Soy components vs. whole soy: are we betting our bones on a long shot? Soybeans are a good source of bone-healthy nutrients. Epidemiological studies in Asia evaluating diets containing traditional whole soy foods show a positive association with bone mineral density and fracture protection. Smaller scale intervention studies in Western nations mainly feature isolated soy protein (SP) and purified or concentrated soy isoflavones (SI) rather than whole soy foods and they have produced inconsistent results. Consumption of SP does not alter calcium (Ca) retention even though urinary Ca excretion is less in diets with SP compared with proteins higher in sulfur-containing amino acids. SI, often consumed at higher concentrations than would be available in traditional Asian diets, are not yielding the type of incontrovertible evidence that might be expected in support of their benefit to bone health. This forces one to ask whether whole soy might provide a superior effect on bone.


A ready-to-use therapeutic food containing 10% milk is less effective than one with 25% milk in the treatment of severely malnourished children. Standard therapy for severe acute malnutrition (SAM) is home-based therapy with ready-to-use therapeutic food (RUTF) containing 25% milk. In an effort to lower the cost of RUTF and increase availability, some have suggested that a portion of milk be replaced with soy. This trial was designed to determine whether treating children with SAM with 10% milk RUTF containing soy would result in a similar recovery rate compared with the 25% milk RUTF. This was a randomized, double-blind, controlled, clinical, quasi-effectiveness trial of isoenergetic amounts of 2 locally produced RUTF to treat SAM in Malawi among children aged 6-59 mo. A total of 1874 children were enrolled. Children were assessed every fortnight and participated in the study until they clinically recovered or received 8 wk of treatment. The primary outcome was recovery (weight-for-height Z score > -2 and no edema). Secondary outcomes were rates of weight and height gain. Survival analysis was used to compare the recovery rates. Recovery among children receiving 25% milk RUTF was greater than children receiving 10% milk RUTF, 64% compared with 57% after 4 wk, and 84% compared with 81% after 8 wk (P < 0.001). Children receiving 25% milk RUTF also had higher rates of weight and height gain compared with children receiving 10% milk RUTF. Treating children with SAM with 10% milk RUTF is less effective compared with treatment with the standard 25% milk RUTF. These findings also emphasize that clinical evidence should be examined before recommending any changes to the formulation of RUTF.


Beta-carotene is an important vitamin A source for humans. Experts in the field of carotenoids met at the Hohenheim consensus conference in July 2009 to elucidate the current status of β-carotene research and to summarize the current knowledge with respect to the chemical properties, physiological function, and intake of β-carotene. The experts discussed 17 questions and reached an agreement formulated in a consensus answer in each case. These consensus answers are based on published valid data, which were carefully reviewed by the individual experts and are justified here by background statements. Ascertaining the impact of β-carotene on the total dietary intake of vitamin A is complicated, because the efficiency of conversion of β-carotene to retinol is not a single ratio and different conversion factors have been used in various surveys and following governmental recommendations within different countries. However, a role of β-carotene in fulfilling the recommended intake for vitamin A is apparent from a variety of studies. Thus, besides elucidating the various functions, distribution, and uptake of β-carotene, the consensus conference placed special emphasis on the provitamin A function of β-carotene and the role of β-carotene in the realization of the required/recommended total vitamin A intake in both developed and developing countries. There was consensus that β-carotene is a safe source of vitamin A and that the provitamin A function of β-carotene contributes to vitamin A intake.


Rye whole grain and bran intake compared with refined wheat decreases urinary C-peptide, plasma insulin, and prostate specific antigen in men with prostate cancer. Rye whole grain and bran intake has shown beneficial effects on prostate cancer progression in animal models, including lower tumor take rates, smaller tumor volumes, and reduced prostate specific antigen (PSA) concentrations. A human pilot study showed increased apoptosis after consumption of rye bran bread. In this study, we investigated the effect of high intake of rye whole grain and bran on prostate cancer progression as assessed by PSA concentration in men diagnosed with prostate cancer. Seventeen participants were provided with 485 g rye whole grain and bran products (RP) or refined wheat products with added cellulose (WP), corresponding to ~50% of daily energy intake, in a randomized controlled, crossover design. Blood samples were taken from fasting men before and after 2, 4, and 6 wk of treatment and 24-h urine samples were collected before the first intervention period and after treatment. Plasma total PSA concentrations were lower after treatment with RP compared with WP, with a mean treatment effect of -14% (P = 0.04). Additionally, fasting plasma insulin and 24-h urinary C-peptide excretion were lower after treatment with RP compared with WP (P < 0.01 and P = 0.01, respectively). Daily excretion of 5 lignans was higher after the RP treatment than after the WP treatment (P < 0.001). We conclude that whole grain and bran from rye resulted in significantly lower plasma PSA compared with a cellulose-supplemented refined wheat diet in patients with prostate cancer. The effect may be related to inhibition of prostate cancer progression caused by decreased exposure to insulin, as indicated by plasma insulin and urinary C-peptide excretion.


A low-protein diet during gestation in rats activates the placental mammalian amino acid response pathway and programs the growth capacity of offspring. Placental efficiency is a predictor of fetal growth and development, which is also controlled by maternal gestational health and diet. The present study investigated the effects of a gestational low-protein diet on offspring growth capacity as well as the diet's contribution to altered expression of placental genes associated with the mammalian amino acid response (AAR) pathway. To assess these outcomes, timed-pregnant Sprague Dawley rats were fed a control (C) diet with 18% protein or a low-protein (LP) diet with 9% protein throughout gestation (Expt. 1) or throughout gestation and lactation (Expt. 2). Placentas were collected during natural delivery and quantitative RT-PCR and Western-blot analyses were performed to determine placental mRNA and protein levels. By the end of the lactation period, offspring of dams fed the LP diet had stunted growth in both experiments. mRNA expression of target genes in the AAR pathway, such as activating transcription factor-3 (Atf3), asparagine synthetase (Asns), and Sodium-dependent neutral amino acid transporter-2 (Snat2), was greater in placentas of rats fed the LP diet compared with controls, as were placental ATF4 and p-eIF2α protein levels. The increase in mRNA expression of AAR pathway-associated genes was correlated with the stunting of offspring growth (Atf3: R(2) = 0.32, P = 0.086; Asns: R(2) = 0.44, P < 0.05; Snat2: R(2) = 0.33, P = 0.084). Our study showed that the mammalian AAR pathway in placenta is upregulated by a maternal low-protein diet and this activation may act as a cue for the fetus to develop an adaptive response suited to their predicted postnatal environment, i.e. a more favorable phenotype for their survival.


Whole versus the piecemeal approach to evaluating soy. Soy has been singled out for attention among other legumes as a valuable source of nutrients, phytochemicals, and bioactive compounds. Early epidemiological studies established that whole soy and traditional soy foods were implicated in health-protective effects in Asian populations. The same benefits attributable to soy have not been consistently proven in Western populations that, for various reasons, opt to consume more processed soy foods or various soy components. Soy researchers continue to isolate soy components in search of identifying its salubrious components and whole soy remains relatively underinvestigated despite what we know of the health benefits it may confer to those regularly consuming it. Various dietary guidelines advocate the regular consumption of legumes that tend not to be included in our diets in sufficient quantities. This paper highlights the possibility that whole soy may have a more unique effect on health than a select soy component(s). It explores the rationale for focusing research on whole soy in an attempt to understand it better rather than trying to replicate the health benefits by targeting various soy components, which has been plagued by inconsistent results.


How much human milk do infants consume? Data from 12 countries using a standardized stable isotope methodology. The WHO has developed new growth curves based on breast-fed infants. Recommendations for energy intake have been adopted based on measurements of total energy expenditure. Data on human milk (HM) intake are needed to estimate the energy intake from this food source. However, objective HM data from around the world have not been available, because these measurements are difficult to obtain. Stable isotope methods have been developed to provide objective measurements over a 14-d period. A pooled analysis of 1115 data points of HM intake, obtained using the dose to the mother deuterium oxide turnover method, was undertaken in infants aged 0-24 mo from 12 countries across 5 continents. A hierarchical model was needed to estimate mean HM intake and its variance within and between countries given the complexity of the data. The overall mean HM intake was 0.78 (95% CI = 0.72, 0.84) kg/d, and the age-specific estimates indicated that intake increased over the first 3-4 mo and remained above 0.80 kg/d until 6-7 mo. The variability of intake increased in late infancy. Boys consumed 0.05 kg/d more than girls (P < 0.01). HM intake was strongly, inversely associated with non-HM water intake [r = -0.448 (95% CI -0.511 to -0.385); P < 0.0001]. These objective isotope values of HM intake improve our understanding of the magnitude and variability of HM intake within and across populations and help to estimate nutrient intakes in breast-fed infants.


Serum 25-hydroxyvitamin D concentrations fluctuate seasonally in young adults of diverse ancestry living in Toronto. Previous research indicates that circulating vitamin D levels are low in many otherwise healthy adults and that there is considerable seasonal variation in 25-hydroxyvitamin D [25(OH)D] concentrations at high latitudes. We examined seasonal variation in 25(OH)D levels in a sample of young adults of diverse ancestry living in the Greater Toronto Area. Three hundred and fifty-one (351) healthy young adults completed both a fall and winter visit during this study. The study was conducted over 2 y (y 1: fall 2007 to winter 2008 and y 2: fall 2008 to winter 2009). At both visits, each participant's serum 25(OH)D concentration was measured. Information was also obtained on skin pigmentation (measured via reflectometer), vitamin D intake, and extent of sun exposure. Overall, the serum 25(OH)D concentration was 54.4 ± 1.3 nmol/L in the fall and 38.4 ± 1.1 nmol/L in the winter. Concentrations differed among ancestral groups at both visits (P < 0.001), with South Asians and East Asians having substantially lower concentrations than Europeans. Skin pigmentation (r(2) = 0.14; P < 0.001), supplemental vitamin D intake (r(2) = 0.09; P < 0.001), sun exposure (r(2) = 0.04; P < 0.001), and study year (r(2) = 0.02; P = 0.017) were predictors of fall 25(OH)D concentrations. During the wintertime, serum 25(OH)D concentrations were associated with concentrations taken in the fall (r(2) = 0.45; P < 0.001), supplemental (r(2) = 0.15; P < 0.001) and dietary vitamin D intake (r(2) = 0.06; P < 0.001), and with study year (r(2) = 0.02; P = 0.009). Our study confirms that serum 25(OH)D concentrations undergo strong seasonal variation at high latitudes and are influenced by vitamin D intake, skin pigmentation, and sun exposure.


