BIRCWH Poster Abstracts

1 Department of Cancer Biology, Vanderbilt University Medical Center 2 Vanderbilt Center for Bone Biology, Vanderbilt University Medical Center P-1 Osteoblasts at Bone Site Promote Osteolytic Bone Metastasis

Background and Objective: Metastatic breast cancers are often associated with osteoclastic bone resorption, which releases active TGFβ in the bone. Increased TGFβ in the bone suppresses osteoblast differentiation. We have demonstrated that undifferentiated stromal cells (C2C12) secrete significantly higher TGFβ compared to committed osteoblasts (2T3). We hypothesize osteoblast-derived TGFβ may promote bone metastasis. Our objective was to investigate whether osteoblasts directly increase metastatic behavior of breast cancer cells to promote bone metastasis.

Methods: We have injected MDA-MB-231-GFP human breast cancer cells in female nude mice using intracardiac, intratibial, or subcutaneous models to assess bone metastasis and to test in vivo motility.

Results: Anti-TGFβ antibody 1D11 increased bone volume in addition to reducing tumor burden in the intracardiac model, suggesting an increase in osteoblast differentiation. Conditioned media from undifferentiated stromal cells (C2C12) increased motility (P < 0.001) of MDA-MB-231-GFP cells in a TGFβ-dependent fashion compared to committed osteoblasts (2T3). Subcutaneously injected MDA-MB-231 cells migrated toward contralaterally injected MG63 (secretes 125 pg/ml/106 cells TGFβ). When co-injected intratibially with MDA-MB-231 cells, MG63 cells significantly (P < 0.05) increased osteolytic lesion area compared with 2T3 cells (lesion area: 12,000 arbitrary units in MG63 versus 3,000 arbitrary units in 2T3). This increase in osteolysis may be due to the increased amount of secreted TGFβ from MG63.

Conclusions: Osteoblasts-derived TGFβ may influence motility and osteolysis of the breast cancer cells. Our future direction is to further investigate the role of undifferentiated osteoblasts in metastatic breast cancer to the bone.

1 Division of Surgical Oncology, Department of Surgery, Virginia Commonwealth University and Massey Cancer Center 2 Department of Biochemistry and Molecular Biology, Virginia Commonwealth University and Massey Cancer Center P-2 Estradiol-Mediated Export of Sphingosine 1-Phosphate from Breast Cancer Cell Is Via Non-Genomic Effect of Estrogen Receptor and Not GPR30

Background and Objective: Sphingosine 1-phosphate (S1P) is an active lipid mediator that plays critical roles in breast cancer progression by causing cell proliferation, migration, and angiogenesis. S1P exerts its functions by binding to its specific receptors on the cell surface after it is intracellularly produced by sphingosine kinase 1 (SphK1). Recently, estradiol (E2), which binds to estrogen receptor (ER) or GPR30, has been identified as one of the stimulators of SphK1. Our objective is to elucidate how E2 mediates S1P export from breast cancer.

Methods: ER positive MCF-7 and negative MDA-MB-231 human breast cancer cells were used as models. Eighteen hours of incubation with ICI182780 were used to inhibit ER expression, and its short-term incubation and G-1 compound were used to stimulate GPR30. S1P was measured using 3H-labeled sphingosine, or mass spectrometer (LC-ESI-MS/MS).

Results: E2, but not EGF, phosphorylated SphK1 and released S1P hours earlier than SphK1 transcription was induced, although both produced S1P in the cells. Inhibition of ER using ICI182780 blocked E2-mediated S1P export, but not the baseline secretion. However, MDA-MB-231 cells did not export S1P at all. Further, stimulation of GPR30 using either short-time ICI182780 or G-1 did not induce S1P export.

Conclusions: This is the first demonstration that non-genomic effect of E2 on S1P export was mediated via ER and not GPR30. Since it has been clearly shown that S1P is involved in breast cancer survival, these findings may, at least in part, explain why ER positive breast cancers are less responsive to initial chemotherapy.

1 Department of Obstetrics and Gynecology, David Geffen School of Medicine 2 Department of Microbiology, Immunology and Molecular Medicine, David Geffen School of Medicine 3 Department of Molecular and Medical Pharmacology, David Geffen School of Medicine 4 Howard Hughes Medical Institute, University of California, Los Angeles 5 Department of Obstetrics and Gynecology, Feinberg School of Medicine, Northwestern University P-4 Establishment of an In Vivo Mouse Model for Endometrial Regeneration from Dissociated Populations of Endometrial Stroma and Epithelium

Background and Objective: Endometrial cancer is the most common gynecologic cancer in the United States. Understanding genetic mechanisms and the target cell population that can initiate endometrial carcinoma can help identify novel targeted therapeutics. A major obstacle has been lack of reliable in vivo mouse models that can accurately recapitulate human disease. Our objective is to establish an in vivo mouse model for regeneration of endometrial glands that can be used to study oncogenic transformation and isolate putative stem cells of endometrial glands.

Methods: An endometrial tissue recombination system has been established previously (Cunha 1976). In this model, endometrial glandular and stromal tissue fragments are recombined and regenerated into endometrial glands under the kidney capsule of a severe combined immunodeficiency (SCID) female mouse (Cunha 1976). To genetically manipulate endometrial stroma and epithelia independently with retro or lentiviral delivery vectors, we have established a modified version of this tissue recombination system. In our model, endometrial stroma and endometrial glands are dissociated into single cells, allowing genetic manipulation of each compartment. Dissociated stromal and epithelial cells are placed into a collagen plug and implanted under the kidney capsule of a SCID mouse for 6 to 8 weeks.

Results: Regenerated grafts in this model result in the formation of endometrial glandular-like structures closely resembling the native murine endometrium based on morphology and marker expression profile.

Conclusions: We have established a powerful in vivo regeneration system for murine endometrial cells that can be used to (1) study malignant transformation and (2) isolate putative stem cells (potential target cells for endometrial cancer).

1 University of Illinois at Chicago 2 University of Michigan P-5 Young Women Diagnosed with Cancer: A Preliminary Description of Their Decision-Making Process Surrounding Fertility Preservation

Background and Objective: Advances in cancer therapy have dramatically lengthened survivorship for young women. Concurrently, breakthroughs in reproductive technologies have recently resulted in advanced fertility preservation treatment such as oocyte, embryo, and ovarian cryopreservation. This study focuses on the decision-making process of young women who are diagnosed with cancer and are confronted with the choice of undergoing or foregoing advanced fertility preservation treatment.

Methods: Women were recruited from two large research institutions and the Internet. All participants were newly diagnosed with cancer, were medically eligible for fertility preservation treatment, and had discussed treatment options with their physicians. A qualitative, naturalistic approach guided the analysis.

Results: The decision-making process of young women consists of six interrelated phases: absorbing the cancer diagnosis, accepting diminishing fertility, contemplating treatment, engaging in self-examination, selecting a course of action, and reflecting upon the decision. Within these phases, women responded to their cancer diagnosis and envisioned how an abbreviated fertility period may affect their lives. They contemplated fertility preservation treatment by applying their values about motherhood while weighing cost and perceived risk. Women experienced a period of introspection and ultimately reached a decision about whether to undergo treatment. Finally, the women reflected upon the meaning of their decision.

Conclusions: We identified six phases that young women progress through when making the decision to undergo or forgo fertility preservation treatment. Healthcare providers can optimize decision support by identifying where patients are in the process and providing phase-specific counseling.

1 Department of Surgical Oncology, Virginia Commonwealth University Health System 2 Department of Medical Oncology, Virginia Commonwealth University Health System P-7 How Should Women with Lobular Neoplasia on Core Biopsy Be Treated? A Literature Review

Background and Objective: Lobular neoplasia (LN) of the breast comprises lobular carcinoma in situ (LCIS) and atypical lobular hyperplasia (ALH). Traditionally a marker of an increased risk of breast cancer, they may be a preneoplastic lesion. Their management is controversial when identified on core needle biopsy (CNB) secondary to the risk of a synchronous and/or the development of a metachronous breast malignancy. Around 36,000 women each year have to decide on treatment options ranging from careful followup to mastectomy. A literature review was performed to assess if clinical or biologic markers would provide risk stratification for the presence of a synchronous or the development of a metachronous breast cancer for women with LN on CNB.

Methods: Pub Med was searched using “atypical lobular hyperplasia breast” and “lobular carcinoma in situ breast.” The retrieved abstracts were reviewed and the pertinent articles read, along with any key references from these articles.

Results: Multiple clinical and pathologic features have been evaluated for their predictive value. None consistently stratifies the risk for a synchronous and/or metachronous breast cancer for women with LN on CNB.

Conclusions: The significance and treatment of LN of the breast is controversial. There are no clear features of LN that define the risk of a synchronous and/or metachronous breast cancer. Research in invasive breast malignancies has identified biological makers that predict recurrence and direct treatment. Biological changes in lobular neoplasias and their correlation with clinical characteristics may be useful in defining treatment and significance.

1 Division of Radiobiology, Department of Radiation Oncology, Virginia Commonwealth University 2 Division of Surgical Pathology, Department of Pathology, Virginia Commonwealth University P-8 Reciprocal Interactions between Extracellular Matrix Signaling and DNA Double-Strand Break Repair in Breast Cells

Background and Objective: Genomic instability and acquisition of invasiveness through extracellular matrix (ECM) accompany breast cancers. Using human breast cell lines HMT-3522 with non-invasive S1, pre-invasive S3-C, and invasive T4-2 counterparts, we found that (1) ECM signaling via β1 integrin regulates DNA double-strand break repair (DSBR) pathway homologous recombination (HR), and (2) some repair proteins, such as polo-like kinase 1 PLK1, are involved ECM invasion, designating these GISEM for Genome Instability and Extracellular Matrix invasion. Our objective is to decipher how ECM signaling regulates DSBR and, reciprocally, how DSBR genes function in invasion through ECM.

