or more genes on chromosome 1 increase risk for either alcoholism or depression. The GABA genes on chromosome 4 influence alcoholism risk as do two different genes in the same cluster in the ADH region. Additional pathways to alcohol dependence are also likely and other sites are being tracked, although evidence for other receptors is less well developed.

Dr. Li, NIAAA Director and the Chair of the National Advisory Council, joined the meeting at this point, and Dr. Warren continued with the next agenda item. Referring to a document in each Council member’s folder outlining proposed changes to NIAAA’s Operating Procedures for the National Advisory Council.

The first change amends the procedures for awarding administrative supplements. Any grant supplements that exceed $50,000 in direct costs per year are shared with Council in reports provided to members at each meeting. Following a decision 2 years ago, administrative supplements under $50,000 that usually reflect minor corrections in a grant’s operations are no longer shared with Council.

The circumstances under which an administrative supplement can be awarded are listed in the formal Operating Procedures and are unchanged. These include, for example, salary increases mandated by a university or State, equipment replacement costs, serendipitous opportunities that are consistent with the specific aims of the overall project, or dissemination of specific materials. The amended procedures raise the ceiling on the amount that can be requested for an administrative supplement, a revised budget for the same fiscal year as the award, or an increase in the budget at the time of the award. Currently, the amount has been limited to no more than $100,000 per year in direct costs, but this ceiling has posed problems as the cost of grants has increased. Accordingly, the wording has been changed so that requested increases "may be made as deemed appropriate by the Director of NIAAA and generally .... should not exceed $100,000 per year or 25 percent of the direct cost, whichever is greater." All such actions are documented in official files, and supplements in excess of $50,000 are still shared with Council.

The second change pertains to expedited concurrence. Council must approve expedited concurrence parameters each year. The proposed procedures are identical to those followed over the past several years, except that the set points for applications deemed eligible for expedited concurrence are amended to lower both the percentile and priority scores. The proposed change reflects the reality that the grant process is becoming more competitive as budget limitations constrict the number of new awards. The full Council is presented principally with applications at the borderline area of funding consideration so that members can most effectively and efficiently offer decisive feedback.

The applications with the most meritorious scores are eligible for expedited concurrence. In the past, the eligibility parameters included a percentile score of 20 or better or a priority score of 200 or better—for non-percentiled proposals. The proposed change is to cut the percentile score to 15 or better and the priority score to 180 or better for non-percentiled applications. All applications with a direct cost in any year that exceeds $500,000 would automatically be excluded from expedited concurrence, as would foreign applications. NIH policy further requires that any application with a human subjects or animal welfare concern or an unacceptable code for minorities, gender, or children must undergo full Council review. An additional exclusion from the expedited process applies to all P mechanisms (e.g., program projects, centers), and U mechanisms (i.e., cooperative agreements).

For an expedited review, a Council member designated by the Chair 6- to 8-weeks before the next meeting, acts on behalf of the full Council to review a group of applications in a particular program class. All Council members are simultaneously notified that this member has been given expedited authority, and any member may recommend that any application be considered by the full Council. A 2-week delay between the call on the expedited review and the final action gives Council members an opportunity to consider the decisions. The purpose of the expediting effort is to get the best awards out earlier.

Dr. Warren explained that the Institute tries to set parameters for en bloc expediting that reflect a relation between the percentile score and the priority score for non-percentiled applications. At present, between 30 and 40 applications per round receive expedited concurrence, but this is likely to decrease in the future to between 20 and 25 applications per round.

A motion that was made and seconded to accept the proposed change in the procedures for expedited en bloc concurrence for grant applications was approved unanimously by Council.

Dr. Steven Vogel, recently appointed Acting Chief of the Section of Cellular Biophotonics in the Laboratory of Molecular Pharmacology (LMP),was formerly an Associate Professor at the Medical College of Georgia where he established a core facility for cellular imaging within the Institute of Molecular Medicine. This revolutionary new technique allows scientists to examine living cells in three dimensions. Dr. Vogel’s presentation on Imaging Cellular Logistics summarized his laboratory studies of membrane trafficking using multiphoton laser scanning microscopy and how this relates to human disease.