Consumption of wheat aleurone-rich foods increases fasting plasma betaine and modestly decreases fasting homocysteine and LDL-cholesterol in adults. There is strong evidence that whole-grain foods protect against heart disease. Although underlying mechanisms and components are unclear, betaine, found at high levels in wheat aleurone, may play a role. We evaluated the effects of a diet high in wheat aleurone on plasma betaine and related measures. In a parallel, single-blinded intervention study, 79 healthy participants (aged 45-65 y, BMI ≥ 25 kg/m(2)) incorporated either aleurone-rich cereal products (27 g/d aleurone) or control products balanced for fiber and macronutrients into their habitual diets for 4 wk. Fasting blood samples were taken at baseline and postintervention (4 wk) from participants. Compared with the control, the aleurone products provided an additional 279 mg/d betaine and resulted in higher plasma betaine (P < 0.001; intervention effect size: 5.2 μmol/L) and lower plasma total homocysteine (tHcy) (P = 0.010; -0.7 μmol/L). Plasma dimethylglycine and methionine, which are products of betaine-mediated homocysteine remethylation, were also higher (P < 0.001; P = 0.027) relative to control. There were no significant effects on plasma choline or B vitamins (folate, riboflavin, and vitamin B-6). However, LDL cholesterol was lower than in the control group (P = 0.037). We conclude that incorporating aleurone-rich products into the habitual diet for 4 wk significantly increases plasma betaine concentrations and lowers tHcy, which is attributable to enhanced betaine-homocysteine methyltransferase-mediated remethylation of homocysteine. Although this supports a role for betaine in the protective effects of whole grains, concomitant decreases in LDL suggest more than one component or mechanism may be responsible.


Preface to journal supplement "Soy Summit: Exploration of the Nutrition and Health Effects of Whole Soy".


Web-enabled and improved software tools and data are needed to measure nutrient intakes and physical activity for personalized health research. Food intake, physical activity (PA), and genetic makeup each affect health and each factor influences the impact of the other 2 factors. Nutrigenomics describes interactions between genes and environment. Knowledge about the interplay between environment and genetics would be improved if experimental designs included measures of nutrient intake and PA. Lack of familiarity about how to analyze environmental variables and ease of access to tools and measurement instruments are 2 deterrents to these combined studies. This article describes the state of the art for measuring food intake and PA to encourage researchers to make their tools better known and more available to workers in other fields. Information presented was discussed during a workshop on this topic sponsored by the USDA, NIH, and FDA in the spring of 2009.


Complementary foods fortified with micronutrients prevent iron deficiency and anemia in Vietnamese infants. To investigate whether an intervention including micronutrient-fortified complementary foods can improve iron status, we conducted a randomized controlled trial in 5-mo-old Vietnamese infants (n = 246). Villages (n = 29) were randomly divided into those receiving instant flour (FF) or a food complement (FC) both fortified with micronutrients or nothing [control (C)]. FF and FC infants received daily for 6 mo at least 2 meals of fortified complementary foods. Micronutrient status was assessed by measurement of hemoglobin (Hb) and plasma ferritin (PF), transferrin receptor, zinc, and retinol. Final Hb (mean ± SD) was higher in the FF (112.5 ± 8.0 g/L) and FC (114.0 ± 7.0 g/L) groups compared with C (109.0 ± 8.0 g/L; P = 0.006). PF (geometric mean [95% CI]) was also higher in FF (19.8 μg/L [17.5-22.3]) and FC (20.8 μg/L [18.3-23.6]) compared with C (11.1 μg/L [9.8-12.5]; P < 0.0001). Anemia prevalence decreased more in the FC group (-43.6%) compared with C (-10.3%; P = 0.006). The change in prevalence of PF < 12 μg/L was different in the FF (-16.4%) and FC (-6.7%) groups compared with C (+30.4%; P < 0.01). Endpoint prevalence of iron deficiency (ID) and ID anemia (IDA) were lower in the FF (13.4 and 6.7%, respectively) and FC (15.2 and 3.8%) groups compared with C (57.5 and 37.5%) (P < 0.0001). Retinol and zinc concentrations did not differ among groups, but endpoint prevalence of zinc deficiency was lower in FF infants (36.1%) than in C infants (52.9%; P = 0.04). Micronutrient-fortified complementary foods significantly improved iron status and decreased the prevalence of anemia, ID, and IDA in Vietnamese infants and can be an important tool to reduce ID in infancy in developing countries.


Utilization of iron from an animal-based iron source is greater than that of ferrous sulfate in pregnant and nonpregnant women. Heme iron absorption during pregnancy and the role of hepcidin in regulating dietary heme iron absorption remains largely unexplored. The objective of this research was to examine relative differences in heme (animal based) and nonheme (ferrous sulfate) iron utilization. This study was undertaken in 18 pregnant (ages 16-32 y; wk 32-35 of gestation) and 11 nonpregnant women (ages 18-27 y). Women were randomly assigned to receive both an animal-based heme meal (intrinsically labeled (58)Fe pork) and labeled ferrous sulfate ((57)Fe) fed on alternate days. Blood samples obtained 2 wk postdosing were used to assess iron status indicators and serum hepcidin and iron utilization based on RBC incorporation of iron isotopes. Heme iron utilization was significantly greater than nonheme iron utilization in the pregnant (47.7 ± 14.4 vs. 40.4 ± 13.2%) and nonpregnant women (50.1 ± 14.8 vs. 15.3 ± 9.7%). Among pregnant women, utilization of nonheme iron was associated with iron status, as assessed by the serum transferrin receptor concentration (P = 0.003; r(2) = 0.43). In contrast, heme iron utilization was not influenced by maternal iron status. In the group as a whole, women with undetectable serum hepcidin had greater nonheme iron utilization compared with women with detectable serum hepcidin (P = 0.02; n = 29); however, there were no significant differences in heme iron utilization. Our study suggests that iron utilization from an animal-based food provides a highly bioavailable source of dietary iron for pregnant and nonpregnant women that is not as sensitive to hepcidin concentrations or iron stores compared with ferrous sulfate.


A DXA-based mathematical model predicts midthigh muscle mass from magnetic resonance imaging in typically developing children but not in those with quadriplegic cerebral palsy. Valid methods for assessing regional muscle mass in children are needed. The aim of this study was to determine whether dual-energy X-ray absorptiometry (DXA) can accurately estimate midthigh muscle mass from MRI (muscle(MRI)) in typically developing children and children with quadriplegic cerebral palsy (CP). A mathematical model predicting muscle(MRI) from midthigh, fat-free soft tissue mass from DXA (FFST(DXA)) was developed using 48 typically developing children (6-13 y) and was validated using the leave-one-out method. The model was also tested in children with quadriplegic CP (n = 10). The model produced valid estimates of midthigh muscle mass (muscle(DXA)) in typically developing children, as indicated by a very strong relationship between muscle(DXA) and muscle(MRI) (r(2) = 0.95; SEE = 68 g; P < 0.001), no difference in muscle(DXA) and muscle(MRI) (P = 0.951), and visual examination using a Bland-Altman plot. Muscle(DXA) was very strongly related to muscle(MRI) in children with CP (r(2) = 0.96; SEE = 54 g; P < 0.001); however, muscle(DXA) overestimated muscle(MRI) by 15% (P = 0.006). The overestimation of muscle(MRI) by muscle(DXA) was strongly related to the lower ratio of muscle(MRI) to FFST(DXA) (muscle(MRI)/FFST(DXA)) in children with CP (r(2) = 0.75; P = 0.001). The findings suggest that the DXA-based mathematical model developed in the current study can accurately estimate midthigh muscle mass in typically developing children. However, a population-specific model that takes into account the lower muscle(MRI)/FFST(DXA) is needed to estimate midthigh muscle mass in children with quadriplegic CP.


Cryo-EM structure and rRNA model of a translating eukaryotic 80S ribosome at 5.5-A resolution. Protein biosynthesis, the translation of the genetic code into polypeptides, occurs on ribonucleoprotein particles called ribosomes. Although X-ray structures of bacterial ribosomes are available, high-resolution structures of eukaryotic 80S ribosomes are lacking. Using cryoelectron microscopy and single-particle reconstruction, we have determined the structure of a translating plant (Triticum aestivum) 80S ribosome at 5.5-Å resolution. This map, together with a 6.1-Å map of a Saccharomyces cerevisiae 80S ribosome, has enabled us to model ∼98% of the rRNA. Accurate assignment of the rRNA expansion segments (ES) and variable regions has revealed unique ES-ES and r-protein-ES interactions, providing insight into the structure and evolution of the eukaryotic ribosome.


Inositol hexakisphosphate kinase-2 acts as an effector of the vertebrate Hedgehog pathway. Inositol phosphate (IP) kinases constitute an emerging class of cellular kinases linked to multiple cellular activities. Here, we report a previously uncharacterized cellular function in Hedgehog (Hh) signaling for the IP kinase designated inositol hexakisphosphate kinase-2 (IP6K2) that produces diphosphoryl inositol phosphates (PP-IPs). In zebrafish embryos, IP6K2 activity was required for normal development of craniofacial structures, somites, and neural crest cells. ip6k2 depletion in both zebrafish and mammalian cells also inhibited Hh target gene expression. Inhibiting IP(6) kinase activity using N(2)-(m-(trifluoromethy)lbenzyl) N(6)-(p-nitrobenzyl)purine (TNP) resulted in altered Hh signal transduction. In zebrafish, restoring IP6K2 levels with exogenous ip6k2 mRNA reversed the effects of IP6K2 depletion. Furthermore, overexpression of ip6k2 in mammalian cells enhanced the Hh pathway response, suggesting IP6K2 is a positive regulator of Hh signaling. Perturbations from IP6K2 depletion or TNP were reversed by overexpressing smoM2, gli1, or ip6k2. Moreover, the inhibitory effect of cyclopamine was reversed by overexpressing ip6k2. This identified roles for the inositol kinase pathway in early vertebrate development and tissue morphogenesis, and in Hh signaling. We propose that IP6K2 activity is required at the level or downstream of Smoothened but upstream of the transcription activator Gli1.


Pineal-specific agouti protein regulates teleost background adaptation. Background adaptation is used by teleosts as one of a variety of camouflage mechanisms for avoidance of predation. Background adaptation is known to involve light sensing by the retina and subsequent regulation of melanophore dispersion or contraction in melanocytes, mediated by α-melanocyte-stimulating hormone and melanin-concentrating hormone, respectively. Here, we demonstrate that an agouti gene unique to teleosts, agrp2, is specifically expressed in the pineal and is required for up-regulation of hypothalamic pmch and pmchl mRNA and melanosome contraction in dermal melanocytes in response to a white background. floating head, a mutant with defective pineal development, exhibits defective up-regulation of mch mRNAs by white background, whereas nrc, a blind mutant, exhibits a normal response. These studies identify a role for the pineal in background adaptation in teleosts, a unique physiological function for the agouti family of proteins, and define a neuroendocrine axis by which environmental background regulates pigmentation.


