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Telephone counseling intervention increases intakes of micronutrient- and phytochemical-rich vegetables, fruit and fiber in breast cancer survivors.

Although a large body of evidence suggests that diet may play an important role in cancer prevention, randomized controlled trials reported to date have not achieved sufficient increases in protective micronutrients and phytochemicals to adequately test the hypothesis that diet can reduce cancer risk. The Women's Healthy Eating and Living (WHEL) Study, a randomized controlled trial of the role diet modification may play in future breast cancer events, introduced an innovative theory-based telephone counseling intervention to teach participants to consume a high fiber, low fat diet emphasizing vegetables and fruits rich in carotenoids and other potentially protective phytochemicals. This report examines the baseline to 12-mo changes in dietary intakes of 2970 participants, assessed through 24-h recalls and validated with plasma carotenoid concentrations. At 12 mo, the intervention group reported a significantly increased daily vegetable intake (+vegetable juice) of 7.1 servings (+82%) and fruit intake of 3.9 servings (+18%). Fiber intake increased from 3.04 to 4.16 g/(MJ. d), whereas energy from fat decreased significantly from 28.6 to 23.7%. Plasma carotenoid concentrations increased significantly, i.e., alpha-carotene (+223%); beta-carotene (+87%); lutein (+29%); and lycopene (+17%). In the comparison group, dietary intake and plasma carotenoid concentrations were essentially identical to those of the intervention group at baseline and were unchanged at 12 mo. The WHEL Study showed that a telephone counseling intervention can achieve major increases in micronutrient- and phytochemical-rich vegetables, fruit and fiber intakes, enabling an investigation of the potential cancer preventive effects of these food components.

Adult↗

Left ventricular mechanoenergetics during asynchronous left atrial-to-aortic bypass. Effects of pumping rate on cardiac workload and myocardial oxygen consumption.

The purpose of this study was to analyze left ventricular energetics during asynchronous, pulsatile left atrial to aortic bypass in the failing heart with the use of the pressure-volume relationship. In 12 anesthetized Holstein calves (body weight 94 +/- 7 kg), 10 microns microspheres (3.3 x 10(7) +/- 1.1 x 10(7)/100 gm left ventricular weight) were injected into the left main coronary artery to induce heart failure. Baseline left ventricular end-systolic elastance significantly decreased from 7.9 +/- 0.7 to 5.5 +/- 0.4 mm Hg/ml 100 gm left ventricular weight. Left ventricular pressure was measured with a micromanometer, and ultrasonic dimension transducers measured left ventricular orthogonal diameters. Ellipsoidal geometry was used to calculate simultaneous left ventricular volume. End-systolic elastance, pressure-volume area, external work, potential energy, and myocardial oxygen consumption were analyzed during steady-state contractions. After pre-pulsatile left atrial to aortic bypass measurements were taken, the measurements were repeated during asynchronous pulsatile left atrial to aortic bypass at the maximal pumping rate (69 +/- 13 beats/min) termed 100%, and then 80%, 60%, and 40% of the maximal pumping rate in the full to empty mode. With increases in pumping rate, pressure-volume area and external work proportionally decreased, whereas potential energy remained unchanged except for 100% of maximal pumping rate. Pressure-volume area correlated linearly with myocardial oxygen consumption during asynchronous pulsatile left atrial to aortic bypass (r = 0.971). As a result, pumping rate correlated linearly with conservation of myocardial oxygen consumption (r = 0.998). In conclusion, decreased pressure-volume area accounts for the reduction in myocardial oxygen consumption during asynchronous pulsatile left atrial to aortic bypass. Conservation of myocardial oxygen consumption is mainly attributed to the reduction of external work.

Animals↗

Biomechanical evaluation of dual-energy X-ray absorptiometry for predicting fracture loads of the infant femur for injury investigation: an in vitro porcine model.