Using slides to illustrate his points, Dr. Vogel first distinguished various parts of a cell (e.g., plasma membrane, nuclear envelope, vesicles, organelles) and noted cells’ tendencies to move things around rather than remain in a steady state. Cells, he said, regulate when an enzyme should move into the nuclear envelope or a particular receptor should be on the cell surface. Of major interest is how cells regulate these activities, particularly for the plasma membrane, and the processes of exocytosis by which integral protein membranes get into a cell and of endocytosis by which they are removed.

NIAAA’s intramural program has a Zeiss-manufactured multiphoton laser scanning confocal microscope that provides state-of the-art capabilities for studying cellular logistics. With light microscopy, the changes in light that occur as it travels through a cell (e.g., in frequency or fluorescence, magnitude or brightness, phases) are monitored to interpret what’s happening within the cell. In fluorescence, a molecule/chromophore absorbs a specific photon that matches some change in the molecule’s energy level. When the a photon excites the molecule, internal conversions produce a power loss and a photon of a slightly longer wavelength is emitted which, in layman’s terms, is a different "color."

Over the last 20 years, scientists have discovered that many chromophores will respond to two half-energy photons in the same way as to one full-energy photon of the right power—setting the stage for two-photon microscopy that requires much higher peak power than is possible with earlier lasers, mode locked lasers have the capability for getting two photons to the same place at the same time. This is important for several reasons. While light directed on a biological sample causes damage if absorbed—limiting how long living cells can be imaged, the infrared light used in two-photon imaging is relatively benign and not absorbed by most biological samples. Further, sufficient photon density at the focal point in the two-photon mechanism produces fluorescence. Since there is no out of focus fluorescence into photon microscopy, the images are cleaner.. The duration of imaging can be increased because there’s less bleaching of the sample, and infrared light also penetrates more deeply into tissue, thus, two-photon microscopy can image deeper into living cells and tissues.

Much of Dr. Vogel’s work has focused on the membrane proteins of sea urchin eggs that are a good model for studying cells’ regulatory processes. Within a few hours after eggs are fertilized, a blastula produces apical and basal lateral specializations— sending specific proteins to specific places. If a protein receptor or G protein is destined for the plasma membrane, a vesicle has to carry it, fuse in, and deliver it. Endocytosis can reverse the process.

Images of sea urchin egg fertilization depict exocytotic activity building a barrier that prevents polyspermy—or entry by more than one sperm—and protects the embryo during early developmental stages from a potentially hostile surrounding environment. The barrier is actually formed by nearly 15,000 secretory vesicles on the egg that, upon fertilization, fuse with the membrane and drop their contents outside of the cell in an exocytosis process that can be observed in real time.

An endocytotic process can also be triggered in vitro with isolated vesicles to study what makes them respond to calcium and fuse with each other. Special dyes allow investigators to observe vesicles pinching off after endocytosis as they moved into the cytoplasm. Apparently, most cells take things back in through endocytosis soon after an endocytotic event, although it’s not clear how cells coordinate these processes.

These imaging techniques allowed Dr.Vogel’s group to discover that secretory vesicles, at least in the sea urchin egg, have a P-type, voltage-gated calcium channel that is required endocytosis . When vesicles fuse, the channel is transiently inserted into cell surface. When the cell depolarizes, the channel opens and calcium rushes in, triggering a piece of membrane to pinch off again—thereby creating a regulatory closed circuit.

With his collaborators, Dr. Paul McNeil and Dr. Kevin Campbell, Dr. Vogel recently published a landmark paper in Nature about a unique cellular defect underlying certain inherited muscular dystrophies. To investigate the hypothesis that cells—particularly mechanically challenged ones like muscles—are constantly being damaged, but have mechanisms to repair themselves, Drs. McNeil and Vogel imaged this membrane repair process, using a dye that’s only observed when it's on the membrane. When a laser punctures a muscle cell membrane, calcium rushes in and cause vesicles that might be doing other things in the cell to fuse with each other and form a patch that fills the breach. The dye doesn’t spread very much because the hole is fixed almost immediately. However, if the experiment is performed in a calcium-free saline buffer, the dye keeps on rushing in because vesicles can’t fuse with each other to form the patch.

