Study protocol for a pragmatic randomised controlled trial evaluating efficacy of a smoking cessation e-'Tabac Info Service': ee-TIS trial.
A French national smoking cessation service, Tabac Info Service, has been developed to provide an adapted quitline and a web and mobile application involving personalised contacts (eg, questionnaires, advice, activities, messages) to support smoking cessation. This paper presents the study protocol of the evaluation of the application (e-intervention Tabac Info Service (e-TIS)). The primary objective is to assess the efficacy of e-TIS. The secondary objectives are to (1) describe efficacy variations with regard to users' characteristics, (2) analyse mechanisms and contextual conditions of e-TIS efficacy. The study design is a two-arm pragmatic randomised controlled trial including a process evaluation with at least 3000 participants randomised to the intervention or to the control arm (current practices). Inclusion criteria are: aged 18 years or over, current smoker, having completed the online consent forms, possessing a mobile phone with android or apple systems and using mobile applications, wanting to stop smoking sooner or later. The primary outcome is the point prevalence abstinence of 7 days at 6 months later. Data will be analysed in intention to treat (primary) and per protocol analyses. A logistic regression will be carried out to estimate an OR (95% CI) for efficacy. A multivariate multilevel analysis will explore the influence on results of patients' characteristics (sex, age, education and socioprofessional levels, dependency, motivation, quit experiences) and contextual factors, conditions of use, behaviour change techniques. The study protocol was reviewed by the ethical and deontological institutional review board of the French Institute for Public Health Surveillance on 18 April 2016. The findings of this study will allow us to characterise the efficacy of e-TIS and conditions of its efficacy. These findings will be disseminated through peer-reviewed articles. NCT02841683; Pre-results.


The Force of Numbers: Investigating Manual Signatures of Embodied Number Processing.
The study has two objectives: (1) to introduce grip force recording as a new technique for studying embodied numerical processing; and (2) to demonstrate how three competing accounts of numerical magnitude representation can be tested by using this new technique: the Mental Number Line (MNL), A Theory of Magnitude (ATOM) and Embodied Cognition (finger counting-based) account. While 26 healthy adults processed visually presented single digits in a go/no-go n-back paradigm, their passive holding forces for two small sensors were recorded in both hands. Spontaneous and unconscious grip force changes related to number magnitude occurred in the left hand already 100-140 ms after stimulus presentation and continued systematically. Our results support a two-step model of number processing where an initial stage is related to the automatic activation of all stimulus properties whereas a later stage consists of deeper conscious processing of the stimulus. This interpretation generalizes previous work with linguistic stimuli and elaborates the timeline of embodied cognition. We hope that the use of grip force recording will advance the field of numerical cognition research.


Research highlights: cell separation at the bench and beyond.
We highlight recent progress in applying micro- and nanotechnology enabled cell separations to life sciences and clinical use. Microfluidic systems operate on a scale that matches that of cells (10-100 μm) and therefore allow interfacing and separations that are sensitive at this scale. Given the corresponding dimensions, it is not surprising that a wide array of microfluidic cell separation technologies have been developed using hydrodynamic, electrical, magnetic and optical forces, and have been applied to a range of biological and clinical problems in sample preparation. Passive separation approaches have distinct advantages for point of care applications or when downstream cell-based therapies are envisioned. We highlight a recent approach that allows for passive hydrodynamic filtering of cells over almost two orders of magnitude in flow conditions, which allowed the researchers to interface with a standard manual pipettor, creating a "microfluidic pipette tip". In a second work, passive separation by size yields distinct populations of mesenchymal stem cells that can be used therapeutically. The researchers report on other biophysical separations that would be expected to refine these cell populations further for the most efficacious cell-based therapies. In an intriguing twist, we highlight a creative idea in which stem cell populations could potentially also be extracted from a patient with less invasive surgeries, performing the separation using magnetic nanoparticles in vivo without bulk tissue disruption. New cell separation technologies will continue to be demonstrated, however, a major research thrust appears to be now developing these technologies to address unique application niches in point-of-care sample preparation for research and diagnostics or cell-based therapies.


Pro-inflammatory cytokines: Useful markers for the diagnosis of canine mammary tumours?
The aim of the present study was to analyse the expression of 60 pro-inflammatory cytokines as possible markers of malignancy in canine mammary tumours using a human cytokine antibody array. The cytokines were grouped into two different categories: (1) cytokines in which expression indicated the presence of a mammary tumour and (2) cytokines in which expression differentiated between simple mammary adenoma, tubulopapillary carcinoma or complex carcinoma. These data suggest that specific pro-inflammatory cytokines could be useful as tools for the diagnosis of canine mammary tumours.


Bimodal analysis reveals a general scaling law governing nondirected and chemotactic cell motility.
Cell motility is a fundamental process with relevance to embryonic development, immune response, and metastasis. Cells move either spontaneously, in a nondirected fashion, or in response to chemotactic signals, in a directed fashion. Even though they are often studied separately, both forms of motility share many complex processes at the molecular and subcellular scale, e.g., orchestrated cytoskeletal rearrangements and polarization. In addition, at the cellular level both types of motility include persistent runs interspersed with reorientation pauses. Because there is a great range of variability in motility among different cell types, a key challenge in the field is to integrate these multiscale processes into a coherent framework. We analyzed the motility of Dictyostelium cells with bimodal analysis, a method that compares time spent in persistent versus reorientation mode. Unexpectedly, we found that reorientation time is coupled with persistent time in an inverse correlation and, surprisingly, the inverse correlation holds for both nondirected and chemotactic motility, so that the full range of Dictyostelium motility can be described by a single scaling relationship. Additionally, we found an identical scaling relationship for three human cell lines, indicating that the coupling of reorientation and persistence holds across species and making it possible to describe the complexity of cell motility in a surprisingly general and simple manner. With this new perspective, we analyzed the motility of Dictyostelium mutants, and found four in which the coupling between two modes was altered. Our results point to a fundamental underlying principle, described by a simple scaling law, unifying mechanisms of eukaryotic cell motility at several scales.


Target international normalized ratio values for preventing thromboembolic and hemorrhagic events in Japanese patients with non-valvular atrial fibrillation: results of the J-RHYTHM Registry.
Target anticoagulation levels for warfarin in Japanese patients with non-valvular atrial fibrillation (NVAF) are unclear. Of 7,527 patients with NVAF, 1,002 did not receive warfarin (non-warfarin group), and the remaining patients receiving warfarin were divided into 5 groups based on their baseline international normalized ratio (INR) of prothrombin time (≤1.59, 1.6-1.99, 2.0-2.59, 2.6-2.99, and ≥3.0). Patients were followed-up prospectively for 2 years. Primary endpoints were thromboembolic events (cerebral infarction, transient ischemic attack, and systemic embolism), and major hemorrhage requiring hospital admission. During the follow-up period, thromboembolic events occurred in 3.0% of non-warfarin group, but at lower frequencies in the warfarin groups (2.0, 1.3, 1.5, 0.6, and 1.8%/2 years for INR values of ≤1.59, 1.6-1.99, 2.0-2.59, 2.6-2.99, and ≥3.0, respectively; P=0.0059). Major hemorrhage occurred more frequently in warfarin groups (1.5, 1.8, 2.4, 3.3, and 4.1% for INR values ≤1.59, 1.6-1.99, 2.0-2.59, 2.6-2.99, and ≥3.0, respectively; P=0.0041) than in non-warfarin group (0.8%/2 years). These trends were maintained when the analyses were confined to patients aged ≥70 years. An INR of 1.6-2.6 is safe and effective at preventing thromboembolic events in patients with NVAF, particularly patients aged ≥70 years. An INR of 2.6-2.99 is also effective, but associated with a slightly increased risk in major hemorrhage. (UMIN Clinical Trials Registry UMIN000001569)


Morphological changes in taste buds of the fungiform papillae after a single dose of x-ray irradiation in rats.
The qualitative and quantitative changes occurring in the taste buds of the fungiform papillae of male Sprague-Dawley rats (aged 51-125 days old) after x-ray irradiation were studied. A single dose of 2000 Roentgen (R) was delivered and observations were made at 0, 3, 7, 14, 21 and 30 days after x-ray irradiation. The changes in taste bud morphology were interpreted first as degenerative and then as regenerative at various stages of the study. The degenerative process appeared at about the third day and reached a peak at 7 days. Regeneration then began after a further week and the taste buds appeared mature at about 30 days. While the number of fungiform papillae remained constant throughout the study, there was a marked loss of taste buds on these papillae at 7 days. At 14 days, the number of taste buds increased from this low level, and by 30 days the number approached the control values. Initial taste bud loss could be attributed to a direct irradiation damage leading to taste bud cell desquamation combined with a concomitant mitotic inhibition of the stratum germinativum.


Prevention of severe acute kidney injury by implementation of care bundles: Some progress but still a lot of work ahead.
Routine health data, dating from 1990 to 2015, show a dramatic rise in the incidence of acute kidney injury (AKI) in the general population and an apparent increase in acute dialysis (AKI-D) prompting calls for improvement of care in AKI patients. Recent reports suggest flattening of population-based AKI occurrence rates or declining frequencies of AKI or AKI-D in subgroups of high-risk patients with known onset of AKI. These temporal changes in AKI occurrence relate at least in part to the widespread implementation of AKI prevention care bundles. Nevertheless, AKI frequencies and AKI morbidity and mortality remain high. Incorporation of real-time electronic alerts and the use of novel biomarkers in high-risk patients or patients with known onset of AKI may result in further reductions of AKI incidences by early recognition of AKI and by timely conservative management of AKI. There is an urgent need for further treatment options of established AKI.


Superficial necrolytic dermatitis (necrolytic migratory erythema) in dogs.
Twenty-two dogs with superficial necrolytic dermatitis were evaluated prospectively, twenty-one of which had characteristic crusting lesions of the paw pads. Histologically, epidermal lesions included parakeratosis and laminar intracellular edema. The plasma amino acid concentrations of eight dogs were markedly depressed. Nine dogs had terminal diabetes mellitus. These clinical and morphologic findings were strikingly similar to those of necrolytic migratory erythema in human beings, the most common cause of which is hyperglucagonemia due to islet cell tumor of the pancreas. No pancreatic tumors were found in these dogs; plasma glucagon concentrations in the five dogs tested were normal. The serum alkaline phosphatase concentrations were elevated in all dogs. Severe vacuolar hepatopathy, suggesting metabolically or hormonally induced hepatic dysfunction, was found in 21 dogs at necropsy or by biopsy; one dog had ultrasonographic abnormalities of the liver. Histopathologically, severe vacuolar alteration resulted in parenchymal collapse and nodular regeneration, which grossly mimicked cirrhosis. Although the definitive metabolic stimulus was not discovered for the cutaneous and hepatic lesions, the similarity of the cutaneous and biochemical features of canine superficial necrolytic dermatitis to human necrolytic migratory erythema warrants further investigation into possible underlying pancreatic hormonal dysfunction.