Methods: siRNA inhibition, chemical inhibitors, and blocking antibodies were used in HMT-3522 cells. We used a GFP-based assay for HR, transwell assays for invasion through Matrigel^®, and immunohistochemistry for patient tissues.

Results: (1) In S1 cells, we found that the β1 integrin specific interactor ICAP1 is a negative mediator of ECM-to-HR signaling. Downstream ROCK1 is involved in mediating ECM signals to HR. The adherens junction protein E-cadherin, which is also upstream of ROCK1, regulates HR and mediates nuclear levels of RAD51, γ-H2AX, and beta-catenin. (2) DSBR protein MRE11/RAD50/NBS1 (MRN) levels are upregulated by ECM in S1 cells but not in S3-C and T4-2. MRN protein levels decrease in progression from benign to pre-invasive to invasive lesions in 48 patients. (3) The DSBR protein XRCC3 negatively regulates invasion in T4-2 cells. XRCC3 is downregulated in progression from S1 to S3-C to T4-2.

Conclusions: We deciphered new interactions between ECM and DSBR with potential significance in breast cancer.

1 Department of Family Medicine, University of Maryland School of Medicine 2 School of Pharmacy, University of Maryland 3 Pennsylvania State University P-9 Terameprocol Efficacy Studies in HPV Infection Models: A Candidate Chemoprevention Agent in Cervical Cancer

Background and Objective: Tetra-O-Methyl Nordihydroguaiaretic Acid (terameprocol) has Sp1 promoter inhibition properties and anti-HPV effects via inhibition of the p97 promoter and anti-cancer effects via Cdc2 inhibition and G2/M arrest of dividing cells. Terameprocol is formulated as a vaginal ointment. Our objective is to demonstrate the efficacy of topical terameprocol against HPV in vitro and in vivo using the CRPV/rabbit model.

Methods: In vitro: A431 cells were plated at 2 × 10⁵ cells/well in six-well cluster dishes. Replicate aliquots of HPV-11 were added to each representing an MOI of 150 particles/cell. Titrations were added 48 hours after addition of virus and incubated a further 72 hours. Cell cultures were harvested, lysed with Trizol reagent, and RNA prepared. Quantitative real-time polymerase chain reaction (PCR) was conducted to quantify the proportion of viral E1^E4 transcripts (EC50) and a cellular reference RNA for the TATA-binding protein (CC50). In vivo: The CRPV rabbit model was used to determine efficacy of terameprocol 2% ointment and 2% terameprocol in DMSO. We induced skin papillomas using purified mE8-CRPV on scarified rabbit skin. Dosing regimes include 1% and 2% ointment, 2% solution in DMSO, and controls with white petroleum and DMSO.

Results: In vitro: We found good efficacy of terameprocol against HPV-11 infection (EC50 of 0.4 μg/ml, CC50 of 5.0 μg/ml, Selectivity Index of 12.5). In vivo CRPV/rabbit study: Terameprocol showed suppression of papilloma growth for group B (2% in petrolatum).

Conclusions: Terameprocol demonstrates encouraging anti-HPV activity in vitro and effectiveness at reducing CRPV-induced papillomas, with some cures observed. Other treatment groups showed no statistically significant reduction.

1 University of Maryland School of Medicine 2 Department of Physical Therapy, University of Delaware 3 National Institute on Aging P-10 Sex Differences in the Relationship of Serum Carotenoid Levels with Mortality Risk among Older U.S. Adults: The Third National Health and Nutrition Examination Survey (NHANES III)

Background and Objective: Serum carotenoid levels are the best measure for diets rich in fruits and vegetables. No study has examined sex differences in the association of serum carotenoids with mortality in a representative sample of older U.S. adults. We aim to examine whether low serum carotenoid levels relate to mortality risk among older adults and to determine whether the association differs by sex.

Methods: The sample included 2,683 adults (1,315 men, 1,368 women) ages 65 years and older from NHANES III. Weighted Cox models assessed the relationship between total serum carotenoids and all-cause mortality. Total serum carotenoids were the sum of serum beta-carotene, alpha-carotene, beta-cryptoxanthin, lycopene, lutein, and zeaxanthin.

Results: The median total serum carotenoid level was 1.43 μmol/L (interquartile range: 1.00, 1.93 μmol/L). Women had higher median total serum carotenoids than men (1.55 v. 1.25 μmol/L, p < 0.001). Women had 455 deaths in 123,301 person-years; men had 643 deaths in 106,530 person-years. After adjusting for covariates, men in the lowest carotenoid quartile had a 17% higher all-cause mortality rate than men in the upper three carotenoid quartiles (incidence rate ratio [IRR] 1.17, 95% confidence interval [CI] 0.96–1.43, p = 0.13); and women in the lowest carotenoid quartile had a 53% higher all-cause mortality rate than women in the upper three carotenoid quartiles (IRR 1.53, 95% CI 1.16–2.02, p = 0.002). The IRRs were not significantly different (p-value for interaction = 0.12).

Conclusions: The findings show a strong association between low total serum carotenoid levels and all-cause mortality in women, but not men, among a representative sample of older U.S. adults.

1 Department of Obstetrics and Gynecology, Duke University, San Francisco 2 Department of Biostatistics, Duke University, San Francisco 3 Department of Obstetrics and Gynecology, University of California, San Francisco P-11 Prevalence and Incidence of Urinary Incontinence in a Longitudinal Study of a Diverse Cohort of Women with Benign Gynecologic Conditions

Background and Objective: The proportion of women presenting with bleeding, pelvic pain, and fibroids, who also have urinary incontinence (UI) is not clear. Our objective is to determine the prevalence and incidence of UI in a diverse cohort of women with gynecologic conditions who participated in a 4-year longitudinal study and to assess factors associated with UI prevalence and incidence.

Methods: We conducted a secondary data analysis of SOPHIA (Study of Pelvic Problems, Hysterectomy, and Intervention Alternatives), a longitudinal study of women with benign gynecologic conditions (bleeding, pelvic pain, and fibroids). Urinary incontinence was defined as UI in the last 4 weeks, based on interviewer-administered annual questionnaires. UI type (stress [SUI], urge [UUI], or mixed incontinence [MUI]) was also evaluated.

Results: 907 women were included: 40.8% White, 28.0% Black, 17.3% Latina, and 8.1% Asian. The mean age was 44.1 ± 5.4 years. The overall prevalence of any UI was 51.1%. At baseline, SUI was most common (39.4%), followed by UUI (23.7%), and MUI (18.9%). The average annual incidence for any UI was 4.2%: 4.0% SUI, 3.5% UUI, and 3.4% MUI. Prevalent UI was associated with age in decades (Odds Ratio [OR] 1.4, 95% CI 1.1–1.8), parity (OR 1.8, 95% CI 1.3–2.3) and ethnicity (Latina compared to White; OR 2.0, 95% CI 1.3–3.0). None of the factors was associated with incidence of UI.

Conclusions: In a diverse population of women with benign gynecologic conditions, over half of the women have urinary incontinence. Providers should ask about UI symptoms when treating patients with benign uterine conditions.

1 Department of Physiology, Tulane University School of Medicine, New Orleans 2 Hypertension and Renal Center, Tulane University School of Medicine, New Orleans 3 Tulane-BIRCWH Program, Tulane University School of Medicine, New Orleans P-12 Effect of High-Salt Diet on Renin and Prorenin Receptors in Kidney Distal Nephron Segments May Help To Explain Sex-Dependent Differences in the Development and Progression of High Blood Pressure and Renal Injury

Background and Objective: Hypertension and kidney injury are more severe in men than in women. Collecting duct (CD) renin and prorenin receptors [(P)RR], components of the renin-angiotensin system, play a role in AngII-dependent hypertension. Our objective is to determine if the effect of a high salt diet (HS) on CD renin and (P)RR gene expression contributes to sex-dependent differences in systolic blood pressure (SBP) and proteinuria in chronic AngII-infused male (n = 23) and female (n = 22) rats.

Results: Baseline SBP did not differ between males and females (116 ± 3 v. 119 ± 2 mmHg) or after AngII infusions (228 ± 9 v. 217 ± 20 mmHg). HS added to AngII-infusion did not modify SBP (228 ± 13 v. 211 ± 16 mmHg); however, it increased proteinuria to a greater extent in male than females (79 ± 6 v. 47 ± 6 mg/day). CD renin-mRNA levels were higher in males than females (1.0 ± 0.4 v. 0.2 ± 0.06 au). HS increased renin-mRNA in females but not in males (1.1 ± 0.1 v. 2.5 ± 0.4 au). Renin protein levels were similar in male and female rats, but prorenin levels were higher only in females. HS increased prorenin only in female rats (0.4 ± 0.3 v. 3.1 ±0.4 au). Male and female rats exhibited similar increased renin gene expression during AngII infusions. Although baseline (P)RR-mRNA levels were twice higher in female than male rats, HS down-regulated the (P)RR-mRNA only in females (male [controls = 1.7 ± 0.1 v. HS = 1.9 ± 0.1], female [controls = 3.2 ± 0.4 v. HS = 2.1 ± 0.2]).

Conclusions: HS does not exert additive effect on SBP nor contribute to SBP sexual dimorphism. Augmented proteinuria in males at SBP comparable to females may reflect a greater degree of renal injury, which might be explained by the increased CD renin and (P)RR gene expression. During HS, (P)RR down-regulation in female rats may represent a protective mechanism against kidney injury in this gender.