This experiment motivated Dr. Campbell and his collaborators Iowa who were studying muscular dystrophy, to ascertain whether a similar lesion in the plasma membrane repair mechanism was involved in two forms of inherited muscular dystrophy. Using knockout mice that lacked a gene called dysferlin that's been linked to both limb-girdle muscular dystrophy and Miyoshi myopathy and control mice without this defect, Drs.Vogel, McNeill, and Campbell confirmed this hypothesis about a lesion in the membrane repair mechanism, thereby opening a new door to understanding muscular dystrophy.

Prior to these studies the concept that cells have housekeeping repair mechanisms that use membrane trafficking and endocytosis to fix holes was not appreciated. Lesions in this repair mechanism can lead to diseases. Dr. Vogel noted that one of his interests at NIAAA will be whether alcohol may also cause weaknesses in muscles that interfere with membrane repair mechanisms, causing alcohol-induced myopathy.

To conclude, Dr. Vogel described several new imaging modalities that are being developed:

• Time-resolved fluorescence—allows scientists to study biochemistry in living cells and how different proteins interact with each other by examining how long an excited molecule gives off fluorescence.
  
  
• Fluorescence resonance energy transfer (FRET)—can help determine whether two biological molecules are–or are not—interacting by monitoring the efficiency of energy transfer between them, using two fluorescent dyes.
  
  
• Correlation spectroscopy—monitors fluctuations in fluorescence created by molecules moving into and out of a small volume. Entities that are bound to each other have correlated fluctuations, whereas entities that have nothing to do with each other have completely independent fluctuations.

To a query regarding whether membrane receptors simply "wait for something to happen," Dr. Vogel responded that arguments continue about presynaptic and postsynaptic modulation of membrane receptors. In training as a neuroscientist, he favored the presynaptic modulation hypothesis and became interested in exocytosis. It appears that the numbers of receptors, as well as many other membrane proteins, can be modified. It is also unclear whether an endocytosed receptor is still active. If it is, endocytosis would appear to be a static biochemical cell state at the moment of internalization, because the extracellular milieu can't change anymore, it could potentially be a source of persisting activation—although no one has yet studied this.

Dr. Vogel also explained that the sea urchin vesicles he images are about 1 micron in diameter, compared to synaptic vesicles in a normal mammal that are about 40 nanometers in diameter.

Mr. Gregory Bloss, Public Health Analyst in NIAAA’s Division of Biometry and Epidemiology, explained the Alcohol Policy Information System (APIS)—an electronic resource that was initiated 18 months ago to help researchers understand the impact of various alcohol-related policies embodied in numerous and constantly changing statutes, regulations, and case law at all levels of government—from Federal and State through county and municipal entities.  The purpose of APIS is to encourage and facilitate research on the effects and effectiveness of alcohol-related public policies—particularly on alcohol-related health outcomes—by providing investigators with authoritative and comparable information on alcohol-related policies; by providing potential users with searchable access to this information through a public Website; and by presenting high quality information at several levels of detail ranging from summaries, tables, and charts, through the full text of laws. One notable activity undertaken by APIS is translation of the language in which laws are written to a form that is more understandable by lay persons. Unfortunately, much of the existing research on the effects of selected alcohol policies on various outcomes of interest relies on questionable data from multiple sources of varying quality and completeness.

The specific laws and policies included in APIS are currently limited to those at the State and Federal level and to enacted laws and adopted regulations—not bills that have been introduced and are still being considered by legislatures. APIS covers policies that may plausibly affect alcohol consumption, drinking behaviors, or health-related outcomes as well as those pertaining to the financing and the delivery of treatment and prevention services.

The project had a long gestation period and many contributors whom Mr. Bloss acknowledged. The 5-year contract to establish and operate APIS was awarded in September 2001 to the CDM Group in Chevy Chase, Maryland with two subcontractors in California and Minnesota. The major APIS product is a Website that’s still being developed. Information on this site is presented in two broad categories:

• The Archive of Alcohol-Related Bills and Regulations—summarizes and classifies the text of all alcohol-relevant enacted legislation and adopted regulations from State and Federal governments with links to sections of statutory and regulatory codes that are modified by the bills and regulations. The Archive can be searched by policy areas, jurisdiction, and dates.

