Preliminary biometric analysis of mesiodistal tooth dimensions in subjects with normal occlusion.
The aims of this study were to confirm the efficacy of the Bolton index in a group of natural Class I white Italian patients with complete dentition, minimal crowding, and no previous orthodontic treatment, and to evaluate the mesiodistal tooth dimensions with multivariate cluster analysis. Tooth measurements were obtained from a sample of 56 patients with normal occlusion, with a normal Bolton index (22 men, 34 women; mean age, 27.8 years), without previous orthodontic treatment or prosthetic or Class II restorations. Maxillary and mandibular measurements were analyzed separately in the sexes. Clustering was performed using the partitioning around medoid algorithm and principal component analysis-based transformed data. Statistical analysis was conducted. The multivariate analysis showed 3 distinct clusters of both maxillary and mandibular tooth measurements in the male and female subjects. Statistically significant differences were found between the sexes in terms of average tooth measurements, and there was significant proportionality between the maxillary and mandibular arch clusters. Although the Bolton index is useful for identifying dentodental discrepancies in most patients, cluster analysis enabled the mandibular and maxillary tooth dimensions of the male and female subjects to be divided into 3 general classes (clusters) and the precise location of the discrepancies to be pinpointed.


A novel mechanism for the facilitation of theta-induced long-term potentiation by brain-derived neurotrophic factor.
Brain-derived neurotrophic factor (BDNF) contributes to the induction of long-term potentiation (LTP) by theta-pattern stimulation, but the specific processes underlying this effect are not known. Experiments described here, using BDNF concentrations that have minor effects on baseline responses, show that the neurotrophin both reduces the threshold for LTP induction and elevates the ceiling on maximal potentiation. The enhanced LTP proved to be as stable and resistant to reversal as that recorded under control conditions. BDNF markedly increased the facilitation of burst responses that occurs within a theta train. This suggests that the neurotrophin acts on long-lasting events that (1) are set in motion by the first burst in a train and (2) regulate the amplitude of subsequent bursts. Whole-cell recordings established that BDNF causes a rapid reduction in the size of the long-lasting afterhyperpolarization (AHP) that follows individual theta bursts. Apamin, an antagonist of type 2 small-conductance Ca2+-activated potassium (SK2) channels, also reduced hippocampal AHPs and closely reproduced the effects of BDNF on theta-burst responses and LTP. The latter results were replicated with a newly introduced, highly selective inhibitor of SK2 channels. Immunoblot analyses indicated that BDNF increases SK2 serine phosphorylation in hippocampal slices. These findings point to the conclusion that BDNF-driven protein kinase cascades serve to depress the SK2 component, and possibly other constituents, of the AHP. It is likely that this mechanism, acting with other factors, promotes the formation and increases the magnitude of LTP.


Structural analysis of the human tibia by tomographic (pQCT) serial scans.
This study analyses the evaluation of tomographic indicators of tibia structure, assuming that the usual loading pattern shifts from uniaxial compression close to the heel to a combined compression, torsion and bending scheme towards the knee. To this end, pQCT scans were obtained at 5% intervals of the tibia length (S5-S95 sites from heel to knee) in healthy men and women (10/10) aged 20-40 years. Indicators of bone mass [cortical area, cortical/total bone mineral content (BMC)], diaphyseal design (peri/endosteal perimeters, cortical thickness, circularity, bending/torsion moments of inertia - CSMIs), and material quality [(cortical vBMD (bone mineral density)] were determined. The longitudinal patterns of variation of these measures were similar between genders, but male values were always higher except for cortical vBMD. Expression of BMC data as percentages of the minimal values obtained along the bone eliminated those differences. The correlative variations in cortical area, BMC and thickness, periosteal perimeter and CSMIs along the bone showed that cortical bone mass was predominantly associated with cortical thickness toward the mid-diaphysis, and with bone diameter and CSMIs moving more proximally. Positive relationships between CSMIs (y) and total BMC (x) showed men's values shifting to the upper-right region of the graph and women's values shifting to the lower-left region. Total BMC decayed about 33% from S5 to S15 (where minimum total BMC and CSMI values and variances and maximum circularity were observed) and increased until S45, reaching the original S5 value at S40. The observed gender-related differences reflected the natural allometric relationships. However, the data also suggested that men distribute their available cortical mass more efficiently than women. The minimum amount and variance of mass indicators and CSMIs, and the largest circularity observed at S15 reflected the assumed adaptation to compression pattern at that level. The increase in CSMIs (successively for torsion, A-P bending, and lateral bending), the decrease in circularity values and the changes in cortical thickness and periosteal perimeter toward the knee described the progressive adaptation to increasing torsion and bending stresses. In agreement with the biomechanical background, the described relationships: (i) identify the sites at which some changes in tibial stresses and diaphyseal structure take place, possibly associated with fracture incidence; (ii) allow prediction of mass indicators at any site from single determinations; (iii) establish the proportionality between the total bone mass at regions with highly predominant trabecular and cortical bone of the same individual, suitable for a specific evaluation of changes in trabecular mass; and (iv) evaluate the ability of bone tissue to self-distribute the available cortical bone according to specific stress patterns, avoiding many anthropometric and gender-derived influences.


Association between Australian adolescent alcohol use and alcohol use risk and protective factors in 2011 and 2014.
Changes in risk and protective factors of adolescent alcohol use may be contributing to the recent decline in Australian adolescents alcohol use. The study aimed to determine the: (i) prevalence of alcohol use, risk and protective factors in 2011 and 2014; and (ii) association between alcohol use and risk and protective factors in 2011 and 2014. A repeat cross-sectional study was conducted. Grade 9-10 (aged 15-17 years) students from 32 Australian secondary schools were sampled in 2011 and 2014. A self-report survey collected data regarding alcohol use (ever, recent, 'binge drinking'), risk factors (e.g. alcohol use/permissive attitude to alcohol by friends/siblings/parents) and protective factors (e.g. self-efficacy; school/home/community support; peer caring relationships). Descriptive statistics were used to determine differences in alcohol use, risk and protective factors between 2011 and 2014. Adjusted multivariable logistic regression analyses examined associations between alcohol use, risk and protective factors separately in 2011 and 2014 (six models). Fewer adolescents reported alcohol use in 2014 compared with 2011 (ever: 56.6% vs. 67.9%, recent: 17.3% vs. 21.2%, 'binge drinking': 20.0% vs. 23.5%; 2011: n = 4366; 2014: n = 5199). Significant differences between 2014 and 2011 were found for some risk (five lower; one higher) and protective factors (four lower). Risk factors that were significantly lower in 2014 compared to 2011 were amongst variables with the strongest associations with alcohol use. The strength of associations with alcohol use, and decrease in the prevalence of certain risk factors in 2014 compared to 2011, suggests such factors may be contributing to the decline in adolescent alcohol use.


Clinical perspectives on secular trends of intervertebral foramen diameters in an industrialized European society.
Low back pain origins have been a matter of great controversy. While spinal stenosis is now radiologically traceable, the alteration of intervertebral foramen is less clear. The aim of this study was to assess "secular trends"-alterations occurring from one generation to the next-in osseous intervertebral foramina of the major vertebral segments in an industrialized society, and to discuss their possible clinical implication. The macerated "maximum intervertebral foramen width" and "intervertebral foramen height" of all major vertebral levels in 71 non-pathologic Swiss adult skeletons from the nineteenth and early twentieth century, with known individual age and sex and similar geographic and socio-economic background, were measured by sliding caliper at validated landmarks. A secular trend of the increase in "maximum intervertebral foramen width" is found for most levels, with females showing a more prominent alteration. Additionally, the non-pathologic "maximum intervertebral foramen width" does not change with respect to individual age, nor is a significant side difference detectable. "Intervertebral foramen height," hereby defined as the difference of the dorsal vertebral body height minus pedicle height, demonstrates for most levels, and either sex, an insignificant negative secular trend. Neither stature nor skeletal robustness vary significantly through time within this particular sample. The results of this study, despite obvious inadequacies of methods used, exclude secular narrowing of the "maximum intervertebral foramen width" as the only cause of radiculopathy or spinal stenosis. Furthermore, we found a mild insignificant decrease of the clinically more relevant "intervertebral foramen height." Nevertheless, the detected short-time variability of the bony intervertebral foramen, independent of individual stature, skeletal robustness or age, argues for an enhanced focus on the understanding of clinically relevant changes of spinal morphology from generation to generation.


Cow's milk consumption, HLA-DQB1 genotype, and type 1 diabetes: a nested case-control study of siblings of children with diabetes. Childhood diabetes in Finland study group.
The evidence for the putative role of cow's milk in the development of type 1 diabetes is controversial. We studied infant feeding patterns and childhood diet by structured questionnaire (n = 725) and HLA-DQB1 genotype by a polymerase chain reaction-based method (n = 556) in siblings of affected children and followed them for clinical type 1 diabetes. In a nested case-control design in a population who had both dietary and genetic data available, we selected as cases those siblings who progressed to clinical diabetes during the follow-up period (n = 33). For each case, we chose as matched control subjects siblings who fulfilled the following criteria: same sex, age within 1 year, not from the same family, the start of the follow-up within 6 months of that of the respective case, and being at risk for type 1 diabetes at the time the case presented with that disease (n = 254). The median follow-up time was 9.7 years (range 0.2-11.3). Early age at introduction of cow's milk supplements was not significantly associated with progression to clinical type 1 diabetes (relative risk adjusted for matching factors, maternal education, maternal and child's ages, childhood milk consumption, and genetic susceptibility markers was 1.60 [95% CI 0.5-5.1]). The estimated relative risk of childhood milk consumption for progression to type 1 diabetes was 5.37 (1.6-18.4) when adjusted for the matching and aforementioned sociodemographic factors, age at introduction of supplementary milk feeding, as well as for genetic susceptibility markers. In conclusion, our results provide support for the hypothesis that high consumption of cow's milk during childhood can be diabetogenic in siblings of children with type 1 diabetes. However, further studies are needed to assess the possible interaction between genetic disease susceptibility and dietary exposures in the development of this disease.