A network connecting Runx2, SATB2, and the miR-23a~27a~24-2 cluster regulates the osteoblast differentiation program. Induced osteogenesis includes a program of microRNAs (miRs) to repress the translation of genes that act as inhibitors of bone formation. How expression of bone-related miRs is regulated remains a compelling question. Here we report that Runx2, a transcription factor essential for osteoblastogenesis, negatively regulates expression of the miR cluster 23a∼27a∼24-2. Overexpression, reporter, and chromatin immunoprecipitation assays established the presence of a functional Runx binding element that represses expression of these miRs. Consistent with this finding, exogenous expression of each of the miRs suppressed osteoblast differentiation, whereas antagomirs increased bone marker expression. The biological significance of Runx2 repression of this miR cluster is that each miR directly targets the 3' UTR of SATB2, which is known to synergize with Runx2 to facilitate bone formation. The findings suggest Runx2-negative regulation of multiple miRs by a feed-forward mechanism to cause derepression of SATB2 to promote differentiation. We find also that miR-23a represses Runx2 in the terminally differentiated osteocyte, representing a feedback mechanism to attenuate osteoblast maturation. We provide direct evidence for an interdependent relationship among transcriptional inhibition of the miR cluster by Runx2, translational repression of Runx2 and of SATB2 by the cluster miRs during progression of osteoblast differentiation. Furthermore, miR cluster gain of function (i.e., inhibition of osteogenesis) is rescued by the exogenous expression of SATB2. Taken together, we have established a regulatory network with a central role for the miR cluster 23a∼27a∼24-2 in both progression and maintenance of the osteocyte phenotype.


Informatics in radiology: an information model of the DICOM standard. The Digital Imaging and Communications in Medicine (DICOM) Standard is a key foundational technology for radiology. However, its complexity creates challenges for information system developers because the current DICOM specification requires human interpretation and is subject to nonstandard implementation. To address this problem, a formally sound and computationally accessible information model of the DICOM Standard was created. The DICOM Standard was modeled as an ontology, a machine-accessible and human-interpretable representation that may be viewed and manipulated by information-modeling tools. The DICOM Ontology includes a real-world model and a DICOM entity model. The real-world model describes patients, studies, images, and other features of medical imaging. The DICOM entity model describes connections between real-world entities and the classes that model the corresponding DICOM information entities. The DICOM Ontology was created to support the Cancer Biomedical Informatics Grid (caBIG) initiative, and it may be extended to encompass the entire DICOM Standard and serve as a foundation of medical imaging systems for research and patient care.


APOE varepsilon2 and presymptomatic stage Alzheimer disease: how much is not enough?


A network approach to assessing cognition in disorders of consciousness. Conventional assessments of consciousness rely on motor responses to indicate awareness. However, overt behaviors may be absent or ambiguous in patients with disorders of consciousness (DOC) resulting in underrating capacity for cognition. fMRI during a silent picture-naming task was evaluated as an indicator of command following when conventional methods are not sufficient. A total of 10 patients with and without conventional evidence of awareness, who met diagnostic criteria for the minimally conscious state (MCS) (n = 5), vegetative state (VS) (n = 3), emerged from MCS (EMCS) (n = 1), and locked-in syndrome (LIS) (n = 1), participated in this observational fMRI study. The LIS and EMCS patients engaged a complete network of essential language-related regions during the object-naming task. The MCS and 2 of the VS patients demonstrated both complete and partial preservation of the object-naming system. Patients who engaged a complete network scored highest on the Coma Recovery Scale-Revised. This study supports the view that fMRI during object naming can elicit brain activations in patients with DOC similar to those observed in healthy subjects during command following, and patients can be stratified by completeness of the engaged neural system. These results suggest that activity of the language network may serve as an indicator of high-level cognition and possibly volitional processes that cannot be discerned through conventional behavioral assessment alone.


Application of failure mode and effect analysis in a radiology department. With increasing deployment, complexity, and sophistication of equipment and related processes within the clinical imaging environment, system failures are more likely to occur. These failures may have varying effects on the patient, ranging from no harm to devastating harm. Failure mode and effect analysis (FMEA) is a tool that permits the proactive identification of possible failures in complex processes and provides a basis for continuous improvement. This overview of the basic principles and methodology of FMEA provides an explanation of how FMEA can be applied to clinical operations in a radiology department to reduce, predict, or prevent errors. The six sequential steps in the FMEA process are explained, and clinical magnetic resonance imaging services are used as an example for which FMEA is particularly applicable. A modified version of traditional FMEA called Healthcare Failure Mode and Effect Analysis, which was introduced by the U.S. Department of Veterans Affairs National Center for Patient Safety, is briefly reviewed. In conclusion, FMEA is an effective and reliable method to proactively examine complex processes in the radiology department. FMEA can be used to highlight the high-risk subprocesses and allows these to be targeted to minimize the future occurrence of failures, thus improving patient safety and streamlining the efficiency of the radiology department.


Can functional imaging assess consciousness in noncommunicative patients?


Hippocampal atrophy rates and CSF biomarkers in elderly APOE2 normal subjects. To determine whether elderly normal APOE E2 (APOE2) carriers exhibit slower rates of hippocampal atrophy and memory decline compared to APOE3/3 carriers. We also determined whether APOE2 carriers have less Alzheimer pathology as reflected by CSF biomarkers. We included longitudinal data from 134 cognitively normal individuals (27 APOE2/2 or E2/3, 107 APOE3/3) from the Alzheimer's Disease Neuroimaging Initiative, a prospective cohort study. A linear mixed-effects model was used to determine how APOE2 affected rates of hippocampal atrophy and cognitive change over time. In a subsample of 72 individuals who also underwent CSF analysis, an ordinary least-squares regression was used to determine whether CSF β-amyloid (Aβ), total tau, and phosphorylated tau-181 (p-tau) differed by APOE2 status. APOE2 carriers demonstrated slower rates of hippocampal atrophy (p = 0.004). The mean rate of hippocampal atrophy among APOE2 carriers was -33 mm(3)/year (95% confidence interval -65 to +0.4), or -0.5%/year, compared to -86 mm(3)/year (95% confidence interval -102 to -71), or -1.3%/year, in the APOE3/3 group. No differences in the rates of episodic memory (p = 0.23) or overall cognitive change (p = 0.90) were detected. In the CSF subsample, APOE2 carriers had higher levels of CSF Aβ (p = 0.01), lower p-tau (p = 0.02), and marginally lower tau (p = 0.12). A slower rate of hippocampal atrophy in normal APOE2 carriers is consistent with the lower risk of Alzheimer disease in these individuals. We hypothesize that the slower atrophy rate is related to decreased preclinical Alzheimer pathology.


Mechanism of N-methylation by the tRNA m1G37 methyltransferase Trm5. Trm5 is a eukaryal and archaeal tRNA methyltransferase that catalyzes methyl transfer from S-adenosylmethionine (AdoMet) to the N(1) position of G37 directly 3' to the anticodon. While the biological role of m(1)G37 in enhancing translational fidelity is well established, the catalytic mechanism of Trm5 has remained obscure. To address the mechanism of Trm5 and more broadly the mechanism of N-methylation to nucleobases, we examined the pH-activity profile of an archaeal Trm5 enzyme, and performed structure-guided mutational analysis. The data reveal a marked dependence of enzyme-catalyzed methyl transfer on hydrogen ion equilibria: the single-turnover rate constant for methylation increases by one order of magnitude from pH 6.0 to reach a plateau at pH 7.0. This suggests a mechanism involving proton transfer from G37 as the key element in catalysis. Consideration of the kinetic data in light of the Trm5-tRNA-AdoMet ternary cocrystal structure, determined in a precatalytic conformation, suggests that proton transfer is associated with an induced fit rearrangement of the complex that precedes formation of the reactive configuration in the active site. Key roles for the conserved R145 side chain in stabilizing a proposed oxyanion at G37-O(6), and for E185 as a general base to accept the proton from G37-N(1), are suggested based on the mutational analysis.


Characterization of purified human Bact spliceosomal complexes reveals compositional and morphological changes during spliceosome activation and first step catalysis. To better understand the compositional and structural dynamics of the human spliceosome during its activation, we set out to isolate spliceosomal complexes formed after precatalytic B but prior to catalytically active C complexes. By shortening the polypyrimidine tract of the PM5 pre-mRNA, which lacks a 3' splice site and 3' exon, we stalled spliceosome assembly at the activation stage. We subsequently affinity purified human B(act) complexes under the same conditions previously used to isolate B and C complexes, and analyzed their protein composition by mass spectrometry. A comparison of the protein composition of these complexes allowed a fine dissection of compositional changes during the B to B(act) and B(act) to C transitions, and comparisons with the Saccharomyces cerevisiae B(act) complex revealed that the compositional dynamics of the spliceosome during activation are largely conserved between lower and higher eukaryotes. Human SF3b155 and CDC5L were shown to be phosphorylated specifically during the B to B(act) and B(act) to C transition, respectively, suggesting these modifications function at these stages of splicing. The two-dimensional structure of the human B(act) complex was determined by electron microscopy, and a comparison with the B complex revealed that the morphology of the human spliceosome changes significantly during its activation. The overall architecture of the human and S. cerevisiae B(act) complex is similar, suggesting that many of the higher order interactions among spliceosomal components, as well as their dynamics, are also largely conserved.


Conformational distributions at the N-peptide/boxB RNA interface studied using site-directed spin labeling. In bacteriophage λ, interactions between a 22-amino acid peptide (called the N-peptide) and a stem-loop RNA element (called boxB) play a critical role in transcription anti-termination. The N-peptide/boxB complex has been extensively studied, and serves as a paradigm for understanding mechanisms of protein/RNA recognition. Particularly, ultrafast spectroscopy techniques have been applied to monitor picosecond fluorescence decay behaviors of 2-aminopurines embedded at various positions of the boxB RNA. The studies have led to a model in which the bound N-peptide exists in dynamic equilibrium between two states, with peptide C-terminal fragment either stacking on (i.e., the stacked state) or peeling away from (i.e., the unstacked state) the RNA loop. The function of the N-peptide/boxB complex seems to correlate with the fraction of the stacked state. Here, the N-peptide/boxB system is studied using the site-directed spin labeling technique, in which X-band electron paramagnetic resonance spectroscopy is applied to monitor nanosecond rotational behaviors of stable nitroxide radicals covalently attached to different positions of the N-peptide. The data reveal that in the nanosecond regime the C-terminal fragment of bound N-peptide adopts multiple discrete conformations within the complex. The characteristics of these conformations are consistent with the proposed stacked and unstacked states, and their distributions vary upon mutations within the N-peptide. These results suggest that the dynamic two-state model remains valid in the nanosecond regime, and represents a unique mode of function in the N-peptide/boxB RNA complex. It also demonstrates a connection between picosecond and nanosecond dynamics in a biological complex.


