Structural differences contributing to sex-specific associations between FN BMD and whole-bone strength for adult White women and men.

Hip areal BMD (aBMD) is widely used to identify individuals with increased fracture risk. Low aBMD indicates low strength, but this association differs by sex with men showing greater strength for a given aBMD than women. To better understand the structural basis giving rise to this sex-specific discrepancy, cadaveric proximal femurs from White female and male donors were imaged using nano-CT and loaded in a sideways fall configuration to assess strength. FN pseudoDXA images were generated to identify associations among structure, aBMD, and strength that differ by sex. Strength correlated significantly with pseudoDXA aBMD for females (R2 = 0.468, P < .001) and males (R2 = 0.393, P < .001), but the elevations (y-intercepts) of the linear regressions differed between sexes (P < .001). Male proximal femurs were 1045 N stronger than females for a given pseudoDXA aBMD. However, strength correlated with pseudoDXA BMC for females (R2 = 0.433, P < .001) and males (R2 = 0.443, P < .001) but without significant slope (P = .431) or elevation (P = .058) differences. Dividing pseudoDXA BMC by FN-width, total cross-sectional area, or FN-volume led to significantly different associations between strength and the size-adjusted BMC measures for women and men. Three structural differences were identified that differentially affected aBMD and strength for women and men: First, men had more bone mass per unit volume than women; second, different cross-sectional shapes resulted in larger proportions of bone mass orthogonal to the DXA image for men than women; and third, men and women had different proportions of cortical and trabecular bone relative to BMC. Thus, the proximal femurs of women were not smaller versions of men but were constructed in fundamentally different manners. Dividing BMC by a bone size measure was responsible for the sex-specific associations between hip aBMD and strength. Thus, a new approach for adjusting measures of bone mass for bone size and stature is warranted.

Bone quality following peripubertal growth in a mouse model of transmasculine gender-affirming hormone therapy.

During peri-puberty, bone growth and the attainment peak bone mass is driven predominantly by sex steroids. This is important when treating transgender and gender diverse youth, who have become increasingly present at pediatric clinics. Analogues of gonadotropin-releasing hormone (GnRH) are commonly prescribed to transgender and gender diverse youth prior to starting gender-affirming hormone therapy (GAHT). However, the impact of GnRH agonists on long bones with the addition of GAHT is relatively unknown. To explore this, we developed a trans-masculine model by introducing either GnRHa or vehicle treatment to female-born mice at a pre-pubertal age. This treatment was followed by male GAHT (testosterone, T) or control treatment three weeks later. Six weeks after T therapy, bone quality was compared between four treatment groups: Control (vehicle only), GnRHa-only, GnRHa + T, and T-only. Bone length/size, bone shape, mechanical properties, and trabecular morphology were modulated by GAHT. Independent of GnRHa administration, mice treated with T had shorter femurs, larger trabecular volume and increased trabecular number, higher trabecular bone mineral density, and wider superstructures on the surface of bone (e.g., third trochanters) when compared to control or GnRHa-only mice. In conclusion, prolonged treatment of GnRHa with subsequent GAHT treatment directly affect the composition, parameters, and morphology of the developing long bone. These findings provide insight to help guide clinical approaches to care for transgender and gender diverse youth.

Optogenetic-induced muscle loading leads to mechanical adaptation of the Achilles tendon enthesis in mice.

Skeletal shape depends on the transmission of contractile muscle forces from tendon to bone across the enthesis. Loss of muscle loading impairs enthesis development, yet little is known if and how the postnatal enthesis adapts to increased loading. Here, we studied adaptations in enthesis structure and function in response to increased loading, using optogenetically induced muscle contraction in young (i.e., growth) and adult (i.e., mature) mice. Daily bouts of unilateral optogenetic loading in young mice led to radial calcaneal expansion and warping. This also led to a weaker enthesis with increased collagen damage in young tendon and enthisis, with little change in adult mice. We then used RNA sequencing to identify the pathways associated with increased mechanical loading during growth. In tendon, we found enrichment of glycolysis, focal adhesion, and cell-matrix interactions. In bone, we found enrichment of inflammation and cell cycle. Together, we demonstrate the utility of optogenetic-induced muscle contraction to elicit in vivo adaptation of the enthesis.

Associations Among Hip Structure, Bone Mineral Density, and Strength Vary With External Bone Size in White Women.