Unintended pregnancy and subsequent use of modern contraceptive among slum and non-slum women in Nairobi, Kenya.
In spite of major gains in contraceptive prevalence over the last few decades, many women in most parts of the developing world who would like to delay or avoid pregnancy do not use any method of contraception. This paper seeks to: a) examine whether experiencing an unintended pregnancy is associated with future use of contraception controlling for a number factors including poverty at the household and community levels; and b) investigate the mechanisms through which experiencing an unintended pregnancy leads to uptake of contraception. Quantitative and qualitative data from a cross-sectional research project conducted in 2009/10 in two slum settlements and two non-slum settings of Nairobi, Kenya are used. The quantitative component of the project was based on a random sample of 1,259 women aged 15-49 years. Logistic regression models were used to assess the effect of unintended pregnancy on future contraceptive use. The qualitative component of the project successfully interviewed a total of 80 women randomly selected from survey participants who had reported having at least one unintended pregnancy. Women whose last pregnancy was unintended were more likely to be using a modern method of contraception, compared to their peers whose last pregnancy was intended, especially among the wealthier group as shown in the interaction model. Among poor women, unintended pregnancy was not associated with subsequent use of contraception. The qualitative investigation with women who had an unplanned pregnancy reveals that experiencing an unintended pregnancy seems to have served as a "wake-up call", resulting in greater attention to personal risks, including increased interest in pregnancy prevention. For some women, unintended pregnancy was a consequence of strong opposition by their partners to family planning, while others reported they started using contraceptives following their unintended pregnancy, but discontinued after experiencing side effects. This study provides quantitative and qualitative evidence that women who have had an unintended pregnancy are "ready for change". Family planning programs may use the contacts with antenatal, delivery and post-delivery care system as an opportunity to identify women whose pregnancy is unplanned, and target them with information and services.


Activation of complement by serum-resistant Neisseria gonorrhoeae. Assembly of the membrane attack complex without subsequent cell death.
Interaction of the human complement system in normal human serum (NHS) with serum-resistant and -sensitive Neisseria gonorrhoeae was evaluated to better understand the mechanism of serum-resistance. Complement activity (CH50) was depleted from NHS in a dose-dependent fashion by both serum-resistant and -sensitive N. gonorrhoeae. No detectable CH50 remained in NHS incubated with 10(9) colony-forming units (CFU)/ml serum of either resistant or sensitive strains. When smaller numbers of bacteria were incubated with NHS, lesser, yet comparable, amounts of CH50 were depleted by both resistant and sensitive strains. Hemolytic C2 activity was diminished by 33% in the case of resistant N. gonorrhoeae (10(8) CFU/ml serum) and by 48% in the case of a sensitive strain. No detectable decreases in hemolytic C4 or C7 activities were found with either sensitive or resistant strains at this concentration. Both resistant and sensitive strains activated C1s in NHS. Resistant strains specifically activated 19-21% of radiolabeled C1s in NHS, whereas sensitive strains activated 18-32%. Both resistant and sensitive strains also activated C5 in NHS. In binding assays using radiolabeled C5 and C9 in NHS, resistant and sensitive strains bound comparable amounts of C5 and C9. The number of bound C5 and C9 molecules varied according to the number of bacteria or amount of serum used in the assay. The ratio of C9/C5 bound to a sensitive strain was 6.8, and to a resistant strain was 8.2, suggesting that C5 and C9 were incorporated into membrane attack complexes (MAC). Electron microscopic examination of resistant and sensitive strains incubated with NHS revealed that MAC is bound to the surfaces of the resistant strain as well as the sensitive strain.


An Innovative Framework to Improve Teratogenic Medication Risk Counseling.
Health care providers in the Veterans Affairs system are caring for a growing number of women of childbearing age. Prior research shows that a significant number of women veterans are prescribed category D or X teratogenic medications. However, the majority do not receive teratogenic risk or contraceptive counseling. In order to improve teratogenic medication prescribing practices at the Veterans Affairs Pittsburgh Healthcare System, we developed a framework called TARCC to guide primary care providers through appropriate teratogenic risk counseling. The components of TARCC are teratogen (Are you prescribing a potential teratogen?), alternative (Did you consider a nonteratogenic alternative?), risks (Did you discuss the risks of the medication?), contraception (Did you discuss contraception?), and chart (Did you chart your discussion?). We incorporated TARCC into an electronic health record reminder. Additional probes, educational tools, and options to place consults and orders are included within the reminder. The Teratogen Pharmacy E-consult prompts a pharmacist to electronically review a patient's medications and provide recommendations regarding potential alternatives and discussions of risks and benefits. This article describes the development and implementation of the TARCC framework in a clinical reminder.


The stepwise generation of multimetallic complexes based on a vinylbipyridine linkage and their photophysical properties.
The versatile rhenium complex [ReCl(CO)3(bpyC[triple bond, length as m-dash]CH)] (HC[triple bond, length as m-dash]Cbpy = 5-ethynyl-2,2'-bipyridine) is used to generate a series of bimetallic complexes through the hydrometallation of [MHCl(CO)(BTD)(PPh3)2] (M = Ru, Os; BTD = 2,1,3-benzothiadiazole). The ruthenium complex [Ru{CH[double bond, length as m-dash]CH-bpyReCl(CO)3}Cl(BTD)(CO)(PPh3)2] was characterised structurally. Ligand exchange reactions with bifunctional linkers bearing oxygen and sulfur donors provide access to tetra- and pentametallic complexes such as [{M{CH[double bond, length as m-dash]CH-bpyReCl(CO)3}(CO)(PPh3)2}2(S2CNC4H8NCS2)] and Fe[C5H4CO2M{CH[double bond, length as m-dash]CH-bpyReCl(CO)3}(CO)(PPh3)2]2. The effect of the group 8 metal on the photophysical properties of the rhenium centre was investigated using the complexes [Ru{CH[double bond, length as m-dash]CH-bpyReCl(CO)3}Cl(BTD)(CO)(PPh3)2] and [M{CH[double bond, length as m-dash]CH-bpyReCl(CO)3}{S2P(OEt)2}(CO)(PPh3)2] (M = Ru, Os). This revealed the quenching of the rhenium-based emission in favour of weak radiative processes based on the Ru and Os centres. The potential for exploiting this effect is illustrated by the reaction of [Ru{CH[double bond, length as m-dash]CH-bpyReCl(CO)3}Cl(CO)(BTD)(PPh3)2] with carbon monoxide, which results in a 5-fold fluorescence enhancement in the dicarbonyl product, [Ru{CH[double bond, length as m-dash]CH-bpyReCl(CO)3}Cl(CO)2(PPh3)2], as the quenching effect is disrupted.


The malaria testing and treatment landscape in Kenya: results from a nationally representative survey among the public and private sector in 2016.
Since 2004, Kenya's national malaria treatment guidelines have stipulated artemisinin-based combination therapy (ACT) as first-line treatment for uncomplicated malaria, and since 2014, confirmatory diagnosis of malaria in all cases before treatment has been recommended. A number of strategies to support national guidelines have been implemented in the public and private sectors in recent years. A nationally-representative malaria outlet survey, implemented across four epidemiological zones, was conducted between June and August 2016 to provide practical evidence to inform strategies and policies in Kenya towards achieving national malaria control goals. A total of 17,852 outlets were screened and 2271 outlets were eligible and interviewed. 78.3% of all screened public health facilities stocked both malaria diagnostic testing and quality-assured ACT (QAACT). Sulfadoxine-pyrimethamine (SP) for intermittent preventive treatment in pregnancy was available in 70% of public health facilities in endemic areas where it is recommended for treatment. SP was rarely found in the public sector outside of the endemic areas (< 0.5%). The anti-malaria stocking private sector had lower levels of QAACT (46.7%) and malaria blood testing (20.8%) availability but accounted for majority of anti-malarial distribution (70.6% of the national market share). More than 40% of anti-malarials were distributed by unregistered pharmacies (37.3%) and general retailers (7.1%). QAACT accounted for 58.2% of the total anti-malarial market share, while market share for non-QAACT was 15.8% and for SP, 24.8%. In endemic areas, 74.9% of anti-malarials distributed were QAACT. Elsewhere, QAACT market share was 49.4% in the endemic-prone areas, 33.2% in seasonal-transmission areas and 37.9% in low-risk areas. Although public sector availability of QAACT and malaria diagnosis is relatively high, there is a gap in availability of both testing and treatment that must be addressed. The private sector in Kenya, where the majority of anti-malarials are distributed, is also critical for achieving universal coverage with appropriate malaria case management. There is need for a renewed commitment and effective strategies to ensure access to affordable QAACT and confirmatory testing in the private sector, and should consider how to address malaria case management among informal providers responsible for a substantial proportion of the anti-malarial market share.


Multiple splicing defects caused by hERG splice site mutation 2592+1G>A associated with long QT syndrome.
Long QT syndrome type 2 (LQT2) is caused by mutations in the human ether-a-go-go-related gene (hERG). Cryptic splice site activation in hERG has recently been identified as a novel pathogenic mechanism of LQT2. In this report, we characterize a hERG splice site mutation, 2592+1G>A, which occurs at the 5' splice site of intron 10. Reverse transcription-PCR analyses using hERG minigenes transfected into human embryonic kidney-293 cells and HL-1 cardiomyocytes revealed that the 2592+1G>A mutation disrupted normal splicing and caused multiple splicing defects: the activation of cryptic splice sites within exon 10 and intron 10 and complete intron 10 retention. We performed functional and biochemical analyses of the major splice product, hERGΔ24, in which 24 amino acids within the cyclic nucleotide binding domain of the hERG channel COOH-terminus is deleted. Patch-clamp experiments revealed that the splice mutant did not generate hERG current. Western blot and immunostaining studies showed that mutant channels did not traffic to the cell surface. Coexpression of wild-type hERG and hERGΔ24 resulted in significant dominant-negative suppression of hERG current via the intracellular retention of the wild-type channels. Our results demonstrate that 2592+1G>A causes multiple splicing defects, consistent with the pathogenic mechanisms of long QT syndrome.


New insights into atopic dermatitis: role of skin barrier and immune dysregulation.
Atopic dermatitis (AD) is a chronic inflammatory skin disease that is often associated with the development of food allergy and asthma. New insights into AD reveals an important role for structural abnormalities in the epidermis resulting in a leaky epithelial barrier as well as chronic immune activation that contribute to the pathophysiology of this common skin disease. Patients with AD have a predisposition to colonization or infection by microbial organisms, most notably Staphylococcus aureus and herpes simplex virus (HSV). Measures directed at healing and protecting the skin barrier and controlling the immune activation are needed for effective management of AD. Early intervention may improve outcomes for AD as well as reduce the systemic allergen sensitization that may lead to associated allergic diseases in other organs.