Cytotoxic-T-lymphocyte-mediated cytolysis of L cells persistently infected with Chlamydia spp.
Persistent chlamydial infections have been proposed as a means whereby chlamydiae evade immune resolution of infection. Such a mechanism would require evasion not only of the humoral immune responses but also of cell-mediated immune responses. We hypothesized that if such a mechanism is important, persistently infected cells should not be recognized by cytotoxic T cells. Persistent infections were simulated in vitro by treatment of Chlamydia trachomatis- or Chlamydia psittaci-infected cells with gamma interferon (IFN-gamma), penicillin, or tryptophan depletion. Cultures were examined for induction of a chlamydial stress response (measured by transcription of groesl RNA) and for the effects on viability, infectivity, morphology, and immune recognition. Although both IFN-gamma and penicillin induced aberrant chlamydial morphology and growth, we did not find evidence that these treatments elicited a classical stress response. In addition, T-cell-mediated lysis of Chlamydia-infected target cells treated with IFN-gamma or penicillin or grown in tryptophan-deficient media was examined. The immune cell-mediated lysis of these treated infected cells demonstrated that despite the effects of these compounds on chlamydial growth and development, the infected cells continued to be efficiently recognized and killed by cytotoxic T cells. Thus, it seems unlikely that these in vitro models of persistence represent functional mechanisms to evade immune clearance.


Repression of Igf1 expression by Ezh2 prevents basal cell differentiation in the developing lung.
Epigenetic mechanisms involved in the establishment of lung epithelial cell lineage identities during development are largely unknown. Here, we explored the role of the histone methyltransferase Ezh2 during lung lineage determination. Loss of Ezh2 in the lung epithelium leads to defective lung formation and perinatal mortality. We show that Ezh2 is crucial for airway lineage specification and alveolarization. Using optical projection tomography imaging, we found that branching morphogenesis is affected in Ezh2 conditional knockout mice and the remaining bronchioles are abnormal, lacking terminally differentiated secretory club cells. Remarkably, RNA-seq analysis revealed the upregulation of basal genes in Ezh2-deficient epithelium. Three-dimensional imaging for keratin 5 further showed the unexpected presence of a layer of basal cells from the proximal airways to the distal bronchioles in E16.5 embryos. ChIP-seq analysis indicated the presence of Ezh2-mediated repressive marks on the genomic loci of some but not all basal genes, suggesting an indirect mechanism of action of Ezh2. We found that loss of Ezh2 de-represses insulin-like growth factor 1 (Igf1) expression and that modulation of IGF1 signaling ex vivo in wild-type lungs could induce basal cell differentiation. Altogether, our work reveals an unexpected role for Ezh2 in controlling basal cell fate determination in the embryonic lung endoderm, mediated in part by repression of Igf1 expression.


The maize Dwarf3 gene encodes a cytochrome P450-mediated early step in Gibberellin biosynthesis.
Gibberellins (GAs) are phytohormones required for normal growth and development in higher plants. The Dwarf3 (D3) gene of maize encodes an early step in the GA biosynthesis pathway. We transposon-tagged the D3 gene using Robertson's Mutator (Mu) and showed that the mutant allele d3.2::Mu8 is linked to a Mu8 element. The DNA flanking the Mu8 element was cloned and shown to be linked to the d3 locus by mapping in a high-resolution population developed by selecting for recombination between d3 and linked genetic markers. To establish unambiguously the identity of the cloned gene as D3, a second mutant allele of D3 (d3.4) was also cloned and characterized using the d3.2::Mu8 sequences as a probe. d3.4 was found to have a novel insertion element, named Sleepy, inserted into an exon. A third mutant allele, d3.1, which has the same size 3' restriction fragments as d3.4 but different 5' restriction fragments, was found to contain a Sleepy insertion at the same position as d3.4. On the basis of the pedigree, Sleepy insertion, and restriction map, d3.1 appears to represent a recombinational derivative of d3.4. The D3 gene encodes a predicted protein with significant sequence similarity to cytochrome P450 enzymes. Analysis of D3 mRNA showed that the D3 transcript is expressed in roots, developing leaves, the vegetative meristem, and suspension culture cells. We detected reduced D3 mRNA levels in the mutant allele d3.5.


Homocysteine levels in the vitreous of proliferative diabetic retinopathy and rhegmatogenous retinal detachment: its modulating role on lysyl oxidase.
Homocysteine (Hcys), a well-known inducer of vascular endothelial cell damage has been associated with extracellular matrix changes. Lysyl oxidase (LOX) is a copper-dependent amine oxidase that initiates the covalent cross-linking of collagen and elastin in the extracellular matrix (ECM). LOX contributes to the structural integrity of the ECM, and low LOX activity could promote ECM disorganization. Hydroxyproline levels are used to predict collagen turnover status, and most of the endogenous hydroxyproline present in biological fluids is derived from the degradation of various forms of collagen. As Hcys is known to regulate ECM turnover and also inhibit LOX activity, the purpose of this study was to estimate the vitreous levels of Hcys in eyes with proliferative diabetic retinopathy (PDR) and rhegmatogenous retinal detachment (RRD) and to correlate the effect of Hcys, if any on LOX activity. Undiluted human vitreous specimens obtained during vitreoretinal surgeries for PDR (n = 18) and RRD (n = 17) were used. Vitreous specimens from donor eyeballs were used as control (n = 19). Hcys was estimated by HPLC using a fluorescent detector. Hydroxyproline was estimated spectrophotometrically. The total vitreous Hcys level was found to be increased significantly in PDR (P = 0.011) and in RRD (P = 0.001) compared with that in control samples. Hydroxyproline was significantly increased in PDR (P = 0.049) and RRD (P = 0.007) compared with the level in control samples. There was a significant negative correlation between the Hcys level and the specific activity of LOX in PDR (P = 0.040) and in RRD (P = 0.029) This report shows that increased vitreous Hcys in PDR and RRD is associated with a significant decrease in LOX-specific activity along with an increase in collagen turnover.


Contingent Valuation: A Pilot Study for Eliciting Willingness to Pay for a Reduction in Mortality From Vaccine-Preventable Illnesses for Children and Adults in Bangladesh.
The contingent valuation (CV) method elicits willingness to pay (WTP) for calculating the value of statistical life (VSL). CV approaches for assessing VSL are uncommon in many low and middle-income countries (LMICs). Between 2008 and 2018 only 44 articles utilized WTP in a health-related field and of these only 5 (11%) utilized CV to assess the WTP for a mortality risk reduction. We elicit WTP estimates and compute VSL using the CV method in Bangladesh. The pilot study was primarily aimed at developing best practice guidelines for CV studies in LMICs to get more robust WTP estimates. To this end, we explored three methodological a) Varying the name of the intervention, keeping all other characteristics constant; b) varying the effectiveness of the health intervention and c) offering an overnight period to think about the WTP scenario. The survey was administered 413 randomly selected participants. VSL was for a 1/3000 mortality risk reduction. We had more males (54%) than females (46%) and the mean annual self-reported income was $5,683.36. Mean VSL is $11,339.70 with a median of $10,413. The ratio of child: adult WTP is approximately 1 by both gender and age category. The vaccine intervention had the largest amount of $0 WTP and protest responses (52% and 58% respectively). 93% of the participants were able to describe (teach-back) the vaccine effectiveness using their own family as an example. Our study provides empirical evidence on how to better generate CV surveys to produce more robust WTP estimates.


A high-conductance chemo-optogenetic system based on the vertebrate channel Trpa1b.
Optogenetics is a powerful research approach that allows localized optical modulation of selected cells within an animal via the expression of genetically encoded photo-excitable ion channels. Commonly used optogenetic techniques rely on the expression of microbial opsin variants, which have many excellent features but suffer from various degrees of blue spectral overlap and limited channel conductance. Here, we expand the optogenetics toolbox in the form of a tunable, high-conductance vertebrate cation channel, zTrpa1b, coupled with photo-activated channel ligands, such as optovin and 4g6. Our results demonstrate that zTrpa1b/ligand pairing offers high light sensitivity, millisecond-scale response latency in vivo, as well as adjustable channel off latency. Exogenous in vivo expression of zTrpa1b in sensory neurons allowed subcellular photo-activation, enabling light-dependent motor control. zTrpa1b/ligand was also suitable for cardiomyocyte pacing, as shown in experiments performed on zebrafish hearts in vivo as well as in human stem cell-derived cardiomyocytes in vitro. Therefore, zTrpa1b/optovin represents a novel tool for flexible, high-conductance optogenetics.


Porous-carbon-based Mo2C nanocomposites as excellent microwave absorber: a new exploration.
In this study, we report a porous-carbon-based Mo2C nanocomposite (NCs) as the microwave absorber via typical carbothermal reduction using metal-organic frameworks (MOFs) and polyoxometalates (POMs) as the precursors, which have been rarely applied in electromagnetic (EM) wave absorption areas. The elaborately designed NCs not only bring about good impedance matching, but also possess strong dissipation ability due to the large surface areas and porous features. Thanks to the material characteristics as well as structural advantages, the as-prepared Mo2C/C NCs with 20 wt% sample loading exhibit remarkable microwave absorbing performance. The minimum RL value reaches -49.19 dB at matching thickness of 2.6 mm, and the best effective bandwidth (RL < -10 dB) of 4.56 GHz at 1.70 mm was also achieved. Moreover, the NCs overcome the intrinsic drawback of traditional carbon materials, that is, centralized effective absorption always occurred at high frequency (>10 GHz) and the minimum RL value of the NCs shifted to 9.04 GHz. Clearly, in this study, we not only developed the Mo2C NC as the new light absorber, but also paved the way to synthesize other available transition metal carbides using MOFs and POMs.


