39th Annual John B. Graham Research Abstracts

 

Locally administered antibiotics for prophylaxis against surgical wound infection: An in vivo study

 

Seth R. Yarboro, BS, Elyse J. Baum, BS, Laurence E. Dahners, MD

 
Department of Orthopaedics
University of North Carolina, Chapel Hill, NC
 
 

This study investigated the relative efficacy of various methods of antibiotic delivery for the prevention of surgical wound infections. Currently, the standard of surgical infection prophylaxis consists of perioperative systemic antibiotics. We hypothesized that sustained release of local antibiotics inside the wound cavity by a drug delivery system (DDS) would be more effective than systemically administered antibiotics. Using a rat model, we inoculated a surgical wound in the quadriceps musculature with 8.0 x 105 CFU of Staphylococcus aureus (ATCC 29523), then administered one of seven treatments. There were 10 rats in each of the groups, consisting of: control (no treatment), bacitracin irrigation, calcium sulfate flakes, systemic gentamicin, local aqueous gentamicin, local gentamicin loaded calcium sulfate flakes, and a combination of local gentamicin loaded calcium sulfate and systemic gentamicin. To further evaluate a trend, the systemic gentamicin and local gentamicin solution groups were extended to include 25 and 27 rats, respectively. At 48 hours post surgery, the wounds were quantitatively cultured. The control, bacitracin irrigation and plain calcium sulfate groups had very high bacterial counts and high mortality rates while the gentamicin treatment groups had low bacterial counts and full survivorship. Local gentamicin was significantly more effective than systemic gentamicin. Refuting our hypothesis, the gentamicin loaded calcium sulfate flakes did not result in significantly lower bacteria counts than the systemic gentamicin. However, simple local gentamicin solution injected directly into the closed wound did result in significantly lower levels of bacteria than the systemic gentamicin. We believe that a high initial concentration of the locally applied antibiotic inside the wound effectively kills bacteria present in the wound cavity where systemic antibiotics have poor penetration, suggesting that this method of antibiotic administration may be a desirable adjunct for the prophylaxis of infection in surgical wounds.

 
 

Evolution of the Surgical Management of Neonatal Ovarian Cysts: Laparoscopic-Assisted Trans-umbilical Extra-corporeal Cystectomy (LATEC)

Lucy Schenkman1, J.Duncan Phillips1, Timothy Weiner1

1Department of Surgery

University of North Carolina, Chapel Hill, NC

 
 

 Background: Since its first detailed description in 1995, laparoscopic management of neonatal ovarian cysts has typically required multiple incisions, specialized equipment, and advanced laparoscopic skills. After some initial frustration with neonatal laparoscopy, we developed a simplified single-incision, Laparoscopic-Assisted, Trans-umbilical, Extra-corporeal Cystectomy (LATEC).

Objectives: This study reviews our experience with this technique and compares outcomes to those of our other surgically-managed neonatal ovarian cyst patients.
Methods: Retrospective record review of 20 patients treated surgically between 1992 and 2006. Student's t-tests and ANOVA were used for comparisons (p<0.05=significant).
Results: Means: age 11 dys, weight 3.7 kg, cyst diameter 5.0 cm. 19 patients were diagnosed prenatally, at mean gestational age 33 wks. 12 of 20 (60%) had torsed cysts (1 bilateral). 3 of 13 torsed cysts (23%) were less than 4 cm diameter (range 2.9-3.5 cm). Laparotomies were transverse lower abdominal incisions. Laparoscopic operations used 2 incisions (3 patients) or 3 incisions (2 patients). LATEC involved trans-umbilical laparoscopy, complete cyst aspiration, and then cyst evisceration through the umbilicus for either ovarian cystectomy (simple cysts) or salpingo-oophorectomy (torsed cysts). Laparoscopic patients had similar time to feeds, length of stay (LOS), and postop narcotic requirements when compared to laparotomy patients (p=NS) but a high complication rate. LATEC patients had shorter surgical times (45.6 min versus 83.0 min)*, more rapid advancement to full enteral feedings (20.5 hrs versus 38.7 hrs)*, and shorter lengths of hospital stay (30.9 hrs versus 53.3 hrs)* when compared to traditional laparoscopy (*p<0.05). LATEC had equal ovarian preservation (100% of ovaries with simple cysts) to traditional laparoscopy. There were no major complications in the LATEC patients, and cosmetic results with LATEC were excellent.
Conclusions: Both laparoscopic and "open" approaches have low morbidity and rapid recovery. LATEC is a relatively simple procedure which combines laparoscopy and traditional extra-corporeal surgery and appears to offer improved outcomes, with a single incision.

 
 
 

Role of Myosin-X in the Formation and Function of Microvilli

Katy Liu, Damon Jacobs, Richard Cheney

Department of Cell and Molecular Physiology, UNC-Chapel Hill

 

            Myosin-X is an actin-based motor involved in intercellular transport. It has recently been shown that myosin-X is essential for filopodial formation and localizes to the tips of filopodia while exhibiting bidirectional movement along its length. This study investigates the role of myosin-X in a similar construct, microvilli. Microvilli are slender extensions composed of a core of actin filaments which is responsible for absorption and secretion by intestinal epithelial cells in the small intestine. As myosin-X is plays a necessary role in filopodia, we question whether myosin-X behaves similarly in microvilli, specifically, in its localization and its function in the formation and maintenance of the microvilli.

            We utilized the Caco-2 construct, a human colon carcinoma cell line which most closely resembles human intestinal epithelial cells in structure and function. Localization of endogenous myosin-X was determined by epifluorescence staining over a two-week time course and viewed with confocal microscopy. Also, the quantity of myosin-X over the time course was determined by western blot. Over-expression of myosin-X was carried out through transient transfection of the Caco-2 cells with GFP-labeled myosin-X and was viewed with confocal microscopy.

            Analysis of endogenous myosin-X in Caco-2 cells showed that myosin-X in cells in the non-polarized state (0-5 days) localizes to the tips of filopodia-like villous structures (immature microvilli). As the cells mature and became polarized (5-13 days), myosin-X then localizes all along the microvilli and concentrates in vesicles within the cell cytoplasm. Over-expression of myosin-X in mature Caco-2 cells shows decreased polarization and a less developed brush border. This suggests that myosin-X plays a role in the formation of microvilli at the filopodia-like stage (0-5 days) and in maintenance of polarization of microvilli in the mature stage (5-13 days). Further data will be elucidated through viral knock-down and expression of myosin-X in Caco-2 cells.

 
 

Anti-estrogen mediated enhancement of ATP-mediated ASL secretion in cystic fibrosis

 Russell Stackhouse, Robert Tarran1, and Mark Clunes

1Department of Cellular and Molecular Physiology

University of North Carolina, Chapel Hill, NC
 
 

Cystic fibrosis airways exhibit a decreased airway surface liquid (ASL) height due to a lack of fluid secretion. This results in reduced mucus clearance and an impaired innate immunity leading to bacterial infections and destruction of the airways. It has been shown that the estrogen receptor antagonist tamoxifen (TXN) can potentiate ATP-mediated ASL secretion, which is signaled via changes in intracellular calcium, which may be therapeutically beneficial for CF patients. Our aim was to evaluate whether other anti-estrogens were more efficacious than tamoxifen in stimulating increases in intracellular Ca2+. JME cells, derived from a CF patient, were stained for intracellular Ca2+ and measured by epifluorescent microscopy. TXN, ICI 182,780 (a pure anti-estrogen), and 4-hydroxytamoixfen (4-HT; the major metabolite of TXN) were applied to the cells in the presence of ATP to create dose response curves. BHK cells, that do not express  or  isoforms of the estrogen receptor, were also treated with TXN. TXN showed significant potentiation of the calcium response to ATP in cells that express estrogen receptors. Neither ICI182,780 nor 4-HT showed potentiation of the calcium response in these same cells. When tamoxifen was added to BHK cells that do not exhibit estrogen receptors  or  the same level of potentiation of the calcium response to ATP was witnessed. The lack of potentiation by ICI182,780 and 4-HT indicates that TXN’s potentiation of the Ca2+ response to ATP is mediated in some other way than the classical ER.   The same response seen in the BHK cells that do not exhibit the ER further supports this conclusion. Further research should be performed to better elicit TXN’s mode of potentiation on the intracellular Ca2+ response to ATP.

 
 
 

Proyecto Puentes de Salud (Project Health Bridges): The relationship of economic migration and HIV risk factors in Rural Mexico.

Ian J Nelligan1,2, Amanda L Rollins1,2, Allan D Nanney1,2,  Trista D Snyder1,2, Pamela Y Frasier MA, MSPH, PhD1, Alfred Reid MS1, Bron Skinner PhD1, Daniel Reuland MD2, Mauricio G Cohen MD3

1Department of Family Medicine, Department of Internal Medicine2, Department of Cardiology3

 

Several well recognized studies have defined risk factors for HIV transmission among male economic migrants in NC and the US, however, there remains a deficit in primary research of HIV risk among the partners of economic migrants in rural Mexico. A convenience sample of 169 participants, recruited from six rural communities in Guanajuato, Mexico were offered free HIV testing using the Oraquick® Advance™ Rapid HIV-1/2 Antibody Test and counseled about risk factors and reduction strategies for HIV by bilingual investigators. A counselor-conducted survey was used to assess extent economic migration impacted the rick of acquiring HIV. Additionally, self-administered HIV knowledge survey was used to assess the basic and advanced knowledge of HIV and risk reduction. Women comprised 85.8% of those tested. Aproximately 90% of participants neither migrated nor used condoms routinely; however, those who used condoms regularly were 4.6 times more likely to have migrated. Those who migrated were 48 times more likely to have sex with a commercial sex worker and 67% never or almost never used condoms with them. Risk of HIV transmission is high in this population while condom was very low. Those that migrated to the US were much more likely to engage in high risk behavior. Participants that never left Mexico have risk derived from transnational migration between communities with a steep gradient of HIV incidence.

 
 
 
 
Synopsis:
 

Introduction: A collaborative Duke/UNC study in Durham, NC revealed that 46% of single Hispanic men, and 42% men living apart from their wives, had intercourse with sex workers within the last year. While increased risk factors for HIV transmission among male economic migrants in NC and the US have been established in several studies, there is a lack of information about HIV risk among the spouses of economic migrants in rural Mexico. Using a preliminary HIV risk assessment survey, we selected six rural communities near Juventino Rosas (JR), Guanajuato, Mexico with estimated 66% or more of the working men migrating to the US and a large number coming to the South. JR was chosen because of the Sister City and Sister Church relationships with Carrboro, NC and the Catholic Church of Chapel Hill, NC. Identical screenings are planned in Orange and Chatham Counties, NC.

 

Methods: A convenience sample of 169 participants, recruited from six rural communities in Guanajuato, Mexico during June and July of 2006, were (1) offered free HIV testing using the Oraquick® Advance™ Rapid HIV-1/2 Antibody Test and (2) counseled about risk factors and reduction for HIV by bilingual investigators. The participant was then offered the free, confidential HIV screening. All participated in a counselor-conducted survey to assess economic migration defined as “migration to the US to live or work for 1 month or more” and risk of acquiring HIV. A self-administered HIV knowledge survey was used to assess the basic and advanced knowledge of HIV and risk reduction of 34 individuals in one community. Networks were set up with local doctors to provide care in positive test situations.

 

Results: Using Oraquick® Advance™ Rapid HIV-1/2 Antibody Test, 169 people were tested for HIV, 0.0% were positive. 169 individuals participated in our study. Women comprised 85.8% of those tested and 11% of those sampled had been to the US.

Analysis of the HIV risk survey showed that aproximately 90% of participants neither migrated nor used condoms routinely; however, those who used condoms regularly were 4.6 times more likely to have migrated. Of those screened, participants who migrated were 17 times more likely to have had more than one sexual partner. Those who migrated were 48 times more likely to have sex with a commercial sex worker and 67% never or almost never used condoms with them. 61% of those that have migrated had sex with a commercial sex worker compared with 3% that had not migrated. 

 

Discussion: While incidence of HIV was not found, the risk of HIV transmission is high in this population. Condom use in this population was very low. Those that migrated to the US were much more likely to engage in high risk behavior including multiple sexual partners, little condom use and sex with commercial sex workers. Those that never left Mexico had little risk as a group but many had one life time partner that introduced risk. The investigation was limited by being a convenience sample of those attracted to a health fair setting and the under-representation of male participants due to the fact that the vast majority was in the US at the time. HIV knowledge was very limited. Data analysis is ongoing. 

 
 

Impact of a 12 Week School-Based Intervention on

Nutrition and Physical Activity Knowledge, Attitudes, and Behaviors

 in Rural Teens and Elementary School Children:

Improving Meals and Physical Activity in Children and Teens (IMPACT)

Natalie Muth1, Avik Chatterjee1, Donna Williams3, Kori Flower2

1School of Medicine and 2Department of Pediatrics, University of North Carolina, Chapel Hill, NC;3Orange County Schools, North Carolina

 

Poor nutrition and inactivity are widespread and contribute to the epidemic problem of childhood overweight. This study examines the effectiveness of a pilot program to improve nutrition and activity in 4th graders and teens in a rural North Carolina community. The Improving Meals and Physical Activity in Children and Teens (IMPACT) school-based curriculum used a train-the-trainer model to encourage increased activity and healthier nutrition. Six medical students taught nine teens (ages 16-17) recruited from one high school the IMPACT curriculum and leadership skills during a 15-hour training program. Four 4th grade classes at a neighboring school were randomized to receive the IMPACT curriculum delivered by the medical students and teens over 12 weeks (2 classrooms, 38 students) versus standard curriculum (2 classrooms, 37 students). Outcomes were assessed using pre-intervention and follow-up survey and BMI measures.  The hypothesis is the teens and 4th grade intervention groups will increase (1) nutrition and physical activity knowledge and attitudes; (2) daily fruit, vegetable, whole grain, and dairy product consumption; and (3) amount of physical activity. Results indicate that 4th graders in the intervention group significantly increased fruit and vegetable intake (p<0.05), knowledge of food group to eat the most servings (p<0.01), and recommended number of servings of fruits and vegetables to eat daily (p<0.01). The number of teens lost to follow up prohibited analysis of teen outcomes. If disseminated more widely, the IMPACT curriculum could support healthy nutrition and activity in high school and elementary school settings.

 
 
 

Bradykinin B1 and B2 receptors both have protective roles in renal ischemia-reperfusion injury

Robert W. McGarrah, Masao Kakoki, Hyung-Suk Kim and Oliver Smithies

Department of Pathology and Laboratory Medicine

University of North Carolina, Chapel Hill, NC
 
 

Ischemic events such as stroke, myocardial infarction and acute renal failure are well-known and devastating complications in patients with hypertension, diabetes and atherosclerosis. Moreover, damage incurred during ischemia/reperfusion (I/R) of transplanted allografts directly impacts short-term and long-term graft outcomes. Angiotensin-I converting enzyme inhibitors (ACEIs), in addition to their beneficial effect on chronic cardiac remodeling after ischemia, have been demonstrated to prevent acute injury caused by I/R in the heart, lung, liver and kidneys. Studies indicate that this protective effect is due mainly to activation of the bradykinin-nitric oxide (NO) cascade rather than suppression of angiotensin II formation. Indeed, angiotensin receptor blockers are much less effective than ACEIs in protecting against I/R injury and bradykinin receptor antagonists and NO synthase inhibitors attenuate the protective effects of ACEIs. There are conflicting results, however, regarding the exact role of bradykinin receptor stimulation in I/R injury and some studies suggest that stimulation of the bradykinin B1 and bradykinin B2 receptors may have opposite effects from each other in this situation.

We used mice lacking either the bradykinin B2 receptor gene (Bdkrb2-/-) or both the bradykinin B1 and B2 receptor genes (Bdkrb1-/-Bdkrb2-/-) to further investigate the role of the bradykinin receptors in renal I/R injury. After 30-minutes of bilateral renal occlusion and 24-hours of reperfusion, mortality rates, renal histological and functional changes, 8-OHdG levels in total DNA (a marker of oxidative DNA modification), mitochondrial DNA deletions, the number of TUNEL-positive cells, and mRNA levels of transforming growth factor β1 and endothelin-1 in the kidneys of Bdkrb1-/-Bdkrb2-/- mice were significantly more enhanced than in Bdkrb2-/- or wildtype mice. Therefore, both the bradykinin B1 and B2 receptors play an important protective role in reducing DNA damage, apoptosis, morphological and functional kidney changes, and mortality during renal I/R injury.