• Detailed Information on Selected Alcohol Policy Topics—provides in-depth comparisons of State policies on selected alcohol-related topics with descriptive information, tables, maps, explanatory notes, and downloadable data as well as summaries of the current and historical status of selected policies.

Mr. Bloss demonstrated some of the main features of APIS, including the alcohol policy classification system that serves as a taxonomic framework for categorizing the bills and regulations. A sample search of policies pertaining to blood alcohol concentrations displayed an initial summary description (e.g., blood alcohol concentration limits that apply to adults, motor vehicles, boat operators, youth under age 21) with a link to relevant Federal laws, including the full text of pertinent sections. The data table had tools to control the display by dates, States, or BAC limits, with links to the statutory code sections establishing each policy. A variety of downloading options are offered so that information can be manipulated with statistical packages or spreadsheets. Another column displays the effective dates for a law (e.g., the BAC limit in Alabama was 0.08 during the period from January 1998 to January 2002).

There are plans to demonstrate APIS at scientific meetings and professional conferences, to provide technical assistance for researchers, to publish an electronic newsletter, and to compile special reports on topics of particular interest (e.g., complex policies). Over time, more topics will be added and information will be updated annually to reflect new legislation and regulations.

In response to queries, Mr. Bloss explained that:

• Expanded longitudinal coverage beyond the previous 5-years of laws and regulations will likely be limited to a few issues since retrieving information is more difficult and expensive the further back one goes. The target is to document laws that are still on the books back to 1990, but not as far back as Prohibition.
  
  
• There are no current plans for APIS to include information on licensed liquor outlets, although this might become a new topic to the extent that it is controlled by statute and regulation.
  
  
• Topics are judged worthwhile in terms of their relevance to public health, their usefulness to researchers, the feasibility of conducting the legal research, and pending changes in a particular policy area.
  
  
• APIS is designed as a research tool with authoritative and reliable information for addressing such issues as the impact of comparable policies enacted by different States and guiding policymakers’ decisions about replication of model legislative initiatives.

Dr. Alpha Brown requested the addition of information about alcohol outlets density, number, legal availability, opening times, and geographic location since these variables help shape policy and are useful for advocates who do not have the time or resources to conduct their own research.

Mrs. Taft suggested two groups that would be particularly interested in APIS: the Community Anti-Drug Coalitions of America and the National Conference of State Legislators—a group that is updating and developing a new manual to help State legislators deal with alcohol issues.

Dr. Caetano reported on a recent meeting organized by the Centers for Disease Control and Prevention (CDCP) on brief interventions in trauma centers. Attendees, including some of the leading trauma surgeons in the nation who are members of the American College of Surgeons, were interested in training colleagues to conduct alcohol screenings and brief interventions. This appears to be an important opportunity for NIAAA to develop an alcohol curriculum similar to the recent publication for primary care physicians—and a perfect occasion for translating research on brief interventions into practical applications for trauma departments. The Chairs of both the Prevention and Trauma Committees of the American College of Surgeons have indicated their willingness to support such an effort. While any curriculum for trauma surgeons would likely have to meet some type of accreditation standards, they could help disseminate the information.

In response to a query about policies or laws pertaining to payments for trauma surgeons who identify alcohol-related problems, Dr. Caetano agreed that some legislative changes may be needed on a State by State basis in order for trauma surgeons to be able to identify alcohol related injuries.

Dr. Karen Peterson, Office of Scientific Affairs, spoke about the Advanced Alcohol Research Program (AARP) that NIAAA initiated in 2002 to undertake high-risk, potentially high-impact research. This project, modeled on the Defense Advanced Research Projects Agency (DARPA), reflects a paradigm shift in scientific approach by accelerating/compressing the time frame for research rather than following incremental, carefully constructed, traditional methods. The NIH Roadmap Committee for High-Risk Research is considering whether a DARPA-type approach should be incorporated throughout NIH.

The DARPA method gives Program Managers a more critical role in research activities than allocated to traditional Program Officers. They have more decision-making responsibilities, fewer constraints on their time and resources, and more direct contact with contractors. NIAAA has been fortunate in hiring two former DARPA Program Managers for its program: Dr. Dennis Healy, a Professor of Mathematics at the University of Maryland, is in charge of the Biosensors Program, and Dr. Anna Tsao, from AlgoTech, is directing the Ecosystems Modeling Program.