Worldwide time trends in the prevalence of symptoms of asthma, allergic rhinoconjunctivitis, and eczema in childhood: ISAAC Phases One and Three repeat multicountry cross-sectional surveys.
Data for trends in prevalence of asthma, allergic rhinoconjunctivitis, and eczema over time are scarce. We repeated the International Study of Asthma and Allergies in Childhood (ISAAC) at least 5 years after Phase One, to examine changes in the prevalence of symptoms of these disorders. For the ISAAC Phase Three study, between 2002 and 2003, we did a cross-sectional questionnaire survey of 193,404 children aged 6-7 years from 66 centres in 37 countries, and 304,679 children aged 13-14 years from 106 centres in 56 countries, chosen from a random sample of schools in a defined geographical area. Phase Three was completed a mean of 7 years after Phase One. Most centres showed a change in prevalence of 1 or more SE for at least one disorder, with increases being twice as common as decreases, and increases being more common in the 6-7 year age-group than in the 13-14 year age-group, and at most levels of mean prevalence. An exception was asthma symptoms in the older age-group, in which decreases were more common at high prevalence. For both age-groups, more centres showed increases in all three disorders more often than showing decreases, but most centres had mixed changes. The rise in prevalence of symptoms in many centres is concerning, but the absence of increases in prevalence of asthma symptoms for centres with existing high prevalence in the older age-group is reassuring. The divergent trends in prevalence of symptoms of allergic diseases form the basis for further research into the causes of such disorders.


Use of Biomarker Modulation in Normal Mammary Epithelium as a Correlate for Efficacy of Chemopreventive Agents Against Chemically Induced Cancers.
In both estrogen receptor/progesterone receptor-positive (ER+/PR+) human breast cancer and in ER+/PR+ cancers in the methylnitrosourea (MNU)-induced rat model, short-term modulation of proliferation in early cancers predicts preventive/therapeutic efficacy. We determined the effects of known effective/ineffective chemopreventive agents on proliferative index (PI) in both rat mammary epithelium and small cancers. Female Sprague-Dawley rats were treated with MNU at 50 days of age. Five days later, the rats were treated with the individual compounds for a period of 14 days. At that time, normal mammary tissue from the inguinal gland area was surgically removed. After removal, the rats remained on the agents for an additional 5 months. This cancer prevention study confirmed our prior results of striking efficacy with tamoxifen, vorozole, Targretin, and gefitinib, and no efficacy with metformin, naproxen, and Lipitor. Employing a separate group of rats, the effects of short-term (7 days) drug exposure on small palpable cancers were examined. The PI in both small mammary cancers and in normal epithelium from control rats was >12%. In agreement with the cancer multiplicity data, tamoxifen, vorozole, gefitinib, and Targretin all strongly inhibited proliferation (>65%; P < 0.025) in the normal mammary epithelium. The ineffective agents metformin, naproxen, and Lipitor minimally affected PI. In the small cancers, tamoxifen, vorozole, and Targretin all reduced the PI, while metformin and Lipitor failed to do so. Thus, short-term changes in the PI in either normal mammary epithelium or small cancers correlated with long-term preventive efficacy in the MNU-induced rat model.


Somatization among older primary care attenders.
The importance of somatization among older primary care attenders is unclear. We aimed to establish the prevalence, persistence and associations of somatization among older primary care attenders, and the associations of frequent attendance. One hundred and forty primary care attenders over 65 years were rated twice, 10 months apart, on measures of somatization, psychiatric status, physical health and attendance. The syndrome of GMS hypochondriacal neurosis had a prevalence of 5% but was transient. Somatized symptoms and attributions were persistent and associated with depression, physical illness and perceived poor social support. Frequent attenders (top third) had higher rates of depression, physical illness and somatic symptoms, and lower perceived support. Somatization is common among older primary care attenders and has similar correlates to younger primary care somatizers. Psychological distress among older primary care attenders is associated with frequent attendance. Improved recognition should result in benefits to patients and services.


Validation of the Asthma Illness Representation Scale (AIRS).
Research has suggested a link between parents' illness representations (IRs), use of complementary and alternative medicine, inhaled/oral corticosteroids and leukotriene antagonists, and children's health outcomes. The Asthma Illness Representation Scale (AIRS) provides a structured assessment of the key components of asthma IRs allowing the healthcare provider (HCP) to quickly identify areas of discordance with the professional model of asthma management. These analyses extend the initial validation of the AIRS and compares data from the original study conducted among a primarily white and African American sample in Rochester, NY (N = 228) with data obtained from a predominantly inner-city, ethnic minority sample (Puerto Rican, African American, and Afro-Caribbean) from the Bronx, New York (N = 109). A larger proportion of the Rochester sample was white and non-poor and had graduated high school. Bronx parents were more likely to perceive their child's asthma to be moderate or severe than the Rochester parents. Bronx children were older and had longer duration of asthma and reported more acute health care visits (past year). Bronx parents reported total AIRS scores more closely aligned with the lay model than Rochester parents. The AIRS instrument demonstrated acceptable internal reliability among the Bronx sample (total score alpha = 0.82) and the AIRS subscale Cronbach's alpha coefficients were remarkably similar to those obtained from the original validation study (range = 0.54-0.83). Poor parents and those with less than a high school education had lower total AIRS scores than their counterparts. White parents had AIRS scores more closely aligned with the professional model compared to each of the ethnic subgroups. A perception of less severe asthma, fewer reports of asthma and somatization symptoms, and a positive HCP relationship were associated with IRs congruent with the professional model. IRs aligned with the professional model were associated with fewer acute asthma-related healthcare visits. The AIRS instrument exhibited good internal reliability, external validity, and differentiated parents based on ethnicity, poverty, and education. Assessment of asthma IRs during the healthcare visit will allow the HCP and parent to discuss and negotiate a shared asthma management plan for the child, which will hopefully lead to improved medication adherence and asthma health outcomes.


Templated blue phases.
Cholesteric blue phases of a chiral liquid crystal are interesting examples of self-organised three-dimensional nanostructures formed by soft matter. Recently it was demonstrated that a polymer matrix introduced by photopolymerization inside a bulk blue phase not only stabilises the host blue phase significantly, but also serves as a template for blue phase ordering. We show with numerical modelling that the transfer of the orientational order of the blue phase to the surfaces of the polymer matrix, together with the resulting surface anchoring, can account for the templating behaviour of the polymer matrix inducing the blue phase ordering of an achiral nematic liquid crystal. Furthermore, tailoring the anchoring conditions of the polymer matrix surfaces can bring about orientational ordering different from those of bulk blue phases, including an intertwined complex of the polymer matrix and topological line defects of orientational order. Optical Kerr response of templated blue phases is explored, finding large Kerr constants in the range of K = 2-10 × 10(-9) m V(-2) and notable dependence on the surface anchoring strength. More generally, the presented numerical approach is aimed to clarify the role and actions of templating polymer matrices in complex chiral nematic fluids, and further to help design novel template-based materials from chiral liquid crystals.


Tooth Agenesis in Patients with Complete Bilateral Cleft Lip and Palate.
The aim of this study was to compare the number of teeth in the premaxilla (cleft area) and to assess the presence of tooth agenesis with the ultimate goal of finding whether differences existed in the two suggested subtypes of nonsyndromic bilateral cleft lip and palate (BCLP) patients. A retrospective comparative study was conducted, where cone- beam computed tomography (CBCT) images of 22 BCLP patients (10 males and 12 females, with age range 8 to 14, mean age 9.6 years) obtained from archive of Cleft Clinic, affiliated to the Oral and Maxillofacial Surgery department, Ain-Shams University, Cairo, Egypt were assessed. CBCT images were divided into two subtypes: Group (P) - this represents BCLP characterized by well-developed (P) prominent premaxilla and SNA >80 ± 2 and Group (R) - this represents BCLP characterized by ill-developed (R) rudimentary premaxilla and SNA <80 ± 2. The number of teeth in the premaxilla was evaluated, and the presence of agenesis in the premaxilla and posterior segments was documented. Unpaired-Student's t-test was used to compare the number of teeth among the two groups and to test significance at P < 0.05. Intraobserver agreement was assessed using alpha (Cronbach) reliability analysis. A descriptive analysis using percentages was performed to characterize tooth agenesis. A significant difference was found in the number of teeth in the premaxilla among Group R and Group P. Maxillary lateral incisor showed the highest percentage of agenesis in the two groups followed by the maxillary second premolar. The number of teeth might be useful in differentiating different subtypes of nonsyndromic BCLP.


Emergency physician awareness of prehospital procedures and medications.
Maintaining patient safety during transition from prehospital to emergency department (ED) care depends on effective handoff communication between providers. We sought to determine emergency physicians' (EP) knowledge of the care provided by paramedics in terms of both procedures and medications, and whether the use of a verbal report improved physician accuracy. We conducted a 2-phase observational survey of a convenience sample of EPs in an urban, academic ED. In this large ED paramedics have no direct contact with physicians for non-critical patients, giving their report instead to the triage nurse. In Phase 1, paramedics gave verbal report to the triage nurse only. In Phase 2, a research assistant (RA) stationed in triage listened to this report and then repeated it back verbatim to the EPs caring for the patient. The RA then queried the EPs 90 minutes later regarding their patients' prehospital procedures and medications. We compared the accuracy of these 2 reporting methods. There were 163 surveys completed in Phase 1 and 116 in Phase 2. The oral report had no effect on EP awareness that the patient had been brought in by ambulance (86% in Phase 1 and 85% in Phase 2.) The oral report did improve EP awareness of prehospital procedures, from 16% in Phase 1 to 45% in Phase 2, OR=4.28 (2.5-7.5). EPs were able to correctly identify all oral medications in 18% of Phase 1 cases and 47% of Phase 2 cases, and all IV medications in 42% of Phase 1 cases and 50% of Phase 2 cases. The verbal report led to a mild improvement in physician awareness of oral medications given, OR=4.0 (1.09-14.5), and no improvement in physician awareness of IV medications given, OR=1.33 (0.15-11.35). Using a composite score of procedures plus oral plus IV medications, physicians had all three categories correct in 15% of Phase 1 and 39% of Phase 2 cases (p<0.0001). EPs in our ED were unaware of many prehospital procedures and medications regardless of the method used to provide this information. The addition of a verbal hand-off report resulted in a modest improvement in overall accuracy.