1 Section of Gastroenterology and Hepatology, University of Wisconsin School of Medicine and Public Health 2 Center for Women's Digestive Disorders, Women and Infants Hospital, Alpert Medical School of Brown University P-13 Creation of a Women's Health Training Program in Gastroenterology

Background and Objective: Women's health tracks have been developed to address deficiencies in women's health education. Likewise, women's health competencies have been integrated into training programs. However, knowledge and confidence in evaluating and managing women's health issues remain low. In gastroenterology, nearly 60% of patients are female. Training in gastrointestinal (GI) women's health is, therefore, critical. Our objective is to (1) create a GI women's health training experience and (2) develop academic gastroenterologists focused on GI women's health.

Methods: (1) A multidisciplinary group of GI women's health experts was formed; (2) focus groups with primary care providers and sub-specialists were held to determine unmet needs in the management of the GI health of female patients; (3) a syllabus was developed for self-directed learning; (4) rotation in GI women's health was introduced for trainees in general gastroenterology; and (5) GI women's health track was introduced for trainees committed to academic careers.

Results: Since 2002, 13 fellows who participated in the rotation have graduated from the Brown GI fellowship program. One fellow has completed the track. Satisfaction with the program is high. All graduates participating in the rotation for a minimum of 1 to 2 months rated their training in GI women's health “just right.” Participating graduates also felt prepared to manage pregnancy and other GI women's health issues. The graduate of the track is a clinician-researcher at an academic center.

Conclusions: A training experience in GI women's health can be created using local resources and expertise. Gastroenterologists with this training feel prepared to evaluate and manage the spectrum of women's health issues encountered in practice.

1 Department of Surgery, Tulane University School of Medicine 2 Department of Community Health Sciences, Tulane University School of Public Health and Tropical Medicine 3 Department of Medicine, Tulane University School of Medicine 4 Tulane Abdominal Transplant Institute 5 Department of Epidemiology, Tulane University School of Public Health and Tropical Medicine P-15 Risk of Graft Loss Lowered for Female Recipients of Male Kidneys When Donors Are Larger

Background and Objective: Female recipients of male donor kidneys compared to all other gender combinations may have the worst graft survival, and patients who receive larger kidneys relative to their body size may have a graft survival advantage. We speculated that the inferior graft survival for female recipients of male kidneys may be improved if kidneys are from larger, male donors.

Methods: We studied 668 kidney transplants retrospectively. Univariate correlations were calculated using Pearson's correlation coefficient, and 3-year graft survival was determined by Kaplan-Meier estimation. Multivariate methods were performed to determine the hazards of graft loss.

Results: We identified 146 male to female transplants. This group compared to all others had the lowest graft survival (86%, 79%, and 78% v. 92%, 88%, and 86% at 1, 2, and 3 years, respectively, log-rank p = 0.01). Donor body mass index (BMI) and kidney surface area directly correlated (r = 0.396, p <0001). Multivariate analysis showed that female recipients of male kidneys had a significant risk of graft loss (HR = 2.86, 95% CI 1.25–6.58, p = 0.01]). Male donors with a BMI two units larger than that of the female recipient resulted in a significantly lower risk of graft loss (HR = 0.19, 95% CI 0.05–0.67, p = 0.01).

Conclusions: The reduction in graft survival experienced by female recipients of male kidneys appears attenuated when grafts are from larger, male donors. Because kidney size correlated with BMI, this beneficial effect is likely due to greater nephron mass provided by larger, male donors.

1 Department of Radiological Sciences, Washington University School of Medicine 2 Department of Radiation Oncology, Washington University School of Medicine 3 Division of Hematology and Oncology, University of California, Davis Cancer Center 4 Molecular Imaging Center, Washington University School of Medicine P-16 Imaging Sites of Metastasis Before They Happen: Targeting the Premetastatic Niche

Background and Objective: Bone marrow-derived hematopoietic progenitor cells (HPCs) that express the integrin a4β1 arrive at future sites of metastasis and form a receptive environment for tumor cells known as the premetastatic niche. The purpose of this project is to target a4β1 as an in vivo biomarker of the premetastatic niche using the high affinity ligand 64Cu-CB-TE2A-LLP2A and positron emission tomography (PET).

Methods: In this preliminary study, a4β1-negative MDA-MB-231/luc human breast tumor cells were implanted intra-arterially into mice. Metastasis developed spontaneously in multiple sites. The mice were monitored over a period of 30 days post-implantation by a combined bioluminescence (BLI)/microPET/microCT imaging study to determine whether increases in PET signal with 64Cu-CB-TE2A-LLP2A were observed prior to the development of luc+tumors in bone.

Results: Cumulative standardized uptake value data from an experiment with eight mice imaged on days −3 (control), 4, 10, 16, and 30 post-tumor cell injection show increased uptake of 64Cu-CB-TE2A-LLP2A over 30 days (p < 0.0001) at sites of bony metastasis. Preliminary histology and BLI data validate the influx of tumor cells at sites of PET signal.

Conclusions: These results suggest that PET imaging of the premetastatic niche is feasible. Future work will include (1) determination of the timing of HPC and tumor cell migration in mouse models of metastasis; (2) analysis of a4β1 expression in HPCs isolated from total marrow in tumor-bearing and tumor naïve mice; and (3) visualization of changes in the pattern of metastatic disease using circulating factors from primary tumor cells.

1 Department of Psychiatry, University of Maryland School of Medicine 2 Department of Physiology, University of Maryland School of Medicine P-17 The Role of Androgens in the Gender Difference of Major Depression: An Animal Model

Background and Objective: Women are twice as likely as men to suffer from major depression. Evidence indicates that androgens protect men from depression and contribute to this gender difference. The depression prevalence is similar between boys and girls. The gender difference in depression emerges in adolescence, paralleling the androgen surge in adolescent males. This has led to a general assumption that it is the adolescent androgen surge that underlies the gender difference in depression. However, little research has been done to explore the effects of perinatal androgen on depression. We want to determine whether perinatal androgens are antidepressant and whether the antidepressant effects of androgens associate with hippocampal neurogenesis.

Methods: Male pups were treated with vehicle or flutamide (androgen receptor antagonist) on post-natal day (PND) 0 and 1 to block neonatal androgen action. The depression-like behaviors were determined using the Forced Swimming Test (FST) and the Sucrose Preference Test (SPT) in preadolescent male rats. Doublecortin+neurons in the dentate gyrus, and the dendritic spines of pyramidal neurons in the CA1 region of the hippocampus were determined using immunohistochemistry and the Golgi-Cox technique, respectively.

Results: Neonatal-flutamide treatment caused not only a significant increase in depression-like behaviors, but also a decrease in doublecortin+neurons in the DG region and a decrease in the density of dendritic spines of pyramidal neurons in the CA1 region in preadolescent male rats.

Conclusions: Neonatal androgens may play an important role in protecting male rats from depression-like behaviors and promoting hippocampal neurogenesis in preadolescent male rats.

1 Department of Psychology, Pennsylvania State University 2 Indiana University P-18 Recognition of Wives' Emotional Expressions: A Mechanism in the Relationship Between Psychopathology and Intimate Partner Violence Perpetration

Background and Objective: Each year, approximately 14% of U.S. couples experience male-to-female intimate partner violence (IPV), frequently resulting in physical injuries and serious physical and psychological health consequences for women. The study of violent husbands' emotion recognition deficits is promising in terms of its theoretical base and implications for psychological therapies. We examine whether husbands' emotion recognition deficits serve as a mechanism in the relationship between two dimensions of psychopathology that commonly describe violent husbands and their IPV perpetration.

Methods: Among 88 community couples, both partners reported on male-to-female IPV. Husbands completed self-report measures of psychopathology. Wives were trained and photographed making facial expressions of emotions. Husbands completed an emotion recognition task including photographs of their wives and unfamiliar men and women displaying similar expressions.

Results: Husbands' diminished sensitivity to their wives' expressions of happiness partially mediated the relationship between their borderline/dysphoric characteristics and IPV perpetration. These relationships were specific to expressions of happiness displayed by their wives. In addition, husbands' IPV was associated with their diminished sensitivity to expressions of fear and their psychopathic personality characteristics were associated with misidentifying fearful expressions as neutral. However, the strength of husbands' sensitivity to fear as a mediator of the psychopathy-IPV relationship was suboptimal.

Conclusions: Study results support theories of IPV perpetration, demonstrate the significance of the intimate relationship, and question the applicability of a theory of extramarital violence to IPV. Implications exist for partner-specific emotion recognition training in therapies to prevent husbands' use of IPV, leading to improved health outcomes for women.

1 Department of Psychiatry and Behavioral Sciences, Medical University of South Carolina 2 Department of Neurosciences, Medical University of South Carolina P-21 Regional Areas of Brain Activation During Attempts To Resist Craving in Nicotine-Dependent Smokers: Preliminary fMRI Findings

Background and Objective: A strong association between craving and relapse to smoking during quit attempts among nicotine-dependent smokers has been demonstrated. Few studies have investigated the underlying neurological substrates during attempts to resist craving. We studied areas of regional brain activation during fMRI cue-induced craving and efforts to resist craving.

Methods: Participants (N = 4, F = 3) represent a pilot group of a larger sample of smokers participating in several fMRI craving studies. Participants were healthy, right-handed, non-treatment-seeking, nicotine-dependent adult smokers without major psychopathology and clinically significant medical problems. Two identical block design studies of fMRI cue-induced craving were completed after 2 hours of abstinence. During the first scan, participants were instructed to allow themselves to crave during cue presentation. During the second scan the instructions were varied to resist craving during the presentation of the smoking cues.