 
 
 
 
A Health and Nutrition Needs Assessment

of Six Rural Honduran Communities Served

by the UNC School of Medicine Honduran Health Alliance

Natalie Digate Muth, MPH, RD1, Alice Ammerman, DrPH, RD2
 

1Department of Family Medicine, 2Department of Nutrition

University of North Carolina, Chapel Hill

 

Honduras is one of the poorest Latin American countries with over 68% of its residents classified as at or below the poverty level. Honduras also suffers from increased rates of insufficient food intake, protein malnutrition, low birth weight, and stunted growth. The situation is even worse in rural communities. The UNC Honduran Health Alliance (HHA) exists in an effort to improve the health status of rural Honduran families. The purpose of this study was to determine the target population’s health and nutrition status based on a variety of factors including background conditions; community conditions; living, working and social conditions; and individual lifestyle factors. The community needs assessment was conducted in four stages: a literature review, a focus group meeting with four of the Honduran lay health advisors, focus groups with community members who attend the educational Charlas (focused health discussions led by the Honduran Health Alliance medical students), and 24 hour dietary recall and BMI measurements. Results indicate that women studied maintain a healthy weight, eat an unvaried diet low in fruits and vegetables, and have a stronger desire to eat more fresh produce and a more varied diet but feel they have limited access to those foods. The HHA is in a unique position to help community members develop creative solutions to improve dietary intake including modifying educational teaching and collaborating with community members on creating sustainable projects such as a community garden.

 
 

Clinical Characteristics and Prognostic Factors of

Stenotrophomonas maltophilia Infections

Lauren Paton1; Jonathan R. Salahshour1,2; Aram Kim1,2 and Craig Barrett1, Toan T. Huynh1; Britt Christmas1, Ronald Sing1

1Department of General Surgery

Carolinas Medical Center, Charlotte, NC

2School of Medicine

University of North Carolina, Chapel Hill, NC

 

Appropriate empiric antimicrobial therapies against life-threatening bacterial infections such as meningitis, pneumonia, and sepsis has been shown to decrease morbidity and mortality as compared to therapies strictly based on a definitive culture diagnosis. The decision to cover select virulent and/or common organisms is a dynamic process that involves re-evaluation of the patient population and the infectious organisms responsible. Stenotrophomonas maltophilia is a multidrug-resistant, gram-negative rod which causes nosocomial infections in immunocompromised patients and patients who are subjected to invasive devices such as catheters or endotracheal tube. Currently, Stenotrophomonas maltophilia infections are not included in empiric treatment coverage for pneumonia and sepsis for the patient population susceptible. To analyze the effects of empiric therapy on the clinical outcome of Stenotrophomonas maltophilia infections, all surgical patients at the Carolinas Medical Center from May 2003 to July 2005 treated for Stenotrophomonas maltophilia infections were retrospectively reviewed. Patient demographics, length of stay (LOS), concomitant infections, antibiotic susceptibilities and mortality was analyzed with standard statistical testing with a confidence interval of p<0.05. The study identified eighty-two patients (37 females/ 45 males) with an mean age of 54.6 years (range; 12-92). Mean LOS was 38.1 days (range; 2-120) with a mean ICU-LOS 29.3 days (range; 0-114 days). Overall mortality was 26.8%. The most common Stenotrophomonas maltophilia infections were pneumonia (68%) and bacteremia (11%). Thirty-three patients (40%) had a past medical history significant for COPD or asthma. The diagnosis of Stenotrophomonas maltophilia was made on average hospital day number 17 (range; 0-86). Forty-seven patients (57%) had received antibiotics for other infectious etiologies prior to Stenotrophomonas maltophilia infection. Fifty-nine patients (72%) had empiric treatment that required a change in antibiotic therapy based on sensitivity results. Patients requiring antibiotic changes based on sensitivities did not have worse outcomes (p=0.64) however those with the diagnosis made early in their hospital course within 2 weeks (p=0.05) had significantly better outcomes. Though Stenotrophomonas maltophilia infections seemed to occur in patients treated for prior infections including ventilator associated pneumonia, empiric treatment against Stenotrophomonas maltophilia did not correlate with improved clinical outcomes. At this point, we do not recommend that Stenotrophomonas maltophilia be included in empiric antibiotic coverage.

 
 

A Comparison of Fixation Methods for Acromioclavicular Joint Separation: A Biomechanical Study

Michael Hromadka, BS 1, Laurence Dahners, MD2,3, Paul Weinhold, PhD3

1School of Medicine, 2Department of Orthopaedics, and 3 Department of Biomedical Engineering University of North Carolina, Chapel Hill, NC

 

Acromioclavicular (AC) joint dislocations and distal clavicle fractures can be treated in a variety of ways including benign neglect, brace stabilization, or surgical intervention. Surgical stabilization of the coracoclavicular interval is problematic with no obviously superior method available; however, a percutaneous method would be desirable. In a controlled laboratory study we compared the biomechanical characteristics of the traditional Bosworth screw with a suture anchor technique in order to determine if they have comparable strength and stiffness. The suture anchor technique uses two suture anchors that can be placed percutaneously into the coracoid and the sutures are then tied together over the clavicle. Our results showed that the Bosworth screw had a mean maximum load at failure of 903(SD=72) N and a stiffness of 200(SD=77) N/mm. The suture anchor technique had a mean maximum load of 800(SD=182) N and a stiffness of 76(SD=26) N/mm. The difference in the load at failure was not statistically significantly different but the stiffness of the Bosworth screw was statistically greater, p-value =0.036. The suture anchor results are not dissimilar to the load to failure of 725 (SD=230) and stiffness of 115.9 (SD =36.2) previously reported by Motemedi upon testing of intact cadaveric coracoclavicular intervals. The double suture anchor technique appears to provide sufficient strength to adequately stabilize the coracoclavicular interval and provides stiffness similar to that of the intact coracoclavicular ligament system. We believe that these data suggest a clinical trial may be in order to evaluate the use of the double suture anchor technique for the stabilization of the coracoclavicular interval.

 
 

Proyecto Puentes de Salud (Project Health Bridges): The relationship of economic migration and HIV risk factors in Rural Mexico.

Amanda L Rollins1,2, Ian J Nelligan1,2, Allan D Nanney1,2, Trista D Snyder1,2, Pamela Y Frasier MA, MSPH, PhD1, Alfred Reid MS1, Bron Skinner PhD1, Daniel Reuland MD2, Mauricio G Cohen MD3

1Department of Family Medicine, Department of Internal Medicine2, Department of Cardiology3

University of North Carolina, Chapel Hill, NC
 

While increased risk factors for HIV transmission among male economic migrants in NC and the US have been established in several studies, there is a lack of information about HIV risk among the spouses of economic migrants in rural Mexico. A convenience sample of 169 participants, recruited from six rural communities in Guanajuato, Mexico were offered free HIV testing using the Oraquick® Advance™ Rapid HIV-1/2 Antibody Test and counseled about risk factors and reduction for HIV by bilingual investigators. All participated in a counselor-conducted survey to assess economic migration defined as “migration to the US to live or work for 1 month or more” and risk of acquiring HIV. A self-administered HIV knowledge survey was used to assess the basic and advanced knowledge of HIV and risk reduction to some participants. 0.0% were positive. Women comprised 85.8% of those tested. Aproximately 90% of participants neither migrated nor used condoms routinely; however, those who used condoms regularly were 4.6 times more likely to have migrated. Those who migrated were 48 times more likely to have sex with a commercial sex worker and 67% never or almost never used condoms with them. Risk of HIV transmission is high in this population. Condom use in this population was very low. Those that migrated to the US were much more likely to engage in high risk behavior. Those that never left Mexico had little risk as a group but many had sexual partners that introduced risk. Overall, HIV knowledge was very limited.

 
 
 
 
 
 
 
 
 

     The effects of recombinant Fc (rFc) on passive transfer mouse models of bullous pemphigoid, pemphigus foliaceous and pemphigus vulgaris

 
Gene Bain
Advisor: Dr. Zhi Liu
 

     Bullous pemphigoid (BP), pemphigus foliaceus (PF) and pemphigus vulgaris (PV) are autoimmune skin blistering diseases, each with its own characteristic pathophysiology. Pemphigoid exhibits subepidermal blisters, infiltration by inflammatory cells as well as an accumulation of IgG autoantibodies at the basement membrane zone (1). Pemphigus, on the other hand, progresses through intraepidermal blisters as a result of epidermis specific antibodies (2). One form of treatment that has shown considerable promise for patients with other autoimmune disorders has been intravenous IgG (IVIG) (3). However, IVIG as a therapy for the above blistering diseases has yet to be fully evaluated. Because of its effectiveness in reducing the manifestations of BP, PF and PV in mice models, more work is being done with respect to this therapy’s mechanism of action. Based on recent work in this area, this study seeks to examine the efficacy of a recombinant protein that makes use of a specific domain within the IgG molecule-the Fc domain. It is our hope that through the use of this specific domain of the immunoglobulin, an efficacy equivalent to that of IVIG in terms of disease reduction will be achieved as demonstrated through these same in vivo models. 

 
 

Screen for miRNAs that Regulate Apoptosis in Primary Neurons

Adam J Kole1, Michelle I Smith2 and Mohanish Deshmukh2

1MD/PhD Program, 2Department of Cell and Developmental Biology

University of North Carolina, Chapel Hill, NC
 
 

Apoptosis, or programmed cell death, is an evolutionarily conserved process by which cells die in a regulated manner. Apoptosis is important both in development and in homeostasis by removing cells that are superfluous, no longer useful, or harmful for the organism. The deregulation of cell death has many pathological consequences. MicroRNAs (miRNAs) are a newly discovered family of evolutionarily conserved regulatory RNAs. miRNAs are short, non-coding RNAs that are processed into double-stranded RNA structures and regulate gene expression by binding to complementary mRNAs, inhibiting their translation. Several miRNAs have been identified in Drosophila that are potent inhibitors of cell death by blocking translation of apoptotic proteins.

 

These findings prompted us to investigate whether expression of miRNAs is modulated in mammalian primary neurons undergoing apoptosis. Since miRNAs are involved in the regulation of expression of a wide range of genes, it was hypothesized that miRNAs do in fact regulate neuronal apoptosis. The specific aims to test this hypothesis were:

          To culture murine primary cerebellar granule neurons (CGNs).

          To induce apoptosis in primary CGNs.

          To purify RNA from CGNs and confirm the activation of apoptosis by measuring increased levels of known pro-apoptotic factors.

          To profile miRNA expression by hybridizing total CGN RNA with a custom microarray containing the complements to murine miRNAs.

Two anticipated findings were that miRNAs overexpressed in apoptotic neurons might be pro-apoptotic factors, while miRNAs with decreased expression might be anti-apoptotic. A number of additional preliminary experiments needed to be performed to optimize apoptotic conditions for the CGNs. Unfortunately, data from microarray experiments were not reproducible and therefore specific miRNA expression patterns could not be analyzed.

 

Effect of AMPK Activation on Insulin-Like Growth Factor-I Signaling in Vascular Smooth Muscle Cells

 

Adrienne Turner Duffield, David R. Clemmons

University of North Carolina, Chapel Hill, NC

 

The mitogenic effects of IGF-I mediated cell signaling in vascular smooth muscle cells have been implicated in the development of atherosclerotic plaques. In order to stimulate protein synthesis and cell proliferation IGF-I initiates activation of the mTOR/p70S6K pathway. The energy-sensing enzyme adenosine monophosphate activated protein kinase (AMPK) has antagonistic activity to mitogenic factors, specifically via the alternative regulation of this pathway. AMPK activation via the compound AICAR as been shown to have an antiproliferative effect in vascular smooth muscle cells. AMPK is also activated by the kinase LKB-1 upon treatment of the cell with the antidiabetic drug metformin. The aims of this study were to determine the effect of IGF-I on AMPK activation and the effect of AMPK activation on IGF-I mediated protein synthesis (specifically on mTOR/TSC2 pathway activation) in porcine aortic smooth muscle cells (pSMCs). It was anticipated that AMPK phosphorylation would have an antagonistic effect on IGF-1 signaling pathways. An additional objective was to determine the effect of AMPK activation in cells maintained in low vs. high glucose. The pSMC’s were maintained in either low or high glucose DMEM until confluent and were then treated with metformin or AICAR prior to IGF-I stimulation. The cells were lysed and the clarified lysates analyzed via SDS-PAGE followed by immunoblotting with the appropriate antibody (e.g. pAMPK, p70S6K). Pending statistical analysis, our results indicate that AMPK phosphorylation is inhibited upon stimulation with IGF-I, and that the inhibitory effect of AMPK phosphorylation on mTOR and P70S6K phosphorylation decreases with IGF-I treatment. The suppression of AMPK phosphorylation occurs despite activation of the enzyme with either AICAR or metformin. The implication of these results is that the effect of IGF-I on this signaling pathway in smooth muscle cells is predominant. The mechanism for this inhibitory effect has not been elucidated, but a major question for future investigation is whether forced overexpression of activated AMPK will down-regulate the ability of IGF-I to stimulate protein synthesis.

 
 

Effect of language task difficulty on cortical activation in normally developing pediatric subjects

1A Pendyal, 2MM Berl, 3EK Ritzl, 2,5L Rosenberger, 2DA Weber, & 4,5WD Gaillard

 

1University of North Carolina at Chapel Hill School of Medicine, Chapel Hill, NC, USA

2Department of Neuropsychology, Children’s National Medical Center, Washington, DC, USA

3Department of Neurology, The Johns Hopkins Hospital, Baltimore, MD, USA

4Department of Neurology, Children’s National Medical Center, Washington, DC, USA

5Epilepsy Research Branch, NINDS, NIH, Bethesda, MD, USA

 
Abstract
 

Background: fMRI is a non-invasive technique used to examine the neural correlates of language, working memory, and attention in normal controls. Objective: Using fMRI, to determine the changes in activation patterns that arise from an increase in language task difficulty within normally developing children. Previous studies involving adult subjects have suggested that an increase in language task difficulty may result in increased cortical activation in key Brodmann areas. Methods: We studied 19 normal, healthy, right-handed volunteers (12 male, 7 female, aged 7.25-12.58 years). We used whole-brain 3T fMRI (EPI BOLD), and employed a boxcar design that consisted of multiple runs (of varying difficulty) of an auditory category decision task. We measured in-scanner behavioral performance, including accuracy and reaction time. Data were analyzed using SPM2; individual t-contrast maps were generated, displaying differences in activation between medium and hard runs for each subject. Group “hard minus medium” results were then displayed as a single-sample t-test. Results: Average true positive accuracy among subjects was 83% for the medium task level and 72% for the hard task level; average reaction times for medium and hard task levels were 1085 and 1187 milliseconds, respectively. As expected, we observed a main effect of matched task level and age on accuracy and reaction time. The single-sample t-test (p=.005, uncorrected) revealed significant activation in the right anterior cingulate region (AntCing, Z=4.07), left inferior frontal gyrus (IFG, Z=3.57), and left superior temporal gyrus (STG, Z=3.23). Conclusions: In normally developing pediatric subjects, an increase in language task difficulty results in increased recruitment of cortical areas. Left-hemisphere activation of IFG and STG may reflect additional activation of traditional language areas, as well as the hard level’s additional working memory and language processing demands. Similarly, Right AntCing activation may reflect greater attentional or motivational demands in the hard version of the auditory category decision task.