Bone mineral density (BMD) is heavily relied upon to reflect structural changes affecting hip strength and fracture risk. Strong correlations between BMD and strength are needed to provide confidence that structural changes are reflected in BMD and, in turn, strength. This study investigated how variation in bone structure gives rise to variation in BMD and strength and tested whether these associations differ with external bone size. Cadaveric proximal femurs (n = 30, White women, 36-89+ years) were imaged using nanocomputed tomography (nano-CT) and loaded in a sideways fall configuration to assess bone strength and brittleness. Bone voxels within the nano-CT images were projected onto a plane to create pseudo dual-energy X-ray absorptiometry (pseudo-DXA) images consistent with a clinical DXA scan. A validation study using 19 samples confirmed pseudo-DXA measures correlated significantly with those measured from a commercially available DXA system, including bone mineral content (BMC) (R 2  = 0.95), area (R 2  = 0.58), and BMD (R 2  = 0.92). BMD-strength associations were conducted using multivariate linear regression analyses with the samples divided into narrow and wide groups by pseudo-DXA area. Nearly 80% of the variation in strength was explained by age, body weight, and pseudo-DXA BMD for the narrow subgroup. Including additional structural or density distribution information in regression models only modestly improved the correlations. In contrast, age, body weight, and pseudo-DXA BMD explained only half of the variation in strength for the wide subgroup. Including bone density distribution or structural details did not improve the correlations, but including post-yield deflection (PYD), a measure of bone material brittleness, did increase the coefficient of determination to more than 70% for the wide subgroup. This outcome suggested material level effects play an important role in the strength of wide femoral necks. Thus, the associations among structure, BMD, and strength differed with external bone size, providing evidence that structure-function relationships may be improved by judiciously sorting study cohorts into subgroups. © 2022 The Authors. JBMR Plus published by Wiley Periodicals LLC on behalf of American Society for Bone and Mineral Research.

Bone Morphology and Vascular Supply of Pedicled Distal Radius Bone Using Nano-Computed Tomography.

BACKGROUND: The goal of this study was to use nano-computed tomography to describe the intraosseous vascularity and structural characteristics of commonly used distal radius vascularized bone grafts for treatment of scaphoid nonunion. METHODS: We obtained 8 fresh frozen human cadaver forearm specimens for infusion of barium contrast. Specimens were scanned and segmented to quantify the vascular volume and trabecular density within 3 common graft regions, including 1, 2 intercompartmental supraretinacular artery (1,2 ICSRA), fourth extensor compartment artery (4 ECA), and volar carpal artery (VCA), as well as thirds of the scaphoid. Outcomes also included mean and maximum cortical thickness and number of cortical perforators. Single-specimen analyses were also performed comparing vascularity and trabecular density of each graft with scaphoid regions of a single specimen. Statistical analysis was performed using analysis of variance with post hoc Tukey testing when P value was less than .05. RESULTS: There was no significant difference between groups in the mean percent vascularity (P = .76). The ratio of trabecular bone in each graft to scaphoid thirds was less than 1. The mean cortical thickness (0.79 mm, 95% confidence interval [CI], 0.66-0.93 mm) and maximum cortical thickness (1.45 mm, 95% CI, 1.27-1.63 mm) of VCA grafts were both significantly greater than those of 4 ECA and 1,2 ICSRA (P < .001). CONCLUSIONS: There were no differences between vascular density of the 3 grafts and the scaphoid. Pedicled distal radius bone grafts have similar vascularity but morphometric differences such as cortical thickness and trabecular density which have unclear clinical implications.

Osteochondral Allografts for Large Oval Defects of the Medial Femoral Condyle: A Comparison of Single Lateral Versus Medial Femoral Condyle Oval Grafts Versus 2 Overlapping Circular Grafts.