Biogenesis pathways of piRNAs loaded onto AGO3 in the Drosophila testis. PIWI-interacting RNAs (piRNAs) silence transposable elements in animal germ cells. In Drosophila ovaries, piRNAs are produced by two distinct pathways: the "ping-pong" amplification cycle that operates in germ cells and a ping-pong-independent pathway termed the primary pathway that mainly operates in somatic cells. AGO3, one of three PIWI proteins in flies, is involved in the ping-pong cycle in ovaries. We characterized AGO3-associated piRNAs in fly testes and found that like in ovaries, AGO3 functions in the ping-pong cycle with Aubergine (Aub) for piRNA production from transposon transcripts. In contrast, most AGO3-associated piRNAs corresponding to Suppressor of Stellate [Su(Ste)] genes are antisense-oriented and bound to Aub. In addition, the vast majority of AGO3-bound piRNAs derived from the AT-chX locus on chromosome X are antisense-oriented and are also found among Aub-associated piRNAs. The presence of very few sense Su(Ste) and AT-chX piRNAs suggests that biogenesis of both Su(Ste) and AT-chX piRNAs by a ping-pong mechanism only is highly unlikely. Nevertheless, the mutual interdependence of AGO3 and Aub for the accumulation of these piRNAs shows that their production relies on both AGO3 and Aub. Analysis of piRNA pathway mutants revealed that although the requirements for piRNA factors for Su(Ste)- and AT-chX-piRNA levels mostly overlap and resemble those for the ping-pong mechanism in the ovaries, Armitage (armi) is not required for the accumulation of AT-chX-1 piRNA. These findings suggest that the impacts of armi mutants on the operation of the piRNA pathway are variable in germ cells of fly testes.


A balance between circular and linear forms of the dengue virus genome is crucial for viral replication. The plasticity of viral plus strand RNA genomes is fundamental for the multiple functions of these molecules. Local and long-range RNA-RNA interactions provide the scaffold for interacting proteins of the translation, replication, and encapsidation machinery. Using dengue virus as a model, we investigated the relevance of the interplay between two alternative conformations of the viral genome during replication. Flaviviruses require long-range RNA-RNA interactions and genome cyclization for RNA synthesis. Here, we define a sequence present in the viral 3'UTR that overlaps two mutually exclusive structures. This sequence can form an extended duplex by long-range 5'-3' interactions in the circular conformation of the RNA or fold locally into a small hairpin (sHP) in the linear form of the genome. A mutational analysis of the sHP structure revealed an absolute requirement of this element for viral viability, suggesting the need of a linear conformation of the genome. Viral RNA replication showed high vulnerability to changes that alter the balance between circular and linear forms of the RNA. Mutations that shift the equilibrium toward the circular or the linear conformation of the genome spontaneously revert to sequences with different mutations that tend to restore the relative stability of the two competing structures. We propose a model in which the viral genome exists in at least two alternative conformations and the balance between these two states is critical for infectivity.


Modified least-variant set normalization for miRNA microarray. MicroRNAs (miRNAs) are short noncoding RNAs that are involved in post-transcriptional regulation of mRNAs. Microarrays have been employed to measure global miRNA expressions; however, because the number of miRNAs is much smaller than the number of mRNAs, it is not clear whether traditional normalization methods developed for mRNA arrays are suitable for miRNA. This is an important question, since normalization affects downstream analyses of the data. In this paper we develop a least-variant set (LVS) normalization method, which was previously shown to outperform other methods in mRNA analysis when standard assumptions are violated. The selection of the LVS miRNAs is based on a robust linear model fit of the probe-level data that takes into account the considerable differences in variances between probes. In a spike-in study, we show that the LVS has similar operating characteristics, in terms of sensitivity and specificity, compared with the ideal normalization, and it is better than no normalization, 75th percentile-shift, quantile, global median, VSN, and lowess normalization methods. We evaluate four expression-summary measures using a tissue data set; summarization from the robust model performs as well as the others. Finally, comparisons using expression data from two dissimilar tissues and two similar ones show that LVS normalization has better operating characteristics than other normalizations.


How I treat superficial venous thrombosis. Thrombosis of superficial veins has long been regarded as a benign disorder. If patients with a clinical diagnosis of superficial venous thrombosis (SVT) are thoroughly evaluated, the degree and extent of thrombosis in patients with SVT are characteristically underestimated (∼ 75% of the time) and such patients have coexistence (∼ 25% of the time) of, and/or rapid progression (∼ 10% of the time) to, systemic venous thromboembolism (VTE). Pulmonary embolism (PE; ∼ 25% of the time) and death (∼ 1% of the time) occur. Contributory risk factors for SVT are the same for VTE. Treatment of patients' SVT with parenteral anticoagulants appears to be both efficacious and certainly safe. I regard most patients with a clinical diagnosis of SVT the same as those with VTEs. Systemic anticoagulant therapy of patients with a clinical diagnosis of SVT obviates extensive imaging and laboratory workup and may be cost effective while encompassing treatment of any unknown concomitant thromboses with only low risk for hemorrhage. This decision is especially clear in those patients with known hypercoagulability. Patients without clinical risk factors are at lower risk to develop VTE complications and might be those who can be simply observed.


Myeloperoxidase attracts neutrophils by physical forces. Recruitment of polymorphonuclear neutrophils (PMNs) remains a paramount prerequisite in innate immune defense and a critical cofounder in inflammatory vascular disease. Neutrophil recruitment comprises a cascade of concerted events allowing for capture, adhesion and extravasation of the leukocyte. Whereas PMN rolling, binding, and diapedesis are well characterized, receptor-mediated processes, mechanisms attenuating the electrostatic repulsion between the negatively charged glycocalyx of leukocyte and endothelium remain poorly understood. We provide evidence for myeloperoxidase (MPO), an abundant PMN-derived heme protein, facilitating PMN recruitment by its positive surface charge. In vitro, MPO evoked highly directed PMN motility, which was solely dependent on electrostatic interactions with the leukocyte's surface. In vivo, PMN recruitment was shown to be MPO-dependent in a model of hepatic ischemia and reperfusion, upon intraportal delivery of MPO and in the cremaster muscle exposed to local inflammation or to intraarterial MPO application. Given MPO's affinity to both the endothelial and the leukocyte's surface, MPO evolves as a mediator of PMN recruitment because of its positive surface charge. This electrostatic MPO effect not only displays a so far unrecognized, catalysis-independent function of the enzyme, but also highlights a principal mechanism of PMN attraction driven by physical forces.


RNA-seq analysis of 2 closely related leukemia clones that differ in their self-renewal capacity. The molecular mechanisms regulating self-renewal of leukemia stem cells remain poorly understood. Here we report the generation of 2 closely related leukemias created through the retroviral overexpression of Meis1 and Hoxa9. Despite their apparent common origin, these clonal leukemias exhibit enormous differences in stem cell frequency (from 1 in 1.4, FLA2; to 1 in 347, FLB1), suggesting that one of these leukemias undergoes nearly unlimited self-renewal divisions. Using next-generation RNA-sequencing, we characterized the transcriptomes of these phenotypically similar, but biologically distinct, leukemias, identifying hundreds of differentially expressed genes and a large number of structural differences (eg, alternative splicing and promoter usage). Focusing on ligand-receptor pairs, we observed high expression levels of Sdf1-Cxcr4; Jagged2-Notch2/1; Osm-Gp130; Scf-cKit; and Bmp15-Tgfb1/2. Interestingly, the integrin beta 2-like gene (Itgb2l) is both highly expressed and differentially expressed between our 2 leukemias (∼ 14-fold higher in FLA2 than FLB1). In addition, gene ontology analysis indicated G-protein-coupled receptor had a much higher proportion of differential expression (22%) compared with other classes (∼ 5%), suggesting a potential role regulating subtle changes in cellular behavior. These results provide the first comprehensive transcriptome analysis of a leukemia stem cell and document an unexpected level of transcriptome variation between phenotypically similar leukemic cells.


Regulation of steady-state neutrophil homeostasis by macrophages. The timely clearance of apoptotic neutrophils from inflammation sites is an important function of macrophages; however, the role of macrophages in maintaining neutrophil homeostasis under steady-state conditions is less well understood. By conditionally deleting the antiapoptotic gene cellular FLICE-like inhibitory protein (C-FLIP) in myeloid cells, we have generated a novel mouse model deficient in marginal zone and bone marrow stromal macrophages. These mice develop severe neutrophilia, splenomegaly, extramedullary hematopoiesis, decreased body weight, and increased production of granulocyte colony-stimulating factor (G-CSF) and IL-1β, but not IL-17. c-FLIP(f/f) LysM-Cre mice exhibit delayed clearance of circulating neutrophils, suggesting that failure of macrophages to efficiently clear apoptotic neutrophils causes production of cytokines that drive excess granulopoiesis. Further, blocking G-CSF but not IL-1R signaling in vivo rescues this neutrophilia, suggesting that a G-CSF-dependent, IL-1β-independent pathway plays a role in promoting neutrophil production in mice with defective clearance of apoptotic cells.


Angiostatic and chemotactic activities of the CXC chemokine CXCL4L1 (platelet factor-4 variant) are mediated by CXCR3. We investigated possible cellular receptors for the human CXC chemokine platelet factor-4 variant/CXCL4L1, a potent inhibitor of angiogenesis. We found that CXCL4L1 has lower affinity for heparin and chondroitin sulfate-E than platelet factor-4 (CXCL4) and showed that CXCL10 and CXCL4L1 could displace each other on microvascular endothelial cells. Labeled CXCL4L1 also bound to CXCR3A- and CXCR3B-transfectants and was displaced by CXCL4L1, CXCL4, and CXCL10. The CXCL4L1 anti-angiogenic activity was blocked by anti-CXCR3 antibodies (Abs) in the Matrigel and cornea micropocket assays. CXCL4L1 application in CXCR3(-/-) or in wild-type mice treated with neutralizing anti-CXCR3 Abs, resulted in reduced inhibitory activity of CXCL4L1 on tumor growth and vascularization of Lewis lung carcinoma. Furthermore, CXCL4L1 and CXCL4 chemoattracted activated T cells, human natural killer cells, and human immature dendritic cells (DCs). Migration of DCs toward CXCL4 and CXCL4L1 was desensitized by preincubation with CXCL10 and CXCL11, inhibited by pertussis toxin, and neutralized by anti-CXCR3 Abs. Chemotaxis of T cells, natural killer cells, and DCs is likely to contribute to the antitumoral action. However, the in vivo data indicate that the angiostatic property of CXCL4L1 is equally important in retarding tumor growth. Thus, both CXCR3A and CXCR3B are implicated in the chemotactic and vascular effects of CXCL4L1.