 
 
 
 
Sensitivity of FDG PET in Malignant Lung Nodules Based on Non-
Attenuation Corrected Images, Attenuation Corrected Images and SUV

Amir H. Khandani1,5, Alexander W. Rich3, Bahjat F. Qaqish4,5, Leonard A. Parker2,5,

Marijana Ivanovic1, William H. McCartney1,5

1Sections of Nuclear Medicine, 2Thoracic Radiology, Department of Radiology, 3MD Program, UNC School of Medicine, 4Department of Biostatistics, UNC School of Public Health, 5Multidisciplinary

University of North Carolina, Chapel Hill, NC

 
 

AIMS: To evaluate the sensitivity of FDG PET in malignant lung nodules based on the visual interpretation of attenuation corrected images, non-attenuation corrected images, and based on SUV. METHODS: 62 lung nodules in 54 patients with preoperative FDG PET were included. Nodules with more intensity that normal lung or the mediastinum on non-attenuation corrected and attenuation corrected images, respectively, were considered positive for malignancy. In evaluation based on SUV, an SUV>2.5 was considered positive for malignancy. RESULTS: 61/62 (98%) nodules were correctly identified as malignant on non-attenuation corrected images. In comparison, only 36 (58%) were positive on attenuation corrected images, and only 38 were positive based on an SUV>2.5. The probability of missing a malignant nodule based on attenuation-corrected images or on SUV correlated negatively with nodule size. CONCLUSIONS: Non-attenuation corrected images are more sensitive than attenuation corrected images or SUV in detecting malignant lung nodules. Thus, non-attenuation corrected images should be used in future studies evaluating the sensitivity and specificity of FDG PET in diagnosing malignancy in lung nodules of indeterminate origin.    

 
 

Gene Expression Profile of Hematopoietic Stem Cells During Regeneration

Ali Chhotani and Tannishtha Reya1
Department of Pharmacology and Cancer Biology
Duke University, Durham, NC
 

Hematopoietic stem cells (HSC) are able to proliferate and regenerate rapidly in response to damage of the hematopoietic compartment. To better understand the regulatory mechanisms that govern this response at the molecular level, we treated mice with the chemotherapeutic drug cyclophosphamide (Cy) and the cytokine granulocyte colony-stimulating factor (GCSF). Treatment with Cy results in the loss of actively cycling progenitor cells in the bone marrow, followed by rapid proliferation and regeneration of HSCs. When Cy is combined with GCSF, a synergistic increase in proliferation is observed. Using the Cy/GCSF model, a microarray analysis was performed. The microarray analysis was a four-arm study in which HSCs derived from untreated mice were compared to those derived from mice treated with Cy alone, GSCF alone, or a combination of the two drugs. Gene expression profiles were created for each of the three experimental groups based on differential gene expression compared to the untreated group. Nearly 75% of all genes differentially expressed in this experiment are uniquely attributable to the combined Cy/GCSF treatment. Real-time PCR provided confirmation of several differentially expressed genes, including a novel gene in the HSC regenerative process. Gene ontologies of the differentially expressed genes vary widely but intracellular and membrane structural proteins, metabolic activity, and cellular physiological processes account for nearly 45% of the differentially expressed genes. Finally, this study suggests that the drugs induce distinct expression profiles when administered individually and in combination.

 
 

Early Experience with da Vinci Robot-Assisted Nissen Fundoplication in Pediatric Surgery

Andrew Dobberfuhl­­1, Daniel von Allmen2

1School of Medicine, 2Department of Pediatric Surgery

University of North Carolina, Chapel Hill, NC

 
 

Traditional non-robotic laparoscopic Nissen fundoplication (LNF) is a well described surgical solution to gastroesophageal reflux disease (GERD) in adults as well as children. The da Vinci Robotic Surgery System enhances LNF surgery with a three-dimensional view into the patient, wrist-like instruments, tremor filtration, and motion scaling. This study is a report of our early experience with this system to perform LNF in infants and small children. Goals were to investigate the intra and post-operative feasibility of da Vinci robot-assisted LNF in pediatric surgery and examine the difference in operating room costs for robotic versus standard LNF. From April 2005 through June 2006, fifteen children (5 months to 15 years) suffering from GERD unresponsive to non-operative therapy underwent LNF using the da Vinci Robotic Surgery System. All cases were reviewed retrospectively for demographic, intra and post-operative content. Total cost per robotic LNF was compiled and compared with non-robotic norms at our institution. Surgery time from first incision to time of dressing, averaged 167 min (98 to 250 min), and included robot docking time. Mean surgery operating time for fundoplication alone was 138 min (n=6) and with gastrostomy tube 187 min. (n=7). Following surgery, the mean time to discharge was 60 hours (n=14). There were no intra-operative complications or surgical mortalities. Operating room costs with the da Vinci robot averaged $1,609 more, and necessitated 40 min. less room time to complete than conventional LNF. These preliminary results of our series suggest that the da Vinci robot helps overcome the inherent challenges of performing LNF in children; benefiting the patient with decreased anesthetic exposure and the institution with shorter procedure times. Based on the intra-operative and short term follow-up data that was available at the time of this study, da Vinci robot-assisted LNF appears to be an effective alternative to conventional laparoscopy.

 
 

Sodium at 24 Hours of Life in Very Low Birth Weight Newborns: Does it Correlate with Dehydration?

 
Ashley Brandon
University of North Carolina, Chapel Hill, NC

Very low birth weight infants, or VLBW infants, have particular physiological challenges in regulating fluids and electrolytes, especially around the time of birth. Physicians often follow serum sodiums of VLBW infants to identify dehydration and manage fluids and electrolytes accordingly. However, currently there is little data to endorse this practice as a clinically useful technique. The purpose of this study is to explore the efficacy of this practice by providing quantitative data. A retrospective chart review was performed on all inborn neonates at UNC Hospitals in 2004 weighing less than 1500g at the time of birth. Demographic data as well as data concerning fluid and salt administration, weight change, and serum sodium levels in the first 24 hours of life was collected and analyzed. Of the infants included in the study (n=127), only 5 infants (3.9%) were found to have dehydration in the first 24 hours of life and of those subjects, only 3 of the 5 infants had abnormally high serum sodium levels. In contrast, 23 infants (18.1%) in the study suffered from hypernatremia, or a serum sodium measuring greater than 145 mEq/L. This result suggests that hypernatremia is not a marker of dehydration. There is, however, a very strong inverse linear relationship between percent weight change in the first 24-hours and serum sodium levels (p<5.99*10-7), as well as a clear correlation between percent weight change and the total amount of sodium administered during the first 24 hours of life (p<3.52*10-4). The take home message for physicians is that there is a complex interplay between fluid administration practices, sodium administration (either intentionally or non-intentionally administered), and serum sodium values that all need to be carefully balanced in order to avoid the pathological condition of dehydration.  

 
 

Predicting Patterns of Heterotopic Ossification in Burn Patients

A.N. Hardy1, K.P. Kolappa1, M. Kidd3, B. Cairns2, C.S. Hultman2, E.G. Loboa4, and J.A. van Aalst2

1UNC Chapel Hill School of Medicine; 2Department of Surgery, UNC Chapel Hill; 3Department of Surgery,

University of Oklahoma; 4Joint Department of Biomedical Engineering, NC State University and

UNC Chapel Hill

 

Heterotopic ossification (HO), the presence of ectopic bone in soft tissues, is the most common condition of excess bone formation in humans. HO affects 3-5% of burn patients. Previous work with osteoinduction has shown that tensile strain in the 10-12% range can result in ectopic bone formation. A statewide trauma and burn registry was queried for patients diagnosed with HO between 2001 and 2006. A total of 25 patients with radiographically-confirmed HO were identified. The demographics of these 25 were compared to that of all admissions (500/yr). The average age of patients with HO was 45 (18-77) compared to an average of 23 years for all admissions. Seventy-six percent of those with HO were male (compared to 73% of total admissions); 56% were African-American (compared to 26%). The average length of stay (LOS) was 114 days (compared to an average LOS of 13.2 days for all admissions); average TBSA was 40% (compared to 9%). Seventy-two percent of injuries were thermal, 16% combined thermal and chemical, and 12% electrical (compared to 3% electrical burns among all admissions). Seventy-two percent sustained inhalational injuries (compared to 15% of total admissions). The most common location for HO was the posteromedial elbow (68%) followed in descending order by the hip, shoulder, ankle, and mid-tibial shaft. In conclusion, burn patients who are older, African American, with greater TBSA burn, with inhalation injury, with an electrical injury, and increased hospital LOS are at increased risk for developing HO. The posteromedial elbow is the most common location for HO formation. Elbow biomechanics in the face of burn contracture produces a tensile strain in the 10-12% range, which may account for HO formation at the posteromedial elbow.

 
 

Commonalities and Gender in the Patient-Physician Interaction

Cherri Hobgood1,2, Morgan Lasater, Ashley Davis1, Mari-Wells Hedgpeth1

1Medical School Office of Educational Development, 2Department of Emergency Medicine

University of North Carolina, Chapel Hill, NC

 

Background: Empathy is an essential component of the patient physician interaction in order for the physician to build rapport with the patient. The concept of “commonality” is an aspect of empathy where similar world views/life experiences of the patient and physician helps them understand each other and effectively communicate. In this study a commonality is defined as an interaction between a patient and physician where either the patient or physician initiates a personal story, idea or thought in one of several categories. Then, depending on the topic, a response that completes this type of interaction may be to share something new about the subject, provide new information, or give an opinion.   

Aims: Determine if differences in occurrences of commonalities between patient and physician during the patient interview change as a function of the treating ED physician gender.

Methods: Data were collected in UNC-CH Emergency Department between the hours of 8 am and 2 am daily. Patient-physician interactions were observed and commonality occurrences were recorded. Then, surveys were administered to patient and physician participants.

Results: Mean number of commonalities for interactions between female physicians-female patients was the greatest at 0.87 with significance of 0.01 and the next highest was for male physician-male patients at 0.58 with significance of 0.46. There was so significant difference when looking only at patient gender or only at physician gender.

Discussion: There was a significant difference in the means for interactions between the genders of the physicians when looking only at interactions with female patients which had a significance of 0.01. When looking at physician genders for all male patient interactions the data was not significant. Male patient-male physician group had the second highest mean still supporting the hypothesis that gender concordant interactions result in greater number of initiated commonalities. Increase in data may later show significance in the male patient population.

 

Time Dependent Reduction of iNOS Protein and Downregulation of mRNA in Primary Cultured Hepatic Progenitor Cells

Babar Fiza*, Natasha Wright§, and David Gerber§

*University of North Carolina School of Medicine, Chapel Hill, NC

§Department of Transplant Surgery, University of North Carolina, Chapel Hill, NC

 

The unique ability of mammalian liver to regenerate after a damaging insult has prompted considerable interest into the research of the various cellular pathways responsible for hepatic regeneration. The protein, inducible nitric oxide synthase (iNOS) and its product Nitric Oxide (NO) are two factors whose roles have been studied extensively in mature hepatocytes. iNOS has been shown to have both pro and anti-apoptotic effects on mature hepatocytes. In contrast, there have not been any investigations into the role played by NO in Hepatic Progenitor Cells (HPC). Hepatic progenitor cells are a small sub-set of the cellular mass of the liver and are activated if mature hepatocytes lose their regenerative ability after a hepatic injury. The aim of the study was to observe iNOS in HPC and to evaluate the role of nitric oxide on HPC apoptosis and differentiation. The study also aimed to asses the role played by various cytokines on HPC growth and differentiation. To evaluate the expression of NO in HPCs, Western Blot and RT-PCR analysis were conducted on cells incubated with and without the presence of cytokines TNF-α, TGF-ß, and IL-1ß. The results of the study indicate time dependent reduction of iNOS protein and downregulation of mRNA in HPCs cultured in regular media. The study also demonstrates induction of iNOS in HPCs by IL-1ß. These findings illustrate a possible role played by iNOS in HPC differentiation.

 

Pediatric HIV Clinic Chart Audit: Determining

the Correlates to Mortality

Bradley C Fetzer1, Mina Hosseinipour2, Irving Hoffman3

1UNC School of Medicine, 2UNC Project, Malawi, 3Department of Infectious Disease

University of North Carolina, Chapel Hill, NC

 

Studies performed on HIV positive adults in Malawi, where over 15% of the population is estimated to be HIV positive, have been beneficial towards prolonging life, improving clinical care, and improving delivery of Anti-retroviral Therapy (ART). Few studies exist for HIV positive children in the hopes of achieving such aims. The goals of this study are to determine predictors of mortality in children on ART who attend the Pediatric HIV Clinic in Kamuzu Central Hospital in Lilongwe, Malawi. We conducted a retrospective chart review of all children who had started ART between x date and May 2006. Variables included basic demographics, clinical presentation, WHO HIV Staging, and treatment relating to HIV and their outcome as of June 2006. Descriptive statistics and bivariate analysis was performed to evaluate associations according to vital status. Cox proportional hazard modeling was performed to identify independent predictors of mortality. 258 children started ART. 51% were female, 7% were under 18 months, 26% were 18 months to 5 years, and 54% were >5 years of age. Most were WHO stage 3 or 4. (55% and 36%, respectively). 41 died, 216 were alive, 1 transferred . The mean survival time was 7.90+- 0.18 months. On multivariate analysis, factors associated with mortality included Ill at Presentation (HR 2.7 80% CI 1.51, 10.450, Weight Loss (HR 4.0, 80% CI 1.5 10.50), and age <18 months (HR 4.62 80% CI 2.3, 9.2). Most (83%) of the death occurred in the first 3 months of treatment. Early mortality among children starting ART is most associated with illness at time of presentation and being <18months. Evaluation of infants and children, prior to illness, will likely result in improved outcomes. Early diagnosis of infected infants using DNA testing or p24 antigen testing may allow early initiation of ART in infants at higher risk of death.

 
 
 
 

The Role of Cell-Associated VEGF Isoforms on the Transmigration of Choroidal Endothelial Cells Across the Retinal Pigment Epithelium

B. King, P. Geisen, J.R. McColm, L. Peterson, M.E. Hartnett

Department of Ophthalmology, School of Medicine

University of North Carolina, Chapel Hill, NC
 
 

Purpose: In age-related macular degeneration (AMD), choroidal neovascularization (CNV) is often preceded by the migration of choroidal endothelial cells (CECs) across the retinal pigment epithelium (RPE). The aim of this study was to investigate the role of VEGF isoforms in CEC transmigration when contact is made with RPE.


Methods: For hypoxia studies, ARPE were exposed to 24 hours of 1% O2. Controls remained in 21% O2. mRNA concentration was determined by real time RT-PCR and protein determined by Western blot. To determine the change in VEGF189 protein expression when CECs made contact with ARPE, ARPE were plated on the undersides of transwell inserts (contacting) or in the wells of a 6-well dish (noncontacting). CECs were plated in the inserts. For the transmigration study, ARPE were incubated in CellTracker Red and plated onto the underside of inserts. After 72 hours, primary human CECs incubated in CellTracker Green were plated into each insert. Then, after 24, 48, or 72 hours, inserts were transferred to a new well and trypsinized. Green cells taken from the well were counted on a hemocytometer to determine the number of transmigrated CECs.


Results: Exposure to 1% O2 conditions induced increased expression of VEGF mRNA for all three isoforms compared to 21% O2 (5.60, 8.68 and 10.01 fold increase for VEGF121, 165, 189, respectively). VEGF189 protein expression was higher in ARPE grown in contact with CECs than in ARPE grown in noncontacting coculture (0.479 vs 0.328 pixel density). Transmigration of CECs was increased in contacting culture vs. solo CEC culture (2091 cells/0.33 cm2 vs 270 cells/0.33 cm2 p = 0.016).


Conclusions: This study provides evidence that expression of VEGF189 is increased when ARPE are grown in hypoxia or make contact with CECs and may play an important role in the transmigration of CECs, a step involved in CNV.

 
 

Background: Patient behaviors such as smoking, unhealthy diet and low physical activity have been shown to directly contribute to chronic diseases like diabetes and heart disease. Stopping unhealthy behaviors reduces the incidence of both the diseases and their complications. The process of modifying an unhealthy behavior works best with both patient willingness and physician assistance.

Aims: The project sought to find relationships between chronic disease diagnosis and both patient willingness to change and physician intervention.

Methods: Patients were recruited at non-acute visits to six primary care practices across NC. Upon completing a waiting room computer survey, those patients identified as having at least one risky health behavior were enrolled. A follow up telephone interview gathered further information about the patient and their encounter with the doctor.

Results: The preliminary data (n=108) showed no significant relationship between a chronic disease diagnosis and patient being in the “active” stage of behavior change. Physicians, on the other hand, appeared to have their behavior influenced by patient chronic disease status. Multiple chronic diseases showed statistically significant relationships with physician intervention at p<.10 (a more rigorous standard will be applied when to the updated data used for the poster presentation). The consistent trend, observed for nearly all conditions, was for physicians to be less likely to counsel chronic disease patients about unhealthy behaviors but to be more likely to provide printed information and to refer to community resources.