At present, NIAAA has three AARP projects: 1) the biosensor and data sensing systems effort has commitments of $3 million for FY 2003 and FY 2004; 2) the ecosystem modeling of alcohol-related behavior project has a FY 2004 commitment of $1 million in 2-year seed monies; and 3) a computational neuroscience and alcoholism project is still being developed.

For the Biosensor project, a Request for Proposals (RFP) calling for 2-year contracts with options for 3 additional years was released in 2002 and awards made that fall. Two contracts went to Indiana and Duke Universities, and three additional awards in 2003 were made to Brown University, SpectRx, and Science and Engineering Systems Incorporated (SESI). At the same time the RFP was released, NIAAA issued a Small Business Innovation Research (SBIR) RFA for a similar biosensor product and made four SBIR Phase I grant awards that are each for a 2-year period instead of the usual 6 months.

The RFP for the biosensors project required products to meet several stringent criteria, including:

• Links between an alcohol-sensoring device and a data-sensing system so that large quantities of reliable data could be collected, stored, and rapidly analyzed in real-time.
  
  
• Continuous measurements of alcohol concentrations at least every 1- to 5-minutes.
  
  
• Being comfortably wearable by the person (or animal) being monitored.
  
  
• Sufficient storage and/or telemetric data transmission capabilities for at least a month of continuous activity so that wearers are not burdened unreasonably by downloading needs.
  
  
• Capability, at minimum, to measure alcohol levels in blood, but ideally able to monitor alcohol in the breath and in other bodily fluids and organs—and also to integrate these data with other physiologic measurements such as heart rate, blood pressure, temperature, and other metabolites of ethanol such as acetaldehyde.
  
  
• Capacity to detect alcohol within a broad range of 5 to 500 milligrams per deciliter so that the imbiber as well as the binge drinker can be monitored.
  
  
• Being relatively tamper-proof and accurate through a range of normal physical activities.

Dr. Peterson summarized the five contractors’ plans and progress in developing these biosensors:

• Indiana University—as part of a consortium with the University of Tennessee, Knoxville, and Purdue University—is making an implantable sensoring device that uses piezo-resistive microcantilevers whose vibration frequency changes when alcohol binds to their polymeric coatings. The small, capsule-like device, which has some potential for looking at other analytes such as acetaldehyde, is fingertip size and will be implanted behind the armpit, just below the shoulder. The device contains a microelectrical-mechanical systems (MEMS) transducer that is about the diameter of a human hair, an analog-to-digital converter with the capability of making one-second queries every 16 minutes for a week, a memory source, a timer, a transmitter, a link to the physiologically-based pharmacokinetic modeling system, and a too-small battery power source whose capabilities the contractors would like to expand.

• The group from Duke University Medical Center and Engineering School and the Georgia Institute of Technology is making a compact, non-invasive biosensor that will use near-infrared Raman spectroscopy and communicate telemetrically with a data station. These engineers hope to construct three different sized instruments. The spectroscopic technique will allow measurement of multiple analytes. The unique component is a multimodal multiplex spectroscopy (MMS) that relies on lasers that diffuse on a person’s finger—which is inserted into the device— to measure alcohol levels that the detector interprets. Instead of getting a single, small spectrum, the device collects a million spectra simultaneously and interprets these mathematically. Different peaks in the spectra represent different concentrations of alcohol—from pure ethanol to 1 percent ethanol.

• The Brown University group, working with an SBIR grantee, is using a wristwatch-like transdermal sensor recorder with a platform developed by Giner. The plan is to develop an optical sensing system to replace the electrochemical system now used. Additionally, the device will have a full-body sensor that can derive ethanol ingestion and liver and brain alcohol levels from a monitor signal.

• SpectRx has already patented a minimally-invasive device for extracting, measuring, and analyzing the interstitial fluid that lies just below the top skin layer and contains many analytes as well as glucose. This sensor combines physiologic, environmental, and direct alcohol sensing and will be capable of analyzing many different analytes simultaneously to produce an individualized alcohol profile for each wearer. The collaborators are Children's Hospital in Los Angeles and Honeywell Corporation.