BACKGROUND: Studies have demonstrated the acceptability of using a contralateral nonorthotopic lateral femoral condyle (LFC) graft for a circular medial femoral condyle (MFC) osteochondral defect up to 20 to 25 mm in diameter. Larger oblong defects can now be managed using either overlapping circle grafts or a single oblong-shaped osteochondral allograft (OCA). PURPOSE: To determine if an oblong contralateral nonorthotopic LFC OCA can attain an acceptable surface contour match compared with an oblong ipsilateral MFC OCA or an overlapping circle technique for large oblong defects of the MFC. STUDY DESIGN: Controlled laboratory study. METHODS: A total of 120 fresh-frozen human femoral condyles were matched by tibial width into 30 groups of 4 condyles (1 recipient MFC, 3 donor condyles). The recipient MFC was initially imaged using nano-computed tomography (nano-CT). A 17 × 36-mm oblong defect was created in the recipient MFC. Overall, 3 donor groups were formed: MFC oblong, LFC nonorthotopic oblong, LFC or MFC overlapping circles. After each transplant, the recipient condyle underwent nano-CT and was digitally reconstructed, which was superimposed on the initial nano-CT scan of the native recipient condyle. Dragonfly 3D software was used to determine the root mean square (RMS) of both the surface height deviation and the circumferential step-off height deviation between the native and donor cartilage surfaces for each graft. RESULTS: RMS surface height deviations were as follows: 0.59 mm for MFC oblong grafts, 0.58 mm for LFC oblong grafts, and 0.78 mm for overlapping circle grafts. The MFC and LFC oblong grafts had significantly less surface height deviation than the overlapping circle grafts (P = .004 and P = .002, respectively). RMS step-off height deviations were as follows: 0.68 mm for MFC oblong grafts, 0.70 mm for LFC oblong grafts, and 0.85 mm for overlapping circle grafts. The MFC and LFC oblong grafts had significantly less step-off height deviation than the overlapping circle grafts (P < .001 and P = .002, respectively). The majority of this difference was on the medial segment of the overlapping circle grafts. CONCLUSION: Oblong ipsilateral MFC OCAs and oblong contralateral nonorthotopic LFC OCAs produced a significantly better surface contour match to the native MFC than overlapping circle grafts for oblong defects 17 × 36 mm in size. CLINICAL RELEVANCE: Size-matched contralateral nonorthotopic LFC grafts are acceptable for MFC defects, which may allow for a quicker match, earlier patient care, and less wastage of valuable donor tissue.

Sex and External Size Specific Limitations in Assessing Bone Health From Adult Hand Radiographs.

Morphological parameters measured for the second metacarpal from hand radiographs are used clinically for assessing bone health during growth and aging. Understanding how these morphological parameters relate to metacarpal strength and strength at other anatomical sites is critical for providing informed decision-making regarding treatment strategies and effectiveness. The goals of this study were to evaluate the extent to which 11 morphological parameters, nine of which were measured from hand radiographs, relate to experimentally measured whole-bone strength assessed at multiple anatomical sites and to test whether these associations differed between men and women. Bone morphology and strength were assessed for the second and third metacarpals, radial diaphysis, femoral diaphysis, and proximal femur for 28 white male donors (18-89 years old) and 35 white female donors (36-89+ years old). The only morphological parameter to show a significant correlation with strength without a sex-specific effect was cortical area. Dimensionless morphological parameters derived from hand radiographs correlated significantly with strength for females, but few did for males. Males and females showed a significant association between the circularity of the metacarpal cross-section and the outer width measured in the mediolateral direction. This cross-sectional shape variation contributed to systematic bias in estimating strength using cortical area and assuming a circular cross-section. This was confirmed by the observation that use of elliptical formulas reduced the systematic bias associated with using circular approximations for morphology. Thus, cortical area was the best predictor of strength without a sex-specific difference in the correlation but was not without limitations owing to out-of-plane shape variations. The dependence of cross-sectional shape on the outer bone width measured from a hand radiograph may provide a way to further improve bone health assessments and informed decision making for optimizing strength-building and fracture-prevention treatment strategies. © 2022 The Authors. JBMR Plus published by Wiley Periodicals LLC on behalf of American Society for Bone and Mineral Research.

In vivo quantitative imaging biomarkers of bone quality and mineral density using multi-band-SWIFT magnetic resonance imaging.

Bone is a composite biomaterial of mineral crystals, organic matrix, and water. Each contributes to bone quality and strength and may change independently, or together, with disease progression and treatment. Even so, there is a near ubiquitous reliance on ionizing x-ray-based approaches to measure bone mineral density (BMD) which is unable to fully characterize bone strength and may not adequately predict fracture risk. Characterization of treatment efficacy in bone diseases of altered remodeling is complicated by the lack of imaging modality able to safely monitor material-level and biochemical changes in vivo. To improve upon the current state of bone imaging, we tested the efficacy of Multi Band SWeep Imaging with Fourier Transformation (MB-SWIFT) magnetic resonance imaging (MRI) as a readout of bone derangement in an estrogen deficient ovariectomized (OVX) rat model during growth. MB-SWIFT MRI-derived BMD correlated significantly with BMD measured using micro-computed tomography (µCT). In this rodent model, growth appeared to overcome estrogen deficiency as bone mass continued to increase longitudinally over the duration of the study. Nonetheless, after 10 weeks of intervention, MB-SWIFT detected significant changes consistent with estrogen deficiency in cortical water, cortical matrix organization (T1), and marrow fat. Findings point to MB-SWIFT's ability to quantify BMD in good agreement with µCT while providing additive quantitative outcomes about bone quality in a manner consistent with estrogen deficiency. These results indicate MB-SWIFT as a non-ionizing imaging strategy with value for bone imaging and may be a promising technique to progress to the clinic for monitoring and clinical management of patients with bone diseases such as osteoporosis.

