Quantitative evaluation of DNA hypermethylation in malignant and benign breast tissue and fluids.

The assessment of DNA had demonstrated altered methylation in malignant compared to benign breast tissue. The purpose of our study was to (i) confirm the predictive ability of methylation assessment in breast tissue, and (ii) use the genes found to be cancer predictive in tissue to evaluate the diagnostic potential of hypermethylation assessment in nipple aspirate fluid (NAF) and mammary ductoscopic (MD) samples. Quantitative methylation specific (qMS)-PCR was conducted on three specimen sets: 44 malignant (CA) and 34 normal (NL) tissue specimens, 18 matched CA, adjacent normal (ANL) tissue and NAF specimens, and 119 MD specimens. Training and validation tissue sets were analyzed to determine the optimal group of cancer predictive genes for NAF and MD analysis. NAF and MD cytologic review were also performed. Methylation of CCND-2, p16, RAR-beta and RASSF-1a was significantly more prevalent in tumor than in normal tissue specimens. Receiver operating characteristic curve analysis demonstrated an area under the curve of 0.96. For the 18 matched CA, ANL and NAF specimens, the four predictive genes identified in cancer tissue contained increased methylation in CA vs. ANL tissue; NAF samples had higher methylation than ANL specimens. Methylation frequency was higher in MD specimens from breasts with cancer than benign samples for p16 and RASSF-1a. In summary, i) routine quantitative DNA methylation assessment in NAF and MD samples is possible, and ii) genes hypermethylated in malignant breast tissue are also altered in matched NAF and in MD samples, and may be useful to assist in early breast cancer detection.

Carbohydrate antigens in nipple aspirate fluid predict the presence of atypia and cancer in women requiring diagnostic breast biopsy.

BACKGROUND: The goal of this prospective study was to determine (a) concentrations of the carbohydrate biomarkers Thomsen Friedenreich (TF) antigen and its precursor, Tn antigen, in nipple discharge (ND) collected from women requiring biopsy because of a suspicious breast lesion; and (b) if concentration levels predicted pathologic diagnosis. METHODS: Adult women requiring biopsy to exclude breast cancer were enrolled and ND obtained. The samples from 124 women were analyzed using an anti-TF and anti-Tn monoclonal antibodies in direct immunoassay. RESULTS: The highest median concentration in ND for TF and Tn was in women with ductal carcinoma in situ (DCIS). TF was higher in women with 1) cancer (DCIS or invasive) vs. either no cancer (atypia or benign pathology, p = .048), or benign pathology (p = .018); and 2) abnormal (atypia or cancer) versus benign pathology (p = .016); and was more predictive of atypia or cancer in post- compared to premenopausal women. Tn was not predictive of disease. High TF concentration and age were independent predictors of disease, correctly classifying either cancer or abnormal vs. benign pathology 83% of the time in postmenopausal women. CONCLUSIONS: TF concentrations in ND were higher in women with precancer and cancer compared to women with benign disease, and TF was an independent predictor of breast atypia and cancer. TF may prove useful in early breast cancer detection.

Soy isoflavones have an antiestrogenic effect and alter mammary promoter hypermethylation in healthy premenopausal women.

We determined if soy isoflavones have dose-related estrogenic and methylation effects. Thirty-four healthy premenopausal women were randomized to 40 mg or 140 mg isoflavones daily through one menstrual cycle. Breast specific and systemic estrogenic effects were assessed measuring the estrogenic marker complement (C)3 and changes in cytology, whereas methylation assessment of 5 cancer related genes (p16, RASSF1A, RARbeta2, ER, and CCND2) was performed on intraductal specimens. Serum genistein significantly increased after consuming both isoflavone doses. Cytology did not significantly change at either isoflavone dose. Serum C3 levels posttreatment were inversely related to change in serum genistein (r =-0.76, P = 0.0045) in women consuming low but not high dose isoflavones. The RAR beta 2 hypermethylation increase posttreatment correlated with the posttreatment genistein level considering the entire group (r = 0.67, P = 0.0017) and those receiving high-dose isoflavones (r = 0.68, P = 0.021). At the low but not the high isoflavone dose, CCND2 hypermethylation increase correlated with posttreatment genistein levels (r = 0.79, P = 0.011). In summary, the inverse correlation between C3 and genistein suggests an antiestrogenic effect. Isoflavones induced dose-specific changes in RARbeta2 and CCND2 gene methylation, which correlated with genistein levels. This work provides novel insights into estrogenic and methylation effects of dietary isoflavones.