Pollutant removal in a multi-stage municipal wastewater treatment system comprised of constructed wetlands and a maturation pond, in a temperate climate.
A multi-stage municipal wastewater treatment system is proposed to comply with Mexican standards for discharge into receiving water bodies. The system is located in Santa Fe de la Laguna, Mexico, an area with a temperate climate. It was designed for 2,700 people equivalent (259.2 m3/d) and consists of a preliminary treatment, a septic tank as well as two modules operating in parallel, each consisting of a horizontal subsurface-flow wetland, a maturation pond and a vertical flow polishing wetland. After two years of operation, on-site research was performed. An efficient biochemical oxygen demand (BOD5) (94-98%), chemical oxygen demand (91-93%), total suspended solids (93-97%), total Kjeldahl nitrogen (56-88%) and fecal coliform (4-5 logs) removal was obtained. Significant phosphorus removal was not accomplished in this study (25-52%). Evapotranspiration was measured in different treatment units. This study demonstrates that during the dry season wastewater treatment by this multi-stage system cannot comply with the limits established by Mexican standards for receiving water bodies type 'C'. However, it has demonstrated the system's potential for less restrictive uses such as agricultural irrigation, recreation and provides the opportunity for wastewater treatment in rural areas without electric energy.


Sodium valproate induces mitochondrial respiration dysfunction in HepG2 in vitro cell model.
Sodium valproate (VPA) is a potentially hepatotoxic antiepileptic drug. Risk of VPA-induced hepatotoxicity is increased in patients with mitochondrial diseases and especially in patients with POLG1 gene mutations. We used a HepG2 cell in vitro model to investigate the effect of VPA on mitochondrial activity. Cells were incubated in glucose medium and mitochondrial respiration-inducing medium supplemented with galactose and pyruvate. VPA treatments were carried out at concentrations of 0-2.0mM for 24-72 h. In both media, VPA caused decrease in oxygen consumption rates and mitochondrial membrane potential. VPA exposure led to depleted ATP levels in HepG2 cells incubated in galactose medium suggesting dysfunction in mitochondrial ATP production. In addition, VPA exposure for 72 h increased levels of mitochondrial reactive oxygen species (ROS), but adversely decreased protein levels of mitochondrial superoxide dismutase SOD2, suggesting oxidative stress caused by impaired elimination of mitochondrial ROS and a novel pathomechanism related to VPA toxicity. Increased cell death and decrease in cell number was detected under both metabolic conditions. However, immunoblotting did not show any changes in the protein levels of the catalytic subunit A of mitochondrial DNA polymerase γ, the mitochondrial respiratory chain complexes I, II and IV, ATP synthase, E3 subunit dihydrolipoyl dehydrogenase of pyruvate dehydrogenase, 2-oxoglutarate dehydrogenase and glutathione peroxidase. Our results show that VPA inhibits mitochondrial respiration and leads to mitochondrial dysfunction, oxidative stress and increased cell death, thus suggesting an essential role of mitochondria in VPA-induced hepatotoxicity.


Validation of the 2-DeltaDeltaCt calculation as an alternate method of data analysis for quantitative PCR of BCR-ABL P210 transcripts.
Chronic myelogenous leukemia (CML) is a clonal myeloproliferative disorder that is characterized by the presence of a reciprocal translocation between chromosomes 9 and 22 and results in the formation of the Philadelphia (Ph1) chromosome and is present in most of CML patients. The Ph1 chromosome forms a chimeric gene that encodes an abnormal P210 mRNA transcript in most CML patients. Surveillance for minimal residual disease by detection of BCR/ABL transcripts is currently done mostly by quantitative real-time reverse transcriptase polymerase chain reaction (RT-PCR). Quantitation of BCR/ABL transcripts can monitor tumor load and the outcome of therapy. Absolute quantification determines the input copy number of the transcript of interest, usually by plotting the amount of PCR product onto a standard curve based on serial dilutions of the same product cloned in plasmids. Relative quantification describes the change in expression of the target gene in the patient sample relative to that of a control transcript by using the 2-DeltaDeltaCt calculation. The results of real-time RT-PCR for BCR/ABL transcripts are often analyzed by using plasmid DNA standard curves. In the present study, 79 BCR/ABL transcript-positive samples from CML patients who were being monitored for minimal residual disease by real-time quantitative RT-PCR were studied to determine whether the 2-DeltaDeltaCt approach was equivalent to the plasmid standard curve method. BCR/ABL P210 transcripts were quantitated using both the plasmid standard curve method and the 2-DeltaDeltaCt calculation. The comparison of both methods revealed a highly significant and linear correlation between the plasmid standard curve method and the 2-DeltaDeltaCt calculation (R2=0.98, P<0.0001). Furthermore, there was a reduction of preparation time, contamination risk, and reagent usage. The 2-DeltaDeltaCt calculation is a convenient alternative method to derive accurate quantitative information from real time PCR assays.


Ankyrin-binding proteins related to nervous system cell adhesion molecules: candidates to provide transmembrane and intercellular connections in adult brain.
A major class of ankyrin-binding glycoproteins have been identified in adult rat brain of 186, 155, and 140 kD that are alternatively spliced products of the same pre-mRNA. Characterization of cDNAs demonstrated that ankyrin-binding glycoproteins (ABGPs) share 72% amino acid sequence identity with chicken neurofascin, a membrane-spanning neural cell adhesion molecule in the Ig super-family expressed in embryonic brain. ABGP polypeptides have the following features consistent with a role as ankyrin-binding proteins in vitro and in vivo: (a) ABGPs and ankyrin associate as pure proteins in a 1:1 molar stoichiometry; (b) the ankyrin-binding site is located in the COOH-terminal 21 kD of ABGP186 which contains the predicted cytoplasmic domain; (c) ABGP186 is expressed at approximately the same levels as ankyrin (15 pmoles/milligram of membrane protein); and (d) ABGP polypeptides are co-expressed with the adult form of ankyrinB late in postnatal development and are colocalized with ankyrinB by immunofluorescence. Similarity in amino acid sequence and conservation of sites of alternative splicing indicate that genes encoding ABGPs and neurofascin share a common ancestor. However, the major differences in developmental expression reported for neurofascin in embryos versus the late postnatal expression of ABGPs suggest that ABGPs and neurofascin represent products of gene duplication events that have subsequently evolved in parallel with distinct roles. The predicted cytoplasmic domains of rat ABGPs and chicken neurofascin are nearly identical to each other and closely related to a group of nervous system cell adhesion molecules with variable extracellular domains, which includes L1, Nr-CAM, and Ng-CAM of vertebrates, and neuroglian of Drosophila. The ankyrin-binding site of rat ABGPs is localized to the C-terminal 200 residues which encompass the cytoplasmic domain, suggesting the hypothesis that ability to associate with ankyrin may be a shared feature of neurofascin and related nervous system cell adhesion molecules.


Precision measurement of the neutron twist-3 matrix element d(2)(n): probing color forces.
Double-spin asymmetries and absolute cross sections were measured at large Bjorken x (0.25≤x≤0.90), in both the deep-inelastic and resonance regions, by scattering longitudinally polarized electrons at beam energies of 4.7 and 5.9 GeV from a transversely and longitudinally polarized (3)He target. In this dedicated experiment, the spin structure function g(2)((3)He) was determined with precision at large x, and the neutron twist-3 matrix element d(2)(n) was measured at ⟨Q(2)⟩ of 3.21 and 4.32 GeV(2)/c(2), with an absolute precision of about 10(-5). Our results are found to be in agreement with lattice QCD calculations and resolve the disagreement found with previous data at ⟨Q(2)⟩=5 GeV(2)/c(2). Combining d(2)(n) and a newly extracted twist-4 matrix element f(2)(n), the average neutron color electric and magnetic forces were extracted and found to be of opposite sign and about 30 MeV/fm in magnitude.


Accuracy of after-hour 'red dot' trauma radiograph triage by radiographers in a South African regional hospital.
The global demand for diagnostic imaging exceeds the supply of radiologists and is of particular significance in poorly resourced healthcare environments where many radiographs are unreported. Delayed or absent reporting may negatively impact patient management. In well-resourced countries there is recognition that extending the role of radiographers to radiological reporting tasks helps meet service demands. Aim. To determine the accuracy of acute fracture detection by South African radiographers working in an after-hour setting. We performed a retrospective study of radiographers at a Western Cape Regional Hospital over 2 months in 2011. The sensitivity and specificity of radiographers' fracture detection were compared with that of a consultant radiologist. Differences were evaluated using the McNemar chi-squared test, with p<0.05 regarded as significant. A total of 369 radiographs were analysed. The overall accuracy of reporting by radiographers was 93.7%, with 74.4% sensitivity for fracture detection. Experienced radiographers performed better than inexperienced radiographers; adult fractures were more consistently identified than paediatric fractures, and appendicular fractures were better visualised than axial fractures. In all instances there was a significant difference between fracture detection by radiographers and the radiologist. Experienced radiographers evaluating appendicular fractures in adults achieved the highest sensitivity (89.9%), which was not significantly different from that of a consultant radiologist (p=0.88). The performance of experienced radiographers in our study is comparable with that of experienced radiographers internationally, who have no specific training in trauma radiograph reporting. However, additional training is required if role extension is to be considered.


Solution-processed two-dimensional layered heterostructure thin-film with optimized thermoelectric performance.
Graphene-based two-dimensional (2D) heterostructures have ignited intensive interest in recent years because of their excellent physical performance. However, the most common method to prepare them uses chemical vapor deposition, which has the drawback of a complex process unsuitable for large-scale production. In this respect, reduced graphene oxide and transitional metal dichalcogenides (rGO-TMDs) composite thin-films were fabricated by a simple solution-processing method and their thermoelectric performance was investigated systematically. Addition of rGO nanosheets (NSs) efficiently improved the electrical conductivity of MoS2 and WS2 (MS2) NSs, due to the excellent electron transport performance of rGO. Furthermore, it should be noted that an optimized content of rGO can effectively avoid direct contact between TMDs NSs by forming a rGO-TMDs heterojunction, leading to significantly increased electrical conductivity and a slight variation in its Seebeck coefficient. Our work obtained high thermoelectric performance heterostructures by inducing two kinds of layered materials using a simple method that may potentially be applied to other 2D layered materials to construct heterostructures for energy conversion.


The tripartite type III secreton of Shigella flexneri inserts IpaB and IpaC into host membranes.
Bacterial type III secretion systems serve to translocate proteins into eukaryotic cells, requiring a secreton and a translocator for proteins to pass the bacterial and host membranes. We used the contact hemolytic activity of Shigella flexneri to investigate its putative translocator. Hemolysis was caused by formation of a 25-A pore within the red blood cell (RBC) membrane. Of the five proteins secreted by Shigella upon activation of its type III secretion system, only the hydrophobic IpaB and IpaC were tightly associated with RBC membranes isolated after hemolysis. Ipa protein secretion and hemolysis were kinetically coupled processes. However, Ipa protein secretion in the immediate vicinity of RBCs was not sufficient to cause hemolysis in the absence of centrifugation. Centrifugation reduced the distance between bacterial and RBC membranes beyond a critical threshold. Electron microscopy analysis indicated that secretons were constitutively assembled at 37 degrees C before any host contact. They were composed of three parts: (a) an external needle, (b) a neck domain, and (c) a large proximal bulb. Secreton morphology did not change upon activation of secretion. In mutants of some genes encoding the secretion machinery the organelle was absent, whereas ipaB and ipaC mutants displayed normal secretons.