External Bone Size Is a Key Determinant of Strength-Decline Trajectories of Aging Male Radii.

Given prior work showing associations between remodeling and external bone size, we tested the hypothesis that wide bones would show a greater negative correlation between whole-bone strength and age compared with narrow bones. Cadaveric male radii (n = 37 pairs, 18 to 89 years old) were evaluated biomechanically, and samples were sorted into narrow and wide subgroups using height-adjusted robustness (total area/bone length). Strength was 54% greater (p < 0.0001) in wide compared with narrow radii for young adults (<40 years old). However, the greater strength of young-adult wide radii was not observed for older wide radii, as the wide (R2 = 0.565, p = 0.001), but not narrow (R2 = 0.0004, p = 0.944) subgroup showed a significant negative correlation between strength and age. Significant positive correlations between age and robustness (R2 = 0.269, p = 0.048), cortical area (Ct.Ar; R2 = 0.356, p = 0.019), and the mineral/matrix ratio (MMR; R2 = 0.293, p = 0.037) were observed for narrow, but not wide radii (robustness: R2 = 0.015, p = 0.217; Ct.Ar: R2 = 0.095, p = 0.245; MMR: R2 = 0.086, p = 0.271). Porosity increased with age for the narrow (R2 = 0.556, p = 0.001) and wide (R2 = 0.321, p = 0.022) subgroups. The wide subgroup (p < 0.0001) showed a significantly greater elevation of a new measure called the Cortical Pore Score, which quantifies the cumulative effect of pore size and location, indicating that porosity had a more deleterious effect on strength for wide compared with narrow radii. Thus, the divergent strength-age regressions implied that narrow radii maintained a low strength with aging by increasing external size and mineral content to mechanically offset increases in porosity. In contrast, the significant negative strength-age correlation for wide radii implied that the deleterious effect of greater porosity further from the centroid was not offset by changes in outer bone size or mineral content. Thus, the low strength of elderly male radii arose through different biomechanical mechanisms. Consideration of different strength-age regressions (trajectories) may inform clinical decisions on how best to treat individuals to reduce fracture risk. © 2019 American Society for Bone and Mineral Research.

Femoral Neck External Size but not aBMD Predicts Structural and Mass Changes for Women Transitioning Through Menopause.

The impact of adult bone traits on changes in bone structure and mass during aging is not well understood. Having shown that intracortical remodeling correlates with external size of adult long bones led us to hypothesize that age-related changes in bone traits also depend on external bone size. We analyzed hip dual-energy X-ray absorptiometry images acquired longitudinally over 14 years for 198 midlife women transitioning through menopause. The 14-year change in bone mineral content (BMC, R2 = 0.03, p = 0.015) and bone area (R2 = 0.13, p = 0.001), but not areal bone mineral density (aBMD, R2 = 0.00, p = 0.931) correlated negatively with baseline femoral neck external size, adjusted for body size using the residuals from a linear regression between baseline bone area and height. The dependence of the 14-year changes in BMC and bone area on baseline bone area remained significant after adjusting for race/ethnicity, postmenopausal hormone use, the 14-year change in weight, and baseline aBMD, weight, height, and age. Women were sorted into tertiles using the baseline bone area-height residuals. The 14-year change in BMC (p = 0.009) and bone area (p = 0.001) but not aBMD (p = 0.788) differed across the tertiles. This suggested that women showed similar changes in aBMD for different structural and biological reasons: women with narrow femoral necks showed smaller changes in BMC but greater increases in bone area compared to women with wide femoral necks who showed greater losses in BMC but without large compensatory increases in bone area. This finding is opposite to expectations that periosteal expansion acts to mechanically offset bone loss. Thus, changes in femoral neck structure and mass during menopause vary widely among women and are predicted by baseline external bone size but not aBMD. How these different structural and mass changes affect individual strength-decline trajectories remains to be determined. © 2017 American Society for Bone and Mineral Research.