Celecoxib concentration predicts decrease in prostaglandin E2 concentrations in nipple aspirate fluid from high risk women.

BACKGROUND: Epidemiologic studies suggest that long term low dose celecoxib use significantly lowers breast cancer risk. We previously demonstrated that 400 mg celecoxib taken twice daily for 2 weeks lowered circulating plasma and breast nipple aspirate fluid (NAF) prostaglandin (PG)E2 concentrations in post- but not premenopausal high risk women. We hypothesized that circulating concentrations of celecoxib influenced PGE2 response, and that plasma levels of the drug are influenced by menopausal status. To address these hypotheses, the aims of the study were to determine: 1) if circulating plasma concentrations of celecoxib correlated with the change in plasma or NAF PGE2 concentrations from baseline to end of treatment, and 2) whether menopausal status influenced circulating levels of celecoxib. METHODS: Matched NAF and plasma were collected from 46 high risk women who were administered celecoxib twice daily for two weeks, 20 subjects receiving 200 mg and 26 subjects 400 mg of the agent. NAF and plasma samples were collected before and 2 weeks after taking celecoxib. RESULTS: In women taking 400 mg bid celecoxib, plasma concentrations of the agent correlated inversely with the change in NAF PGE2 levels from pre- to posttreatment. Nonsignificant trends toward higher celecoxib levels were observed in post- compared to premenopausal women. There was a significant decrease in NAF but not plasma PGE2 concentrations in postmenopausal women who took 400 mg celecoxib (p = 0.03). CONCLUSION: In high risk women taking 400 mg celecoxib twice daily, plasma concentrations of celecoxib correlated with downregulation of PGE2 production by breast tissue. Strategies synergistic with celecoxib to downregulate PGE2 are of interest, in order to minimize the celecoxib dose required to have an effect.

uPA is upregulated by high dose celecoxib in women at increased risk of developing breast cancer.

BACKGROUND: While increased urokinase-type plasminogen activator (uPA) expression in breast cancer tissue is directly associated with poor prognosis, recent evidence suggests that uPA overexpression may suppress tumor growth and prolong survival. Celecoxib has been shown to have antiangiogenic and antiproliferative properties. We sought to determine if uPA, PA inhibitor (PAI)-1 and prostaglandin (PG)E2 expression in nipple aspirate fluid (NAF) and uPA and PGE2 expression in plasma were altered by celecoxib dose and concentration in women at increased breast cancer risk. METHODS: NAF and plasma samples were collected in women at increased breast cancer risk before and 2 weeks after taking celecoxib 200 or 400 mg twice daily (bid). uPA, PAI-1 and PGE2 were measured before and after intervention. RESULTS: Celecoxib concentrations trended higher in women taking 400 mg (median 1025.0 ng/mL) compared to 200 mg bid (median 227.3 ng/mL), and in post- (534.6 ng/mL) compared to premenopausal (227.3 ng/mL) women. In postmenopausal women treated with the higher (400 mg bid) celecoxib dose, uPA concentrations increased, while PAI-1 and PGE2 decreased. In women taking the higher dose, both PAI-1 (r = -.97, p = .0048) and PGE2 (r = -.69, p = .019) in NAF and uPA in plasma (r = .45, p = .023) were correlated with celecoxib concentrations. CONCLUSION: Celecoxib concentrations after treatment correlate inversely with the change in PAI-1 and PGE2 in the breast and directly with the change in uPA in the circulation. uPA upregulation, in concert with PAI-1 and PGE2 downregulation, may have a cancer preventive effect.

Prospective multicenter trial to determine the feasibility of collection and predictive ability of breast fluid analysis in postmenopausal women receiving SERMs.