Release of the export adapter, Nmd3p, from the 60S ribosomal subunit requires Rpl10p and the cytoplasmic GTPase Lsg1p.
In eukaryotes, nuclear export of the large (60S) ribosomal subunit requires the adapter protein Nmd3p to provide the nuclear export signal. Here, we show that in yeast release of Nmd3p from 60S subunits in the cytoplasm requires the ribosomal protein Rpl10p and the G-protein, Lsg1p. Mutations in LSG1 or RPL10 blocked Nmd3-GFP shuttling into the nucleus and export of pre-60S subunits from the nucleus. Overexpression of NMD3 alleviated the export defect, indicating that the block in 60S export in lsg1 and rpl10 mutants results indirectly from failing to recycle Nmd3p. The defect in Nmd3p recycling and the block in 60S export in both lsg1 and rpl10 mutants was also suppressed by mutant Nmd3 proteins that showed reduced binding to 60S subunits in vitro. We propose that the correct loading of Rpl10p into 60S subunits is required for the release of Nmd3p from subunits by Lsg1p. These results suggest a coupling between recycling the 60S export adapter and activation of 60S subunits for translation.


Granular topological insulators.
We demonstrate experimentally that a macroscopic topological insulator (TI) phase can emerge in a granular conductor composed of an assembly of tunnel coupled TI nanocrystals of dimension ∼10 nm × 10 nm × 2 nm. Electrical transport measurements on thin films of Bi2Se3 nanocrystals reveal the presence of decoupled top and bottom topological surface states that exhibit large surface state penetration depths (∼30 nm at 2 K). By tuning the size of the nanocrystals and the couplings between them, this new class of TIs may be readily tuned from a non-topological to a topological insulator phase, that too with designer properties. Paradoxically, this seemingly 'dirty' system displays properties that are closer to an ideal TI than most known single crystal systems, making granular/nanocrystalline TIs an attractive platform for future TI research.


Differences in mutagenicity and cytotoxicity of (+)- and (-)-benzo[a]pyrene 4,5-oxide: a synergistic interaction of enantiomers.
In order to study the biological effects of (+)- and (-)-benzo[a]pyrene 4,5-oxide, a synthesis of these molecules has been developed based on the resolution of (+/-)-cis-4,5-dihydroxy-4,5-dihydrobenzo[a]pyrene. The (-) enantiomer of benzo[a]pyrene 4,5-oxide was 1.5- to 5.5-fold more mutagenic than the (+) enantiomer in strains TA 98, TA 100, TA 1537, and TA 1538 of Salmonella typhimurium and in Chinese hamster V79 cells. In studies with V79 Cells, the (-) enantiomer of benzo[a]pyrene 4,5-oxide was also more cytotoxic than the (+) enantiomer. When mixtures of the enantiomers were studied in V79 cells, synergistic cytotoxic and mutagenic responses were observed. The greatest cytotoxic and mutagenic effects occurred with a 3:1 mixture of the (-) and (+) enantiomers of benzo[a]pyrene 4,5-oxide, respectively.


Serum ficolin-2 correlates worse than fecal calprotectin and CRP with endoscopic Crohn's disease activity.
Ficolin-2 is an acute phase reactant produced by the liver and targeted to recognize N-acetyl-glucosamine which is present in bacterial and fungal cell walls. We recently showed that ficolin-2 serum levels were significantly higher in CD patients compared to healthy controls. We aimed to evaluate serum ficolin-2 concentrations in CD patients regarding their correlation with endoscopic severity and to compare them with clinical activity, fecal calprotectin, and CRP. Patients provided fecal and blood samples before undergoing ileo-colonoscopy. Disease activity was scored clinically according to the Harvey-Bradshaw Index (HBI) and endoscopically according to the simplified endoscopic score for CD (SES-CD). Ficolin-2 serum levels and fecal calprotectin levels were measured by ELISA. A total of 136 CD patients were prospectively included (mean age at inclusion 41.5±15.4 years, 37.5% females). Median HBI was 3 [2-6] points, median SES-CD was 5 [2-8], median fecal calprotectin was 301 [120-703] μg/g, and median serum ficolin-2 was 2.69 [2.02-3.83] μg/mL. SES-CD correlated significantly with calprotectin (R=0.676, P<0.001), CRP (R=0.458, P<0.001), HBI (R=0.385, P<0.001), and serum ficolin-2 levels (R=0.171, P=0.047). Ficolin-2 levels were higher in CD patients with mild endoscopic disease compared to patients in endoscopic remission (P=0.015) but no difference was found between patients with mild, moderate, and severe endoscopic disease. Ficolin-2 serum levels correlate worse with endoscopic CD activity when compared to fecal calprotectin or CRP.


Effects of a simulated wolf encounter on brain and blood biomarkers of stress-related psychological disorders in beef cows with or without previous exposure to wolves.
This experiment compared mRNA expression of brain-blood biomarkers associated with stress-related psychological disorders, including post-traumatic stress disorder (PTSD), in beef cows from wolf-naïve and wolf-experienced origins that were subjected to a simulated wolf encounter. Multiparous, non-pregnant, non-lactating Angus-crossbred cows from the Eastern Oregon Agricultural Research Center (Burns, OR; CON; = 10) and from a commercial operation near Council, ID (WLF; = 10) were used. To date, gray wolves are not present around Burns, OR, and thus CON were naïve to wolves. Conversely, wolves are present around Council, ID, and WLF cows were selected from a herd that had experienced multiple wolf-predation episodes from 2008 to 2015. After a 60-d commingling and adaptation period, CON and WLF cows were allocated to groups A or B (d -1; 5 CON and 5 WLF cows in each group). On d 0, cows from group A were sampled for blood and immediately slaughtered, and samples were analyzed to evaluate inherent differences between CON and WLF cows. On d 1, cows from group B were exposed in pairs (1 CON and 1 WLF cow) to experimental procedures. Cows were sampled for blood, moved to 2 adjacent drylot pens (1 WLF and 1 CON cow/pen) and subjected to a simulated wolf encounter event for 20 min. The encounter consisted of (1) cotton plugs saturated with wolf urine attached to the drylot fence, (2) reproduction of wolf howls, and (3) three leashed dogs that were walked along the fence perimeter. Thereafter, another blood sample was collected and cows were slaughtered. Upon slaughter, the brain was removed and dissected for collection of the hypothalamus, and one longitudinal slice of the medial pre-frontal cortex, amygdala, and Cornu Ammonis (1 region of the hippocampus from both hemispheres). Within cows from group A, expression of in hippocampus and amygdala were greater ( < 0.01) in WLF vs. CON cows. Within cows from group B, expression of hippocampal mRNA and expression of c mRNA in hippocampus and amygdala were less ( ≤ 0.04) in WLF vs. CON cows. These are key biological markers known to be downregulated during stress-related psychological disorders elicited by fear, particularly PTSD. Hence, cows originated from a wolf-experienced herd presented biological evidence suggesting a psychological disorder, such as PTSD, after the simulated wolf encounter when compared with cows originated from a wolf-naïve herd.


The Ameliorating Effect of Myrrh on Scopolamine-Induced Memory Impairments in Mice.
Myrrh has been used since ancient times for the treatment of various diseases such as inflammatory diseases, gynecological diseases, and hemiplegia. In the present study, we investigated the effects of aqueous extracts of myrrh resin (AEM) on scopolamine-induced memory impairments in mice. AEM was estimated with (2E,5E)-6-hydroxy-2,6-dimethylhepta-2,4-dienal as a representative constituent by HPLC. The oral administration of AEM for 7 days significantly reversed scopolamine-induced reduction of spontaneous alternation in the Y-maze test. In the passive avoidance task, AEM also restored the decreased latency time of the retention trial by scopolamine treatment. In addition, Western blot analysis and Immunohistochemistry revealed that AEM reversed scopolamine-decreased phosphorylation of Akt and extracellular signal-regulated kinase (ERK). Our study demonstrates for the first time that AEM ameliorates the scopolamine-induced memory impairments in mice and increases the phosphorylation of Akt and ERK in the hippocampus of mice brain. These results suggest that AEM has the therapeutic potential in memory impairments.


Maturation of 6S regulatory RNA to a highly elongated structure.
As bacterial populations leave the exponential growth phase and enter the stationary phase, their patterns of gene expression undergo marked changes. A key effector of this change is 6S RNA, which is a highly conserved regulatory RNA that impedes the transcription of genes associated with exponential growth by forming an inactivating ternary complex with RNA polymerase and sigma factor σ(70) (σ(70)-RNAP). In Escherichia coli, the endoribonuclease RNase E generates 6S RNA by specific cleavage of a precursor that is nearly twice the size of the 58 kDa mature form. We have explored recognition of the precursor by RNase E, and observed that processing is inhibited under conditions of excess substrate. This finding supports a largely distributive mechanism, meaning that each round of catalysis results in enzyme dissociation and re-binding to the substrate. We show that the precursor molecule and the mature 6S share a structural core dominated by an A-type helix, indicating that processing is not accompanied by extensive refolding. Both precursor and mature forms of 6S have a highly stable secondary structure, adopt an elongated shape, and show the potential to form dimers under specific conditions; nonetheless, 6S has a high structural plasticity that probably enables it to be structurally adapted upon binding to its cognate protein partners. Analysis of the 6S-σ(70)-RNAP complex by native mass spectrometry reveals a stable association with a stoichiometry of 1:1:1. A theoretical 3D model of mature 6S is presented, which is consistent with the experimental data and supports a previously proposed structure with a small stem-loop inside the central bubble.


Nonalcoholic fatty liver disease can be predicted by retinal vascular changes in patients with obesity without hypertension or diabetes.
To evaluate the utility of predicting nonalcoholic fatty liver disease (NAFLD) and obesity by retinal vascular changes (RVC) found in fundoscopy and to determine whether this is related to a low-grade inflammatory process. We carried out a cross-sectional analysis that included 152 (ages 18-45 years) patients divided into four groups: NAFLD and BMI less than 25, absence of NAFLD and BMI less than 25, NAFLD and BMI more than 30, and absence of NAFLD and BMI more than 30. Retinal fundoscopy, hepatic ultrasound, metabolic profile, and cytokine measurement were performed. We calculated odds ratio [95% confidence interval (CI)], performed diagnostic utility tests, and carried out a 2 factorial analysis. Obesity was associated with RVC (odds ratio: 21.25; 95% CI: 8.79-51.4) and NAFLD [25 (9.07-72.6)]. NAFLD was associated with RVC [11.24 (4.98-26.48)], and the prediction of NAFLD showed a sensitivity of 75% (95% CI: 68-82) and a specificity of 81% (75-86); when RVC-obesity were combined, sensitivity increased to 90% (88-91.7), with a specificity of 85% (84-85.8). C-reactive protein was associated with the three factors, suggesting an independent contribution. Thin patients with RVC and NAFLD had higher concentrations of interleukin-2, interleukin-6, tumor necrosis factor-α, and interferon-γ. NAFLD in patients with obesity without diabetes or hypertension can be predicted by RVC, a noninvasive technique carried out by eye fundoscopy. NAFLD alone can drive inflammatory conditions in the absence of obesity that manifests as RVC.