A brief intervention to minimize psychosexual morbidity in dyads coping with breast cancer.

PURPOSE/OBJECTIVES: To develop and evaluate the feasibility of a brief intervention to attenuate the incidence of psychosexual morbidity within the dyad secondary to the diagnosis and treatment of breast cancer. DESIGN: Quasiexperimental, including intervention and treatment-as-usual comparison groups. SETTING: Breast clinic of a comprehensive cancer center in the Midwest United States. SAMPLE: 65 recently diagnosed breast cancer survivors who were pre- or perimenopausal and aged 20-55 years, and their partners. METHODS: Three intervention sessions were delivered based on a manual developed for the study. Twenty-five dyads received treatment as usual, 26 dyads received a face-to-face intervention, and 14 dyads received the same intervention by telephone. Questionnaires were completed at baseline, following completion of the intervention, six months postintervention, and from the comparison group at equivalent data points. MAIN RESEARCH VARIABLES: Intimacy, sexual functioning, and dyadic adjustment. FINDINGS: About 98% of dyads completed all intervention sessions, with an equal level of satisfaction among those in the telephone and face-to-face groups. Interesting trends in differences between the intervention and comparison groups on the relationship variables of intimacy, sexual functioning, and dyadic adjustment were obtained; however, given the sample size, power was not sufficient to reach statistical significance. CONCLUSIONS: The intervention is feasible and acceptable for dyads comfortable discussing their relationship. Intervention by telephone was demonstrated to be as effective as the face-to-face mode of delivery. IMPLICATIONS FOR NURSING: Nurses need to provide an opportunity for women to discuss problems they are experiencing relative to sexuality, intimacy, and body image.

Use of hypothermia to allow low-tidal-volume ventilation in a patient with ARDS.

Low-tidal-volume ventilation reduces mortality in patients with ARDS, but there are often challenges in implementing lung-protective ventilation, such as acidosis from hypercapnia. In a patient with severe ARDS we achieved adequate ventilation with a very low tidal volume (4 mL/kg ideal body weight) by inducing mild hypothermia (body temperature 35-36°C).

Statistical analysis and compound selection of combinatorial libraries for soluble epoxide hydrolase.

Inhibitors of soluble epoxide hydrolase (sEH) have been extensively pursued as antihypertensive therapies as well as potential treatment for other cardiovascular dysfunctions and prevention of renal damage. In this study we report quantitative structure-activity relationship (QSAR) models for 1223 structurally diverse sEH inhibitors produced by combinatorial library design and synthesis. Daylight fingerprints, MOE 2D and DragonX descriptors were generated for QSAR modeling approaches. Using these descriptors, a number of statistical models were trained and validated. Of these methods, gradient boosting machines (GBM), partial least-squares (PLS), and Cubist methods demonstrated the best performance on training and test set validation in terms of their leave-group-out cross-validated (LGO-CV) Q(2) and correlation coefficient R(2) (Q(2)(GBM-training) = 0.79, R(2)(GBM-test) = 0.81; Q(2)(PLS-training) = 0.75, R(2)(PLS-test) = 0.75; Q(2)(Cubist-training) = 0.91, R(2)(Cubist-test) = 0.78). A final model was constructed using the consensus approach of the three individual models and showed robust statistics and prediction of the external validation set. The Gaussian process modified sequential elimination of level combinations (G-SELC) method was then used to expand the chemical space beyond what has been explored by combinatorial synthesis. This approach identified 50 new compounds that are structurally diverse and potentially desirable for sEH inhibition based on prior knowledge. The activities of the suggested compounds were then predicted by the consensus QSAR model, and the results supported that the compounds were more likely to exist in the active parts of the chemical space. This study illustrates that the balanced approach by G-SELC could provide a general method for combinatorial library design, to effectively identify promising compounds to be created in the laboratory.

Central venous port placement: a comparison of axillary versus anterior chest wall placement.