Metformin decreases GnRH- and activin-induced gonadotropin secretion in rat pituitary cells: potential involvement of adenosine 5' monophosphate-activated protein kinase (PRKA). Metformin is an insulin sensitizer molecule used for the treatment of infertility in women with polycystic ovary syndrome and insulin resistance. It modulates the reproductive axis, affecting the release of gonadotropin-releasing hormone (GnRH) and luteinizing hormone (LH). However, metformin's mechanism of action in pituitary gonadotropin-secreting cells remains unclear. Adenosine 5' monophosphate-activated protein kinase (PRKA) is involved in metformin action in various cell types. Here, we investigated the effects of metformin on gonadotropin secretion in response to activin and GnRH in primary rat pituitary cells (PRP), and studied PRKA in rat pituitary. In PRP, metformin (10 mM) reduced LH and follicle-stimulating hormone (FSH) secretion induced by GnRH (10(-8) M, 3 h), FSH secretion, and mRNA FSHbeta subunit expression induced by activin (10(-8) M, 12 or 24 h). The different subunits of PRKA are expressed in pituitary. In particular, PRKAA1 is detected mainly in gonadotrophs and thyrotrophs, is less abundant in lactotrophs and somatotrophs, and is undetectable in corticotrophs. In PRP, metformin increased phosphorylation of both PRKA and acetyl-CoA carboxylase. Metformin decreased activin-induced SMAD2 phosphorylation and GnRH-induced mitogen-activated protein kinase (MAPK) 3/1 (ERK1/2) phosphorylation. The PRKA inhibitor compound C abolished the effects of metformin on gonadotropin release induced by GnRH and on FSH secretion and Fshb mRNA induced by activin. The adenovirus-mediated production of dominant negative PRKA abolished the effects of metformin on the FSHbeta subunit mRNA and SMAD2 phosphorylation induced by activin and on the MAPK3/1 phosphorylation induced by GnRH. Thus, in rat pituitary cells, metformin decreases gonadotropin secretion and MAPK3/1 phosphorylation induced by GnRH and FSH release, FSHbeta subunit expression, and SMAD2 phosphorylation induced by activin through PRKA activation.


Vascular bed-specific regulation of the von Willebrand factor promoter in the heart and skeletal muscle. A region of the human von Willebrand factor (VWF) gene between -2812 and the end of the first intron (termed vWF2) was previously shown to direct expression in the endothelium of capillaries and a subset of larger blood vessels in the heart and skeletal muscle. Here, our goal was to delineate the DNA sequences responsible for this effect. A series of constructs containing deletions or mutations of vWF2 coupled to LacZ were targeted to the Hprt locus of mice, and the resulting animals were analyzed for reporter gene expression. The findings demonstrate that DNA sequences between -843 and -620 are necessary for expression in capillary but not large vessel endothelium in heart and skeletal muscle. Further, expression of VWF in capillaries and larger vessels of both tissues required the presence of a native or heterologous intron. In vitro assays implicated a role for ERG-binding ETS motif at -56 in mediating basal expression of VWF. In Hprt-targeted mice, mutation of the ETS consensus motif resulted in loss of LacZ expression in the endothelium of the heart and skeletal muscle. Together, these data indicate that distinct DNA modules regulate vascular bed-specific expression of VWF.


The effects of estrogen and progesterone on blood glutamate levels: evidence from changes of blood glutamate levels during the menstrual cycle in women. The gonadal steroids estrogen and progesterone have been shown to have neuroprotective properties against various neurodegenerative conditions. Excessive concentrations of glutamate have been found to exert neurotoxic properties. We hypothesize that estrogen and progesterone provide neuroprotection by the autoregulation of blood and brain glutamate levels. Venous blood samples (10 ml) were taken from 31 men and 45 women to determine blood glutamate, estrogen, progesterone, glucose, glutamate-pyruvate transaminase (GPT), and glutamate-oxaloacetate transaminase (GOT) levels, collected on Days 1, 7, 12, and 21 of the female participants' menstrual cycle. Blood glutamate concentrations were higher in men than in women at the start of menstruation (P < 0.05). Blood glutamate levels in women decreased significantly on Days 7 (P < 0.01), 12 (P < 0.001), and 21 (P < 0.001) in comparison with blood glutamate levels on Day 1. There was a significant decrease in blood glutamate levels on Days 12 (P < 0.001) and 21 (P < 0.001) in comparison with blood glutamate levels on Day 7. Furthermore, there was an increase in blood glutamate levels on Day 21 compared with Day 12 (P < 0.05). In women, there were elevated levels of estrogen on Days 7 (P < 0.05), 12, and 21 (P < 0.001), and elevated levels of progesterone on Days 12 and 21 (P < 0.001). There were no differences between men and women with respect to blood glucose concentrations. Concentrations of GOT (P < 0.05) and GPT (P < 0.001) were significantly higher in men than in women during the entire cycle. The results of this study demonstrate that blood glutamate levels are inversely correlated to levels of plasma estrogen and progesterone.


Follicular fluid high-density lipoprotein-associated sphingosine 1-phosphate (S1P) promotes human granulosa lutein cell migration via S1P receptor type 3 and small G-protein RAC1. Coordinated migration and progesterone production by granulosa cells is critical to the development of the corpus luteum, but the underlying mechanisms remain obscure. Sphingosine 1-phosphate (S1P), which is associated with follicular fluid high-density lipoprotein (FF-HDL), was previously shown to regulate ovarian angiogenesis. We herein examined the effects of S1P and FF-HDL on the function of granulosa lutein cells. Both FF-HDL and S1P induced migration of primary human granulosa lutein cells (hGCs) and the granulosa lutein cell line HGL5. In addition, FF-HDL but not S1P promoted progesterone synthesis, and neither of the two compounds stimulated proliferation of granulosa lutein cells. Polymerase chain reaction and Western blot experiments demonstrated the expression of S1P receptor type 1 (S1PR1), S1PR2, S1PR3, and S1PR5 but not S1PR4 in hGCs and HGL5 cells. The FF-HDL- and S1P-induced granulosa lutein cell migration was emulated by FTY720, an agonist of S1PR1, S1PR3, S1PR4, and S1PR5, and by VPC24191, an agonist of S1PR1 and S1PR3, but not by SEW2871 and phytosphingosine 1-phosphate, agonists of S1PR1 and S1PR4, respectively. In addition, blockade of S1PR3 with CAY1044, suramine, or pertussis toxin inhibited hGC and HGL5 cell migration toward FF-HDL or S1P, while blockade of S1PR1 and S1PR2 with W146 and JTE013, respectively, had no effect. Both FF-HDL and S1P triggered activation of small G-protein RAC1 and actin polymerization in granulosa cells, and RAC1 inhibition with Clostridium difficile toxin B or NSC23766 abolished FF-HDL- and S1P-induced migration. The FF-HDL-associated S1P promotes granulosa lutein cell migration via S1PR3 and RAC1 activation. This may represent a novel mechanism contributing to the development of the corpus luteum.


Survival and size are differentially regulated by placental and fetal PKBalpha/AKT1 in mice. The importance of placental circulation is exemplified by the correlation of placental size and blood flow with fetal weight and survival during normal and compromised human pregnancies in such conditions as preeclampsia and intrauterine growth restriction (IUGR). Using noninvasive magnetic resonance imaging, we evaluated the role of PKBalpha/AKT1, a major mediator of angiogenesis, on placental vascular function. PKBalpha/AKT1 deficiency reduced maternal blood volume fraction without affecting the integrity of the fetomaternal blood barrier. In addition to angiogenesis, PKBalpha/AKT1 regulates additional processes related to survival and growth. In accordance with reports in adult mice, we demonstrated a role for PKBalpha/AKT1 in regulating chondrocyte organization in fetal long bones. Using tetraploid complementation experiments with PKBalpha/AKT1-expressing placentas, we found that although placental PKBalpha/AKT1 restored fetal survival, fetal PKBalpha/AKT1 regulated fetal size, because tetraploid complementation did not prevent intrauterine growth retardation. Histological examination of rescued fetuses showed reduced liver blood vessel and renal glomeruli capillary density in PKBalpha/Akt1 null fetuses, both of which were restored by tetraploid complementation. However, bone development was still impaired in tetraploid-rescued PKBalpha/Akt1 null fetuses. Although PKBalpha/AKT1-expressing placentas restored chondrocyte cell number in the hypertrophic layer of humeri, fetal PKBalpha/AKT1 was found to be necessary for chondrocyte columnar organization. Remarkably, a dose-dependent phenotype was exhibited for PKBalpha/AKT1 when examining PKBalpha/Akt1 heterozygous fetuses as well as those complemented by tetraploid placentas. The differential role of PKBalpha/AKT1 on mouse fetal survival and growth may shed light on its roles in human IUGR.


Polypyrimidine tract-binding protein 2 binds to selective, intronic messenger RNA and microRNA targets in the mouse testis. Here we use an in vivo cross-linking and immunoprecipitation procedure to detect RNA targets of the multifunctional RNA-binding protein polypyrimidine tract-binding protein (PTBP) 2 in mouse testis. Eleven known mRNAs, including Ptbp2 mRNA, 28 RNAs matching intron sequences, and 12 small RNAs and repeat sequences are identified. The specificity of interaction between PTBP2 and its target RNAs was confirmed using RNA interference with mouse N2A cells. Reduction of PTBP2 levels led to decreases in 7 of 10 of the mRNAs, to the repression of alternative splicing of introns, and to reductions in specific miRNAs.