Discussion: The initial data suggests that while physicians focus time and energy on unhealthy behaviors in their patients with chronic disease, the patients themselves may not be as aware of the link between behavior and disease. The increased rate of printed materials and referrals given to chronic disease patients suggests that physicians target aggressive interventions toward certain persons with chronic illness.

 
 
 

Association between Uterine Artery Resistance and Low Birth Weight

Catherine Varner1, Amy Herring1, John Thorp2

1 Department of Obstetrics and Gynecology; 2 Department of Biostatistics, School of Public Health

University of North Carolina School of Medicine, Chapel Hill, NC
 
 

Ultrasound is a key component in the evaluation of pregnancy. With improving technology, the ability to evaluate uterine artery blood flow parameters may predict adverse birth outcomes, including low birth weight. OBJECTIVE: This study investigated the relationship between high-resistance uterine artery waveforms in pregnancy and birth weight. METHODS: Uterine artery Doppler recordings were obtained between 15 and 20 weeks gestation and subsequently between 24 and 29 weeks in a cohort of women who attended prenatal care clinics in central North Carolina (n= 870). Doppler waveforms were evaluated by a blinded reviewer for the presence of an early diastolic notch, representing a failure of circulatory accommodation in pregnancy and a sign of high resistance blood flow. RESULTS: The mean birth weight of infants of women with any notching detected (any artery, any visit) was 150 g less than women who had no notching (p=0.001). The mean birth weight of infants of women with notching at both 15-20 weeks and 24-29 weeks was 160 g less than infants of women with notching present at 15-20 weeks, but resolved by 24-29 weeks (p=0.01). The largest discrepancy in birth weights occurred between women who had bilateral notching at 24-29 weeks as compared to women with no notching at either visit [3053.3 g vs. 3339 g; p=0.001]. CONCLUSIONS:  These findings indicate arterial notching is associated with low birth weight infants, and this association is most pronounced when notching is present at both time points in pregnancy. This information may be of value in increasing our understanding of the pathophysiology of growth restriction in pregnancy and may identify at risk pregnancies warranting closer surveillance. 

 
 
 

Development of Novel Biosensors to Determine the Balance between Activation of Different Rho GTPase isoforms in Malignant Breast Cancers

 

Christopher Welch, Louis Hodgson, and Klaus Hahn

Department of Pharmacology
University of North Carolina, Chapel Hill, NC
 
 

Rho GTPases are a major class of proteins critical to the control of cellular locomotion and have been implicated as markers for metastatic potential in a number of cancers. Specifically, RhoC overexpression has been closely correlated to prognostic outcomes of invasive breast cancers and other carcinomas. Recent evidence has implicated the balance of activity between RhoA and RhoC in controlling metastatic potential by either inhibiting motility (RhoA) or promoting motility (RhoC). These proteins have profound differences in function despite being greater than 80% homologous. Traditional biochemical methods have been unable to determine the difference in behavior produced by these small amino acid sequence differences. This is likely because the two proteins are subject to spatio-temporal regulation in intact cells. Thus, we are developing biosensors of RhoA and RhoC activation that can be imaged simultaneously in the same cell. We are exploring RhoA and RhoC biosensor designs based on FRET and on dyes that fluoresce at wavelengths different than those of the FRET sensors. For the FRET biosensors, the RhoA/C protein is linked to a yellow fluorescent protein (YFP) and cyan fluorescent protein (CFP) FRET pair, and to the Rho-binding domain of the Rho effector protein Rhotekin. Upon Rho activation, the Rhotekin Rho-binding domain binds to Rho, affecting FRET.   The dye-based RhoC biosensor construct consists of the Rhotekin Rho-binding domain site-specifically labeled with a cysteine-selective dye that changes fluorescence when the domain binds to activated, endogenous RhoC. We have explored the use of different Rho-binding domains, and have completed the RhoA and RhoC FRET sensors. Progress towards a dye-based RhoC sensor will be described. We are beginning to examine RhoA/C dynamics in the invasive breast cancer cell line MDA-MB-231 and in non-invasive fibroblasts.

 

Validating the Mini-Cog as a Screener for Mild Cognitive Impairment in the Primary Care Setting

James J. Lah1,2, Nelson Kyle Steenland1,3, Courtney Melton1

1Alzheimer’s Disease Research Center, 2Center for Neurodegenerative Disease, and  3Department of Environmental and Occupational Health

Emory University, Atlanta, GA
 

Mild Cognitive Impairment is a public health concern because studies of MCI subjects have shown significantly higher annual conversion rates to dementia than cognitively normal individuals. Therefore, identifying those with MCI allows for initiation of treatment in the earliest stages of cognitive decline, which has been shown to provide more benefit in delaying the progression of the dementia. There are several challenges faced in the Primary Care Setting that make screening for cognitive decline difficult, and research has shown poor MCI detection rates in these settings. The goal of this project is to validate the Mini-Cog combined with the Functional Assessment Questionnaire (FAQ) as an efficient means of screening for MCI in the primary care setting. Three-by-three tables comparing “gold standard” diagnosis (made by a neurologist) vs. predicted diagnosis by Mini-Cog and FAQ scores were evaluated for accuracy and kappa statistics. The preliminary analysis includes 158 subjects. The Mini-Cog and FAQ combination accurately predicted 126 of the 158 diagnoses made by the neurologists, for an accuracy rate of 80%.Weighted Kappa=0.78. The study is still underway and conclusions are pending.

 
 
 

Aging and Arterial Changes in the Cerebrovascular Tree

 

Eric Hintz, Elizabeth Bullitt, Mark Van Horn, and Donglin Zeng

CASILab, Department of Surgery

 University of North Carolina, Chapel Hill, NC 27599

 
 

Functional and histological changes in the cerebrovasculature associated with aging have been well documented. These changes include a decline in cerebral blood flow and both decreased density and ultrastructural degradation of capillaries. However, changes in the macrovascular tree, such as number and size of radiographically-discernible arteries, have not received equivalent attention. We address this deficit using novel software to analyze magnetic resonance angiogram (MRA) derived images in forty healthy volunteers. Subjects fit into one of two age brackets, 18-29 or 60+, each of which were equally divided by sex. Head-only MRA images were acquired in standardized fashion, yielding images containing 512 x 512 x 120 voxels acquired at 0.5 x 0.5 x 0.8 mm3. Automatic vessel extraction, performed by previously-created software, yielded text files containing coordinates and diameter for each vessel at a density of ten data points per voxel.   Our program calculated the total number of arteries in each brain and, for each vessel, determined average diameter. Based on diameter, arteries were categorized into one of 20 equisized bins ranging in size from 0.257 to 3.073 cm, with bin 1 containing the smallest vessels and bin 20 containing the largest. We found that, compared to the younger group, the brains of older subjects contained fewer arteries (265.65 v 341.75; p = 0.0016). Older subjects were more likely to have vessels in the large-diameter bins; statistically significant differences were noted in bins 4-7, 10, and a conglomerate containing bins 11-20. Furthermore, despite having fewer vessels overall, the older subjects had over twice as many arteries in bins 11-20 as the younger subjects (124 v 59 vessels). Younger subjects, on the other hand, were more likely to have vessels in bins 1 and 2. Together, this data suggests that normal aging is associated with a pruning of the vascular tree and a compensatory dilatation of the remaining vessels. This finding raises important questions about the role of macrovascular insufficiency in pathologic conditions more prevalent in older patients such as dementia and ischemic stroke.

 
 
Smo Expression in Basal-Like Breast Cancers
 

Hann-Hsiang Chao1 and Charles Perou2,3

 

1School of Medicine, University of North Carolina at Chapel Hill,  2Department of Genetics, University of North Carolina at Chapel Hill, 3Department of Pathology and Laboratory Medicine, University of North Carolina at Chapel Hill

 

Background and aims: Patients who develop basal-like breast cancer have been shown to have higher mortality rates than the other breast cancer subtypes, and tend to be largely unresponsive to most current forms of therapy. The basal-like breast cancers are known to be a type of basal cell carcinoma. Smoothened (Smo)is a proto-oncogene in the Hedgehog and Wnt signaling pathways, both of which play important roles in development, proliferation and differentiation. Overexpression of mutant Smoothened has been associated with the development of basal cell carcinomas. UNC patient data revealed certain mutations (V270I and D25G) in the Smo protein in patients with basal–like tumors. The aim of this study was to assess whether the V270I and D25G mutations were germline variants or somatic mutations and to determine the significance of the changes in the Smo gene.

 

Methods: The Smo full-length cDNA was cloned into the pBABE.puro expression vector. Site-directed mutagenesis was used to generate the Smo mutations of interest, V270I and D25G. The three Smo clones, along with the corresponding empty vector, were transfected into the SUM102 cell line. SUM102 was chosen because it is a basal epithelial cell line with low endogenous Smo expression. mRNA was isolated from the transfected cell lines, amplified and labeled, and microarray analysis was performed to create gene expression profiles. Significance Analysis of Microarrays (SAM) was carried out to determine significant gene expression changes caused by Smo overexpression.

 

Results: Smo expression was confirmed to be increased in all of the Smo transfected cell lines. A 2-class SAM showed that there was in a change in gene expression profiles when Smo was overexpressed versus the empty control. In addition, sequencing of a normal breast tissue sample from a patient revealed that the V270I change was present in normal as well as in the tumor sample.

 

Conclusions: Smo transfected SUM102 lines had increased Smo mRNA levels. Overexpression of Smo results in changes in the gene expression profile. The presence of the V270I mutation in a normal sample suggests that this change may be a germline variant.

 
 

Evaluation of Safety and Efficacy of an Insulin Infusion Algorithm

(Subproject under IRB 04-MED-358)
Hui-Jeong Song
School of Medicine
University of North Carolina, Chapel Hill, NC
 
Abbreviations:

MR=maintenance rate; IR=insulin infusion rate; ROD=rate of descent of blood glucose; MRcsne = MRcross step next estimate

 
Background:

Several clinical trials have shown that when patients with hyperglycemia were treated with insulin infusion, there was improvement of patient outcomes. The rigorous control of hyperglycemia reduced morbidity, mortality and hospitalization time for inpatients. The purpose of an insulin infusion protocol is to improve blood glucose control in hospitalized patients without the risk of substantial hypoglycemia. It would be an improvement if an insulin infusion algorithm could predict insulin requirements proactively rather than reactively for better glycemic control. It is known that both directional and rate changes of blood glucose influence the rate determination of insulin infusion. The purpose of the current study is to assess safety and efficacy of an Insulin Infusion algorithm of IRB 04-MED-358 protocol to estimate maintenance insulin requirements.

 
Methods:

Evaluable timepoints from 27 historical runs among trauma service patients were used to determine intervals of blood glucose stability based on the criteria set for the current study to help the consistency of the data analysis. Mean IRnext, historical during each interval of stability was determined. Values of MRcsne just before stable intervals were calculated according to the new algorithm:

 MRcsne= (IRprevious ) / {1 + [(RODprevious * 24hr) / (1800 mg/dL)]  

 
Results:

A total of 30 stable intervals were determined for the analysis. Data analysis revealed that there was a linear relationship between values of mean IRnext, historical during stable intervals and MRcsne just before the stable intervals. Strong correlation was observed between the values, R2=0.884, p<0.05.

 
Conclusion:

The study showed that hypothetical MRcsne values were systematically higher than mean IRnext, historical. Although the overestimation of mean IR on stable interval was made by MRcsne, the strong correlation between the two sets of values suggest efficacy and safety of the new programmable algorithm compared with the paper protocol used in the patient care.

 
 
Mapping the “Mighty Mini-muscle” Allele

John Hartmann1,Theodore Garland, Jr.2, Daniel Pomp1, and Gloria Munoz1

1University of North Carolina, Department of Nutrition, Chapel Hill, NC 27599, USA

2Department of Zoology, 430 Lincoln Drive, University of Wisconsin, Madison, WI 53706 -1381, USA

 
 

BACKGROUND: A collaborative project is currently being carried out to elucidate key mechanisms of locomotor adaptation and the physiological limits of selection. Selective breeding for high voluntary wheel running in mice has resulted in multiple lines that run an average of three times more per day than control lines. A key discovery of this selective breeding is the “mini-muscle” allele that has been favored and has gone to fixation in one of the high-running lines (S3). This autosomal recessive gene causes a 50% reduction in hindlimb muscle mass while doubling mass-specific aerobic capacity and altering contractile characteristics in ways that are hypothesized to enhance endurance running and the energetic efficiency of locomotion, but compromising sprinting abilities. My research goals consisted of identifying the chromosome harboring the putative mini-muscle locus, mapping it to a 3-5 cM region, and identifying possible candidate genes. METHODS: The mapping was performed through microsatellite marker analysis on a population of mice resulting from a backcross of S3 x C57 F1 to S3, resulting in a population with 50% affected mice. An initial genome-wide scan was done across selected backcross mice (N=54) to identify the chromosome harboring the mini-muscle gene. This was followed with fine mapping, which utilized the entire backcross population (N=404) to localize the gene to a manageable region. RESULTS: The genome-wide scan revealed that both of the two microsatellite markers run on CHR11 were significantly linked to the gene of interest (p<0.005). Fine mapping placed the gene in a final region of 2.634 Mb (67.455 Mb to 70.089 Mb) on CHR11. CONCLUSIONS: Further research may pinpoint the causal mutation, which may prove to be an important model for understanding basic muscle growth and development. Two candidate genes in this region were sequenced (SOX15 and CHRNB1), but the causal mutation has not been found. 

 
 

Proliferative subpopulations in partially differentiated embryonic stem cells: Characteristics and engraftment efficiency within the liver parenchyma.

 

Jonathan Haywood, Montserrat Caballero, Mai Nguyen, Suzanne Lyman and Jeff H. Fair

Department of Surgery – The University of North Carolina at Chapel Hill

Funding Provided by the NIDDK
 

Embryonic stem (ES) cells are pluripotent cells with capability to differentiate into cell types from the three germ layers, making them excellent candidates for cell replacement therapies. Although they represent probably one of the best therapeutic tools for many common diseases, there are still a lot of problems associated with their potential, such as lack of function, teratoma formation, and allograft rejection (1-4). Recently, in our lab, partially differentiated mouse ES cells have proved to engraft into the liver and reverse the hemophilia defect in a FIX knockout mouse model (5). Although the teratoma ratio in those experiments was low (6.2%), the risk is still present. The aim of this specific project was to characterize two subpopulations based on their expression of the tyrosine kinase growth factor receptor CD117 (c-kit), a proliferation marker present in some adult stem cells. Mouse ES cells were grown and differentiated in the presence of acidic fibroblast growth factor (FGF), and then sorted using magnetic beads on the basis of CD117 expression. The gene expression of positive and negative subpopulations was analyzed by qRT-PCR. Each population was then injected into the liver parenchyma of wild type mice to determine engraftment efficiency as well as teratoma formation rates. Gene expression analysis showed that endodermal markers are highly expressed in the CD117 positive fraction. The in vivo experiments showed that while CD117 positive cells induced teratoma formation in very high rates, the CD117 negative fraction did not engraft at all. The CD117 positive fraction is linked to a very high rate of teratoma formation, although the high levels endodermal markers would make them good candidates for robust engraftment. On the other hand the negative fraction does not engraft at all. Further subfractioning of the CD117 positive population is needed in order to select the population with high endodermal markers that would engraft without inducing teratoma formation.

 
 

Failure of Dexmedetomidine (DM) to Facilitate Extubation in a Trauma Intensive Care Unit (TICU)

Lauren Paton1, Craig Barrett1, Jonathan Salahshour1,2, Aram Kim1,2, Britt Christmas1, David G. Jacobs1, and Ronald Sing1

1Surgery, F.H. Sammy Ross Jr. Trauma Center, Carolinas Medical Center, Charlotte, NC, USA

2University of North Carolina at Chapel Hill School of Medicine, Chapel Hill, NC, USA

 
 

Introduction: Dexmedetomidine (DM), a selective alpha-2 adrenergic agonist that exhibits sedative, analgesic, anxiolytic, and sympatholytic effects to facilitate extubation. Present FDA recommendations allow DM administration for less than 24 hours with the dose less than 0.7 mcg/kg/hr. We retrospectively evaluated the use of DM to facilitate extubation for patients in the TICU.

 

Hypothesis: DM does not facilitate successful weaning and extubation from mechanical ventilation.