BACKGROUND: To compare morbidity in patients with surgically implanted central venous ports that were placed in the subcutaneous tissues of the mid axillary line at the anterior border of the latissimus muscle (group A) versus the anterior chest wall (group C). METHODS: Between 2003 and 2007, a total of 183 patients with breast cancer were taken to the operating room for central venous port placement for delivery of chemotherapy. Port location was determined by patient and surgeon preference. Patient demographics were collected, and complications were evaluated by electronic medical record review. Complications identified included infection, thrombosis, port problems, and catheter problems. Basic descriptive statistics were generated. Patients with axillary ports were then compared to those with chest wall ports by appropriate t-tests or χ(2) tests. RESULTS: During this period, 137 (75%) of 183 ports were placed in the axillary position. The complication rate was 10% (9% in group A vs. 13% in group C). There were 14 catheter-related complications and 5 port-related complications. There was no statistically significant difference in complications between the two groups (P = 0.45). Patients with axillary ports weighed less (mean 75.8 kg in group A vs. 84.9 kg in group C) and were more likely to have catheters placed in the internal jugular vein (82% in group A vs. 56% in group C). CONCLUSIONS: There is no difference in rate or type of complications between axillary and chest wall port location. Subcutaneous ports can safely be placed in the mid axillary line. Axillary ports spare the patient the negative cosmetic outcomes of chest wall ports.

Compliance differences between patients with breast cancer in university and county hospitals.

PURPOSE: Compliance with recommended breast cancer treatments outside the context of a clinical trial differs from that in study populations. The purpose of this study was to examine differences in compliance of breast cancer treatments. PATIENTS AND METHODS: We conducted a retrospective review of 529 patients treated at 2 teaching hospitals in the same city from 2003 to 2006. Compliance with adjuvant therapy recommendations and choice of breast-conserving operations were compared between a university hospital (UH) and a county hospital (CH). RESULTS: The 2 populations demonstrated similar rates of breast conservation (72% vs. 69%). Although use of radiation therapy at the CH was acceptable (82%), patients at the UH were more likely to undergo radiation therapy (95%). The use of hormone therapy was similar at the UH and the CH (> 93%). Patients were more likely to follow physician recommendations for adjuvant chemotherapy at the UH (89%) compared with the CH (70%; P = .0005). Univariate analysis revealed that patient age, tumor size, stage, grade, and estrogen receptor status were all significant predictors of patient compliance with chemotherapy. Preoperative chemotherapy was a strong predictor of patient compliance with chemotherapy (P < .0001). In multivariate analysis, all of the factors predictive of patient compliance in univariate analysis remained significant except tumor grade. CONCLUSION: Preoperative chemotherapy appeared to increase compliance compared with adjuvant chemotherapy in the CH population. Compared with national standards, breast-conserving operations and radiation therapy compliance can be accomplished in an acceptable percentage of underinsured patients.

SLUG/SNAI2 and tumor necrosis factor generate breast cells with CD44+/CD24- phenotype.

BACKGROUND: Breast cancer cells with CD44+/CD24- cell surface marker expression profile are proposed as cancer stem cells (CSCs). Normal breast epithelial cells that are CD44+/CD24- express higher levels of stem/progenitor cell associated genes. We, amongst others, have shown that cancer cells that have undergone epithelial to mesenchymal transition (EMT) display the CD44+/CD24- phenotype. However, whether all genes that induce EMT confer the CD44+/CD24- phenotype is unknown. We hypothesized that only a subset of genes associated with EMT generates CD44+/CD24- cells. METHODS: MCF-10A breast epithelial cells, a subpopulation of which spontaneously acquire the CD44+/CD24- phenotype, were used to identify genes that are differentially expressed in CD44+/CD24- and CD44-/CD24+ cells. Ingenuity pathway analysis was performed to identify signaling networks that linked differentially expressed genes. Two EMT-associated genes elevated in CD44+/CD24- cells, SLUG and Gli-2, were overexpressed in the CD44-/CD24+ subpopulation of MCF-10A cells and MCF-7 cells, which are CD44-/CD24+. Flow cytometry and mammosphere assays were used to assess cell surface markers and stem cell-like properties, respectively. RESULTS: Two thousand thirty five genes were differentially expressed (p < 0.001, fold change >or= 2) between the CD44+/CD24- and CD44-/CD24+ subpopulations of MCF-10A. Thirty-two EMT-associated genes including SLUG, Gli-2, ZEB-1, and ZEB-2 were expressed at higher levels in CD44+/CD24- cells. These EMT-associated genes participate in signaling networks comprising TGFbeta, NF-kappaB, and human chorionic gonadotropin. Treatment with tumor necrosis factor (TNF), which induces NF-kappaB and represses E-cadherin, or overexpression of SLUG in CD44-/CD24+ MCF-10A cells, gave rise to a subpopulation of CD44+/CD24- cells. Overexpression of constitutively active p65 subunit of NF-kappaB in MCF-10A resulted in a dramatic shift to the CD44+/CD24+ phenotype. SLUG overexpression in MCF-7 cells generated CD44+/CD24+ cells with enhanced mammosphere forming ability. In contrast, Gli-2 failed to alter CD44 and CD24 expression. CONCLUSIONS: EMT-mediated generation of CD44+/CD24- or CD44+/CD24+ cells depends on the genes that induce or are associated with EMT. Our studies reveal a role for TNF in altering the phenotype of breast CSC. Additionally, the CD44+/CD24+ phenotype, in the context of SLUG overexpression, can be associated with breast CSC "stemness" behavior based on mammosphere forming ability.