We conducted a prospective ancillary study to the study of tamoxifen and raloxifene (STAR) trial involving 4 institutions to determine: (1) our ability to collect nipple aspirate fluid (NAF) in postmenopausal women taking medication to decrease breast proliferation and (2) NAF biomarkers associated with breast cancer. NAF was collected before and 6 months after starting treatment. Three biomarkers, cytology, fluid volume, and prostate-specific antigen (PSA), were analyzed. NAF was collected from 26 of 33 (79%) enrolled subjects at baseline. We were able to collect fluid in 84% of the subjects who produced NAF at baseline and returned for second aspiration after 6 months of treatment. In these women, cytology was unchanged in 85%, improved in 11%, and worsened in 4% of breasts. Median PSA increased from 37.5 to 112 ng/L after treatment. NAF volume did not significantly change. In conclusion, after treatment with tamoxifen or raloxifene, changes in both NAF cytology and PSA were generally favorable, consistent with their expected antiproliferative effective effect on the breast. Multiinstitutional clinical trials in postmenopausal women using NAF biomarkers as the primary endpoints are feasible.

Rural hospital patient safety systems implementation in two States.

CONTEXT AND PURPOSE: With heightened attention to medical errors and patient safety, we surveyed Utah and Missouri hospitals to assess the "state of the art" in patient safety systems and identify changes over time. This study examines differences between urban and rural hospitals. METHODS: Survey of all acute care hospitals in Utah and Missouri at 2 points in time (2002 and 2004). Factor analysis was used to develop 7 latent variables to summarize the data, comparing rural and urban hospitals at each point in time and on change between the 2 survey times. FINDINGS: On 3 of the 7 latent variables, there was a statistically significant difference between rural and urban hospitals at the first survey, with rural hospitals indicating lower levels of implementation. The differences remained present on 2 of those latent variables at the second survey. In both cases, 1 of those variables was computerized physician order entry (CPOE) systems. Rural hospitals reported more improvement in systems implementation between the 2 survey times, with the difference statistically significant on 1 of the 7 latent variables; the greatest improvement was in implementation of "root cause analysis." CONCLUSIONS: Adoption of patient safety systems overall is low. Although rates of adoption among rural versus urban hospitals appear lower, most differences are not statistically significant; the gap between rural and urban hospitals relative to quality measures is narrowing. Change in rural and urban hospitals is in the right direction, with the rate of change higher in rural hospitals for many systems.

Hospital patient safety: characteristics of best-performing hospitals.

Hospitals have made slow progress in meeting the Institute of Medicine's patient safety goals, and implementation of safety systems has been inconsistent. The next logical question is this: What organizational characteristics predict greater implementation of patient safety systems, in terms of both extent of systems and progress over time? To answer this question, a survey was administered to 107 hospitals at two points in time. Data were consolidated into seven latent variables measuring progress in specific areas. Using the overall measure, Joint Commission-accredited hospitals showed statistically significant improvement, as reflected in the sum score (p = .01); nonaccredited hospitals did not show statistically significant improvement (p = .21). Joint Commission accreditation was the key predictor of patient safety system implementation. Management type and urban/rural status were secondary predictors. Several factors may account for the strong association between accreditation and patient safety system implementation. In 2003, the Joint Commission began tying accreditation to patient safety goals. Also, Joint Commission data are now widely available to the public and may stimulate hospitals to address safety issues. Healthcare executives, hospital trustees, regulators, and policymakers should encourage Joint Commission accreditation and reward hospital efforts toward meeting Joint Commission standards. The Joint Commission should continually strive to maintain evidence-based and state-of-the-art standards that advance the aim of providing the best possible care for hospitalized patients.

Celecoxib decreases prostaglandin E2 concentrations in nipple aspirate fluid from high risk postmenopausal women and women with breast cancer.

BACKGROUND: Celecoxib inhibits PGE2 production in cancerous tissue. We previously reported that PGE2 levels in nipple aspirate fluid (NAF) and plasma were not decreased in women at increased breast cancer risk who received celecoxib 200 mg twice daily (bid). The endpoints of the current study were to determine if a short course of celecoxib 400 mg bid would decrease PGE2 levels in women 1) at increased breast cancer risk, and 2) with established breast cancer. METHODS: NAF and plasma samples were collected before, 2 weeks after taking celecoxib 400 mg bid, and two weeks after washout from 26 women who were at increased breast cancer risk. From 13 women with newly diagnosed breast cancer, NAF from the incident breast and plasma were collected before and on average 2 weeks after taking celecoxib. Additionally, in nine of the 13 women with breast cancer, NAF was collected from the contralateral breast. RESULTS: No consistent change in NAF or plasma PGE2 levels was noted in high risk premenopausal women. NAF PGE2 levels decreased after celecoxib administration in postmenopausal high risk women (p = 0.02), and in both the NAF (p = 0.02) and plasma (p = 0.03) of women with breast cancer. CONCLUSION: Celecoxib 400 mg bid taken on average for 2 weeks significantly decreased NAF, but not plasma, PGE2 levels in postmenopausal high risk women, and decreased both NAF and plasma PGE2 levels in women with newly diagnosed breast cancer. PGE2 levels may predict celecoxib breast cancer prevention and treatment efficacy. Our observations are preliminary, and larger studies to confirm and extend these findings are warranted.

