Attention problems, inhibitory control, and intelligence index overlapping genetic factors: a study in 9-, 12-, and 18-year-old twins.
It is assumed that attention problems (AP) are related to impaired executive functioning. We investigated the association between AP and inhibitory control and tested to what extent the association was due to genetic factors shared with IQ. Data were available from 3 independent samples of 9-, 12-, and 18-year-old twins and their siblings (1,209 participants). AP were assessed with checklists completed by multiple informants. Inhibitory control was measured with the Stroop Color Word Task (Stroop, 1935), and IQ with the Wechsler Intelligence Scale for Children (Wechsler et al., 2002) or Wechsler Adult Intelligence Scale (Wechsler, 1997). AP and inhibitory control were only correlated in the 12-year-old cohort (r = .18), but appeared non-significant after controlling for IQ. Significant correlations existed between AP and IQ in 9- and 12-year olds (r = -.26/-.34). Inhibitory control and IQ were correlated in all cohorts (r = -.16, -.24 and -.35, respectively). Genetic factors that influenced IQ also influenced inhibitory control. We conclude that the association between AP and inhibitory control as reported in the literature may largely derive from genetic factors that are shared with IQ.


An S-adenosyl Methionine Synthetase (SAMS) Gene from Andropogon virginicus L. Confers Aluminum Stress Tolerance and Facilitates Epigenetic Gene Regulation in Arabidopsis thaliana.
Candidate clones which conferred Al tolerance to yeast transformants (TFs) were obtained from a cDNA library derived from a highly Al-tolerant poaceae, Andropogon virginicus L. One such clone, AL3A-4, encoded an S-adenosyl methionine synthetase (SAMS) gene. A full-length cDNA was obtained by 5'-RACE, designated AvSAMS1, and introduced into Arabidopsis thaliana to investigate its biological functions under Al stress. Two TF plant lines both showed higher tolerance than the Col-0 ecotype (non-TF) not only for Al stress, but also for Cu, Pb, Zn and diamide stresses, suggesting the AvSAMS1 was a multiple tolerance gene. More than 40 of A. thaliana Al response-genes (Al induced genes and Al repressed genes) were selected from microarray results and then used for investigations of DNA or histone methylation status under Al stress in Col-0 and the AvSAMS1 TF line. The results indicated that Al stress caused alterations of methylation status in both DNA and histone H3 (H3K4me3 and H3K9me3) and that these alterations were different between the AvSAMS1 TF and Col-0, suggesting the differences were AvSAMS1-gene dependent. These results suggested the existence of AvSAMS1-related epigenetic gene-regulation under Al stress.


Trafficking of Endogenous Immunoglobulins by Endothelial Cells at the Blood-Brain Barrier.
The Blood-Brain Barrier (BBB) restricts access of large molecules to the brain. The low endocytic activity of brain endothelial cells (BECs) is believed to limit delivery of immunoglobulins (IgG) to the brain parenchyma. Here, we report that endogenous mouse IgG are localized within intracellular vesicles at steady state in BECs in vivo. Using high-resolution quantitative microscopy, we found a fraction of endocytosed IgG in lysosomes. We observed that loss of pericytes (key components of the BBB) in pdgf-b(ret/ret) mice affects the intracellular distribution of endogenous mouse IgG in BECs. In these mice, endogenous IgG was not detected within lysosomes but instead accumulate at the basement membrane and brain parenchyma. Such IgG accumulation could be due to reduced lysosomal clearance and increased sorting to the abluminal membrane of BECs. Our results suggest that, in addition to low uptake from circulation, IgG lysosomal degradation may be a downstream mechanism by which BECs further restrict IgG access to the brain.


Estimating iron overload in patients with suspected liver disease and elevated serum ferritin.
Iron status evaluation in patients with suspected liver disease and elevated serum ferritin is often challenging because hyperferritinemia does not always indicate iron overload. A reliable approach to estimate iron overload without exposing the patient to unnecessary investigations would help the clinician to identify patients who may take advantage of iron-removal therapy. We analyzed all liver biopsies, including measurement of hepatic iron concentration, performed at the University Hospital Zurich from 1997 to 2010 to identify clinical and laboratory predictors of iron overload in patients with elevated serum ferritin (n = 147). Hyperferritinemia was predictive of iron overload only in patients with a high level of serum ferritin (>2000 μg/L). In patients with moderate hyperferritinemia, liver transaminases inversely correlated with hepatic iron concentration. A combination of both parameters expressed as ferritin/aspartate transaminase ratio was highly predictive of tissue iron overload (sensitivity 83.3%, specificity 78.6%). Receiver operating characteristic analysis resulted in an area under the curve of 0.83. We established a simple and reliable method to correctly estimate iron overload in patients with suspected liver disease and elevated serum ferritin.


AMPK signaling in neuronal polarization: Putting the brakes on axonal traffic of PI3-Kinase.
Neuronal polarization, the process by which neurons form multiple dendrites and an axon from the soma, is the first critical step in the formation and function of neural networks. Polarization begins with the rapid extension of a single neurite to produce an axon of impressive size and complex geometry, while the remaining sister neurites differentiate into dendrites. The extensive biosynthesis required to produce an axon therefore necessitates coordination with cellular energy status to ensure an ample energy supply. Our recent work shows that activity of the AMP-activated protein kinase (AMPK), the bio-energy sensor responsible for maintaining cellular energy homeostasis in all eukaryotic cells, plays an important role in the initiation of axonal growth. AMPK phosphorylates the cargo-binding light chain of the Kif5 motor protein, leading to dissociation of the phosphatidylinositol 3-Kinase (PI3K) from the motor complex. The mislocation of PI3K, which is normally enriched at the axonal tip for extension and differentiation, results in a lack of neurite specification and neuron polarization. These findings reveal a link between cellular bioenergy homeostasis and neuron morphogenesis, and suggest a novel cellular mechanism underlying the long-term neurological abnormalities as a consequence of bioenergy deficiency during early brain development.


Acute effects of deep breathing for a short duration (2-10 minutes) on pulmonary functions in healthy young volunteers.
Breathing is the most vital function for maintenance of life. Slow and deep breathing is an integral part of Pranayama and it reduces dead space ventilation and renews air throughout the lungs. The reported beneficial effects of deep breathing as a part of either long term or short term practice of Pranayama are well documented. However our knowledge about the effects of a few minutes' of deep breathing on human ventilatory parameters is poor. In the present study, we examined the relationship between exposure to short duration of deep breathing and performance on pulmonary function tests before and after the deep breathing. The study was conducted in a homogenous group of 12 volunteers containing 4 females and 8 males who were well trained in pulmonary function testing (PFT) before the start of the study. The volunteers performed deep breathing (DB) exercise for 2, 5 and 10 minutes at the rate of 6 breaths per minute under guidance, and the duration of DB exercise for that day was randomly selected for each group. PFT was done before and after the DB exercise. There was a significant (P < 0.05) increase in vital capacity (VC) after 2 and 5 minutes' DB exercise and a consistent improvement in tidal volume (TV) and minute ventilation (MV) after the DB exercise in all the three groups, though it wasn't statistically significant. There was a significant (P < 0.05) increase in forced vital capacity (FVC) after 2 minutes' of DB exercise and a consistent increase in all the three groups in forced inspiratory vital capacity (FIVC) and peak inspiratory flow rate (PIFR), though this increase was not statistically significant. This shows that deep breathing exercise, even for a few minutes' duration is beneficial for the lung functions.


Ghrelin in patients with irritable bowel syndrome.
General gastrointestinal dysmotility occurs in patients with irritable bowel syndrome (IBS). Ghrelin seems to play an important role in regulating gastrointestinal motility. The present study was undertaken, therefore, to establish the possible role of ghrelin in the pathophysiology of IBS. Thirty-seven patients with IBS (19 had IBS-constipation and 18 IBS-diarrhoea) were included in this study. Ten healthy volunteers served as controls. After overnight fast, blood samples were drawn from patients and controls, and a gastroduodenal endoscopy was performed. Biopsies were taken from oxyntic mucosa and duodenum. Ghrelin cell density was determined by computer image analysis after immunohistochemical staining of the tissues. Total and active ghrelin were detected in tissue extracts and plasma by commercially available RIA and ELISA Kits. The density of ghrelin-immunoreactive cells in the oxyntic mucosa was significantly lower in IBS-constipation and significantly higher in IBS-diarrhoea patients than healthy controls (P<0.0001 and <0.0001, respectively). There was no statistical difference in total or active ghrelin between IBS patients and controls, regarding tissue extracts or plasma. In order to compensate for the increase and decrease in the ghrelin cell density, the synthesis and release of ghrelin may be decreased and increased in IBS-diarrhoea and IBS-constipation patients, respectively. It has been speculated that this compensatory mechanism may be subjected from time to time to fatigue with the subsequent increased and decreased synthesis and release of ghrelin in IBS-diarrhoea and IBS-constipation with a subsequent intermittent diarrhoea or constipation seen in these patients, respectively.


Dendritic cell-specific disruption of TGF-β receptor II leads to altered regulatory T cell phenotype and spontaneous multiorgan autoimmunity.
In vitro data and transgenic mouse models suggest a role for TGF-β signaling in dendritic cells (DCs) to prevent autoimmunity primarily through maintenance of DCs in their immature and tolerogenic state characterized by low expression of MHC class II (MHCII) and costimulatory molecules and increased expression of IDO, among others. To test whether a complete lack of TGF-β signaling in DCs predisposes mice to spontaneous autoimmunity and to verify the mechanisms implicated previously in vitro, we generated conditional knockout (KO) mice with Cre-mediated DC-specific deletion of Tgfbr2 (DC-Tgfbr2 KO). DC-Tgfbr2 KO mice die before 15 wk of age with multiorgan autoimmune inflammation and spontaneous activation of T and B cells. Interestingly, there were no significant differences in the expression of MHCII, costimulatory molecules, or IDO in secondary lymphoid organ DCs, although Tgfbr2-deficient DCs were more proinflammatory in vitro and in vivo. DC-Tgfbr2 KO showed attenuated Foxp3 expression in regulatory T cells (Tregs) and abnormal expansion of CD25(-)Foxp3(+) Tregs in vivo. Tgfbr2-deficient DCs secreted elevated levels of IFN-γ and were not capable of directing Ag-specific Treg conversion unless in the presence of anti-IFN-γ blocking Ab. Adoptive transfer of induced Tregs into DC-Tgfbr2 KO mice partially rescued the phenotype. Therefore, in vivo, TGF-β signaling in DCs is critical in the control of autoimmunity through both Treg-dependent and -independent mechanisms, but it does not affect MHCII and costimulatory molecule expression.