Race and ethnicity and breast cancer outcomes in an underinsured population.

BACKGROUND: The disparity in breast cancer mortality between African American women and non-Hispanic white women has been the subject of increased scrutiny. Few studies have addressed these differences in the setting of equal access to health care. We compared the breast cancer outcomes of underinsured African American and non-Hispanic white patients who were treated at a single institution. METHODS: We conducted a retrospective review of medical records for breast cancer patients who were treated at Wishard Memorial Hospital from January 1, 1997, to February 28, 2006. A total of 574 patients (259 non-Hispanic whites and 315 African Americans) were evaluated. A Cox proportional hazards regression analysis for competing risks was performed. All statistical tests were two-sided. RESULTS: Sociodemographic characteristics were similar in the two groups, and both racial groups were equally unlikely to have undergone screening mammography during the 2 years before diagnosis. Most (84%) of the patients were underinsured. The median time from diagnosis to operation, receipt of adequate surgery, and use of all types of adjuvant therapy were similar in the two groups. Median follow-up was 80.3 months for non-Hispanic whites and 77.9 months for African Americans. After accounting for the effect of comorbidities, African American race was statistically significantly associated with breast cancer-specific mortality (African Americans vs non-Hispanic whites: 26.0% vs 17.5%, P = .028; hazard ratio [HR] of death = 1.64, 95% confidence interval [CI] = 1.06 to 2.55). Adjustment for age at diagnosis, clinical stage, and hormone receptor status attenuated the effect, and the effect of race on breast cancer-specific survival was no longer statistically significant (HR of death from breast cancer = 1.43, 95% CI = 0.89 to 2.30). After adjustment for sociodemographic factors, the hazard ratio for race was further attenuated (HR = 1.26; 95% CI = 0.79 to 2.00). CONCLUSIONS: In this underinsured population, African American patients had poorer breast cancer-specific survival than non-Hispanic white patients. After adjustment for clinical and sociodemographic factors, the effect of race on survival was no longer statistically significant.

The effect of dedicated breast surgeons on the short-term outcomes in breast cancer.

OBJECTIVE: The impact of breast surgeons on short-term outcomes in breast cancer care was compared at a single institution. SUMMARY BACKGROUND DATA: Many studies have demonstrated a correlation between high procedural volume and lower mortality in technically challenging procedures. Breast cancer treatment has significant impact on patient behavior, psychology, and appearance. Therefore, evaluation of outcomes cannot be limited to only operative mortality and morbidity. We sought to determine the effect of dedicated breast cancer surgeons on short-term outcomes at a single institution. METHODS: Wishard Memorial Hospital is the county hospital affiliated with the Indiana University School of Medicine. A retrospective review was performed of all patients from January 1, 1997, to February 28, 2006. On July 1, 2003, coverage for the Breast Clinic was changed from general surgeons (G) to breast surgeons (B). There were 596 patients included in the study period. RESULTS: There were no significant differences in patient demographics or disease characteristics between the 2 time periods. For early stage (stage I and II) breast cancer, a higher percentage of patients underwent breast conservation in the breast surgeon period than in the general surgeon period (P = 0.04). Lumpectomy margins in breast conserving operations during the G period were more often positive (P = 0.025) or close (<1 mm) (P = 0.01). Similarly, the rates of re-excision lumpectomy were also significantly lower during the B period (21% vs. 39%, respectively, P = 0.01). Breast surgeons were more likely to perform the sentinel node procedure (P = 0.001). There were no differences in the use of adjuvant chemotherapy and radiation therapy. The use of hormonal manipulation, however, was significantly higher in the B group than in the G group (P < 0.0002). CONCLUSIONS: Surgeons specialized in diseases of the breast demonstrate significant improvement in short-term outcomes associated with breast cancer treatment at a single institution. The differences identified cannot be attributed to differences in institutional function, patient population, surgeon case volume, or on the influence of nonsurgeon physicians.