 

Methods: After IRB approval, TICU patients from January 2003 to June 2006 who received DM were retrospectively reviewed. Data collected included demographics, length of stay (LOS), days requiring mechanical ventilation, and number of hours receiving DM prior to weaning and extubation from mechanical ventilation.

 

Results: Sixty-six patients (55 males: 11 females) with a mean age of 41 years (range; 17-79) received DM while requiring mechanical ventilation. Average ICU LOS was 21.4 days (range; 1-54). Intravenous infusion of DM commenced at 0.4 to 0.7 mcg/kg/hr with a loading dose of 0.4 mcg. Background sedation and analgesia with propofol, benzodiazepines, and opiates were discontinued or reduced as tolerated. DM infusion was titrated between 0.2 and 1.9 with an average maximum dose of 0.83 mcg/kg/hr to maintain a stable cardiopulmonary response and modified Ramsay Sedation Score between 2 and 4. Only thirty patients (45.5%) were successfully extubated with the average time on DM till extubation being 58.9 hours requiring on average 0.91 mcg/kg/hr of DM. Only 4 patients (6%) were extubated within 24 hours of therapy and on less than 0.7 mcg/kg/hr. Average duration of therapy of DM was 74.9 hours (range; 1-302).

 

Conclusions: Under the current FDA recommendations for DM therapy, only 6% of patients would be successfully weaned. Challenging the current recommendations by extending the duration of therapy and increasing the dose significantly increases the number of successful extubations with this medication.

 
 

Whole Globe Enucleation Versus In Situ Corneal Excision:

A Retrospective Study of Tissue Trauma

Joseph Hoyle1, W. Craig Fowler2,3, and Mark Soper3

1School of Medicine 2Department of Ophthalmology, and 3North Carolina Eye Bank

University of North Carolina, Chapel Hill, NC
 
 

Purpose: To examine whether an enucleation (EN), followed by controlled laboratory excision, is a superior technique compared with a bedside in situ excision (IS) to minimize damage of corneal donor tissue before use in penetrating keratoplasty (PK) transplant surgery.

Background: Several small studies have suggested that the IS protocol is associated with increased tissue trauma and contamination. Nevertheless, during the last decade, IS has become the standard protocol because of lower costs to eye banks and a reduced time from death to placement in preservative media. 

Results: Records of all 1026 tissues procured by North Carolina Eye Bank (NCEB) personnel in 1998 were studied, and procurement protocols for 727 tissues were compared. Tissue exclusions were made because tissues were not used for PK (291), biomicroscopy data was not available (4), or the corneas were from infants (4). Analyses included endothelial cell density (ECD) and two retrospective tissue grades, assigned by the NCEB medical director, based on the cornea tissue evaluation forms originally completed by Certified Eye Bank Technicians (CEBT). One grade, the initial grade, was based on evaluation made shortly after excision of the cornea. The second grade, the final grade, was based on evaluation made usually twenty-hours later. We found significantly better ECD (3% of mean) and initial grade (41% of mean) for the EN protocol. There was no significant difference in final grade between the two protocols.

Conclusion: Our results support previous findings of increased tissue trauma associated with the IS protocol. The EN protocol may provide surgeons with improved tissue quality, although the tissue appears to recover.

 
 
 

Sodium-Calcium Exchange in HIV Neuropathogenesis

Karen Rusak, Rick B. Meeker

Department of Neurology, University of North Carolina, Chapel Hill, NC

 

Human Immunodeficiency Virus (HIV) rapidly penetrates into and infects the central nervous system (CNS). Although neurons are not infected, cognitive-motor deficits and neuronal loss are a significant consequence. The interaction of HIV with macrophages and microglia in the nervous system is thought to produce neurotoxic factors that are a primary cause of the neurological dysfunction. One of the proposed effects of the neurotoxic factors is dysfunction of the sodium-calcium exchanger (NCX). The NCX is responsible for the high capacity clearance of calcium from the cytosol using the sodium gradient to exchange sodium for calcium. Previous data suggest that decreased NCX function is a principle cause of the loss of calcium homeostasis following exposure to a variety of HIV-associated toxins, but the cause of the NCX dysfunction is unknown. We hypothesize that proteases such as metalloproteinases (MMPs) and calpain, released or activated during inflammation, may be responsible for the damage to NCX. NCX damage subsequently results in a loss of calcium homeostasis in neurons, neurological impairments and ultimately neuronal loss. To study this, an in vitro analysis was performed to examine calcium uptake into “inside-out” membrane vesicles prepared from rat or cat neural tissue and treated with medium obtained from monocyte derived macrophages (MDM) and metalloproteinases (MMPs). The radioisotope Calcium-45 was used to measure the amount of calcium influx into the membrane vesicles. Kinetics of NCX activity was examined, and MAP-2 staining of rat neuronal cultures was used to characterize the toxicity of MDM medium before and after treatment with the HIV envelope protein gp120. The results showed that neurons exposed to the MDM medium exhibit acute increases in intracellular calcium followed by more gradual long-term changes. Kinetic analysis to estimate the vesicular uptake of 45Ca revealed that treatment of the membrane vesicles with MMPs and MDM led to a decrease in Vmax. A significant decrease in Map-2 staining intensity was also seen after treatment of rat primary neuronal cultures with MDM medium, post treatment with gp120. All of these results suggest that the NCX was damaged by factors released from the MDM that was treated with gp120. This damage to NCX is likely a common cause of neuronal dysfunction in response to inflammation.

 
 
 
 
 

Synaptogenesis and Synaptic Elimination Across Development in Mouse Frontal Cortex

Karissa L. Gable, Chistina Grobin, Swarooparani Vadlamudi, John H. Gilmore, L. Fredrik Jarskog

 

Schizophrenia Research Center, Dept. of Psychiatry, University of North Carolina – Chapel Hill, Chapel Hill, NC

 
 
 

Background Schizophrenia is believed to be a neurodevelopmental disorder that involves both genetic and environmental causes. Aberrant synaptic change, specifically during synaptic pruning in adolescence, is a possible mechanistic theory for the development of schizophrenia.

Aims Characterization of cortical synaptic changes in a mouse model measuring both dendritic spine density and pre-and post-synaptic protein changes will give a thorough indication of normal development.   

Methods Mouse brain tissue that has been previously homogenized will be used in Western blot to assess protein levels of synaptophysin and spinophilin. Thy1 transgenic YFP mouse brain tissue will be coronally cut, mounted and imaged to assess dendritic spine density in the cingulate and hippocampal region using the Neurolucida imaging system.

Results Western blots showed evidence of a significant decrease in excitatory spinophilin across development, which then reached a plateau at 70 days. The synaptophysin protein, which is a marker for both excitatory and inhibitory synapses increased across development. Dendritic spine density is still in the process of measurement and analysis.

Discussion It was expected that synaptophysin would peak at adolescence and then decrease over development as has previously been shown in human tissue. The synaptophysin increase in mice could be explained in conjuction with the spinophilin results. In consideration of the fact that synaptophisin is a marker for all synapses, whereas spinophilin represents only excitatory synapses, the results of a decrease in excitatory synapses could represent a possible increase in inhibitory synapses. This would be consistent with recent work done with electron microscopy. The dendritic spine density analysis will be helpful in futher elucidating an explanation of the results.   

 
 
 

The Oxygen Dependent Regulation of ASB4 by FIH

Kevin R Smith1, J E Ferguson, III1,2, and Cam Patterson1,2,3,4

1Carolina Cardiovascular Biology Center, 2Department of Pharmacology, 3Department of Medicine and 4Department of Cell and Developmental Biology

University of North Carolina, Chapel Hill, NC

 
 

            Angiogenesis has importance in embryonic development and pathologic states in adult humans. The main regulator of angiogenesis is oxygen tension, which is indicated in the post-translational modification of the transcription factor, hypoxia-inducible factor-1α (HIF-1α). Under normoxic conditions, HIF-1α is hydroxylated at proline residues and an asparagine residue by prolyl hydroxylases and factor inhibiting HIF (FIH), respectively. This hydroxylation leads to a decrease in the stability and activity of HIF-1α, blocking the transcription of genes involved in glycolysis and angiogenesis, which are upregulated in response to hypoxia. Importantly, post-translational modification can occur at a faster rate than transcriptional regulation. Until recently, FIH was believed to only hydroxylate HIF-1α and its homologue HIF-2α. There is now new evidence that FIH has additional substrates for hydroxylation. Ankyrin repeat and SOCS-box protein-4 (ASB4) is a nine ankyrin repeat containing protein with a C terminus SOCS box that recruits the ubiquitin machinery leading to substrate protein degradation. ASB4 shows a narrow time frame of embryonic expression in cells of endothelial lineage, during which time the primitive vascular plexus is undergoing remodeling. We show that ASB4 binds to FIH through an EVN motif and mass spectroscopy is used to show that asparagine-246 in the EVN motif of ASB4 is hydroxylated by FIH in a manner similar to HIF-1α. The hydroxylation of ASB4 appears to show that hypoxia has a broader mechanism of controlling vascular development than simply transcriptional regulation. Therefore, it is likely that ASB4 functions to degrade a yet unknown substrate whose function is critical in the hypoxia regulated vascular development, and whose cellular levels require careful titration. Angiogenesis is important in disease states such as the formation of tumor vasculature, diabetic retinopathy, and ischemic heart disease. Therefore, we hope our results may lead to the identification of novel therapeutic targets for these diseases.

 
 

Regional Change in Brain Morphometry in Schizophrenia Associated with Short Term Antipsychotic Treatment.

Khary Carew1, Stacy Greeter1 and Robert McClure, MD2

1 School of Medicine, 2Department of Psychiatry

University of North Carolina, Chapel Hill, NC

 
 

Previous studies have shown volume changes in basal ganglia and cerebral cortex of schizophrenia patients undergoing long-term treatment with typical and atypical antipsychotic medications. The aim of this study was to identify if caudate and cortical volume changes occur over short periods of time (three months) in schizophrenia patients treated with typical and atypical antipsychotic medications. Patients satisfying DSM-IV criteria for schizophrenia were withdrawn from medication for three to six weeks. MRIs were performed after medication withdrawal and after twelve weeks of typical or atypical antipsychotic treatment. Region-of-interest (ROI) analysis was performed. Absolute volume changes measured by ROI were in the expected directions based on previous longitudinal studies. Caudate volume of typically-treated patients increased and caudate volume of atypically-treated patients decreased or did not change. The caudate results were not statistically significant, perhaps due to small sample size. Our results suggest that brief periods of treatment with antipsychotic medications are associated with changes in caudate volume.  

 
 

Role of NF-κB in osteoblastic differentiation, an implication of diabetic inflammatory disorder in bone healing

Lan Chi Tran1, Justin Barbaro2, Lyndon Cooper2,3, and Sompop Bencharit2,3,4

­1School of Medicine, 2School of Dentistry, 3Department of Prosthodontics, 4Department of Pharmacology

University of North Carolina – Chapel Hill

 

Type II diabetes patients have increased levels of inflammation and cellular apoptosis which can be induced by tumor necrosis factor (TNFα) signaling. The dysregulation of TNFα potentiates  inflammatory responses and induces apoptosis of matrix-producingcells. TNFα have been demonstrated to inhibit osteoblast differentiation via NF-κB pathway. However, NF-κB has also been shown to be implemented in the early process of osteogenesis. Interestingly, there is a concomitant upregulation of TNFα and IκBα, an inhibitor of NF-κB. The precise role of NF-κB and its inhibitor, IκBα, in the process of bone remodeling is still unclear. This study aims to elucidate the effects of NF-κB-IκBα interaction on osteogenic pathways. Human mesenchymal stem cells were transduced to overexpress IκBα or NF-κB, and then induced to differentiate into osteoblasts in vitro. For each line, the mRNA expression levels of several bone markers and cytokines were evaluated via SuperarrayTM Osteogenic analysis at timepoints 1, 3, and 7 days. The results from the gene array study will categorize osteogenic pathways that are upregulated by NF-κB and IκBα. Transduction of NF-κB upregulates most genes involved in osteoblastic differentiation, which include ICAM1, MMP10, type 2A1 collagen, TNFα and BMPs. There are little changes however in the expression of these genes in the control group and the one transduced with IκBα.. On the contrary transduction of hMSCs with IκBα downregulates several genes noticeably type 1A1 and 1A2 collagen and BMP6. These results imply that NF-κB may regulate cellular adhesion, matrix formation, and mineralization of osteoblastic stem cells.

 
 
 
 

       Lateral aggregation in BβA68T recombinant fibrinogen using a

    cell-based system

       Lance Warren1, Robert Campbell2, Susan Lord2, and Alisa Wolberg2

1M.D. Candidate, 2Department of Pathology and Laboratory Medicine,

                 University of North Carolina, Chapel Hill, NC, USA

 
 
 

Although very rare, patients with the BβA68T fibrinogen mutation (Fibrinogen Naples) display a high incidence of thrombotic episodes. Despite this phenotype, in-vitro clotting assays have shown that the final clot structure for Fibrinogen Naples is similar to wild-type when the same concentration of thrombin is used. The goal of this project was to utilize a model system assay to simulate the in-vivo coagulation process to compare clot formation, the end clot structure, and thrombin generation at onset of clot formation between wild-type and BβA68T fibrinogen. The model system is an assay that monitors clot formation in the presence of plasma concentrations of tissue-factor bearing monocytes, platelets, and blood clotting factors. The formation of clots was assayed within this model system for the mutant and WT fibrinogen utilizing a spectrophotometer that monitored changes in absorbance as a function of time. In addition, the formation of thrombin was assayed and used to determine the thrombin concentration at clot formation under each of the conditions. The end structure of the clots was then examined using scanning electron microscopy. The model system experiment was carried out three times over the course of the summer. For both WT and the BβA68T fibrinogen, three conditions were assayed: normal, elevated prothrombin, and hemophilic. In all three of these conditions and for each of the three model system experiments, the final turbidity of the BβA68T fibrinogen was lower than for wild-type. In addition, there was a trend of increased thrombin concentration at the onset of clot formation for the fibrinogen variant BβA68T. The lower turbidity for the BβA68T fibrinogen indicates thinner fibers and/or more tightly woven fibers which is consistent with the phenotype of increased thromboses in patients with this mutated fibrinogen. In addition, the onset of clot formation at a higher thrombin concentration may be the basis for the thinner fibers.

 
 
 

A Comparison of Fertility Correlates on the Guatemalan Agricultural Frontier

Laura P. Boschini

School of Medicine
University of North Carolina, Chapel Hill
 
 

This thesis conducted a comparison of fertility correlates in a population located on the Guatemalan agricultural frontier. The primary goal was to compare the effects of four groups of factors theorized to affect fertility: sociodemographic environment, openness to contraceptive use, degree of partner interaction, and couples’ desired fertility. Respondents used in this study were from communities on the agricultural frontier in and around the Sierra del Lacandón National Park in Petén, Guatemala. The analysis proceeded in two stages. A descriptive analysis using a series of means, medians, and ranges was first conducted to describe the study population. Five models using the different variable groups were then created and run using Poisson regressions. Model 1, the core model of traditionally-considered fertility determinants, contained only the sociodemographic variables. Each other variable group was then added individually to determine if that group improved the predictive value of the core model. Model 2 added the contraception variables. Model 3 added the partner interaction variables. Model 4 added the fertility desires variables. Model 5 contained all four groups of variables. The five models were then compared using a Bayesian information criterion. The population in question, a young, agricultural population of recently arrived migrants, possessed the high fertility and low prevalence of contraceptive use typical of an isolated frontier environment. A comparison of the five different regression models indicated that sociodemographic characteristics alone were most associated with fertility in this study population, followed by a combination of sociodemographic and partner interaction characteristics and by a combination of sociodemographic and fertility desires characteristics. Sociodemographic indicators seem to be most closely associated correlates of fertility in this population, although the strong showing of partner interaction and fertility desires variables may indicate close relationship that could not accurately measured in this study.