Outbreak of Saprochaete clavata Sepsis in Hematology Patients: Combined Use of MALDI-TOF and Sequencing Strategy to Identify and Correlate the Episodes.
New fungal species are increasingly reported in immunocompromised patients. Saprochaete clavata (S. clavata), an ascomycetous fungus formerly called Geotrichum clavatum, is intrinsically resistant to echinocandins and is often misidentified. We describe a cluster of seven S. clavata infections in hospitalized hematology patients who developed this rare fungemia within a span of 11 months. Three of the seven patients died. Identification of the isolates was determined only with the Saramis database of VitekMS system and sequencing of the internal transcribed spacer (ITS) region. Clonal relatedness of the isolates was determined by matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF) analysis; clonal correlation between the strains was investigated by means of phylogenetic analysis, based on single-nucleotide variants (SNPs). Clinical presentation, 1-3 β-D-glucan (BG) and galactomannan (GM) antigen results and analysis of possible sources of contamination are also described with a prospective case-control study of the outbreak. MALDI-TOF MS-Vitek (bioMerieux, Marcy l'Etoile, France) failed to identify the six isolates, while SARAMIS (bioMerieux, Marcy l'Etoile, France) identified the isolates as S. clavata. Initially, Vitek 2 identified the strains as Geotrichum capitatum in two of the seven cases. Molecular identification gave 99% homology with S. clavata. BG was positive in three out of six patients (range 159 to >523 pg/ml), GM results were always negative. All the isolates were resistant to echinocandins (anidulafungin, micafungin, and caspofungin) and Fluconazole, but susceptible to Flucytosine and Voriconazole. One isolate showed acquired resistance to Flucytosine and Amphotericin B during treatment. Both the correlation-based dendrograms obtained by MALDI-TOF MS (Bruker Daltonics) and MS-Vitek not only clustered six of the seven bloodstream infection (BSI) isolates in the same group, but also showed their strong relatedness. Phylogenetic analysis using SNPrelate showed that the seven samples recorded during the investigation period clustered together. We observed a split between one case and the remainder with a node supported by a z-score of 2.3 (p-value = 0.021) and 16 mutations unique to each branch. The use of proteomics for identification and evaluation of strain clonality in outbreaks of rare pathogens is a promising alternative to laborious and time-consuming molecular methods, even if molecular whole-genome sequencing (WGS) typing will still remain the reference method for rare emergent pathogens.


Odorant Inhibition in Mosquito Olfaction.
How chemical signals are integrated at the peripheral sensory system of insects is still an enigma. Here we show that when coexpressed with Orco in Xenopus oocytes, an odorant receptor from the southern house mosquito, CquiOR32, generated inward (regular) currents when challenged with cyclohexanone and methyl salicylate, whereas eucalyptol and fenchone elicited inhibitory (upward) currents. Responses of CquiOR32-CquiOrco-expressing oocytes to odorants were reduced in a dose-dependent fashion by coapplication of inhibitors. This intrareceptor inhibition was also manifested in vivo in fruit flies expressing the mosquito receptor CquiOR32, as well in neurons on the antennae of the southern house mosquito. Likewise, an orthologue from the yellow fever mosquito, AaegOR71, showed intrareceptor inhibition in the Xenopus oocyte recording system and corresponding inhibition in antennal neurons. Inhibition was also manifested in mosquito behavior. Blood-seeking females were repelled by methyl salicylate, but repellence was significantly reduced when methyl salicylate was coapplied with eucalyptol.


Contribution of hand motor circuits to counting.
The finding that number processing activates a cortical network partly overlapping that recruited for hand movements has renewed interest in the relationship between number and finger representations. Further evidence about a possible link between fingers and numbers comes from developmental studies showing that finger movements play a crucial role in learning counting. However, increased activity in hand motor circuits during counting may unveil unspecific processes, such as shifting attention, reciting number names, or matching items with a number name. To address this issue, we used transcranial magnetic stimulation to measure changes in corticospinal (CS) excitability during a counting task performed silently and using either numbers or letters of the alphabet to enumerate items. We found an increased CS excitability of hand muscles during the counting task, irrespective of the use of numbers or letters, whereas it was unchanged in arm and foot muscles. Control tasks allowed us to rule out a possible influence of attention allocation or covert speech on CS excitability increase of hand muscles during counting. The present results support a specific involvement of hand motor circuits in counting because no CS changes were found in arm and foot muscles during the same task. However, the contribution of hand motor areas is not exclusively related to number processing because an increase in CS excitability was also found when letters were used to enumerate items. This finding suggests that hand motor circuits are involved whenever items have to be put in correspondence with the elements of any ordered series.


Epidemiology of primary CNS lymphoma.
In the beginning of the nineties the National Cancer Institute Surveillance, Epidemiology, and End Results Program calculated the incidence of primary central nervous system non-Hodgkin's lymphoma (PCNSL) as 1:100000. The incidence of PCNSL has been increasing since the seventies in immunocompetent patients. The main increase, however, is taking place since the mid-eighties and is due to the increase of immunodeficiency and immunosuppression. The risk is 2-6% in AIDS patients according to clinical data and will probably further increase with the length of survival in these patients. Transplant patients carry a risk of 1-5% to develop a PCNSL. The risk is 1-2% for renal, and 2-7% for cardiac, lung or liver transplant recipients. Patients with congenital immune deficiency have a risk of 4%. PCNSL may also present as a secondary malignancy.


Comparison of [11C]flumazenil and [18F]FDG as PET markers of epileptic foci.
Recent PET results indicate that the benzodiazepine (BZ) receptor density measured with the BZ receptor antagonist [11C]flumazenil is reduced in human epileptic foci. The present study examines the applicability of this finding in the presurgical investigation of patients with intractable partial epilepsy. In eight patients, the PET measurements were performed after injection of the BZ receptor antagonist [11C]flumazenil and [2-18F]2-deoxy-2-fluoro-D-glucose ([18F]FDG)--a tracer for measurements of the rate of regional glucose metabolism. The focus localising ability of the two PET tracers was examined using extra--and intracranial EEG recordings as reference. The focus was first determined visually on the PET images obtained after a bolus injection of each of the PET tracers. Its anatomical localisation and spatial delimitation was then evaluated for each patient with a computerised anatomical brain atlas. [11C]flumazenil was found to be a more sensitive and accurate focus localiser than [18F]FDG. This observation was valid both for quantified and non-quantified images. In the preoperative diagnosis of epileptic foci, the PET measurements of BZ receptors may be a suitable and, in some cases, superior method to the generally used "[18F]FDG-PET" method.


Pulsed electric field treatment for bacteria reduction and its impact on hospital wastewater.
During the last years the pulsed electric field (PEF) method entered several fields of application. A promising application is the decontamination of hospital wastewater effluents, which are loaded with pathogenic and increasingly with antibiotic-resistant bacteria. For this study, Pseudomonas putida suspended in buffer solution or wastewater from university hospital was used as reference strain. To prove whether the descendent of the survival bacteria develop an adaptation to electric field, surviving PEF treated bacteria were recultivated and pulsed in serial experiments with 10 pulses (100kVcm(-1) and 600ns pulse duration). This procedure was repeated for 30 generations. The inactivation rate was calculated with 3.5+/-0.8 log of colony forming units and remained constant over 30 cycles. Investigations of the variable intergenic spacer region of the ribosomal operon demonstrated no visible changes in this highly variable part of the genome structure during the serial PEF treatment experiments. The mutagenicity of PEF treated hospital wastewater, buffer solutions and tap water was analyzed by the umu-test. Most hospital wastewater samples exhibit a considerable genotoxicity already before PEF treatment, but this was not increased by the PEF treatment, not even for higher treatments energies over 250JmL(-1). No genotoxicity was induced in buffer solutions and tap water by PEF treatment. This study supports, that PEF treatment is a sustainable non-chemical method for bacterial decontamination without any adverse effects.


Association between neighborhood need and spatial access to food stores and fast food restaurants in neighborhoods of colonias.
To determine the extent to which neighborhood needs (socioeconomic deprivation and vehicle availability) are associated with two criteria of food environment access: 1) distance to the nearest food store and fast food restaurant and 2) coverage (number) of food stores and fast food restaurants within a specified network distance of neighborhood areas of colonias, using ground-truthed methods. Data included locational points for 315 food stores and 204 fast food restaurants, and neighborhood characteristics from the 2000 U.S. Census for the 197 census block group (CBG) study area. Neighborhood deprivation and vehicle availability were calculated for each CBG. Minimum distance was determined by calculating network distance from the population-weighted center of each CBG to the nearest supercenter, supermarket, grocery, convenience store, dollar store, mass merchandiser, and fast food restaurant. Coverage was determined by calculating the number of each type of food store and fast food restaurant within a network distance of 1, 3, and 5 miles of each population-weighted CBG center. Neighborhood need and access were examined using Spearman ranked correlations, spatial autocorrelation, and multivariate regression models that adjusted for population density. Overall, neighborhoods had best access to convenience stores, fast food restaurants, and dollar stores. After adjusting for population density, residents in neighborhoods with increased deprivation had to travel a significantly greater distance to the nearest supercenter or supermarket, grocery store, mass merchandiser, dollar store, and pharmacy for food items. The results were quite different for association of need with the number of stores within 1 mile. Deprivation was only associated with fast food restaurants; greater deprivation was associated with fewer fast food restaurants within 1 mile. CBG with greater lack of vehicle availability had slightly better access to more supercenters or supermarkets, grocery stores, or fast food restaurants. Increasing deprivation was associated with decreasing numbers of grocery stores, mass merchandisers, dollar stores, and fast food restaurants within 3 miles. It is important to understand not only the distance that people must travel to the nearest store to make a purchase, but also how many shopping opportunities they have in order to compare price, quality, and selection. Future research should examine how spatial access to the food environment influences the utilization of food stores and fast food restaurants, and the strategies used by low-income families to obtain food for the household.