Results: Global brain activation decreased between craving and resist scans in 75% of the participants. The scans obtained during attempts to resist craving demonstrated increased right insula and temporal lobe activity compared to the craving scans. Craving scans showed more activity in bilateral middle prefrontal, middle cingulum, caudate nucleus, orbital frontal, and thalamus than resist scans.

Conclusions: Craving and cognitive strategies to resist craving utilize different regions of the brain. Recently, the amygdala has been implicated as a critical brain region involved in cigarette addiction. The insula has been demonstrated to have a potential role in the emotional and cognitive processes that promote drug use and may also be associated with craving resistance.

1 Department of Gerontology, University of Kentucky 2 Department of Psychiatry, University of Kentucky 3 Department of Behavioral Science, University of Kentucky P-24 The Feasibility of a Mental Health Treatment Engagement Intervention Program in Men and Women

Background and Objective: Mental health disorders are one of the leading causes of disability in women; however, many women do not get necessary treatment. This study is designed to expand the applicability of disease management and motivational interviewing into psychiatric appointment management, to prevent the under-treatment of psychopathology due to failure of treatment engagement. Research has indicated that mail and phone reminders, or appointment scheduling assistance, without motivational components do not effectively improve psychiatric treatment rates. Therefore, this project aims to conduct a trial examining the feasibility of a telephone-based brief motivational intervention as a method for increasing psychiatric treatment rates among both men and women (effectiveness has been seen earlier in men only).

Methods: The study has recruited N = 30 patients (50% women; 37% with depression as chief complaint) with intake appointments at the University of Kentucky Outpatient Psychiatry. Recruited patients are asked to complete baseline and 6-month followup telephone interviews. Patients randomized to receive the intervention participated in a brief telephone motivational interview targeting their barriers and motivation to attending their upcoming psychiatric intake appointment.

Results: Outcomes indicate 92% in the intervention group compared to 57% in the control group attended their intake psychiatric appointment, showing intervention effectiveness (p = 0.037) using chi-square analyses, with no interaction effect for gender.

Conclusions: Results indicate that the proposed intervention is feasible, acceptable, and effective for both men and women within an outpatient psychiatric unit. Engagement in treatment is the first step in ensuring treatment, consequently impacting psychiatric morbidity and mortality.

1 Department of Medicine, Brigham and Women's Hospital 2 Department of Psychiatry, Brigham and Women's Hospital 3 Department of Radiology, Brigham and Women's Hospital 4 Department of Medicine, Massachusetts General Hospital 5 Department of Psychiatry, Massachusetts General Hospital 6 Harvard Medical School, Massachusetts Institute of Technology 7 Department of Brain and Cognitive Sciences, Massachusetts Institute of Technology P-27 Stress-Response Circuitry Hypoactivity and Hormonal Dysregulation in Women with Depression

Background and Objective: Major depressive disorder (MDD) is a leading contributor to overall disease burden and is more likely to occur among women than men. Gender differences have been consistently observed, yet remain largely unexplained. Previous research on MDD suggests high rates of endocrine dysfunction and deficits in brain activation in the stress response circuitry (amygdala, hypothalamus, hippocampus, anterior cingulate gyrus [ACC], and orbitofrontal cortex [OFC]). This study was designed to investigate the relationship between HPA- and HPG-axis hormones, and stress response circuitry function in MDD.

Methods: During two points in their menstrual cycles (early follicular [EF], ovulation [OV]), female participants (10 with recurrent MDD, 10 controls) were scanned while viewing IAPS pictures on a 3T GE MR scanner. Baseline hormone measures included pituitary (ACTH), gonadal (estradiol, testosterone), and adrenal (cortisol) hormones.

Results: While adrenal and pituitary hormone levels were similar between groups at EF and OV, the MDD group showed a greater suppression of ACTH at OV compared to EF. Estradiol was greater in controls v. MDD at EF and OV. Controls showed greater activation compared to MDD in the hypothalamus, subgenual ACC, and OFC at both EF and OV, as well as the amygdala and hippocampus at OV.

Conclusions: These results suggest dysregulation of the HPA and HPG axes in MDD, with deficits in stress response regions involved in HPA-axis feedback mechanisms. With additional data on DHEA (produced by the adrenal cortex), future analyses will provide insights into where in the HPA axis these deficits occur.

1 Division of General Pediatrics, Boston Medical Center and Boston University School of Public Health 2 Department of Epidemiology, Tulane University School of Public Health and Tropical Medicine P-29 Childhood Adversities and Age at Menarche: Lessons From Post-War Britain

Background and Objective: Psychosocial stressors, including child abuse, paternal absence, and family conflict, are associated with age at menarche, yet pathways are poorly understood. Our objective is to investigate the impact of different forms of childhood adversities on age at menarche (≤12 and ≥14 years).

Methods: The 1958 British Birth Cohort is a prospective longitudinal cohort study (1958–2003). Using a factor analysis methodology, we identified six types of childhood adversities: financial, family dysfunction, lack of interest in education, lack of supportive caregiving, mental health/violence exposure, and family structure challenges. Logistic regression analyses to explore the impact of these six types of childhood adversities on age at menarche (≤12 and ≥14 years) among a subset of 4,524 female participants.

Results: Of the 4,524 female participants, 27% (n = 1,200) experienced menarche at ≥14 years. Cumulative adversities in childhood were associated with a graded increase in risk for later onset menarche (age ≥14) of 24% to 55% among those with 2 to ≥4 adversities. When evaluating individual adversity types, risk for menarche at ≥14 years increased by 19% to 74% for those exposed to financial hardships, lack of interest in education, and family violence/mental health issues. Family dysfunction was associated with decreased odds of menarche at ≥14 years of age (OR = 0.75, 95% CI 0.51–1.09, p = 0.13).

Conclusions: In this prospective cohort study, several categories of childhood adversities were associated with later menarcheal age. A graded association between cumulative adversities and risk for later age at menarche and protection from earlier menarche was demonstrated.

1 Department of Physical Therapy and Rehabilitation Science, University of California, San Francisco, Los Angeles 2 Department of Orthopedic Surgery and Department of Medicine, University of California, Los Angeles P-30 Hyperkyphosis Is Associated With Impaired Mobility Among Older Community-Dwelling Women: The Fracture Intervention Trial (FIT)

Background and Objective: Age-related hyperkyphosis is associated with impaired mobility, but it is not known what degree of hyperkyphosis predicts impairment. To determine the degree of hyperkyphosis associated with impaired mobility, we performed a cross-sectional analysis of 3,222 women ages 55–80 years who were randomized to placebo in the Fracture Intervention Trial (FIT), a randomized controlled trial of the effect of alendronate on risk for osteoporotic fractures.

Methods: Kyphosis angle was measured using the Debrunner kyphometer and was categorized into tertiles. Timed Up-and-Go test, a measure of functional mobility, was dichotomized into normal (=12 seconds) and impaired mobility (>12 seconds). Multivariate logistic regression models (controlling for age, smoking, spine bone mineral density, grip strength, and vertebral fractures) were used to determine cutpoints for kyphosis associated with impaired mobility.

Results: On average, women were 68 years old (SD = 6) with kyphosis angle of 48° (SD = 12°). Women in the third (worst) tertile of kyphosis (=54°) were 40% more likely to have impaired mobility (OR 1.4, 95% CI 1.1–2.0) than those in the first tertile were. Among the 1,048 women in the third tertile of kyphosis, 11% (355) displayed impaired mobility.

Conclusions: Kyphosis angle = 54° is associated with a 40% increased risk of impaired mobility. Hyperkyphosis is an easily identifiable clinical marker and may be a contributing factor to impaired mobility in the elderly.

1 Center for Healthy Aging, University of California, Davis, School of Medicine 2 Boston University School of Medicine 3 University of Alabama School of Medicine 4 University of Iowa School of Medicine 5 University of California, San Francisco, School of Medicine P-31 The Association of Parity with Knee Osteoarthritis and Total Knee Replacement in the MOST Cohort

Background and Objective: A recent study found that women's risk of total knee replacement (TKR) increased with each additional birth they had had. Our objective is to evaluate further the association of TKR and parity, and whether osteoarthritis (OA) may mediate this association.

Methods: The Multicenter Osteoarthritis Study (MOST), funded by the National Institutes of Health, is a longitudinal observational study of people ages 50 to 79 either with symptomatic knee OA or at high risk of knee OA. Women were grouped based on their number of children: 0, 1–3 births, and 4 or above. We used Poisson regression to estimate the relative risk of having baseline or incident radiographic knee OA (RKOA) and having prevalent TKR at 30 months. Generalized Estimating Equations were used to account for the correlation between two knees within a woman.

Results: A total of 1,820 women reported parity information (mean age 62.6 years, mean body mass index [BMI] 30.7). After adjusting for BMI and other covariates, compared to those who had no children, women with 1–3 children had a relative risk at 30 months of prevalent TKR of 2.4 (1.2, 4.8), while women with 4–12 children had a relative risk for prevalent TKR of 2.3 (1.1, 4.9). Parity was not significantly associated with baseline or incident RKOA.

Conclusions: Women's parity was associated with prevalent TKR but not with prevalent or incident RKOA. This suggests that the effect of parity on risk of TKR may result from social, neurological, or hormonal factors but not RKOA.