 
 

Predictive model for mediastinal lymph node status at the time of mediastinoscopy

Lauren Gainor1, Alden Parsons2, Leonard A. Parker3,
Frank C. Detterbeck2, D. Neil Hayes4

1UNC-Chapel Hill School of Medicine, 2Department of Cardiothoracic Surgery , 3Department of Radiology, and 4Lineberger Comprehensive Cancer Center

University of North Carolina, Chapel Hill, NC

 
 

Mediastinal lymph node (N2) positivity in non-small cell lung cancer (NSCLC) patients is suspected based on imaging such as CT or PET scan, with confirmation by mediastinoscopy. However, the most accurate clinical information in predicting N2 status is controversial. We reviewed 147 candidates for NSCLC resection (2000-2005) who had clinical database information available and had undergone mediastinoscopy. Using suspected clinical predictors of mediastinal metastasis available prior to mediastinoscopy, we constructed a predictive model of N2 status. The largest N2 node short-axis diameter on CT was by far the most influential factor in the model. Three other predictors for N2 node positivity were significant (p<0.05) in univariate analysis: indistinct tumor borders and mediastinal invasion on CT, and mediastinal PET scan positivity. However, all were less influential than N2 size on CT. Using logistic regression, these factors can be used to predict probability of positive N2 biopsy in an individual patient. The resulting diagnostic test had a ROC (receiver operator characteristic) area of 0.80 and optimal sensitivity-specificity pairing of 75% and 73%. 35% (51/147) of patients analyzed had at least one N2 node positive at mediastinoscopy. 39% (57/147) of patients had PET scan data available, and 82% (120/147) had CT data available. Of available data in early-stage NSCLC patients, mediastinal lymph node size on CT scan was more important than PET scan or other CT scan findings in predicting probability of positive mediastinoscopy. A predictive model is useful in more accurately determining need for invasive staging by mediastinoscopy.

 
 

The “Altitude Enigma” in Gastric Cancer in Central America

 

Lindsey Wilfley, Ricardo Dominguez, 2, Chris Martin, Paris Heidt,1 Douglas Morgan1.

1 Medicine, University of North Carolina, Chapel Hill, NC, United States and

2 Medicine, Hospital Regional del Occidente, Santa Rosa de Copan, Honduras .

 

Background. Gastric cancer is the second-leading cause of cancer mortality worldwide, with significant geographic variability. Areas of high incidence include Asia and Latin America. Further variability has been observed between areas of high and low elevation, potentially between mountainous and coastal regions. This has been observed in Latin America in the mountainous areas of the Pacific rim, extending from Mexico to Chile. This may relate to the interplay of host genotypes, H. pylori infection, dietary, and environmental factors.

Our aim was to investigate the variability of gastric cancer incidence based upon municipality elevation in western Honduras. The mountainous region of western Honduras is identified as a region of high incidence of gastric cancer. The standardized annual incidence is 30 per 105 (Males, 39; Females, 21; per 105, 2001).

 

Methods. The Western Regional Hospital (WRH) gastric cancer registry was used to identify incident cases from western Honduras for 1996–2005. Municipality of residence and origin were determined; the latter was used, as little mobility was noted in the population. Census data was obtained from the Honduras National Statistics Institute (INE, 2001). Standardized municipality-specific gastric cancer incidence rates were calculated. Elevations were obtained from Falling Rain Genomics, Inc. Linear regression was used to evaluate the relation between altitude and gastric cancer incidence.

 

Results. Municipality-specific gastric cancer incidence rates were calculated for 95 municipalities in western Honduras. Thirty-six principal municipalities were identified with rates ranging between 2.7 and 31.2 cases per annum per 100,000 population, combined for males and females. The mean elevation of the municipalities ranged 307 – 1210 meters. Five high incidence municipalities, all at higher elevations, were identified with a combined annual incidence of at least 20 per 105.

The regression analysis suggests a borderline positive linear relationship between municipality incidence and altitude (P = 0.065, F = 3.63). The correlation coefficient confirms altitude is not the dominant factor to explain intra-regional differences in gastric cancer incidence (R = 0.31, R2 = 0.096).

 

Conclusions. Gastric cancer incidence had a borderline correlation with elevation within the mountainous region of western Honduras. The observed “Altitude Enigma” in gastric cancer in Latin America is multifactorial, and more complex than elevation alone. It may be related to differential risk factors between coastal and mountain populations, including genetic and dietary factors. Further investigation in Latin America is warranted.

 
 
 

Early Formation of the Optic Cup in Sox2 Hypomorphic/Null Mice

Elizabeth R. Hoffman3, Scott R. Hutton1, 2, B. Matthew Fagan1, 2,
and Larysa H. Pevny1, 2

1Department of Genetics, 2University of North Carolina Neuroscience Center, 3MD/PhD Program

University of North Carolina, Chapel Hill, NC

 
 

Haploid insufficiency due to mutations in SOX2, a transcription factor affecting differentiation and maintenance of identity of neural progenitor cell populations, accounts for approximately ten percent of human micropthalmia (small eye) and anopthalmia (absent eye). In Sox2 hypomorphic/null mice, a reduction in SOX2 levels to less than forty percent of normal results in variable micropthalmia, hypocellularity of the retina, underdeveloped lens, and absence or hyperplasia of the optic nerve. It is not known if the decrease in SOX2 in hypomorphic/null embryos causes these defects through the regulation of neural cell differentiation alone, or if structural development of the optic cup in early embryos is also affected. Morphological and molecular marker analyses were conducted on the developing optic cup in Sox2 hypomorphic/null embryos at E10. No difference in overall optic cup structure was seen in Sox2 hypomorphic embryos. Consistent with SOX2’s role as a factor affecting neural cell differentiation but not regionalization, changes were seen in neurogenic markers β-TubulinIII and SIX3, but not in RALDH2, a marker of regionalization in the developing eye. The results suggest that the morphogenic formation of the optic cup is not affected by a decrease in SOX2 levels.

 
 

NSAID Use and its Effects on Barrett Esophagus and Esophageal Adenocarcinoma

D. Locke Glenn III1, Ryan D. Madanick1, Joseph A. Galanko1, Ginny Sharpless1, Nicholas J. Shaheen1

1. Medicine, University of North Carolina, Chapel Hill, NC, USA

 

Gastroesophageal reflux disease (GERD) affects the lives of many American adults, and it can lead to a pre-cancerous metaplastic change in esophageal epithelium known as Barrett’s esophagus (BE). Many therapies have been tested to treat BE, and one that has shown promise in the prevention of esophageal adenocarcinoma is non-steroidal anti-inflammatory drugs (NSAIDs).  It is unclear, however, whether NSAIDs play a role in preventing the progression of GERD to BE, or from BE to adenocarcinoma. We assessed the history of NSAID use among subjects with BE followed at UNC Hospitals. BE was defined as the endoscopic finding of cephalad displacement of the squamocolumnar junction of the esophagus, with histologic confirmation of specialized intestinalized metaplasia on biopsies from the tubular esophagus.   NSAID exposure data was collected from patients via a standardized questionnaire, which assessed multiple over-the-counter and prescription NSAIDs. Relative frequency of NSAID use was captured, as was the temporal period of use in the patients life. Subjects were categorized in one of four categories describing their NSAID use: “never users,” “current users,” “former users,” and “sporadic users.”    Other covariates which might act as confounders of the associate between the use of NSAIDs and the presence of BE were also collected, including age, race, gender, smoking, and alcohol use. Analysis was performed using Chi square and Fisher’s Exact test. Multiple logistic regression was performed with cases/control status as the outcome variable, and NSAID use as the predictor variable, with covariates as described above. P values of < 0.05 were considered significant. Three hundred and sixty patients participated in the study. The proportion of current NSAID users did not differ between cases and controls (43% vs. 44%).   Similarly, when grouping current and former users together, and comparing them to sporadic or never users, there was no significant difference between cases and controls (58% vs. 57%).   Despite the strong negative association between NSAID use and incidence of adenocarcinoma of the esophagus, our results suggest no relationship between NSAID use and Barrett’s esophagus.   Based on these results, one might surmise that NSAIDs may prevent the development of dysplasia in BE, but not the development of BE itself.

 
 

The Influence of Race and Sex on Patient Reported Pain Levels

Mary Rogers and Cherri Hobgood1

1Curriculum and Educational Development

University of North Carolina School of Medicine, Chapel Hill, NC

 
 

Pain is often reported to doctors by patients on a 0 to 10 scale, with 0 being no pain and 10 being the worst pain imaginable. This scale is not precise because patients view pain differently from doctors. This could lead to physicians under or over treating patients. Physical characteristics about doctors and patients could influence the disparity.  This study focused on determining if the sex of physicians and patients or the race of the patient affected patient reported pain. Patients and physicians were recruited in the UNC-CH emergency department and were observed by a trained research assistant. After the doctor finished interviewing the patient, both research subjects received surveys that asked the patient’s pain level on the 0 to 10 scale along with numerous other questions. In total, 79 patients were interviewed. Analysis of the data indicated that there was no statistical significance showing that patient and physician sex or patient race changed how a patient reported their pain level. More research needs to be conducted to test and see if other characteristics like age affect how a patient reports their pain level. Better understanding of patient pain levels would help the medical field to give more appropriate treatment.

 

Protein C Pathway Abnormalities as Biomarkers of Hypercoagulability in SLE and APS

Lynn Howie,1 Jon Solow,1 and Robert A. S. Roubey1

1Thurston Arthritis Research Center, University of North Carolina at Chapel Hill

 
 

The Antiphospholipid Syndrome (APS) is the association of autoantibodies with thrombosis, pregnancy loss, and other clinical manifestations. APS is a common cause of acquired thrombophilia in the general population. In patients with systemic lupus erythematosus (SLE), thrombosis due to APS is an important cause of morbidity and mortality. Antiphospholipid antibodies (aPL) are likely contribute to hypercoagulability through multiple mechanisms. Predicting which patients with aPL are at greatest risk for thrombosis is important clinically. Presently it is not possible to identify who would benefit from prophylactic anticoagulation and spare those at less risk. We hypothesize that thrombosis in patients with APS is multifactorial, including abnormalities of the protein C pathway. 300 patients from the Antiphospholipid Syndrome Collaborative registry not on anticoagulant therapy at study enrollment were studied to determine if deficiencies in Protein C and/or Protein S were present. Frozen plasma was used for ELISA testing of Protein C, Total Protein S and Free Protein S (Diagnostica Stago). 6.3% of subjects had Protein C deficiency, 4.5% had Total Protein S deficiency and 21.3% had Free Protein S deficiency.  Total Protein S deficiency was associated with a history of arterial thrombotic events (OR 9.1, 1.7- 46.8), pregnancy morbidity and mortality (OR 5.8, 1.2 – 28.0), and classification as definite APS (OR 5.4, 1.1 – 25.8).  Protein C and Free Protein S deficiencies were not associated with a history of clinical manifestations. Deficiencies of protein C, total protein S, and free protein S are more prevalent in patients with aPL than in the general population. Deficiency of total protein S is associated with arterial thrombosis, pregnancy morbidity and mortality, and with classification of definite APS.  This cohort continues to be followed. Future studies will examine whether baseline visit abnormalities of the protein C pathway are associated with increased risk of future clinical manifestations.

 

Purines in Exhaled Breath Condensate as a biomarker of Inflammation.

H. Matias Jasin1,Leonard Collins2,Gunnar Boysen2 ,Charles R. Esther, Jr.3

1MS2, 2Biomarker Mass Spectrometry Facility, and

3Pediatric Pulmonology, University of North Carolina at Chapel Hill,

Chapel Hill, NC, USA.

 
 
 
 

Background: Assessment of inflammation in the lung has been a challenging problem in patients with cystic fibrosis (CF). Historically, lung inflammation has been measured by analysis of bronchoalveolar lavage fluid (BALF) attained through invasive bronchoscopies. Exhaled breath condensate (EBC) is a relatively new, non-invasive technique for collecting dilute samples of airway surface liquid (ASL). A highly positive correlation between neutrophil counts and purine levels in BAL samples has recently been shown. This indicates that purines may serve as good markers for neutrophilic lung inflammation.

Aims: Develop a method for detecting urea (a dilution marker), adenosine, AMP, and ATP in EBC samples using liquid chromatography/ tandem mass spectrometry (LC/MS/MS) and assess the differences between EBC purine levels in CF vs. non-CF controls.

Methods: Optimization of the electrospray LC/MS/MS method was performed using purine standards injected directly into the MS/MS. Trials were run to find the optimal buffer pH for ionizing and detecting purines. Standard curves at optimal pH were produced for estimating EBC purine concentrations. With a reliable method in place, EBC samples were collected from pediatric CF patients (n=9), asthma patients (n=3), and healthy controls (n=4). Levels of urea, adenosine, and AMP were measured using the optimized LC/MS/MS method.

Results: We developed a method to reliably detect urea, AMP, and adenosine. Our work showed that purines are best ionized in buffer systems near pH 8.5. For ATP, however, these results did not translate to better detection in samples run with the LC column in place. EBC results showed increased AMP/Ado ratios in the CF airway compared to control, though the result did not reach significance (p=0.057).

Conclusion: This work is a first step in showing that AMP can be reliably detected in EBC To further support these preliminary findings, larger cohorts need to be studied and further optimization of the method needs to be done to refine detection techniques for ATP.

 
 
 
 
 
 

Prostate cancer in men treated with testosterone replacement therapy

R. Matthew Coward, Jay Simhan, and Culley C. Carson III, MD

Department of Surgery, Division of Urologic Surgery

University of North Carolina School of Medicine, Chapel Hill, NC

 
 

Introduction and Objectives: Although the long-term effect of testosterone replacement therapy (TRT) is unclear, a theoretical increased risk of prostate cancer (CaP) is a primary concern in treating hypogonadism. We retrospectively reviewed hypogonadal men on TRT to evaluate the occurrence and disease progression of CaP, and further to analyze the dynamics of prostate-specific antigen (PSA), in this population. Methods: A total of 81 hypogonadal men, age 56.8 ± 11.5 (range 25-82), were followed for 33.8 ± 29.7 months after initiating TRT. Testosterone and PSA levels were assessed every 6-12 months. Results: Baseline PSA was 1.32 ± 0.98 ng/ml. Pre- and post-treatment (36 months) total testosterone was 241.1 ± 103.5 ng/dl and 379.8 ± 203.3 ng/dl (p=0.003), and free testosterone was 7.9 ± 5.3 ng/dl and 13.3 ± 7.2 ng/dl (p=0.015), respectively. Four men, age 54.5 ± 8.7, developed CaP an average 32.5 ± 7.9 (range 22-41) months after initiating TRT. Three men were diagnosed with Gleason 3+3 CaP and one with Gleason 3+4 CaP. Two men underwent radical prostatectomy with negative surgical margins and subsequent PSA nadirs <0.1 ng/ml. One patient elected external beam radiation therapy with a PSA nadir of 0.7 ng/ml. The fourth patient had not pursued therapy at the time of the study. There was an increase in PSA from baseline to 18 months of 1.8 (p=0.15), while the PSA from baseline increased by 3.2 (p=0.047) at 36 months. In men without CaP, the PSA did not increase significantly for 60 months. Conclusions: Although four hypogonadal men on TRT developed CaP, these cases were of moderate grade and were successfully treated. We recommend closely monitoring all men during the first three years and to consider a decrease in the threshold for prostate biopsy in those men who experience an increase in PSA from baseline over the same time period.

 
 

The Effects of the novel protein BARC on Breast Cancer Cells

Matthew Triplette1

1Department of Cancer Biology, Wake Forest University Health Sciences

 

BMP Antagonist Regulated in Cancer(BARC), may influence growth and progression of cancer cells in breast cancers as a BMP antagonist. Bone Morphogenetic Proteins (BMPs) can contribute to cancer progression through the TGF-beta pathway, with increased BMP levels leading to increased, and potentially unregulated, cellular growth. The objective of this study was to demonstrate that BARC has a significant effect on secreted levels of BMP-7 in breast cancer cell models, and determine if increased BARC, leading to downregulation of BMP-7, has a significant suppressive effect on breast cancer cell growth. The first objective was accomplished through a P-Smad immunoblot, detecting the amount of R-Smad phosphorylated extracellularly by BMP-7 in the TGF-beta pathway.   Phosphorylated-Smad levels were compared in BARC and control groups after adding a standardized amount of BMP-7 and the particular treatment to ductal carcinoma (MCF7) cells. To accomplish the second objective, BARC mRNA levels from cDNA samples were used to compare BARC in normal and tumor tissue, BARC cellular and extracellular protein levels were compared across the Weinberg breast cancer cell progression line using Western blot, and, utilizing tissue microarrays, normal and cancerous tissues from patients were probed with BARC and their images were compared by pixel intensity. The P-Smad immunoblots of MCF7 cells demonstrated that BARC downregulates BMP-7 to a further degree in these cells than known antagonist Noggin, the BARC mRNA levels demonstrate a significant decrease in BARC in tumor versus normal tissues in 81.1% of the patients, and the immunoblots of the Weinberg line demonstrate that the normal and low oncogene cells secrete significantly more BARC than cancerous cells. This preliminary study on BARC’s effect on breast cancer cells illustrates that BARC does act as a significant BMP-7 antagonist in the extracellular environment of breast cells, and its level of secretion has a significant effect on breast cancer cell growth.