Cephalometry in adults and children with neurofibromatosis type 1: Implications for the pathogenesis of sphenoid wing dysplasia and the "NF1 facies".
Neurofibromatosis type 1 (NF1) is a common, autosomal dominant tumor-predisposition disorder that arises secondary to mutations in the tumor suppressor gene NF1. Cephalometry is an inexpensive, readily available and non-invasive technique that is under-utilized in studying the NF1 craniofacial phenotype. An analysis of NF1 cephalometry was first published by Heervä et al. in 2011. We expand here on that first investigation with a larger cohort of adult and pediatric patients affected with NF1 and sought objective insight into the NF1 facies, said to feature hypertelorism and a broad nasal base, from cephalometric analysis. We obtained cephalograms from 101 patients with NF1 (78 adults and 23 children) from two NF1 protocols at the National Institutes of Health. Each subject had an age-, gender- and ethnicity-matched control. We used Dolphin software to make the cephalometric measurements. We assessed the normality of differences between paired samples using the Shapiro-Wilk test and evaluated the significance of mean differences using paired t-tests and adjusted for multiple testing. We explored the relationship between the cephalometric measurements and height, head circumference and interpupillary distance. In this dataset of American whites with NF1, we confirmed in a modestly larger sample many of the findings found by Heerva et al. in an NF1 Finnish cohort. We found a shorter maxilla, mandible, cranial base, (especially anteriorly, p = 0.0001) and diminished facial height in adults, but not children, with NF1. Only one adult exhibited hypertelorism. The cephalometric differences in adults arise in part from cranial base shortening and thus result in a shorter face, mid-face hypoplasia, reduced facial projection, smaller jaw, and increased braincase globularity. In addition, we suggest that NF1 sphenoid bone shortening, a common event, is consistent with an intrinsic NF1 bone cell defect, which renders the bone more vulnerable to a random "second hit" in NF1, leading to sphenoid wing dysplasia, a rare event.


High resolution interrogation of tilted fiber grating SPR sensors from polarization properties measurement.
The generation of surface plasmon resonances (SPRs) in gold-coated weakly tilted fiber Bragg gratings (TFBGs) strongly depends on the state of polarization of the core guided light. Recently, it was demonstrated that rotating the linear state of polarization of the guided light by 90° with respect to the grating tilt allows to turn the SPR on and off. In this work, we measure the Jones matrix associated to the TFBG transmission properties in order to be able to analyze different polarization-related parameters (i.e. dependency on wavelength of polarization dependent loss and first Stokes parameter). As they contain the information about the SPR, they can be used as a robust and accurate demodulation technique for refractometry purposes. Unlike other methods reported so far, a tight control of the input state of polarization is not required. The maximum error on refractive index measurement has been determined to be ~1 10(-5) refractive index unit (RIU), 5 times better than intensity-based measurements on the same sensors.


Metabolic flux profiling of Pichia pastoris grown on glycerol/methanol mixtures in chemostat cultures at low and high dilution rates.
The metabolic pathways associated with the tricarboxylic acid cycle intermediates of Pichia pastoris were studied using biosynthetically directed fractional 13C labelling. Cells were grown aerobically in a chemostat culture fed at two dilution rates (1.39x10(-5) s-1 and 4.44x10(-5) s-1) with varying mixtures of glycerol and methanol as sole carbon sources. The results show that, with co-assimilation of methanol, the common amino acids are synthesized as in P. pastoris cells grown on glycerol only. During growth at the lower dilution rate, when both substrates are entirely consumed, the incorporation of methanol into the biomass increases as the methanol fraction in the feed is increased. Moreover, the co-assimilation of methanol impacts on how key intermediates of the pentose phosphate pathway (PPP) are synthesized. In contrast, such an impact on the PPP is not observed at the higher dilution rate, where methanol is only partially consumed. This finding possibly indicates that the distribution of methanol carbon into assimilatory and dissimilatory (direct oxidation to CO2) pathways are different at the two dilution rates. Remarkably, distinct flux ratios were registered at each of the two growth rates, while the dependency of the flux ratios on the varying fraction of methanol in the medium was much less pronounced. This study brings new insights into the complex regulation of P. pastoris methanol metabolism in the presence of a second carbon source, revealing important implications for biotechnological applications.


Hypericum perforatum treatment: effect on behaviour and neurogenesis in a chronic stress model in mice.
Extracts of Hypericum perforatum (St. John's wort) have been traditionally recommended for a wide range of medical conditions, in particular mild-to-moderate depression. The present study was designed to investigate the effect of Hypericum perforatum treatment in a mouse model of anxiety/depressive-like behavior, induced by chronic corticosterone administration. CD1 mice were submitted to 7 weeks corticosterone administration and then behavioral tests as Open Field (OF), Novelty-Suppressed Feeding (NSF), Forced Swim Test (FST) were performed. Cell proliferation in hippocampal dentate gyrus (DG) was investigated by both 5-bromo-2'-deoxyuridine (BrdU) and doublecortin (DCX) immunohistochemistry techniques and stereological procedure was used to quantify labeled cells. Golgi-impregnation method was used to evaluate changes in dendritic spines in DG. Hypericum perforatum (30 mg/Kg) has been administered for 3 weeks and then neural development in the adult hippocampus and behavioral changes have been examined. The anxiety/depressive-like state due to chronic corticosterone treatment was reversed by exogenous administration of Hypericum perforatum; the proliferation of progenitor cells in mice hippocampus was significantly reduced under chronic corticosterone treatment, whereas a long term treatment with Hypericum perforatum prevented the corticosterone-induced decrease in hippocampal cell proliferation. Corticosterone-treated mice exhibited a reduced spine density that was ameliorated by Hypericum perforatum administration. These results provide evidence of morphological adaptations occurring in mature hippocampal neurons that might underlie resilient responses to chronic stress and contribute to the therapeutic effects of chronic Hypericum perforatum treatment.


The glycoprotein fibulin-3 regulates morphology and motility of olfactory ensheathing cells in vitro.
The primary olfactory pathway in adult mammals has retained a remarkable potential for self-repair. A specialized glial cell within the olfactory nerve, called olfactory ensheathing cell (OEC), and their associated extracellular matrix are thought to play an important role during regenerative events in this system. To gain insight into novel molecules that could mediate the OEC-supported growth of axons within the olfactory nerve, gene expression profiling experiments were conducted which revealed high expression of the glycoprotein fibulin-3 in OECs. This observation was confirmed with quantitative PCR. In vivo, the distribution of all members of the fibulin family, fibulin-3 included, was localized to the lamina propria underneath the olfactory epithelium, in close association within olfactory nerve bundles. To manipulate fibulin-3 gene expression in cultured OECs, lentiviral vector constructs were designed to either transgenically express or knock-down fibulin-3. Experimental data showed that increased levels of fibulin-3 induced profound morphological changes in cultured OECs, impeded with their migratory abilities and also suppressed OEC-mediated neurite outgrowth. Knock-down of fibulin-3 levels resulted in reduced OEC proliferation. In conclusion, the data provide novel insights into a putative role for fibulin-3 in the regulation of cell migration and neurite outgrowth within the primary olfactory pathway.


Body weight and health from early to mid-adulthood: a longitudinal analysis.
We analyze the influence of body weight in early adulthood, and changes in weight over time, on self-rated health as people age into middle adulthood. While prior research has focused on cross-sectional samples of older adults, we use longitudinal data from the NHANES I Epidemiologic Follow-up Study and double-trajectory latent growth models to study the association between body mass index (BMI) and self-rated health trajectories over 20 years. Results indicate that high BMI in early adulthood and gaining more weight over time are both associated with a faster decline in health ratings. Among white women only, those with a higher BMI at the baseline also report lower initial self-rated health. A small part of the weight-health association is due to sociodemographic factors, but not baseline health behaviors or medical conditions. The findings provide new support for the cumulative disadvantage perspective, documenting the increasing health inequalities in a cohort of young adults.


In Vivo Confocal Microscopy Demonstrates Bilateral Loss of Endothelial Cells in Unilateral Herpes Simplex Keratitis.
To report bilateral corneal endothelial cell density (ECD), as well as its correlation with subbasal nerve changes, in patients with unilateral herpes simplex keratitis (HSK). Thirty-six eyes of 36 patients with corneal scarring caused by HSK, as well as their respective contralateral clinically unaffected eyes, were prospectively studied and compared with 26 eyes of 26 healthy volunteers. In vivo confocal microscopy and corneal sensation of the central cornea were performed bilaterally in all patients and in one random eye of controls. The ECD and subbasal corneal nerve density, including the lengths of total nerves, main trunks, and branches were evaluated and correlated to central corneal sensation. The ECD was significantly lower in eyes affected with HSK than in controls (2304 ± 578 vs. 2940 ± 370 cells/mm(2), P < 0.0001). Surprisingly, lower ECD was also detected in contralateral clinically unaffected eyes (2548 ± 423), compared to controls (P = 0.02). Both affected and contralateral eyes showed decrease in total nerve length, compared to controls (10.0 ± 6.3 vs. 17.6 ± 6.3 vs. 21.9 ± 4.3 mm/mm2, respectively; P < 0.05 for all). The ECD correlated positively with total nerve length (r = 0.39, P = 0.0009) and with corneal sensation (r = 0.31, P = 0.009). In vivo confocal microscopy findings demonstrated alterations in corneal ECD in both affected and clinically unaffected contralateral eyes of patients with unilateral HSK. Moreover, the positive significant correlation between the ECD and the subbasal nerve density may suggest a potential link between corneal innervation and corneal endothelial cell homeostasis.


How Bachelorhood and Migration Increase the HIV Transmission Risk Through Commercial Sex in China?
In China, the serious involuntary bachelorhood due to sex ratio imbalance in decades is expected to dramatically increase the spread of HIV through heterosexual contact. However, the higher HIV transmission risk and its correlates among never married men in rural China are not well understood. This study explored whether and how bachelorhood and migration increased the HIV transmission risk through commercial sex. By combining two cross-sectional survey data from never married men in rural areas and male migrants (including both married and unmarried men) in urban areas, a total of 1030 participants who were never married and age 28 and above or married male migrants were included in this study. The results show that both bachelorhood and migration put the never married male migrants at particularly higher HIV transmission risk by increasing both the possibility of engaging in commercial sex, and the frequency and inconsistency of condom use in commercial sex. Selection bias into marriage and neighborhood characteristics associated with exposure to commercial sexual risk may partly explain why male migrants that never married had a higher commercial sex related risk than that of married male migrants and never married non-migrant males.