OBJECTIVE: The purpose of this study was to determine the ability of bone mineral density (BMD) measured by dual-energy x-ray absorptiometry (DXA) and geometry measured by biplanar x-ray to predict fracture mechanics in vitro in an immature femur model. DESIGN: Prospective analysis of radiographic and biomechanical data was performed. SETTING: In vitro experimentation. INTERVENTIONS: Bone geometry and DXA data were obtained before mechanical testing. Twenty-two porcine femora from males and females (age 3 to 12 months; body weight 3.6 to 7.0 kilograms) were fractured. Mechanical tests were performed on the diaphysis of the femora in two loading configurations: (a) three-point bending to simulate loads that result in transverse fractures; and (b) torsion to simulate twisting injuries that result in spiral fractures. MAIN OUTCOME MEASURES: Correlation of radiographic data with the experimentally determined bone strength. RESULTS: Three-point bending consistently resulted in transverse fractures. Femoral diaphysis BMD (mean, 0.304 grams per square centimeter; SD, 0.028 grams per square centimeter) strongly correlated (r2 = 0.938) to fracture load in bending. Load at failure ranged from 530 to 1,024 N (mean, 726 N; SD, 138 N), consistent with the findings of Miltner. Empirically derived strength parameters coupling BMD with geometry accurately predicted bending loads (r2 = 0.84, p < 0.001) and energy to failure (r2 = 0.88, p < 0.05). Torsional loading failed to generate spiral fractures consistently, resulting in either end plate or diaphyseal fractures. Load at failure for torsion ranged from 1,383 to 3,559 Newton-millimeters (mean, 2,703 Newton-millimeters; SD, 826 Newton-millimeters). Because of these inconsistent fracture results, empirical strength parameters for torsion could not be derived. CONCLUSION: BMD coupled with geometry is a strong predictor of bending fracture loads in the immature femoral diaphysis. A similar relationship could not be shown for torsion because of inconsistent failure results. This study represents an initial attempt at developing a methodology for predicting the strength of young bones from radiographic measures. Further research is required to establish this methodology and to show the necessary correlation with immature human bone.

Absorptiometry, Photon↗

The role of Yersinia enterocolitica and Yersinia pseudotuberculosis in granulomatous appendicitis: a histologic and molecular study.

Granulomatous appendicitis is an enigmatic entity. Purported causes include Crohn's disease, foreign body reactions, sarcoidosis, and infectious agents; however, most cases remain idiopathic. Yersinia enterocolitica (YE) and Y. pseudotuberculosis (YP) have been implicated as causes of appendicitis, ileocolitis, and mesenteric adenitis. The authors examined the potential role of YE and YP in granulomatous appendicitis using histologic and molecular methods. Forty cases of granulomatous appendicitis were evaluated for histologic features including transmural inflammation, number and character of granulomas, and mucosal changes. Twort Gram, Grocott methenamine-silver (GMS), and Ziehl-Neelsen stains were evaluated, and polymerase chain reaction (PCR) analysis was performed to identify pathogenic YP and YE. Twenty-five percent (10 of 40) of the cases were positive for pathogenic Yersinia by PCR (four YE, four YP, and two with both species). Prominent histologic features included epithelioid granulomas with lymphoid cuffing, transmural inflammation with lymphoid aggregates, mucosal ulceration, and cryptitis. One Yersinia-positive case contained mural Gram-negative bacilli; fungal and acid-fast bacilli stains were all negative. Except for one culture-negative case, serologies and cultures were not done or results were unavailable. Two Yersinia-positive patients were diagnosed subsequently with Crohn's disease, suggesting a possible relationship between the two entities. No other patients developed significant sequelae. YE and YP are important causes of granulomatous appendicitis, and Yersinia infection may mimic Crohn's disease. No histologic features distinguish reliably between Yersinia species, or between Yersinia-positive and Yersinia-negative cases. Because special stains and cultures are often not diagnostic, PCR analysis is an excellent technique for the diagnosis of Yersinia.

Adolescent↗

The role of smoking intentions in predicting future smoking among youth: findings from Monitoring the Future data.