1 Rheumatology Division, The Feinberg School of Medicine, Northwestern University 2 Department of Internal Medicine, North Shore University Health System P-34 The Establishment of a Systemic Sclerosis Patient Registry: Lessons Learned

Background and Objective: Systemic sclerosis (SSc)/scleroderma is a rare disease that primarily affects women. The pathogenesis involves vasculopathy (Raynaud phenomenon), autoimmunity (autoantibody production), and fibrosis (internal organ dysfunction). SSc patients are an extraordinarily heterogeneous patient population demonstrating various clinical manifestations, rates of disease progression, patterns of internal organ involvement, response to treatment, and prognoses. Our objective is to define hitherto clinically indistinguishable subsets of SSc patients using clinical information as well as serum, DNA, and RNA assays for enrollment in SSc biomedical research projects.

Methods: We enroll consecutive SSc patients who fulfilled American College of Rheumatology criteria for diagnosis. Patients complete demographics forms and a detailed medication history, as well as health-related quality of life questionnaires. Patients may donate blood for serum collection and DNA isolation and tissue samples for RNA analyses. Standardized clinical assessments are completed by one of two scleroderma specialists. Evaluations include standardized laboratory and diagnostic testing (Doppler echocardiography, high-resolution computerized tomography of the chest, and pulmonary function testing). All information is stored in a state-of-the-art central data repository (EDW) that can be easily queried.

Results: DNA samples have been collected on 300/400 (75%) patients since March 2005: only 36 patients have declined. Since February 2009, 85 patients have enrolled in our registry and biorepository; only four patients have declined.

Conclusions: SSc patients seen at our tertiary care center are enrolling in our patient registry and biorepository. Physician and patient time constraints limit enrollment that necessitates collection/completion of separate components of the registry at separate clinic visits.

1 Department of Therapeutic Radiology, University of Minnesota Medical School 2 Department of Radiology, University of Minnesota Medical School 3 Department of Biostatistics, University of Minnesota Medical School 4 Department of Medicine, University of Minnesota Medical School 5 Department of Obstetrics and Gynecology, University of Minnesota Medical School P-35 Longitudinal Assessment of Bone Health in Gynecologic Oncology Patients: The Roles of Chemotherapy and Radiation

Background and Objective: Gynecologic cancer patients undergoing treatment combination of oophorectomy, chemotherapy, and radiation are at high risk for osteoporotic fracture leading to increased morbidities and reduced quality of life. Our objective is to evaluate the longitudinal changes of volumetric bone mineral density (vBMD) using quantitative computed tomography (QCT) in gynecologic cancer patients undergoing anticancer therapy.

Methods: Serial CT scans (pre- and post-treatment) were analyzed for vBMD in trabecular spine and the femoral neck. General linear mixed models were used to analyze the percentage vBMD loss for age, treatment, bone sites, scan interval, and their interactions.

Results: We evaluated 12 ovarian, 15 endometrial, and 18 cervical cancer patients. Mean age (±SD) was 65.1 (±11.6), 60.3 (±7.8), and 49.7 (±10.8), respectively. There was significant decrease in mean vBMD immediately after treatment (13.7% ± 20.3%, p < 0.0001), which continued to decrease for up to 1 year (12.0% ± 40.0%, p = 0.0003). Ovarian, endometrial, and cervical cancer patients lost mean vBMD (±SD) at the L1-L2 by 12.8% (±11.0%), −0.3% (±11.2%), and 14.2% (±9.9%) immediately after treatment; 15.9% (±19.6%), 5.2% (±20.5%), and 21.0% (±19.9%) after 1 year and at the femoral neck by 10.3% (±7.1%), 7.4% (±17.8%), and 10.2% (±13.8%) immediately after treatment; by 10.4% (±14.1%), 4.3% (±25.6%), and 3.6% (±9.5%), respectively, after 1 year. The mean vBMD decrease was greater at sites within the radiation treatment field than sites outside of the treatment field.

Conclusions: Significant bone loss in patients with gynecological cancers after chemotherapy or radiation treatment underlines the importance of monitoring treatment-induced bone loss and investigating preventive therapy.

1 Vanderbilt Epidemiology Center, Vanderbilt University School of Medicine 2 Shanghai Cancer Institute, Shanghai P-36 Resting Heart Rate and Risk of Type 2 Diabetes in Women

Background and Objective: Autonomic dysfunction may be involved in the pathogenesis of insulin resistance and the metabolic syndrome. Whether simple measures of autonomic activity, such as resting heart rate, may predict risk of type 2 diabetes (T2D) remains unclear. Our objective is to evaluate the association between resting heart rate and risk of T2D in the Shanghai Women's Health Study, a population-based prospective cohort study.

Methods: This analysis included 47,571 women without prior history of diabetes, cancer, cardiovascular disease, or thyroid dysfunction at the time when resting heart rate was measured. T2D was defined according to the American Diabetes Association criteria.

Results: During a mean followup of 4.9 years, 849 women developed T2D. Resting heart rate was positively and significantly associated with T2D risk. After adjustment for physical activity and other potential confounders, the hazard ratios (95% CIs) for T2D across increasing quintiles of heart rate were 1, 1.21 (0.99–1.47), 1.31 (1.05–1.62), 1.38 (1.12–1.70), and 1.61 (1.28–2.01), respectively. Further analyses of joint effects of heart rate with body mass index (BMI), waist-hip ratio (WHR), or blood pressure (BP) showed increased risk of T2D with increasing heart rate in all categories of BMI, WHR, or BP. Women in the highest category of heart rate plus highest category of BMI, WHR, or BP had the highest hazard ratios of T2D, ranging from 4.81 to 6.37.

Conclusions: A high resting heart rate is independently associated with an increased risk of T2D in women. The combination of heart rate with BMI, WHR, or BP improves T2D risk prediction.

1 Department of Sociology, Pennsylvania State University 2 The RAND Corporation, Pittsburgh P-38 Adolescent Obesity and Life Course Childbearing

Background and Objective: Obesity is negatively associated with a wide range of fertility outcomes among adult women, but research has not considered whether obesity has the same implications for adolescent women's childbearing experiences. Obese adolescent girls are less likely than non-obese adolescent girls to have sexual intercourse, but when they do have sex, obese adolescent girls are more likely than their non-obese counterparts to participate in high risk sexual behaviors that increase their risk of unintended pregnancy. Furthermore, fecundity is quite high in the teen years. The authors examined obese adolescent girls' risk of nonmarital pregnancy, timing of their first birth, and total number of children born by the end of their reproductive years. We hypothesize that adolescent obesity will be positively related to each childbearing outcome.

Methods: Twenty-five years of data from National Longitudinal Survey of Youth: 1979 (NLSY:79) on female respondents ages 16–19 in 1981 (n = 1,745) were analyzed to study hypotheses using descriptive statistics, logistic regression methods, and event history methods. Multivariate models adjusted for important confounders including poverty status and girls' race/ethnicity.

Results: Obese adolescent girls were found to have earlier first births and more total births than their non-obese counterparts in both descriptive and multivariate analyses.

Conclusions: Results suggest that scholars cannot assume that obesity has the same consequences for childbearing across the women's reproductive years. Instead, they must keep in mind that neither the consequences of weight nor women's reproductive capacity is static.

1 Pennsylvania State University College of Medicine 2 University of Pennsylvania School of Medicine P-39 TGF-β Dysregulation in Women with Polycystic Ovary Syndrome

Background and Objective: Polycystic Ovary Syndrome (PCOS) and TGF-β dysregulation have been individually associated with reproductive dysfunction, altered glucose metabolism, renin-angiotensin system (RAS) hyperactivity, and cardiovascular dysfunction. In addition, PCOS has been linked to Allele 8 (A8) of D19S884, a dinucleotide repeat in an intron of the fibrillin-3 gene. The fibrillins are extracellular matrix proteins that regulate TGF-β activity. Therefore, TGF-β dysregulation may play a central role in the pathogenesis of PCOS, particularly in the presence of A8. The objective of this study was to determine if circulating TGF-β levels are altered and associated with the presence of A8, RAS hyperactivity, and abnormal glucose metabolism in women with PCOS.

Methods: We measured circulating TGF-β levels and other hormonal and metabolic parameters in 120 age-matched women, including 40 PCOS women who possess A8 (A8+ PCOS), 40 PCOS women who do not possess A8 (A8- PCOS), and 40 normally menstruating women who do not have PCOS or A8 (A8- Controls).

Results: Compared with A8- Controls, A8- PCOS is associated with significantly higher TGF-β1 and lower TGF-β2, adjusting for BMI. Compared with A8- PCOS, A8+ PCOS is associated with significantly lower TGF-β1, similar TGF-β2, significantly higher aldosterone and Inhibin B, and a nonsignificant trend toward lower fasting glucose to insulin ratio, adjusting for BMI.

Conclusions: Among women with PCOS, A8 positivity may contribute to dysregulation of TGF-β1, increased RAS activity, altered glucose metabolism, and possibly reproductive dysfunction due to significantly higher Inhibin B levels. TGF-β2 levels are reduced in PCOS, regardless of A8 status.

1 Children's Hospital of Pittsburgh, University of Pittsburgh Medical Center 2 Western Psychiatric Institute and Clinic, University of Pittsburgh Medical Center P-42 Utilizing Ecological Momentary Assessment in Adolescents With Polycystic Ovary Syndrome To Quantify Behavior, Emotion, and Sleep

Background and Objective: This study examined the feasibility of using Ecological Momentary Assessment (EMA) to examine important domains relevant to interregulatory health processes in overweight adolescent females with Polycystic Ovary Syndrome (PCOS).

Methods: Participants were 20 overweight adolescent females engaged in a cognitive-behavioral and motivational interviewing intervention aimed at weight loss and improving mood (11–19 years old, 80% Caucasian, 15% African-American, mean BMI = 39). During this EMA protocol, participants were asked to report their physical activity (PA), nutrition, mood, and sleep during 14 cellular phone calls in their natural environment over three extended weekends (Thursday to Monday). Simultaneously, participants wore an actigraph (armband watch communicator) that provided instantaneous PA feedback (steps taken and kilocalories) and sleep parameters (duration and efficiency).