Built Environments and Cardiometabolic Morbidity and Mortality in Remote Indigenous Communities in the Northern Territory, Australia.
The health of Indigenous Australians is dramatically poorer than that of the non-Indigenous population. Amelioration of these differences has proven difficult. In part, this is attributable to a conceptualisation which approaches health disparities from the perspective of individual-level health behaviours, less so the environmental conditions that shape collective health behaviours. This ecological study investigated associations between the built environment and cardiometabolic mortality and morbidity in 123 remote Indigenous communities representing 104 Indigenous locations (ILOC) as defined by the Australian Bureau of Statistics. The presence of infrastructure and/or community buildings was used to create a cumulative exposure score (CES). Records of cardiometabolic-related deaths and health service interactions for the period 2010-2015 were sourced from government department records. A quasi-Poisson regression model was used to assess the associations between built environment "healthfulness" (CES, dichotomised) and cardiometabolic-related outcomes. Low relative to high CES was associated with greater rates of cardiometabolic-related morbidity for two of three morbidity measures (relative risk (RR) 2.41-2.54). Cardiometabolic-related mortality was markedly greater (RR 4.56, 95% confidence interval (CI), 1.74-11.93) for low-CES ILOCs. A lesser extent of "healthful" building types and infrastructure is associated with greater cardiometabolic-related morbidity and mortality in remote Indigenous locations. Attention to environments stands to improve remote Indigenous health.


Mycobacterium tuberculosis-specific and MHC class I-restricted CD8+ T-cells exhibit a stem cell precursor-like phenotype in patients with active pulmonary tuberculosis.
The nature and longevity of the T-cell response directed against Mycobacterium tuberculosis (MTB) are important for effective pathogen containment. We analyzed ex vivo the nature of MTB antigen-specific T-cell responses directed against the MTB secreted antigens Rv0288, Rv1886c, Rv3875, the antigens Rv2958c, Rv2957, and Rv0447c (intracellular, non-secreted enzymes) in blood from Korean patients with active tuberculosis (TB). MTB-specific T-cell function was defined by intracellular cytokine production (interleukin (IL)-2, interferon gamma, tumour necrosis factor alpha, and IL-17) and by multimer-guided (HLA-A*02:01 and HLA-A*24:02) analysis of epitope-specific CD8+ T-cells, along with phenotypic markers (CD45RA and CCR7), CD107a, a marker for degranulation, and CD127 co-staining for T-cell differentiation and homing. Cytokine production analysis underestimated the frequencies of MTB antigen-specific T-cells defined by major histocompatibility complex (MHC) class I-peptide multimer analysis. We showed that MTB antigen-specific CD8+ T-cells exhibit a distinct marker profile associated with the nature of the MTB antigens, i.e., Rv0288, Rv1886c, and Rv3875-reactive T-cells clustered in the precursor T-cell compartment, whereas Rv2958c, Rv2957, and Rv0447c-reactive T-cells were associated with the terminally differentiated T-cell phenotype, in the patient cohort. Rv0288, Rv1886c, and Rv3875-specific CD8+ T-cells were significantly enriched for CD107a+ T-cells in HLA-A*02:01 (p<0.0001) and HLA-A*24:02 (p=0.0018) positive individuals, as compared to Rv2958c, Rv2957, and Rv0447c antigens. CD127 (IL-7 receptor)-expressing T-cells were enriched in HLA-A*02:01-positive individuals for the Rv0288, Rv1886c, and Rv3875 specificities (p=0.03). A high proportion of antigen-specific T-cells showed a precursor-like phenotype (CD45RA+CCR7+) and expressed the stem cell-associated markers CD95 and c-kit. These data show that MTB-specific T-cells can express stem cell-like features; this is associated with the nature of the MTB antigen and the genetic background of the individual.


Human brain beta-hydroxybutyrate and lactate increase in fasting-induced ketosis.
Ketones are known to constitute an important fraction of fuel for consumption by the brain, with brain ketone content generally thought to be low. However, the recent observation of 1-mmol/L levels of brain beta-hydroxybutyrate (BHB) in children on the ketogenic diet suggests otherwise. The authors report the measurement of brain BHB and lactate in the occipital lobe of healthy adults using high field (4-T) magnetic resonance spectroscopy, measured in the nonfasted state and after 2- and 3-day fasting-induced ketosis. A 9-mL voxel located in the calcarine fissure was studied, detecting the BHB and lactate upfield resonances using a 1H homonuclear editing sequence. Plasma BHB levels also were measured. The mean brain BHB concentration increased from a nonfasted level of 0.05 +/- 0.05 to 0.60 +/- 0.26 mmol/L (after second day of fasting), increasing further to 0.98 +/- 0.16 mmol/L (after the third day of fasting). The mean nonfasted brain lactate was 0.69 +/- 0.17 mmol/L, increasing to 1.47 +/- 0.22 mmol/L after the third day. The plasma and brain BHB levels correlated well (r = 0.86) with a brain-plasma slope of 0.26. These data show that brain BHB rises significantly with 2- and 3-day fasting-induced ketosis. The lactate increase likely results from ketones displacing lactate oxidation without altering glucose phosphorylation and glycolysis.


Prevalence of acute kidney injury and prognostic significance in patients with acute myocarditis.
Myocarditis is an inflammation of the myocardium. The condition is commonly associated with rapid disease progression and often results in profound shock. Impaired renal function is the result of impairment in end-organ perfusion and is highly prevalent among critically ill patients. The aim of this study was to evaluate the incidence of acute kidney injury (AKI) and identify the relationship between AKI and the prognosis of patients with acute myocarditis. This retrospective study reviewed the medical records of 101 patients suffering from acute myocarditis between 1996 and 2011. Sixty of these patients (59%) developed AKI within 48 hours of being hospitalized. AKI defined as AKIN stage 3 (p = 0.007) and SOFA score (p = 0.03) were identified as predictors of in-hospital mortality in multivariate analysis. The conditional effect plot of the estimated risk against SOFA score upon admission categorized according to the AKIN stages showed that the risk of in-hospital mortality was highest among patients in AKIN stage 3 with a high SOFA score. Among patients with acute myocarditis, AKI defined as AKIN stage 3 and elevated SOFA score were associated with unfavorable outcomes. AKIN classification is a simple, reproducible, and easily applied evaluation tool capable of providing objective information related to the clinical prognosis of patients with acute myocarditis.


Hepatitis B and inflammatory bowel disease: role of antiviral prophylaxis.
Hepatitis B virus (HBV) is a very common infection worldwide. Its reactivation in patients receiving immunosuppression has been widely described as being associated with significant morbidity and mortality unless anti-viral prophylaxis is administered. Treatment in inflammatory bowel disease (IBD) patients has changed in recent years and immunosuppression and biological therapies are now used more frequently than before. Although current studies have reported an incidence of hepatitis B in inflammatory bowel disease patients similar to that in the general population, associated liver damage remains an important concern in this setting. Liver dysfunction may manifest in several ways, from a subtle change in serum aminotransferase levels to fulminant liver failure and death. Patients undergoing double immunosuppression are at a higher risk, and reactivation usually occurs after more than one year of treatment. As preventive measures, all IBD patients should be screened for HBV markers at diagnosis and those who are positive for the hepatitis B surface antigen should receive antiviral prophylaxis before undergoing immunosuppression in order to avoid HBV reactivation. Tenofovir/entecavir are preferred to lamivudine as nucleos(t)ide analogues due to their better resistance profile. In patients with occult or resolved HBV, viral reactivation does not appear to be a relevant issue and regular DNA determination is recommended during immunosuppression therapy. Consensus guidelines on this topic have been published in recent years. The prevention and management of HBV infection in IBD patients is addressed in this review in order to address practical recommendations.


Clinical markers of Crohn's disease severity and their association with opiate use.
The safety of opiate use for patients with Crohn's disease (CD) has long been a concern. The recent Crohn's therapy, resource, evaluation, and assessment tool (TREAT) registry update has added to these concerns by demonstrating an association of opiate use with an increased risk of infection and death in CD. While the association is clear, the relationship of opiates to these negative outcomes is not. It is unknown whether opiates are a contributing factor to these negative outcomes or if their use is merely a marker of more severe disease. We hypothesized that opiate use is not harmful in CD but is a marker of disease severity and would be associated with commonly accepted clinical markers of severe CD such as early age at CD onset, disease duration, small intestinal involvement, a history of fistula or stricture, and lower quality of life (QOL) scores. Data on CD history including pain medication usage were obtained from an interviewer directed survey of patients admitted to two tertiary care hospitals over a 2-year period. CD as the primary admitting diagnosis was not required. Active opiate use was defined by usage within the past month prior to admission. A total of 133 patients were approached to participate, of whom 108 consented to the survey, and 51 were active opiate users. Opiate using CD patients were more commonly smokers (22% vs. 3.45%, P < 0.010), had fistulas (40% vs. 22.4%, P < 0.048) and had a poorer quality of life score by short form inflammatory bowel disease questionnaire (mean 3.80 vs. 4.34, P < 0.036) than non-opiate users. No difference was found between opiate users and non-users for age of diagnosis, disease duration, or a history of strictures. The study findings demonstrate that opiate use in CD is associated with markers of disease severity including fistulas, smoking, and lower QOL scores. The findings suggest that opiates may not be directly harmful to patients with CD, but may merely be another marker of disease severity. However, given opiates unproven benefits for long term CD pain control and risk of dependence, caution should still be exercised in their use.