Evaluation of the antiangiogenic potential of AQ4N.
A number of cytotoxic chemotherapy agents tested at low concentrations show antiangiogenic properties with limited cytotoxicity, e.g., cyclophosphamide, tirapazamine, and mitoxantrone. AQ4N is a bioreductive alkylaminoanthraquinone that is cytotoxic when reduced to AQ4; hence, it can be used to target hypoxic tumor cells. AQ4N is structurally similar to mitoxantrone and was evaluated for antiangiogenic properties without the need for bioreduction. The effect of AQ4N and fumagillin on human microvascular endothelial cells (HMEC-1) was measured using a variety of in vitro assays, i.e., 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide, wound scrape, tubule formation, rat aortic ring, and invasion assays. Low-dose AQ4N (20 mg/kg) was also given in vivo to mice bearing a tumor in a dorsal skin flap. AQ4N (10(-11) to 10(-5) mol/L) had no effect on HMEC-1 viability. AQ4N (10(-9) to 10(-5)mol/L) caused a sigmoidal dose-dependent inhibition of endothelial cell migration in the wound scrape model. Fumagillin showed a similar response over a lower dose range (10(-13) to 10(-9) mol/L); however, the maximal inhibition was less (25% versus 43% for AQ4N). AQ4N inhibited HMEC-1 cell contacts on Matrigel (10(-8) to 10(-5) mol/L), HMEC-1 cell invasion, and sprouting in rat aorta explants. Immunofluorescence staining with tubulin, vimentim, dynein, and phalloidin revealed that AQ4N caused disruption to the cell cytoskeleton. When AQ4N (20 mg/kg) was given in vivo for 5 days, microvessels disappeared in LNCaP tumors grown in a dorsal skin flap. This combination of assays has shown that AQ4N possesses antiangiogenic effects in normoxic conditions, which could potentially contribute to antitumor activity.


The 'Fat Mass and Obesity Related' (FTO) gene: Mechanisms of Impact on Obesity and Energy Balance.
A cluster of single nucleotide polymorphisms (SNPs) in the first intron of the fat mass and obesity related (FTO) gene were the first common variants discovered to be associated with body mass index and body fatness. This review summarises what has been later discovered about the biology of FTO drawing together information from both human and animal studies. Subsequent work showed that the 'at risk' alleles of these SNPs are associated with greater food intake and increased hunger/lowered satiety, but are not associated with altered resting energy expenditure or low physical activity in humans. FTO is an FE (II) and 2-oxoglutarate dependent DNA/RNA methylase. Contrasting the impact of the SNPs on energy balance in humans, knocking out or reducing activity of the Fto gene in the mouse resulted in lowered adiposity, elevated energy expenditure with no impact on food intake (but the impact on expenditure is disputed). In contrast, overexpression of the gene in mice led to elevated food intake and adiposity, with no impact on expenditure. In rodents, the Fto gene is widely expressed in the brain including hypothalamic nuclei linked to food intake regulation. Since its activity is 2-oxoglutarate dependent it could potentially act as a sensor of citrate acid cycle flux, but this function has been dismissed, and instead it has been suggested to be much more likely to act as an amino acid sensor, linking circulating AAs to the mammalian target of rapamycin complex 1. This may be fundamental to its role in development but the link to obesity is less clear. It has been recently suggested that although the obesity related SNPs reside in the first intron of FTO, they may not only impact FTO but mediate their obesity effects via nearby genes (notably RPGRIP1L and IRX3).


Pulsatile and sustained gonadotropin-releasing hormone (GnRH) receptor signaling: does the Ca2+/NFAT signaling pathway decode GnRH pulse frequency?
Gonadotropin-releasing hormone (GnRH) acts via 7 transmembrane region receptors on gonadotrophs to stimulate synthesis and secretion of the luteinizing hormone and follicle-stimulating hormone. It is secreted in pulses, and its effects depend on pulse frequency, but decoding mechanisms are unknown. Here we have used (nuclear factor of activated T-cells 2 (NFAT2)-emerald fluorescent protein) to monitor GnRH signaling. Increasing [Ca(2+)](i) causes calmodulin/calcineurin-dependent nuclear NFAT translocation, a response involving proteins (calmodulins and NFATs) that decode frequency in other systems. Using live cell imaging, pulsatile GnRH caused dose- and frequency-dependent increases in nuclear NFAT2-emerald fluorescent protein, and at low frequency, translocation simply tracked GnRH exposure (albeit with slower kinetics). At high frequency (30-min intervals), failure to return to basal conditions before repeat stimulation caused integrative tracking, illustrating how the relative dynamics of up- and downstream signals can increase efficiency of GnRH action. Mathematical modeling predicted desensitization of GnRH effects on [Ca(2+)](i) and that desensitization would increase with dose, frequency, and receptor number, but no such desensitization was seen in HeLa and/or LbetaT2 cells possibly because pulsatile GnRH did not reduce receptor expression (measured by immunofluorescence). GnRH also caused dose- and frequency-dependent activation of alphaGSU, luteinizing hormone beta, and follicle-stimulating hormone beta luciferase reporters, effects that were blocked by calcineurin inhibition. Pulsatile GnRH also activated an NFAT-responsive luciferase reporter, but this response was directly related to cumulative pulse duration. This together with the lack of desensitization of translocation responses suggests that NFAT may mediate GnRH action but is not a genuine decoder of GnRH pulse frequency.


Regulation of NKB pathways and their roles in the control of Kiss1 neurons in the arcuate nucleus of the male mouse.
Kisspeptin (Kiss1) and neurokinin B (NKB) (encoded by the Kiss1 and Tac2 genes, respectively) are indispensable for reproduction. In the female of many species, Kiss1 neurons in the arcuate nucleus (ARC) coexpress dynorphin A and NKB. Such cells have been termed Kiss1/NKB/Dynorphin (KNDy) neurons, which are thought to mediate the negative feedback regulation of GnRH/LH secretion by 17β-estradiol. However, we have less knowledge about the molecular physiology and regulation of Kiss1/Kiss1-expressing neurons in the ARC of the male. Our work focused on the adult male mouse, where we sought evidence for coexpression of these neuropeptides in cells in the ARC, assessed the role of Kiss1 neurons in negative feedback regulation of GnRH/LH secretion by testosterone (T), and investigated the action of NKB on KNDy and GnRH neurons. Results showed that 1) the mRNA encoding Kiss1, NKB, and dynorphin are coexpressed in neurons located in the ARC; 2) Kiss1 and dynorphin A mRNA are regulated by T through estrogen and androgen receptor-dependent pathways; 3) senktide, an agonist for the NKB receptor (neurokinin 3 receptor, encoded by Tacr3), stimulates gonadotropin secretion; 4) KNDy neurons express Tacr3, whereas GnRH neurons do not; and 5) senktide activates KNDy neurons but has no discernable effect on GnRH neurons. These observations corroborate the putative role for KNDy neurons in mediating the negative feedback effects of T on GnRH/LH secretion and provide evidence that NKB released from KNDy neurons is part of an auto-feedback loop that generates the pulsatile secretion of Kiss1 and GnRH in the male.


The phox homology (PX) domain protein interaction network in yeast.
The phox homology (PX) domain is a phosphoinositide-binding domain that is conserved from yeast to human. Here we show for the first time by genome-wide two-hybrid screens and in vitro binding assays that the PX domain is a bona fide protein interaction domain. The yeast PX domain-only proteins Grd19p (YOR357C) and Ypt35p (YHR105W), as well as the isolated PX domains from Mvp1p (YMR004W), Snx42p/Cvt20p/Atg20p (YDL113C), Vam7p (YGL212W), and Vps17p (YOR132W), yielded a total of 40 reproducible two-hybrid interactions. Thirty-five interactions were found for the full-length proteins of Bem1p (YBR200W), Snx42p, Snx4p/Cvt13p (YJL036W), Vam7p, Vps5p (YOR069W), and Vps17p, but these appear not to require the PX domain, because these interactions could not be reproduced with PX-only baits. Interactions of Grd19p, Vam7p, Vps5p, Vps17p, and Ypt35p with members of the Yip1p family of proteins were detected consistently and were verified by in vitro binding assays. The N-terminal cytoplasmic domain of Yip1p and Yif1p mediates these interactions with PX domains. A mutation in the lipid-binding pocket of Ypt35p that reduces lipid binding markedly does not affect these PX domain protein interactions, arguing that lipid binding uses a different interaction surface than protein binding.


Mexican propolis flavonoids affect photosynthesis and seedling growth.
As a continuous effort to find new natural products with potential herbicide activity, flavonoids acacetin (1), chrysin (2) and 4',7-dimethylnarangenin (3) were isolated from a propolis sample collected in the rural area of Mexico City and their effects on the photosynthesis light reactions and on the growth of Lolium perenne, Echinochloa crus-galli and Physalis ixocarpa seedlings were investigated. Acacetin (1) acted as an uncoupler by enhancing the electron transport under basal and phosphorylating conditions and the Mg(2+)-ATPase. Chrysin (2) at low concentrations behaved as an uncoupler and at concentrations up to 100 μM its behavior was as a Hill reaction inhibitor. Finally, 4',7-dimethylnarangenin (3) in a concentration-dependent manner behaved as a Hill reaction inhibitor. Flavonoids 2 and 3 inhibited the uncoupled photosystem II reaction measured from water to 2,5-dichloro-1,4-benzoquinone (DCBQ), and they did not inhibit the uncoupled partial reactions measured from water to sodium silicomolybdate (SiMo) and from diphenylcarbazide (DPC) to diclorophenol indophenol (DCPIP). These results indicated that chrysin and 4',7-dimethylnarangenin inhibited the acceptor side of PS II. The results were corroborated with fluorescence of chlorophyll a measurements. Flavonoids also showed activity on the growth of seedlings of Lolium perenne and Echinochloa crus-galli.


Disparities in health care are driven by where minority patients seek care: examination of the hospital quality alliance measures.
Racial/ethnic disparities in health care are well documented, but less is known about whether disparities occur within or between hospitals for specific inpatient processes of care. We assessed racial/ethnic disparities using the Hospital Quality Alliance Inpatient Quality of Care Indicators. We performed an observational study using patient-level data for acute myocardial infarction (5 care measures), congestive heart failure (2 measures), community-acquired pneumonia (2 measures), and patient counseling (4 measures). Data were obtained from 123 hospitals reporting to the University HealthSystem Consortium from the third quarter of 2002 to the first quarter of 2005. A total of 320,970 patients 18 years or older were eligible for at least 1 of the 13 measures. There were consistent unadjusted differences between minority and nonminority patients in the quality of care across 8 of 13 quality measures (from 4.63 and 4.55 percentage points for angiotensin-converting enzyme inhibitors for acute myocardial infarction and congestive heart failure [P<.01] to 14.58 percentage points for smoking cessation counseling for pneumonia [P=.02]). Disparities were most pronounced for counseling measures. In multivariate models adjusted for individual patient characteristics and hospital effect, the magnitude of the disparities decreased substantially, yet remained significant for 3 of the 4 counseling measures; acute myocardial infarction (unadjusted, 9.00 [P<.001]; adjusted, 3.82 [P<.01]), congestive heart failure (unadjusted, 8.45 [P=.02]; adjusted, 3.54 [P=.02]), and community-acquired pneumonia (unadjusted, 14.58 [P=.02]; adjusted, 4.96 [P=.01]). Disparities in clinical process of care measures are largely the result of differences in where minority and nonminority patients seek care. However, disparities in services requiring counseling exist within hospitals after controlling for site of care. Policies to reduce disparities should consider the underlying reasons for the disparities.