AIMS: To demonstrate that intentions predict long-term future levels of smoking, irrespective of level of past smoking experience. A growing body of research suggests that intentions about future smoking might play an important role in addition to the influence of past smoking experience on the likelihood of smoking in future. DESIGN: Using logistic regression analyses, we assessed the relationship between baseline smoking experience and a firm intention 'not to be smoking cigarettes 5 years from now' with four outcome measures of smoking at follow-up: 30-day smoking at a 3/4- and 5/6-year follow-up and current established smoking (self-described regular smokers or former smokers who had smoked in the past 30 days) at a 3/4- and 5/6-year follow-up. PARTICIPANTS: US nationally representative samples of 12th graders who responded to the Monitoring the Future (MTF) survey from the years 1976 to 1993, inclusive. For these panels, we linked stage of smoking and intentions at 12th grade to follow-up measures of smoking collected at 3/4 years after baseline and 5/6 years after baseline. FINDINGS: Analysis of 3/4-year follow-up data (weighted n = 4544) and 5/6-year follow-up data (weighted n = 3885) for both definitions of smoking outcome indicated that there was a dose-response relationship between levels of baseline smoking experience and the likelihood of future smoking. In addition, independent of baseline smoking experience, there was a statistically significant protective effect for a firm intention not to smoke in five year's time on future smoking behavior. CONCLUSIONS: The findings suggest that evaluative studies of tobacco control policies and programs might usefully employ smoking uptake categories that incorporate smoking intentions as early indicators of outcome.

Adolescent↗

Genetic evidence for a SPO1-dependent signaling pathway controlling meiotic progression in yeast.

The yeast spindle pole body (SPB) plays a unique role in meiosis, initiating both spindle assembly and prospore membrane synthesis. SPO1, induced early in development, encodes a meiosis-specific phospholipase B (PLB) homolog required at three stages of SPB morphogenesis: MI, MII, and spore formation. Here we report in-depth analysis of the SPO1 gene including its transcriptional control by regulators of early gene expression, protein localization to the ER lumen and periplasmic space, and molecular genetic studies of its role in meiosis. Evidence is presented that multiple arrest points in spo1Delta occur independently, demonstrating that Spo1 acts at distinct steps. Loss of Spo1 is suppressed by high-copy glycosylphosphatidylinositol (GPI) proteins, dependent on sequence, timing, and strength of induction in meiosis. Since phosphatidylinositol (PI) serves as both an anchor component and a lipase substrate, we hypothesized that GPI-protein expression might substitute for Spo1 by decreasing levels of its potential substrates, PI and phosphatidylinositol phosphates (PIPs). Partial spo1Delta complementation by PLB3 (encoding a unique PLB capable of cleaving PI) and relatively strong Spo1 binding to PI(4)P derivatives (via a novel N-terminal lysine-rich fragment essential for Spo1 function) are consistent with this view. Epistasis of SPO1 mutations to those in SPO14 (encoding a PLD involved in signaling) and physical interaction of Spo1 with Spo23, a protein regulating PI synthesis required for wild-type sporulation, further support this notion. Taken together these findings implicate PI and/or PIPs in Spo1 function and suggest the existence of a novel Spo1-dependent meiosis-specific signaling pathway required for progression of MI, MII, and spore formation via regulation of the SPB.

Blotting, Southern↗

Evaluation of changes in trabecular bone architecture and mechanical properties of minipig vertebrae by three-dimensional magnetic resonance microimaging and finite element modeling.

The study objective was to analyze the three-dimensional (3D) trabecular architecture and mechanical properties in vertebral specimens of young and mature Sinclair minipigs to assess the relative contribution of architecture to bone strength. We used 3D magnetic resonance microimaging (MRmicroI) and direct image analysis to evaluate a set of standard structural measurements and new architectural descriptors of trabecular bone in biopsy specimens from L2, L3, and L4 vertebrae (n = 16 in each group) from young (mean age, 1.2 years) and mature (mean age, 4.8 years) minipigs. The measurements included bone volume/tissue volume (BV/TV), marrow star volume (Ma.St.V), connectivity density (ConnD), and two new parameters, percent platelike trabeculae (% plate) and percent bone in the load direction (% boneLD). The % plate, calculated from surface curvature, allowed the delineation of plates from rods. The % boneLD quantified the percentage of bone oriented along the long axis of the vertebral body. We showed that 3D MRmicroI can detect the subtle changes in trabecular architecture between the two age groups. ConnD, star volume, % plate, % boneLD, and BV/TV were found to be more effective than the model-based, derived indices (trabecular thickness [Tb.Th], trabecular separation [Tb.Sp], and trabecular number [Tb.N]) in differentiating the structural changes. BV/TV, % plate, and % boneLD significantly increased (p < 0.05) in all three vertebral sites of the mature minipigs. The significant decrease in ConnD and star volume in the mature vertebra was consistent with the concurrent increase of platelike trabecular bone (p < 0.05). Overall, ConnD, star volume, % plate, and % boneLD provided a coherent picture of the architectural changes between the two age groups. Apparent modulus and maximum stress were determined experimentally on biopsy specimens from L2 vertebrae (n = 16). When apparent modulus was predicted using 3D MRmicroI data sets as input for finite element modeling (FEM), the results were similar to the experimentally determined apparent modulus (p = 0.12). Both methods were then used to compare the young and the mature animals; the experimental and predicted apparent modulus were significantly higher for the mature group (p = 0.003 and 0.012, respectively). The experimental maximum stress in the vertebra of the mature animals was twice as high as that for the young animals (p = 0.006). Bone quantity (BV/TV or bone mineral content [BMC]) alone could explain approximately 74-85% of the total variability in stress and modulus. The inclusion of either ConnD or % boneLD with BV/TV in a multiple regression analysis significantly improved the predictability of maximum stress, indicating that architecture makes additional contributions to compressive strength in normal minipig vertebra.