Peptide growth factors in the prostate as mediators of stromal epithelial interaction.
Peptide growth factors play a role in the maintenance of normal prostatic growth and differentiation (Fig. 2). It seems likely that the androgen sensitivity of human prostate is mediated by the production of peptide growth factors from stromal cells which act as the direct intermediate of androgen action on epithelial cells. TGF-beta 1 inhibition of epithelial cells is opposed by the stimulatory action of EGF, IGF and FGFs to maintain an equilibrium of epithelial cell numbers. The indirect mitogenic action of androgens appear to act by down-regulation of TGF-beta 1 and possibly EGF receptors. There is also interaction with the effects of IGF-II, produced by prostatic stromal cells and acting on epithelial cells to increase proliferation. The growth of normal prostatic fibroblasts is under the control of bFGF and TGF-beta 1. However, although our understanding of the actions of these growth factors in the normal prostate has improved over the last decade, their role in the development and maintenance of prostate cancer is less clearly defined. TGF-beta 1, classically considered to be inhibitory for epithelial cells, may be up-regulated in prostatic tumours, stimulating growth. Alternatively, autocrine production of such growth factors by tumour cells may lead to loss of inhibitory effects from exogenous TGF-beta 1, a mechanism also witnessed with TGF-alpha and bFGF. The role of EGF in the development of prostate cancer is confusing because results from the use of different cell types and experimental conditions is contradictory. It may be that a switch in the production of the predominant EGFr ligand from EGF to TGF-alpha is an important feature in the development and maintenance of the malignant phenotype. The presence of TGF-alpha autocrine loops has been shown clearly in some tumour cell lines. This switch in the production of a particular ligand may also be a feature of IGFs in prostate cancer. IGF-II may be replaced by IGF-I during malignant progression, both of which are able to act via the type 1 receptor. This change in IGF expression appears to be accompanied by altered expression of the IGF-BP2, with less detectable within prostatic tissues but elevated serum levels [58]. Basic FGF is normally produced by prostatic fibroblasts but is also produced by some prostatic cancer cell lines [64]. However, as with all growth factors, the expression of the bFGF protein and its receptor is dependent on the cell line examined. The autocrine and paracrine control of normal and abnormal prostatic growth by growth factors is important in determining their role in the development and maintenance of prostate cancer. Better understanding of such mechanisms is essential for the development of novel therapeutic strategies in the control and treatment of prostate cancer.


Theoretical investigation of the capture effect in intensity-modulation direct-detection microwave photonic links.
We introduce the capture effect concept in microwave photonic links (MWPLs) for the first time to our knowledge. The capture effect or the small-signal suppression is the change in the amplitude ratio of the two signals between input and output of the intensity-modulation direct-detection (IMDD) MWPLs. An analytical explanation of the performance of external IMDD MWPLs due to the effects of nonlinearity combined with sum of several input sinusoidal signals is given. We have investigated the suppression of a weaker signal in these links. General analytic expression for the small-signal suppression is derived using a nonlinear analytical approach. We show that the small-signal suppression is quite dependent on the input back-off, the power ratio of input signals, and on the number of input sinusoidal signals. The theoretical maximum possible signal suppression was found to be 6 dB. This analytical asymptotic value is verified by numerical results. We show the influence of the capture effect of the nonlinear MWPL on the optoelectronic oscillator operation that is verified by experimental data in the literature that has already been published.


Activation of beta-catenin signaling programs embryonic epidermis to hair follicle fate.
beta-Catenin signaling is required for hair follicle development, but it is unknown whether its activation is sufficient to globally program embryonic epidermis to hair follicle fate. To address this, we mutated endogenous epithelial beta-catenin to a dominant-active form in vivo. Hair follicle placodes were expanded and induced prematurely in activated beta-catenin mutant embryos, but failed to invaginate or form multilayered structures. Eventually, the entire epidermis adopted hair follicle fate, broadly expressing hair shaft keratins in place of epidermal stratification proteins. Mutant embryonic skin was precociously innervated, and displayed prenatal pigmentation, a phenomenon never observed in wild-type controls. Thus, beta-catenin signaling programs the epidermis towards placode and hair shaft fate at the expense of epidermal differentiation, and activates signals directing pigmentation and innervation. In transcript profiling experiments, we identified elevated expression of Sp5, a direct beta-catenin target and transcriptional repressor. We show that Sp5 normally localizes to hair follicle placodes and can suppress epidermal differentiation gene expression. We identified the pigmentation regulators Foxn1, Adamts20 and Kitl, and the neural guidance genes Sema4c, Sema3c, Unc5b and Unc5c, as potential mediators of the effects of beta-catenin signaling on pigmentation and innervation. Our data provide evidence for a new paradigm in which, in addition to promoting hair follicle placode and hair shaft fate, beta-catenin signaling actively suppresses epidermal differentiation and directs pigmentation and nerve fiber growth. Controlled downregulation of beta-catenin signaling is required for normal placode patterning within embryonic ectoderm, hair follicle downgrowth, and adoption of the full range of follicular fates.


The predicted impact of heart disease prevention and treatment initiatives on mortality in Lithuania, a middle-income country.
Disease-prevention programs compete with disease-treatment programs for scarce resources. This analysis predicts the impact of heart disease prevention and treatment initiatives for Lithuania, a middle-income Baltic country of 3.3 million people. To perform the analysis, we used data from clinical trials, the Lithuanian mortality registry, the Kaunas Monitoring of Trends and Determinants in Cardiovascular Disease (MONICA) register, Kaunas University Hospital and, when data from Lithuania were not available, the United States. We used the predicted reduction in all-cause mortality (as potentially postponable deaths) per 100,000 people aged 35 to 64 years as our outcome measure. The number of potentially postponable deaths from risk factor prevention and management in the population without apparent heart disease is 556.3 (plausible range, 282.3-878.1). The number of potentially postponable deaths for people with stable heart disease is 280.4 (plausible range, 90.8-521.8), 7.0 with a public-access defibrillator program (plausible range, 3.8-8.9), and 119.0 for hospitalized patients (plausible range, 15.9-297.7). Although improving treatment of acute events will benefit individual patients, the potential impact on the larger population is modest. Only programs that prevent and manage risk factors can generate large declines in mortality. Significant reductions in both cardiac and noncardiac death magnify the impact of risk-factor prevention and management.


Clinical guideline: management of gastroparesis.
This guideline presents recommendations for the evaluation and management of patients with gastroparesis. Gastroparesis is identified in clinical practice through the recognition of the clinical symptoms and documentation of delayed gastric emptying. Symptoms from gastroparesis include nausea, vomiting, early satiety, postprandial fullness, bloating, and upper abdominal pain. Management of gastroparesis should include assessment and correction of nutritional state, relief of symptoms, improvement of gastric emptying and, in diabetics, glycemic control. Patient nutritional state should be managed by oral dietary modifications. If oral intake is not adequate, then enteral nutrition via jejunostomy tube needs to be considered. Parenteral nutrition is rarely required when hydration and nutritional state cannot be maintained. Medical treatment entails use of prokinetic and antiemetic therapies. Current approved treatment options, including metoclopramide and gastric electrical stimulation (GES, approved on a humanitarian device exemption), do not adequately address clinical need. Antiemetics have not been specifically tested in gastroparesis, but they may relieve nausea and vomiting. Other medications aimed at symptom relief include unapproved medications or off-label indications, and include domperidone, erythromycin (primarily over a short term), and centrally acting antidepressants used as symptom modulators. GES may relieve symptoms, including weekly vomiting frequency, and the need for nutritional supplementation, based on open-label studies. Second-line approaches include venting gastrostomy or feeding jejunostomy; intrapyloric botulinum toxin injection was not effective in randomized controlled trials. Most of these treatments are based on open-label treatment trials and small numbers. Partial gastrectomy and pyloroplasty should be used rarely, only in carefully selected patients. Attention should be given to the development of new effective therapies for symptomatic control.


Design of a high-efficiency seven-port beam splitter using a dual duty cycle grating structure.
In this paper, we propose a compact seven-port beam splitter which is constructed using only a single-layer high-density grating with a dual duty cycle structure. The properties of this grating are investigated by a simplified modal method. The diffraction efficiency can be achieved around 10% more than conventional Dammann gratings while the uniformity can still be maintained at less than 1%. The effect of deviations from the design parameters on the performance of the grating is also presented.


Optical characterization of a reference instrument for gloss measurements in both a collimated and a converging beam geometry.
A reference instrument has been developed at the National Research Council of Canada for rapid, reproducible specular gloss measurements. The design and validation of this instrument for specular gloss measurements in accordance with standard methods for paints and plastics at 20 degree, 60 degree, and 85 degree geometries [American Society for Testing and Materials (ASTM) D523 and the International Organization for Standards (ISO) 2813] have been recently reported. These standard methods require a collimated beam geometry. Here we present the optical design considerations and characterization of this instrument to extend its gloss measurement capabilities to specular gloss measurements of paper samples at 75 degree geometry in accordance with standard test methods requiring a converging beam geometry (ASTM D1223 and TAPPI T480). This is, to the best of our knowledge, the first reported reference instrument that provides direct traceability for both types of standard gloss method and applications. The design challenge was to convert from a collimated beam to converging beam geometry while meeting the rigorous requirements of beam uniformity at the sample and receptor apertures specified in the 75 degree geometry test methods. We describe the innovative design to achieve this degree of functionality and reference instrument performance. The instrument's optical performance has been characterized theoretically and by comparison with measurement results. The light collection and detection systems have been analyzed via Monte Carlo simulation and ray tracing. The instrument validation includes comparison of the measurement results with theoretical gloss values for quartz, black glass, Vitrolite, and mirror gloss working standards, giving agreement of better than 0.32%. Measurement validation also involved participation in the Collaborative Testing Services program interlaboratory comparison measurements of 75 degree gloss for white papers.


Laetoli's lost tracks: 3D generated mean shape and missing footprints.
The Laetoli site (Tanzania) contains the oldest known hominin footprints, and their interpretation remains open to debate, despite over 35 years of research. The two hominin trackways present are parallel to one another, one of which is a composite formed by at least two individuals walking in single file. Most researchers have focused on the single, clearly discernible G1 trackway while the G2/3 trackway has been largely dismissed due to its composite nature. Here we report the use of a new technique that allows us to decouple the G2 and G3 tracks for the first time. In so doing we are able to quantify the mean footprint topology of the G3 trackway and render it useable for subsequent data analyses. By restoring the effectively 'lost' G3 track, we have doubled the available data on some of the rarest traces directly associated with our Pliocene ancestors.