An oxygen key to the new superconductors.
First it was the physicists, then the chemists, and most recently the materials scientists and ceramists who have hastily included in their annual meetings symposia on the new high-temperature, ceramic superconductors. Below are briefings from the 1987 Spring Meeting of the Materials Research Society (MRS) that was held in Anaheim, California, from 21 to 24 April, 1 week before the American Ceramics Society's conclave in Pittsburgh. With the initial wave of euphoria now past, the atmosphere in Anaheim was decidedly more professional than that of the now fabled "Woodstock of Physics" that was part of the American Physical Society's March Meeting in New York City only 5 weeks before. Nonetheless, perhaps 1500 materials researchers listened to 69 scheduled papers and several late walk-ons that were crammed into a 2-day symposium. With a martial strictness, cochairs Michael Schlüter of AT&T Bell Laboratories and Donald Gubser of the Naval Research Laboratory kept the talks to the allotted 10 minutes each. Except for an impassioned presentation by Juei-Teng Chen of Wayne State University, who sought to convince listeners that a group there had seen dear signs of superconductivity at 240 K, which is ambient temperature during a cold night on the northern plains, no significant indications of room-temperature superconductivity were reported. The most skeptical view was that of Theodore Geballe of Stanford University, who suggested that some of the unreproducible signs seen in several laboratories could be due to something other than superconductivity, as similar effects disappeared in Stanford samples with repeated cycling between room and liquid-nitrogen temperature. If there was one theme at the symposium, it was that oxygen is the key to the family of rare-earth-based ceramic materials now in hand that remain superconducting up to about 100 K.


Temperature-regulated expression of outer membrane proteins in Shigella flexneri.
Bacteria exist widely in a diversity of natural environments. In order to survive adverse conditions such as nutrient depletion, biochemical and biological disturbances, and high temperature, bacteria have developed a wide variety of coping mechanisms. Temperature is one of the most important factors that can enhance the expression of microbial proteins. This study was conducted to investigate how outer membrane proteins (OMPs) of the bacterium Shigella flexneri respond to stress, especially during fever when the host's body temperature is elevated. OMPs of S. flexneri ATCC 12022 and clinical isolate SH057 were extracted from an overnight culture grown at 37, 38.5, and 40°C. Comparisons of the expressed proteins under the different growth conditions were based on equal numbers of bacterial cells loaded in the SDS-PAGE gels. Separated proteins were stained with Coomassie brilliant blue. Selected proteins showing increased expression at 38.5 and 40°C were characterized by performing MALDI-ToF-ToF. Different degrees of expression were demonstrated for different proteins expressed at 37°C compared to 38.5 and 40°C. The proteins with molecular sizes of 18.4, 25.6, and 57.0 kDa showed increased expression level at increasing temperature and were identified as Dps, WrbA, and PepA, respectively. This study revealed that strains of S. flexneri respond at the proteomic level during stress caused by elevated temperature by decreasing the expression of proteins, maintaining the level of important proteins, or enhancing the levels of proteins presumably involved in survival and virulence.


Identification of breast cancer DNA methylation markers optimized for fine-needle aspiration samples.
Random periareolar fine-needle aspiration (RP-FNA) is increasingly used in trials of breast cancer prevention for biomarker assessments. DNA methylation markers may have value as surrogate endpoint biomarkers, but this requires identification of biologically relevant markers suitable for paucicellular, lymphocyte-contaminated clinical samples. Unbiased whole-genome 5-aza-2'-deoxycytidine (5AZA)-induced gene expression assays, followed by several phases of qualitative and quantitative methylation-specific PCR (MSP) testing, were used to identify novel breast cancer DNA methylation markers optimized for clinical FNA samples. The initial 5AZA experiment identified 453 genes whose expression was potentially regulated by promoter region methylation. Informatics filters excluded 273 genes unlikely to yield useful DNA methylation markers. MSP assays were designed for 271 of the remaining genes and, ultimately, 33 genes were identified that were differentially methylated in clinical breast cancer samples, as compared with benign RP-FNA samples, and never methylated in lymphocytes. A subset of these markers was validated by quantitative multiplex MSP in extended clinical sample sets. Using a novel permutation method for analysis of quantitative methylation data, PSAT1, GNE, CPNE8, and CXCL14 were found to correlate strongly with specific clinical and pathologic features of breast cancer. In general, our approach identified markers methylated in a smaller subpopulation of tumor cells than those identified in published methylation array studies. Clinically relevant DNA methylation markers were identified using a 5AZA-induced gene expression approach. These breast cancer-relevant, FNA-optimized DNA methylation markers may have value as surrogate endpoint biomarkers in RP-FNA studies.


Optimising Cognitive Enhancement: Systematic Assessment of the Effects of tDCS Duration in Older Adults.
Transcranial direct current stimulation (tDCS) has been shown to support cognition and brain function in older adults. However, there is an absence of research specifically designed to determine optimal stimulation protocols, and much of what is known about subtle distinctions in tDCS parameters is based on young adult data. As the first systematic exploration targeting older adults, this study aimed to provide insight into the effects of variations in stimulation duration. Anodal stimulation of 10 and 20 min, as well as a sham-control variant, was administered to dorsolateral prefrontal cortex. Stimulation effects were assessed in relation to a novel attentional control task. Ten minutes of anodal stimulation significantly improved task-switching speed from baseline, contrary to the sham-control and 20 min variants. The findings represent a crucial step forwards for methods development, and the refinement of stimulation to enhance executive function in the ageing population.


Choosing family medicine. What influences medical students?
To explore factors that influence senior medical students to pursue careers in family medicine. Qualitative study using semistructured interviews. University of Western Ontario (UWO) in London. Eleven of 29 graduating UWO medical students matched to Canadian family medicine residency programs beginning in July 2001. Eleven semistructured interviews were conducted with a maximum variation sample of medical students. Interviews were transcribed and reviewed independently, and a constant comparative approach was used by the team to analyze the data. Family physician mentors were an important influence on participants' decisions to pursue careers in family medicine. Participants followed one of three pathways to selecting family medicine: from an early decision to pursue family medicine, from initial uncertainty about career choice, or from an early decision to specialize and a change of mind. The perception of a wide scope of practice attracts candidates to family medicine. Having more family medicine role models early in medical school might encourage more medical students to select careers in family medicine.


Two New Cynodonts (Therapsida) from the Middle-Early Late Triassic of Brazil and Comments on South American Probainognathians.
We describe two new cynodonts from the early Late Triassic of southern Brazil. One taxon, Bonacynodon schultzi gen. et sp. nov., comes from the lower Carnian Dinodontosaurus AZ, being correlated with the faunal association at the upper half of the lower member of the Chañares Formation (Ischigualasto-Villa Unión Basin, Argentina). Phylogenetically, Bonacynodon is a closer relative to Probainognathus jenseni than to any other probainognathian, bearing conspicuous canines with a denticulate distal margin. The other new taxon is Santacruzgnathus abdalai gen. et sp. nov. from the Carnian Santacruzodon AZ. Although based exclusively on a partial lower jaw, it represents a probainognathian close to Prozostrodon from the Hyperodapedon AZ and to Brasilodon, Brasilitherium and Botucaraitherium from the Riograndia AZ. The two new cynodonts and the phylogenetic hypothesis presented herein indicate the degree to which our knowledge on probainognathian cynodonts is incomplete and also the relevance of the South American fossil record for understanding their evolutionary significance. The taxonomic diversity and abundance of probainognathians from Brazil and Argentina will form the basis of deep and complex studies to address the evolutionary transformations of cynodonts leading to mammals.


Dental calculus reveals unique insights into food items, cooking and plant processing in prehistoric central Sudan.
Accessing information on plant consumption before the adoption of agriculture is challenging. However, there is growing evidence for use of locally available wild plants from an increasing number of pre-agrarian sites, suggesting broad ecological knowledge. The extraction of chemical compounds and microfossils from dental calculus removed from ancient teeth offers an entirely new perspective on dietary reconstruction, as it provides empirical results on material that is already in the mouth. Here we present a suite of results from the multi-period Central Sudanese site of Al Khiday. We demonstrate the ingestion in both pre-agricultural and agricultural periods of Cyperus rotundus tubers. This plant is a good source of carbohydrates and has many useful medicinal and aromatic qualities, though today it is considered to be the world's most costly weed. Its ability to inhibit Streptococcus mutans may have contributed to the unexpectedly low level of caries found in the agricultural population. Other evidence extracted from the dental calculus includes smoke inhalation, dry (roasting) and wet (heating in water) cooking, a second plant possibly from the Triticaceae tribe and plant fibres suggestive of raw material preparation through chewing.


Water contact angle is not a good predictor of biological responses to materials.
Often the view is expressed that water contact angle (WCA) or other wettability/surface energy measurements made on a material surface can be used to predict cellular attachment to materials, e.g., bacteria attach to hydrophobic surfaces. In this article, the authors present a perspective emerging from their work that has failed to find relationships between WCA and microbial and stem cell attachment within large diversity material libraries and compare with the literature concluding that such simple rules are (unfortunately) wholly inadequate to explain cell-material interactions.


Mollusks (Gastropoda and Bivalvia) of the Multiple-Use Reserve Martín García Island, Río de la Plata River: biodiversity and ecology.
The Island of Martin Garcia is located in the Upper Río de la Plata, to the south of mouth the Uruguay River. The aim of the present study was to analyse the biodiversity of the island freshwater mollusks and their relationships to environmental variables. Twelve sampling sites were selected, five were along the littoral section of the island and seven were Inland ponds. Seven major environmental variables were measured: water and air temperature, percentage of oxygen saturation, dissolved oxygen, electrical conductivity, total dissolved solids and pH. Twenty-seven mollusk species were found, Antillorbis nordestensis, Biomphalaria tenagophila tenagophila , B. t. guaibensis, B. straminea, B. peregrina, Drepanotrema kermatoides, D. cimex, D. depressissimum, Chilina fluminea, C. rushii, C. megastoma, Uncancylus concentricus, Hebetancylus moricandi, Stenophysa marmorata, Heleobia piscium, H. parchappii, Potamolithus agapetus, P. buschii, P. lapidum, Pomacea canaliculata, P. megastoma, Asolene platae, Corbicula fluminea, Eupera platensis, Pisidium sterkianum, P. taraguyense and Limnoperna fortunei. UPGMA clustering of species based on their occurrence in different ecological conditions revealed two main species groups. The Canonical Correspondence Analysis suggests that the species distribution is related to the physico-chemical condition of water. Axis two of the ordination diagram displayed the approximately 95.6% of the correlation between species and environmental variables. Dissolved oxygen, conductivity, water temperature and pH showed the highest fluctuations during the sampling period. The species richness (S) showed relationships mainly with water temperature and conductivity. The biodiversity of the gastropods and bivalves from Martín García Island amounts to up to 26 species. Among the Gastropoda, the Planorbidae family made the most sizeable contribution. The Lithogliphidae P. agapetus (26.28%) and P. buschii (9.50%) showed the highest relative frequencies of occurrence within the littoral environments, while the Planorbidae D. cimex (23.83%) and D. kermatoides (11.59%) likewise did so in the inland ponds.