The long road to patient safety: A status report on Missouri acute-care hospitals.

We surveyed Missouri's acute-care hospitals twice to assess the "state of the art" in patient safety systems. We found improvements in all areas studied, but progress is modest and falls short of national recommendations. We urge all Missourians to become more knowledgeable and assertive consumers of hospital care, and we ask them to support Missouri hospitals in their patient safety efforts, including provision of financial resources to put necessary systems in place.

Characteristics of smoking cessation guideline use by primary care physicians.

This study, based on a random sample mail survey of Missouri primary care physicians, identifies: (1) the extent and predictors of these physicians' awareness, agreement, adoption, and adherence to tobacco cessation guidelines; and (2) their knowledge/ beliefs about tobacco interventions. While primary care physicians play a key role in helping patients quit smoking, guidelines are not widely followed. "Reliance on respected colleagues for advice" was the one variable consistently identified as a predictor of guideline compliance.

The long road to patient safety: a status report on patient safety systems.

CONTEXT: Since the Institute of Medicine (IOM) reports on medical errors and quality, national attention has focused on improving patient safety through changes in "systems" of care. These reports resulted in a new paradigm that, rather than centering on individual errors, focuses on the "systems" necessary to facilitate and enhance quality and protect patients. OBJECTIVES: To assess the status of hospital patient safety systems since the release of the IOM reports and to identify changes over time in 2 states that collaborated on a patient safety project funded by the Agency for Healthcare Research and Quality. DESIGN, SETTING, AND PARTICIPANTS: Survey of all acute care hospitals in Missouri and Utah at 2 points in time, in 2002 and 2004, using a 91-item comprehensive questionnaire (n = 126 for survey 1 and n = 128 for survey 2). To assess changes over time, we also studied the cohort of 107 hospitals that responded to both surveys. MAIN OUTCOME MEASURES: Responses to the 91-question survey as well as changes in responses to the survey questions over an 18-month period. Seven latent variables were constructed to represent the most important patient safety constructs studied: computerized physician order entry systems, computerized test results, and assessments of adverse events; specific patient safety policies; use of data in patient safety programs; drug storage, administration, and safety procedures; manner of handling adverse event/error reporting; prevention policies; and root cause analysis. For each hospital, the 7 latent variables were summed to give an overall measure of the patient safety status of the hospital. RESULTS: Development and implementation of patient safety systems is at best modest. Self-reported regression in patient safety systems was also found. While 74% of hospitals reported full implementation of a written patient safety plan, nearly 9% reported no plan. The area of surgery appears to have the greatest level of patient safety systems. Other areas, such as medications, with a long history of efforts in patient safety and error prevention, showed improvements, but the percentage of hospitals with various safety systems was already high at baseline for many systems. Some findings are surprising, given the overall trends; for example, while a substantial percentage of hospitals have medication safety systems, only 34.1% [corrected] reported full implementation at survey 2 of computerized physician order entry systems for medications, despite the growth of computer technology in general and in hospital billing systems in particular. CONCLUSIONS: The current status of hospital patient safety systems is not close to meeting IOM recommendations. Data are consistent with recent reports that patient safety system progress is slow and is a cause for great concern. Efforts for improvement must be accelerated.

Self-efficacy and pain behavior among subjects with fibromyalgia.