Intravenous anti-D treatment of immune thrombocytopenic purpura: analysis of efficacy, toxicity, and mechanism of effect.
The efficacy, toxicity, and mechanism of effect of intravenous Anti-D (Winrho) were studied in 43 Rh+ patients with immune thrombocytopenia purpura (ITP) who had not undergone splenectomy and in three already splenectomized patients. The mean platelet increase for the 43 nonsplenectomized patients was 95,000/microL (median 43,000/microL). Children had greater acute platelet responses than did adults. Human immunodeficiency virus status and duration of thrombocytopenia did not affect response. Maintenance treatment was given to patients as needed: the average interval between infusions was 24 days. The three splenectomized patients had no platelet response whatsoever. Toxicity was minimal; infusions were completed in less than 5 minutes. The generally accepted mechanism of effect of Anti-D has been Fc receptor blockade by substitution of antibody-coated red blood cells for antibody-coated platelets. Evidence is presented suggesting that the effect of IV Anti-D is not limited to Fc receptor blockade, including: (1) no correlation of parameters of hemolysis with platelet increase; (2) a 48- to 72-hour delay before platelet increase; (3) a tendency of the change in monocyte Fc receptor I expression to correlate with platelet increase; and (4) increased in vitro production of antibodies to sheep red blood cells following IV Anti-D infusion.


Insula and orbital frontal cortex activity underlying emotion interference resolution in working memory.
Previous research has shown that emotional information aids conflict resolution in working memory [WM; Levens, S. M., & Phelps, E. A. Emotion processing effects on interference resolution in working memory. Journal of Emotion, 8, 267-280, 2008]. Using a recency-probes WM paradigm, it was found that positive and negative emotional stimuli reduced the amount of interference created when information that was once relevant conflicted with currently relevant information. To explore the neural mechanisms behind these facilitation effects, an event-related fMRI version of the recency-probes task was conducted using neutral and arousing positive and negative words as stimuli. Results replicate previous findings showing that the left and right inferior frontal gyrus (IFG) is involved in the interference resolution of neutral information and reveal that the IFG is involved in the interference resolution of emotional information as well. In addition, ROIs in the right and left anterior insula and in the right orbital frontal cortex (OFC) were identified that appear to underlie emotional interference resolution in WM. We conclude that the IFG underlies neutral and emotional interference resolution, and that additional regions of the anterior insula and OFC may contribute to the facilitation of interference resolution for emotional information. These findings clarify the role of the insula and OFC in affective and executive processing, specifically in WM conflict resolution.


Sumatriptan inhibits TRPV1 channels in trigeminal neurons.
To understand a possible role for transient potential receptor vanilloid 1 (TRPV1) ion channels in sumatriptan relief of pain mediated by trigeminal nociceptors. TRPV1 channels are expressed in small nociceptive sensory neurons. In dorsal root ganglia, TRPV1-containing nociceptors mediate certain types of inflammatory pain. Neurogenic inflammation of cerebral dura and blood vessels in the trigeminal nociceptive system is thought to be important in migraine pain, but the ion channels important in transducing migraine pain are not known. Sumatriptan is an agent effective in treatment of migraine and cluster headache. We hypothesized that sumatriptan might modulate activity of TRPV1 channels found in the trigeminal nociceptive system. We used immunohistochemistry to detect the presence of TRPV1 channel protein, whole-cell recording in acutely dissociated trigeminal ganglia (TG) to detect functionality of TRPV1 channels, and whole-cell recording in trigeminal nucleus caudalis (TNC) to detect effects on release of neurotransmitters from trigeminal neurons onto second order sensory neurons. Effects specifically on TG neurons that project to cerebral dura were assessed by labeling dural nociceptors with DiI. Immunohistochemistry demonstrated that TRPV1 channels are present in cerebral dura, in trigeminal ganglion, and in the TNC. Capsaicin, a TRPV1 agonist, produced depolarization and repetitive action potential firing in current clamp recordings, and large inward currents in voltage clamp recordings from acutely dissociated TG neurons, demonstrating that TRPV1 channels are functional in trigeminal neurons. Capsaicin increased spontaneous excitatory postsynaptic currents in neurons of layer II in TNC slices, showing that these channels have a physiological effect on central synaptic transmission. Sumatriptan (10 µM), a selective antimigraine drug, inhibited TRPV1-mediated inward currents in TG and capsaicin-elicited spontaneous excitatory postsynaptic currents in TNC slices. The same effects of capsaicin and sumatriptan were found in acutely dissociated DiI-labeled TG neurons innervating cerebral dura. Our results build on previous work indicating that TRPV1 channels in trigeminal nociceptors play a role in craniofacial pain. Our findings that TRPV1 is inhibited by the specific antimigraine drug sumatriptan, and that TRPV1 channels are functional in neurons projecting to cerebral dura suggests a specific role for these channels in migraine or cluster headache.


Host Genetic Control of the Oral Microbiome in Health and Disease.
Host-associated microbial communities are influenced by both host genetics and environmental factors. However, factors controlling the human oral microbiome and their impact on disease remain to be investigated. To determine the combined and relative effects of host genotype and environment on oral microbiome composition and caries phenotypes, we profiled the supragingival plaque microbiome of 485 dizygotic and monozygotic twins aged 5-11. Oral microbiome similarity always increased with shared host genotype, regardless of caries state. Additionally, although most of the variation in the oral microbiome was determined by environmental factors, highly heritable oral taxa were identified. The most heritable oral bacteria were not associated with caries state, did not tend to co-occur with other taxa, and decreased in abundance with age and sugar consumption frequency. Thus, while the human oral microbiome composition is influenced by host genetic background, potentially cariogenic taxa are likely not controlled by genetic factors.


Aquatic macrophytes in the large, sub-tropical Itaipu Reservoir, Brazil.
In the last three decades, rapid assessment surveys have become an important approach for measuring aquatic ecosystem biodiversity. These methods can be used to detect anthropogenic impacts and recognize local or global species extinctions. We present a floristic survey of the aquatic macrophytes along the Brazilian margin of the Itaipu Reservoir conducted in 2008 and compare this with a floristic survey conducted ten years earlier. We used ordination analysis to determine whether assemblage composition differed among reservoir arms. Macrophyte species were sampled in each of the 235 sampling stations using a boat, which was positioned inside three places of each macrophyte stand to record species and search for small plants. We also collected submerged plants using a rake with the boat moving at constant velocity for ten minutes. We assigned individual macrophyte species to life form and identified representative species for each life form. A total of 87 macrophyte taxa were identified. The "emergent" life forms contained the highest number of species, followed by "rooted submerged" life forms. The extensive survey of macrophytes undertaken in September 2008 recorded more species than a survey conducted between 1995 and 1998. This could be due to changes in water physico-chemistry, disturbances due to water drawdown and the long period between surveys, which may have allowed natural colonization by other species. Additionally, differences in the classification systems and taxonomic resolution used in the surveys may account for differences in the number of species recorded. Assemblage composition varied among the arms and was affected by underwater radiation (as measured using a Secchi disk) and fetch. Five non-native species were found. Two of these non-native species (Urochloa subquadripara and Hydrilla verticillata) are of special concern because they have a high frequency of occurrence and occupy large marginal areas of the reservoir. Future surveys should be conducted to determine the habitat most frequently colonized by these species. This would allow management strategies to be developed to protect native aquatic biota and prevent interference with the recreational and commercial uses of the Itaipu Reservoir.


An ITPR1 gene deletion causes spinocerebellar ataxia 15/16: a genetic, clinical and radiological description.
The purpose of this study was to characterise a novel family with very slowly progressive pure spinocerebellar ataxia (SCA) caused by a deletion in the inositol 1,4,5-triphosphate receptor 1 (ITPR1) gene on chromosome 3. This is a detailed clinical, genetic, and radiological description of the genotype. Deletions in ITPR1 have been shown to cause SCA15/SCA16 in six families to date. A further Japanese family has been identified with an ITPR1 point mutation. The exact prevalence is as yet unknown, but is probably higher than previously thought. The clinical phenotype of the family is described, and videotaped clinical examinations are presented. Serial brain magnetic resonance imaging studies were carried out on one affected individual, and genetic analysis was performed on several family members. Protein analysis confirmed the ITPR1 deletion. Affected subjects display a remarkably slow, almost pure cerebellar syndrome. Serial magnetic resonance imaging shows moderate cerebellar atrophy with mild inferior parietal and temporal cortical volume loss. Genetic analysis shows a deletion of 346,487 bp in ITPR1 (the second largest ITPR1 deletion reported to date), suggesting SCA15 is due to a loss of ITPR1 function. Western blotting of lymphoblastoid cell line protein confirms reduced ITPR1 protein levels. SCA15 is a slowly or nonprogressive pure cerebellar ataxia, which appears to be caused by a loss of ITPR1 function and a reduction in the translated protein. Patients with nonprogressive or slowly progressive ataxia should be screened for ITPR1 defects.


Acute HEV hepatitis: clinical and laboratory diagnosis.
Hepatitis E Virus (HEV) is probably the most common cause of acute hepatitis worldwide. It has been regarded for a long time as a disease limited to developing countries. Recently, the refinement of diagnostic techniques, on the one hand, and migratory flows, on the other hand, have also led to the identification of an increased number of HEV infections in industrialized countries. Four HEV genotypes have been identified across the world, with different epidemiological burdens and a wide range of clinical presentations. Here, we report a case series of acute HEV hepatitis observed in the last three years in our hospital. We performed a search for HEV IgM and IgG in all subjects admitted for acute hepatitis without evidence of other possible infectious, toxic or metabolic causes of liver damage. In subjects with HEV IgM positivity, the search for HEV-RNA was performed. We diagnosed eight acute HEV infections: 2 epidemic and 6 sporadic forms. HEV-RNA was detected in serum in 2 cases. HEV infection appears to be a cause of acute hepatitis that we must keep in mind even in developed countries.


A 7-month cigarette smoke inhalation study in C57BL/6 mice demonstrates reduced lung inflammation and emphysema following smoking cessation or aerosol exposure from a prototypic modified risk tobacco product.
Modified risk tobacco products (MRTP) are designed to reduce smoking-related health risks. A murine model of chronic obstructive pulmonary disease (COPD) was applied to investigate classical toxicology end points plus systems toxicology (transcriptomics and proteomics). C57BL/6 mice were exposed to conventional cigarette smoke (3R4F), fresh air (sham), or a prototypic MRTP (pMRTP) aerosol for up to 7 months, including a cessation group and a switching-to-pMRTP group (2 months of 3R4F exposure followed by fresh air or pMRTP for up to 5 months respectively). 3R4F smoke induced the typical adaptive changes in the airways, as well as inflammation in the lung, associated with emphysematous changes (impaired pulmonary function and alveolar damage). At nicotine-matched exposure concentrations of pMRTP aerosol, no signs of lung inflammation and emphysema were observed. Both the cessation and switching groups showed a similar reversal of inflammatory responses and no progression of initial emphysematous changes. A significant impact on biological processes, including COPD-related inflammation, apoptosis, and proliferation, was identified in 3R4F-exposed, but not in pMRTP-exposed lungs. Smoking cessation or switching reduced these perturbations to near sham-exposed levels. In conclusion, the mouse model indicated retarded disease progression upon cessation or switching to pMRTP which alone had no adverse effects.