Sighting acute myocardial infarction through platelet gene expression.
Acute myocardial infarction is primarily due to coronary atherosclerotic plaque rupture and subsequent thrombus formation. Platelets play a key role in the genesis and progression of both atherosclerosis and thrombosis. Since platelets are anuclear cells that inherit their mRNA from megakaryocyte precursors and maintain it unchanged during their life span, gene expression profiling at the time of an acute myocardial infarction provides information concerning the platelet gene expression preceding the coronary event. In ST-segment elevation myocardial infarction (STEMI), a gene-by-gene analysis of the platelet gene expression identified five differentially expressed genes: FKBP5, S100P, SAMSN1, CLEC4E and S100A12. The logistic regression model used to combine the gene expression in a STEMI vs healthy donors score showed an AUC of 0.95. The same five differentially expressed genes were externally validated using platelet gene expression data from patients with coronary atherosclerosis but without thrombosis. Platelet gene expression profile highlights five genes able to identify STEMI patients and to discriminate them in the background of atherosclerosis. Consequently, early signals of an imminent acute myocardial infarction are likely to be found by platelet gene expression profiling before the infarction occurs.


Treatment of nonexudative (dry) age-related macular degeneration.
The purpose of this report is to review the recent literature and summarize currently available and potential new treatment options for nonexudative age-related macular degeneration. High-dose vitamin supplementation may have some associated systemic toxicity. It is important to check that the patient is taking beta-carotene and not vitamin A as retinal acetate or palmitate, which have been associated with osteoporosis and hepatotoxicity. High-dose vitamins E and C may be associated with cardiovascular disease. Decreasing inflammation by lowering systemic cardiac C-reactive protein, fibrinogen and cholesterol may be important, especially in light of recent epidemiologic and genetic data. The results of randomized trials of laser treatment for drusen and rheopheresis should be available during 2006. Treatment with these modalities before the results of the trials are evaluated should be avoided. The holy grail of therapy for age-related macular degeneration is to avoid the development of choroidal neovascularization. High-dose vitamin supplementation should be used only in those in whom it is indicated and inflammatory parameters including highly sensitive C-reactive protein, fibrinogen and cholesterol should be stabilized because there are data associating these parameters with age-related macular degeneration and also with cardiovascular disease.


Effect of increased blood levels of β-endorphin on perception of breathlessness.
Although opioid receptors are expressed broadly in the CNS and in peripheral sensory nerve endings including bronchioles and alveolar walls of the respiratory tract, it is unknown whether the modulatory effect of endogenous opioids on breathlessness occurs in the CNS or in the peripheral nervous system. The purpose of this investigation was to examine whether increased blood levels of β-endorphin modify breathlessness by a putative effect of binding to peripheral opioid receptors in the respiratory tract. Twenty patients with COPD (10 women and 10 men; age, 70 ± 8 years) inspired through resistances during practice sessions to identify an individualized target load that caused ratings of intensity and unpleasantness of breathlessness ≥ 50 mm on a 100-mm visual analog scale. At two interventions, blood levels of β-endorphin and adrenocorticotropic hormone (ACTH) were measured, ketoconazole (600 mg) or placebo was administered orally, and patients rated the two dimensions of breathlessness each minute during resistive load breathing (RLB). By inhibiting cortisol synthesis, ketoconazole led to significant increases in β-endorphin (mean change, 20% ± 4%) and ACTH (mean change, 21% ± 4%) compared with placebo. The intensity and unpleasantness ratings of breathlessness and the endurance time during RLB were similar in the two interventions. The previously demonstrated modulatory effect of endogenous opioids on breathlessness appears to be mediated by binding to receptors within the CNS rather than to peripheral opioid receptors in the respiratory tract. An alternative explanation is that the magnitude of the β-endorphin response is inadequate to affect peripheral opioid receptors. ClinicalTrials.gov; No.: NCT01378520; URL: www.clinicaltrials.gov.


Multiple levels of influence on older adults' attendance and adherence to community exercise classes.
To examine the influence of individual participant, instructor, and group factors on participants' attendance and adherence to community exercise classes for older adults. Longitudinal data from 16 instructors, 26 classes, and 193 older participants within those classes (aged 60-100 years) were examined. Data were collected using questionnaires on individual participants' demographics, attitudes, health perceptions and conditions, and group cohesion. Instructors' demographics, training, background, experience, attitudes, and personality were collected. Group factors included class type, cost, transport, and whether the class was held in an area of deprivation. Outcomes (attendance/adherence) were collected through attendance records. Multilevel modelling (MLwiN) revealed both instructor and individual participant variables were important in understanding attendance and adherence. Individuals' housing, education, mental well-being, group cohesion, and attitudes were important predictors of attendance at 3 and 6 months. Instructors' age, gender, experience, and motivational training were important at 3 months, whereas instructor personality was important at both 3 and 6 months. Having attended longer than 6 months at baseline, participants' attitudes, weeks offered, instructors' personality, and experience were associated with adherence at 6 months. Results suggest that instructors' characteristics alongside individual participant factors play a role in influencing participants' attendance to exercise classes. These factors should be considered when setting up new programs.


How long does an elbow replacement last? A systematic review and meta-analysis of case-series and national registry reports with more than 10 years of follow-up.
This study aims to determine, for the first time, generalizable data on the longevity and long-term function of elbow replacements. In this systematic review and meta-analysis, we searched MEDLINE and Embase for articles reporting 10-year or greater survival of total elbow replacements (TERs) and distal humeral hemiarthroplasty. Implant survival and patient reported outcome measures (PROMs) data were extracted. National joint replacement registries were also analyzed. We weighted each series and calculated a pooled survival estimate at 10, 15, and 20 years. For PROMs we pooled the standardized mean difference (SMD) at 10 years. Despite its widespread use, we identified only 9 series reporting all-cause survival of 628 linked TERs and 610 unlinked TERs and no series for distal humeral hemiarthroplasty. The studied population was treated for rheumatoid arthritis in over 90% of cases. The estimated 10-year survival for linked TERs was 92% (95% CI 90-95) and unlinked TERs 84% (CI 81-88). 2 independent registries contributed 32 linked TERs and 530 unlinked TERs. The pooled registry 10-year survival for unlinked TERs was 86% (CI 83-89). Pooled 10-year PROMs from 164 TERs (33 linked and 131 unlinked), revealed a substantial improvement from baseline scores (SMD 2.7 [CI 1.6-3.8]). Over 80% of all elbow replacements and over 90% of linked elbow replacements can last more than 10 years with sustained patient-reported benefits. This information is long overdue and will be particularly useful to patients as well as healthcare providers.


Information presentation through a head-worn display ("smart glasses") has a smaller influence on the temporal structure of gait variability during dual-task gait compared to handheld displays (paper-based system and smartphone).
The need to complete multiple tasks concurrently is a common occurrence both daily life and in occupational activities, which can often include simultaneous cognitive and physical demands. As one example, there is increasing availability of head-worn display technologies that can be employed when a user is mobile (e.g., while walking). This new method of information presentation may, however, introduce risks of adverse outcomes such as a decrement to gait performance. The goal of this study was thus to quantify the effects of a head-worn display (i.e., smart glasses) on motor variability during gait and to compare these effects with those of other common information displays (i.e., smartphone and paper-based system). Twenty participants completed four walking conditions, as a single task and in three dual-task conditions (three information displays). In the dual-task conditions, the information display was used to present several cognitive tasks. Three different measures were used to quantify variability in gait parameters for each walking condition (using the cycle-to-cycle standard deviation, sample entropy, and the "goal-equivalent manifold" approach). Our results indicated that participants used less adaptable gait strategies in dual-task walking using the paper-based system and smartphone conditions compared with single-task walking. Gait performance, however, was less affected during dual-task walking with the smart glasses. We conclude that the risk of an adverse gait event (e.g., a fall) in head-down walking conditions (i.e., the paper-based system and smartphone conditions) were higher than in single-task walking, and that head-worn displays might help reduce the risk of such events during dual-task gait conditions.


Salivary cortisol during memory encoding in pregnancy predicts postpartum depressive symptoms: a longitudinal study.
Postpartum depression (PPD) is a common disorder that substantially decreases quality of life for both mother and child. In this longitudinal study, we investigated whether emotional memory, salivary cortisol (sCORT) or alpha-amylase during pregnancy predict postpartum depressive symptoms. Forty-four pregnant women (14 euthymic women with a diagnosis of major depressive disorder [MDD] and 30 healthy women) between the ages of 19 and 37 years (mean age = 29.5±4.1 years) were longitudinally assessed in the 2nd trimester of pregnancy (12-22 weeks of gestational age) and again at 14-17 weeks postpartum. Depressive symptoms were assessed using the Edinburgh Postnatal Depression Scale (EPDS). Follow-ups were completed for 41 women (7% attrition). Postpartum EPDS scores were predicted by sCORT collected immediately after an incidental encoding memory task during pregnancy (b=-0.78, t -2.14, p=0.04). Postpartum EPDS scores were not predicted by positive (p=0.27) or negative (p=0.85) emotional memory. The results of this study indicate that higher levels of sCORT during a memory encoding task in the 2nd trimester of pregnancy are associated with lower postpartum EPDS scores. While the hypothalamus-pituitary-adrenal (HPA) axis has long been associated with the neurobiology of MDD, the role of the HPA axis in perinatal depression deserves more attention.