Given the lack of objective physical measures for assessing fibromyalgia syndrome (FS), the role of pain assessment is particularly important. The role of psychological factors is controversial among FS patients. This study was designed to better understand the relationship between pain behaviors and psychological variables. Specifically, this study (1) refined a pain behavior observation (PBO) methodology for use with FS patients, (2) determined whether stretching is a valid pain behavior, and (3) assessed whether psychological variables including self-efficacy and/or depression can predict pain behaviors after controlling for disease severity and age. The 73 FS subjects meeting the American College of Rheumatology classification system completed questionnaires measuring self-efficacy, depression, and pain. Trained physicians conducted tender-point examinations. Subjects were video-taped using a standardized procedure. Two trained raters independently coded all pain behaviors. Kappa coefficients and correlations among pain behaviors and self-reported pain indicated that the PBO method was both reliable and valid. However, the newly defined pain behavior 'stretching' was found to be negatively associated with self-reported pain. Hierarchical multiple regression (MR) analyses revealed that depression did not predict pain behavior over and above myalgic scores and age; however, in 3 separate MR analyses, self-efficacy for function, pain, and other symptoms each predicted pain behavior over and above myalgic scores and age. This study indicated that the original pain behavior scoring methodology is appropriate for use with the FS population and should not be modified to include the pain behavior 'stretching'. Self-efficacy was related to pain behavior while depression was not among this FS sample.

The association of bloody nipple discharge with breast pathology.

BACKGROUND: It is believed that bloody spontaneous nipple discharge (SND) portends a greater chance of malignancy than nonbloody discharge, and that cytologic evaluation of SND assists in treatment planning. Our aims were to assess (1) the pathology of women with/without SND who require diagnostic breast surgery, (2) whether bloody SND is associated with a different spectrum of pathologic findings than nonbloody, and (3) whether SND cytology is influenced by pathologic findings. METHODS: One hundred seventy-five women who underwent breast operation were enrolled. Cytologic evaluation was performed on Papanicolaou-stained cytospin preparations of SND. RESULTS: Papilloma and hyperplasia (both without atypia) were more frequent in breasts with than without SND; breast cancer was more common in breasts without SND (P <.001 for all). All cases of breast cancer with SND, but only 33% without, contained a papilloma or papillary features. Seventy-five percent of cancers presenting with SND were nonbloody. Papilloma was the most common diagnosis in breasts with bloody SND and was more common (P=.017) than in breasts without, whereas hyperplasia was the most common diagnosis in breasts with nonbloody SND and was more common (P=.031) than in breasts with bloody SND. SND cytology was not significantly influenced by pathology. CONCLUSIONS: Breast cancer can present as unilateral nonbloody SND, indicating the importance of surgical intervention. Papilloma was more common in women with SND than without and most often presented as bloody SND.

Health care system and insurer support for smoking cessation guideline implementation.

Physician use of clinical practice guidelines (CPGs) is disappointingly low in the United States. Much emphasis historically has been placed on the individual clinician to implement use of guidelines in practice. Recently, the Public Health Service issued an updated set of smoking cessation guidelines that include recommendations not only for patients and physicians, but also for health care administrators, insurers, and purchasers. A random sample of Missouri family physicians and general internists was used to determine, for the first time empirically, whether physicians receiving external support for guideline implementation were more likely to adopt and adhere to guidelines in practice. Fewer than 20 percent of physicians receive system support consistent with the updated guideline for smoking cessation. Only 32 percent of physicians who are unaware of the guidelines receive any of the recommended external support, while nearly 60 percent of physicians who adhere to the guidelines in practice are receiving some form of external support. Thus, the fundamental issue that requires national attention is that successful guideline implementation is highly dependent on administrative supports from health care organizations and insurers.

Quality of intrathecal baclofen from different sources.

OBJECTIVE: To compare the quality of intrathecal baclofen obtained from a national compounding pharmacy (AnazaoHealth) with the manufactured product (Lioresal) with regard to accuracy and precision of baclofen concentration, and the content of the baclofen degradation product, 4-(4-chlorophenyl)-2-pyrrolidinone (PYR). DESIGN: Samples of baclofen used for refilling intrathecal pumps were placed in 1.2-mL silicone gasket-sealed cryogenic vials and stored at or lower than -25°C. Each sample was a different lot number (Lioresal) or prescription number (AnazaoHealth). The laboratory was blinded to the source of the solutions. Coupled with electrospray ionization-mass spectrometry analyte confirmation, quantitation of baclofen and PYR in each sample was performed in duplicate by using high-performance liquid chromatography with ultraviolet detection via a photodiode array detector. SETTING: Outpatient clinic. PARTICIPANTS: Patients with intrathecal baclofen pumps. MAIN OUTCOME MEASURES: Accuracy and precision of baclofen concentration, and concentration of PYR. RESULTS: The difference of mean concentrations from expected concentrations of 500 and 2000 µg/mL were significantly greater for samples from AnazaoHealth compared with Lioresal. Values are shown as mean (± standard error), with n the number samples, for AnazaoHealth compared with Lioresal: (537.1 ± 6.7 µg/mL [n = 8] versus 515.6 ± 0.82 µg/mL [n = 5]; P = .034, respectively) and (2211.4 ± 21.6 µg/mL [n = 12] versus 2055.3 ± 8.7 µg/mL [n = 4]; P = .004, respectively). AnazaoHealth samples with expected concentration of 4000 µg/mL were 3987.7 ± 79.9 µg/mL, n = 7. All 9 Lioresal samples were within 5% of the expected concentration. Of 27 AnazaoHealth samples, 22 were more than 5%, and 8 were more than 10% different from the expected concentration. No PYR was detected in any sample from AnazaoHealth. All samples of Lioresal contained PYR, but all solutions contained less PYR than 1% of the baclofen concentration. CONCLUSIONS: Lioresal was more accurate in concentration and more precise among batches than compounded intrathecal baclofen but had higher levels of PYR.