Results: EMA compliance rates for the armband and phone calls were 74.7% ± 0.3 and 64.2% ± 0.3, respectively. A positive association existed between baseline depression and armband compliance (Rho = 0.50, p = 0.03), but not phone call compliance (Rho = −0.26, p = 0.26). No relation was found between sleep problems and EMA compliance; a positive association existed between baseline measures of breathing regularity during sleep and armband compliance (Rho = 0.44, p = 0.052). No associations existed between baseline BMI and armband compliance (p = 0.99) or phone call compliance (p = 0.24).

Conclusions: Data from the armband and phone calls are presented to illustrate the depth of information acquired by utilizing this innovative methodology. These data reveal that nutrition, physical activity, mood, and sleep can be objectively measured simultaneously to enhance comprehensive treatment of overweight adolescents.

1 Department of Preventive Medicine and Public Health, University of Kansas Medical Center 2 Department of Dietetics and Nutrition, University of Kansas Medical Center 3 Schiefelbusch Institute for Life Span Studies, University of Kansas P-43 Phone-Based Weight Management for Rural Women

Background and Objective: Rural women have among the highest rates of obesity and sedentary lifestyle, yet few studies have examined strategies for delivering state-of-the-art obesity treatment to hard-to-reach rural areas. Phone-based treatment appears to be the best alternative to traditional face-to-face group meetings for rural populations of whom only half have home Internet access and where tele-video capacity is limited to a few sites. The purpose of this randomized study was to examine the impact and cost-effectiveness of a 6-month behavioral weight loss program delivered to rural women by phone, either one-on-one with a counselor or to a group via conference call.

Methods: Thirty-four rural women (mean BMI = 34.4, SD = 4.6) were randomized to group phone-based treatment or individual phone-based treatment.

Results: Completers analysis showed that weight loss was greater in the group condition (mean = 14.9 kgs, SD = 4.4) compared with the individual condition (mean = 9.5 kgs, SD = 5.2; p = 0.03). Among the total sample, 62% of participants in the group condition achieved the 10% weight loss goal compared to 50% in the individual condition. Group treatment was found to be more cost effective; using the percentage of participants who achieved 10% weight loss in intent-to-treat analysis, cost-effectiveness from a program plus participant perspective (average cost per successful participant was $714 for group counseling and $1029 for individual counseling).

Conclusions: Future research is warranted to examine the benefits of group phone-based treatment for long-term management of obesity among rural populations.

1 Oncology Division, University of Kansas Cancer Center 2 Department of Preventive Medicine, University of Kansas Medical Center 3 Department of Biostatistics, University of Kansas Medical Center P-44 Feasibility of a Diet, Exercise, and Behavior Modification Intervention for Post-Menopausal Breast Cancer Survivors

Background and Objective: Obesity is prevalent in general population and among breast cancer (BrCa) survivors. Weight at diagnosis and weight gain post-diagnosis are associated with increased risk of BrCa recurrence and mortality. Regular exercise also reduces the risk of dying from BrCa. This study was conducted to determine the feasibility of a structured diet/exercise/behavioral intervention to achieve a weight loss of >5%, and the effect of participation on serum biomarkers of BrCa and measures of overall health/fitness.

Methods: Subjects in this prospective pilot study are overweight (BMI > 25) female BrCa survivors. The 6-month combined modality diet/exercise/behavior modification intervention included 225 minutes per week of cardiovascular exercise plus resistance training; a 1,200–1,500 calories-per-day diet, including pre-packaged meals and low-calorie shakes; and a weekly in-person group behavioral meeting. Women were recruited into four cohorts of 10 to 15 (n = 50). Participants underwent pre- and post-intervention assessments: anthropometric measures, serum biomarkers, fitness test, and questionnaires to assess food frequency, fatigue, and quality of life.

Results: Women were successfully recruited and participated in the 6-month intervention. More than 95% of participants met the 5% weight loss goal at 6-months, and a majority of women reached an average weight loss of >10%. Mean age was 50, and time from diagnosis to on-study was about 45 months. A total of 85% of the women received adjuvant or neo-adjuvant chemotherapy, and 75% either were currently or had previously taken antihormonal therapy.

Conclusions: A 6-month diet/exercise/behavior modification intervention program is feasible and results in significant weight loss and percent body fat among overweight BrCa survivors.

1 Department of Family Medicine and Community Health, University of Minnesota Medical School 2 Department of Biostatistics, University of Minnesota 3 Department of Epidemiology and Community Health, University of Minnesota P-45 Parenting Style as a Predictor of Adolescent Weight and Weight-Related Behaviors: The Importance of the Opposite Sex Parent

Background and Objective: Current research indicates that specific parenting styles (authoritative, authoritarian, permissive neglectful) are associated with adolescent overweight, dietary intake, and physical activity, but the majority of research has been cross-sectional, making it difficult to determine the temporal order of these associations. Our objective is to (1) identify risk and protective factors of parenting style on adolescent weight status, dietary intake, and physical activity longitudinally; and (2) examine effects of gender differences between opposite-sex and same-sex parent-adolescent dyads.

Methods: Data from Project EAT, a population-based study with diverse ethnic and socioeconomic adolescents were used. Adolescents (N = 2,516) from 31 Minnesota schools completed in-class assessments in 1999 (Time 1) and mailed surveys in 2004 (Time 2). Multiple linear regression models were used to predict mean levels of adolescent outcomes at Time 2 from parenting style at Time 1.

Results: Maternal authoritative parenting style predicted lower BMI in sons and daughters compared to authoritarian and neglectful parenting styles. Paternal permissive parenting style predicted more fruits and vegetables intake in daughters compared to paternal authoritarian parenting style. We did not find significant associations between parenting style and adolescent physical activity.

Conclusions: Findings suggest that authoritative parenting style may play a protective role related to adolescent overweight and that the dimension of warmth and caring in the parent-adolescent relationship may be important in connection to adolescent healthy dietary intake. Further, several significant adolescent health outcomes were gender-specific to opposite-sex parent-adolescent dyads. This gender pairing may hold important information related to assessing and intervening with adolescent obesity.

1 Department of Pediatrics, University of Colorado, Denve 2 Division of Neurology, Oregon National Primate Research Center, Oregon Health and Sciences University P-46 Exposure to Maternal Obesity Does Not Reduce Mitochondrial Biogenesis or Complex Stoichiometry in Fetal Skeletal Muscle of Japanese Macaques

Background and Objective: Maternal obesity is associated with increased risk and earlier onset of obesity and type-2 diabetes (DM2) in the offspring, linking the fetal environment to adult susceptibility to metabolic diseases. Reduced mitochondrial density and impaired function have been observed in lean adult offspring of DM2 with insulin resistance, suggesting that these defects may begin during fetal development. Using a nonhuman primate model, we investigated whether intrauterine exposure to maternal obesity decreases fetal mitochondrial biogenesis and function in skeletal muscle.

Methods: Female Japanese macaques were fed either a control (10% fat) or a high-fat/calorie diet (35% fat; HFD) for 2–4 years. Skeletal muscle was collected from fetuses (F-CON/F-HFD) terminated early in the third-trimester from control and HFD mothers. Muscle gene expression was analyzed by DNA microarray and proteins involved in mitochondrial biogenesis and metabolic function confirmed by real-time PCR or immunoblot analysis.

Results: Skeletal muscle from F-HFD accumulated significantly more triglycerides with no difference in key regulators of lipid metabolism (ACC, ppar-a, ppar-γ). F-HFD muscle had increased expression of inflammatory/apoptotic pathways and reductions in MAPK-growth pathways, which was associated with smaller fetal weight (5–10%). However, no decrease was found in mitochondrial protein abundance, complex stoichiometry, or transcription factor expression (PGC1a, SIRT1/3) in F-HFD.

Conclusions: Intrauterine exposure to maternal obesity does not reduce skeletal muscle mitochondrial biogenesis or upregulate fat metabolism, despite increased intramuscular lipid and inflammation. Early fetal exposure to elevated lipids, prior to development of adipose depots, may later predispose offspring to impaired muscle fat utilization and insulin resistance.

1 Division of Rehabilitation Sciences, The University of Texas Medical Branch at Galveston 2 Sealy Center on Aging, The University of Texas Medical Branch at Galveston 3 Preventive Medicine and Community Health, The University of Texas Medical Branch at Galveston P-47 Gender Differences in Obesity and Life-Space Mobility among Older Mexican-Americans

Background and Objective: Obesity is a major public health problem associated with reduced mobility in older adults. Our objective is to examine gender differences between obesity (BMI = 30 Kg/m²) at baseline and life-space assessment (LSA) mobility at a 2-year followup among older Mexican-Americans.

Methods: A 2-year prospective study included Mexican-American men and women above age 75 and residing in five southwestern States. Data were collected on sociodemographic variables, physical function, medical conditions, BMI, health-related quality of life (HRQoL), and the LSA instrument that assessed the range, independency, and frequency of movement in the past 4 weeks preceding the interview in person.

Results: Out of 472 subjects, 30.6% of women and 16.7% of men had a BMI = 30 kg/m² at baseline. The mean score for the LSA instrument was 31.4 (SD = 20.3) in obese women and 40.7 (SD = 17.9) in obese men. Multiple regression analysis yielded a significant gender by obesity interaction effect (β = −8.34, p =0.03) on LSA mobility. Analysis stratified by gender showed the association between obesity and the LSA mobility in women (β = −5.96, p = 0.008) was partially mediated by physical function, HRQoL, and medical conditions. No association was found between obesity and LSA mobility in men.