Connexin 43 Controls the Astrocyte Immunoregulatory Phenotype.
Astrocytes are the most abundant glial cells of the central nervous system and have recently been recognized as crucial in the regulation of brain immunity. In most neuropathological conditions, astrocytes are prone to a radical phenotypical change called reactivity, which plays a key role in astrocyte contribution to neuroinflammation. However, how astrocytes regulate brain immunity in healthy conditions is an understudied question. One of the astroglial molecule involved in these regulations might be Connexin 43 (Cx43), a gap junction protein highly enriched in astrocyte perivascular endfeet-terminated processes forming the glia limitans. Indeed, Cx43 deletion in astrocytes (Cx43KO) promotes a continuous immune recruitment and an autoimmune response against an astrocyte protein, without inducing any brain lesion. To investigate the molecular basis of this unique immune response, we characterized the polysomal transcriptome of hippocampal astrocytes deleted for Cx43. Our results demonstrate that, in the absence of Cx43, astrocytes adopt an atypical reactive status with no change in most canonical astrogliosis markers, but with an upregulation of molecules promoting immune recruitment, complement activation as well as anti-inflammatory processes. Intriguingly, while several of these upregulated transcriptional events suggested an activation of the γ-interferon pathway, no increase in this cytokine or activation of related signaling pathways were found in Cx43KO. Finally, deletion of astroglial Cx43 was associated with the upregulation of several angiogenic factors, consistent with an increase in microvascular density in Cx43KO brains. Collectively, these results strongly suggest that Cx43 controls immunoregulatory and angiogenic properties of astrocytes.


Computation of arbitrarily constrained synthetic discriminant functions.
An algorithm for computing correlation filters based on synthetic discriminant functions that can be displayed on current spatial light modulators is presented. The procedure is nondivergent, computationally feasible, and capable of producing multiple solutions, thus overcoming some of the pitfalls of previous methods.


Effect of citicoline on functional and cognitive status among patients with traumatic brain injury: Citicoline Brain Injury Treatment Trial (COBRIT).
Traumatic brain injury (TBI) is a serious public health problem in the United States, yet no treatment is currently available to improve outcome after TBI. Approved for use in TBI in 59 countries, citicoline is an endogenous substance offering potential neuroprotective properties as well as facilitated neurorepair post injury. To determine the ability of citicoline to positively affect functional and cognitive status in persons with complicated mild, moderate, and severe TBI. The Citicoline Brain Injury Treatment Trial (COBRIT), a phase 3, double-blind randomized clinical trial conducted between July 20, 2007, and February 4, 2011, among 1213 patients at 8 US level 1 trauma centers to investigate effects of citicoline vs placebo in patients with TBI classified as complicated mild, moderate, or severe. Ninety-day regimen of daily enteral or oral citicoline (2000 mg) or placebo. Functional and cognitive status, assessed at 90 days using the TBI-Clinical Trials Network Core Battery. A global statistical test was used to analyze the 9 scales of the core battery. Secondary outcomes were functional and cognitive improvement, assessed at 30, 90, and 180 days, and examination of the long-term maintenance of treatment effects. Rates of favorable improvement for the Glasgow Outcome Scale-Extended were 35.4% in the citicoline group and 35.6% in the placebo group. For all other scales the rate of improvement ranged from 37.3% to 86.5% in the citicoline group and from 42.7% to 84.0% in the placebo group. The citicoline and placebo groups did not differ significantly at the 90-day evaluation (global odds ratio [OR], 0.98 [95% CI, 0.83-1.15]); in addition, there was no significant treatment effect in the 2 severity subgroups (global OR, 1.14 [95% CI, 0.88-1.49] and 0.89 [95% CI, 0.72-1.49] for moderate/severe and complicated mild TBI, respectively). At the 180-day evaluation, the citicoline and placebo groups did not differ significantly with respect to the primary outcome (global OR, 0.87 [95% CI, 0.72-1.04]). Among patients with traumatic brain injury, the use of citicoline compared with placebo for 90 days did not result in improvement in functional and cognitive status. clinicaltrials.gov Identifier: NCT00545662.


Near-IR fluorescence and reflectance confocal microscopy for imaging of quantum dots in mammalian skin.
Understanding the skin penetration of nanoparticles (NPs) is an important concern due to the increasing presence of NPs in consumer products, including cosmetics. Technical challenges have slowed progress in evaluating skin barrier and NP factors that contribute to skin penetration risk. To limit sampling error and other problems associated with histological processing, many researchers are implementing whole tissue confocal or multiphoton microscopies. This work introduces a fluorescence and reflectance confocal microscopy system that utilizes near-IR excitation and emission to detect near-IR lead sulfide quantum dots (QDs) through ex vivo human epidermis. We provide a detailed prediction and experimental analysis of QD detection sensitivity and demonstrate detection of QD skin penetration in a barrier disrupted model. The unique properties of near-IR lead-based QDs will enable future studies that examine the impact of further barrier-disrupting agents on skin penetration of QDs and elucidate mechanistic insight into QD tissue interactions at the cellular level.


High-resolution X-ray microtomography for the detection of lung tumors in living mice.
In the present study, the feasibility of applying high-resolution microtomography (micro-CT) for the detection of lung tumors was investigated in live mice at an early and more advanced stage of tumor development. The chest area of anesthesized mice was scanned by X-ray micro-CT. In mice with a minor and heavy tumor load, micro-CT proved to be a fast and noninvasive imaging device for the detection of lung tumors. After validation of the CT data by histologic sectioning, it was shown that the majority of tumors could be distinguished in the reconstructed virtual slices obtained by micro-CT. The data from micro-CT were also confirmed by visual inspection of the inflated and excised lungs postmortem. In vivo micro-CT opens broad perspectives for imaging tumor development and its progression in a noninvasive way. Micro-CT also allows for longitudinal evaluation of the treatment of lung cancer by drugs.


Trends in early initiation of antiretroviral therapy and characteristics of persons with HIV initiating therapy in San Francisco, 2007-2011.
In 2010, the San Francisco Department of Public Health offered antiretroviral therapy (ART) to all its patients with human immunodeficiency virus (HIV) regardless of CD4 count. We assessed trends in time from diagnosis to ART initiation and factors associated with ART initiation among San Francisco residents living with HIV between 2007 and 2011. Time to ART initiation decreased among those diagnosed with higher CD4 count. ART initiation rate was significantly higher in recent years and lower among African Americans, men who have sex with men who also inject drugs, and persons aged ≥50 years. We found a trend toward early treatment. However, racial and social disparities persist.


The relationship between vitamin A and risk of fracture: meta-analysis of prospective studies.
Osteoporotic fracture is a significant cause of morbidity and mortality and is a challenging global health problem. Previous reports of the relation between vitamin A intake or blood retinol and risk of fracture were inconsistent. We searched Medline and Embase to assess the effects of vitamin A (or retinol or beta-carotene but not vitamin A metabolites) on risk of hip and total fracture. Only prospective studies were included. We pooled data with a random effects meta-analysis with adjusted relative risk (adj.RR) and 95% confidence interval (CI). We used Q statistic and I(2) statistic to assess heterogeneity and Egger's test to assess publication bias. Eight vitamin A (or retinol or beta-carotene) intake studies (283,930 participants) and four blood retinol level prospective studies (8725 participants) were included. High intake of vitamin A and retinol were shown to increase risk of hip fracture (adj.RR [95% CI] = 1.29 [1.07, 1.57] and 1.40 [1.03, 1.91], respectively), whereas beta-carotene intake was not found to increase the risk of hip fracture (adj.RR [95% CI] = 0.82 [0.59, 1.14]). Both high or low level of blood retinol was shown to increase the risk of hip fracture (adj.RR [95% CI] = 1.87 [1.31, 2.65] and 1.56 [1.09, 2.22], respectively). The risk of total fracture does not differ significantly by level of vitamin A (or retinol) intake or by blood retinol level. Dose-response meta-analysis shows a U-shaped relationship between serum retinol level and hip fracture risk. Our meta-analysis suggests that blood retinol level is a double-edged sword for risk of hip fracture. To avoid the risk of hip fracture caused by too low or too high a level of retinol concentration, we suggest that intake of beta-carotene (a provitamin A), which should be converted to retinol in blood, may be better than intake of retinol from meat, which is directly absorbed into blood after intake.


Folic acid supplement use is the most significant predictor of folate concentrations in Canadian women of childbearing age.
One-fifth of Canadian women of childbearing age (WCBA) have red blood cell (RBC) folate concentrations below those considered optimal for neural tube defect risk reduction (≥906 nmol·L(-1)). Determinants of optimal concentrations have not been examined in a nationally representative sample of Canadian WCBA since food fortification with folic acid was implemented. This study explored correlates of optimal RBC folate concentrations and characteristics of folic acid supplement users in a sample of Canadian WCBA. RBC folate concentrations from the 2007-2009 Canadian Health Measures Survey were assessed in women aged 15 to 45 years (n = 1162). Sociodemographic, behavioural, and clinical determinants of RBC folate ≥906 nmol·L(-1) were examined using univariate and separate multiple logistic regression models that controlled for age and household income. t tests were used to study differences between folic acid supplement users and nonusers. WCBA not taking folic acid supplements were less likely to achieve a RBC folate concentration ≥906 nmol·L(-1) compared with folic acid supplement users (odds ratio, 0.47; 95% confidence interval, 0.24, 0.92). Twenty-five percent of WCBA reported folic acid supplement use, and there was a higher percentage of folic acid supplement users in the highest income group. Folic acid supplement users were also more frequent consumers of supplemental vitamin B(12) and of fruit and vegetables (>3 times per day). Folic acid supplement use was the most significant predictor of WCBA achieving optimal RBC folate concentrations. These results indicate a need for targeted strategies to improve compliance with folic acid supplement recommendations among WCBA.