Efficacy of the Ahmed S2 glaucoma valve compared with the Baerveldt 250-mm2 glaucoma implant.

OBJECTIVE: To compare the efficacy of the Ahmed S2 Glaucoma Valve with the Baerveldt 250-mm(2) Glaucoma Implant in the treatment of adult glaucoma. DESIGN: Comparative case series. PARTICIPANTS: Fifty-nine eyes of 59 patients who received the Ahmed S2 Glaucoma Valve and 133 eyes of 133 patients who received the Baerveldt 250-mm(2) Glaucoma Implant by the Indiana University Glaucoma Service from 1996 to 2003. METHODS: Eyes that had previous drainage implant procedures were excluded from both groups. If both eyes of a single patient received an implant, the second eye to undergo implantation was excluded from the study. MAIN OUTCOME MEASURES: Kaplan-Meier survival with success defined as intraocular pressure (IOP) > 5 mmHg and < 22 mmHg and at least 20% reduction from preoperative IOP (with or without antiglaucoma medications) and without loss of light perception. Secondary outcome measures included intraocular pressure, visual acuity, number of glaucoma medications, and surgical complications. RESULTS: The 2 groups were similar with regards to age, gender, race, neovascular glaucoma diagnosis, number of prior ocular surgeries, preoperative IOP, and number of preoperative glaucoma medications. Mean durations of follow-up were 20.0 months for Ahmed eyes and 22.9 months for Baerveldt eyes. Cumulative successes in the Ahmed group were 0.73 at 1 year and 0.62 at 2 years, whereas cumulative successes in the Baerveldt group were 0.92 at 1 year and 0.85 at 2 years (Kaplan-Meier survival functions: P = 0.03, log rank test). Male gender, African descent, neovascular glaucoma, and Ahmed implantation were found to be significant predictors of failure. At last follow-up visit, eyes in the Ahmed group had a significantly higher mean IOP (19.8+/-9.5 vs. 15.8+/-7.9 mmHg, P = 0.003, t test) and more antiglaucoma medications (1.4+/-1.2 vs. 0.9+/-1.1 medications, P = 0.008, Mann-Whitney test) than eyes in the Baerveldt group. Two methods for avoiding hypotony after Baerveldt 250-mm(2) implantation had similar outcomes. CONCLUSIONS: Our study suggests that the Ahmed S2 Glaucoma Valve may be less effective at lowering IOP than the Baerveldt 250-mm(2) Glaucoma Implant.

MOZ and MOZ-CBP cooperate with NF-kappaB to activate transcription from NF-kappaB-dependent promoters.

OBJECTIVE: Monocytic zinc finger (MOZ) maintains hematopoietic stem cells and, upon fusion to the coactivator CREB-binding protein (CBP), induces acute myeloid leukemia (AML). Leukemic stem cells in AML often exhibit excessive signal-dependent activity of the transcription factor nuclear factor (NF)-kappaB. Because aberrant interaction between NF-kappaB and coactivators represents an alternative mechanism for enhancing NF-kappaB activity, we evaluated whether MOZ and MOZ-CBP cooperate with NF-kappaB to activate transcription from NF-kappaB-dependent promoters. METHODS: The ability of MOZ, MOZ mutants, and MOZ-CBP to enhance expression of NF-kappaB-dependent promoters was tested in reporter studies. The interaction between MOZ and NF-kappaB was evaluated by both coimmunoprecipitation and glutathione S-transferase pulldown assays. RESULTS: MOZ activates transcription from the NF-kappaB-dependent interleukin-8 promoter; interestingly, this effect is markedly enhanced by CBP. Although MOZ has less potent transcriptional activity than MOZ-CBP, both proteins cooperate with steroid receptor coactivator-1 to activate transcription. MOZ also induces multiple NF-kappaB-dependent viral promoters. Importantly, MOZ associates in a protein complex with the p65 subunit of NF-kappaB and interacts directly with p65 in vitro. Transcriptional activity of MOZ requires its C-terminal domain, which is absent from MOZ-CBP, indicating that the transcriptional activity of MOZ-CBP derives from its CBP sequence. CONCLUSIONS: MOZ interacts with the p65 subunit of NF-kappaB and enhances expression of NF-kappaB-dependent promoters. The more potent transcriptional activity of MOZ-CBP derives from its CBP sequence. Thus, interaction between NF-kappaB and MOZ-CBP may play an important role in the pathogenesis of certain acute myeloid leukemias.