The role of nutrition in the regulation of luteinizing hormone secretion by the opioidergic, dopaminergic, and serotonergic systems in female Mediterranean goats. This study examined which neural mechanism (opioid, dopaminergic, or serotonergic system) is involved in the regulation of luteinizing hormone (LH) secretion, with and without nutritional modulation, at different times of the photoperiodic cycle. Goats were randomly distributed into two experimental groups that received either 1.1 (high group; n = 18) or 0.7 (low group; n = 18) times the nutritional maintenance requirements. The goats were exposed to alternations of 3 mo of long days and 3 mo of short days. Plasma LH concentrations were measured twice a week. The effects of intravenous injections of naloxone (endogenous opioid receptor antagonist), pimozide (dopaminergic(2) receptor antagonist), and cyproheptadine (serotonin 5-hydroxytryptamine(2) receptor antagonist) on LH secretion were assessed during challenges in three different photoperiodic situations: the onset of LH stimulation by short days (OnsetSD), the onset of LH inhibition by long days (OnsetLD), and during the LH inhibition by long days (LateLD). The role of the different neural systems was clearly modified by the level of nutrition. In the low-nutrition group, only naloxone increased LH concentrations during onsetLD (P < 0.05). However, in the high-nutrition group, naloxone increased the concentration and pulsatility of LH (P < 0.05) in onsetSD and onsetLD. Pimozide increased LH concentration and pulsatility (P < 0.05) in onsetLD and LH concentration in lateLD (P < 0.001). Finally, cyproheptadine significantly increased LH concentration at all three times (P < 0.001). These results provide evidence that all three systems are involved in the inhibition of LH release in onsetLD, and that the opioid and serotonin mechanisms are involved during the onsetSD that were enhanced by a high plane of nutrition.


Thrombophilic-type placental pathologies and skeletal growth delay following maternal administration of angiostatin4.5 in mice. During placentation, the concentration of fibrinous deposits on the surfaces of maternal vasculature plays a role in villous development and has been strongly implicated in the pathophysiology of human fetal growth restriction (FGR). Fibrinous deposits are conspicuous sites of platelet aggregation where there is local activation of the hemostatic cascade. During activation of the hemostatic cascade, a number of pro- and antiangiogenic agents may be generated at the cell surface, and an imbalance in these factors may contribute to the placental pathology characteristic of FGR. We tested the hypothesis that angiostatin(4.5) (AS(4.5)), a cleavage fragment of plasminogen liberated at the cell surface, is capable of causing FGR in mice. Increased maternal levels of AS(4.5) in vivo result in reproducible placental pathology, including an altered vascular compartment (both in decidual and labyrinthine layers) and increased apoptosis throughout the placenta. In addition, there is significant skeletal growth delay and conspicuous edema in fetuses from mothers that received AS(4.5). Maternally generated AS(4.5), therefore, can access maternal placental vasculature and have a severe effect on placental architecture and inhibit fetal development in vivo. These findings strongly support the hypothesis that maternal AS(4.5) levels can influence placental development, possibly by directly influencing trophoblast turnover in the placenta, and contribute to fetal growth delay in mice.


Asynchronous expression of the homeodomain protein CUX1 in Sertoli cells and spermatids during spermatogenesis in mice. The homeodomain CUX1 protein exists as multiple isoforms that arise from proteolytic processing of a 200-kDa protein or an alternate splicing or from the use of an alternate promoter. The 200-kDa CUX1 protein is highly expressed in the developing kidney, where it functions to regulate cell proliferation. Transgenic mice ectopically expressing the 200-kDa CUX1 protein develop renal hyperplasia associated with reduced expression of the cyclin kinase inhibitor p27. A 55-kDa CUX1 isoform is expressed exclusively in the testes. We determined the pattern and timing of CUX1 protein expression in developing testes. CUX1 expression was continuous in Sertoli cells from prepubertal testes but became cyclic when spermatids appeared. In testes from mature mice, CUX1 was highly expressed only in round spermatids at stages IV-V of spermatogenesis, in both spermatids and Sertoli cells at stages VI-X of spermatogenesis, and only in Sertoli cells at stage XI of spermatogenesis. While most of the seminiferous tubules in wild-type mice were between stages VI and X of spermatogenesis, there was a significant reduction in the percentage of seminiferous tubules between stages VI and X in Cux1 transgenic mice and a significant increase in the percentage of seminiferous tubules in stages IV-V and XI. Moreover, CUX1 was not expressed in proliferating cells in testes from either wild-type or transgenic mice. Thus, unlike the somatic form of CUX1, which has a role in cell proliferation, the testis-specific form of CUX1 is not involved in cell division and appears to play a role in signaling between Sertoli cells and spermatids.


How desirable are your IC50s? A way to enhance screening-based decision making. Dose-response curves, resulting in estimates of endpoints such as the IC(50), are fundamental to drug discovery. However, some estimates are more reliable than others. It is important to know just how reliable an estimate is if we want to base decisions on it or use it in further modeling. In this study, the authors propose a new measure of endpoint reliability, based on the concept of desirability first introduced by Harrington. The solution is not dependent on the application used to analyze the experimental data, provided a number of parameters to characterize the dose-response curve are available. The authors show how this score can be used as an objective and consistent measure to rank screening results, combine information from groups of experiments, and determine optimal levels of characterization of a compound's biological activity.


IGF2 actions on trophoblast in human placenta are regulated by the insulin-like growth factor 2 receptor, which can function as both a signaling and clearance receptor. Insulin-like growth factor 2 (IGF2) enhances proliferation and survival of human first-trimester cytotrophoblasts (CTB) by signaling through the insulin-like growth factor 1 receptor (IGF1R). However, the role of the IGF2 receptor (IGF2R) in regulating trophoblast kinetics is unclear: It could act as a clearance receptor for trafficking excess ligand to lysosomes for degradation and/or directly mediate IGF2 signaling. We used an IGF2R knockdown strategy in BeWo cells and placental villous explants to investigate trophoblast proliferation and survival in response to stimulation by IGF. Both IGF1 and IGF2 significantly (P < 0.001) increased mitosis and reduced apoptosis in serum-starved BeWo cells. Small interfering RNA (siRNA)-mediated knockdown of IGF2R further enhanced IGF2-stimulated mitosis (P < 0.01), and IGF2-mediated rescue of apoptosis (P < 0.001) in these cells. Leu(27)IGF2, an IGF2 analogue that binds to IGF2R but not IGF1R, also protected IGF2R-expressing BeWo cells from apoptosis but did not increase mitosis. IGF treatment of term placental villous explants with reduced syncytial expression of IGF2R increased CTB proliferation (P < 0.001) and decreased apoptosis (P < 0.01) compared to untreated controls. Moreover, IGF2-mediated rescue of CTB apoptosis was significantly greater than that in tissue with normal IGF2R expression. Leu(27)IGF2 promoted mitogenesis and survival only in explants with intact IGF2R expression. Given that altered CTB turnover is observed in pregnancies complicated by fetal growth restriction, the development of strategies to manipulate the IGF2R signaling axis in the syncytiotrophoblast may provide a therapeutic avenue for treating this condition.


ICOStomizing immunotherapies with T(H)17. New insights on the role of costimulatory molecules in T helper cell function have yielded exciting alternatives to the development of therapeutic strategies that target T cell costimulatory pathways. Inducible costimulatory molecule (ICOS) signaling is now shown by Paulos and colleagues to support expansion of human T helper 17 (T(H)17) cells that could exert antitumor activity. Here we discuss (i) how these findings aid in our understanding of mechanisms that govern T(H)17 cell functions and (ii) the potential application of these new insights to the development of immunotherapies.


The inducible costimulator (ICOS) is critical for the development of human T(H)17 cells. Human T helper 17 (T(H)17) cells regulate host defense, autoimmunity, and tumor immunity. Although cytokines that control human T(H)17 cell development have been identified, the costimulatory molecules important for T(H)17 cell generation are unknown. Here, we found that the inducible costimulator (ICOS) was critical for the differentiation and expansion of human T(H)17 cells. Human cord blood contained a subset of CD161(+)CD4(+) T cells that were recent emigrants from the thymus, expressed ICOS constitutively, and were imprinted as T(H)17 cells through ICOS signaling. ICOS stimulation induced c-MAF, RORC2, and T-bet expression in these cells, leading to increased secretion of interleukin-21 (IL-21), IL-17, and interferon-γ (IFN-γ) compared with cells stimulated with CD28. Conversely, CD28 ligation abrogated ICOS costimulation, dampening RORC2 expression while promoting the expression of the aryl hydrocarbon receptor, which led to reduced secretion of IL-17 and enhanced production of IL-22 compared with cells stimulated with ICOS. Moreover, ICOS promoted the robust expansion of IL-17(+)IFN-γ(+) human T cells, and the antitumor activity of these cells after adoptive transfer into mice bearing large human tumors was superior to that of cells expanded with CD28. The therapeutic effectiveness of ICOS-expanded cells was associated with enhanced functionality and engraftment in vivo. These findings reveal a vital role for ICOS signaling in the generation and maintenance of human T(H)17 cells and suggest that components of this pathway could be therapeutically targeted to treat cancer or chronic infection and, conversely, that interruption of this pathway may have utility in multiple sclerosis and other autoimmune syndromes. These findings have provided the rationale for designing new clinical trials for tumor immunotherapy.


Thermal enhancement with optically activated gold nanoshells sensitizes breast cancer stem cells to radiation therapy. Breast cancer metastasis and disease recurrence are hypothesized to result from residual cancer stem cells, also referred to as tumor-initiating cells, which evade initial treatment. Using both syngeneic mouse and human xenograft models of triple-negative breast cancer, we have demonstrated that a subpopulation enriched in cancer stem cells was more resistant to treatment with 6 gray of ionizing radiation than the bulk of the tumor cells, and accordingly their relative proportion increased 48 to 72 hours after ionizing radiation treatment. In contrast, we achieved a larger reduction in tumor size without a concomitant increase in the percentage of cancer stem cells by treating with local hyperthermia for 20 minutes at 42°C after ionizing radiation using intravenously administered, optically activated gold nanoshells. Forty-eight hours after treatment, cells derived from the tumors treated with ionizing radiation plus hyperthermia exhibited both a marked decrease in tumorigenicity and a more differentiated phenotype than mock- and ionizing radiation-treated tumors. Thus, we have confirmed that these cancer stem cells are responsible for accelerated repopulation in vivo and demonstrated that hyperthermia sensitizes this cell population to radiation treatment. These findings suggest that local hyperthermia delivered by gold nanoshells plus radiation can eliminate radioresistant breast cancer stem cells.


Moving to place: childhood cancer treatment decision making in single-parent and repartnered family structures. Few researchers have studied how parents from diverse family structures cope with childhood chronic illness. We designed this study to discern the childhood cancer treatment decision-making (TDM) process in these families. Using grounded theory, we interviewed 15 custodial parents, nonresidential parents, and stepparents who had previously made a major treatment decision for their children with cancer. "Moving to place" was the central psychosocial process by which parents negotiated involvement in TDM. Parents moved toward or were moved away from involvement based on parent position in the family (custodial, nonresidential, and stepparent), prediagnosis family dynamics, and time since diagnosis. Parents used the actions of stepping up, stepping back, being pushed, and stepping away to respond to the need for TDM. Parents faced additional stressors because of their family situations, which affected the TDM process. Findings from this study provide important insight into diverse families and their unique parental TDM experiences.