Mutations in JARID1C are associated with X-linked mental retardation, short stature and hyperreflexia.
Mutations in the JARID1C (Jumonji AT-rich interactive domain 1C) gene were recently associated with X-linked mental retardation (XLMR). Mutations in this gene are reported to be one of the relatively more common causes of XLMR with a frequency of approximately 3% in males with proven or probable XLMR. The JARID1C protein functions as a histone 3 lysine 4 (H3K4) demethylase and is involved in the demethylation of H3K4me3 and H3K4me2. Mutation analysis of the JARID1C gene was conducted in the following cohorts: probands from 23 XLMR families linked to Xp11.2, 92 males with mental retardation and short stature, and 172 probands from small XLMR families with no linkage information. Four novel mutations consisting of two missense mutations, p.A77T and p.V504M, and two frame shift mutations, p.E468fsX2 and p.R1481fsX9, were identified in males with mental retardation. Two of the mutations, p.V504M and p.E468fsX2, are located in the JmjC domain of the JARID1C gene where no previous mutations have been reported. Additional studies showed that the missense mutation, p.V504M, was a de novo event on the grandpaternal X chromosome of the family. Clinical findings of the nine affected males from the four different families included mental retardation (100%), short stature (55%), hyperreflexia (78%), seizures (33%) and aggressive behaviour (44%). The degree of mental retardation consisted of mild (25%), moderate (12%) and severe (63%). Based on the clinical observations, male patients with mental retardation, short stature and hyperreflexia should be considered candidates for mutations in the JARID1C gene.


MicroRNA-31 predicts the presence of lymph node metastases and survival in patients with lung adenocarcinoma.
We conducted genome-wide miRNA-sequencing (miRNA-seq) in primary cancer tissue from patients of lung adenocarcinoma to identify markers for the presence of lymph node metastasis. Markers for lymph node metastasis identified by sequencing were validated in a separate cohort using quantitative PCR. After additional validation in the The Cancer Genome Atlas (TCGA) dataset, functional characterization studies were conducted in vitro. MiR-31 was upregulated in lung adenocarcinoma tissues from patients with lymph node metastases compared with those without lymph node metastases. We confirmed miR-31 to be upregulated in lymph node-positive patients in a separate patient cohort (P = 0.009, t test), and to be expressed at higher levels in adenocarcinoma tissue than in matched normal adjacent lung tissues (P < 0.0001, paired t test). MiR-31 was then validated as a marker for lymph node metastasis in an external validation cohort of 233 lung adenocarcinoma cases of the TCGA (P = 0.031, t test). In vitro functional assays showed that miR-31 increases cell migration, invasion, and proliferation in an ERK1/2 signaling-dependent manner. Notably, miR-31 was a significant predictor of survival in a multivariate cox regression model even when controlling for cancer staging. Exploratory in silico analysis showed that low expression of miR-31 is associated with excellent survival for T2N0 patients. We applied miRNA-seq to study microRNomes in lung adenocarcinoma tissue samples for the first time and potentially identified a miRNA predicting the presence of lymph node metastasis and survival outcomes in patients of lung adenocarcinoma.


Perceived glossiness in high dynamic range scenes.
We investigated how spatial pattern, background, and dynamic range affect perceived gloss in brightly lit real scenes. Observers viewed spherical objects against uniform backgrounds. There were three possible objects. Two were black matte spheres with circular matte white dots painted on them (matte-dot spheres). The third sphere was painted glossy black (glossy black sphere). Backgrounds were either black or white matte, and observers saw each of the objects in turn on each background. Scenes were illuminated by an intense collimated source. On each trial, observers matched the apparent albedo of the sphere to an albedo reference scale and its apparent gloss to a gloss reference scale. We found that matte-dot spheres and the black glossy sphere were perceived as glossy on both backgrounds. All spheres were judged to be significantly glossier when in front of the black background. In contrast with previous research using conventional computer displays, we find that background markedly affects perceived gloss. This finding is surprising because darker surfaces are normally perceived as glossier (F. Pellacini, J. A. Ferwerda, & D. P. Greenberg, 2000). We conjecture that there are cues to surface material signaling glossiness present in high dynamic range scenes that are absent or weak in scenes presented using conventional computer displays.


Characterization of gastrins and their receptor in solid human gastric adenocarcinomas.
The gastrin and the gastrin/CCK-B receptor genes are co-expressed in several carcinomas. The primary translational product, progastrin, however, is processed to several peptides of which only those that are α-amidated at their C-terminus are receptor ligands. So far, characterization of the progastrin-derived peptides in gastric cancer has not been reported. The authors therefore examined the molecular nature of gastrin and its receptor in human gastric carcinomas. Twenty patients with adenocarcinoma underwent partial or total gastrectomy. In samples from each carcinoma, gastrin peptides were characterized, using a library of sequence-specific immunoassays. Expression was also demonstrated by immunohistochemistry. In addition, the gastrin and gastrin/CCK-B receptor gene expression was quantitated using real-time PCR, and the receptor protein demonstrated by western blotting. α-Amidated gastrins were detectable in 16 of 20 carcinomas (median concentration 2.1 pmol/g tissue; range 0-386 pmol/g tissue). The tissue concentrations correlated closely to the gastrin mRNA contents (r = 0.75, p < 0.0001). Moreover, progastrin and non-amidated processing intermediates, including glycine-extended gastrins, were detected in 19 carcinomas. Immunohistochemistry corroborated gastrin expression in carcinoma cells. Chromatography revealed extensive progastrin processing with α-amidated gastrin-34 and -17 (tyrosyl-sulfated as well as non-sulfated) as major products. Finally, gastrin/CCK-B receptor mRNA and protein were detected in all tumors. The results show that the elements for a local loop of α-amidated gastrins and their receptor are detectable in 80% of human gastric adenocarcinomas. Therefore, the results support the contention that locally expressed gastrin may be involved in the tumorigenesis of gastric adenocarcinomas.


New connections in the regulation of PEPCK gene expression by insulin.
Phosphoenolpyruvate carboxykinase (PEPCK) catalyses the rate-limiting step in hepatic gluconeogenesis. Glucagon (via the second messenger cAMP) and glucocorticoids stimulate transcription of the PEPCK gene whereas insulin and phorbol esters have a dominant inhibitory effect. Wortmannin, an inhibitor of 1-phosphatidylinositol 3-kinase (PI 3-kinase), blocks the inhibition of glucocorticoid- and cAMP-stimulated PEPCK gene transcription by insulin. By contrast, although phorbol esters mimic the action of insulin on the regulation of PEPCK gene transcription, wortmannin does not block the effect of these agents. Thus PI 3-kinase is required for the regulation of PEPCK gene expression by insulin but not by phorbol esters. In liver cells, insulin administration stimulates the activity of multiple protein kinases, including the p42/p44 Mitogen Activated Protein (MAP) kinase and the p70/p85 ribosomal protein S6 kinase. Selective inhibition of the activation of either kinase, utilizing the compounds PD98059 and rapamycin respectively, does not affect insulin regulation of PEPCK gene transcription. Thus regulation of PEPCK gene transcription requires PI 3-kinase but does not require the activation of either p42/p44 MAP kinase or p70/p85 ribosomal protein S6 kinase.


Clinical research: remote magnetic navigation vs. manually controlled catheter ablation of right ventricular outflow tract arrhythmias: a retrospective study.
Remote magnetic navigation (RMN) is an alternative to manual catheter control (MCC) radiofrequency ablation of right ventricular outflow tract (RVOT) arrhythmias. The data to support RMN approach is limited. We aimed to investigate the clinical and procedural outcomes in a cohort of patients undergoing RVOT premature ventricular complex/ventricular tachycardia (PVCs/VT) ablation procedures using RMN vs. MCC. Data was collected from two centres. Eighty-nine consecutive RVOT PVCs/VT ablation procedures were performed in 75 patients; RMN: 42 procedures and MCC: 47 procedures. CARTOXPTM or CARTO3 (Biosense Webster) was used for endocardial mapping in 19/42 (45%) in RMN group and 28/47 (60%) in MCC group; EnSiteTM NavXTM (St. Jude Medical) was used in the rest of the cohort. Stereotaxis platform (Stereotaxis Inc., St. Louis, MO, USA) was used for RMN approach. Procedural time was 113 ± 53 min in the RMN group and 115 ± 69 min in MCC (P = 0.90). Total fluoroscopic time was 10.9 ± 5.8 vs. 20.5 ± 13.8 (P < 0.05) and total ablation energy application time 7.0 ± 4.7 vs 11.9 ± 16 (P = 0.67) accordingly. There were two complications in RMN group and five in MCC (P = 0.43). Acute procedural success rate was 80% in RMN vs. 74% in MCC group (P = 0.46). After a median follow-up of 25 months (interquartile range 13-34), the success rate remained 55% in the RMN group and 53% in MCC (P = 0.96). Right ventricular outflow tract arrhythmia ablations were performed using half of fluoroscopic times with Stereotaxis platform RMN compared to manual approach. Acute and chronic success rates as well as complication rates were not significantly different.


Characteristics of Epstein-Barr virus activation of human B lymphocytes.
Epstein-Barr virus (EBV) will infect at least every third cell if exposed in vitro to an extensively purified B cell population from human peripheral blood. About 10% of such infected cells will be driven into immunoglobulin synthesis and secretion, as judged by the indirect protein A plaque assay. The appearance of EB nuclear antigen, de novo DNA synthesis, and immunoglobulin secretion are linked phenomena accompanying infection as judged by viral dilution experiments, which yield kinetics of a one-hit order. Induction of immunoglobulin secretion in B cells by EBV requires de novo synthesis of DNA, and consequently, nontransforming EBV (P3HR1) will not induce immunoglobulin secretion and will also specifically block such induction from subsequently added EBV. The termination of immunoglobulin induction by EBV in short-term cultures appears to be T cell dependent.


The association of complex liver disorders with HBV genotypes prevalent in Pakistan.
Genotyping of HBV is generally used for determining the epidemiological relationship between various virus strains and origin of infection mostly in research studies. The utility of genotyping for clinical applications is only beginning to gain importance. Whether HBV genotyping will constitute part of the clinical evaluation of Hepatitis B patients depends largely on the availability of the relevance of the evidence based information. Since Pakistan has a HBV genotype distribution which has been considered less virulent as investigated by earlier studies from south East Asian countries, a study on correlation between HBV genotypes and risk of progression to further complex hepatic infection was much needed A total of 295 patients with HBsAg positive were selected from the Pakistan Medical Research Council's (PMRC) out patient clinics. Two hundred and twenty six (77%) were males, sixty nine (23%) were females (M to F ratio 3.3:1). Out of 295 patients, 156 (53.2%) had Acute(CAH), 71 (24.2%) were HBV Carriers, 54 (18.4%) had Chronic liver disease (CLD) Hepatitis. 14 (4.7%) were Cirrhosis and HCC patients. Genotype D was the most prevalent genotype in all categories of HBV patients, Acute (108), Chronic (39), and Carrier (53).Cirrhosis/HCC (7) were HBV/D positive. Genotype A was the second most prevalent with 28 (13%) in acute cases, 12 (22.2%) in chronics, 14 (19.7%) in carriers and 5 (41.7) in Cirrhosis/HCC patients. Mixed genotype (A/D) was found in 20 (12.8%) of Acute patients, 3 (5.6%) of Chronic and 4 (5.6%) of carriers, none in case of severe liver conditions. Mixed HBV genotypes A, D and A/D combination were present in all categories of patients except that no A/D combination was detected in severe conditions. Genotype D was the dominant genotype. However, genotype A was found to be more strongly associated with severe liver disease. Mixed genotype (A/D) did not significantly appear to influence the clinical outcome.