Breakage-fusion-bridge cycles and large insertions contribute to the rapid evolution of accessory chromosomes in a fungal pathogen.
Chromosomal rearrangements are a major driver of eukaryotic genome evolution, affecting speciation, pathogenicity and cancer progression. Changes in chromosome structure are often initiated by mis-repair of double-strand breaks in the DNA. Mis-repair is particularly likely when telomeres are lost or when dispersed repeats misalign during crossing-over. Fungi carry highly polymorphic chromosomal complements showing substantial variation in chromosome length and number. The mechanisms driving chromosome polymorphism in fungi are poorly understood. We aimed to identify mechanisms of chromosomal rearrangements in the fungal wheat pathogen Zymoseptoria tritici. We combined population genomic resequencing and chromosomal segment PCR assays with electrophoretic karyotyping and resequencing of parents and offspring from experimental crosses to show that this pathogen harbors a highly diverse complement of accessory chromosomes that exhibits strong global geographic differentiation in numbers and lengths of chromosomes. Homologous chromosomes carried highly differentiated gene contents due to numerous insertions and deletions. The largest accessory chromosome recently doubled in length through insertions totaling 380 kb. Based on comparative genomics, we identified the precise breakpoint locations of these insertions. Nondisjunction during meiosis led to chromosome losses in progeny of three different crosses. We showed that a new accessory chromosome emerged in two viable offspring through a fusion between sister chromatids. Such chromosome fusion is likely to initiate a breakage-fusion-bridge (BFB) cycle that can rapidly degenerate chromosomal structure. We suggest that the accessory chromosomes of Z. tritici originated mainly from ancient core chromosomes through a degeneration process that included BFB cycles, nondisjunction and mutational decay of duplicated sequences. The rapidly evolving accessory chromosome complement may serve as a cradle for adaptive evolution in this and other fungal pathogens.


Structural and electronic properties of Co-corrole, Co-corrin, and Co-porphyrin.
A quantitative study of the structure and electronic properties of Co-corrole, Co-corrin, and Co-porphyrin, using density functional theory, is reported. The structure of each macrocycle is optimized, with no symmetry constraints, by considering different spin states. The ground-state structures and spin states (S = 1 for Co-corrole, S = 0 for Co-corrin and S = 1/2 for Co-porphyrin) are in good agreement with the experimental data available. The trends in the sizes of the coordination cavities upon varying the inner metal atom and/or the macrocycle are analyzed and compared with those for the Fe-porphyrin we studied previously. Our results reveal that most of the distortion of the Co-corrin core in the B12 coenzyme is due to the inherent properties of Co-corrin. Quite different behaviors are found between corrinoids and porphyrins upon varying the spin state. While an increase in the metal-nitrogen (M-N) distance with spin state occurs in the porphyrins, the corrinoids show little variation in the M-N distance and, in some cases, undergo small structural changes in the ring structure. These results aid in understanding the often discussed question of why nature has chosen corrin/porphyrin for carrying out the particular biological functions identified in the B12 coenzyme.


Asthma prevalence and risk factor assessment of an underserved and primarily Latino child population in Colorado.
Asthma is a substantialpublic health burden among children. Disease and risk-factor discrepancies have been identified among racial, ethnic, and socioeconomic groups. At a rural health clinic (Salud Family Health Center) with primarily underserved and Latino patients in Colorado, the authors evaluated 250 medical records and administered 57 parental surveys to describe this population with respect to asthma diagnosis, asthma-like symptoms, and environmental/occupational risk factors among children. Wheeze and asthma were indicated in 9.7% and 8.9% of medical records, respectively. Twenty parents (35.7%) reported in a questionnaire that their child had experienced wheezing or whistling in the chest. Parents reported that children play in farming fields (21.8%) and feed livestock/animals (10.9%). Additionally, 13.2% and 9.4% of children have a household member who works around livestock or around grain, feed, or dust, respectively. Information from the Salud population can be used to develop larger-scale research and public health initiatives to eliminate health and risk factor disparities among underserved children.


Magnetically-enabled biomarker extraction and delivery system: towards integrated ASSURED diagnostic tools.
Diagnosis of asymptomatic malaria poses a great challenge to global disease elimination efforts. Healthcare infrastructure in rural settings cannot support existing state-of-the-art tools necessary to diagnose asymptomatic malaria infections. Instead, lateral flow immunoassays (LFAs) are widely used as a diagnostic tool in malaria endemic areas. While LFAs are simple and easy to use, they are unable to detect low levels of parasite infection. We have developed a field deployable Magnetically-enabled Biomarker Extraction And Delivery System (mBEADS) that significantly improves limits of detection for several commercially available LFAs. Integration of mBEADS with leading commercial Plasmodium falciparum malaria LFAs improves detection limits to encompass an estimated 95% of the disease reservoir. This user-centered mBEADS platform makes significant improvements to a previously cumbersome malaria biomarker enrichment strategy by improving reagent stability, decreasing the processing time 10-fold, and reducing the assay cost 10-fold. The resulting mBEADS process adds just three minutes and less than $0.25 to the total cost of a single LFA, thus balancing sensitivity and practicality to align with the World Health Organization's ASSURED criteria for point-of-care (POC) testing.


Assessment of tobacco control advocacy behavioural capacity among students at schools of public health in China.
To evaluate student tobacco control advocacy behavioural capacity using longitudinal trace data. A tobacco control advocacy curriculum was developed and implemented at schools of public health (SPH) or departments of public health in seven universities in China. Participants comprised undergraduate students studying the public health curriculum in these 13 Universities. A standardised assessment tool was used to evaluate their tobacco control advocacy behavioural capacity. Repeated measures analysis of variance, paired t tests and paired χ² tests were used to determine differences between dependent variables across time. Multivariate analysis of variance (MANOVA) and multivariate logistic regression were used to assess treatment effects between intervention and control sites. Respective totals of 426 students in the intervention group and 338 in the control group were available for the evaluation. Approximately 90% of respondents were aged 21 years or older and 56% were women. Findings show that the capacity building program significantly improved public health student advocacy behavioural capacity, including advocacy attitude, interest, motivation and anti-secondhand smoke behaviours. The curriculum did not impact student smoking behaviour. This study provides sufficient evidence to support the implementation of tobacco control advocacy training at Chinese schools of public health.


Addition of human recombinant bone morphogenetic protein-2 to inactive commercial human demineralized freeze-dried bone allograft makes an effective composite bone inductive implant material.
COMMERCIAL PREPARATIONS OF HUMAN DEMINERALIZED freeze-dried bone allograft (DFDBA) vary in their ability to induce new bone formation. This study tested the hypothesis that inactive DFDBA can be used as an effective carrier of recombinant human bone morphogenetic protein-2 (rhBMP-2). Two batches of active DFDBA were used as controls. Two batches of DFDBA, previously shown to be inactive, were treated with vehicle or with 5 or 20 microg rhBMP-2 and implanted into the calf muscle of male Nu/Nu (nude) mice. Each mouse received one implant in each hind limb, both of which were of the same formulation, resulting in 8 groups of 4 mice per group: active DFDBA batch A, active DFDBA batch B, inactive DFDBA batch A, inactive DFDBA batch B, inactive DFDBA batch A plus 5 microg rhBMP-2, inactive DFDBA batch A plus 20 microg rhBMP-2, inactive DFDBA batch B plus 5 microg rhBMP-2, and inactive DFDBA batch B plus 20 microg rhBMP-2. After 56 days, the implants were removed and histologically examined. A semiquantitative bone induction index was calculated based on the amount of new bone covering each histological section. Histomorphometry was also used to evaluate the area of new bone formed and the area of residual implant material. The results showed that active DFDBA induces new bone formation, whereas inactive DFDBA does not. Addition of rhBMP-2 to inactive DFDBA results in new bone formation with a bone induction index comparable to that of active DFDBA. Histomorphometric analysis, however, revealed that the rhBMP-2-containing implants caused a dose-dependent increase in new bone area that exceeded that induced by active DFDBA. At the highest concentration of rhBMP-2, bone formation was exuberant. rhBMP-2 also caused the resorption of residual implant material to levels comparable to that seen in sites treated with active DFDBA, suggesting that this growth factor may regulate resorptive cells either directly or indirectly. This study shows that addition of rhBMP-2 to inactive DFDBA provides reproducible, consistent bone induction, and suggests that inactive commercial preparations may contain inadequate amounts of BMP to cause bone induction compared to active preparations.


Detecting the onset of accelerated long-term forgetting: evidence from temporal lobe epilepsy.
Accelerated long-term forgetting (ALF) refers to a slowly developing anterograde amnesia in which material is retained normally over short delays but then forgotten at an abnormally fast rate over days to weeks. Such long-term memory impairment is not detected by standard clinical tests. This study analysed ALF in a temporal lobe epileptic, RY. Key issues addressed were: (i) the timeframe of ALF onset; (ii) whether disruption of memory consolidation during sleep is a necessary requirement for precipitating ALF; (iii) the effectiveness of repeated recall in limiting the impact of ALF. RY's memory for novel word-pairings was compared with that of matched controls using cued-recall and forced choice recognition (FCR) tests at multiple delays (5, 30, 55, 240 min). To investigate the impact of repeated recall some pairings were recalled at all intervals, and all material (repeatedly and non-repeatedly recalled) was tested again after a 24h delay. RY's initial learning and performance at 30 min were normal, but by 55 min both his cued-recall performance and the subjective quality of his recognition memory were significantly impaired. This suggests disruption of secondary consolidation processes occurring relatively soon after learning. It also raises the possibility of developing a standard test to diagnose ALF within a single clinical session rather than requiring multiple visits. Since RY remained awake it appears that disruption of memory consolidation during sleep is not a necessary condition for him to experience ALF. Repeated recall at multiple time-points within the first 4h sustained normal recall performance to 24h, indicating repeated recall could form the basis for a protective strategy.