Aging↗

Metabolic hormones modulate the effect of growth hormone (GH) on insulin-like growth factor-I (IGF-I) mRNA level in primary culture of salmon hepatocytes.

Liver production of insulin-like growth factor-I (IGF-I) is a major point of control in the growth hormone (GH)/IGF axis, the endocrine system regulating body growth in fishes and other vertebrates. Pituitary GH stimulates hepatocyte production of IGF-I; however, in catabolic states, hepatocyte GH resistance results in decreases in liver IGF-I production. To investigate endocrine mechanisms leading to the development of hepatocyte GH resistance, we examined the regulation of IGF-I mRNA level by GH and metabolic hormones in primary culture of salmon hepatocytes. Cells were cultured in RPMI medium, and exposed to insulin (Ins, 10(-6) M), glucagon (Glu, 10(-6) M), triiodothyronine (T3, 10(-7) M), dexamethasone (Dex, 10(-6) M) and glucagon-like peptide (GLP, 10(-6) M), in the presence and absence of GH (5 x 10(-9) M). GH always increased IGF-I mRNA. None of the other hormones tested alone affected IGF-I mRNA. However, Dex, Ins and Glu reduced the response to GH. The response to GH was inhibited by Dex at concentrations of 10(-12) M and above, by Ins at 10(-9) M and above, and by Glu only at 10(-6) M. Inhibition of GH response by glucocorticoids is found in other vertebrates. Salmon hepatocytes were very sensitive to Dex, suggesting that glucocorticoids may play an important role in salmon growth regulation even in unstressed conditions. Inhibition of GH response by Ins is the opposite of what is found in mammals and chickens, suggesting that the role of Ins in growth regulation may differ between fishes and tetrapods. To examine mechanisms for modulation of GH sensitivity, we measured hepatocyte GH receptor (GHR) mRNA levels. Ins inhibited and Dex stimulated GHR mRNA, suggesting that different mechanisms mediate the inhibition of GH response by these hormones. This study shows that glucocorticoids, Ins, and Glu induce GH resistance in cultured salmon hepatocytes.

Animals↗

Gemtuzumab ozogamicin with or without interleukin 11 in patients 65 years of age or older with untreated acute myeloid leukemia and high-risk myelodysplastic syndrome: comparison with idarubicin plus continuous-infusion, high-dose cytosine arabinoside.