Preserving the self: the process of decision making about hereditary breast cancer and ovarian cancer risk reduction. Women who carry BRCA1 or BRCA2 (BRCA1/2) gene mutations have up to an 88% lifetime risk of breast cancer and up to a 65% lifetime risk of ovarian cancer. Strategies to address these risks include cancer screening and risk-reducing surgery (i.e., mastectomy and salpingo-oophorectomy). We conducted a grounded theory study with 22 BRCA1/2 mutation-carrier women to understand how women make decisions about these risk-reducing strategies. Preserving the self was the overarching decision-making process evident in the participants' descriptions. This process was shaped by contextual conditions including the characteristics of health services, the nature of hereditary breast and ovarian cancer risk-reduction decisions, gendered roles, and the women's perceived proximity to cancer. The women engaged in five decision-making styles, and these were characterized by the use of specific decision-making approaches. These findings provide theoretical insights that could inform the provision of decisional support to BRCA1/2 carriers.


Examination of the factor structure and concurrent validity of the Langer Mindfulness/Mindlessness Scale. Langer's theory of mindfulness proposes that a mindful person seeks out and produces novelty, is attentive to context, and is flexible in thought and behavior. In three independent studies, the factor structure of the Langer Mindfulness/Mindlessness Scale was examined. Confirmatory factor analysis failed to replicate the four-factor model and a subsequent exploratory factor analysis revealed the presence of a two-factor (mindfulness and mindlessness) solution. Study 2 demonstrated that the two factors assessed discrete constructs and were not merely products of acquiescence. Support was also found for a nine-item, one-factor model comprised solely of mindfulness items. On comparing models, Study 3 suggested the superiority of the one-factor mindfulness model. Finally, a preliminary investigation of the concurrent validity of the revised nine-item Langer Mindfulness/Mindlessness Scale is presented. The current article offers researchers a revised version of a mindfulness measure derived from a cognitive perspective.


The Bermond-Vorst Alexithymia Questionnaire: a measurement invariance examination among U.S. Anglos and U.S. Hispanics. The aim of this article is to assess the measurement invariance of the Bermond-Vorst Alexithymia Questionnaire (BVAQ) in U.S. Anglo (n = 490) and U.S. Hispanic ( n = 379) samples of college students. The BVAQ items demonstrated invariance of the factor loadings, the latent item intercepts, and unique factor variances. However, Hispanics had higher verbalizing scores and lower fantasizing scores on the latent variables. As a whole, the BVAQ requires additional testing and/or refinement for use with a Hispanic sample. Until such refinements, the authors recommend that the Toronto Alexithymia Scale-20 be used with U.S. Hispanic samples.


Corneal dystrophies: molecular genetics to therapeutic intervention--Fifth ARVO/Pfizer Ophthalmics Research Institute Conference.


Inflammation in an individual joint predicts damage to that joint in psoriatic arthritis. The authors have previously reported on the relationship between activity and subsequent damage at the patient level for patients with psoriatic arthritis (PsA). The aim of this study was to identify key predictors of damage to individual joints in the hands and feet of patients with PsA, in particular those that capture previous activity. Data from patients followed prospectively at the University of Toronto PsA clinic between 1978 and 2006 were available for analysis. Logistic regression was used to relate the probability of a joint developing damage, within a specified time interval after the most recent clinic visit, to potential predictor variables. The predictor variables considered encompassed the history of disease activity of the joint and elsewhere, previous damage and the timing of clinical assessments. 511 patients with no hand damage at clinic entry and 552 patients with no foot damage at clinic entry were included in the analysis of the hand and foot joints, respectively. The analysis of the hand and foot joints demonstrated that the activity (tenderness and/or swelling) history of the specific joint is associated with subsequent damage. For the joints of the feet, activity observations elsewhere in the same foot, and in particular the same toe, were also shown to be associated with subsequent damage. Both joint tenderness and swelling are important predictors of joint damage in PsA.


Vitamin D: an instrumental factor in the anti-phospholipid syndrome by inhibition of tissue factor expression. Antiphospholipid syndrome (APS) is a systemic autoimmune disease characterised by thrombosis, obstetric complications and the presence of anti-phospholipid antibodies such as anti-β2GPI-Abs. These antibodies may set off the coagulation cascade via several mechanisms, including the induction of tissue factor (TF) expression. Vitamin D has recently emerged as an immunomodulator that might exert an anti-thrombotic effect. Therefore, we studied serum vitamin D levels in a cohort of APS patients, as well as the effect of vitamin D in an in vitro model of APS-mediated thrombosis. Serum vitamin D levels were measured in 179 European APS patients and 141 healthy controls using the LIAISON chemiluminescent immunoassay, and the levels were evaluated in conjunction with a wide spectrum of clinical manifestations. In an vitro model, anti-β2GPI antibodies were purified from four patients with APS to evaluate the expression of TF in activated starved human umbilical vein endothelial cells. The effect of vitamin D (1,25-dihydroxyvitamin D, 10 nm) on anti-β2GPI-Abs mediated TF expression was analysed by immunoblot. Vitamin D deficiency (serum level ≤15 ng/ml) was documented in 49.5% of our APS patients versus 30% of controls (p<0.001) and was significantly correlated with thrombosis (58% vs 42%; p<0.05), neurological and ophthalmic manifestations, pulmonary hypertension, livedo reticularis and skin ulcerations. In vitro vitamin D inhibited the expression of TF induced by anti-β2GPI-antibodies. Vitamin D deficiency is common among APS patients and is associated with clinically defined thrombotic events. Vitamin D inhibits anti-β2GPI-mediated TF expression in vitro. Thus, vitamin D deficiency might be associated with decreased inhibition of TF expression and increased coagulation in APS. Evaluation of vitamin D status and vitamin D supplementation in APS patients should be considered.


Ophthalmic drug delivery systems for the treatment of retinal diseases: basic research to clinical applications.


Sociocognitive self-regulatory mechanisms governing judgments of the acceptability and likelihood of sport cheating. This study extends previous psychosocial literature (Bandura et al., 2001, 2003) by examining a structural model of the self-regulatory mechanisms governing the acceptability and likelihood of cheating in a sport context. Male and female adolescents (N = 804), aged 15-20 years, took part in this study. Negative affective self-regulatory efficacy influenced the acceptability and likelihood of cheating through the mediating role of moral disengagement, in females and males. Affective efficacy positively influenced prosocial behavior through moral disengagement or through resistive self-regulatory efficacy and social efficacy, in both groups. The direct effects of affective efficacy on beliefs about cheating were only evident in females. These results extend the findings of Bandura et al. (2001, 2003) to the sport context and suggest that affective and resistive self-regulatory efficacy operate in concert in governing adolescents' moral disengagement and transgressive behaviors in sport.


Fcγ receptor IIIb polymorphism and use of glucocorticoids at baseline are associated with infusion reactions to infliximab in patients with rheumatoid arthritis. Infusion reaction is a major adverse event in patients with rheumatoid arthritis (RA) treated with infliximab. The possible factors including Fcγ receptor (FcγR) polymorphism associated with the development of infusion reactions in patients with RA receiving infliximab were prospectively examined. 96 patients with RA were enrolled and scheduled to receive infliximab at a dose of 3 mg/kg at weeks 0, 2 and 6 and every 8 weeks thereafter. Genetic polymorphisms for FcγR were examined in FCGR3A 176F/V and FCGR3B NA1/2 alleles by allele-specific PCR analysis. An infusion reaction was observed in 17 patients (18%) during 52 weeks of treatment with infliximab. The FCGR3B NA1/NA1 genotype was found in 75% of the patients with infusion reactions and in only 37% of those without (p=0.01), whereas the FCGR3A 176F/V genotype was equally distributed in the patients with or without infusion reactions. Glucocorticoids were used in 53% of the patients who developed an infusion reaction and in 80% of those without an infusion reaction (p=0.02). A multivariable logistic regression model showed that the FCGR3B NA1/NA1 genotype and use of glucocorticoids at baseline could be used as independent predictive factors for infusion reactions (OR 6.1 (95% CI 1.9 to 24.3) and OR 0.26 (95% CI 0.08 to 0.84), respectively). The presence of anti-infliximab antibody during infliximab treatment was also associated with infusion reactions. FCGR3B NA1/NA1 genotype, use of glucocorticoids and the presence of anti-infliximab antibody accounted for nearly all patients with RA who developed infusion reactions.


How you provide corrective feedback makes a difference: the motivating role of communicating in an autonomy-supporting way. We relied on self-determination theory (SDT; Deci & Ryan, 2000) to investigate to what extent autonomy-supporting corrective feedback (i.e., feedback that coaches communicate to their athletes after poor performance or mistakes) is associated with athletes' optimal motivation and well-being. To test this hypothesis, we conducted a cross-sectional study with 337 (67.1% males) Greek adolescent athletes (age M = 15.59, SD = 2.37) from various sports. Aligned with SDT, we found through path analysis that an autonomy-supporting versus controlling communication style was positively related to future intentions to persist and well-being and negatively related to ill-being. These relations were partially mediated by the perceived legitimacy of the corrective feedback (i.e., the degree of acceptance of corrective feedback), and, in turn, by intrinsic motivation, identified regulation, and external regulation for doing sports. Results indicate that autonomy-supporting feedback can be still motivating even in cases in which such feedback conveys messages of still too low competence.


Developmental activities that contribute to high or low performance by elite cricket batters when recognizing type of delivery from bowlers' advanced postural cues. We examined the developmental activities that contribute to the development of superior anticipation skill among elite cricket batters. The batters viewed 36 video clips involving deliveries from bowlers that were occluded at ball release and were required to predict delivery type. Accuracy scores were used to create two subgroups: high-performing and low-performing anticipators. Questionnaires were used to record the participation history profiles of the groups. In the early stages of development, hours accumulated in cricket and other sports, as well as milestones achieved, did not differentiate groups. Significant between-group differences in activity profiles were found between 13 and 15 years of age, with high-performing anticipators accumulating more hours in structured cricket activity, and specifically in batting, compared with their low-performing counterparts.