Nrf2 expression is regulated by epigenetic mechanisms in prostate cancer of TRAMP mice.
Nuclear factor-erythroid 2 p45-related factor 2 (Nrf2) is a transcription factor which regulates the expression of many cytoprotective genes. In the present study, we found that the expression of Nrf2 was suppressed in prostate tumor of the Transgenic Adenocarcinoma of Mouse Prostate (TRAMP) mice. Similarly, the expression of Nrf2 and the induction of NQO1 were also substantially suppressed in tumorigenic TRAMP C1 cells but not in non-tumorigenic TRAMP C3 cells. Examination of the promoter region of the mouse Nrf2 gene identified a CpG island, which was methylated at specific CpG sites in prostate TRAMP tumor and in TRAMP C1 cells but not in normal prostate or TRAMP C3 cells, as shown by bisulfite genomic sequencing. Reporter assays indicated that methylation of these CpG sites dramatically inhibited the transcriptional activity of the Nrf2 promoter. Chromatin immunopreceipitation (ChIP) assays revealed increased binding of the methyl-CpG-binding protein 2 (MBD2) and trimethyl-histone H3 (Lys9) proteins to these CpG sites in the TRAMP C1 cells as compared to TRAMP C3 cells. In contrast, the binding of RNA Pol II and acetylated histone H3 to the Nrf2 promoter was decreased. Furthermore, treatment of TRAMP C1 cells with DNA methyltransferase (DNMT) inhibitor 5-aza-2'-deoxycytidine (5-aza) and histone deacetylase (HDAC) inhibitor trichostatin A (TSA) restored the expression of Nrf2 as well as the induction of NQO1 in TRAMP C1 cells. Taken together, these results indicate that the expression of Nrf2 is suppressed epigenetically by promoter methylation associated with MBD2 and histone modifications in the prostate tumor of TRAMP mice. Our present findings reveal a novel mechanism by which Nrf2 expression is suppressed in TRAMP prostate tumor, shed new light on the role of Nrf2 in carcinogenesis and provide potential new directions for the detection and prevention of prostate cancer.


Hyperactivation of MAPK Signaling Is Deleterious to RAS/RAF-mutant Melanoma.
The most frequent genetic alterations in melanoma are gain-of-function (GOF) mutations in BRAF, which result in RAF-MEK-ERK signaling pathway addiction. Despite therapeutic success of RAF and MEK inhibitors in treating BRAFV600-mutant tumors, a major challenge is the inevitable emergence of drug resistance, which often involves reactivation of the MAPK pathway. Interestingly, resistant tumors are often sensitive to drug withdrawal, suggesting that hyperactivation of the MAPK pathway is not tolerated. To further characterize this phenomenon, isogenic models of inducible MAPK hyperactivation in BRAFV600E melanoma cells were generated by overexpression of ERK2. Using this model system, supraphysiologic levels of MAPK signaling led to cell death, which was reversed by MAPK inhibition. Furthermore, complete tumor regression was observed in an ERK2-overexpressing xenograft model. To identify mediators of MAPK hyperactivation-induced cell death, a large-scale pooled shRNA screen was conducted, which revealed that only shRNAs against BRAF and MAP2K1 rescued loss of cell viability. This suggested that no single downstream ERK2 effector was required, consistent with pleiotropic effects on multiple cellular stress pathways. Intriguingly, the detrimental effect of MAPK hyperactivation could be partially attributed to secreted factors, and more than 100 differentially secreted proteins were identified. The effect of ERK2 overexpression was highly context dependent, as RAS/RAF mutant but not RAS/RAF wild-type melanoma were sensitive to this perturbation. IMPLICATIONS: This vulnerability to MAPK hyperactivation raises the possibility of novel therapeutic approaches for RAS/RAF-mutant cancers.


Simulation of anisotropic growth of low-grade gliomas using diffusion tensor imaging.
A recent computational model of brain tumor growth, developed to better describe how gliomas invade through the adjacent brain parenchyma, is based on two major elements: cell proliferation and isotropic cell diffusion. On the basis of this model, glioma growth has been simulated in a virtual brain, provided by a 3D segmented MRI atlas. However, it is commonly accepted that glial cells preferentially migrate along the direction of fiber tracts. Therefore, in this paper, the model has been improved by including anisotropic extension of gliomas. The method is based on a cell diffusion tensor derived from water diffusion tensor (as given by MRI diffusion tensor imaging). Results of simulations have been compared with two clinical examples demonstrating typical growth patterns of low-grade gliomas centered around the insula. The shape and the kinetic evolution are better simulated with anisotropic rather than isotropic diffusion. The best fit is obtained when the anisotropy of the cell diffusion tensor is increased to greater anisotropy than the observed water diffusion tensor. The shape of the tumor is also influenced by the initial location of the tumor. Anisotropic brain tumor growth simulations provide a means to determine the initial location of a low-grade glioma as well as its cell diffusion tensor, both of which might reflect the biological characteristics of invasion.


Internet and written respiratory questionnaires yield equivalent results for adolescents.
This study compared results from Internet and written questionnaires about respiratory symptoms in order to find out if both forms of the survey yielded the same answers. One thousand seventy-one students, ages 13 to 17, were asked to complete either an Internet or a written questionnaire. The demographic characteristics of the participants equalled those of the general Dutch adolescent population. Participants were randomly assigned to fill out an electronic or written questionnaire. In addition to eight items from the International Study of Asthma and Allergies in Childhood (ISAAC) questionnaire, two items on doctor visits (medical attention) regarding asthma or allergic disease during the past 12 months were included. The participation rate was 87%. The Internet version of the questionnaire showed fewer missing answers than the written version, but this was not statistically significant. The respiratory items did not show statistically significant score differences between the Internet and written modes of administration, and there was no visible trend for higher respectively lower scores by either mode of questionnaire administration. From these results, we conclude that respiratory questionnaires may be provided to adolescents electronically rather than on paper, since both approaches yielded equal results. To generalize these findings, we recommend repeated studies in other settings.


Value of early referral to fertility preservation in young women with breast cancer.
To determine whether early referral to reproductive specialists improves fertility preservation (FP) outcomes and reduces delay in adjuvant treatment in young women with breast cancer. A secondary analysis of a prospective database of patients with breast cancer undergoing ovarian stimulation (OS) for FP by oocyte or embryo cryopreservation was performed. Of the 154 patients, 93 met the inclusion criteria (mean age, 35.2 ± 4.4 years). Thirty-five of the 93 patients were referred before breast surgery (PreS), and 58 patients were referred after surgery (PostS). The time periods from initial diagnosis (ID) to initiation of OS (42.6 ± 27.7 days for PreS v 71.9 ± 30.7 days for PostS; P < .001) and from ID to initiation of chemotherapy (83.9 ± 24.3 days for PreS v 107.8 ± 42.9 days for PostS; P = .045) were significantly shorter for the PreS group versus the PostS group. Nine (25.7%) of 35 patients in the PreS group versus one (1.7%) of 58 patients in the PostS group were able to undergo two FP cycles (P < .001), resulting in an increased yield of oocytes in the PreS group (18.2% [93 of 511 oocytes] v 0.6% [five of 800 oocytes], respectively; P < .001) and embryos (17.2% [40 of 233 embryos] v 0.6% [two of 357 embryos], respectively; P < .001). Patients who had an oocyte retrieval within 5 weeks of the surgery were able to complete a second cycle within 9 weeks of the surgery. FP referral before breast surgery enables earlier initiation of cryopreservation cycles and chemotherapy and, when appropriate, multiple FP cycles. Women who can undergo multiple cycles may be at advantage for FP because of a larger number of oocytes or embryos cryopreserved. This is the first study demonstrating the benefit of early FP referral in patients with cancer.


Microbial granulation for lactic acid production.
This work investigated the formation of microbial granules to boost the productivity of lactic acid (LA). The flocculated form of LA-producing microbial consortium, dominated by Lactobacillus sp. (91.5% of total sequence), was initially obtained in a continuous stirred-tank reactor (CSTR), which was fed with 2% glucose and operated at a hydraulic retention time (HRT) of 12 h and pH 5.0 ± 0.1 under a thermophilic condition (50°C). The mixed liquor in the CSTR was then transferred to an up-flow anaerobic sludge blanket reactor (UASB). The fermentation performance and granulation process were monitored with a gradual decrease of HRT from 8.0 to 0.17 h, corresponding to an increase in the substrate loading from 60 to 2,880 g glucose L(-1) d(-1) . As the operation continued, the accumulation of biomass in the UASB was clearly observed, which changed from flocculent to granular form with decrease in HRT. Up to the HRT decrease to 0.5 h, the LA concentration was maintained at 19-20 g L(-1) with over 90% of substrate removal efficiency. However, further decrease of HRT resulted in a decrease of LA concentration with increase in residual glucose. Nevertheless, the volumetric LA productivity continuously increased, reaching 67 g L-fermenter (-1) h(-1) at HRT 0.17 h. The size of LA-producing granules and hydrophobicity gradually increased with decrease in HRT, reaching 6.0 mm and 60%, respectively. These biogranules were also found to have high settling velocities and low porosities, ranging 2.69-4.73 cm s(-1) and 0.39-0.92, respectively.


Patterns of Cardiometabolic Health as Midlife Women Transition to Menopause: A Prospective Multiethnic Study.
Cardiometabolic conditions increase in midlife, but early customized prevention strategies are not established for such women. To characterize and identify factors longitudinally related to constellations of cardiometabolic risk components in multiracial/ethnic women in midlife. We conducted a prospective, longitudinal, multiethnic cohort study of 3003 midlife women undergoing menopausal transition (MT). Metabolic syndrome (MetS) was defined as having at least three of five components: high fasting triglyceride (hTG) level, low high-density lipoprotein cholesterol (lHDL-C) level, high fasting plasma glucose (hGluc) level, large waist circumference (abdominal obesity; Ob), and hypertension (HTN). We described the patterns of constellations and estimated hazard ratios (HRs) for constellations at (i) incident MetS and (ii) recovery from MetS, using multivariable-adjusted Cox regression. Seven US sites. In all, 1412 non-Hispanic white, 851 black, 272 Japanese, 237 Hispanic, and 231 Chinese women. Race/ethnicity, lifestyle factors, and MT stage. Cardiometabolic constellations, incident MetS, and MetS recovery. Central obesity was the most frequent component. Having no components was the most frequent (31%) baseline constellation. Physical activity (HR = 1.68; 95% CI: 1.06, 2.68) and lower caloric intake (HR = 0.96; 95% CI: 0.93, 0.99 per 100 cal/d) were associated with recovery from MetS. Ob/hTG/lHDL-C (18%), Ob/HTN/lHDL-C (16%), and Ob/HTN/hGluc (14%) were frequent incident constellations. Physically active women had 26% to 62% lower hazards of incident MetS than inactive women. Modifiable lifestyle behaviors were related to recovery from MetS and decreased risk of the most frequent MetS constellations in midlife women.