We investigated treatment with gemtuzumab ozogamicin (GO) in 51 patients aged 65 years or older with newly diagnosed acute myeloid leukemia (AML), refectory anemia (RA) with excess of blasts in transformation, or RA with excess blasts. GO was given in doses of 9 mg/m(2) of body-surface area on days 1 and 8 or, therapeutically equivalently, on days 1 and 15, with or without interleukin 11 (IL-11; 15 microg/kg per day on days 3 to 28), with assignment to IL-11 treatment made randomly. Complete remission (CR) rates were 2 of 26 (8%) for GO without IL-11 and 9 of 25 (36%) for GO with IL-11. Regression analyses indicated that IL-11 was independently predictive of CR but not survival. We compared GO with or without IL-11 with idarubicin plus cytosine arabinoside (IA), as previously administered, in similar patients. The CR rate with IA was 15 of 31 (48%), and survival was superior with IA compared with GO with or without IL-11 (P =.03). Besides accounting for possible covariate effects on outcome, we also accounted for possible trial effects (TEs) arising because IA and GO with or without IL-11 were not arms of a randomized trial. Bayesian posterior probabilities that GO with or without IL-11 produced longer survival than IA, after accounting for covariates and TEs, were less than 0.01 in patients with abnormal cytogenetic findings (AC) and less than 0.15 in patients with normal cytogenetic findings (NC). Regarding CR, the analogous probabilities were less than 0.02 for GO without IL-11 (all cytogenetic groups), and for GO with IL-11, less than 0.25 for AC groups and about 0.50 for NC groups. TEs 2 to 5 times the magnitude of those previously observed would be needed to conclude that survival with GO with or without IL-11 is likely longer than with IA. Thus, there is little evidence to suggest that GO with or without IL-11 should be used instead of IA in older patients with newly diagnosed AML or myelodysplastic syndrome.

Acute Disease↗

Individual prior information in a physiological model of 2H8-toluene kinetics: an empirical Bayes estimation strategy.

Physiologically-based toxicokinetic (PBTK) models are widely used to quantify whole-body kinetics of various substances. However, since they attempt to reproduce anatomical structures and physiological events, they have a high number of parameters. Their identification from kinetic data alone is often impossible, and other information about the parameters is needed to render the model identifiable. The most commonly used approach consists of independently measuring, or taking fom literature sources, some of the parameters, fixing them in the kinetic model, and then performing model identification on a reduced number of less certain parameters. This results in a substantial reduction of the degrees of freedom of the model. In this study, we show that this method results in final estimates of the free parameters whose precision is overestimated. We then compared this approach with an empirical Bayes approach, which takes into account not only the mean value, but also the error associated with the independently determined parameters. Blood and breath 2H8-toluene washout curves, obtained in 17 subjects, were analyzed with a previously presented PBTK model suitable for person-specific dosimetry. Model parameters with the greatest effect on predicted levels were alveolar ventilation rate QPC, fat tissue fraction VFC, blood-air partition coefficient Kb, fraction of cardiac output to fat Qa/co and rate of extrahepatic metabolism Vmax-p. Differences in the measured and Bayesian-fitted values of QPC, VFC and Kb were significant (p < 0.05), and the precision of the fitted values Vmax-p and Qa/co went from 11 +/- 5% to 75 +/- 170% (NS) and from 8 +/- 2% to 9 +/- 2% (p < 0.05) respectively. The empirical Bayes approach did not result in less reliable parameter estimates: rather, it pointed out that the precision of parameter estimates can be overly optimistic when other parameters in the model, either directly measured or taken from literature sources, are treated as known without error. In conclusion, an empirical Bayes approach to parameter estimation resulted in a better model fit, different final parameter estimates, and more realistic parameter precisions.

Adult↗

Lectin binding profiles of SSEA-4 enriched, pluripotent human embryonic stem cell surfaces.

BACKGROUND: Pluripotent human embryonic stem cells (hESCs) have the potential to form every cell type in the body. These cells must be appropriately characterized prior to differentiation studies or when defining characteristics of the pluripotent state. Some developmentally regulated cell surface antigens identified by monoclonal antibodies in a variety of species and stem cell types have proven to be side chains of membrane glycolipids and glycoproteins. Therefore, to examine hESC surfaces for other potential pluripotent markers, we used a panel of 14 lectins, which were chosen based on their specificity for a variety of carbohydrates and carbohydrate linkages, along with stage specific embryonic antigen-4 (SSEA-4), to determine binding quantitation by flow cytometry and binding localization in adherent colonies by immunocytochemistry. RESULTS: Enriching cells for SSEA-4 expression increased the percentage of SSEA-4 positive cells to 98-99%. Using enriched high SSEA-4-expressing hESCs, we then analyzed the binding percentages of selected lectins and found a large variation in binding percentages ranging from 4% to 99% binding. Lycopersicon (tomato)esculetum lectin (TL), Ricinus communis agglutinin (RCA), and Concanavalin A (Con A) bound to SSEA-4 positive regions of hESCs and with similar binding percentages as SSEA-4. In contrast, we found Dolichos biflorus agglutinin (DBA) and Lotus tetragonolobus lectin (LTL) did not bind to hESCs while Phaseolus vulgaris leuco-agglutinin (PHA-L), Vicia villosa agglutinin (VVA), Ulex europaeus agglutinin (UEA), Phaseolus vulgaris erythro-agglutinin (PHA-E), and Maackia amurensis agglutinin (MAA) bound partially to hESCs. These binding percentages correlated well with immunocytochemistry results. CONCLUSION: Our results provide information about types of carbohydrates and carbohydrate linkages found on pluripotent hESC surfaces. We propose that TL, RCA and Con A may be used as markers that are associated with the pluripotent state of hESCs because binding percentages and binding localization of these lectins are similar to those of SSEA-4. Non-binding lectins, DBA and LTL, may identify differentiated cell types; however, we did not find these lectins to bind to pluripotent SSEA-4 positive hESCs. This work represents a fundamental base to systematically classify pluripotent hESCs, and in future studies these lectins may be used to distinguish differentiated hESC types based on glycan presentation that accompanies differentiation.