Conclusions: The association between obesity and lower life-space mobility differed by sex in older Mexican-Americans. Among women, obesity maintained a negative and independent association with life-space mobility in models controlling for other factors that can influence mobility patterns. Conversely, obesity does not seem to affect mobility patterns of men.

1 Department of Pharmacy Practice, University of Illinois at Chicago 2 Center for Pharmaceutical Biotechnology, College of Pharmacy, University of Illinois at Chicago P-48 Hormonal Regulation of Hepatic Drug-Metabolizing Enzymes

Background and Objective: Medication use during pregnancy is prevalent. Pregnancy influences the rate and extent of hepatic drug metabolism, often leading to changes in dosage regimens. The underlying mechanisms remain unknown. Potentially, estrogen and progesterone, which increase in pregnancy, are responsible for the changes. Our objective is to determine the effects of estradiol and progesterone on expression of major drug-metabolizing enzymes in human hepatocytes.

Methods: Human hepatocytes were treated with ethanol, estradiol, or progesterone for 60 hours. mRNA expression levels of cytochrome P450 (CYP) enzymes (CYP1A2, −2A6, −2B6, −2C9, −2C19, −2D6, −2E1, and −3A4) were determined by quantitative real-time polymerase chain reaction. After estradiol treatment, activities of CYP2B6, −2C9, −2C19, −2D6, and −3A4 were determined in human hepatocytes by performing functionality assays using a set of probe drugs.

Results: Estradiol treatment of human hepatocytes significantly decreased mRNA levels of CYP1A2 and increased mRNA expression of CYP2A6 and −2B6. The estradiol-mediated changes in mRNA expression of other CYP enzymes were not significant. Activities of CYP2B6, −2C9, and −3A4 were significantly increased by estradiol, whereas activities of other CYP enzymes were not affected. Progesterone significantly increased mRNA expression of CYP2A6, −2B6, and −3A4.

Conclusions: The pattern of hormone-mediated changes in expression or activity of most CYPs in human hepatocytes was similar to changes occurring during pregnancy. This suggests that estrogen and progesterone may be responsible for altered drug metabolism in pregnancy. These findings should help us determine optimal drug dosage regimens through a better understanding of how drug metabolism is altered in pregnancy.

1 Department of Medical Microbiology and Immunology, University of California, Davis 2 Department of Obstetrics and Gynecology, University of California, Davis 3 Kaiser Permanente Medical Group, Sacramento 4 Internal Medicine, University of California, Davis P-50 Effects of Aging and Menopause on Gut-Associated Lymphoid Tissue

Background and Objective: About 15.4 million women are living with Human Immunodeficiency Virus (HIV) infection worldwide. HIV infection results in the loss of CD4+ T cells in the peripheral blood and gut-associated lymphoid tissue (GALT). HIV disease progression correlates with immune activation. A growing proportion of HIV-infected women are now over 50 years old. The effects of aging and menopause on GALT are unknown. With chronic inflammation, such as HIV infection, the pathogenic effects of HIV may be exacerbated during the course of aging and menopause, as low-grade systemic inflammation is associated with both of these conditions. Our objective is to compare the effects of gender and menopause on cellular activation and inflammation in GALT of HIV negative individuals.

Methods: We evaluated peripheral blood and mucosal T lymphocyte subsets in HIV negative participants using Flow Cytometry. Gene expression profiles in GALT in male (21–57 years) and female (23–59 years) volunteers were analyzed using microarray technology, real-time PCR, and Robust Multi-Chip-based statistical criteria.

Results: Postmenopausal women (n = 3) had higher percentages (18–48% v. 2.5–5%) of activated CD4+ and CD8+ T cells in blood and GALT compared to premenopausal women (n = 3) and men (n = 6). We observed greater than a twofold increase in gene expression associated with inflammation (IL6, iNOS2A, Catalase) and T cell activation (NFAT3, STAT3, IL2, IL12) in postmenopausal women compared to premenopausal women and age-matched men.

Conclusions: Menopause in women may be associated with increased immune activation, providing the foundation for investigation of the independent effects of menopause on HIV pathogenesis.

1 Department of Medicinal Chemistry and Pharmacognosy, University of Illinois at Chicago 2 University of Illinois at Chicago/National Institutes of Health Center for Botanical Dietary Supplements Research, Department of Medicinal Chemistry and Pharmacognosy, University of Illinois at Chicago P-51 Evaluation of Phytoprogestins in Botanical Extracts

Background and Objective: Menopausal women are consuming botanical supplements for the alleviation of hot flashes. Estrogen-like compounds in botanicals used to treat menopausal symptoms might increase the risk for developing endometrial cancer similar to standard estrogens. Progesterone is known to prevent estrogen-induced uterine cancer and is typically combined with estrogen in hormone replacement therapies. Our objective is to determine if plant extracts commonly used to treat menopausal symptoms contain novel progestins that might be used in combination with plant-based estrogens.

Methods: Eight botanical extracts (75% ethanolic) were tested for their ability to induce a luciferase reporter gene driven by the progesterone responsive element promoter. Botanical extracts were then investigated to determine if they could displace fluorescently labeled progesterone from the receptor-binding pocket of the progesterone receptor. Progesterone-regulated gene induction from endogenous promoters was investigated with quantitative real-time PCR.

Results: A 75%-ethanolic extract of red clover bound to the progesterone receptor and significantly induced a luciferase reporter gene linked to the progesterone responsive element in breast cancer cells. The red clover extract also significantly induced endogenous genes regulated by progestins measured using real-time PCR. A library of compounds previously identified in red clover was screened, and two flavonoids, kaempferol and naringenin, were found with a high affinity for the progesterone receptor. Future work will confirm if kaempferol and naringenin have progesterone-like activity in animal models.

Conclusions: Red clover contains phytoprogestins that might protect against endometrial cancers or provide a safer progestin for applications in women's health.

1 Division of Endocrinology, Diabetes, and Hypertension, Brigham and Women's Hospital, Harvard Medical School 2 Harvard Reproductive Endocrine Sciences Center 3 Division of Endocrinology, Children's Hospital Boston P-53 Critical Requirements of Reproductive Hormones for Proper Organization and Activation of the Hypothalamic Kisspeptin System: Evidence From GnRH-Deficient hpg Mice

Background and Objective: Women-specific health concerns indicate that critical periods during sexual differentiation can affect lifelong fitness. The neuropeptide kisspeptin potently stimulates GnRH release, influencing sexual differentiation, puberty, and fertility. In GnRH-deficient hpg mice, kisspeptin immunostaining in the hypothalamic nuclei, AVPV and ARC, suggested both reproductive hormone-dependent and hormone-independent contributions to pubertal kisspeptin expression. To further understand developmental influences on kisspeptin neurons, hpg mice were used to compare changes in kisspeptin immunostaining to changes in Kiss1 gene expression, and to test the hypothesis that exogenous estradiol affects hpg AVPV and ARC kisspeptin similarly to wildtype.

Methods: Kiss1 expression and kisspeptin immunostaining were compared in hpg and wildtype mice during maturation. To test response to exogenous estradiol, ovariectomized hpg and wildtype females were treated with estradiol for 1 week and, then, immunostained for kisspeptin.

Results: Kiss1 expression in the AVPV corroborated immunostaining results, demonstrating a loss of sexual-dimorphism in hpg mice. hpg ARC Kiss1 mRNA was elevated at pubertal ages, emphasizing the lack of sex steroid negative feedback. AVPV kisspeptin increased in ovariectomized wildtype females in response to estradiol treatment in a manner consistent with a positive-feedback response; estradiol-treated hpg females displayed no differences. Alternatively, estradiol treatment restored normal ARC kisspeptin staining patterns in both ovariectomized wildtype and hpg females, demonstrating that negative-feedback responses to sex steroids were intact.

Conclusions: Activation of the kisspeptinergic system is dependent upon developmental exposure to reproductive hormones. The blunted positive feedback in hpg females implies gonadal hormones are required to shape the neural mechanism necessary for ovulation.

1 Division of Cardiovascular Medicine, Oregon Health & Science University ² Department of Pediatrics, University of Iowa Carver College of Medicine P-54 Cardiac Consequences of Transfusion Following Fetal Anemia During Ovine Pregnancy

Background and Objective: Fetal anemia is a serious complication of pregnancy that can alter cardiac growth and cardiovascular physiology, and lead to fetal demise. Transfusion can improve fetal health and maternal reproductive outcome, but the cardiac consequences of recovery from anemia are unknown. For fetal hearts enlarged by anemia, we predict their mass would become normalized within 10 days following transfusion. We hypothesized that accelerated cardiac growth of anemic fetuses would cease following transfusion, and that heart weights would consequently normalize.

Methods: Fetal sheep were made anemic by isovolumetric hemorrhage starting at 75% gestation. After 10 days, fetuses were either euthanized (n = 6) or transfused to restore a normal hematocrit for 10 days, and then euthanized (n = 7). Age-matched control fetuses were studied for both experimental groups (n = 5 and n = 7, respectively). Morphometric measurements were made of isolated cardiomyocytes.

Results: Hearts (adjusted for body weight) were 31% heavier in anemic v. age-matched fetuses. Adjusted heart weight 10 days after fetal transfusion remained 25% heavier than age-matched controls. Cardiomyocyte sizes, cell cycle activity, and terminal differentiation were not different between experimental groups and their controls, and do not fully explain the sustained cardiac hypertrophy.