 
 

Commensal microflora promote post-surgical IBD and fibrosis: Meagan Hunt, Rachael Rigby, Brooks Scull, Micheal Helmrath and P. Kay Lund.

 

Introduction: Crohn’s disease (CD) is an incurable, chronic inflammatory bowel disease (IBD). Fifty-80% of CD patients require bowel resection. Recurrent inflammation and fibrosis lead to repeat surgery in 20-65% of patients. Ileo-cecal resection (ICR) predicts the highest risk. Commensal microflora appear to initiate and perpetuate IBD in humans and the IL-10 null model of IBD. We developed a model of ICR in IL-10 null mice to test the hypothesis that commensal microflora promote post-surgical IBD. 

Methods: IL-10 null mice received ICR, or control (sham surgery or no surgery) while germ free (GF) and disease free, or 3 weeks after bacterial colonization (CONV), when inflammation is established in cecum. Animals were killed 28 days post-op. Small intestine (SI), neoterminal ileum/anastomosis (NTI), and colon, were scored histologically for inflammation and fibrosis. Post-surgical disease in SI and NTI was defined as scores higher than in mean CONV or GF controls.   Results: In 10/11 CONV-IL-10 null mice, ICR led to inflammation and fibrosis in SI (a site typically disease free) and NTI. ICR in GF-IL-10 null mice led to inflammation and fibrosis in 5/9 SI and 8/9 NTI, although mean disease scores were lower than in CONV-IL-10 null given ICR. Negative bacterial cultures, and inflammation scores below 1 in colon and SI of GF-IL-10 sham or no surgery controls, while co-housed with GF-ICR mice, confirmed GF status after surgery.

Conclusions : ICR in IL-10 null mice provides a new animal model of post-surgical IBD in small intestine and neoterminal ileum, sites of post-surgical disease in human CD. Differences between GF and CONV after ICR demonstrate that commensal microflora promote more severe and frequent post-surgical disease. Inflammation and fibrosis in a significant number of GF-IL-10 null mice after ICR indicate that bacteria-independent factors also promote post-surgical IBD. Our model should prove valuable for future analyses of mechanisms and preventive therapies for post-surgical IBD.

 
 
                                               
 
 

Regulation of the Myocyte Enhancer Factor 2A (MEF2A) gene by microRNA-346

Nikhil Jariwala, Mariko Tatsuguchi and Da-Zhi Wang

Department of Cell and Developmental Biology

Carolina Cardiovascular Biology Center

University of North Carolina, Chapel Hill, NC
 
 

The MEF2A gene is critical for muscle cell differentiation. Although it is known that regulation of the MEF2A mRNA depends upon the 3’UTR, the exact mechanism is not known. Current research suggests that miRNAs may cause this regulation due to their propensity to down-regulate mRNA translation by binding to the 3’UTR. Theoretically, miR-346 should regulate MEF2A by reducing gene expression in a reporter assay. While the strong seeding region of miR-346 suggests one particular binding site on the 3’UTR, the data is unclear as to whether miR-346 actually binds to this site. Future analysis with a Western Blot may help determine how miR-346 regulates gene expression.

 
 
 
 
 

Bayesian Clustering and GO Identity in Bone and Prostate Cancer Microarray Data

Omar Halawa1,2,3, Debashis Ghosh 2,3, Zhaohui Qin 2,3

 

1 University of North Carolina School of Medicine, Chapel Hill, North Carolina 27514. USA.

2 Bioinformatics Program, University of Michigan Medical School, Ann Arbor, Michigan 48109, USA.

3 Department of Biostatistics, University of Michigan Medical School, Ann Arbor, Michigan 48109, USA.

 

Microarray technology has spurted the growth of a new kind of science; a faster, higher throughput kind. Thousands of genes across hundreds of samples can be analyzed instantly. However, such large datasets and complex diseases such as cancer require a unifying technology. The field of gene clustering has proven to be the most successful in ascertaining biological relevant information from thousands of rows of gene expression. Here we apply a Bayesian gene expression clustering program developed by Dr. Qin named Chinese Restaurant Cluster (CRC) to analyze a hierarchy of microarray datasets. First, the method is applied to a bone cancer dataset, and then applied to a more complex in-vitro/in-vivo model of prostate cancer. We uncover that the Bayesian approach allows for a specific look at gene clusters, whereas the frequentist approach displays a “bird-eye view”. Moreover, we show the pertinent information ascertained from incorporating gene ontology into the Bayesian model. The future holds a place for a Bayesian clustering program that unifies a model based approach with systems biology, creating a more dynamic tool for understanding the genetic component of many types of cancer. It is by this approach that we hope to more efficiently uncover clinically relevant clusters of genes associated with many different types cancer.

 

The Relationship Between Functional Health Literacy and Adherence to Emergency Room Discharge Instructions Among Spanish-Speaking Patients

Patrick C. Smith, Jane Brice, MD MPH, James Lee, MD
Department of Emergency Medicine

University of North Carolina at Chapel Hill

 

As the Hispanic population in the state explodes, so does the number of Spanish-Speaking patients seeking medical care at the hospital emergency department (ED). Recent studies have shown that there is a disparity in health between immigrant populations and the rest of society. Other studies indicate that Spanish-Speakers have a lower rate of compliance with ED discharge instructions, thus adversely affecting their eventual health outcomes. One reason for lower adherence may involve the concept of Functional Health Literacy (FHL), which is defined as “the degree to which individuals have the capacity to obtain, process, and understand basic health information and services needed to make appropriate health decisions.” Our objective is to 1) further explore whether or not, compared with English-speakers, Spanish-speakers who present to the ED have lower rates of adherence to discharge instructions, measured by the rate of going to follow-up appointments and filling prescriptions, and 2) investigating if FHL is more closely associated with adherence in Spanish-speakers than in English-speakers. Fifty adult Spanish-Speaking Patients were interviewed while in the ED and their FHL was assessed using the Test of Functional Health Literacy of Adults in Spanish (TOHFLA-S). A gender and age matched English-Speaker was then interviewed using the TOHFLA in English. A second interview was then conducted by telephone to assess the compliance with medical instructions received upon discharge from the ED. All conversations and written material were done in each patient's first language by a bilingual investigator. Initial results confirm that Spanish-Speakers have a lower rate of compliance with ED discharge instructions. Further, those patients with a lower FHL had a lower rate of compliance than those with a higher FHL. 

 

CYCLOSPORINE DOES NOT PENETRATE, BUT ACCUMULATES AT HIGH CONCENTRATIONS WITHIN, DIFFERENTIATED HUMAN AIRWAY EPITHELIAL CELLS IN VITRO: POSSIBLE IMPLICATIONS FOR THE USE OF INHALED CYCLOSPORINE

Robert Aris, MD1, Patrick McNeillie, BS1, Olusegan Olusesi, BS1, Sarah Thomas, BS1, Mary Paine, MD2, Catherine Hammett-Stabler, MD3 and Isabel Neuringer, MD1.

1Pulmonary and Critical Care Medicine, University of North Carolina at Chapel Hill, Chapel Hill, NC, United States, 27599; 2School of Pharmacy, University of North Carolina at Chapel Hill, Chapel Hill, NC, United States, 27599 and 3Pathology and Laboratory Medicine, University of North Carolina at Chapel Hill, Chapel Hill, NC, United States, 27599.

 
 
 
 

Purpose: Recently, inhaled cyclosporine has been shown to reduce mortality and BOS. We hypothesized that inhaled cyclosporine is absorbed through the respiratory epithelium to exert a local immunosuppressive effect. We studied the kinetics of this process using a tissue culture model.


Methods and Materials: Human tracheobronchial epithelial cells (hTBE) from healthy donors were grown to confluence at an air-liquid interface. Differentiated hTBEs were treated on their apical surface with cyclosporine concentrations of 100, 1000, 10,000, and 100,000 ng/ml to mimic the effects of inhaled cycolsporine. The basilar, apical and intracellular compartments were assayed for cyclosporine by LC-MSMS.


Results: At apical concentrations of 100 and 1000ng /ml, cyclosporine was not detected in the basal compartment. At the apical concentration of 10,000 ng/ml, the median (95%CI) basal concentration was 0, 0, 85 (42-128) and 152 (132-172) ng/ml at 3, 6, 10 and 24h, respectively. At the apical concentration of 100,000 ng/ml, considerably more drug reached the basilar compartment but light microscopy revealed severe tissue injury and loss of barrier architecture. From 24 to 120 hrs, basal concentrations plateaued. Apical concentrations of cyclosporine fell 5 fold over 24 hrs and persisted at these levels out to 120 hr. Accounting for dilution, only 5-10% of the apically-deposited cyclosporine reached the basal compartment at the concentration of 10,000 ng/ml. Intracellular concentrations of cyclosporine were >100 fold above starting concentrations from 6 to 120 hr. These kinetics suggest a 2 compartment model. Western blots revealed that human airway epithelium did not express the CYP3A4 isoenzyme that metabolizes cyclosporine.


Conclusions: Cyclosporin is very inefficiently absorbed by airway epithelial cells and, interestingly, accumulates within these cells at high concentrations in vitro. Inhaled cyclosporine may work, in part, by altering epithelial cell function.

 
 

Fractionation, Dose selection, and Response of Hepatic Metastases of Neuroendocrine TumorsAfter 90Y - Microsphere Brachytherapy

Patrick McNeillie, BS1, Andrew S. Kennedy, MD, FACRO1, William A. Dezarn, PhD1, Carroll Overton, MD2, Mary England, RN1, and Scott L. Sailer, MD1

1Radiation Oncology, Wake Radiology Oncology, Cary, NC, United States, 27511; 2Interventional Radiology, Wake Radiology , Raleigh, NC, United States, 27511

 

This is a retrospective review of patients from a single institution that received 90Y- microsphere therapy for neuroendocrine hepatic metastases. Physical, radiographic, biochemical, and clinical factors associated with treatment and response were examined, including serial Chromogranin A (CgA) and liver function tests. All patients were followed with laboratory and imaging studies at regular intervals until death, or censured if other therapy was given after Brachytherapy. Toxicities (acute and late) were recorded, and survival of the group determined. We present novel approaches in the use of 90Y - microspheres for unresectable liver metastases from a variety of neuroendocrine tumors. Standard use of microspheres is to apply once, typically to a single lobe, followed a month or so later with treatment of the other lobe. We have used them differently, implanting all the tumors during each treatment i.e. whole liver. A second and third treatment was given to gain control of large hepatic lesions.

 

Eighteen consecutive patients (10M, 8F) received 24 fractions.The median follow-up is 27 months (4-44 mo). All had receivedprior therapy, with 16/18 chemotherapy, 6 TACE, 2 RGA, and 2lobar resection. Carcinoid symptoms were present in 13 patients prior to treatment, with objective responses found in 16 (89%) by imaging and CgA. Median survival has not yet been reached with only 2 patients succumbing to metastatic disease. There were no treatment related deaths; radiation induced liver ease, or veno-occlusive disease. Tumors included glucogonoma 1, atypical pulmonary 1, islet cell 1, insulinoma 2, and carcinoid from a small bowel primary 12, usually terminal ileum.

 

Whole liver and multiple fraction microsphere brachytherapy are safe,feasible, and produce a high response rate, even with extensive tumorreplacement of the liver. Acute and delayed toxicity was low without atreatment related Grade 4 acute event, or radiation induced liver disease in this small cohort. Nearly all patients experienced a significant objective response, suggesting that further investigation of this approach iswarranted.

 

Complete Radiographic Response of Hepatic Metastasesfrom Small Cell Lung Cancer

Treated with 90Y-Microspheres

Patrick McNeillie, BS1; Andrew Kennedy, MD2; Mary England, BS2; William A. Dezarn, PhD2

1Pulmonary and Critical Care Medicine, University of North Carolina at Chapel Hill, Chapel Hill, NC, United States, 27599; 2Wake Radiology and Oncology Service, Cary, NC United States, 27511.

 
 
Introduction:

Microsphere brachytherapy has become an effective treatment for primary and metastatic cancer to the liver. It is a way of permanently implanting liver tumors with radiation utilizing the arterial blood supply. The selective nature of this approach is based on the anatomic fact that 80-100% of the blood supply to

metastatic tumors in the liver derived their arterial supply from the hepatic artery, while the normal liver is predominately feed by the portal venous system. The functional unit is a microsphere (diameter 25-35 microns) containing the therapeutic isotope Yttrium-90 (90Y) contained within a matrix of glass or resin. We report a single, but not uncommon, case of complete resolution of hepatic tumors from a single application of microspheres.

                                       
Microsphere Therapy

While the rate of cancer in Americans has steadily declined since 1999 the rate among individuals age 65 and older has remained fixed at 24.8 per 100,000. This statistic is worrisome as the relative age of Americans increases due to the mid-1900 baby boom, and predicts for an increase in the number of

patients in need of effective therapy for hepatic metastases from all solid tumors. The current literature is limited in analyzing treatments of primary lung cancer with metastases to the liver. Nagashima published

a successful surgical series of liver tumors from lung cancer origin with some 5-year survivors. While surgical treatment may provide the best treatment for a very small select group of patients, Y-90 microspheres can provide a treatment option to a greater percentage of patients. For the current patient, it is possible that other local therapies could have been attempted, but microspheres have provided a significant benefit without surgical intervention, and without side effects.

 
Discussion

Hepatic metastases from a bronchogenic carcinoma can be controlled with a single, well tolerated brachytherapy procedure with Y-90 microspheres. For a percentage of patients with liverpredominant

metastases, microspheres may be a helpful adjunct to systemic chemotherapy in controlling potential deadly distant disease. Our patient, like the majority of microsphere patients, had little or no toxicity (subjective or objective) while deriving a radiographic complete response in the liver.

 

Visual Function after Minimally Invasive Pituitary Surgery (MIPS)

Kuruvilla R, Ewend MG1, Senior BA2, Givre SJ3,4

1Department of Neurosurgery, 2 Otolaryngology, Head and Neck Surgery, 3Ophthalmology, and 4Neurology

University of North Carolina, Chapel Hill, NC
 
 

Introduction: Preservation and restoration of vision are important goals for surgical removal of pituitary adenomas. As compared with traditional, sublabial transsphenoidal (SLTS) pituitary surgery, endoscopic endonasal surgery (MIPS) is reported to have fewer complications and result in shorter hospital stays. However, in order for MIPS to be accepted along side traditional approaches, visual outcomes following MIPS must be comparable to those from traditional surgery. Methods: Retrospective review of patients undergoing MIPS who had ophthalmologic examination and, when possible, visual field testing (automated threshold or Goldman perimetry) before and after surgery. Surgery was performed by a single team of surgeons. Pre- and postoperative data were compared. Automated threshold perimetry was scored using a system similar to that used in the Collaborative Initial Glaucoma Treatment Study. Results: The duration between surgery and post-operative ophthalmologic examination ranged from 2 days to 35 months (mean 6.1 months). Of the 35 eyes evaluated, initial post-operative visual acuity was unchanged in 17 (48%), better in 14 (40%), and worse in 4 (11%). Initial post-operative visual field scores were unchanged in 7 (20%), better in 24 (69%), and worse in 4 (11%). Only two eyes showed a decrease in both visual acuity and field. Averaged across all eyes, the postoperative visual field score after MIPS was significantly improved as compared with the preoperative score. There was no statistically significant inter-eye difference. Conclusion: Multiple studies of visual outcomes after traditional SLTS pituitary surgery have shown favorable results with this technique. The methods of visual field testing and visual field scoring in the current study are not directly comparable to previous studies of outcomes from traditional surgery. However, the current preliminary data suggest that visual outcomes after MIPS are favorable as well. This implies that MIPS is a reasonable alternative to tradition pituitary surgery.