Antigens, Surface↗

Thoracoscopic discectomy and fusion in an animal model: safe and effective when segmental blood vessels are spared.

STUDY DESIGN: Disc-endplate excision and spine fusion were compared in animals randomly assigned to segmental vessel-spared and segmental vessel-ligated groups in an in vivo goat model of anterior spine discectomy and fusion using thoracoscopic techniques. OBJECTIVES: To compare safety and completeness of disc and endplate excision, and to perform a histologic and biomechanical comparison between fusion masses when the segmental vessels are spared and when they are ligated using thoracoscopic techniques. SUMMARY OFF BACKGROUND DATA: Because thoracoscopy is relatively new and technically demanding, many surgeons ligate the segmental blood vessels to enhance spine exposure and limit the risk of injury during discectomy and fusion. Although rare, spinal cord compromise secondary to segmental vessel ligation has been reported. METHODS: This study was divided into two phases. In Phase 1, 10 mature goats were randomly assigned to either the segmental vessel-ligated or the segmental vessel-spared group. Disc and endplate excision was performed at six consecutive thoracic levels in each animal (30 levels per group). The animals were killed, and the depth of disc excision was measured in the transverse and sagittal planes. The vertebral bodies then were separated through the disc space; photographic images of the endplates were digitized, and the area of endplate excision was calculated. In Phase 2, 12 mature goats were randomly assigned to the segmental vessel-ligated or vessel-spared group, and five noncontiguous thoracic segments were fused using autologous iliac crest graft. At 4 months the animals were killed, and the spines were harvested. At each disc level, the three-dimensional rotational and translational motions were analyzed and histomorphometric analysis was performed. RESULTS: Phase 1: Each animal survived the operative procedure, and no surgical complications occurred. No difference was found between vessel-ligated and vessel-spared groups in operative time (21.8 vs 22.7 minutes per disc), blood loss (97 vs 159 mL), or transverse (81% vs 74%) or sagittal (85% vs 85%) disc excision. The total area of endplate excision was 70% in the vessel-ligated group and 67% in the vessel-spared group (P > 0.1). Phase 2: Biomechanical testing demonstrated no difference in stiffness of the fused segments between the two groups in flexion-extension or axial rotation. However, greater flexibility in lateral bending was found in the specimens whose vessels were ligated (P < 0.05). The percentage of trabecular bone volume was similar between the two groups. CONCLUSIONS: The segmental vessels in the thoracic spine can be effectively spared without injury during disc excision and fusion. Although slightly more disc area was excised with ligation of the vessels, this was not statistically significant, and the fusion mass was similar between the two groups. Sparing the segmental vessels may provide blood supply that aids fusion mass, and the result may be greater spine stiffness in the coronal plane. Sparing the segmental vessels during thoracoscopic anterior disc excision and fusion can be safe. It should be considered in patients with a high risk for neurologic injury because of decreased spinal cord perfusion in revision surgery, severe kyphosis, congenital anomalies. Because the neurologic risk of vessel ligation has not been clearly established for idiopathic scoliosis, the surgeon will have to consider the risk-benefit ratio of adopting these methods when deciding not to ligate vessels in these patients.

Animals↗