Comparison of methods for simultaneous identification of bacterial species and determination of metabolic activity by protein-based stable isotope probing (Protein-SIP) experiments.
We developed a concept for analysing carbon and nitrogen fluxes in microbial communities by employing protein-based stable isotope probing (Protein-SIP) in metabolic labelling experiments with stable isotope labelled substrates. For identification of microbial species intact protein profiling (IPP) can be used, whereas the assessment of their metabolic activity is achieved by shotgun mass mapping (SMM). Microbial cultures were grown on substrates containing (13)C or (15)N. For identification of species we tested both the IPP and the SMM approaches. Mass spectra (MALDI-MS) were taken from mixtures of either intact proteins or peptides from tryptic digestion for generating species-specific peak patterns. In the case of SMM, the fragmentation of peptides was additionally used to obtain sequence information for species identification. Mass spectra of peptide sequences allow calculation of the amount of (13)C or (15)N incorporation within peptides for determining metabolic activity of the specific species. The comparison of IPP and SMM revealed a higher robustness of species identification by SMM. In addition, the assessment of incorporation levels of (13)C and (15)N into peptides by SMM revealed a lower uncertainty (0.5-0.8 atom %) compared to IPP (6.4-8.9 atom %). The determination of metabolic activity and function of individual species by Protein-SIP can help to analyse carbon and nitrogen fluxes within microbial communities.


Topological characterization of fractional quantum Hall ground states from microscopic Hamiltonians.
We show how to numerically calculate several quantities that characterize topological order starting from a microscopic fractional quantum Hall Hamiltonian. To find the set of degenerate ground states, we employ the infinite density matrix renormalization group method based on the matrix-product state representation of fractional quantum Hall states on an infinite cylinder. To study localized quasiparticles of a chosen topological charge, we use pairs of degenerate ground states as boundary conditions for the infinite density matrix renormalization group. We then show that the wave function obtained on the infinite cylinder geometry can be adapted to a torus of arbitrary modular parameter, which allows us to explicitly calculate the non-Abelian Berry connection associated with the modular T transformation. As a result, the quantum dimensions, topological spins, quasiparticle charges, chiral central charge, and Hall viscosity of the phase can be obtained using data contained entirely in the entanglement spectrum of an infinite cylinder.


An Overview of the Genetics of ABCA4 Retinopathies, an Evolving Story.
Stargardt disease (STGD1) and ABCA4 retinopathies (ABCA4R) are caused by pathogenic variants in the ABCA4 gene inherited in an autosomal recessive manner. The gene encodes an importer flippase protein that prevents the build-up of vitamin A derivatives that are toxic to the RPE. Diagnosing ABCA4R is complex due to its phenotypic variability and the presence of other inherited retinal dystrophy phenocopies. ABCA4 is a large gene, comprising 50 exons; to date > 2000 variants have been described. These include missense, nonsense, splicing, structural, and deep intronic variants. Missense variants account for the majority of variants in ABCA4. However, in a significant proportion of patients with an ABCA4R phenotype, a second variant in ABCA4 is not identified. This could be due to the presence of yet unknown variants, or hypomorphic alleles being incorrectly classified as benign, or the possibility that the disease is caused by a variant in another gene. This underlines the importance of accurate genetic testing. The pathogenicity of novel variants can be predicted using in silico programs, but these rely on databases that are not ethnically diverse, thus highlighting the need for studies in differing populations. Functional studies in vitro are useful towards assessing protein function but do not directly measure the flippase activity. Obtaining an accurate molecular diagnosis is becoming increasingly more important as targeted therapeutic options become available; these include pharmacological, gene-based, and cell replacement-based therapies. The aim of this review is to provide an update on the current status of genotyping in ABCA4 and the status of the therapeutic approaches being investigated.


Detection of dihydrofolate reductase conformational change by FRET using two fluorescent amino acids.
Two fluorescent amino acids, including the novel fluorescent species 4-biphenyl-l-phenylalanine (1), have been incorporated at positions 17 and 115 of dihydrofolate reductase (DHFR) to enable a study of conformational changes associated with inhibitor binding. Unlike most studies involving fluorescently labeled proteins, the fluorophores were incorporated into the amino acid side chains, and both probes [1 and L-(7-hydroxycoumarin-4-yl)ethylglycine (2)] were smaller than fluorophores typically used for such studies. The DHFR positions were chosen as potentially useful for Förster resonance energy transfer (FRET) measurements on the basis of their estimated separation (17-18 Å) and the expected change in distance along the reaction coordinate. Also of interest was the steric accessibility of the two sites: Glu17 is on the surface of DHFR, while Ile115 is within a folded region of the protein. Modified DHFR I (1 at position 17; 2 at position 115) and DHFR II (2 at position 17; 1 at position 115) were both catalytically competent. However, DHFR II containing the potentially rotatable biphenylphenylalanine moiety at sterically encumbered position 115 was significantly more active than DHFR I. Irradiation of the modified DHFRs at 280 nm effected excitation of 1, energy transfer to 2, and emission by 2 at 450 nm. However, the energy transfer was substantially more efficient in DHFR II. The effect of inhibitor binding was also measured. Trimethoprim mediated concentration-dependent diminution of the emission observed at 450 nm for DHFR II but not for DHFR I. These findings demonstrate that amino acids containing small fluorophores can be introduced into DHFR with minimal disruption of function and in a fashion that enables sensitive monitoring of changes in DHFR conformation.


A new method of intrathecal PO2, PCO2, and pH measurements for continuous monitoring of spinal cord ischemia during thoracic aortic clamping in pigs.
Impaired spinal cord circulation during thoracic aortic clamping may result in paraplegia. Reliable and fast responding methods for intraoperative monitoring are needed to facilitate the evaluation of protective measures and efficiency of revascularization. In 11 pigs, a multiparameter PO2, PCO2, and pH sensor (Paratrend 7, Biomedical Sensors Ltd, United Kingdom) was introduced into the intrathecal space for continuous monitoring of cerebrospinal fluid (CSF) oxygenation during thoracic aortic cross-clamping (AXC) distal to the left subclavian artery. A laser-Doppler probe was inserted into the epidural space for simultaneous measurements of spinal cord flux. Registrations were made before and 30 minutes after clamping and 30 and 60 minutes after declamping. The same measuring points were used for systemic hemodynamic and metabolic data acquisition. The mean CSF PO2 readings of 41 mm Hg (5.5 kPa) at baseline decreased within 3 minutes to 5 mm Hg (0.7 kPa) during AXC (P < .01). Spinal cord flux measurement responded immediately in the same way to AXC. Both methods indicated normalization of circulation during declamping. Significant (P < .01) changes were also observed in the CSF metabolic parameters PCO2 and pH. In this experimental model of spinal ischemia by AXC, online monitoring of intrathecal PO2, PCO2, and pH showed significant changes and correlated well with epidural laser-Doppler flowmetry (P < .01).


Isolation and sequence analysis of a cDNA encoding the c subunit of a vacuolar-type H(+)-ATPase from the CAM plant Kalanchoë daigremontiana.
We report the sequence of a cDNA clone encoding the c ("16 kDa') subunit of a vacuolar-type H(+)-ATPase (V-ATPase) from Kalanchoë daigremontiana, a plant in which the cell vacuole plays a pivotal role in crassulacean acid metabolism. The clone, pKVA211, was isolated from a K. daigremontiana leaf cDNA library constructed in lambda ZAP II using a homologous PCR-generated cDNA probe for the V-ATPase c subunit. The KVA211 cDNA was 839 nucleotides long and included a 20 bp poly(A)+ tail together with a complete 495 bp coding region for a polypeptide with a predicted molecular mass of 16659 Da. The deduced amino acid sequence was highly conserved across the wide range of eukaryotes (vertebrates, invertebrates, fungi, plants and protozoa) in which this gene has now been identified. Sequence comparison of several PCR products and genomic Southern analysis indicated that the V-ATPase c subunit in K. daigremontiana is encoded by a small multi-gene family. Steady-state levels of the KVA211 mRNA were much higher in leaves than in roots or flowers, and expression of this transcript in leaves was shown to be strongly light-dependent.


Mobile Block Hessian Approach with Adjoined Blocks: An Efficient Approach for the Calculation of Frequencies in Macromolecules.
In an earlier work, the authors developed a new method, the mobile block Hessian (MBH) approach, to accurately calculate vibrational modes for partially optimized molecular structures [ J. Chem. Phys. 2007 , 126 ( 22 ), 224102. ]. It is based on the introduction of blocks, consisting of groups of atoms, that can move as rigid bodies. The internal geometry of the blocks need not correspond to an overall optimization state of the total molecular structure. The standard MBH approach considers free blocks with six degrees of freedom. In the extended MBH approach introduced herein, the blocks can be connected by one or two adjoining atoms, which further reduces the number of degrees of freedom. The new approach paves the way for the normal-mode analysis of biomolecules such as proteins. It rests on the hypothesis that low-frequency modes of proteins can be described as pure rigid-body motions of blocks of consecutive amino acid residues. The method is validated for a series of small molecules and further applied to alanine dipeptide as a prototype to describe vibrational interactions between two peptide units; to crambin, a small protein with 46 amino acid residues; and to ICE/caspase-1, which contains 518 amino acid residues.