Does the use of ingredients added to tobacco increase cigarette addictiveness?: a detailed analysis.
The possibility that ingredients added to tobacco contribute to the addictiveness of cigarette smoking was evaluated by comparing cessation rates of smokers of traditional blended cigarettes to those of smokers of flue-cured cigarettes. Such a comparison is a valid means of assessing cigarette ingredients as traditional blended cigarettes contain ingredients (>20), whereas flue-cured cigarettes contain no or very few ingredients. Separate analysis of 108 treatment groups and 108 control groups from randomized clinical trials of nicotine replacement therapy (NRT) were performed by multiple logistic regressions. The results of these analyses demonstrated slightly higher quit rates for smokers of blended cigarettes (OR = 1.90, 95% CI 1.70-2.13 and OR = 1.32, 95% CI 1.14-1.53 for treatment and control groups, respectively). The control groups were also investigated using classification tree analysis from which no difference in quit rates were observed for smokers of either type of cigarette. Further analyses showed that studies that utilized a high level of psychological support in conjunction with NRT produced at least a two-fold increase in quit rates compared to studies that utilized a low level of psychological support. It was also demonstrated that there is a large difference when results were reported by sustained abstinence compared to point prevalence. Additional meta-analyses found the pooled OR for NRT treatment to be in exact agreement with a recent review that assessed the effectiveness of NRT. Overall these results strongly suggest that ingredients used in the manufacture of traditional blended cigarettes do not increase the inherent addictiveness of cigarettes.


The CYB5R3c .350C>G and G6PD A alleles modify severity of anemia in malaria and sickle cell disease.
Genetic modifiers of anemia in Plasmodium falciparum infection and sickle cell disease (SCD) are not fully known. Both conditions are associated with oxidative stress, hemolysis and anemia. The CYB5R3 gene encodes cytochrome b5 reductase 3, which converts methemoglobin to hemoglobin through oxidation of NADH. CYB5R3c.350C > G encoding CYB5R3T117S , the most frequent recognized African-specific polymorphism, does not have known functional significance, but its high allele frequency (23% in African Americans) suggests a selection advantage. Glucose-6-phosphate dehydrogenase (G6PD) is essential for protection from oxidants; its African-polymorphic X-linked A+ and A- alleles, and other variants with reduced activity, coincide with endemic malaria distribution, suggesting protection from lethal infection. We examined the association of CYB5R3c.350C > G with severe anemia (hemoglobin <5 g/dL) in the context of G6PD A+ and A- status among 165 Zambian children with malaria. CYB5R3c.350C > G offered protection against severe malarial anemia in children without G6PD deficiency (G6PD wild type or A+/A- heterozygotes) (odds ratio 0.29, P = .022) but not in G6PD A+ or A- hemizygotes/homozygotes. We also examined the relationship of CYB5R3c.350C > G with hemoglobin concentration among 267 children and 321 adults and adolescents with SCD in the US and UK and found higher hemoglobin in SCD patients without G6PD deficiency (β = 0.29, P = .022 children; β = 0.33, P = .004 adults). Functional studies in SCD erythrocytes revealed mildly lower activity of native CYB5R3T117S compared to wildtype CYB5R3 and higher NADH/NAD+ ratios. In conclusion, CYB5R3c.350C > G appears to ameliorate anemia severity in malaria and SCD patients without G6PD deficiency, possibly accounting for CYB5R3c.350C > G selection and its high prevalence.


Dissemination of Mycobacterium tuberculosis is associated to a SIGLEC1 null variant that limits antigen exchange via trafficking extracellular vesicles.
The identification of individuals with null alleles enables studying how the loss of gene function affects infection. We previously described a non-functional variant in SIGLEC1, which encodes the myeloid-cell receptor Siglec-1/CD169 implicated in HIV-1 cell-to-cell transmission. Here we report a significant association between the SIGLEC1 null variant and extrapulmonary dissemination of Mycobacterium tuberculosis (Mtb) in two clinical cohorts comprising 6,256 individuals. Local spread of bacteria within the lung is apparent in Mtb-infected Siglec-1 knockout mice which, despite having similar bacterial load, developed more extensive lesions compared to wild type mice. We find that Siglec-1 is necessary to induce antigen presentation through extracellular vesicle uptake. We postulate that lack of Siglec-1 delays the onset of protective immunity against Mtb by limiting antigen exchange via extracellular vesicles, allowing for an early local spread of mycobacteria that increases the risk for extrapulmonary dissemination.


Risk factors for advanced liver fibrosis in HIV-infected individuals: role of antiretroviral drugs and insulin resistance.
Liver damage may result from multiple factors in HIV-infected patients. The availability of reliable noninvasive tools to measure liver fibrosis has permitted the screening of large patient populations. Cross-sectional study of all consecutive HIV outpatients who underwent examination by transient elastometry (FibroScan) at one HIV reference clinic during 2007. Advanced liver fibrosis (ALF) was defined as hepatic stiffness >9.5 kilopascals, which corresponds to Metavir stages F3-F4 in the liver biopsy. A total of 681 consecutive HIV-infected patients (64% injecting drug users; mean age 43; 78% male; 98% on antiretroviral therapy) had at least one valid FibroScan evaluation. ALF was diagnosed in 215 (32%) of them. In the univariate analysis, ALF was significantly associated with older age, low CD4 counts, chronic hepatitis C, past alcohol abuse, elevated ALT, high triglycerides, low cholesterol, high homeostasis model assessment (HOMA) index and exposure to didanosine and/or stavudine. In a multivariate model (OR, 95% CI), chronic hepatitis C (2.83, 1.57-5.08), past alcohol abuse (2.26, 1.37-3.74), exposure to didanosine and/or stavudine (1.85, 1.14-3.01), high HOMA index (1.25, 1.04-1.51), older age (1.09, 1.05-1.14) and elevated ALT (1.04, 1.03-1.06) remained as independently associated with ALF. Therefore, in addition to chronic hepatitis C and alcohol abuse, insulin resistance and/or exposure to dideoxy-nucleosides may contribute to ALF in HIV-infected patients.


A biocompatible and cathepsin B sensitive nanoscale system of dendritic polyHPMA-gemcitabine prodrug enhances antitumor activity markedly.
In an attempt to improve the therapeutic indices of gemcitabine (GEM), a prodrug was designed by conjugating GEM with a stimuli-responsive dendritic polyHPMA copolymer (dendritic polyHPMA-GEM) and synthesized using the one-pot method of RAFT polymerization. The prodrug with dendritic architectures was able to aggregate and form stable nanoscale systems in the order of 46 nm. The high molecular weight (HMW, 168 kDa) dendritic prodrug could biodegrade into segments of low molecular weight (LMW, 29 kDa) for excretion. The prodrug demonstrates enzyme-responsive drug release features; over 95% GEM was released from the carrier in the presence of cathepsin B within 3 h. Investigation of the cellular mechanism underlying the dendritic prodrug suggests that cytotoxicity is associated with cellular uptake and cell apoptosis. The prodrug shows good hemocompatibility and in vivo biosafety. In particular, the dendritic polymer prodrug displays high accumulation within tumors and markedly improved in vivo antitumor activity in the 4T1 murine breast cancer model compared to free GEM. The in vivo antitumor activities are characterized by a marked suppression in tumor volumes indicating much higher tumor growth inhibition (TGI, 83%) than that in GEM treatment (TGI, 36%). In addition, some tumors were eliminated. The tumor xenograft immunohistochemistry study clearly indicates that tumor apoptosis occurs through antiangiogenic effects. These results suggest that the stimuli-responsive dendritic polymer-gemcitabine has great potential as an efficient anticancer agent.


Patient-bounded extrapolation using low-dose priors for volume-of-interest imaging in C-arm CT.
Three-dimensional (3D) volume-of-interest (VOI) imaging with C-arm systems provides anatomical information in a predefined 3D target region at a considerably low x-ray dose. However, VOI imaging involves laterally truncated projections from which conventional reconstruction algorithms generally yield images with severe truncation artifacts. Heuristic based extrapolation methods, e.g., water cylinder extrapolation, typically rely on techniques that complete the truncated data by means of a continuity assumption and thus appear to be ad-hoc. It is our goal to improve the image quality of VOI imaging by exploiting existing patient-specific prior information in the workflow. A necessary initial step prior to a 3D acquisition is to isocenter the patient with respect to the target to be scanned. To this end, low-dose fluoroscopic x-ray acquisitions are usually applied from anterior-posterior (AP) and medio-lateral (ML) views. Based on this, the patient is isocentered by repositioning the table. In this work, we present a patient-bounded extrapolation method that makes use of these noncollimated fluoroscopic images to improve image quality in 3D VOI reconstruction. The algorithm first extracts the 2D patient contours from the noncollimated AP and ML fluoroscopic images. These 2D contours are then combined to estimate a volumetric model of the patient. Forward-projecting the shape of the model at the eventually acquired C-arm rotation views gives the patient boundary information in the projection domain. In this manner, we are in the position to substantially improve image quality by enforcing the extrapolated line profiles to end at the known patient boundaries, derived from the 3D shape model estimate. The proposed method was evaluated on eight clinical datasets with different degrees of truncation. The proposed algorithm achieved a relative root mean square error (rRMSE) of about 1.0% with respect to the reference reconstruction on nontruncated data, even in the presence of severe truncation, compared to a rRMSE of 8.0% when applying a state-of-the-art heuristic extrapolation technique. The method we proposed in this paper leads to a major improvement in image quality for 3D C-arm based VOI imaging. It involves no additional radiation when using fluoroscopic images that are acquired during the patient isocentering process. The model estimation can be readily integrated into the existing interventional workflow without additional hardware.


Precise measurement of the W-boson mass with the CDF II detector.
We have measured the W-boson mass M(W) using data corresponding to 2.2 fb(-1) of integrated luminosity collected in pp collisions at sqrt[s] = 1.96 TeV with the CDF II detector at the Fermilab Tevatron collider. Samples consisting of 470,126 W → eν candidates and 624,708 W → μν candidates yield the measurement M(W) = 80,387 ± 12(stat.) ± 15(syst.) = 80,387 ± 19 MeV/c2. This is the most precise measurement of the W-boson mass to date and significantly exceeds the precision of all previous measurements combined.