Cultural differences in the relationships among autonomy support, psychological need satisfaction, subjective vitality, and effort in British and Chinese physical education. Using basic psychological needs theory (BPNT; Ryan & Deci, 2000) as our guiding framework, we explored cultural differences in the relationships among physical education students' perceptions of teacher autonomy support, psychological need satisfaction, subjective vitality and effort in class. Seven hundred and fifteen students (age range from 13 to 15 years) from the U.K. and Hong Kong, China, completed a multisection inventory during a timetabled physical education class. Multilevel analyses revealed that the relationships among autonomy support, subjective vitality and effort were mediated by students' perceptions of psychological need satisfaction. The relationship between autonomy support and perceptions of competence was stronger in the Chinese sample, compared with the U.K. sample. In addition, the relationship between perceptions of relatedness and effort was not significant in the Chinese students. The findings generally support the pan-cultural utility of BPNT and imply that a teacher-created autonomy supportive environment may promote positive student experiences in both cultures.


Momentary affect predicts bodily movement in daily life: an ambulatory monitoring study. There is converging evidence that physical activity influences affective states. It has been found that aerobic exercise programs can significantly diminish negative affect. Moreover, among healthy individuals, moderate levels of physical activity seem to increase energetic arousal and positive affect. However, the predictive utility of affective states for bodily movement has rarely been investigated. In this study, we examined whether momentarily assessed affect is associated with bodily movement in everyday life. Using a previously published data set (Schwerdtfeger, Eberhardt, & Chmitorz, 2008), we reanalyzed 12-hr ecological momentary assessment (EMA) data from 124 healthy volunteers. Electronic momentary positive-activated affect (EMA-PAA) and negative affect (EMA-NA) were assessed via handheld computers, and bodily movement was recorded via accelerosensors. Generalized linear mixed models were calculated. Results indicated that EMAPAA increases were accompanied by bodily movement increases of varying intensity. EMA-NA was also positively associated with increases in certain kinds of bodily movement. In light of previous research, this finding suggests that affect and bodily movement may have circular effects on each other.


Predictors of doping intentions in elite-level athletes: a social cognition approach. Doping use is an ongoing problem in contemporary sports. Despite efforts to detect and control doping, research on its etiology is limited, especially among elite-level athletes. The present study used an integrated social cognition model to examine the predictors of doping intentions. Structured anonymous questionnaires were completed by 1075 Greek adult elite-level athletes (M age = 25 years, SD = 5.89, 36.1% females) from both team and individual sports. Multiple regression and mediation analyses showed that attitudes, normative beliefs, situational temptation, and behavioral control significantly predicted doping intentions. A normative process was identified whereby situational temptation mediated the effects of normative beliefs on intentions. The findings provide the basis for future social cognition research in doping use, and set the framework for the development of evidence-based preventive interventions.


Beyond sensation seeking: affect regulation as a framework for predicting risk-taking behaviors in high-risk sport. Sensation seeking has been widely studied when investigating individual differences in the propensity for taking risks. However, risk taking can serve many different goals beyond the simple management of physiological arousal. The present study is an investigation of affect self-regulation as a predictor of risk-taking behaviors in high-risk sport. Risk-taking behaviors, negative affectivity, escape self-awareness strategy, and sensation seeking data were obtained from 265 high-risk sportsmen. Moderated hierarchical regression analysis revealed significant main and interaction effects of negative affectivity and escape self-awareness strategy in predicting risk-taking behaviors: high-risk sportsmen's negative affectivity leads them to adopt risk-taking behaviors only if they also use escape self-awareness strategy. Furthermore, the affective model remained significant when controlling for sensation seeking. The present study contributes to an in-depth understanding of risk taking in high-risk sport.


The exercise and affect relationship: evidence for the dual-mode model and a modified opponent process theory. This study examined the relationship between exertion level and affect using the framework of opponent-process theory and the dual-mode model, with the Activation-Deactivation Adjective Checklist and the State Anxiety Inventory among 14 active and 14 sedentary participants doing 20 min of treadmill exercise at speeds of 5% below, 5% above, and at lactate threshold (LT). We found a significant effect of time, condition, Time × Condition, and Time × Group, but no group, Group × Condition, or Time × Group × Condition effects, such that the 5% above LT condition produced a worsening of affect in-task compared with all other conditions whereas, across conditions, participants experienced in-task increases in energy and tension, and in-task decreases in tiredness and calmness relative to baseline. Posttask, participants experienced mood improvement (decreased tension, anxiety, and increased calmness) across conditions, with a 30-min delay in the above LT condition. These results partially support the dual-mode model and a modified opponent-process theory.


PathJam: a new service for integrating biological pathway information. Biological pathways are crucial to much of the scientific research today including the study of specific biological processes related with human diseases. PathJam is a new comprehensive and freely accessible web-server application integrating scattered human pathway annotation from several public sources. The tool has been designed for both (i) being intuitive for wet-lab users providing statistical enrichment analysis of pathway annotations and (ii) giving support to the development of new integrative pathway applications. PathJam’s unique features and advantages include interactive graphs linking pathways and genes of interest, downloadable results in fully compatible formats, GSEA compatible output files and a standardized RESTful API.


Improved elimination of motion artifacts from a photoplethysmographic signal using a Kalman smoother with simultaneous accelerometry. A photoplethysmography (PPG) signal provides very useful information about a subject's hemodynamic status in a hospital or ubiquitous environment. However, PPG is very vulnerable to motion artifacts, which can significantly distort the information belonging to the PPG signal itself. Thus, the reduction of the effects of motion artifacts is an important issue when monitoring the cardiovascular system by PPG. There have been many adaptive techniques to reduce motion artifacts from PPG signals. In the present study, we compared a method based on the fixed-interval Kalman smoother with the usual adaptive filtering algorithms, e.g. the normalized least mean squares, recursive least squares and the conventional Kalman filter. We found that the fixed-interval Kalman smoother reduced motion artifacts from the PPG signal most effectively. Therefore, the use of the fixed-interval Kalman smoother can reduce motion artifacts in PPG, thus providing the most reliable information that can be deduced from the reconstructed PPG signals.


The removal of EMG in EEG by neural networks. In this paper, it is presented that electromyography (EMG) is a shot noise based on the generation of EMG. A novel filter is proposed by applying a neural network (NN) ensemble where the noisy input signal and the desired one are the same in a learning process. Both incremental and batch mode are applied in the learning process of NNs that is better than generalized NN filters. This NN ensemble filter not only reduces additive and multiplicative white noise inside signals, but also preserves the signals' characteristics. In clinical EEG and EMG signals processing, the filter is capable of reducing EMG in the clinical EEG, and it is proved that there is randomness in EMG.


Factors influencing the mechanical behaviour of healthy human descending thoracic aorta. In recent times, significant effort has been made to understand the mechanical behaviour of the arterial wall and how it is affected by the different vascular pathologies. However, to be able to interpret the results correctly, it is essential that the influence of other factors, such as aging or anisotropy, be understood. Knowledge of mechanical behaviour of the aorta has been customarily constrained by lack of data on fresh aortic tissue, especially from healthy young individuals. In addition, information regarding the point of rupture is also very limited. In this study, the mechanical behaviour of the descending thoracic aorta of 28 organ donors with no apparent disease, whose ages vary from 17 to 60 years, is evaluated. Tensile tests up to rupture are carried out to evaluate the influence of age and wall anisotropy. Results reveal that the tensile strength and stretch at failure of healthy descending aortas show a significant reduction with age, falling abruptly beyond the age of 30. This fact places age as a key factor when mechanical properties of descending aorta are considered.


Post-traumatic stress disorder: a right temporal lobe syndrome? In a recent paper (Georgopoulos et al 2010 J. Neural Eng. 7 016011) we reported on the power of the magnetoencephalography (MEG)-based synchronous neural interactions (SNI) test to differentiate post-traumatic stress disorder (PTSD) subjects from healthy control subjects and to classify them with a high degree of accuracy. Here we show that the main differences in cortical communication circuitry between these two groups lie in the miscommunication of temporal and parietal and/or parieto-occipital right hemispheric areas with other brain areas. This lateralized temporal-posterior pattern of miscommunication was very similar but was attenuated in patients with PTSD in remission. These findings are consistent with observations (Penfield 1958 Proc. Natl Acad. Sci. USA 44 51-66, Penfield and Perot 1963 Brain 86 595-696, Gloor 1990 Brain 113 1673-94, Banceaud et al 1994 Brain 117 71-90, Fried 1997 J. Neuropsychiatry Clin. Neurosci. 9 420-8) that electrical stimulation of the temporal cortex in awake human subjects, mostly in the right hemisphere, can elicit the re-enactment and re-living of past experiences. Based on these facts, we attribute our findings to the re-experiencing component of PTSD and hypothesize that it reflects an involuntarily persistent activation of interacting neural networks involved in experiential consolidation.


The initial appearance of the cranial nerves and related neuronal migration in staged human embryos. The initial development of the cranial nerves was studied in 245 human embryos of stages 10-23 (4-8 postfertilizational weeks). Significant findings in the human embryo include the following. (1) Neuronal migration is a characteristic feature in the development of all the cranial nerves at stages 13-18, with the exception of the somatic efferent group. (2) The somatic efferent and the visceral efferent neurons are arranged respectively in ventrolateral and ventromedial columns (stages 13-17). (3) The ventrolateral column gives rise to somatic efferent nuclei; the neurons of the hypoglossal nerve develop rapidly and show a segmental organization as four roots that innervate three of the four occipital somites (stage 13); the abducent nucleus becomes displaced rostrally by a change in the rhombomeric pattern at stage 16. (4) The ventromedial column, originally continuous in rhombomeres 2-7, gives rise to visceral efferent and pharyngeal efferent nuclei. (5) All the 'true' cranial nerves (III-XII) are recognizable by stage 16. (6) In a primary migration the visceral efferent neurons proceed mediolaterally and accumulate dorsolaterally as nuclei (stages 13, 14); they differentiate into salivatory nuclei (stages 16, 17). (7) A secondary migration involves the pharyngeal efferent neurons (of nerves V and IX-XI), which also proceed mediolaterally and then form ventrolateral nuclei (stages 17, 18). (8) The facial complex shows a distinctive development in that its neural crest arises from the lateral wall of the neural folds/tube. Moreover, the migration of its pharyngeal efferent neurons is delayed, which may be related to the formation of the internal genu, and the motor nucleus begins to appear only at stage 23. (9) The sequence of appearance of afferent constituents is: cranial ganglia (stage 12), mesencephalic trigeminal nucleus (stage 15), vestibular nuclei (stages 18-22), and cochlear nuclei (stage 19). The unsatisfactory term special is avoided and the term pharyngeal for air-breathing vertebrates replaces branchial. The six functional categories used here are vestibulocochlear, somatic afferent, visceral afferent, visceral efferent, pharyngeal efferent, and somatic efferent, together with appropriate abbreviations. The cardiac and hypoglossal neural crests are included, and it is emphasized that all the ectodermal placodes develop within the 'ectodermal ring'.