A virtual reality system for the training of volunteers involved in health emergency situations.
In order to guarantee an effective and punctual medical intervention to injured people involved in health emergency situations, where usually both professional and non-professional health operators are involved, a fast and accurate treatment has to be carried out. In case of catastrophic or very critical situations, non-professional operators who did not receive proper training (volunteers are among them) could be affected by psychological inhibitions. Their performances could slow down in such way that would affect the quality of the treatment and increase both direct and indirect costs. Our virtual reality system that is currently in use at the health care emergency center of San Martino Hospital in Genoa, Italy, has been designed and developed to check health emergency operators' capabilities to adopt correct decision-making procedures, to make optimal use of new technological equipment and to overcome psychological barriers. Our system is composed of (1) a high-end simulation PC, whose main functions are execution of the main software module, rendering of 3D scenes in stereo mode, rendering of sound, and control of data transmission from/to VR devices; (2) a low-end control PC, which controls the VR simulation running on the simulation PC, manages medical emergency simulation scenarios, introduces unexpected events to the simulation and controls the simulation difficulty level; (3) a magnetic-based motion tracking device used for head and hand tracking; (4) a wireless pair of shutter glasses together with a cathode ray tube wall projector; and (5) a high-end surround sound system. The expected benefits have been verified through the design and implementation of controlled clinical trials.


Human immunodeficiency virus type 1 pathobiology studied in humanized BALB/c-Rag2-/-gammac-/- mice.
The specificity of human immunodeficiency virus type 1 (HIV-1) for human cells precludes virus infection in most mammalian species and limits the utility of small animal models for studies of disease pathogenesis, therapy, and vaccine development. One way to overcome this limitation is by human cell xenotransplantation in immune-deficient mice. However, this has proved inadequate, as engraftment of human immune cells is limited (both functionally and quantitatively) following transplantation of mature human lymphocytes or fetal thymus/liver. To this end, a human immune system was generated from umbilical cord blood-derived CD34(+) hematopoietic stem cells in BALB/c-Rag2(-/-)gamma(c)(-/-) mice. Intrapartum busulfan administration followed by irradiation of newborn pups resulted in uniform engraftment characterized by human T-cell development in thymus, B-cell maturation in bone marrow, lymph node development, immunoglobulin M (IgM)/IgG production, and humoral immune responses following ActHIB vaccination. Infection of reconstituted mice by CCR5-coreceptor utilizing HIV-1(ADA) and subtype C 1157 viral strains elicited productive viral replication and lymphadenopathy in a dose-dependent fashion. We conclude that humanized BALB/c-Rag2(-/-)gamma(c)(-/-) mice represent a unique and valuable resource for HIV-1 pathobiology studies.


Developing object concepts in infancy: an associative learning perspective.
We present a domain-general framework called constrained attentional associative learning to provide a developmental account for how and when infants form concepts for animates and inanimates that encapsulate not only their surface appearance but also their movement characteristics. Six simulations with the same general-purpose architecture implement the features of the theory to model infant behavior in learning about objects' motion trajectory, their causal role, their onset of motion, and the initial mapping between a label and a moving object. Behavioral experiments with infants tested novel hypotheses generated by the model, showing that verbal labels initially may be associated with specific features rather than overall shape. Implications of the framework and model are discussed in relation to the mechanisms of early learning, the development of the animate-inanimate distinction, and the nature of development in the first years of life.


Supporting Self-Management of Cardiovascular Diseases Through Remote Monitoring Technologies: Metaethnography Review of Frameworks, Models, and Theories Used in Research and Development.
Electronic health (eHealth) is a rapidly evolving field informed by multiple scientific disciplines. Because of this, the use of different terms and concepts to explain the same phenomena and lack of standardization in reporting interventions often leaves a gap that hinders knowledge accumulation. Interventions focused on self-management support of cardiovascular diseases through the use of remote monitoring technologies are a cross-disciplinary area potentially affected by this gap. A review of the underlying frameworks, models, and theories that have informed projects at this crossroad could advance future research and development efforts. This research aimed to identify and compare underlying approaches that have informed interventions focused on self-management support of cardiovascular diseases through the use of remote monitoring technologies. The objective was to achieve an understanding of the distinct approaches by highlighting common or conflicting principles, guidelines, and methods. The metaethnography approach was used to review and synthesize researchers' reports on how they applied frameworks, models, and theories in their projects. Literature was systematically searched in 7 databases: Scopus, Web of Science, EMBASE, CINAHL, PsycINFO, Association for Computing Machinery Digital Library, and Cochrane Library. Included studies were thoroughly read and coded to extract data for the synthesis. Studies were mainly related by the key ingredients of the underlying approaches they applied. The key ingredients were finally translated across studies and synthesized into thematic clusters. Of 1224 initial results, 17 articles were included. The articles described research and development of 10 different projects. Frameworks, models, and theories (n=43) applied by the projects were identified. Key ingredients (n=293) of the included articles were mapped to the following themes of eHealth development: (1) it is a participatory process; (2) it creates new infrastructures for improving health care, health, and well-being; (3) it is intertwined with implementation; (4) it integrates theory, evidence, and participatory approaches for persuasive design; (5) it requires continuous evaluation cycles; (6) it targets behavior change; (7) it targets technology adoption; and (8) it targets health-related outcomes. The findings of this review support and exemplify the numerous possibilities in the use of frameworks, models, and theories to guide research and development of eHealth. Participatory, user-centered design, and integration with empirical evidence and theoretical modeling were widely identified principles in the literature. On the contrary, less attention has been given to the integration of implementation in the development process and supporting novel eHealth-based health care infrastructures. To better integrate theory and evidence, holistic approaches can combine patient-centered studies with consolidated knowledge from expert-based approaches. PROSPERO CRD42018104397; https://tinyurl.com/y8ajyajt. RR2-10.2196/13334.


Approach to buprenorphine use for opioid withdrawal treatment in the emergency setting.
Opioid use disorder (OUD) is increasing in prevalence throughout the world, with approximately three million individuals in the United States affected. Buprenorphine is a medication designed, researched, and effectively used to assist in OUD recovery. This narrative review discusses an approach to initiating buprenorphine in the emergency department (ED) for opioid-abuse recovery. Buprenorphine is a partial mu-opioid receptor agonist with high affinity and low intrinsic activity. Buprenorphine's long half-life, high potency, and 'ceiling effect' for both euphoric sensation and adverse effects make it an optimal treatment alternative for patients presenting to the ED with opioid withdrawal. While most commonly provided as a sublingual film or tablet, buprenorphine can also be delivered via transbuccal, transdermal, subdermal (implant), subcutaneous, and parenteral routes. Prior to ED administration, caution is recommended to avoid precipitation of buprenorphine-induced opioid withdrawal. Following the evaluation of common opioid withdrawal symptoms, a step-by-step approach to buprenorphine can by utilized to reach a sustained withdrawal relief. A multimodal medication-assisted treatment (MAT) plan involving pharmacologic treatment, as well as counseling and behavioral therapy, is essential to maintaining opioid remission. Patients may be safely discharged with safe-use counseling, close outpatient follow-up, and return precautions for continued management of their OUD. Establishing a buprenorphine program in the ED involves a multifactorial approach to establish a pro-buprenorphine culture. Buprenorphine is an evidence-based, safe, effective treatment option for OUD in an ED-setting. Though successfully utilized by many ED-based treatment programs, the stigma of 'replacing one opioid with another' remains a barrier. Evidence-based discussions on the safety and benefits of buprenorphine are essential to promoting a culture of acceptance and optimizing ED OUD treatment.


Health at the borders: Bayesian multilevel analysis of women's malnutrition determinants in Ethiopia.
Women's malnutrition, particularly undernutrition, remains an important public health challenge in Ethiopia. Although various studies examined the levels and determinants of women's nutritional status, the influence of living close to an international border on women's nutrition has not been investigated. Yet, Ethiopian borders are regularly affected by conflict and refugee flows, which might ultimately impact health. To investigate the impact of living close to borders in the nutritional status of women in Ethiopia, while considering other important covariates. Our analysis was based on the body mass index (BMI) of 6,334 adult women aged 20-49 years, obtained from the 2011 Ethiopian Demographic and Health Survey (EDHS). A Bayesian multilevel multinomial logistic regression analysis was used to capture the clustered structure of the data and the possible correlation that may exist within and between clusters. After controlling for potential confounders, women living close to borders (i.e. ≤100 km) in Ethiopia were 59% more likely to be underweight (posterior odds ratio [OR]=1.59; 95% credible interval [CrI]: 1.32-1.90) than their counterparts living far from the borders. This result was robust to different choices of border delineation (i.e. ≤50, ≤75, ≤125, and ≤150 km). Women from poor families, those who have no access to improved toilets, reside in lowland areas, and are Muslim, were independently associated with underweight. In contrast, more wealth, higher education, older age, access to improved toilets, being married, and living in urban or lowlands were independently associated with overweight. The problem of undernutrition among women in Ethiopia is most worrisome in the border areas. Targeted interventions to improve nutritional status in these areas, such as improved access to sanitation, economic and livelihood support, are recommended.


Functional characterization of coding polymorphisms in the human MDR1 gene using a vaccinia virus expression system.
The human MDR1-encoded transporter is a 170-kDa plasma membrane glycoprotein [P-glycoprotein (P-gp)] capable of binding and energy-dependent extrusion of structurally diverse organic compounds and drugs. P-gp seems to play a significant role in uptake, distribution, and excretion of many different drugs. To determine whether common polymorphic forms of P-gp are likely to alter function of P-gp, we characterized five known MDR1 coding polymorphisms (N21D, F103L, S400N, A893S, and A998T) using a vaccinia virus-based transient expression system. Cell surface expression of wild-type P-gp was time-dependent over a time course of 5.5 to 34.5 h; highest expression was obtained by 22 to 26.5 h after infection/transfection, indicating that a semiquantitative assay for P-gp expression levels was possible. HeLa cells stained with the P-gp specific monoclonal antibodies MRK-16 and Western blots probed with C219 revealed similar cell surface expression for the polymorphisms and for wild-type protein. Time-dependent P-gp pump function maximal at 22 h after infection/transfection was demonstrated for the following MDR1 fluorescence substrates: 4,4-difluoro-5,7-dimethyl-4-bora-3a,4a-diaza-s-indacene-3-pentanoic acid, succinimidyl ester (bodipy-FL)-verapamil, bodipy-FL-vinblastine, calcein-AM, bodipy-FL-prazosin, bisantrene, and bodipy-FL-forskolin, but not for daunorubicin. Transport studies of all tested substrates indicated that the substrate specificity of the pump was not substantially affected by any of the tested polymorphisms. Cell surface expression and function of double mutants including the more common polymorphisms (N21D-S400N, N21D-A893S, and S400N-A893S) showed no differences from wild-type. These results demonstrate that the common MDR1 coding polymorphisms result in P-gps with a cell surface distribution and function similar to wild-type P-gp.