Developmental disorders of the fetus in some arthropod-borne virus infections.
A number of arboviruses have been associated with congenital defects in domestic aminals and man. In this review comparison is made of the temporal association between epidemics of arboviruses affecting man and animals in which there is an obvious relationship between the infection and the fetal defects, and arboviruses which cause no overt clinical symptoms in the vertebrate host but result in deformities of the fetus. The danger to the fetus following the use of live attenuated virus vaccines against several important arbovirus diseases is also examined. It is concluded that arboviruses which are capable of infecting humans or animals without producing overt clinical signs, and attenuated vaccine viruses pose the greatest threat to the fetus.


Soft-tissue reconstruction after total en bloc sacrectomy.
OBJECT Total en bloc sacrectomy is a dramatic procedure that results in extensive sacral defects. The authors present a series of patients who underwent flap reconstruction after total sacrectomy, report clinical outcomes, and provide a treatment algorithm to guide surgical care of this unique patient population. METHODS After institutional review board approval, data were collected for all patients who underwent total sacrectomy between 2002 and 2012 at The Johns Hopkins Hospital. Variables included demographic data, medical history, tumor characteristics, surgical details, postoperative complications, and clinical outcomes. All subtotal sacrectomies were excluded. RESULTS Between 2002 and 2012, 9 patients underwent total sacrectomy with flap reconstruction. Diagnoses included chordoma (n = 5), osteoblastoma (n = 1), sarcoma (n = 2), and metastatic colon cancer (n = 1). Six patients received gluteus maximus (GM) flaps with a prosthetic rectal sling following a single-stage, posterior sacrectomy. Four required additional paraspinous muscle (PSM) or pedicled latissimus dorsi (LD) fasciocutaneous flaps. Three patients underwent multistage sacrectomy with an anterior-posterior approach, 2 of whom received pedicled vertical rectus abdominis myocutaneous (VRAM) flaps, and 1 of whom received local GM, LD, and PSM flaps. Flap complications included dehiscence (n = 4) and infection (n = 1). During the 1st year of follow-up, 2 of 9 patients (22%) were able to ambulate with an assistive device by the 1st postoperative month, and 6 of 9 (67%) were ambulatory with a walker by the 3rd postoperative month. By postoperative Month 12, 5 of 9 patients (56%)-or 5 of 5 patients not lost to follow-up (100%)-were able to able to ambulate independently. CONCLUSIONS The authors' experience suggests that the GM and pedicled VRAM flaps are reliable options for softtissue reconstruction of total sacrectomy defects. For posterior-only operations, GM flaps with or without a prosthetic rectal sling are generally used. For multistage operations including a laparotomy, the authors consider the pedicled VRAM flap to be the gold standard for simultaneous reconstruction of the pelvic diaphragm and obliteration of dead space.


Functional and phenotypic differences of pure populations of stem cell-derived astrocytes and neuronal precursor cells.
Availability of homogeneous astrocyte populations would facilitate research concerning cell plasticity (metabolic and transcriptional adaptations; innate immune responses) and cell cycle reactivation. Current protocols to prepare astrocyte cultures differ in their final content of immature precursor cells, preactivated cells or entirely different cell types. A new method taking care of all these issues would improve research on astrocyte functions. We found here that the exposure of a defined population of pluripotent stem cell-derived neural stem cells (NSC) to BMP4 results in pure, nonproliferating astrocyte cultures within 24-48 h. These murine astrocytes generated from embryonic stem cells (mAGES) expressed the positive markers GFAP, aquaporin 4 and GLT-1, supported neuronal function, and acquired innate immune functions such as the response to tumor necrosis factor and interleukin 1. The protocol was applicable to several normal or disease-prone pluripotent cell lines, and the corresponding mAGES all exited the cell cycle and lost most of their nestin expression, in contrast to astrocytes generated by serum-addition or obtained as primary cultures. Comparative gene expression analysis of mAGES and NSC allowed quantification of differences between the two cell types and a definition of an improved marker set to define astrocytes. Inclusion of several published data sets in this transcriptome comparison revealed the similarity of mAGES with cortical astrocytes in vivo. Metabolic analysis of homogeneous NSC and astrocyte populations revealed distinct neurochemical features: both cell types synthesized glutamine and citrate, but only mature astrocytes released these metabolites. Thus, the homogeneous cultures allowed an improved definition of NSC and astrocyte features.


Palliative interventions for controlling vaginal bleeding in advanced cervical cancer.
Cervical cancer is the second most common cancer among women worldwide, with around 500,000 new diagnoses and 273,000 deaths per year. However, incidence and stage at diagnosis vary greatly between geographic areas and are largely dependent on the availability of a robust population screening programme. For example, in Nigeria, advanced-stage disease at presentation is common (86% to 89.3% of new cases), whereas in the UK, only 21.9% of women present with International Federation of Gynaecology and Obstetrics (FIGO) stage II+ disease. Women with advanced cancer of the cervix often need palliation for distressing symptoms, such as vaginal bleeding. Vaginal bleeding can be life threatening in advanced disease, with an incidence ranging from 0.7% to 100%. Bleeding is the immediate cause of death in 6% of women with cervical cancer and its management often poses a challenge.Thus, vaginal bleeding remains a common consequence of advanced cervical cancer. Currently, there is no systematic review that addresses palliative interventions for controlling vaginal bleeding caused by advanced cervical cancer. A systematic evaluation of the available palliative interventions is needed, to inform decision-making. To evaluate the efficacy and safety of tranexamic acid, vaginal packing (with or without formalin-soaked packs), interventional radiology or other interventions compared with radiotherapy for palliative treatment of vaginal bleeding in women with advanced cervical cancer. We searched the Cochrane Central Register of Controlled Trials (CENTRAL), Issue 2, 2015; the Cochrane Gynaecological Cancer Group Trials Register; MEDLINE from 1980 to March week 3, 2015 and EMBASE from 1980 to February week 12, 2015. We also searched registers of clinical trials, abstracts of scientific meetings and reference lists of review articles and contacted experts in the field. We handsearched citation lists of relevant studies. We searched for randomised and non-randomised comparative studies that evaluated the efficacy and safety of tranexamic acid, vaginal packing (with or without formalin-soaked packs), interventional radiology or other interventions compared with radiotherapy techniques for palliative treatment of vaginal bleeding in women with advanced cervical cancer (with or without metastasis), irrespective of publication status, year of publication or language in the review. Two review authors independently assessed whether potentially relevant studies met the inclusion criteria. We found no studies were identified for inclusion and, therefore, we analysed no data. The search strategy identified 1335 unique references of which 1160 were excluded on the basis of title and abstract. We retrieved the remaining 22 articles in full, but none satisfied the inclusion criteria. We identified only observational data from single-arm studies of women treated with formalin-soaked packs, interventional radiology or radiotherapy techniques for palliative control of vaginal bleeding in women with cervical cancer. There is no evidence from controlled trials to support or refute the use of any of the proposed interventions compared with radiotherapy. Therefore, the choice of intervention will be based on local resources. Radiotherapy techniques for managing vaginal bleeding are not readily available in resource-poor settings, where advanced cases of cervical cancer are predominant. Thus, this systematic review identified the need for a randomised controlled trial assessing the benefits and risks of palliative treatments for vaginal bleeding in women with advanced cervical cancer.


Protein intakes and serum albumin levels in a Japanese general population: NIPPON DATA90.
It is well-known that albumin is synthesized in the liver; serum albumin is a major component of serum proteins. However, it has not been well elucidated how dietary protein intakes are associated with serum albumin levels in general populations without extreme malnutrition. We cross-sectionally investigated in the representative Japanese the association between dietary protein intake and serum albumin levels. A total of 7715 subjects (3220 men and 4495 women, aged 30 years or more) with measurement of serum albumin who participated in both the National Survey on Circulatory Disorders in 1990 and the National Nutrition Survey in 1990 were analyzed in the present analysis. Multiple-adjustments were performed with linear regression models to estimate the association between serum albumin levels and animal or vegetable protein intake adjusting for age and body mass index. The very weak positive association between animal protein and serum albumin levels was observed. On the other hand, there was no clear association observed between vegetable protein and serum albumin levels. Regardless of sex and models, age was inversely associated with serum albumin levels with statistically significance, and standardized coefficients of age were considerably larger in both sexes than other variables. Adjustment for body mass index hardly altered the coefficients of animal or vegetable protein intake, but adjustment for total cholesterol clearly attenuated the relationship between animal protein intake and serum albumin levels. Present analysis indicated the possibility that animal protein intake was related with serum albumin levels, while vegetable protein intake was not related.


Development and validation of a qPCR-based method for quantifying Shiga toxin-encoding and other lambdoid bacteriophages.
To address whether seasonal variability exists among Shiga toxin-encoding bacteriophage (Stx phage) numbers on a cattle farm, conventional plaque assay was performed on water samples collected over a 17 month period. Distinct seasonal variation in bacteriophage numbers was evident, peaking between June and August. Removal of cattle from the pasture precipitated a reduction in bacteriophage numbers, and during the winter months, no bacteriophage infecting Escherichia coli were detected, a surprising occurrence considering that 10(31) tailed-bacteriophages are estimated to populate the globe. To address this discrepancy a culture-independent method based on quantitative PCR was developed. Primers targeting the Q gene and stx genes were designed that accurately and discriminately quantified artificial mixed lambdoid bacteriophage populations. Application of these primer sets to water samples possessing no detectable phages by plaque assay, demonstrated that the number of lambdoid bacteriophage ranged from 4.7 x 10(4) to 6.5 x 10(6) ml(-1), with one in 10(3) free lambdoid bacteriophages carrying a Shiga toxin operon (stx). Specific molecular biological tools and discriminatory gene targets have enabled virus populations in the natural environment to be enumerated and similar strategies could replace existing propagation-dependent techniques, which grossly underestimate the abundance of viral entities.


Socioeconomic differences in the use of ill-defined causes of death in 16 European countries.
Cause-of-death data linked to information on socioeconomic position form one of the most important sources of information about health inequalities in many countries. The proportion of deaths from ill-defined conditions is one of the indicators of the quality of cause-of-death data. We investigated educational differences in the use of ill-defined causes of death in official mortality statistics. Using age-standardized mortality rates from 16 European countries, we calculated the proportion of all deaths in each educational group that were classified as due to "Symptoms, signs and ill-defined conditions". We tested if this proportion differed across educational groups using Chi-square tests. The proportion of ill-defined causes of death was lower than 6.5% among men and 4.5% among women in all European countries, without any clear geographical pattern. This proportion statistically significantly differed by educational groups in several countries with in most cases a higher proportion among less than secondary educated people compared with tertiary educated people. We found evidence for educational differences in the distribution of ill-defined causes of death. However, the differences between educational groups were small suggesting that socioeconomic inequalities in cause-specific mortality in Europe are not likely to be biased.