The research mentoring relationship in family medicine: findings from the grant generating project.

BACKGROUND AND OBJECTIVES: Mentoring has been acknowledged as a critical factor in the development of family medicine academicians. Specific aims were to describe the research mentoring in family medicine from the experience of both mentors and protégés and identify characteristics that mentors and protégés associated with a successful mentoring relationship. The Grant Generating Project (GGP) Fellowship, a training and mentoring program for family medicine researchers, provided a natural opportunity to study these issues and better understand what is successful in research mentoring. METHODS: Separate mentor and protégés surveys measured perceptions about the extent of mentoring assistance, perceived relationship success, costs and benefits of the relationship, and the nature and duration of the relationship. Correlations between demographic characteristics and the mentoring relationship were also examined. RESULTS: Mentors were generally professors (78%), male (82%), with a mean age of 53 years, while protégés were assistant professors (53%) and almost evenly divided between male (51%) and female (49%) with mean age of 44 years. Both mentors and protégés describe the mentoring relationship in general to be of benefit to both mentor and protégé. Nonetheless, statistically significant differences between mentor-protégé responses were found for nine of the 20 survey items. Mentors tended to give higher values in their ratings of specific mentor-protégé relationship variables. Significant positive correlations were found between benefit, quality of the relationship, and mentoring assistance and the number of hours per month of mentor-protégé interaction, the number of mentor-protégé meetings per month, and the number of months the mentor worked with the protégé. Mentor-protégé acquaintance before the GGP fellowship was significantly correlated with cost, benefit, and mentoring assistance. CONCLUSIONS: This study shows agreement between mentor and protégé regarding the mentors' ability to promote the protégés, provide important technical skills, convey respect for the protégés, and serve as a friend and role model. Protégés tend to be more connected with their colleagues and with their profession, perhaps in part because the mentoring relationship facilitates networking opportunities provided by the mentor. In particular, excellent mentors can provide protégés with opportunities to meet other influential scholars at conferences and/or through various forms of correspondence. Such relationships can be helpful to the protégé in developing a constellation of mentoring relationships that may result in more successful research careers. Future studies should examine the relationship upon various outcomes.

Percent true calcium absorption, mineral metabolism, and bone mass in children with arthritis: effect of supplementation with vitamin D3 and calcium.

OBJECTIVE: To assess whether percent true calcium absorption (alpha) is normal and whether supplementation with placebo, vitamin D3 (2,000 IU/day), calcium (1,000 mg/day), or vitamin D3 plus calcium improves alpha, mineral metabolism, or bone mass accrual in children with arthritis. METHODS: Eighteen children received all 4 treatments, each for 6 months, in 4 different, randomly assigned orders. Changes in levels of 25-hydroxyvitamin D (25[OH]D), 1,25-dihydroxyvitamin D (1,25[OH]2D), parathyroid hormone, bone turnover markers, and minerals and in bone mineral content were measured. Calcium absorption was determined with a dual stable isotope method using 48Ca administered intravenously and 46Ca administered orally, and measuring 48Ca, 46Ca, and 42Ca in a 24-hour urine specimen by high-resolution inductively coupled plasma mass spectroscopy. Wilcoxon's signed rank test was used both to identify significant change over the treatment period with a given regimen and to compare change with an experimental treatment versus change with placebo. RESULTS: Percent true calcium absorption was in the lower-normal range and did not differ by treatment (mean+/-SD 28.3+/-20.2% with placebo, 26.1+/-12.1% with calcium, 19.2+/-11.7% with vitamin D3, and 27.1+/-16.5% with vitamin D3 plus calcium). With vitamin D3 and vitamin D3 plus calcium treatment, 25(OH)D levels were increased and 1,25(OH)2D levels were maintained. Serum calcium levels were increased only with vitamin D3 and vitamin D3 plus calcium treatment. Levels of bone turnover markers and increases in bone mineral content did not differ by treatment. CONCLUSION: The findings of this study indicate that percent true calcium absorption is low-normal in children with arthritis. Vitamin D3 at 2,000 IU/day increases serum 25(OH)D and calcium levels but does not improve bone mass accretion. Calcium at 1,000 mg/day also failed to improve bone mass.