 
 
 

Health Behaviors in Primary Care Patients at Risk for Diabetes

Xiaoci “Sarah” Guo, MSII, Katrina E Donahue1, MD, MPH, Madeline Mitchell2, MURP, Philip D. Sloane1,2, MD, MPH

1Department of Family Medicine, 2 Cecil G. Sheps Center for Health Services Research

University of North Carolina, Chapel Hill, NC
 
ABSTRACT
 

PURPOSE: Poor health behaviors contribute to developing diabetes. This study examines the relationship between patients’ risk for diabetes and their health behaviors, including nutrition, physical activity, smoking, and alcohol use to understand health habits of this population to effectively tailor office counseling.

 

METHODS: Patients presenting for routine visits at six NC family practices were asked to fill out a computer survey. Those identified as having one or more risky behavior were enrolled. Study participants received a follow up telephone survey to obtain additional information regarding their health behaviors. Health behaviors were compared among patients with 1) diabetes, 2) high risk for developing diabetes and 3) low risk.

 

RESULTS: Currently, 109 patients are enrolled. Patients with diabetes or at risk for diabetes are more likely to be smokers than those at low risk (28.6% vs 22% vs 16.1%). Patients with diabetes or at high risk are more likely to get less than 5 servings of fruits and vegetables per day than those at low risk (53.6% vs 68.0% vs 21.0%). Patients with diabetes or at risk for diabetes are more sedentary than those at low risk (42.9% vs 74% vs 22.6%).

 

CONCLUSIONS: Similar to patients with diabetes, those at risk for diabetes are smokers, have poor nutrition habits and are sedentary. For patients at risk for developing diabetes ‘5 fruits and vegetables a day’ appears to be one important nutrition counseling message. Additionally, small steps in increasing any type of activity will also help high risk patients.

 

The Role of the Serotonin 2A Receptor Subtype in Primary Auditory Cortex Layer II/III Pyramidal Neurons

Scott Daniels1, Gregory Basura2, MD, PhD, Deepti Rao3, Paul Manis4

1 School of Medicine MD Candidate, 2 Otolaryngology Resident, 3 Cell and Molecular Biology PhD Candidate, 4 Otolaryngology Department Chair

 

Supportedby the Otolaryngology Training Grant

University of North Carolina, Chapel Hill, NC

 

Background and Significance: The advent of Cochlear Implants for patients with Sensory Neural Hearing Loss (SNHL) caused by impaired cochlear function has revolutionized the treatment of this disorder. However, studies have shown that cochlear implants are not as effective as was originally hoped, especially in older patients, implying that there may be damaging alterations to the central neural auditory pathway components following SNHL. Gross changes have been noted, however, there may also be largely unexplored changes in individual neuron attributes. Serotonin (5-hydroxytriptamine, 5-HT) is known to actively participate in early cortical development. Beique et al. (2004) showed that during normal development of the prefrontal cortex, there is a coordinated shift in 5-HT receptor subtype expression, dramatically affecting on the excitability in response to identical serotonin stimulus. We gathered data concerning the serotonin pharmacology in normal rats, using serotonin and ketanserin, a 5-HT2A/C selective antagonist. Methods: Anesthetized rats (post-natal days 10-19) were decapitated and slices of living brain tissue 400 µm thick were made in a trajectory that includes the intact thalamacortical neural connections. Cell bodies in layers II/III were visualized with video-enhanced differential interference contrast technology and real time current clamp recordings were made. Drugs were administered by bath application at concentrations of 50μM and 1 μM for serotonin and ketanserin, respectively. Cells were monitored using a test-pulse alternating with a holding period without current injection. Results: Control cells treated with serotonin showed a marked and reversible decrease in input resistance and average number of action potentials per test-pulse interval. The decrease in average number of action potentials per test-pulse interval was blocked by ketanserin, but the decrease in input resistance was not. Other measured parameters did not vary in a statistically significant way. Further anatomical and electrophysiological data from cochlear ablated animals will be presented.

 

Comparative Accuracy of Oscillometric and Intra-aortic Blood Pressure Measurements In Patients Undergoing Routine Cardiac Catheterization.

Skand Bhatt1, Robert Kelly MD2, Rick Stouffer MD2

1School of Medicine and Public Health, 2Department of Cardiology

University of North Carolina at Chapel Hill

 

Oscillometry is a common method of non-invasive blood pressure determination, but few studies have reported the accuracy of this technique compared to direct measurement. To assess the accuracy of oscillometry, non-invasive cuff pressures were measured at the brachial artery and compared with simultaneous direct aortic pressure measurements in 98 patients (32-84 years, mean 58.5 years, 55.1% males) undergoing routine cardiac catheterization. Oscillometry underestimated mean aortic systolic pressure by 7.4 mm Hg (p<0.0001) and overestimated mean aortic diastolic pressure by 5.2 mm Hg (p<0.0001). Clinically significant discrepancies, defined as greater than +/- 10.0 mm Hg difference, were observed in 49.0% of mean systolic measurements and 30.6% of mean diastolic measurements. In a subgroup analysis, mean pressure differences were significantly greater for women compared to men after adjusting for other known factors. The results suggest that clinically significant differences in systolic and diastolic pressure exist by the oscillometric technique, compared to direct measurements, and that the magnitude of difference may be greater among female patients.

 
 

Regional Change in Brain Morphometry in Schizophrenia Associated with Short Term Antipsychotic Treatment.

Stacy Greeter, 1 Khary Carew, 1 Robert McClure, M.D. 2

UNC-Chapel Hill School of Medicine,1 Department of Psychiatry, UNC2

 
 

Previous studies have shown volume changes in basal ganglia and cerebral cortex of schizophrenia patients undergoing long-term treatment with typical and atypical antipsychotics. The aim of this study was to identify if caudate and cortical volume changes occur over short periods of time (3 months) in schizophrenia patients treated with typical and atypical antipsychotics. Patients satisfying DSM-IV criteria for schizophrenia were withdrawn from medication for three to six weeks. MRIs were performed after medication withdrawal and after 12 weeks of typical or atypical antipsychotic treatment. Region-of-interest (ROI) analysis was performed. Absolute volume changes measured by ROI were in the expected directions based on previous longitudinal studies. Caudate volume of typically treated patients increased and caudate volume of atypically treated patients decreased or did not change. The caudate results were not statistically significant, perhaps due to small sample size. Our results suggest that brief periods of treatment with antipsychotic medications are associated with changes in caudate volume.  

 

ROLE OF MACROPHAGE ACCUMULATION AND ALTERED SIGNALING IN INNATE IMMUNE DYSFUNCTION LATE AFTER BURN INJURY

 

Stefan Mlot BS1, Robert Maile PhD1,2, and Bruce Cairns MD1

The University of North Carolina at Chapel Hill

NC Jaycee Burn Center, Departments of 1Surgery and 2Microbiology and Immunology

 

BACKGROUND: Burn injury is known to induce an early (three days post-burn) suppression of the adaptive immune system, an early priming of the innate immune system, and a late (14 days post-burn) hyperresponsiveness of both the innate and adaptive immune systems. Activation of the innate immune response is mediated in part by stimulation of Toll-like receptors. Late after burn injury, macrophages have been shown to accumulate in the spleen, and splenocytes have been shown to exhibit increased apoptosis and cytokine synthesis upon Toll-like receptor (TLR) stimulation. AIMS: The goals of this project were 1) to characterize the splenic macrophage population in burned mice compared to sham 14 days after burn injury and 2) to determine the variation in expression of various TLRs in these macrophages. METHODS: Splenocytes were harvested from B6 mice 14 days after burn or sham injury. Anti-CD11b and anti-F4/80 antibodies were used to characterize splenic macrophages, and CD11bhiF4/80hi and CD11bhiF4/80int populations were sorted by flow cytometry. From the total RNA obtained from these populations, cDNA was synthesized and quantitative RT-PCR was performed to determine the fold change in expression of TLRs in burned mice compared to sham. RESULTS: We found that the accumulation of macrophages in the spleen can be attributed largely to an eight-fold expansion in CD11bhiF4/80int macrophages (p<0.001). However, expression of TLR2, TLR4, and TLR9 decreased or remained constant in both this population and a non-expanding CD11bhiF4/80hi population. DISCUSSION: These data indicate that there is a significant enrichment of macrophages in burn mice late after burn injury with significant reduction of the expression of TLR2, TLR4, and TLR9 in those same macrophages. This suggests that the enhanced cytokine response to TLR ligation found in the splenocyte population is due to the overpowering effect of macrophage accumulation in the spleen rather than increased TLR expression.

 

Heparin sodium salt does not promote collagen fibril sliding in rat tail tendons, in vitro.

Taylor Stone; Laury Dahners, M.D.

Department of Orthopaedics, University of North Carolina, Chapel Hill, NC

 

Despite advances in joint surgery and postoperative management, postoperative stiffness continues to be a relatively common phenomenon. For example, studies have reported stiffness in >50% of patients with total knee arthroplasty, although the true incidence appears to be 8% to 12%. In order to develop treatments for this surgical complication, it is necessary to learn more about the composition of tendons and ligaments and to exploit the relationships among the various extra-cellular components that allow growth and contracture. The sliding fibril hypothesis postulates that length changes in dense collagenous tissues occur primarily through sliding in discontinuous collagen fibrils past one another and that fibrils are reversibly connected through a stress-transfer matrix. In this experiment, heparin and a low-molecular-weight (LMW) heparin were tested as potentiators of tendon creep because of known interactions with extra-cellular molecules believed to function in fibril sliding. We suspended stained rat tail tendons under a constant stress and observed collagen fibril sliding as shown through length changes in the tendons. Although heparin is known to bind to type I collagen, decorin, and fibronectin, neither it nor LMW heparin demonstrated the ability to potentiate tendon creep secondary to collagen fiber sliding.   

 
 

Comparison of Functional MRI Activation Patterns between Mild Cognitive Impairment Subjects and Elderly Controls at Ultra-High Field Strength

Sriyesh Krishnan1,2, Melissa Slavin1,2,3, Thanh-Thu Tran1,2,3,Lakshmi Murty1,2, Anne Finefrock 3, P.Murali Doraiswamy3, Jeffrey Petrella1

1Department of Radiology, 2Brain Imaging and Analysis Center, and 3Department of Psychiatry

Duke University Medical Center, Durham, NC
 
 

BACKGROUND: The Mild Cognitive Impairment (MCI) population represents a group of at-risk subjects who may have prodromal Alzheimer’s disease (AD). Analysis of subtle functional brain activation changes in this population may yield information on patterns of neuronal dysfunction occurring prior to anatomical changes seen with conventional imaging techniques.    OBJECTIVE: To assess differences in brain activation patterns between MCI subjects and normal controls using functional magnetic resonance imaging (fMRI) at ultra high field strength during a memory encoding and retrieval task.    METHODS: Twenty-eight subjects were studied during a face-name memory encoding and retrieval task at 4T. Novel and familiar face-name pairs were presented within a blocked design. The MCI (9M/5F, mean 76.3 years, SD 6.1) and control (9M/5F, mean 72.7 years, SD 5.0) groups were compared using Statistical Parametric Mapping random effects ANCOVA, with age as the covariate. Medial temporal lobe (MTL) and whole brain analyses were performed.    RESULTS: Analysis during encoding revealed increased activation in the right precentral and fusiform gyri and left medial frontal gyrus in ONS subjects and increased MTL activation in the left hippocampus and parahippocampal gyrus in MCI subjects.   During retrieval, there were multiple areas of increased frontal activation in ONS subjects including bilateral insular cortex and left precentral gyrus. A single cluster of increased activation in MCI subjects in the area of the left fusiform and middle occipital gyri was noted during retrieval.    CONCLUSIONS:Differential fMRI activation patterns within MTL and frontal lobe structures are evident in MCIs in comparison to controls. These differences may reflect early disruption of the neuronal networks of memory processing prior to neuroanatomical atrophy. Analysis of differential fMRI activation could someday have a diagnostic role in the early detection of AD.

 
 
 

Effects of Couples Intervention on the Experience of Fatigue

in Breast Cancer Patients
 

Theresa Kallman1, Donald Baucom2,3, Tina Gremore2,3, Nicole Pukay-Martin2,3

1 School of Medicine 2 Department of Psychology 3 Lineberger Comprehensive Cancer Center

University of North Carolina, Chapel Hill, NC
 

Fatigue is experienced by 60 to 90 percent of cancer patients, and thus represents a significant barrier to achieving a high quality of life. Decreasing the physical side effects of breast cancer, in this case, fatigue, is an obtainable goal in the treatment of breast cancer patients. Given the link between psychological factors and fatigue, psychosocial interventions for breast cancer patients are likely to impact the patient’s experience of disease positively. The goal of the current investigation is to understand the mechanisms through which the CanThrive intervention contributes to lower levels of fatigue for women with breast cancer. Measures of pain, fatigue, social support, marital satisfaction, psychological functioning, and cancer related well-being are assessed in the CanThrive study as part of the routine research assessment battery. Baron and Kenny’s approach to assessing mediation was employed to examine potential mechanisms through which CanThrive affects fatigue. It was found that the CanThrive intervention decreases fatigue and also decreases psychological distress. It was then found that psychological distress accounts for partial mediation of the relationship between treatment group and fatigue. That is, CanThrive appears to decrease fatigue in part by decreasing psychological distress. This study found that the CanThrive intervention decreases fatigue, even though fatigue is not directly targeted. This is important because even though fatigue is medically induced, behavioral and psychosocial strategies can be effective additions to medical interventions for decreasing the experience of fatigue among women with breast cancer. The results of this study show that the effect of CanThrive is not medicated by social support or relationship variables such as relationship adjustment, but rather is a result of decreasing individual psychological distress. Moreover, CanThrive decreases women’s psychological distress which is related to their experience of fatigue.  

 
 

Proyecto Puentes de Salud: An Exploration of Cardiovascular Health and Risk Factors in Rural Mexico

Allan D Nanney III5, Trista D Snyder5, Ian J Nelligan5, Amanda L

Rollins5, Katherine Zeman, Michael Clarke-Pearson, Pamela Y Frasier1

PhD MA MSPH, Bron Skinner1 PhD, Daniel Reuland3 MD,

Flavio Rojas4 PhD, Mauricio G Cohen2 MD.

1UNC Department of Family Medicine, 2UNC Department of Cardiology, 3UNC Department of Medicine, 4UNC Department of Biostatistics, 5UNC School of Medicine

University of North Carolina, Chapel Hill, NC

 
 

The 2000 NC census estimated a 1200% increase in Hispanic immigration to the Triangle area. A large proportion of the economic migrants come from rural Mexican communities, and little is known about their antecedent lifestyles and health risks and how these might contribute to their current health status. Therefore, Proyecto Puentes de Salud was initiated to determine cardiovascular risk factors of rural residents in Guanajuato, Mexico. Future studies will draw correlations between health and lifestyle risks of the US immigrant and rural Mexican populations, providing NC physicians with an all-encompassing view of Latino health. A convenience sample of 267 participants, recruited from six rural communities in Guanajuato during June and July of 2006, were (1) offered free screenings for cholesterol, fasting glucose, blood pressure, and obesity; (2) counseled about risk factors for cardiovascular diseases and diabetes; and (3) interviewed using a standardized questionnaire that included demographic information, past medical history, family history, dietary and lifestyle behaviors, and social and psychological health.267 participants (ages 20-85) were screened and interviewed. Prevalence of hypertension (>140/90) was 29.96%, impaired fasting glucose (>100 mg/dL) was 25.02%. Frequency of abnormal cholesterol and HDL was 19.85 % (>200 mg/dL) and 85.55% (<40 mg/dL men, <50mg/dL women), respectively. Abdominal obesity rates were 73.03%. We detected a high prevalence of cardiovascular risk factors in rural Mexicans that warrants further investigation. In particular, we noticed an overwhelming presence of inadequate levels of HDL and abdominal obesity, placing this population at risk for The Metabolic Syndrome. Lack of education on cardiovascular risk factors and healthy lifestyles seem to play a large role in perpetuating poor diet, limited exercise, and daily health choices. Additional educational instruction, with lifestyle modifications could rectify these risks; improving the health of Mexicans in their place of origin may have a ripple effect on the health of Latino immigrants we treat in our community. Also, understanding of this expanding patient population will give American physicians a head-start when treating and educating in the clinic. The study was limited by the under-representation of men in the research and by being a convenience sample.