• Science and Engineering Services, Inc., (SESI) is developing a non-invasive device that uses near-infrared absorption—rather than Raman—spectroscopy to examine a combination of wavelengths. The frequency of the laser will be pseudo-randomly modulated to produce a much better signal resolution. The wearable sensing monitor will examine both breath and blood alcohol levels—with a capability to detect alcohol in 1- to 5-seconds at 0.005 percent blood alcohol concentrations.

Turning to the ecosystems modeling program, Dr. Peterson explained that $1 million in 2-year seed monies for FY 2004 will be used to analyze interactions between drinking and the community. The interest in ecosystems modeling was stimulated by three NIAAA-sponsored workshops last year in New Orleans, Philadelphia, and Berkeley. A broad RFP for this effort will soon be released, with awards expected in early 2004 to one or two projects that will test the feasibility of this innovative approach. Contract awardees will be expected to develop dynamic systems models of alcohol-related behaviors in populations—a very different concept than static models. Successful projects will demonstrate a strong potential for a priori analysis of success, failure, unknowns, and costs of potential community prevention and intervention strategies. Applicants will select a particular population (e.g., college, airbase), setting (i.e., urban, rural) and data set.

In response to questions, Dr. Peterson clarified that:

• Measurements by the proposed biosensors should not be impacted by environmental factors and other bodily functions (e.g., temperature, sweat). While perspiration is an issue for the Brown University project, the strong seal on the wristwatch that Giner is developing has successfully resisted disruption. SpectRx has been testing the glucose monitor for several years without significant problems, and the other devices are not as vulnerable to the noted environmental problems.

• The biosensor devices approximate alcohol levels in venous, not arterial, blood. Although there are advantages to measuring alcohol in various places, some gold standard must be accepted eventually. While venous blood is usually proposed as the model, breath measures most accurately reflect what’s happening in the brain and explain NIAAA’s interest in measuring both blood and breath levels of alcohol.

• Two of the projects are experimenting with whole-body or different-organ models. Since alcohol levels are different in many parts of the body, the Indiana University group is developing a method to extrapolate findings from one part of the body to another.

Dr. Taylor offered to forward recent articles about new techniques to measure antibiotic levels in interstitial fluid that can be used to calculate dosing requirements. Dr. Hoffman commented that NIAAA might want to leverage DARPA’s previous bio-surveillance work under two initiatives (i.e., Essence, Leaders) because it seems similar to the proposed ecosystems modeling project.

Dr. Paul Gruenewald, Scientific Director and Senior Research Scientist at the Prevention Research Center (PRC) in Berkeley, California as well as Principal Investigator for two environmentally-focused, NIAAA-funded grants, spoke about Population Models of Alcohol Problems. The following were among the salient points:

Studies of the growth and decline of human populations incorporate demographic, economic, sociological, and geographic theories into examinations, for example, of disease processes or different types of morbidity and mortality. Some of the human population models are codified in community systems theory, exchange theory, and economic geography.

The types of alcohol-related problems addressed in population models are usually such acute effects as alcohol-related crime, injuries, death, auto crashes and fatalities, or lost productivity. These problems change rapidly over time, reflect interactions of individuals with their environments, and are sufficiently complex to be mathematically interesting.

Three issues of major current interest in alcohol-related population models pertain to: 1) the distribution of alcohol-related problems in the population; 2) how facets of the alcohol environment contribute to alcohol-related problems in populations; and 3) how geography influences alcohol-related problems.

The distribution of alcohol-related problems in the population:

• Knowing what segments of the drinking population produce the most problems, we can efficiently target environmental prevention programs (e.g., at specific at-risk groups or at the broader population of users).
  
  
• New mathematical models of drinking patterns enable us to use survey data to (a) calculate the distribution of drinking levels among alcohol users, (b) assess dose-response functions relating drinking levels to problem outcomes, and (c) examine the population distribution of drinking problems.
  
  
• This research suggests that problems are broadly distributed across drinkers and drinking levels, supporting the use of broad-based environmental preventive interventions.
  
  
• Applications of these models also show that incidents of very extreme levels of drinking, a particular problem among college students, can be predicted using population models. In any 30-day period, 5% of drinking events among college males will be ones in which consumption exceeds 24 drinks.