Percent true calcium absorption, mineral metabolism, and bone mineralization in children with cystic fibrosis: effect of supplementation with vitamin D and calcium.

OBJECTIVE: To assess whether percent true calcium absorption (alpha) is normal in children with cystic fibrosis (CF) and to assess whether supplementation with 2,000 IU vitamin D(3), 1 g calcium, or both will alter alpha, mineral metabolism, and/or bone mass in children with CF. STUDY DESIGN: Fifteen children ages 7-13 were randomly assigned to one of four different orders to receive all four 6-month treatments including placebos. Change in 25-hydroxyvitamin D (25-OHD), 1,25-dihydroxyvitamin D (1,25(OH)(2)D), PTH, bone turnover markers, and minerals after 6 months, and bone mineral content (Hologic 1000W) after 9 months was measured. alpha was measured by a dual stable isotope method using (48)Ca intravenously and (46)Ca orally and measuring (48)Ca, (46)Ca, and (42)Ca in a 24-hr urine using High Resolution Inductively Coupled Mass Spectroscopy (HR-ICP-MS). Analysis used Wilcoxon Sign Ranks. RESULTS: alpha was in the normal range and did not differ by treatment (P 35 +/- 10%, Ca 38 +/- 23%, D 36 +/- 11%, D + Ca 46 +/- 21%). One gram calcium did not increase serum or urine calcium. Two thousand IU D(3) did not increase 25-OHD or change 1,25(OH)(2)D. Serum and urine minerals, markers of bone turnover and bone mineral gains did not differ by treatment. CONCLUSIONS: alpha is normal in children with CF. One gram calcium and/or 2,000 IU D(3) does not change alpha or increase 25-OHD, serum calcium, or mineralization. Longer trials of a significantly higher dose of vitamin D(3) shown to increase serum 25-OHD are needed to assess effects on mineral metabolism and bone mass accrual. However, study of therapeutic options other than calcium and vitamin D should be encouraged.

Increased expression of the inflammatory protein YKL-40 in precancers of the breast.

Serum levels of YKL-40 have been associated with inflammatory diseases and breast cancer. Our purpose was to determine if YKL-40 in breast tissue, nipple aspirate fluid (NAF) and serum is (i) concentrated in NAF compared to matched serum, (ii) increased in the NAF, serum or tissue of women with biopsy proven precancer or cancer compared to healthy women and (iii) influenced by menopausal status. 118 women (61 healthy subjects, 10 with precancer and 47 with breast cancer) aged 17-95 years provided NAF with or without serum samples for analysis. Matched tissue was analyzed from a subset of subjects who underwent breast biopsy. All NAF and serum samples had detectable levels of YKL-40. Median YKL-40 levels for the entire cohort were 683 fold higher in NAF than serum. Premenopausal subjects had higher NAF and lower serum levels of YKL-40 than postmenopausal subjects. YKL-40 levels in NAF but not serum were higher in women with precancer (atypical hyperplasia and lobular carcinoma in situ) than in either healthy subjects (p = 0.025) or subjects with breast cancer (p = 0.015). In women with precancer, YKL-40 distribution in tissue correlated with YKL serum level (p = 0.043). YKL-40 is concentrated in NAF, with the highest concentrations in premenopausal women. NAF levels of YKL-40 are significantly higher in women with precancers than healthy subjects, suggesting that measuring YKL-40 in NAF may improve the identification of women at increased breast cancer risk.