Conclusions: Ten days of anemia at 75% gestation causes cardiac enlargement by cardiomyocyte proliferation. Transfusion of anemic fetuses did not lead to normalization of adjusted heart weight in the recovery period. We conclude that the fetal heart responds rapidly to the adverse stimulus of anemia, but does not respond rapidly to correction of this cardiovascular stress.

1 Department of Pediatrics, University of California, Davis, School of Medicine 2 Harvard School of Public Health 3 Family Violence Prevention Fund P-56 Pregnancy Control and Partner Violence among Adolescent Female Users of Family Planning Clinics

Background and Objective: Intimate partner violence (IPV—physical or sexual violence by a partner) has been associated with adolescent and unintended pregnancy. One mechanism that may explain how IPV relates to poor reproductive health is male partners' control of women's reproduction through condom refusal, abusive responses to contraceptive negotiation, pressuring women to get pregnant, keeping women from abortion, and birth control sabotage; cumulatively, these behaviors constitute pregnancy control. The extent of pregnancy control experienced by adolescents is not known. Our objective is to determine the prevalence of pregnancy control among adolescent females using confidential clinics and the association of pregnancy control with IPV.

Methods: Female clients ages 16 to 20 years (N = 567) were recruited from five family planning clinics in California to complete a 20-minute audio computer-assisted self-interview in English or Spanish about reproductive control, IPV, and pregnancy experiences.

Results: The lifetime prevalence of physical or sexual violence from an intimate partner was 50.1%. Pregnancy control was reported by 28.9% of participants (including 21.5% of clients who reported no history of IPV). IPV was associated with experiences of pressuring to prevent abortion (OR 2.25, 95% CI 1.40–3.61), pregnancy control (OR 2.08, 95% CI 1.43–3.03), and birth control sabotage specifically (OR 2.34, 95% CI 1.46–3.75). However, IPV was not associated with unintended pregnancy (OR 1.28, 95% CI 0.88–1.87).

Conclusions: IPV and pregnancy control are common among adolescent female clients of family planning clinics and are associated with each other. The impact of partners' pregnancy control on adolescent reproductive and sexual health outcomes merits further study.

1 Department of Obstetrics, Gynecology and Reproductive Sciences 2 Department of Epidemiology, University of Pittsburgh 3 Department of Epidemiology, Institute of Public Health, University of Aarhus 4 Department of Epidemiology, University of California, Los Angeles P-58 Severity and Recurrence of Preterm Birth and Maternal Cardiovascular Disease Risk

Background and Objective: Women with preterm births (PTB) may have excess cardiovascular disease (CVD) risk, but mechanisms linking these conditions are unknown. We hypothesized that a common predisposition to both conditions would be reflected by a more pronounced relationship between early or recurrent PTB and CVD, as these tend to aggregate in families.

Methods: We related PTB severity (earlier gestational age at delivery) and recurrence (at least two births) among women who gave birth between 1973 and 1983 in Denmark (n = 426,865) to maternal CVD morbidity and mortality. Birth data were linked to CVD hospitalizations and deaths identified in national registers via ICD codes (1977 to 2006) and data were analyzed using Cox proportional hazards models.

Results: Women with a prior PTB had excess CVD after adjustment for age, parity, education, and birth year (HR 1.26, 95% CI 1.22–1.31). This was only modestly attenuated when women with preeclampsia or small for gestational age births were excluded, and the relationship was stronger for CVD mortality (HR 1.91 [1.67, 2.18]). Among women with only one birth, CVD risk increased with early (<32 weeks, HR 1.66 [1.44, 1.91]) v. moderate (35–36 weeks, HR 1.39 [1.30, 1.48]) preterm births. Among women with at least two births, recurrent PTB was associated with modestly higher risk for CVD compared to one PTB. Sensitivity analysis suggested that confounding by smoking only partly explained these associations.

Conclusions: Associations between PTB and later life maternal CVD could reflect a common underlying predisposition related to both conditions. Preterm births may identify women at increased risk for CVD.

1 Department of Biochemistry and Biophysics, University of North Carolina School of Medicine 2 Division of Hematology and Oncology of the Department of Medicine, University of North Carolina School of Medicine 3 Placental Analytics 4 Department of Obstetrics and Gynecology, University of North Carolina School of Medicine P-59 Placental Pathology and Pregnancy Outcome in Sickle Cell Disease: A Pilot Study

Background and Objective: Pregnancy complications in women with sickle cell disease (SCD) include preterm birth, delivery of small for gestational age (SGA) infants, and potential hypertension of pregnancy. The role that placental pathology may play in these complications has never been examined. We sought to explore a potential connection between placental inflammatory and vascular pathology and pregnancy outcome in SCD.

Methods: For this pilot study, we obtained pregnancy outcome, blood pressure, and placental pathology data from the Pregnancy, Infection and Nutrition study, from which 23 subjects were identified as having SCD and placental pathology data were available for six of them. Placental pathology was divided into vascular and inflammatory categories, and a severity score was assigned to pathology as 0 (none) to 4 (pronounced).

Results: Three out of the six placentas from pregnant women with SCD had some degree of pathology, though none was pronounced. Of those SCD placentas with no pathology detected, there was only one instance of delivery of an SGA infant, no preterm births, and no rise in systolic blood pressure across pregnancy. The median rise in systolic blood pressure in subjects with placental pathology was 15.0 mmHg (range, 8, 24). Both cases of preterm birth occurred in subjects with either mild vascular and inflammatory placental pathology or moderate vascular pathology. The subject with both vascular and inflammatory placental pathology demonstrated the worst pregnancy outcome.

Conclusions: Placental pathology, especially multiple comorbid types of placental pathology, may play a role in poor pregnancy outcome in SCD.

1 Department of Obstetrics, Gynecology, and Reproductive Sciences 2 Department of Biochemistry and Molecular Biology, University of Maryland School of Medicine P-60 Thioredoxin Ameliorates Hyperglycemia-Induced Embryonic Vasculopathy via Blockade of the Pathways Leading to Apoptosis

Background and Objective: Congenital malformations occur in up to 10% of babies born to diabetic women. Hyperglycemia causes embryonic vasculopathy that leads to embryonic lethality or malformations. Maternal diabetes induces apoptosis in Flk1+ progenitors and reduces the number of blood islands resulting in aberrant vasculogenesis. Maternal diabetes induces embryonic vasculopathy through two distinct sets of molecular events: activation of the ASK1/iNOS pathway and reduction of HIF-1α/VEGF pathway. Thioredoxin (Trx) is an antiapoptotic and antioxidative protein. Diabetic embryonic vasculopathy is an oxidative stress and apoptotic disease process. Therefore, we reason that Trx may have preventive effects on this disease process. Our objective is to examine the effect of Trx on hyperglycemia-induced aberrant pathways and consequent vasculopathy.

Methods: Embryonic day 7 mouse conceptuses were cultured under euglycemic (150 mg/dl) and hyperglycemic (300 mg/dl glucose) conditions with or without 0.1, 0.5, or 2.0 μg/ml Trx. Yolk sac vasculatures of 48-hour-cultured conceptuses were morphologically assessed. Levels of pASK1 and HIF-1α were determined in 24-hour-cultured conceptuses with or without 2 μ g Trx treatment.

Results: Hyperglycemia significantly decreased vasculature morphological scores. Vasculature morphological scores in conceptuses cultured in hyperglycemia plus 0.5 or 2 μg/ml Trx were significantly higher than those in conceptuses cultured in hyperglycemia alone. Hyperglycemia reduced HIF-1α levels, whereas Trx treatment reversed hyperglycemia-reduced HIF-1α levels. Hyperglycemia significantly increased the levels of pASK1, and Trx blocked hyperglycemia-induced ASK1 activation.

Conclusions: Trx can block ASK1 activation and also reverse HIF-1α reduction induced by hyperglycemia, thereby preventing apoptosis and subsequent embryonic vasculopathy.

1 Perinatal Research Center, University of Colorado, Denver 2 Department of Pediatrics, University of Colorado, Denver P-62 Intrauterine Growth Restriction Results in Increased Insulin-Stimulated Proliferation but Not Hypertrophy in Fetal Myocytes

Background and Objective: Reduced fetal skeletal muscle growth is a hallmark of intrauterine growth restriction (IUGR) and leads to increased risk for adult insulin resistance; however, the causative mechanisms are unknown. Our objective is to determine the effect of insulin on cellular proliferation and protein synthesis in primary cultured myocytes from ovine IUGR and control fetuses.

Methods: Studies were performed using an established ovine model of placental insufficiency and IUGR. Fetal myoblasts were harvested from control (n = 6) and IUGR (n = 3) fetuses at ∼90% of gestation, cultured in increasing insulin concentrations, and counted at 3 and 5 days to measure cell proliferation. Fully differentiated myotubes were serum starved overnight and incubated in insulin for up to 6 hours to measure protein synthesis rates using L-[3,4,5-3H(N)] leucine incorporation.

Results: IUGR fetuses weighed 43% less, had 67% percent lower blood oxygen contents, and had 40% lower plasma glucose concentrations compared to control fetuses (p < 0.01). Insulin increased proliferation in cells from both control and IUGR fetuses; however, the proliferation rate was twofold higher in IUGR cells (p = 0.005). Insulin increased protein synthesis rates in a dose-dependant manner (p = 0.009), but the responses did not differ between control and IUGR.

Conclusions: Myoblasts from IUGR fetuses demonstrate strikingly increased capacity for proliferation in response to insulin compared to cells from control fetuses; however, this is not associated with a further increase in protein synthesis in mature myotubes. Thus, IUGR fetal myocytes have a strong capacity for growth (myogenesis) when exposed to insulin, but limited ability for catch-up growth by hypertrophy.